WorldWideScience

Sample records for fusobacterium necrophorum necrobacillosis

  1. Empiema por Fusobacterium necrophorum

    Directory of Open Access Journals (Sweden)

    ML. Valle Feijoo

    2014-09-01

    Systemic infection by Fusobacerium necrophorum is characterized by the classic triad of thrombosis of the jugular internal vein, bacteriemia and metastatic spread and it is known as Lemiere syndrome (LS, post-amigdalitis sepsis or necrobacillosis. Pulmonary impairement due to septic embolism is very common in LS, but Fusobacterium necrophorum is hardly ever found in empyemas without LS. We report a new case.

  2. Characterization of Fusobacterium necrophorum isolated from llama and alpaca.

    Science.gov (United States)

    Kumar, Amit; Anderson, David; Amachawadi, Raghavendra G; Nagaraja, Tiruvoor G; Narayanan, Sanjeev K

    2013-07-01

    Fusobacterium necrophorum, a Gram-negative, anaerobic bacterium, is an opportunistic animal and human pathogen that causes a variety of infections termed necrobacillosis. There are 2 subspecies of F. necrophorum (subsp. necrophorum and subsp. funduliforme) that differ morphologically and biochemically and in virulence. Leukotoxin, a secreted protein, is considered to be the major virulence factor. In camelids, F. necrophorum causes a variety of infections, generally involving the lips, tongue, pharynx, interdigital spaces, foot pad, larynx, mandible, or maxillary bones. The objective of the current study was to characterize the presumptive Fusobacterium isolates from a variety of necrotic infections in llama (Lama glama) and alpaca (Vicugna pacos) and determine whether the strains possess leukotoxin activities. A total of 7 isolates from alpaca and 2 isolates from llama were characterized. Based on growth characteristics in broth culture, and biochemical and polymerase chain reaction analyses, all 9 isolates belonged to subsp. necrophorum and possessed the putative hemagglutinin gene. Western blot analysis with antileukotoxin antibodies raised in rabbit showed the presence of leukotoxin protein in the culture supernatant of all isolates. Furthermore, flow cytometry of the culture supernatants demonstrated cytotoxicity to bovine and alpaca polymorphonuclear leukocytes (PMNs). The extent of cytotoxicity to either alpaca or bovine PMNs differed among camelid strains. The cytotoxicity of many of the camelid strains was higher (P llama and alpaca are similar to bovine isolates, and leukotoxin may be a major virulence factor.

  3. Fusobacterium necrophorum presenting as isolated lung nodules

    Directory of Open Access Journals (Sweden)

    Rajiv Sonti

    2015-01-01

    Full Text Available Fusobacterium necrophorum causes Lemierre's syndrome - a dramatic and distinct condition beginning with pharyngitis before proceeding to internal jugular vein septic thrombophlebitis and respiratory tract infection in otherwise healthy individuals. It is rare, but by far the most common pathway to parenchymal lung disease with this organism. Here we describe we a 34 year old healthy lady who was nontoxic without any antecedent illness who presented with lung nodules due to fusobacterium necrophorum as the sole manifestation of disease. Leading diagnostic consideration prior to culture data was pulmonary vasculitis. Identifying her disease process was a somewhat chance occurrence, and it began to resolve prior to antibiotic therapy. Though it would be difficult to recommend keen awareness of this organism given its rarity, it is important to consider that its scope may be broader than traditionally considered.

  4. Fusobacterium necrophorum in North American Bighorn Sheep ( Ovis canadensis ) Pneumonia.

    Science.gov (United States)

    Shanthalingam, Sudarvili; Narayanan, Sanjeevkumar; Batra, Sai Arun; Jegarubee, Bavananthasivam; Srikumaran, Subramaniam

    2016-07-01

    Fusobacterium necrophorum has been detected in pneumonic bighorn sheep (BHS; Ovis canadensis ) lungs, in addition to the aerobic respiratory pathogens Mannheimia haemolytica , Bibersteinia trehalosi , Pasteurella multocida , and Mycoplasma ovipneumoniae . Similar to M. haemolytica , F. necrophorum produces a leukotoxin. Leukotoxin-induced lysis and degranulation of polymorphonuclear leukocytes (PMNs) and macrophages are responsible for acute inflammation and lung tissue damage characteristic of M. haemolytica -caused pneumonia. As one approach in elucidating the role of F. necrophorum in BHS pneumonia, we determined the frequency of the presence of F. necrophorum in archived pneumonic BHS lung tissues, and susceptibility of BHS leukocytes to F. necrophorum leukotoxin. A species-specific PCR assay detected F. necrophorum in 37% of pneumonic BHS lung tissues (total tested n=70). Sequences of PCR amplicons were similar to the less virulent F. necrophorum subsp. funduliforme. Fusobacterium necrophorum leukotoxin exhibited cytotoxicity to BHS PMNs and peripheral blood mononuclear cells. As with the M. haemolytica leukotoxin, F. necrophorum leukotoxin was more toxic to BHS PMNs than domestic sheep PMNs. It is likely that F. necrophorum enters the lungs after M. haemolytica and other aerobic respiratory pathogens enter the lungs and initiate tissue damage, thereby creating a microenvironment that is conducive for anaerobic bacterial growth. In summary, Fusobacterium leukotoxin is highly toxic for BHS leukocytes; however, based on the PCR findings, it is unlikely to play a direct role in the development of BHS pneumonia.

  5. Livstruende appendicitis forårsaget af Fusobacterium necrophorum

    DEFF Research Database (Denmark)

    Hagen, Trine Langfeldt; Maeda, Yasuko; Lindberg, Jens Aage

    2014-01-01

    Fusobacterium necrophorum is a well-known cause of Lemierre's syndrome. Recent studies suggest a causative association between F. necrophorum and acute appendicitis. We present a case of a 15-year-old previously healthy girl who presented with acute non-perforated appendicitis, intra......-abdominal abscesses and thrombosis that led to omental necrosis. This resulted in a life-threatening septic shock with the need for prolonged intensive care. We suggest that F. necrophorum identified in pus from the abdomen caused this fulminant variation of appendicitis with findings similar to those seen...

  6. Necrotizing pneumonia after pharyngitis due to fusobacterium necrophorum

    Energy Technology Data Exchange (ETDEWEB)

    Kleinman, P K; Flowers, R A

    1984-01-01

    A case of necrotizing pneumonia secordary to Fusobacterium necrophorum is reported. This anaerobic infection commonly originates in the upper respiratory tract and is often accompanied by multiple system disease due to hematogeneous seeding. When the lungs are involved, diffuse necrotizing pneumonia with pleural effusions and cavitation result. The course is prolonged, and the diagnosis is frequently delayed. With appropriate antibiotics, the prognosis is good.

  7. Necrobacillosis - A resurgence?

    International Nuclear Information System (INIS)

    Liu, Andrea C.Y.; Argent, Julie D.

    2002-01-01

    Necrobacillosis is a rare life threatening illness caused by Fusobacterium necrophorum. It usually affects previously healthy adolescents and presents as symptomatic pharyngeal infection followed by bacteraemia and metastatic abscesses. A high degree of clinical suspicion is needed because there is often a delay between the presentation of sore throat and the development of systemic illness. The clinical and radiological features of four cases of necrobacillosis are reported and a review of the spectrum of manifestations of metastatic infection presented. Liu, A.C.Y. and Argent, J.D. (2002)

  8. Liver abscess caused by periodontal bacterial infection with Fusobacterium necrophorum.

    Science.gov (United States)

    Yoneda, Masato; Kato, Shingo; Mawatari, Hironori; Kirikoshi, Hiroyuki; Imajo, Kento; Fujita, Koji; Endo, Hiroki; Takahashi, Hirokazu; Inamori, Masahiko; Kobayashi, Noritoshi; Kubota, Kensuke; Saito, Satoru; Tohnai, Iwai; Watanuki, Kei; Wada, Koichiro; Maeda, Shin; Nakajima, Atsushi

    2011-02-01

    Liver abscess is recognized as a life-threatening disease. However, even in recent years, approximately 50% of liver abscess cases are considered to be cryptogenic. Here, we report a case of liver abscess associated with periodontal bacterial infection by Fusobacterium necrophorum, which is commonly found in the oropharyngeal flora. A 36-year-old man presented with fever and contrast-enhanced abdominal computed tomography revealed multiple liver abscesses. F.necrophorum was isolated from oral smears, liver aspirates and blood samples. Liver abscesses caused by periodontal bacterial infection are rare, however, the incidence is expected to increase in the future, as periodontitis is extremely common and is on the rise as one of the most common chronic infections in the world. A systemic survey including periodontitis may be required for the exact diagnosis of the source of infection. © 2011 The Japan Society of Hepatology.

  9. Ribotyping to compare Fusobacterium necrophorum isolates from bovine liver abscesses, ruminal walls, and ruminal contents.

    OpenAIRE

    Narayanan, S; Nagaraja, T G; Okwumabua, O; Staats, J; Chengappa, M M; Oberst, R D

    1997-01-01

    Restriction fragment length polymorphism analysis of rRNA genes was employed to genetically compare Fusobacterium necrophorum subsp. necrophorum and F. necrophorum subsp. funduliforme isolates from multiple abscesses of the same liver and isolates from liver abscesses, the ruminal wall, and ruminal contents from the same animal. Four livers with multiple abscesses and samples of ruminal contents, ruminal walls, and liver abscesses were collected from 11 cattle at slaughter. F. necrophorum was...

  10. Fusobacterium necrophorum otitis and mastoiditis in infants and young toddlers.

    Science.gov (United States)

    Stergiopoulou, T; Walsh, T J

    2016-05-01

    There is an increased recovery of Fusobacterium necrophorum from cases of otitis media and mastoiditis in the pediatric population. These infections may be highly severe, causing local osteomyelitis, bacteremia, and Lemierre's syndrome. The severity and difficulties in providing optimal treatment for these infections may be especially difficult in this age group due to immunological immaturity and delayed presentation. In this review of literature, we present and analyze the clinical presentation, management, and outcome of otic infections caused by F. necrophorum in infants and young toddlers less than 2 years old. Search in Pubmed was conducted for reported cases in the English literature for the time period of the last 50 years. Twelve well-described cases were retrieved with F. necrophorum otitis and mastoiditis and complications reported in all cases. Treatment included both intravenously with antimicrobial agents (beta lactams plus metronidazole) and mastoidectomy. Lemierre's syndrome and Lemierre's syndrome variants developed in 60 % of the patients. Dissemination of the infection as distal osteomyelitis and septic shock were also reported. The outcome was favorable in all the cases. Otitis and mastoiditis infections in children less then 2 years old are invasive infections, and severe complications can occur.

  11. Detection of Fusobacterium necrophorum subsp funduliforme in tonsillitis in young adults by real-time PCR

    DEFF Research Database (Denmark)

    Jensen, Anders; Hagelskjær Kristensen, Lena; Prag, Jørgen

    2007-01-01

    Throat swabs from 61 patients, aged 18-32 years, with non-streptococcal tonsillitis (NST) and 92 healthy controls were examined for the presence of Fusobacterium necrophorum DNA using a novel TaqMan-based real-time quantitative PCR assay for F. necrophorum subspecies. The assay was based on the g...

  12. Fusobacterium necrophorum Pharyngitis Complicated by Lemierre’s Syndrome

    Directory of Open Access Journals (Sweden)

    Antonio Faraone

    2016-01-01

    Full Text Available We report the case of an 18-year-old woman who was referred to our outpatient clinic because of a 2-week history of sore throat, high fever, and neck tenderness unresponsive to a 7-day amoxicillin/clavulanic acid course. Infectious mononucleosis was initially suspected, but an extremely high value of procalcitonin and clinical deterioration suggested a bacterial sepsis, prompting the patient admission to our internal medicine ward. Blood cultures were positive for Fusobacterium necrophorum. CT scan detected a parapharyngeal abscess, a right internal jugular vein thrombosis, and multiple bilateral lung abscesses, suggesting the diagnosis of Lemierre’s syndrome. The patient was treated with a 2-week course of metronidazole and meropenem with a gradual clinical recovery. She was thereafter discharged home with metronidazole and amoxicillin/clavulanic acid for 14 days and a 3-month course of enoxaparin, experiencing an uneventful recovery. The present case highlights the importance of taking into consideration the Lemierre’s syndrome whenever a pharyngotonsillitis has a severe and unusual course.

  13. Frequency, microbial interactions, and antimicrobial susceptibility of Fusobacterium nucleatum and Fusobacterium necrophorum isolated from primary endodontic infections.

    Science.gov (United States)

    Jacinto, Rogério C; Montagner, Francisco; Signoretti, Fernanda G C; Almeida, Geovania C; Gomes, Brenda P F A

    2008-12-01

    This study assessed the prevalence and microbial interactions of Fusobacterium nucleatum and Fusobacterium necrophorum in primary endodontic infections from a Brazilian population and their antimicrobial susceptibility to some antibiotics by the E-test. One hundred ten samples from infected teeth with periapical pathologies were analyzed by culture methods. Five hundred eighty individual strains were isolated; 81.4% were strict anaerobes. F. nucleatum was found in 38 root canals and was associated with Porphyromonas gingivalis, Prevotella spp., and Eubacterium spp. F. necrophorum was found in 20 root canals and was associated with Peptostreptococcus prevotii. The simultaneous presence of F. nucleatum and F. necrophorum was not related to endodontic symptoms (p > 0.05). They were 100% susceptible to amoxicillin, amoxicillin/clavulanate, and cephaclor. Fusobacterium spp. is frequently isolated from primary-infected root canals of teeth with periapical pathologies. Amoxicillin is a useful antibiotic against F. nucleatum and F. necrophorum in endodontic infections and has been prescribed as the first choice in Brazil.

  14. Human Infection with Fusobacterium necrophorum without Jugular Venous Thrombosis: A Varied Presentation of Lemierre’s Syndrome

    Directory of Open Access Journals (Sweden)

    Muhammad Asim Rana

    2017-01-01

    Full Text Available Lemierre’s syndrome is also known as postangina septicemia, which is commonly caused by Fusobacterium necrophorum also known as Necrobacillus and also by other microorganisms like Staphylococcus, Streptococcus, Peptostreptococcus, and Bacteroides. Though the disease starts as an upper respiratory tract infection, it may spread and cause thrombophlebitis of the internal jugular vein. It may present itself through cranial nerve palsy or sepsis involving distant organs like the lungs or bones. It is also known as forgotten disease because of its rarity. Fusobacterium necrophorum usually causes infection in animals and rarely affects humans. We hereby present a case of Necrobacillus infection which did not cause any thrombophlebitis but resulted in severe pneumonia and acute kidney injury, leading to respiratory failure and requiring mechanical ventilation.

  15. Hemolytic activity of Fusobacterium necrophorum culture supernatants due to presence of phospholipase A and lysophospholipase.

    Science.gov (United States)

    Abe, P M; Kendall, C J; Stauffer, L R; Holland, J W

    1979-01-01

    Culture supernatants of Fusobacterium necrophorum demonstrated hemolytic activity. The hemolysin(s), which was partially purified by ammonium sulfate precipitation, was temperature-dependent and heat labile. The spectrum of hemolytic activity against various erythrocytes included rabbit, human, and dog erythrocytes. Goats, sheep, and bovine erythrocytes showed only trace hemolysis. According to results of thin-layer chromatography, the hemolysin hydrolyzed rabbit erythrocyte phosphatidyl choline, phosphatidyl ethanolamine, lysophosphatidyl choline, and bovine phosphatidyl choline. Hydrolysis of egg yolk phosphatidyl choline, bovine phosphatidyl ethanolamine, cholesterol, 1,2-dipalmitin, 1,3-dipalmitin, sphingomyelin, or triolein was not detected by thin layer chromatography. A more sensitive procedure utilizing gas-liquid chromatography revealed that, of the substrates tested, the following were bein hydrolyzed: bovine and egg yolk phosphatidyl choline, lysophosphatidyl choline, alpha-palmito-beta-eleoyl-L-alpha lecithin and alpha-oleoyl-betal-palmitoyl-L-alpha lecithin. Substrates which were weakly hydrolyzed were bovine phosphatidyl ethanolamine, DL-alpha-hosphatidyl ethanolamine dipalmitoyl, 1,2-dipalmitin, 1,3-dipalmitin, and triolein.

  16. Mathematical modeling of ovine footrot in the UK: the effect of Dichelobacter nodosus and Fusobacterium necrophorum on the disease dynamics

    Directory of Open Access Journals (Sweden)

    Jolene Atia

    2017-12-01

    Full Text Available Dichelobacter nodosus is a virulent, invasive, anaerobic bacterium that is believed to be the causative agent of ovine footrot, an infectious bacterial disease of sheep that causes lameness. Another anaerobe, Fusobacterium necrophorum, has been intimately linked with the disease occurrence and severity. Here we examine data from a longitudinal study of footrot on one UK farm, including quantitative PCR (qPCR estimates of bacterial load of D. nodosus and F. necrophorum. The data is at foot level; all feet were monitored for five weeks assessing disease severity (healthy, interdigital dermatitis (ID, or severe footrot (SFR and bacterial load (number of bacteria/swab. We investigate the role of D. nodosus and F. necrophorum in the progress of the disease using a continuous-time Markov model with 12 different states characterising the foot. The transition rates between the adjacent states are the (34 model parameters, these are determined using Metropolis Hasting MCMC. Our aim is to determine the predictive relationship between past and future D. nodosus and F. necrophorum load and disease states. We demonstrate a high level of predictive accuracy at the population level for the D. nodosus model, although the dynamics of individual feet is highly stochastic. However, we note that this predictive accuracy at population level is only high in more diseased states for F. necrophorum model. This supports our hypothesis that D. nodosus load and status of the foot work in combination to give rise to severe footrot and lameness, and that D. nodosus load plays the primary role in the initiation and progression of footrot, while F. necrophorum load rather increases disease severity of SFR.

  17. Mixed species biofilms of Fusobacterium necrophorum and Porphyromonas levii impair the oxidative response of bovine neutrophils in vitro.

    Science.gov (United States)

    Lockhart, Joey S; Buret, Andre G; Ceri, Howard; Storey, Douglas G; Anderson, Stefanie J; Morck, Douglas W

    2017-10-01

    Biofilms composed of anaerobic bacteria can result in persistent infections and chronic inflammation. Host immune cells have difficulties clearing biofilm-related infections and this can result in tissue damage. Neutrophils are a vital component of the innate immune system and help clear biofilms. The comparative neutrophilic response to biofilms versus planktonic bacteria remains incompletely understood, particularly in the context of mixed infections. The objective of this study was to generate mixed species anaerobic bacterial biofilms composed of two opportunistic pathogens, Fusobacterium necrophorum and Porphyromonas levii, and evaluate neutrophil responses to extracellular fractions from both biofilms and planktonic cell co-cultures of the same bacteria. Purified bovine neutrophils exposed to culture supernatants from mixed species planktonic bacteria showed elevated oxidative activity compared to neutrophils exposed to biofilms composed of the same bacteria. Bacterial lipopolysaccharide plays a significant role in the stimulation of neutrophils; biofilms produced substantially more lipopolysaccharide than planktonic bacteria under these experimental conditions. Removal of lipopolysaccharide significantly reduced neutrophil oxidative response to culture supernatants of planktonic bacteria. Oxidative responses to LPS-removed biofilm supernatants and LPS-removed planktonic cell supernatants were similar. The limited neutrophil response to biofilm bacteria observed in this study supports the reduced ability of the innate immune system to eradicate biofilm-associated infections. Lipopolysaccharide is likely important in neutrophil response; however, the presence of other extracellular, immune modifying molecules in the bacterial media also appears to be important in altering neutrophil function. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Subcutaneous immunization with inactivated bacterial components and purified protein of Escherichia coli, Fusobacterium necrophorum and Trueperella pyogenes prevents puerperal metritis in Holstein dairy cows.

    Science.gov (United States)

    Machado, Vinícius Silva; Bicalho, Marcela Luccas de Souza; Meira Junior, Enoch Brandão de Souza; Rossi, Rodolfo; Ribeiro, Bruno Leonardo; Lima, Svetlana; Santos, Thiago; Kussler, Arieli; Foditsch, Carla; Ganda, Erika Korzune; Oikonomou, Georgios; Cheong, Soon Hon; Gilbert, Robert Owen; Bicalho, Rodrigo Carvalho

    2014-01-01

    In this study we evaluate the efficacy of five vaccine formulations containing different combinations of proteins (FimH; leukotoxin, LKT; and pyolysin, PLO) and/or inactivated whole cells (Escherichia coli, Fusobacterium necrophorum, and Trueperella pyogenes) in preventing postpartum uterine diseases. Inactivated whole cells were produced using two genetically distinct strains of each bacterial species (E. coli, F. necrophorum, and T. pyogenes). FimH and PLO subunits were produced using recombinant protein expression, and LKT was recovered from culturing a wild F. necrophorum strain. Three subcutaneous vaccines were formulated: Vaccine 1 was composed of inactivated bacterial whole cells and proteins; Vaccine 2 was composed of proteins only; and Vaccine 3 was composed of inactivated bacterial whole cells only. Two intravaginal vaccines were formulated: Vaccine 4 was composed of inactivated bacterial whole cells and proteins; and Vaccine 5 was composed of PLO and LKT. To evaluate vaccine efficacy, a randomized clinical trial was conducted at a commercial dairy farm; 371 spring heifers were allocated randomly into one of six different treatments groups: control, Vaccine 1, Vaccine 2, Vaccine 3, Vaccine 4 and Vaccine 5. Late pregnant heifers assigned to one of the vaccine groups were each vaccinated twice: at 230 and 260 days of pregnancy. When vaccines were evaluated grouped as subcutaneous and intravaginal, the subcutaneous ones were found to significantly reduce the incidence of puerperal metritis. Additionally, subcutaneous vaccination significantly reduced rectal temperature at 6±1 days in milk. Reproduction was improved for cows that received subcutaneous vaccines. In general, vaccination induced a significant increase in serum IgG titers against all antigens, with subcutaneous vaccination again being more effective. In conclusion, subcutaneous vaccination with inactivated bacterial components and/or protein subunits of E. coli, F. necrophorum and T. pyogenes

  19. Digital Necrobacillosis in Norwegian Wild Tundra Reindeer (Rangifer tarandus tarandus)

    DEFF Research Database (Denmark)

    Handeland, K.; Boye, Mette; Bergsjø, B.

    2010-01-01

    Outbreaks of digital necrobacillosis in Norwegian wild tundra reindeer (Rangifer tarandus tarandus) are described. The outbreaks occurred in late summer and autumn 2007 and 2008, subsequent to periods with an unusually high number of days with precipitation and high air temperature. Lesions were....... necrophorum was cultured from the foot lesions of six animals. Five of these isolates were examined by 16S rRNA sequencing. The sequences were identical and differed from all other strains listed in GenBank. These results are consistent with circulation of a reindeer-adapted pathogenic strain of F....... necrophorum in the wild reindeer population, causing outbreaks of digital necrobacillosis following warm and humid summers....

  20. Prevotella bivia necrobacillosis following infectious mononucleosis

    NARCIS (Netherlands)

    Huits, R.M.H.G.; van Assen, S.; Wildeboer- Veloo, Alida; Verschuuren, E.A.M.; Koeter, G.H.

    2006-01-01

    A case of Lemierre's syndrome is reported. Although Fusobacterium species are commonly associated with this presentation, Prevotella bivia was the causative micro-organism identified in this case. The finding that disseminated anaerobic sepsis followed primary EBV infection led to the construction

  1. Term stillbirth caused by oral Fusobacterium nucleatum.

    Science.gov (United States)

    Han, Yiping W; Fardini, Yann; Chen, Casey; Iacampo, Karla G; Peraino, Victoria A; Shamonki, Jaime M; Redline, Raymond W

    2010-02-01

    Intrauterine infection is a recognized cause of adverse pregnancy outcome, but the source of infection is often undetermined. We report a case of stillbirth caused by Fusobacterium nucleatum that originated in the mother's mouth. A woman with pregnancy-associated gingivitis experienced an upper respiratory tract infection at term, followed by stillbirth a few days later. F. nucleatum was isolated from the placenta and the fetus. Examination of different microbial floras from the mother identified the same clone in her subgingival plaque but not in the supragingival plaque, vagina, or rectum. F. nucleatum may have translocated from the mother's mouth to the uterus when the immune system was weakened during the respiratory infection. This case sheds light on patient management for those with pregnancy-associated gingivitis.

  2. Fusobacterium nucleatum in endodontic flare-ups.

    Science.gov (United States)

    Chávez de Paz Villanueva, Luis Eduardo

    2002-02-01

    The extent to which Fusobacterium nucleatum is recovered from root canals of teeth that present with an interappointment flare-up following endodontic instrumentation was investigated. Included in the study were 28 patients that sought emergency treatment after initiation of root canal therapy. Only non-painful teeth that had been treated because of a necrotic pulp and periapical inflammatory lesion were studied. Root canal samples for bacterial analysis were taken, transported to a bacteriological laboratory, and processed for a semiquantitative assessment of bacterial isolates. Bacterial findings were correlated with self-assessed pain intensity as recorded by means of a Visual Analogue Scale. Clinical presentation of swelling and presence of exudate in the treated root canals were also linked. Bacteria were recovered from all teeth examined. Gram-negative anaerobic coccoid rods (Prevotella species and Porphyromonas species) were frequent isolates. All teeth in patients who were reported to be in severe pain (Visual Analogue Scale > or = 6) displayed F nucleatum. Nine out of 10 of these teeth also had swelling and exudate in the root canals. Samples from the remaining patients that had teeth with less pain score showed a variable bacterial recovery. None of these teeth displayed F nucleatum. F nucleatum appears to be associated with the development of the most severe forms of interappointment endodontic flare-ups.

  3. Myopericarditis associated with Fusobacterium nucleatum-caused liver abscess.

    Science.gov (United States)

    Kearney, Alexis; Knoll, Bettina

    2015-03-01

    A wide clinical spectrum of bacteremic disease caused by Fusobacterium has been presented in this journal. We wish to extend this spectrum by presenting a case of myopericarditis resulting from a liver abscess caused by F. nucleatum. While F. nucleatum plays an important role in periodontal disease, and has been isolated from skin ulcers, liver abscesses, urinary tract infections, and endocarditis, a single case of F. nucleatum-induced pericarditis is documented in the literature.

  4. Ovine fetal necrobacillosis

    DEFF Research Database (Denmark)

    Agerholm, J.S.; Boye, Mette; Aalbæk, B.

    2007-01-01

    were found in several tissues. Histologically, placental lesions were characterized by locally diffuse infiltration of neutrophils, closely associated with abundant small Gram-negative and FISH-positive rods, thrombosis and necrosis. Lesions in the fetal-maternal interface were multifocal and consisted...

  5. Clinical Features and Outcomes of Fusobacterium Species Infections in a Ten-Year Follow-up

    Directory of Open Access Journals (Sweden)

    Garcia-Carretero Rafael

    2017-10-01

    Full Text Available Objective: Although uncommon, Fusobacterium infections have a wide clinical spectrum, ranging from local pharyngeal infections to septic shock. Our aim was to characterize and analyze the clinical features and outcomes in patients with Fusobacterium infections, and determine which variables were able to predict a poor outcome.

  6. Synergy in biofilm formation between Fusobacterium nucleatum and Prevotella species.

    Science.gov (United States)

    Okuda, Tamaki; Kokubu, Eitoyo; Kawana, Tomoko; Saito, Atsushi; Okuda, Katsuji; Ishihara, Kazuyuki

    2012-02-01

    The formation of biofilm by anaerobic, Gram-negative bacteria in the subgingival crevice plays an important role in the development of chronic periodontitis. The aim of this study was to characterize the role of coaggregation between Fusobacterium nucleatum and Prevotella species in biofilm formation. Coaggregation between F. nucleatum and Prevotella species was determined by visual assay. Effect of co-culture of the species on biofilm formation was assessed by crystal violet staining. Effect of soluble factor on biofilm formation was also examined using culture supernatant and two-compartment co-culture separated by a porous membrane. Production of autoinducer-2 (AI-2) by the organisms was evaluated using Vibrio harveyi BB170. Cells of all F. nucleatum strains coaggregated with Prevotella intermedia or Prevotella nigrescens with a score of 1-4. Addition of ethylenediamine tetraacetic acid or l-lysine inhibited coaggregation. Coaggregation disappeared after heating of P. intermedia or P. nigrescens cells, or Proteinase K treatment of P. nigrescens cells. Co-culture of F. nucleatum ATCC 25586 with P. intermedia or P. nigrescens strains increased biofilm formation compared with single culture (p culture with culture supernatant of these strains, however, did not enhance biofilm formation by F. nucleatum. Production of AI-2 in Prevotella species was not related to enhancement of biofilm formation by F. nucleatum. These findings indicate that physical contact by coaggregation of F. nucleatum strains with P. intermedia or P. nigrescens plays a key role in the formation of biofilm by these strains. Copyright © 2011 Elsevier Ltd. All rights reserved.

  7. Noncontiguous finished genome sequence and description of Fusobacterium massiliense sp. nov. isolated from human duodenum

    Directory of Open Access Journals (Sweden)

    M. Mailhe

    2017-03-01

    Full Text Available The strain Marseille-P2749T (= CSUR P2749=DSM 103085 was isolated as part of culturomics study from a liquid duodenum sample from a French man. Bacterial cells were Gram-negative bacilli, fusiform shaped and non–spore forming, and they grew in microaerophilic and anaerobic atmosphere. Its genome is 1 809 169 bp long and contains 1646 protein-coding genes. The DNA G+C content was 27.33 mol%. This strain exhibited a 95.9% sequence similarity with Fusobacterium periodonticum, the phylogenetically closest species with standing in nomenclature. Strain Marseille-P2749T is suggested to be a novel species belonging to the genus Fusobacterium, for which the name Fusobacterium massiliense sp. nov. is proposed.

  8. 5S ribosomal ribonucleic acid sequences in Bacteroides and Fusobacterium: evolutionary relationships within these genera and among eubacteria in general

    Science.gov (United States)

    Van den Eynde, H.; De Baere, R.; Shah, H. N.; Gharbia, S. E.; Fox, G. E.; Michalik, J.; Van de Peer, Y.; De Wachter, R.

    1989-01-01

    The 5S ribosomal ribonucleic acid (rRNA) sequences were determined for Bacteroides fragilis, Bacteroides thetaiotaomicron, Bacteroides capillosus, Bacteroides veroralis, Porphyromonas gingivalis, Anaerorhabdus furcosus, Fusobacterium nucleatum, Fusobacterium mortiferum, and Fusobacterium varium. A dendrogram constructed by a clustering algorithm from these sequences, which were aligned with all other hitherto known eubacterial 5S rRNA sequences, showed differences as well as similarities with respect to results derived from 16S rRNA analyses. In the 5S rRNA dendrogram, Bacteroides clustered together with Cytophaga and Fusobacterium, as in 16S rRNA analyses. Intraphylum relationships deduced from 5S rRNAs suggested that Bacteroides is specifically related to Cytophaga rather than to Fusobacterium, as was suggested by 16S rRNA analyses. Previous taxonomic considerations concerning the genus Bacteroides, based on biochemical and physiological data, were confirmed by the 5S rRNA sequence analysis.

  9. Decreased interleukin-2 responses to Fusobacterium nucleatum and Porphyromonas gingivalis in generalized aggressive periodontitis

    DEFF Research Database (Denmark)

    Borch, Tanja Skuldbøl; Løbner, Morten; Bendtzen, Klaus

    2009-01-01

    with disease-relevant pathogens. METHODS: Mononuclear cells (MNCs) from 10 white patients with GAgP and 10 white controls were stimulated with Porphyromonas gingivalis American Type Culture Collection (ATCC) 33277 (Pg), Prevotella intermedia ATCC 25611, Fusobacterium nucleatum ATCC 49256 (Fn), and similar...

  10. Occurrence and antimicrobial susceptibility of Porphyromonas spp. and Fusobacterium spp. in dogs with and without periodontitis.

    Science.gov (United States)

    Senhorinho, Gerusa N A; Nakano, Viviane; Liu, Chengxu; Song, Yuli; Finegold, Sydney M; Avila-Campos, Mario J

    2012-08-01

    The occurrence of Porphyromonas gulae, Porphyromonas macacae, Fusobacterium nucleatum and Fusobacterium canifelinum in subgingival plaque from dogs with and without periodontitis as well as their antimicrobial susceptibility were evaluated. From 50 dogs with periodontitis were identified 38 P. gulae, 8 P. macacae, 26 F. nucleatum and 15 F. canifelinum, and from 50 dogs without periodontitis were identified 15 P. gulae, 12 F. nucleatum and 11 F. canifelinum. All strains were susceptible to most of the antibiotics tested, however, different resistance rates to clarithromycin, erythromycin and metronidazole among strains were observed. The role of P. gulae, P. macacae, F. nucleatum and F. canifelinum in periodontal disease of household pets needs to be defined to a better prevention and treatment of the canine periodontitis. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. Rendimento dos meios seletivos para bactérias do gênero Fusobacterium

    Directory of Open Access Journals (Sweden)

    Wilson C. de Araújo

    1968-10-01

    Full Text Available Meios seletivos para Fusobacterium foram, comparados no aspecto de sua eficiência no isolamento dos microrganismos fusiformes e quanto à inibição da flora concomitante. Material não calcificado do sulco gengival foi homogeneizado e diluído em tampão de fosfato (0,067 M - pH 7,2; 0,1 ml foi semeado em agar sangue, no meio de Omata & Disraely, meio de Omata & Disraely sem sôro, meio de McMarthy & Snyder, agar sangue com vancomicina, meio de Onisi, meio de Onisi com sôro de cavalo e meio de Onisi com pH 7,0. Em todos os casos a incubação foi realizada em anaerobiose, em jarra tipo Brewer, com 95% nitrogênio e 5% gás carbônico, durante 4 dias. O meio de McCarthy & Snyder mostrou-se tão eficiente quanto o agar sangue em relação ao isolamento de Fusobacterium, ainda inibindo a flora concomitante, o meio de Omata & Disraely, com cristal violeta e estreptomicina, foi de certo modo inibidor também para Fusobacterium. O meio de Onisi, sugerido, sem sôro, possibilitou o isolamento de pequeno número de fusobactérias, provàvelmente as amostras não exigentes de proteínas naturais.

  12. Effects of iron-oxide nanoparticles on compound biofilms of streptococcus gordonii and fusobacterium nucleatum

    Science.gov (United States)

    Nguyen, Jane Q.; Withers, Nathan J.; Alas, Gema; Senthil, Arjun; Minetos, Christina; Jaiswal, Nikita; Ivanov, Sergei A.; Huber, Dale L.; Smolyakov, Gennady A.; Osiński, Marek

    2018-02-01

    The human mouth is a host of a large gamut of bacteria species, with over 700 of different bacteria strains identified. Most of these bacterial species are harmless, some are beneficial (such as probiotics assisting in food digestion), but some are responsible for various diseases, primarily tooth decay and gum diseases such as gingivitis and periodontitis. Dental plaque has a complicated structure that varies from patient to patient, but a common factor in most cases is the single species of bacterium acting as a secondary colonizer, namely Fusobacterium nucleatum, while the actual disease is caused by a variety of tertiary colonizers. We hypothesize that destruction of a compound biofilm containing Fusobacterium nucleatum will prevent tertiary colonizers (oral pathogens) from establishing a biofilm, and thus will protect the patient from developing gingivitis and periodontitis. In this paper, we report on the effects of exposure of compound biofilms of a primary colonizer Streptococcus gordonii combined with Fusobacterium nucleatum to iron oxide nanoparticles as possible bactericidal agent.

  13. Mycoplasma pneumoniae preceding Lemierre's syndrome due to Fusobacterium nucleatum complicated by acute Epstein-Barr virus (EBV) infectious mononucleosis in an immunocompetent host.

    Science.gov (United States)

    Klein, Natalie C; Petelin, Andrew; Cunha, Burke A

    2013-01-01

    We report an unusual case of Lemierre's syndrome due to a rare species of Fusobacterium, that is, Fusobacterium nucleatum preceded by Mycoplasma pneumoniae pharyngitis and followed later by Epstein-Barr virus infectious mononucleosis. Copyright © 2013 Elsevier Inc. All rights reserved.

  14. Growth requirements and fermentation products of Fusobacterium prausnitzii, and a proposal to reclassify it as Faecalibacterium prausnitzii gen. nov., comb. nov.

    NARCIS (Netherlands)

    Duncan, SH; Hold, GL; Harmsen, HJM; Stewart, CS; Flint, HJ

    2002-01-01

    Two newly isolated strains of obligately anaerobic bacteria from human faeces are shown here to be related to Fusobacterium prausnitzii, which is regarded as one of the most abundant colonizers of the human colon. These strains, along with Fusobacterium prausnitzii ATCC 27768(T) and 27766, are

  15. Veillonella Catalase Protects the Growth of Fusobacterium nucleatum in Microaerophilic and Streptococcus gordonii-Resident Environments.

    Science.gov (United States)

    Zhou, Peng; Li, Xiaoli; Huang, I-Hsiu; Qi, Fengxia

    2017-10-01

    The oral biofilm is a multispecies community in which antagonism and mutualism coexist among friends and foes to keep an ecological balance of community members. The pioneer colonizers, such as Streptococcus gordonii , produce H 2 O 2 to inhibit the growth of competitors, like the mutans streptococci, as well as strict anaerobic middle and later colonizers of the dental biofilm. Interestingly, Veillonella species, as early colonizers, physically interact (coaggregate) with S. gordonii A putative catalase gene ( catA ) is found in most sequenced Veillonella species; however, the function of this gene is unknown. In this study, we characterized the ecological function of catA from Veillonella parvula PK1910 by integrating it into the only transformable strain, Veillonella atypica OK5, which is catA negative. The strain (OK5- catA ) became more resistant to H 2 O 2 Further studies demonstrated that the catA gene expression is induced by the addition of H 2 O 2 or coculture with S. gordonii Mixed-culture experiments further revealed that the transgenic OK5- catA strain not only enhanced the growth of Fusobacterium nucleatum , a strict anaerobic periodontopathogen, under microaerophilic conditions, but it also rescued F. nucleatum from killing by S. gordonii A potential role of catalase in veillonellae in biofilm ecology and pathogenesis is discussed here. IMPORTANCE Veillonella species, as early colonizers, can coaggregate with many bacteria, including the initial colonizer Streptococcus gordonii and periodontal pathogen Fusobacterium nucleatum , during various stages of oral biofilm formation. In addition to providing binding sites for many microbes, our previous study also showed that Veillonella produces nutrients for the survival and growth of periodontal pathogens. These findings indicate that Veillonella plays an important "bridging" role in the development of oral biofilms and the ecology of the human oral cavity. In this study, we demonstrated that the reducing

  16. Detection of Fusobacterium nucleatum in two cases of empyema and lung abscess using paromomycin-vancomycin supplemented Brucella HK agar.

    Science.gov (United States)

    Nagaoka, Kentaro; Yanagihara, Katsunori; Morinaga, Yoshitomo; Kohno, Shigeru

    2017-02-01

    Fusobacterium nucleatum was found in patients with empyema or pulmonary abscess, using paromomycin-vancomycin Brucella HK agar. In vitro examination revealed that growth of the strains differed significantly in different media. Clinicians should be aware that suboptimal F. nucleatum cultivation methods may result in an underestimation of its frequency. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. [A case of liver abscess caused by Fusobacterium nucleatum in a patient with recurrent periodontal diseases].

    Science.gov (United States)

    Kim, Yong Hwan; Yoon, Hee Jung; Park, Chan Woong; Kim, Jung Ho; Lee, Min Kyung; Kim, Ki Bang; Na, Dong Jib; Kim, Ji Myung

    2011-01-01

    Fusobacteria are anaerobic gram-negative, non-spore forming bacilli found in normal flora of the oral cavity, urogenital tract, and gastrointestinal tract. Fusobacterium nucleatum has been seldom reported as a cause of liver abscess, particularly in immunocompetent hosts. A 55-year-old man with frequent periodontal disease visited our hospital with intermittent fever and headache for 2 months. Abdominal CT scan revealed an 8.2 × 6 cm mass in the right hepatic lobe with central low density. Abscess culture revealed F. nucleatum as the causative organism. Percutaneous abscess drainage and intravenous administration of antibiotics for 4 weeks improved symptoms and decreased the abscess size. We report a rare case of liver abscess due to F. nucleatum in an immunocompetent man with periodontal disease.

  18. Association between Fusobacterium nucleatum and colorectal cancer: Progress and future directions

    Science.gov (United States)

    Zhang, Sheng; Cai, Sanjun; Ma, Yanlei

    2018-01-01

    The initiation and progression of colorectal cancer (CRC) involves genetic and epigenetic alterations influenced by dietary and environmental factors. Increasing evidence has linked the intestinal microbiota and colorectal cancer. More recently, Fusobacterium nucleatum (Fn), an opportunistic commensal anaerobe in the oral cavity, has been associated with CRC. Several research teams have reported an overabundance of Fn in human CRC and have elucidated the possible mechanisms by which Fn is involved in colorectal carcinogenesis in vitro and in mouse models. However, the mechanisms by which Fn promotes colorectal carcinogenesis remain unclear. To provide new perspectives for early diagnosis, the identification of high risk populations and treatment for colorectal cancer, this review will summarize the relative research progresses regarding the relationship between Fn and colorectal cancer. PMID:29760804

  19. Influence of subinhibitory concentrations of antimicrobials on hydrophobicity, adherence and ultra-structure of Fusobacterium nucleatum

    Directory of Open Access Journals (Sweden)

    Okamoto Ana C.

    2002-01-01

    Full Text Available Fusobacterium nucleatum is considered a bridge organism between earlier and later colonizers in dental biofilms and a putative periodontopathogen. In Dentistry, antimicrobial agents are used for treatment and control of infectious diseases associated with dental plaque. Antiseptics have been used in association with antibiotics to reduce infections after oral surgeries. In this study, the influence of subinhibitory concentrations (SC of chlorhexidine, triclosan, penicillin G and metronidazole, on hydrophobicity, adherence to oral epithelial cells, and ultra-structure of F. nucleatum was examined. All isolates were susceptible to chlorhexidine, triclosan, and metronidazole; however, most of the isolates were susceptible to penicillin G, and all of them were hydrophilic when grown with or without antimicrobials. Adherence was decreased by all antimicrobials. Results suggest that adherence of F. nucleatum was influenced by adhesins because structures such as fimbries or capsule were not observed by transmission electronic microscope.

  20. Fournier's gangrene caused by Actinomyces funkei, Fusobacterium gonidiaformans and Clostridium hathewayi.

    Science.gov (United States)

    Tena, Daniel; Losa, Cristina; Medina-Pascual, María José; Sáez-Nieto, Juan Antonio

    2014-06-01

    We report the first case of Fournier's gangrene caused by three unusual anaerobic organisms: Actinomyces funkei, Fusobacterium gonidiaformans and Clostridium hathewayi. The infection occurred in a 73-year-old man without typical risk factors for the development of Fournier's gangrene. Clinical outcome was good after prolonged antibiotic treatment and extensive debridement of the perineum. The case suggests that A. funkei, F. gonidiaformans and C. hathewayi should be considered as potential pathogens of Fournier's gangrene. Human infections caused by these organisms are very rare but can be underestimated because correct identification is very difficult, especially in polymicrobial infections such as Fournier's gangrene. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Crystallization and preliminary X-ray data of the FadA adhesin from Fusobacterium nucleatum

    Energy Technology Data Exchange (ETDEWEB)

    Nithianantham, Stanley [Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4935 (United States); Xu, Minghua [Department of Biological Sciences, School of Dentistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106-4905 (United States); Wu, Nan [Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4935 (United States); Han, Yiping W., E-mail: ywh2@case.edu [Department of Biological Sciences, School of Dentistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106-4905 (United States); Department of Pathology, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States); Shoham, Menachem, E-mail: ywh2@case.edu [Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4935 (United States)

    2006-12-01

    The FadA adhesin from F. nucleatum, which is involved in bacterial attachment and invasion of human oral epithelial cells, has been crystallized in space group P6{sub 1} or P6{sub 5}, and X-ray data have been collected to 1.9 Å resolution. Fusobacterium nucleatum is a Gram-negative anaerobe prevalent in the oral cavity that is associated with periodontal disease, preterm birth and infections in other parts of the human body. The bacteria attach to and invade epithelial and endothelial cells in the gum tissue and elsewhere via a 13.7 kDa adhesin protein FadA (Fusobacterium adhesin A). FadA exists in two forms: the intact form (pre-FadA), consisting of 129 amino acids, and the mature form (mFadA), which lacks an 18-residue signal sequence. Both forms have been expressed in Escherichia coli and purified. mFadA has been crystallized. The crystals belong to the hexagonal space group P6{sub 1} or P6{sub 5}, with unit-cell parameters a = b = 59.3, c = 125.7 Å and one molecule per asymmetric unit. The crystals exhibit an unusually high solvent content of 74%. Synchrotron X-ray data have been collected to 1.9 Å. The crystals are suitable for X-ray structure determination. The crystal structure of FadA may provide a basis for the development of therapeutic agents to combat periodontal disease and other infections associated with F. nucleatum.

  2. Crystallization and preliminary X-ray data of the FadA adhesin from Fusobacterium nucleatum

    International Nuclear Information System (INIS)

    Nithianantham, Stanley; Xu, Minghua; Wu, Nan; Han, Yiping W.; Shoham, Menachem

    2006-01-01

    The FadA adhesin from F. nucleatum, which is involved in bacterial attachment and invasion of human oral epithelial cells, has been crystallized in space group P6 1 or P6 5 , and X-ray data have been collected to 1.9 Å resolution. Fusobacterium nucleatum is a Gram-negative anaerobe prevalent in the oral cavity that is associated with periodontal disease, preterm birth and infections in other parts of the human body. The bacteria attach to and invade epithelial and endothelial cells in the gum tissue and elsewhere via a 13.7 kDa adhesin protein FadA (Fusobacterium adhesin A). FadA exists in two forms: the intact form (pre-FadA), consisting of 129 amino acids, and the mature form (mFadA), which lacks an 18-residue signal sequence. Both forms have been expressed in Escherichia coli and purified. mFadA has been crystallized. The crystals belong to the hexagonal space group P6 1 or P6 5 , with unit-cell parameters a = b = 59.3, c = 125.7 Å and one molecule per asymmetric unit. The crystals exhibit an unusually high solvent content of 74%. Synchrotron X-ray data have been collected to 1.9 Å. The crystals are suitable for X-ray structure determination. The crystal structure of FadA may provide a basis for the development of therapeutic agents to combat periodontal disease and other infections associated with F. nucleatum

  3. Efecto antibacteriano in vitro del colutorio a base de matricaria chamomilla (manzanilla) sobre el fusobacterium nucleatum ATCC 25586

    OpenAIRE

    Delgado Asmat, Edith Esther

    2015-01-01

    This research study was to determine the in vitro antibacterial mouthwash based Matricaria chamomilla (chamomile) Fusobacterium nucleatum against ATCC 25586. The study was conducted in laboratory environments Farmaconogsia of the National University of Trujillo and microbiology laboratories of the Faculty of Medicine of the National University of Trujillo. The sample consisted of 16 discs broadcast, divided into 5 groups; that is, a group with Chlorhexidine samples, chamomile three groups...

  4. Role of Fusobacterium nucleatum and Coaggregation in Anaerobe Survival in Planktonic and Biofilm Oral Microbial Communities during Aeration

    OpenAIRE

    Bradshaw, David J.; Marsh, Philip D.; Watson, G. Keith; Allison, Clive

    1998-01-01

    Coaggregation is a well-characterized phenomenon by which specific pairs of oral bacteria interact physically. The aim of this study was to examine the patterns of coaggregation between obligately anaerobic and oxygen-tolerant species that coexist in a model oral microbial community. Obligate anaerobes other than Fusobacterium nucleatum coaggregated only poorly with oxygen-tolerant species. In contrast, F. nucleatum was able to coaggregate not only with both oxygen-tolerant and other obligate...

  5. Antibacterial effects of Pluchea indica Less leaf extract on E. faecalis and Fusobacterium nucleatum (in vitro

    Directory of Open Access Journals (Sweden)

    Agni Febrina Pargaputri

    2016-06-01

    Full Text Available Background: Enterococcus. faecalis (E. faecalis and Fusobacterium nucleatum (F. nucleatum are the most common bacteria found in infected tooth root canal. Most of these bacteria often cause failure in endodontic treatments. Pluchea indica Less leaf is a species of plants that has several chemical properties. It consists of flavonoids, tannins, polyphenols, and essensial oils which have been reported as antibacterial agents. Because of its benefits, the extract of Pluchea indica Less leaves may be potentially developed as one of root canal sterilization dressing. Purpose: This study aimed to determine antibacterial activity of Pluchea indica Less leaves extract against E. faecalis and F. nucleatum bacteria. Method: Dilution method was conducted first to show Minimum Inhibitory Concentration (MIC of the extract against E. faecalis and F. nucleatum. The antibacterial activity test on Pluchea indica Less leaves extract was performed on E. faecalis and F. nucleatum bacteria using agar diffusion method. The Pluchea indica Less leaves extract used for antibacterial activity test was at a concentrations of 100%, 50%, 25%, 12.5%, and 6.25%. Thirty-five petridiscs were used and divided into five groups based on the extract concentration. Result: The results showed strong and moderate antibacterial effects of the Pluchea indica Less leaves extract on E. faecalis at the concentrations of 100% and 50%, while on F. nucleatum only at the concentration of 100% with moderate effect. Conclusion: Pluchea indica Less leaves extract has antibacterial activity against E. faecalis and F. nucleatum bacteria with strong-moderate effect.

  6. Antimicrobial effect of essential oil of Citrus reticulata on Fusobacterium nucleatum associated with periodontal disease

    Directory of Open Access Journals (Sweden)

    Cindy Giohanna Pardo

    2017-07-01

    Full Text Available Chlorhexidine as a treatment of periodontal disease has achieved bactericidal effects over periodontopathogens and oral biofilm. Its use generates adverse effects; therefore natural alternatives are presented with a similar antimicrobial effect. Essential oils have proved effective in controlling dental plaque without the adverse effects of chlorhexidine. The aim of this study was to determine the bacteriostatic and bactericidal effect of essential oil of tangerine against Fusobacterium nucleatum. The extraction of the essential oil was performed by expression of tangerine peels (Arrayana and Oneco varieties. Concentrations at 20%, 40%, 60%, 80% and 100% of the essential oil diluted in 0,02% Tween were evaluated. The bacteriostatic and bactericidal effect was determined by antimicrobial susceptibility testing by disk diffusion. As a positive control 0,2% chlorhexidine and water as negative control were used. Inhibition zone (mm was measured and presence or absence of bacterial growth was determined from colony forming units.  To compare proportions of bacteriostatic and bactericidal activity, Fisher and T student test (95% CI p = 0,05 were performed. The 100% concentration zone of inhibition showed a similar behavior as chlorhexidine (p 0,05. The use of essential oils of tangerine could be a complementary alternative to treatment of periodontal disease.

  7. Antiaggregation potential of berry fractions against pairs of Streptococcus mutans with Fusobacterium nucleatum or Actinomyces naeslundii.

    Science.gov (United States)

    Riihinen, Kaisu; Ryynänen, Anu; Toivanen, Marko; Könönen, Eija; Törrönen, Riitta; Tikkanen-Kaukanen, Carina

    2011-01-01

    Coaggregation is an interspecies adhesion process, which is essential to the development of dental plaque. This is an in vitro study of the composition of the soluble solids in the berry juice molecular size fractions (100 kDa, FIII) derived from apple, bilberry, blackcurrant, cloudberry, crowberry and lingonberry and their ability to inhibit and reverse coaggregation of the pairs of common species in dental plaque: Streptococcus mutans with Fusobacterium nucleatum or Actinomyces naeslundii. Inhibitory and reversal activity was found in the molecular size fractions FII and FIII of bilberry, blackcurrant, crowberry and lingonberry. The active fractions contained higher amounts of polyphenols (5-12% of soluble solids) than those without activity (juice fractions FII and FIII and also small amounts of anthocyanins were detected. Anthocyanins, proanthocyanidins and flavonol glycosides were prevalent in FII and FIII fractions of bilberry, blackcurrant and crowberry juices. Comparable amounts of sugars and titratable acids were present in the latter three berry juice fractions of different size. The results indicate that the high molecular size fractions of lingonberry, bilberry, blackcurrant and crowberry juices have antiaggregation potential on common oral bacteria, the potential being associated with their polyphenolic content. Copyright © 2010 John Wiley & Sons, Ltd.

  8. Synergistic effect on biofilm formation between Fusobacterium nucleatum and Capnocytophaga ochracea.

    Science.gov (United States)

    Okuda, Tamaki; Okuda, Katsuji; Kokubu, Eitoyo; Kawana, Tomoko; Saito, Atsushi; Ishihara, Kazuyuki

    2012-02-01

    The formation of dental plaque biofilm by specific Gram-negative rods and spirochetes plays an important role in the development of periodontal disease. The aim of this study was to characterize biofilm formation by Fusobacterium nucleatum and Capnocytophaga ochracea. Coaggregation between F. nucleatum and Capnocytophaga species was determined by visual assay. Biofilm formation was assessed by crystal violet staining. Enhancement of biofilm formation by F. nucleatum via soluble factor of C. ochracea was evaluated by addition of culture supernatant and a two-compartment separated co-culture system. Production of autoinducer-2 by the tested organisms was evaluated using Vibrio harveyi BB170. F. nucleatum strains coaggregated with C. ochracea ATCC 33596 or ONO-26 strains. Ethylenediamine tetraacetic acid, N-acetyl-d-galactosamine or lysine inhibited coaggregation. Heating or proteinase K treatment of F. nucleatum cells affected coaggregation, whereas the same treatment of C. ochracea cells did not. Co-culture of F. nucleatum with C. ochracea in the same well resulted in a statistically significant increase in biofilm formation. Enhancement of F. nucleatum biofilm formation by a soluble component of C. ochracea was observed using the two-compartment co-culture system (P culture supernatant of C. ochracea (P < 0.01). The present findings indicate that induction of coaggregation and intracellular interaction by release of a diffusible molecule by C. ochracea play a significant role in the formation of biofilm by F. nucleatum and C. ochracea. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. Effect of kefir on Fusobacterium nucleatum in potentially preventing intestinal cancer

    Directory of Open Access Journals (Sweden)

    Zeynep Banu Guzel-Seydim

    2016-07-01

    Full Text Available Background: Fusobacterium spp. are known to be part of mouth and intestinal microbiota. Fusobacterium nucleatum is an obligate anaerobe, Gram negative, non-spore forming pleomorphic bacillus that can cause diseases not only in the mouth and teeth but also in the brain, pleura, lungs and liver. It was noted that F. nucleatum induces fetal death (fetal demise in pregnant women. Recent studies indicate that F. nucleatum could lead to colon cancer by binding to the epithelial tissue. Kefir is produced from kefir grains that are a source of probiotics. Fermented dairy products and especially kefir and yogurt are significant for functional nutrition. In kefir grains, lactic acid bacteria, acetic acid bacteria and yeasts are embedded in a polysaccharide matrix, called kefiran. When kefir grains are added to milk and incubated for approximately 22 h at 25°C, microorganisms in the grains continue to proliferate in milk with the production of functional metabolic compounds. While yogurt has mainly two bacteria, authentic kefir has its characteristic Lactobacillus kefiranofaciens, Lactobacillus kefir and Lactobacillus kefirgranum, in addition to many other types of lactic acid bacteria (LAB. Previous studies have indicated that fermented dairy products can cause probiotic effects such as improvement in digestive system health, reduction in serum cholesterol, improvement in lactose tolerance, improvement in immune function, control of irritable bowel symptoms, and anticarcinogenic properties. Objective: The aim of this research was to report the effects of fermented dairy products in vitro on the growth of F. nucleatum. Milk, kefir made from natural kefir grains, commercial kefir produced from kefir starter culture, yogurt produced from natural yogurt starter culture and commercial yogurt produced from yogurt starter culture were used against F. nucleatum. Methods: F. nucleatum (ATCC 25586 was grown in Fluid Thioglycollate Medium at 37°C for 3 days under

  10. The proteins of Fusobacterium spp. involved in hydrogen sulfide production from L-cysteine.

    Science.gov (United States)

    Basic, Amina; Blomqvist, Madeleine; Dahlén, Gunnar; Svensäter, Gunnel

    2017-03-14

    Hydrogen sulfide (H 2 S) is a toxic foul-smelling gas produced by subgingival biofilms in patients with periodontal disease and is suggested to be part of the pathogenesis of the disease. We studied the H 2 S-producing protein expression of bacterial strains associated with periodontal disease. Further, we examined the effect of a cysteine-rich growth environment on the synthesis of intracellular enzymes in F. nucleatum polymorphum ATCC 10953. The proteins were subjected to one-dimensional (1DE) and two-dimensional (2DE) gel electrophoresis An in-gel activity assay was used to detect the H 2 S-producing enzymes; Sulfide from H 2 S, produced by the enzymes in the gel, reacted with bismuth forming bismuth sulfide, illustrated as brown bands (1D) or spots (2D) in the gel. The discovered proteins were identified with liquid chromatography - tandem mass spectrometry (LC-MS/MS). Cysteine synthase and proteins involved in the production of the coenzyme pyridoxal 5'phosphate (that catalyzes the production of H 2 S) were frequently found among the discovered enzymes. Interestingly, a higher expression of H 2 S-producing enzymes was detected from bacteria incubated without cysteine prior to the experiment. Numerous enzymes, identified as cysteine synthase, were involved in the production of H 2 S from cysteine and the expression varied among Fusobacterium spp. and strains. No enzymes were detected with the in-gel activity assay among the other periodontitis-associated bacteria tested. The expression of the H 2 S-producing enzymes was dependent on environmental conditions such as cysteine concentration and pH but less dependent on the presence of serum and hemin.

  11. Identification and characterization of fusolisin, the Fusobacterium nucleatum autotransporter serine protease.

    Directory of Open Access Journals (Sweden)

    Lior Doron

    Full Text Available Fusobacterium nucleatum is an oral anaerobe associated with periodontal disease, adverse pregnancy outcomes and colorectal carcinoma. A serine endopeptidase of 61-65 kDa capable of damaging host tissue and of inactivating immune effectors was detected previously in F. nucleatum. Here we describe the identification of this serine protease, named fusolisin, in three oral F. nucleatum sub-species. Gel zymogram revealed fusobacterial proteolytic activity with molecular masses ranging from 55-101 kDa. All of the detected proteases were inhibited by the serine protease inhibitor PMSF. analysis revealed that all of the detected proteases are encoded by genes encoding an open reading frame (ORF with a calculated mass of approximately 115 kDa. Bioinformatics analysis of the identified ORFs demonstrated that they consist of three domains characteristic of autotransporters of the type Va secretion system. Our results suggest that the F. nucleatum fusolisins are derived from a precursor of approximately 115 kDa. After crossing the cytoplasmic membrane and cleavage of the leader sequence, the C-terminal autotransporter domain of the remaining 96-113 kDa protein is embedded in the outer membrane and delivers the N-terminal S8 serine protease passenger domain to the outer cell surface. In most strains the N-terminal catalytic 55-65 kDa domain self cleaves and liberates itself from the autotransporter domain after its transfer across the outer cell membrane. In F. nucleatum ATCC 25586 this autocatalytic activity is less efficient resulting in a full length membrane-anchored serine protease. The mature serine protease was found to cleave after Thr, Gly, Ala and Leu residues at the P1 position. Growth of F. nucleatum in complex medium was inhibited when serine protease inhibitors were used. Additional experiments are needed to determine whether fusolisin might be used as a target for controlling fusobacterial infections.

  12. The danger signal extracellular ATP is an inducer of Fusobacterium nucleatum biofilm dispersal

    Directory of Open Access Journals (Sweden)

    Qinfeng Ding

    2016-11-01

    Full Text Available Plaque biofilm is the primary etiological agent of periodontal disease. Biofilm formation progresses through multiple developmental stages beginning with bacterial attachment to a surface, followed by development of microcolonies and finally detachment and dispersal from a mature biofilm as free planktonic bacteria. Tissue damage arising from inflammatory response to biofilm is one of the hallmark features of periodontal disease. A consequence of tissue damage is the release of ATP from within the cell into the extracellular space. Extracellular ATP (eATP is an example of a danger associated molecular pattern (DAMP employed by mammalian cells to elicit inflammatory and damage healing responses. Although the roles of eATP as a signaling molecule in multi-cellular organisms have been relatively well studied, exogenous ATP also influences bacteria biofilm formation. Since plaque biofilms are continuously exposed to various stresses including exposure to the host damage factors eATP, we hypothesized that eATP, in addition to eliciting inflammation could potentially influence the biofilm lifecycle of periodontal associated bacteria. We found that eATP rather than nutritional factors or oxidative stress induced dispersal of Fusobacterium nucleatum, an organism associated with periodontal disease. eATP induced biofilm dispersal through chelating metal ions present in biofilm. Dispersed F. nucleatum biofilm, regardless of natural or induced dispersal by exogenous ATP, were significantly more adhesive and invasive compared to planktonic or biofilm counterparts, and correspondingly activated significantly more pro-inflammatory cytokine production in infected periodontal fibroblasts. Dispersed F. nucleatum also exhibited significantly higher expression of fadA, a virulence factor implicated in adhesion and invasion, compared to planktonic or biofilm bacteria. This study revealed for the first time that periodontal bacterium is capable of co-opting eATP, a

  13. Streptococcus mutans SpaP binds to RadD of Fusobacterium nucleatum ssp. polymorphum.

    Science.gov (United States)

    Guo, Lihong; Shokeen, Bhumika; He, Xuesong; Shi, Wenyuan; Lux, Renate

    2017-10-01

    Adhesin-mediated bacterial interspecies interactions are important elements in oral biofilm formation. They often occur on a species-specific level, which could determine health or disease association of a biofilm community. Among the key players involved in these processes are the ubiquitous fusobacteria that have been recognized for their ability to interact with numerous different binding partners. Fusobacterial interactions with Streptococcus mutans, an important oral cariogenic pathogen, have previously been described but most studies focused on binding to non-mutans streptococci and specific cognate adhesin pairs remain to be identified. Here, we demonstrated differential binding of oral fusobacteria to S. mutans. Screening of existing mutant derivatives indicated SpaP as the major S. mutans adhesin specific for binding to Fusobacterium nucleatum ssp. polymorphum but none of the other oral fusobacteria tested. We inactivated RadD, a known adhesin of F. nucleatum ssp. nucleatum for interaction with a number of gram-positive species, in F. nucleatum ssp. polymorphum and used a Lactococcus lactis heterologous SpaP expression system to demonstrate SpaP interaction with RadD of F. nucleatum ssp. polymorphum. This is a novel function for SpaP, which has mainly been characterized as an adhesin for binding to host proteins including salivary glycoproteins. In conclusion, we describe an additional role for SpaP as adhesin in interspecies adherence with RadD-SpaP as the interacting adhesin pair for binding between S. mutans and F. nucleatum ssp. polymorphum. Furthermore, S. mutans attachment to oral fusobacteria appears to involve species- and subspecies-dependent adhesin interactions. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Characterization of Fusobacterium varium Fv113-g1 isolated from a patient with ulcerative colitis based on complete genome sequence and transcriptome analysis.

    Directory of Open Access Journals (Sweden)

    Tsuyoshi Sekizuka

    Full Text Available Fusobacterium spp. present in the oral and gut flora is carcinogenic and is associated with the risk of pancreatic and colorectal cancers. Fusobacterium spp. is also implicated in a broad spectrum of human pathologies, including Crohn's disease and ulcerative colitis (UC. Here we report the complete genome sequence of Fusobacterium varium Fv113-g1 (genome size, 3.96 Mb isolated from a patient with UC. Comparative genome analyses totally suggested that Fv113-g1 is basically assigned as F. varium, in particular, it could be reclassified as notable F. varium subsp. similar to F. ulcerans because of partial shared orthologs. Compared with the genome sequences of F. varium ATCC 27725 (genome size, 3.30 Mb and other strains of Fusobacterium spp., Fv113-g1 possesses many accessary pan-genome sequences with noteworthy multiple virulence factors, including 44 autotransporters (type V secretion system, T5SS and 13 Fusobacterium adhesion (FadA paralogs involved in potential mucosal inflammation. Indeed, transcriptome analysis demonstrated that Fv113-g1-specific accessary genes, such as multiple T5SS and fadA paralogs, showed notably increased expression with D-MEM cultivation than with brain heart infusion broth. This implied that growth condition may enhance the expression of such potential virulence factors, leading to remarkable survival against other gut microorganisms and to the pathogenicity to human intestinal epithelium.

  15. Diets That Promote Colon Inflammation Associate With Risk of Colorectal Carcinomas That Contain Fusobacterium nucleatum.

    Science.gov (United States)

    Liu, Li; Tabung, Fred K; Zhang, Xuehong; Nowak, Jonathan A; Qian, Zhi Rong; Hamada, Tsuyoshi; Nevo, Daniel; Bullman, Susan; Mima, Kosuke; Kosumi, Keisuke; da Silva, Annacarolina; Song, Mingyang; Cao, Yin; Twombly, Tyler S; Shi, Yan; Liu, Hongli; Gu, Mancang; Koh, Hideo; Li, Wanwan; Du, Chunxia; Chen, Yang; Li, Chenxi; Li, Wenbin; Mehta, Raaj S; Wu, Kana; Wang, Molin; Kostic, Aleksander D; Giannakis, Marios; Garrett, Wendy S; Hutthenhower, Curtis; Chan, Andrew T; Fuchs, Charles S; Nishihara, Reiko; Ogino, Shuji; Giovannucci, Edward L

    2018-04-24

    Specific nutritional components are likely to induce intestinal inflammation, which is characterized by increased levels of interleukin 6 (IL6), C-reactive protein (CRP), and TNF receptor superfamily member 1B (TNFRSF1B) in the circulation and promotes colorectal carcinogenesis. The inflammatory effects of a diet can be estimated based on empirical dietary inflammatory pattern (EDIP) score, calculated based on intake of 18 foods associated with plasma levels of IL6, CRP, and TNFRSF1B. An inflammatory environment in the colon (based on increased levels of IL6, CRP, and TNFRSF1B in peripheral blood) contributes to impairment of the mucosal barrier and altered immune cell responses, affecting the composition of the intestinal microbiota. Colonization by Fusobacterium nucleatum has been associated with presence and features of colorectal adenocarcinoma. We investigated the association between diets that promote inflammation (based on EDIP score) and colorectal cancer subtypes classified by level of F nucleatum in the tumor microenvironment. We calculated EDIP scores based on answers to questionnaires collected from participants in the Nurses' Health Study (through June 1, 2012) and the Health Professionals Follow-up Study (through January 31, 2012). Participants in both cohorts reported diagnoses of rectal or colon cancer in biennial questionnaires; deaths from unreported colorectal cancer cases were identified through the National Death Index and next of kin. Colorectal tumor tissues were collected from hospitals where the patients underwent tumor resection and F nucleatum DNA was quantified by a PCR assay. We used multivariable duplication-method Cox proportional hazard regression to assess the associations of EDIP scores with risks of colorectal cancer subclassified by F nucleatum status. During 28 years of follow up of 124,433 participants, we documented 951 incident cases of colorectal carcinoma with tissue F nucleatum data. Higher EDIP scores associated with

  16. Transcriptome profiling analysis of senescent gingival fibroblasts in response to Fusobacterium nucleatum infection.

    Directory of Open Access Journals (Sweden)

    Sun-Hee Ahn

    Full Text Available Periodontal disease is caused by dental plaque biofilms. Fusobacterium nucleatum is an important periodontal pathogen involved in the development of bacterial complexity in dental plaque biofilms. Human gingival fibroblasts (GFs act as the first line of defense against oral microorganisms and locally orchestrate immune responses by triggering the production of reactive oxygen species and pro-inflammatory cytokines (IL-6 and IL-8. The frequency and severity of periodontal diseases is known to increase in elderly subjects. However, despite several studies exploring the effects of aging in periodontal disease, the underlying mechanisms through which aging affects the interaction between F. nucleatum and human GFs remain unclear. To identify genes affected by infection, aging, or both, we performed an RNA-Seq analysis using GFs isolated from a single healthy donor that were passaged for a short period of time (P4 'young GFs' or for longer period of time (P22 'old GFs', and infected or not with F. nucleatum. Comparing F. nucleatum-infected and uninfected GF(P4 cells the differentially expressed genes (DEGs were involved in host defense mechanisms (i.e., immune responses and defense responses, whereas comparing F. nucleatum-infected and uninfected GF(P22 cells the DEGs were involved in cell maintenance (i.e., TGF-β signaling, skeletal development. Most DEGs in F. nucleatum-infected GF(P22 cells were downregulated (85% and were significantly associated with host defense responses such as inflammatory responses, when compared to the DEGs in F. nucleatum-infected GF(P4 cells. Five genes (GADD45b, KLF10, CSRNP1, ID1, and TM4SF1 were upregulated in response to F. nucleatum infection; however, this effect was only seen in GF(P22 cells. The genes identified here appear to interact with each other in a network associated with free radical scavenging, cell cycle, and cancer; therefore, they could be potential candidates involved in the aged GF's response to F

  17. Antibody and T cell responses to Fusobacterium nucleatum and Treponema denticola in health and chronic periodontitis.

    Directory of Open Access Journals (Sweden)

    Jieun Shin

    Full Text Available The characteristics of the T cell response to the members of oral flora are poorly understood. We characterized the antibody and T cell responses to FadA and Td92, adhesins from Fusobacterium nucleatum, an oral commensal, and Treponema denticola, a periodontal pathogen, respectively. Peripheral blood and saliva were obtained from healthy individuals and patients with untreated chronic periodontitis (CP, n = 11 paris and after successful treatment of the disease (n = 9. The levels of antigen-specific antibody were measured by ELISA. In plasma, IgG1 was the most abundant isotype of Ab for both Ags, followed by IgA and then IgG4. The levels of FadA-specific salivary IgA (sIgA were higher than Td92-specific sIgA and the FadA-specific IgA levels observed in plasma. However, the periodontal health status of the individuals did not affect the levels of FadA- or Td92-specific antibody. Even healthy individuals contained FadA- and Td92-specific CD4(+ T cells, as determined by the detection of intracytoplasmic CD154 after short-term in vitro stimulation of peripheral blood mononuclear cells (PBMCs with the antigens. Patients with CP tended to possess increased numbers of FadA- and Td92-specific CD4(+ T cells but reduced numbers of Td92-specific Foxp3(+CD4(+ Tregs than the healthy subjects. Both FadA and Td92 induced the production of IFNγ and IL-10 but inhibited the secretion of IL-4 by PBMCs. In conclusion, F. nucleatum induced Th3 (sIgA- and Th1 (IFNγ and IgG1-dominant immune responses, whereas T. denticola induced a Th1 (IFNγ and IgG1-dominant response. This IFNγ-dominant cytokine response was impaired in CP patients, and the Td92-induced IFNγ levels were negatively associated with periodontal destruction in patients. These findings may provide new insights into the homeostatic interaction between the immune system and oral bacteria and the pathogenesis of periodontitis.

  18. Phylogenetic analysis of Fusobacterium prausnitzii based upon the 16S rRNA gene sequence and PCR confirmation.

    Science.gov (United States)

    Wang, R F; Cao, W W; Cerniglia, C E

    1996-01-01

    In order to develop a PCR method to detect Fusobacterium prausnitzii in human feces and to clarify the phylogenetic position of this species, its 16S rRNA gene sequence was determined. The sequence described in this paper is different from the 16S rRNA gene sequence is specific for F. prausnitzii, and the results of this assay confirmed that F. prausnitzii is the most common species in human feces. However, a PCR assay based on the original GenBank sequence was negative when it was performed with two strains of F. prausnitzii obtained from the American Type Culture Collection. A phylogenetic tree based on the new 16S rRNA gene sequence was constructed. On this tree F. prausnitzii was not a member of the Fusobacterium group but was closer to some Eubacterium spp. and located between Clostridium "clusters III and IV" (M.D. Collins, P.A. Lawson, A. Willems, J.J. Cordoba, J. Fernandez-Garayzabal, P. Garcia, J. Cai, H. Hippe, and J.A.E. Farrow, Int. J. Syst. Bacteriol. 44:812-826, 1994).

  19. Rapid brain death caused by a cerebellar abscess with Fusobacterium nucleatum in a young man with drug abuse: a case report.

    Science.gov (United States)

    Hischebeth, Gunnar T R; Keil, Vera C; Gentil, Katrin; Boström, Azize; Kuchelmeister, Klaus; Bekeredjian-Ding, Isabelle

    2014-06-10

    Fusobacterium nucleatum is a strict anaerobic microorganism that causes disease entities such as periodontal and soft tissue abscesses, pulmonary and intraabdominal infections and very rarely intracerebral infections. Here, we report the rare case of a previously healthy 25-year-old German man with a cerebellar abscess caused by Fusobacterium nucleatum that resulted in rapid brain death. Toxicological screening showed positivity for amphetamines and cannabis. The diagnosis was obtained by polymerase chain reaction amplification of bacterial deoxyribonucleic acid in cerebrospinal fluid. In drug users clinicians should think about rare causes of brain abscesses/meningitis. Early diagnosis is necessary and justifies the use of molecular techniques.

  20. Bad news itself or just the messenger? The high mortality of Fusobacterium spp. infections is related to disseminated malignancy and other comorbidities

    DEFF Research Database (Denmark)

    Johannesen, Katrine; Dessau, Ram Benny; Heltberg, Ole

    2016-01-01

    Background Fusobacterium species are pleomorphic, obligate anaerobic gram-negative bacilli. They are difficult to culture and grow slowly. If antibiotic treatment is initiated prior to blood cultures, the bacteria might evade detection. This is a comprehensive report on mortality in non-bacteraem...

  1. Genetic diversity of oral Fusobacterium nucleatum isolated from patients with different clinical conditions Diversidade genética de Fusobacterium nucleatum orais isolados de pacientes com diferentes condições clínicas

    Directory of Open Access Journals (Sweden)

    Mario J. Avila-Campos

    2006-04-01

    Full Text Available The genetic diversity of 23 oral Fusobacterium nucleatum isolated from 15 periodontal patients, eight from seven healthy subjects, nine from nine AIDS patients and two from two Cebus apella monkeys were analyzed. EcoRI restricted the bacterial DNA and 28 ribotypes grouped from A to J groups were obtained. Isolates formed 24 ribotypes which were contained into A, B, C, D, E and F groups, and three reference strains and two clinical isolates of A. actinomycetemcomitans, and E. coli CDC formed four different ribotypes into the G, H, I and J groups. Moreover, from nine F. nucleatum from AIDS patients, six were ribotyped as group C and three as group D. By using ribotyping we distinguished F. nucleatum recovered from different sources. It is possible that isolates from AIDS patients may contain some phenotypic or genotypic factor did not observed in this study.Neste estudo foi avaliada a diversidade genética de 23 amostras de Fusobacterium nucleatum isoladas da cavidade bucal de 15 pacientes com doença periodontal, de oito cepas isoladas de sete indivíduos sadios, de nove isoladas de nove pacientes com AIDS e de duas isoladas de dois macacos Cebus apella. Pela ação da enzima EcoRI sobre o DNA bacteriano foram reconhecidos 28 ribotipos agrupados de A a J. Os isolados testados formaram 24 ribotipos os quais foram contidos nos grupos A, B, C, D, E e F, e as três cepas de referência e dois isolados clínicos de A. actinomycetemcomitans e E. coli CDC formaram quatro diferentes ribotipos contidos nos grupos G, H, I e J. Em adição, as nove cepas de F. nucleatum isoladas de pacientes com AIDS, seis pertenciam ao grupo C e três ao grupo D. Usando-se a ribotipagem foi possível distinguir F. nucleatum isolados de diferentes origens.

  2. Fusobacterium nucleatum-Induced Impairment of Autophagic Flux Enhances the Expression of Proinflammatory Cytokines via ROS in Caco-2 Cells.

    Directory of Open Access Journals (Sweden)

    Bin Tang

    Full Text Available Fusobacterium nucleatum (F. nucleatum plays a critical role in gastrointestinal inflammation. However, the exact mechanism by which F. nucleatum contributes to inflammation is unclear. In the present study, it was revealed that F. nucleatum could induce the production of proinflammatory cytokines (IL-8, IL-1β and TNF-α and reactive oxygen species (ROS in Caco-2 colorectal adenocarcinoma cells. Furthermore, ROS scavengers (NAC or Tiron could decrease the production of proinflammatory cytokines during F. nucleatum infection. In addition, we observed that autophagy is impaired in Caco-2 cells after F. nucleatum infection. The production of proinflammatory cytokines and ROS induced by F. nucleatum was enhanced with either autophagy pharmacologic inhibitors (3-methyladenine, bafilomycin A1 or RNA interference in essential autophagy genes (ATG5 or ATG12 in Caco-2 cells. Taken together, these results indicate that F. nucleatum-induced impairment of autophagic flux enhances the expression of proinflammatory cytokines via ROS in Caco-2 Cells.

  3. Presence and localization of bacteria in the bovine endometrium postpartum using fluorescence in situ hybridization

    DEFF Research Database (Denmark)

    Karstrup, C. C.; Agerholm, J. S.; Jensen, Tim Kåre

    2017-01-01

    The aim of this study was to investigate bacterial invasiveness of the bovine endometrium during the postpartum period. Fluorescence in situ hybridization was applied to endometrial biopsies using probes for Fusobacterium necrophorum, Porphyromonas levii, Trueperella pyogenes, Escherichia coli...

  4. 21 CFR 522.2260 - Sulfamethazine.

    Science.gov (United States)

    2010-04-01

    ..., thereafter. (2) Indications for use. For cattle for treatment of bacterial pneumonia and bovine respiratory... (Fusobacterium necrophorum), acute mastitis and acute metritis (Streptococcus spp.) when caused by one or more...

  5. Bacteriocin-like activity of oral Fusobacterium nucleatum isolated from human and non-human primates Atividade semelhante a bacteriocina de Fusobacterium nucleatum orais isolados de primatas humanos e não-humanos

    Directory of Open Access Journals (Sweden)

    Elerson Gaetti-Jardim Júnior

    1999-12-01

    Full Text Available Fusobacterium nucleatum is indigenous of the human oral cavity and has been involved in different infectious processes. The production of bacteriocin-like substances may be important in regulation of bacterial microbiota in oral cavity. The ability to produce bacteriocin-like substances by 80 oral F. nucleatum isolates obtained from periodontal patients, healthy individuals and Cebus apella monkeys, was examinated. 17.5% of all tested isolates showed auto-antagonism and 78.8% iso- or hetero-antagonism. No isolate from monkey was capable to produce auto-inhibition. In this study, the antagonistic substances production was variable in all tested isolates. Most of the F. nucleatum showed antagonistic activity against tested reference strains. These data suggest a possible participation of these substances on the oral microbial ecology in humans and animals. However, the role of bacteriocins in regulating dental plaque microbiota in vivo is discussed.Fusobacterium nucleatum é indígena da cavidade oral humana e tem sido envolvido em diferentes processos infecciosos. A produção de substâncias semelhantes a bacteriocinas pode ser importante na regulação da microbiota bacteriana da cavidade oral. A capacidade de produzir substâncias tipo bacteriocina de 80 isolados de F. nucleatum orais, obtidos de pacientes com doença periodontal, indivíduos sadios e macaco Cebus apella, foi avaliada. 17,5% de todos os isolados mostrou auto-antagonismo e 78,8% iso- ou hetero-antagonismo. Nenhum isolado de macaco foi capaz de produzir auto-inibição. Neste estudo, a produção de substâncias antagonístas foi variável em todos os isolados testados. A maioria dos F. nucleatum mostrou atividade antagonísta para as cepas de referência testadas. Esses dados sugerem a possível participação dessas substâncias sobre a ecologia microbiana em humanos e animais. Entretanto, o papel das bacteriocinas na regulação da microbiota da placa dental in vivo

  6. TLR2/TLR4 activation induces Tregs and suppresses intestinal inflammation caused by Fusobacterium nucleatum in vivo.

    Directory of Open Access Journals (Sweden)

    Yin-Ping Jia

    Full Text Available Toll-like receptors (TLRs 2 and 4 play critical roles in intestinal inflammation caused by Fusobacterium nucleatum (F. nucleatum infection, but the role of TLR2/TLR4 in regulation of proinflammatory cytokines remains unknown. In this study, through microarray analysis and qRT-PCR, we showed that TLR2/TLR4 are involved in the F. nucleatum-induced inflammatory signaling pathway in Caco-2 cells, C57BL/6 mice and human clinical specimens. In TLR2-/- and TLR4-/- mice, F. nucleatum infection resulted in increased colonization of the bacteria and production of the proinflammatory cytokines IL-8, IL-1β and TNF-α. In addition, the ratio of Foxp3+ CD4+ T cells in the total CD4+ T cells in TLR2-/- and TLR4-/- mice was less than that in wild-type mice, and the ratio in hybrid mice was more than that in knockout mice, which suggested that TLR2/TLR4 mediated the number of Tregs. Furthermore, it was observed that inflammatory cytokine levels were reduced in TLR2-/- mice after Treg transfer. Thus, these data indicate that TLR2/TLR4 regulate F. nucleatum-induced inflammatory cytokines through Tregs in vivo.

  7. Role of Fusobacterium nucleatum and Coaggregation in Anaerobe Survival in Planktonic and Biofilm Oral Microbial Communities during Aeration

    Science.gov (United States)

    Bradshaw, David J.; Marsh, Philip D.; Watson, G. Keith; Allison, Clive

    1998-01-01

    Coaggregation is a well-characterized phenomenon by which specific pairs of oral bacteria interact physically. The aim of this study was to examine the patterns of coaggregation between obligately anaerobic and oxygen-tolerant species that coexist in a model oral microbial community. Obligate anaerobes other than Fusobacterium nucleatum coaggregated only poorly with oxygen-tolerant species. In contrast, F. nucleatum was able to coaggregate not only with both oxygen-tolerant and other obligately anaerobic species but also with otherwise-noncoaggregating obligate anaerobe–oxygen-tolerant species pairs. The effects of the presence or absence of F. nucleatum on anaerobe survival in both the biofilm and planktonic phases of a complex community of oral bacteria grown in an aerated (gas phase, 200 ml of 5% CO2 in air · min−1) chemostat system were then investigated. In the presence of F. nucleatum, anaerobes persisted in high numbers (>107 · ml−1 in the planktonic phase and >107 · cm−2 in 4-day biofilms). In an equivalent culture in the absence of F. nucleatum, the numbers of black-pigmented anaerobes (Porphyromonas gingivalis and Prevotella nigrescens) were significantly reduced (P ≤ 0.001) in both the planktonic phase and in 4-day biofilms, while the numbers of facultatively anaerobic bacteria increased in these communities. Coaggregation-mediated interactions between F. nucleatum and other species facilitated the survival of obligate anaerobes in aerated environments. PMID:9746571

  8. Biochemical characterization of a novel tyrosine phenol-lyase from Fusobacterium nucleatum for highly efficient biosynthesis of l-DOPA.

    Science.gov (United States)

    Zheng, Ren-Chao; Tang, Xiao-Ling; Suo, Hui; Feng, Li-Lin; Liu, Xiao; Yang, Jian; Zheng, Yu-Guo

    2018-05-01

    Tyrosine phenol-lyase (TPL) catalyzes the reversible cleavage of l-tyrosine to phenol, pyruvate and ammonia. When pyrocatechol is substituted for phenol, l-dihydroxyphenylalanine (l-DOPA) is produced. The TPL-catalyzed route was regarded as the most economic process for l-DOPA production. In this study, a novel TPL from Fusobacterium nucleatum (Fn-TPL) was successfully overexpressed in Escherichia coli and screened for l-DOPA synthesis with a specific activity of 2.69Umg -1 . Fn-TPL was found to be a tetramer, and the optimal temperature and pH for α, β-elimination of l-tyrosine was 60°C and pH 8.5, respectively. The enzyme showed broad substrate specificity toward natural and synthetic l-amino acids. Kinetic analysis suggested that the k cat /K m value for l-tyrosine decomposition was much higher than that for l-DOPA decomposition, while Fn-TPL exhibited similar catalytic efficiency for synthesis of l-tyrosine and l-DOPA. With whole cells of recombinant E. coli as biocatalyst, l-DOPA yield reached 110gL -1 with a pyrocatechol conversion of 95%, which was comparable to the reported highest level. The results demonstrated the great potential of Fn-TPL for industrial production of l-DOPA. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Quantitative proteomic analysis of extracellular matrix extracted from mono- and dual-species biofilms of Fusobacterium nucleatum and Porphyromonas gingivalis.

    Science.gov (United States)

    Mohammed, Marwan Mansoor Ali; Pettersen, Veronika Kuchařová; Nerland, Audun H; Wiker, Harald G; Bakken, Vidar

    2017-04-01

    The Gram-negative bacteria Fusobacterium nucleatum and Porphyromonas gingivalis are members of a complex dental biofilm associated with periodontal disease. In this study, we cultured F. nucleatum and P. gingivalis as mono- and dual-species biofilms, and analyzed the protein composition of the biofilms extracellular polymeric matrix (EPM) by high-resolution liquid chromatography-tandem mass spectrometry. Label-free quantitative proteomic analysis was used for identification of proteins and sequence-based functional characterization for their classification and prediction of possible roles in EPM. We identified 542, 93 and 280 proteins in the matrix of F. nucleatum, P. gingivalis, and the dual-species biofilm, respectively. Nearly 70% of all EPM proteins in the dual-species biofilm originated from F. nucleatum, and a majority of these were cytoplasmic proteins, suggesting an enhanced lysis of F. nucleatum cells. The proteomic analysis also indicated an interaction between the two species: 22 F. nucleatum proteins showed differential levels between the mono and dual-species EPMs, and 11 proteins (8 and 3 from F. nucleatum and P. gingivalis, respectively) were exclusively detected in the dual-species EPM. Oxidoreductases and chaperones were among the most abundant proteins identified in all three EPMs. The biofilm matrices in addition contained several known and hypothetical virulence proteins, which can mediate adhesion to the host cells and disintegration of the periodontal tissues. This study demonstrated that the biofilm matrix of two important periodontal pathogens consists of a multitude of proteins whose amounts and functionalities vary largely. Relatively high levels of several of the detected proteins might facilitate their potential use as targets for the inhibition of biofilm development. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Association of Dietary Patterns With Risk of Colorectal Cancer Subtypes Classified by Fusobacterium nucleatum in Tumor Tissue.

    Science.gov (United States)

    Mehta, Raaj S; Nishihara, Reiko; Cao, Yin; Song, Mingyang; Mima, Kosuke; Qian, Zhi Rong; Nowak, Jonathan A; Kosumi, Keisuke; Hamada, Tsuyoshi; Masugi, Yohei; Bullman, Susan; Drew, David A; Kostic, Aleksandar D; Fung, Teresa T; Garrett, Wendy S; Huttenhower, Curtis; Wu, Kana; Meyerhardt, Jeffrey A; Zhang, Xuehong; Willett, Walter C; Giovannucci, Edward L; Fuchs, Charles S; Chan, Andrew T; Ogino, Shuji

    2017-07-01

    Fusobacterium nucleatum appears to play a role in colorectal carcinogenesis through suppression of the hosts' immune response to tumor. Evidence also suggests that diet influences intestinal F nucleatum. However, the role of F nucleatum in mediating the relationship between diet and the risk of colorectal cancer is unknown. To test the hypothesis that the associations of prudent diets (rich in whole grains and dietary fiber) and Western diets (rich in red and processed meat, refined grains, and desserts) with colorectal cancer risk may differ according to the presence of F nucleatum in tumor tissue. A prospective cohort study was conducted using data from the Nurses' Health Study (June 1, 1980, to June 1, 2012) and the Health Professionals Follow-up Study (June 1, 1986, to June 1, 2012) on a total of 121 700 US female nurses and 51 529 US male health professionals aged 30 to 55 years and 40 to 75 years, respectively (both predominantly white individuals), at enrollment. Data analysis was performed from March 15, 2015, to August 10, 2016. Prudent and Western diets. Incidence of colorectal carcinoma subclassified by F nucleatum status in tumor tissue, determined by quantitative polymerase chain reaction. Of the 173 229 individuals considered for the study, 137 217 were included in the analysis, 47 449 were male (34.6%), and mean (SD) baseline age for men was 54.0 (9.8) years and for women, 46.3 (7.2) years. A total of 1019 incident colon and rectal cancer cases with available F nucleatum data were documented over 26 to 32 years of follow-up, encompassing 3 643 562 person-years. The association of prudent diet with colorectal cancer significantly differed by tissue F nucleatum status (P = .01 for heterogeneity); prudent diet score was associated with a lower risk of F nucleatum-positive cancers (P = .003 for trend; multivariable hazard ratio of 0.43; 95% CI, 0.25-0.72, for the highest vs the lowest prudent score quartile) but not with F nucleatum

  11. Detection of fusobacterium nucleatum and fadA adhesin gene in patients with orthodontic gingivitis and non-orthodontic periodontal inflammation.

    Science.gov (United States)

    Liu, Ping; Liu, Yi; Wang, Jianning; Guo, Yang; Zhang, Yujie; Xiao, Shuiqing

    2014-01-01

    Fusobacterium nucleatum is one of the most abundant gram-negative bacilli colonizing the subgingival plaque and closely associated with periodontal disease. However it is unclear whether F. nucleatum is involved in gingival inflammation under orthodontic appliance. A novel adhesin, FadA, which is unique to oral Fusobacteria, is required for F. nucleatum binding and invasion to epithelial cells and thus may play an important role in colonization of Fusobacterium in the host. In this study, we evaluated the prevalence of F. nucleatum and its virulence factor FadA adhesion gene (fadA) in 169 subgingival biofilm samples from 55 cases of gingivitis patients with orthodontic appliances, 49 cases of gingivitis patients without orthodontic treatment, 35 cases of periodontitis patients and 30 cases of periodontally healthy people via PCR. The correlations between the F. nucleatum/fadA and gingivitis index(GI)was also analyzed. The detection rate of F. nucleatum/fadA in periodontitis group and non-orthodontic gingivitis group was higher than the other two groups (pgingivitis group than in health people (pgingivitis and periodontal disease compared with orthodontic gingivitis.

  12. Effects of methyl gallate and gallic acid on the production of inflammatory mediators interleukin-6 and interleukin-8 by oral epithelial cells stimulated with Fusobacterium nucleatum.

    Science.gov (United States)

    Kang, Mi-Sun; Jang, Hee-Sook; Oh, Jong-Suk; Yang, Kyu-Ho; Choi, Nam-Ki; Lim, Hoi-Soon; Kim, Seon-Mi

    2009-12-01

    Interactions between periodontal bacteria and human oral epithelial cells can lead to the activation and expression of a variety of inflammatory mediators in epithelial cells. Fusobacterium nucleatum is a filamentous human pathogen that is strongly associated with periodontal diseases. This study examined the effects of methyl gallate (MG) and gallic acid (GA) on the production of inflammatory mediators, interleukin (IL)-6 and IL-8, by oral epithelial cells stimulated by F. nucleatum. In a real-time reverse transcription-polymerase chain reaction and an enzyme-linked immunosorbent assay, live F. nucleatum induced high levels of gene expression and protein release of IL-6 and IL-8. The effects of MG and GA were examined by treating KB oral epithelial cells with MG and GA and stimulating them with F. nucleatum. MG and GA inhibited significantly the increases in the IL-6 and IL-8 gene and protein levels in a dose-dependent manner. These Compounds also inhibited the growth of F. nucleatum. No visible effects of MG and GA on the adhesion and invasion of KB cells by F. nucleatum were observed. In conclusion, both MG and GA inhibit IL-6 and IL-8 production from F. nucleatum-activated KB cells.

  13. Progression of periodontal inflammation in adolescents is associated with increased number of Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Fusobacterium nucleatum.

    Science.gov (United States)

    Yang, Ning-Yan; Zhang, Quan; Li, Jin-Lu; Yang, Sheng-Hui; Shi, Qing

    2014-05-01

    The study aims to evaluate the change of related subgingival periodontopathogens among different stage of gingivitis in adolescent and assess the relationship between periodontopathogens and the progression of periodontal inflammation. A total of 77 subgingival plaque samples from 35 adolescent individuals were divided into three groups including gingivitis group (mild, 15 samples; moderate, 16 samples; severe, 15 samples), chronic periodontitis group (15 samples) and healthy group (15 samples). Real-time PCR was used to quantitate Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Fusobacterium nucleatum in subgingival plaque samples. All species, except for F. nucleatum, were detected in samples from gingivitis and periodontitis groups in significantly greater number than in those from healthy group (P periodontitis group in significantly greater number than in those from moderate-to-severe gingivitis group (P periodontal inflammation in adolescents. Examination of periodontopathogens number in adolescents may be of some value for monitoring of periodontal disease development. © 2013 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Fusobacterium nucleatum Alters Atherosclerosis Risk Factors and Enhances Inflammatory Markers with an Atheroprotective Immune Response in ApoE(null Mice.

    Directory of Open Access Journals (Sweden)

    Irina M Velsko

    Full Text Available The American Heart Association supports an association between periodontal disease (PD and atherosclerotic vascular disease (ASVD but does not as of yet support a causal relationship. Recently, we have shown that major periodontal pathogens Porphyromonas gingivalis and Treponema denticola are causally associated with acceleration of aortic atherosclerosis in ApoEnull hyperlipidemic mice. The aim of this study was to determine if oral infection with another significant periodontal pathogen Fusobacterium nucleatum can accelerate aortic inflammation and atherosclerosis in the aortic artery of ApoEnull mice. ApoEnull mice (n = 23 were orally infected with F. nucleatum ATCC 49256 and euthanized at 12 and 24 weeks. Periodontal disease assessments including F. nucleatum oral colonization, gingival inflammation, immune response, intrabony defects, and alveolar bone resorption were evaluated. Systemic organs were evaluated for infection, aortic sections were examined for atherosclerosis, and inflammatory markers were measured. Chronic oral infection established F. nucleatum colonization in the oral cavity, induced significant humoral IgG (P=0.0001 and IgM (P=0.001 antibody response (12 and 24 weeks, and resulted in significant (P=0.0001 alveolar bone resorption and intrabony defects. F. nucleatum genomic DNA was detected in systemic organs (heart, aorta, liver, kidney, lung indicating bacteremia. Aortic atherosclerotic plaque area was measured and showed a local inflammatory infiltrate revealed the presence of F4/80+ macrophages and CD3+ T cells. Vascular inflammation was detected by enhanced systemic cytokines (CD30L, IL-4, IL-12, oxidized LDL and serum amyloid A, as well as altered serum lipid profile (cholesterol, triglycerides, chylomicrons, VLDL, LDL, HDL, in infected mice and altered aortic gene expression in infected mice. Despite evidence for systemic infection in several organs and modulation of known atherosclerosis risk factors, aortic

  15. Fusobacterium nucleatum binding to complement regulatory protein CD46 modulates the expression and secretion of cytokines and matrix metalloproteinases by oral epithelial cells.

    Science.gov (United States)

    Mahtout, Hayette; Chandad, Fatiha; Rojo, Jose M; Grenier, Daniel

    2011-02-01

    Periodontitis is a chronic inflammatory disease that results in the destruction of the supporting tissues of the teeth. Gingival epithelial cells are an important mechanical barrier and participate in the host inflammatory response to periodontopathogens. The aim of the present study is to investigate the capacity of Fusobacterium nucleatum to bind to the complement regulatory protein CD46 expressed by oral epithelial cells and to determine the impact of the binding on the gene expression and protein secretion of interleukin (IL)-6, IL-8, and matrix metalloproteinase (MMP)-9 by oral epithelial cells. Binding of recombinant human CD46 to the surface of F. nucleatum was demonstrated by immunologic assays. After stimulation of oral epithelial cells with F. nucleatum, gene expression was determined by real-time polymerase chain reaction analysis while protein secretion was monitored by enzyme-linked immunosorbent assays. Heat and protease treatments of bacterial cells reduced CD46 binding. F. nucleatum-bound CD46 mediated the cleavage of C3b in the presence of factor I. Stimulating oral epithelial cells with F. nucleatum at a multiplicity of infection of 50 resulted in a significant upregulation of the gene expression and protein secretion of IL-6, IL-8, and MMP-9 by oral epithelial cells. However, pretreating the epithelial cells with an anti-CD46 polyclonal antibody attenuated the production of IL-6, IL-8, and MMP-9 in response to F. nucleatum. Such an inhibitory effect was not observed with non-specific antibodies. The present study demonstrates that F. nucleatum can bind the complement regulatory protein CD46. The interaction of F. nucleatum with epithelial cell surface CD46 may contribute to increasing the levels of proinflammatory mediators and MMPs in periodontal sites and consequently modulate tissue destruction.

  16. Wild Blueberry (Vaccinium angustifolium Ait.) Polyphenols Target Fusobacterium nucleatum and the Host Inflammatory Response: Potential Innovative Molecules for Treating Periodontal Diseases.

    Science.gov (United States)

    Ben Lagha, Amel; Dudonné, Stéphanie; Desjardins, Yves; Grenier, Daniel

    2015-08-12

    Blueberries contain significant amounts of flavonoids to which a number of beneficial health effects in humans have been associated. The present study investigated the effect of a polyphenol-rich lowbush blueberry (Vaccinium angustifolium Ait.) extract on the two main etiologic components of periodontitis, a multifactorial disorder affecting the supporting structures of the teeth. Phenolic acids, flavonoids (flavonols, anthocyanins, flavan-3-ols), and procyanidins made up 16.6, 12.9, and 2.7% of the blueberry extract, respectively. The blueberry extract showed antibacterial activity (MIC = 1 mg/mL) against the periodontopathogenic bacterium Fusobacterium nucleatum. This property may result from the ability of blueberry polyphenols to chelate iron. Moreover, the blueberry extract at 62.5 μg/mL inhibited F. nucleatum biofilm formation by 87.5 ± 2.3%. Subsequently, the ability of the blueberry extract to inhibit the NF-κB signaling pathway in U937-3xκB cells was investigated. The blueberry extract dose-dependently inhibited the activation of NF-κB induced by F. nucleatum. In addition, a pretreatment of macrophages with the blueberry extract (62.5 μg/mL) inhibited the secretion of IL-1β, TNF-α, and IL-6 by 87.3 ± 1.3, 80.7 ± 5.6, and 28.2 ± 9.3%, respectively, following a stimulation with F. nucleatum. Similarly, the secretion of MMP-8 and MMP-9 was also dose-dependently inhibited. This dual antibacterial and anti-inflammatory action of lowbush blueberry polyphenols suggests that they may be promising candidates for novel therapeutic agents.

  17. Stimulation of epithelial cell matrix metalloproteinase (MMP-2, -9, -13) and interleukin-8 secretion by fusobacteria.

    Science.gov (United States)

    Gursoy, U K; Könönen, E; Uitto, V-J

    2008-10-01

    Bacterial pathogens involved in periodontal diseases exert their destructive effects primarily by stimulating the host cells to increase their secretion of proinflammatory cytokines and matrix metalloproteinases (MMPs). This study aimed to determine the epithelial cell matrix metalloproteinase and interleukin-8 (IL-8) secretion upon exposure to fusobacteria. Eight different oral and non-oral Fusobacterium strains were incubated with HaCaT epithelial cells. Gelatin zymography and Western blot analysis were performed to detect collagenase 3 (MMP-13), gelatinase A (MMP-2), gelatinase B (MMP-9), and IL-8 secretion by epithelial cells. All Fusobacterium strains, especially Fusobacterium necrophorum ATCC 25286, Fusobacterium nucleatum ATCC 25586, and Fusobacterium varium ATCC 51644, increased MMP-9 and MMP-13 secretion. Fusobacterium simiae ATCC 33568, and to a lesser extent F. nucleatum and F. necrophorum, increased epithelial MMP-2 secretion. F. nucleatum and F. necrophorum also increased IL-8 secretion. F. varium ATCC 27725, a strain that only weakly stimulated MMP production, strongly increased the IL-8 production, suggesting that their expression is differently regulated. We conclude that the pathogenic potential of fusobacteria may partly result from their ability to stimulate secretion of MMP-9, MMP-13, and IL-8 from epithelial cells.

  18. Cell Based Drug Delivery: Micrococcus luteus Loaded Neutrophils as Chlorhexidine Delivery Vehicles in a Mouse Model of Liver Abscesses in Cattle.

    Science.gov (United States)

    Wendel, Sebastian O; Menon, Sailesh; Alshetaiwi, Hamad; Shrestha, Tej B; Chlebanowski, Lauren; Hsu, Wei-Wen; Bossmann, Stefan H; Narayanan, Sanjeev; Troyer, Deryl L

    2015-01-01

    The recent WHO report on antibiotic resistances shows a dramatic increase of microbial resistance against antibiotics. With only a few new antibiotics in the pipeline, a different drug delivery approach is urgently needed. We have obtained evidence demonstrating the effectiveness of a cell based drug delivery system that utilizes the innate immune system as targeting carrier for antibacterial drugs. In this study we show the efficient loading of neutrophil granulocytes with chlorhexidine and the complete killing of E. coli as well as Fusobacterium necrophorum in in-vitro studies. Fusobacterium necrophorum causes hepatic abscesses in cattle fed high grain diets. We also show in a mouse model that this delivery system targets infections of F. necrophorum in the liver and reduces the bacterial burden by an order of magnitude from approximately 2•106 to 1•105.

  19. Laminitis and interdigital dermatitis and heel horn erosion. A European perspective.

    Science.gov (United States)

    Peterse, D J

    1985-03-01

    Laminitis is one of the most important claw disorders in dairy herds. Nutrition, calving, burdening of the lateral claw of the rear feet, and hereditary susceptibility are all contributing factors. Interdigital dermatitis in cattle may be a result of infection by Bacteroides nodosus and Fusobacterium necrophorum. If this infection becomes chronic, heel horn erosion is its consequence.

  20. Throat ache ans swelling of the neck: first symptoms of Lemierre's syndrome

    NARCIS (Netherlands)

    de Lange, J.; Ybema, A; Baas, E. M.

    2014-01-01

    Lemierre's syndrome, a thrombophlebitis of the internal jugular vein, is a rare disorder, usually caused by the microorganism Fusobacterium necrophorum. Throat ache and swelling of the neck are often the first symptoms. Without adequate treatment, Lemierre's syndrome may result in thrombosis of the

  1. Direct 16S rRNA gene sequencing of polymicrobial culture-negative samples with analysis of mixed chromatograms

    DEFF Research Database (Denmark)

    Hartmeyer, Gitte N; Justesen, Ulrik S

    2010-01-01

    Two cases involving polymicrobial culture-negative samples were investigated by 16S rRNA gene sequencing, with analysis of mixed chromatograms. Fusobacterium necrophorum, Prevotella intermedia and Streptococcus constellatus were identified from pleural fluid in a patient with Lemierre's syndrome...

  2. Behandling af peritonsillær absces

    DEFF Research Database (Denmark)

    Anthonsen, Kristian; Trolle, Waldemar

    2012-01-01

    Peritonsillar abscess (PTA) is associated with high morbidity and can cause serious and life-threatening complications. In Denmark, the most commonly isolated bacteria are Fusobacterium necrophorum and Streptococcus pyogenes gr. A. The incidence of PTA in Denmark is 41/100,000/year, the highest...

  3. Insights into the microbiota of the bovine uterus

    DEFF Research Database (Denmark)

    Rødtness Vesterby Knudsen, Lif; Christensen Karstrup, Cecilia; Gervi Pedersen, Hanne

    (Machado et al. 2012 and Galvão et al. 2012) while Escherichia coli, Trueperella Pyogenes, Prevotella species and Fusobacterium necrophorum have commonly been associated with endometritis (Sheldon 2006). Preliminary results indicate that we often find bacteria from phylum Actinobacteria in the healthy cows...

  4. Lemierre syndrome presenting as acute mastoiditis in a 2-year-old girl with congenital dwarfism

    Directory of Open Access Journals (Sweden)

    Jason B. Fischer

    2015-06-01

    Full Text Available Lemierre syndrome is defined by septic thrombophlebitis of the internal jugular vein caused by Fusobacterium. Historically, these infections originate from the oropharynx and typically are seen in older children, adolescents and young adults. More recently, otogenic sources in younger children have been described with increasing frequency. We present a case of a two-year old, who initially developed an otitis media with perforation of the tympanic membrane and went on to develop mastoiditis and non-occlusive thrombosis of the venous sinus and right internal jugular vein. Fusobacterium necrophorum was grown from operative cultures of the mastoid, ensuing computed tomography scan revealed occlusion of the internal jugular vein and the patient was successfully treated with clindamycin, ciprofloxacin and enoxaparin. This case demonstrates the importance of considering Fusobacterium in otogenic infections and the consideration of Lemierre syndrome when F. necrophorum is identified.

  5. High-resolution microbiome profiling uncovers Fusobacterium nucleatum, Lactobacillus gasseri/johnsonii, and Lactobacillus vaginalis associated to oral and oropharyngeal cancer in saliva from HPV positive and HPV negative patients treated with surgery and chemo-radiation.

    Science.gov (United States)

    Guerrero-Preston, Rafael; White, James Robert; Godoy-Vitorino, Filipa; Rodríguez-Hilario, Arnold; Navarro, Kelvin; González, Herminio; Michailidi, Christina; Jedlicka, Anne; Canapp, Sierra; Bondy, Jessica; Dziedzic, Amanda; Mora-Lagos, Barbara; Rivera-Alvarez, Gustavo; Ili-Gangas, Carmen; Brebi-Mieville, Priscilla; Westra, William; Koch, Wayne; Kang, Hyunseok; Marchionni, Luigi; Kim, Young; Sidransky, David

    2017-12-19

    Microbiome studies show altered microbiota in head and neck squamous cell carcinoma (HNSCC), both in terms of taxonomic composition and metabolic capacity. These studies utilized a traditional bioinformatics methodology, which allows for accurate taxonomic assignment down to the genus level, but cannot accurately resolve species level membership. We applied Resphera Insight, a high-resolution methodology for 16S rRNA taxonomic assignment that is able to provide species-level context in its assignments of 16S rRNA next generation sequencing (NGS) data. Resphera Insight applied to saliva samples from HNSCC patients and healthy controls led to the discovery that a subset of HNSCC saliva samples is significantly enriched with commensal species from the vaginal flora, including Lactobacillus gasseri/johnsonii (710x higher in saliva) and Lactobacillus vaginalis (52x higher in saliva). These species were not observed in normal saliva from Johns Hopkins patients, nor in 16S rRNA NGS saliva samples from the Human Microbiome Project (HMP). Interestingly, both species were only observed in saliva from Human Papilloma Virus (HPV) positive and HPV negative oropharyngeal cancer patients. We confirmed the representation of both species in HMP data obtained from mid-vagina (n=128) and vaginal introitus (n=121) samples. Resphera Insight also led to the discovery that Fusobacterium nucleatum , an oral cavity flora commensal bacterium linked to colon cancer, is enriched (600x higher) in saliva from a subset of HNSCC patients with advanced tumors stages. Together, these high-resolution analyses on 583 samples suggest a possible role for bacterial species in the therapeutic outcome of HPV positive and HPV negative HNSCC patients.

  6. Bacterial invasion of the uterus and oviducts in bovine pyometra

    DEFF Research Database (Denmark)

    Karstrup, C. C.; Pedersen, H. G.; Jensen, Tim Kåre

    2017-01-01

    of bacterial pathogenicity and development of lesions, have not been investigated. Bacterial invasion of the uterus and oviducts was studied in 21 cows diagnosed with pyometra at the time of slaughter by applying fluorescence in situ hybridization using probes targeting 16S ribosomal RNA of Fusobacterium...... necrophorum, Porphyromonas levii, Trueperella pyogenes and the overall bacterial domain Bacteria. Fusobacterium necrophorum and P. levii were found to invade the endometrium, especially if the endometrium was ulcerated, and penetrated deep into the lamina propria. These species co-localized within the tissue...... thus indicating a synergism. Trueperella pyogenes did not invade the uterine tissue. In addition to endometrial lesions, most cows with pyometra also had salpingitis but without significant bacterial invasion of the oviductal wall....

  7. Jugular thrombophlebitis complicating bacterial pharyngitis (Lemierre's syndrome)

    International Nuclear Information System (INIS)

    Sena, S.De; Rosenfeld, D.L.; Santos, S.; Keller, I.

    1996-01-01

    Lemierre's syndrome is a rare syndrome caused by Fusobacterium necrophorum, a gram-negative anaerobic organism that normally inhabits the oropharynx. The syndrome follows primary oropharyngeal infection and affects previously healthy adolescents in a characteristic manner with fatal results if left untreated. The authors present two cases seen at their institution and discuss the clinical and radiologic features of the syndrome, along with considerations for patient management. (orig.). With 5 figs

  8. Fusobacterial liver abscess: a case report and review of the literature.

    Science.gov (United States)

    Jayasimhan, Dilip; Wu, Linus; Huggan, Paul

    2017-06-20

    Fusobacteriae are facultative anaerobic gram-negative bacilli which cause a range of invasive infections, amongst which pyogenic liver abscesses are rare. We describe a case of Fusobacterium nucleatum liver abscess and review the relevant literature. A 51-year-old lady presented with a 4-day history of abdominal pain, diarrhoea, fever, rigors, and lethargy. Imaging revealed an abscess which was drained. Cultures of the blood and abscess aspirate grew Fusobacterium nucleatum and Prevotella pleuritidis respectively. She achieved full recovery following treatment. A MEDLINE search was undertaken using free-text and Medical Subject Headings (MeSH), keywords "Fusobacterium" and "Liver abscess". Non-English language reports and cases without confirmed growth of Fusobacterium species were excluded. Additional cases were identified by surveying the references of each report and by using the same keywords in a web-based search. Forty-eight cases were identified, 41 in men. The median age was 42.5, with an interquartile range of 33. F. nucleatum and F. necrophorum were in involved in 22 cases each, and 4 cases were not further speciated. Among cases of F. nucleatum liver abscess, nine were attributed to periodontal disease, four to lower gastrointestinal tract disease, one to Lemierre's Syndrome, and eight were considered cryptogenic. All patients treated made a full recovery. Antimicrobial treatment duration ranged from 2 weeks to 6 months with a median of 6 weeks. Fusobacterium nucleatum is an uncommon cause of liver abscess generally associated with good clinical outcomes with contemporary medical and surgical care.

  9. Lemierre's syndrome.

    LENUS (Irish Health Repository)

    O'Dwyer, D N

    2012-02-01

    Lemierre\\'s syndrome is a rare disease that results in an oropharyngeal infection, which precipitates an internal jugular vein thrombosis and metastatic infection. Fusobacterium necrophorum is an anaerobic Gram-negative bacillus and has been identified as the causative agent. We describe the case of a young girl whose presentation and diagnosis were confounded by a history of valvular heart disease. Infection of heart valves can produce many of the signs and symptoms associated with Lemierre\\'s syndrome. We describe the diagnosis, investigation and optimal management of this rare disorder.

  10. Pathogenesis and Treatment of Bovine Foot Rot.

    Science.gov (United States)

    Van Metre, David C

    2017-07-01

    Bovine foot rot (BFR) is an infectious disease of the interdigital skin and subcutaneous tissues of beef and dairy cattle that occurs under a variety of management and environmental settings. The anaerobic, gram-negative bacteria Fusobacterium necrophorum, Porphyromonas levii, and Prevotella intermedia are commonly isolated from lesions. A multitude of host, agent, and environmental factors contribute to the development of BFR. Initiation of systemic antimicrobial therapy early in the course of disease commonly leads to resolution. Delays in treatment may result in extension of infection into deeper bone, synovial structures, or ligamentous structures, and the prognosis for recovery is reduced. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Lester R. Dragstedt 1893-1975. Chronic ulcerative colitis. A summary of evidence implicating Bacterium necrophorum as an etiologic agent.

    Science.gov (United States)

    1988-08-01

    Lester Dragstedt was born in Anaconda, Montana, the son of Swedish immigrant parents. His entire college and professional education took place at the University of Chicago, where he received a B.S. degree in 1915, a master's degree in physiology in 1916, a Ph.D. in physiology in 1920, and the M.D. degree (from Rush) in 1921. His first academic appointment was as a physiologist at the State University of Iowa. In 1925 Dragstedt was recruited by Dallas B. Phemister to help design the new University Hospital research facilities on the campus of the University of Chicago. Following completion of this responsibility Phemister appointed Dragstedt as Associate Professor of Surgery, stating, "I can teach surgery to a physiologist; I am interested in teaching physiology to surgeons." In 1947 Dragstedt succeeded Phemister as chairman, a post he occupied until his retirement in 1959. Dragstedt was regarded as a skilled clinician as well as a dexterous and artistic surgeon. But he was particularly recognized for his contributions as physiologist-surgeon in the treatment of diseases of the pancreas, parathyroids, and especially diseases of the stomach. In 1943, he performed a transthoracic vagotomy on a patient with a duodenal ulcer who refused to accept the standard operation, subtotal gastrectomy. A lesser known but classical work of Dragstedt and his coworkers is reproduced here for this series. Dragstedt was the originator of the skin-grafted ileostomy in the treatment of ulcerative colitis. The author describes a complete "take" of the split-thickness graft in four patients, although he observed that the "resulting ileostomy looked somewhat like a penis." One can only surmise about the psychologic disability that would be produced. The stoma could, however, be fitted with an appliance that would minimize the risk of abdominal wall digestion. When reading the article and understanding the experimental studies proposing the possible causative organism of ulcerative colitis, one is impressed by Dragstedt's creative thinking. Dragstedt's renown as a basic scientist was illustrated by his election to the National Academy of Sciences. Following his Chicago retirement he became again a full-time physiologist with appointments as research professor in both the department of surgery and the department of physiology at the University of Florida College of Medicine. Active until the end, he died at his summer home on Elk Lake, Michigan on July 16, 1975.

  12. Potential bacterial core species associated with digital dermatitis in cattle herds identified by molecular profiling of interdigital skin samples

    DEFF Research Database (Denmark)

    Weiss Nielsen, Martin; Strube, Mikael Lenz; Isbrand, Anastasia

    2016-01-01

    of different molecular methods. Deep sequencing of the 16S rRNA gene variable regions V1–V2 showed that Treponema, Mycoplasma, Fusobacterium and Porphyromonas were the genera best differentiating the DD samples from the controls. Additional deep sequencing analysis of the most abundant genus, Treponema...... in the epidermal lesions and were present in only a subset of samples. RT-qPCR analysis showed that treponemes were also actively expressing a panel of virulence factors at the site of infection. Our results further support the hypothesis that species belonging to the genus Treponema are major pathogens of DD...... and also provide sufficient clues to motivate additional research into the role of M. fermentans, F. necrophorum and P. levii in the etiology of DD....

  13. Lemierre's Syndrome - A rare cause of disseminated sepsis requiring multi-organ support.

    Science.gov (United States)

    Rae, John; Misselbrook, Katie

    2017-11-01

    Lemierre's syndrome is a rare complication of acute pharyngitis characterised by septicaemia with infective thrombophlebitis of the internal jugular vein, most commonly due to Fusobacterium necrophorum . It characteristically affects healthy young adults causing persistent pyrexia and systemic sepsis presenting several days after an initial pharyngitis. Septic emboli seed via the bloodstream to distant sites including the lung, joints, skin, liver, spleen and brain. Prolonged antimicrobial therapy is required and admission to intensive care common. This once rare condition is increasing in incidence but awareness amongst clinicians is low. We present a classic case in a young man who developed multi-organ failure requiring intensive care support and describe the epidemiology, pathophysiology, microbiology, clinical features and management of the disease.

  14. A case of Lemierre's syndrome with septic shock and complicated parapneumonic effusions requiring intrapleural fibrinolysis

    Directory of Open Access Journals (Sweden)

    Daniel P. Croft

    2015-01-01

    Full Text Available Lemierre's syndrome is a septic thrombophlebitis of the internal jugular vein, which can lead to severe systemic illness. We report a case of an otherwise healthy 26-year-old man who suffered from pharyngitis followed by septic shock requiring intubation and vasopressor support from Fusobacterium necrophorum bacteremia. The septic emboli to his lungs caused complicated bilateral parapneumonic effusions, which recurred after initial drainage. He required bilateral chest tubes and intrapleural tPA to successfully drain his effusions. His fever curve and overall condition improved with the resolution of his effusions and after a 33-day hospitalization, he recovered without significant disability. The severity of his illness and difficult to manage complicated parapneumonic effusions were the unique facets of this case. Using an evidence-based approach of tPA and DNase for complicated parapneumonic effusions in Lemierre's syndrome can be safe and effective.

  15. Revisiting bovine pyometra-New insights into the disease using a culture-independent deep sequencing approach

    DEFF Research Database (Denmark)

    Knudsen, Lif Rødtness Vesterby; Karstrup, Cecilia Christensen; Pedersen, Hanne Gervi

    2015-01-01

    -independent studies have demonstrated that the bacterial diversity in most environments is underestimated in culture-based studies. Consequently, fastidious pyometra-associated pathogens may have been overlooked. Therefore, the primary purpose of this study was to investigate the diversity of bacteria in the uterus......The bacteria present in the uterus during pyometra have previously been studied using bacteriological culturing. These studies identified Fusobacterium necrophorum and Trueperella pyogenes as the major contributors to the pathogenesis of pyometra. However, an increasing number of culture...... of cows with pyometra by using culture-independent 16S rRNA PCR combined with next generation sequencing. We investigated the microbial composition in the uterus of 21 cows with pyometra, which were obtained from a Danish slaughterhouse. Similar to the observations from the culture studies...

  16. Lemierre's and Lemierre's-like syndromes in association with infectious mononucleosis.

    Science.gov (United States)

    Chacko, E M; Krilov, L R; Patten, W; Lee, P J

    2010-12-01

    This study aimed to review cases of Lemierre's and Lemierre's-like syndromes in paediatric patients, to examine a possible association with Epstein-Barr virus as a predisposing factor, and to assess the impact of this virus on the severity of illness. We performed a retrospective analysis of data from the in-patient database at Winthrop University Hospital, from January 2001 to October 2007. We reviewed clinical and laboratory findings as well as the outcome of infection in patients aged 21 years or less with a diagnosis of Lemierre's syndrome. An additional case of Lemierre's-like syndrome was also included. The illness severity and duration of in-patient management of those testing positive for heterophile antibody were then compared with the same parameters in patients who tested negative. Of the five patients diagnosed with Lemierre's syndrome, two had concomitant acute infection with Epstein-Barr virus. Additionally, a 19-year-old adolescent was admitted during this period with acute infectious mononucleosis, Fusobacterium necrophorum sepsis, sinusitis, frontal lobe abscess and ophthalmic vein thrombosis. The clinical presentation of all patients included fever, sore throat, and ear or neck pain. The duration of symptoms ranged from two days to three weeks prior to admission. The patients with acute Epstein-Barr virus infection had been diagnosed with infectious mononucleosis prior to admission, and tested positive for heterophile antibody. These patients subsequently underwent more extensive in-patient treatment, including intensive care management and ventilator support. The patients who tested negative for heterophile antibody experienced a milder course of illness, with a shorter duration of in-patient management. Two patients diagnosed with Lemierre's syndrome, and a third with Fusobacterium necrophorum sepsis, had coexisting acute Epstein-Barr virus infection. Patients who tested positive for heterophile antibody experienced a more severe course of

  17. Microbiological examination of infected dental root canals.

    Science.gov (United States)

    Gomes, B P F A; Pinheiro, E T; Gadê-Neto, C R; Sousa, E L R; Ferraz, C C R; Zaia, A A; Teixeira, F B; Souza-Filho, F J

    2004-04-01

    The aim of this study was to investigate the root canal microbiota of primary and secondary root-infected canals and the association of constituent species with specific endodontic signs and symptoms. Microbial samples were taken from 60 root canals, 41 with necrotic pulp tissues (primary infection) and 19 with failed endodontic treatment (secondary infection). Strict anaerobic techniques were used for serial dilution, plating, incubation and identification. A total of 224 cultivable isolates were recovered belonging to 56 different bacterial species. Individual root canals yielded a maximum of 10 bacterial species. Of the bacterial isolates, 70% were either strict anaerobes or microphilic. The anaerobes most frequently isolated were: Peptostreptococcus micros (35%), Fusobacterium necrophorum (23.3%), Fusobacterium nucleatum (11.7%), Prevotella intermedia/nigrescens (16.7%), Porphyromonas gingivalis (6.7%) and Porphyromonas endodontalis (5%). The root canal microflora of untreated teeth with apical periodontitis was found to be mixed, comprising gram-negative and gram-positive and mostly anaerobic microorganisms and usually containing more than 3 species per canal. On the other hand, facultative anaerobic and gram-positive bacteria predominated in canals with failed endodontic treatment, which harbored 1-2 species per canal. Suggested relationships were found between anaerobes, especially gram-negatives, and the presence or history of pain, tenderness to percussion and swelling (PEubacterium spp. (both Pspp. (Pspp. (Pspp. (Pspp. (Pspp. (Pspp. (Pspp. (Pspp., P. micros, F. necrophorum (P<0.05). Our findings indicate potential complex interactions of species resulting in characteristic clinical pictures which cannot be achieved by individual species alone. They also indicate that the microbiota of primary infected canals with apical periodontitis differs in number and in species from the secondary infected canals by using the culture technique.

  18. Lemierre’s syndrome: current perspectives on diagnosis and management

    Directory of Open Access Journals (Sweden)

    Johannesen KM

    2016-09-01

    Full Text Available Katrine M Johannesen,1 Uffe Bodtger1–3 1Department of Lung Medicine, Naestved Hospital, Naestved, 2Institute for Regional Health Research, University of Southern Denmark, Odense, 3Department of Pulmonology, Zealand University Hospital, Roskilde, Denmark Abstract: This is a systematic review of cases with Lemierre’s syndrome (LS in the past 5 years. LS is characterized by sepsis often evolving after a sore throat or tonsillitis and then complicated by various septic emboli and thrombosis of the internal jugular vein. Symptoms include sepsis, pain, and/or swelling in the throat or neck, as well as respiratory symptoms. Laboratory findings show elevated infectious parameters and radiological findings show thrombosis of the internal jugular vein and emboli in the lungs or other organs. The syndrome is often associated with an infection with Fusobacterium necrophorum. We found a total of 137 cases of LS, of which 47 were infected with F. necrophorum and others with Staphylococcus and Streptococcus. Complications of this rare but severe disease included osteomyelitis, meningitis, and acute respiratory distress syndrome. Mortality was extremely high in the pre-antibiotic era but has diminished with the advent of antibiotics. This review showed a mortality rate of only 2% of which none of the cases involved fusobacteria. Duration of treatment varied; a 4–6-week course of carbapenem or piperacillin/tazobactam in combination with metronidazole was optimum. Other treatment options included anticoagulants in 46% of cases, which is unwarrantedly high, as to date, no evidence of the positive effects of anticoagulants in LS exists. Only two cases had ligation of the internal jugular vein performed. This review confirms the rare, but severe aspects of LS. Mortality from LS in this day and age appears to be low, however the syndrome is difficult to recognize, and still requires the full attention of the clinician. Keywords: Lemierre’s syndrome

  19. Uterine bacterial flora in postpartum Danish Holstein dairy cows determined using DNA-based fingerprinting: correlation to uterine condition and calving management.

    Science.gov (United States)

    Elkjær, K; Ancker, M-L; Gustafsson, H; Friggens, N C; Waldmann, A; Mølbak, L; Callesen, H

    2013-04-01

    The overall aim of this study was to describe uterine bacterial flora during the postpartum period in Danish Holstein cows using the Terminal Restriction Fragment Length Polymorphism (T-RFLP) method. This method produces a pattern of nucleic acid fragments from the microorganisms present, reflecting the "fingerprint" of the actual microbial flora. As well as characterizing changes in flora with time from calving and between herds, data were examined for strong relations between uterine bacterial flora, calving management and uterine condition. In total 125 Holstein cows from five herds were included, and for each cow calving management was recorded. Cows were clinically examined on average 8 (range 0-19) and 28 (range 22-38) days after calving, and a uterine sample was taken for bacterial identification using T-RFLP. Milk samples were taken weekly for progesterone analysis. Bacteria were found in all cows at both examinations, and the flora was composed of many species, including species not traditionally reported to be present in the bovine uterus. The bacterial composition differed according to days from calving and herd. In all five herds Fusobacterium necrophorum, Pseudomonas/Acinetobacter and Bacteroides/Sphingobacterium/Prevotellaceae were among the most common at both examinations. In four herds there was a percentage decrease of F. necrophorum from first to second examination, and in all herds there was a percentage increase of Pseudomonas/Acinetobacter from first to second examination. No differences in bacterial flora were found between cows with different uterine scores, which were influenced by herd, calving difficulty and retained placenta. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Evolução Atípica de Angina de Plaut-Vincent em Criança: Relato de Caso/Atypical Evolution of Plaut-Vincent’s Angina in Child: A Case Report

    Directory of Open Access Journals (Sweden)

    Ananda Fortes Lima

    2014-09-01

    Full Text Available Introdução: A angina de Plaut-Vincent é uma afecção úlcero-necrótica causada pela simbiose de um bacilo fusiforme (Fusobacterium necrophorum e um espirilo (Borrelia vincenti. Ambos são agentes saprófitas habituais da cavidade oral, porém, quando associados, determinam o caráter patogênico da doença. Casuística: Trata-se do caso de uma paciente do sexo feminino, 4 anos de idade, que deu entrada no pronto socorro com choque séptico descompensado não responsivo a volume com necessidade de drogas vasoativas (DVA, além de má conservação dentária, e amigdalas necrosadas. Em 48h evoluiu com piora hemodinâmica refratária a DVA e introdução de hidrocortisona, com crises convulsivas e hemiplegia à esquerda. Foi solicitada TC crânio a qual evidenciou AVC isquêmico. Discussão: Ocorre geralmente em condições de desnutrição e má higiene oral, tendo sua maior incidência entre os adultos-jovens e adolescentes. Cursa com necrose das amigdalas, halitose intensa e disfagia. Conclusão: Considerando que a Angina de Plaut-Vincent acomete principalmente adultos jovens, torna-se de grande relevância relatar o caso de uma paciente pediátrica, que evoluiu de forma incomum, apresentando lesões isquêmicas cerebrovasculares. Introduction: Plaut–Vincent’s angina is an ulcer-necrotic disease caused by the symbiosis of a fusiform bacillus (Fusobacterium necrophorum and spirillum (Borrelia vincenti. Both are common saprophytic agents of the oral cavity, but when combined determine the pathogenic of the disease. Case report: We describe the case of a female patient, 4 years old, which gave entered the emergency room with decompensated septic shock unresponsive to volume in the need of vasoactive drugs (DVA, moreover poor dental conservation and necrotic tonsils. After 48h evolved with hemodynamic deterioration refractory to VAD and insertion of hydrocortisone, with convulsion and left hemiplegia. A cranial CT was requested and revealed

  1. Spontaneously draining acute otitis media in children: an observational study of clinical findings, microbiology and clinical course.

    Science.gov (United States)

    Neumark, Thomas; Ekblom, Maria; Brudin, Lars; Groth, Anita; Eliasson, Ingvar; Mölstad, Sigvard; Petersson, Ann-Cathrine; Törngren, Annika

    2011-12-01

    To study the outcome of acute otitis media (AOM) with otorrhoea in children managed initially without antibiotics, in relation to bacterial and clinical findings, and to identify those who may benefit from antibiotics. Otherwise healthy, not otitis prone children aged 2-16 y, presenting with AOM with spontaneous otorrhoea, were recruited from primary care and followed at selected ear, nose and throat (ENT) clinics. Specimens for bacterial investigations were obtained; symptoms were registered on a daily basis. The main outcomes measured were the frequency of children treated with antibiotics due to persisting AOM within 9 days in relation to clinical and bacteriological findings, and new AOM within 3 months. Twelve of 71 children who completed the trial received antibiotics during the first 9 days due to lack of improvement. One received antibiotics after 16 days due to relapsing AOM and 6 received antibiotics after 30 days due to new AOM. At 2-4 days following inclusion, over 70% of children showed normalized eardrum status and markedly reduced secretion. Alloiococcus otitidis was found in 23 samples, Streptococcus pneumoniae in 12, Streptococcus pyogenes in 6, and Fusobacterium nucleatum in 5. Mycoplasma pneumoniae, Chlamydia pneumoniae, and Fusobacterium necrophorum were not detected. Antibiotics were prescribed more extensively to children with a pulsating eardrum and abundant purulent secretion. All children with S. pyogenes received antibiotics, whereas children with only A. otitidis did not. The results suggest that antibiotics are indicated in AOM with otorrhoea and the presence of abundant purulent secretion, a pulsating eardrum, or the presence of S. pyogenes. The presence of only A. otitidis was not associated with a more prolonged course or the need for antibiotics.

  2. Molecular Mapping to Species Level of the Tonsillar Crypt Microbiota Associated with Health and Recurrent Tonsillitis

    Science.gov (United States)

    Jensen, Anders; Fagö-Olsen, Helena; Sørensen, Christian Hjort; Kilian, Mogens

    2013-01-01

    The human palatine tonsils, which belong to the central antigen handling sites of the mucosal immune system, are frequently affected by acute and recurrent infections. This study compared the microbiota of the tonsillar crypts in children and adults affected by recurrent tonsillitis with that of healthy adults and children with tonsillar hyperplasia. An in-depth 16S rRNA gene based pyrosequencing approach combined with a novel strategy that included phylogenetic analysis and detection of species-specific sequence signatures enabled identification of the major part of the microbiota to species level. A complex microbiota consisting of between 42 and 110 taxa was demonstrated in both children and adults. This included a core microbiome of 12 abundant genera found in all samples regardless of age and health status. Yet, Haemophilus influenzae, Neisseria species, and Streptococcus pneumoniae were almost exclusively detected in children. In contrast, Streptococcus pseudopneumoniae was present in all samples. Obligate anaerobes like Porphyromonas, Prevotella, and Fusobacterium were abundantly present in children, but the species diversity of Porphyromonas and Prevotella was larger in adults and included species that are considered putative pathogens in periodontal diseases, i.e. Porphyromonas gingivalis, Porphyromonas endodontalis, and Tannerella forsythia. Unifrac analysis showed that recurrent tonsillitis is associated with a shift in the microbiota of the tonsillar crypts. Fusobacterium necrophorum, Streptococcus intermedius and Prevotella melaninogenica/histicola were associated with recurrent tonsillitis in adults, whereas species traditionally associated with acute tonsillitis like pyogenic streptococci and Staphylococcus aureus were scarce. The findings suggest that recurrent tonsillitis is a polymicrobial infection in which interactions within consortia of taxa play an etiologic role. The study contributes to the human microbiome data, to the understanding of the

  3. Molecular mapping to species level of the tonsillar crypt microbiota associated with health and recurrent tonsillitis.

    Directory of Open Access Journals (Sweden)

    Anders Jensen

    Full Text Available The human palatine tonsils, which belong to the central antigen handling sites of the mucosal immune system, are frequently affected by acute and recurrent infections. This study compared the microbiota of the tonsillar crypts in children and adults affected by recurrent tonsillitis with that of healthy adults and children with tonsillar hyperplasia. An in-depth 16S rRNA gene based pyrosequencing approach combined with a novel strategy that included phylogenetic analysis and detection of species-specific sequence signatures enabled identification of the major part of the microbiota to species level. A complex microbiota consisting of between 42 and 110 taxa was demonstrated in both children and adults. This included a core microbiome of 12 abundant genera found in all samples regardless of age and health status. Yet, Haemophilus influenzae, Neisseria species, and Streptococcus pneumoniae were almost exclusively detected in children. In contrast, Streptococcus pseudopneumoniae was present in all samples. Obligate anaerobes like Porphyromonas, Prevotella, and Fusobacterium were abundantly present in children, but the species diversity of Porphyromonas and Prevotella was larger in adults and included species that are considered putative pathogens in periodontal diseases, i.e. Porphyromonas gingivalis, Porphyromonas endodontalis, and Tannerella forsythia. Unifrac analysis showed that recurrent tonsillitis is associated with a shift in the microbiota of the tonsillar crypts. Fusobacterium necrophorum, Streptococcus intermedius and Prevotella melaninogenica/histicola were associated with recurrent tonsillitis in adults, whereas species traditionally associated with acute tonsillitis like pyogenic streptococci and Staphylococcus aureus were scarce. The findings suggest that recurrent tonsillitis is a polymicrobial infection in which interactions within consortia of taxa play an etiologic role. The study contributes to the human microbiome data, to the

  4. The Anticomplementary Activity of ’Fusobacterium polymorphum’ in Normal and C-4 Deficient Sources of Guinea Pig Complement.

    Science.gov (United States)

    1977-01-12

    A complement consumption assay was used to show that the anticomplementary activity of a cell wall preparation from F. polymorphum in guinea pig complement...tests with C𔃾-deficient guinea pig sera confirmed that F. polymorphum cell walls were capable of generating alternate complement pathway activity in guinea pig sera.

  5. Differential Susceptibility of Bacteria to Mouse Paneth Cell a-Defensins under Anaerobic Conditions

    Directory of Open Access Journals (Sweden)

    Jennifer R. Mastroianni

    2014-10-01

    Full Text Available Small intestinal Paneth cells secrete a-defensin peptides, termed cryptdins (Crps in mice, into the intestinal lumen, where they confer immunity to oral infections and define the composition of the ileal microbiota. In these studies, facultative bacteria maintained under aerobic or anaerobic conditions displayed differential sensitivities to mouse a-defensins under in vitro assay conditions. Regardless of oxygenation, Crps 2 and 3 had robust and similar bactericidal activities against S. typhimurium and S. flexneri, but Crp4 activity against S. flexneri was attenuated in the absence of oxygen. Anaerobic bacteria varied in their susceptibility to Crps 2-4, with Crp4 showing less activity than Crps 2 and 3 against Enterococcus faecalis, and Bacteroides fragilis in anaerobic assays, but Fusobacterium necrophorum was killed only by Crp4 and not by Crps 2 and 3. The influence of anaerobiosis in modulating Crp bactericidal activities in vitro suggests that a-defensin effects on the enteric microbiota may be subject to regulation by local oxygen tension.

  6. Dyskospondylitis and paravertebral abscesses in a calf

    International Nuclear Information System (INIS)

    Testoni, S.

    2006-01-01

    A case of progressive spastic paraparesis due to a dyskospondylitis at the level of T9-T10 is described in a four-month old Holstein female calf. The calf was recumbent, but bright, alert and willing to suckle. Despite repeated attempts, the calf was not able to assume the sternal recumbency. The radiological findings were decisive for the in life diagnosis. Spinal radiography of the thoraco-lumbar region revealed lysis and collapse of T9 and T10 vertebral bodies; irregular proliferative new bone was evident. Lumbo-sacral myelography showed a narrowing and dorsal displacement of the ventral contrast column at the same spinal level, indicating a severe ventral extradural compression of the spinal cord. At the level of the thoracic cavity, a 20 x 15 cm diameter opacity extending ventrally to T8-T13 and caudo-dorsally to the heart was also evident. At gross necroscopy, two approximately 15 cm diameter encapsulated paravertebral abscesses were evident in the thoracis cavity just below the spinal column. A pure culture of Fusobacterium necrophorum was obtained from them. A saggital section of the spine showed an erosive suppurative process of T9 and T10 vertebral bodies that provoked the compression of the thoracic tract of the spinal cord [it

  7. Antimicrobial activity of some Pacific Northwest woods against anaerobic bacteria and yeast.

    Science.gov (United States)

    Johnston, W H; Karchesy, J J; Constantine, G H; Craig, A M

    2001-11-01

    Extracts of woods commonly used for animal bedding were tested for antimicrobial activity. Essential oils from Alaska cedar (Chamaecyparis nootkatensis), western juniper (Juniperus occidentalis) and old growth Douglas fir (Pseudotsuga menziesii) as well as methanol extracts of wood from these trees plus western red cedar (Thuja plicata) and ponderosa pine (Pinus ponderosa) were tested for antimicrobial activity against anaerobic bacteria and yeast. The test microbes included Fusobacterium necrophorum, Clostridium perfringens, Actinomyces bovis and Candida albicans which are common to foot diseases and other infections in animals. The essential oils and methanol extracts were tested using a standardized broth assay. Only extracts of Alaska cedar and western juniper showed significant antimicrobial activity against each of the microbes tested. The essential oil of Douglas fir did show antimicrobial activity against A. bovis at the concentrations tested. The methanol extracts of the heartwood of Douglas fir and the sapwood of ponderosa pine showed no antimicrobial activity. The major chemical components of western juniper (cedrol and alpha- and beta-cedrene) and Alaska cedar (nootkatin) were also tested. In western juniper, alpha- and beta-cedrene were found to be active components. Nootkatin showed activity only against C. albicans. The inhibitory activity in Alaska cedar oil was high enough to justify further efforts to define the other chemical components responsible for the antimicrobial activity. Copyright 2001 John Wiley & Sons, Ltd.

  8. Presence of bacteria in the endometrium and placentomes of pregnant cows.

    Science.gov (United States)

    Karstrup, Cecilia Christensen; Klitgaard, Kirstine; Jensen, Tim Kåre; Agerholm, Jørgen Steen; Pedersen, Hanne Gervi

    2017-09-01

    Bacterial invasion of the bovine uterus during the postpartum period occurs in most cows, but the general consensus is that these bacteria are eliminated before the next pregnancy. The pregnant uterus has therefore hitherto been considered a sterile environment, but this assumption has now been challenged by recent studies in humans, which indicate that bacteria can be present in the placenta of term pregnancies without causing abortion. The aim of the present study was therefore to investigate whether bacteria are present in the uterus of pregnant cows. Specimens were taken from the inter-caruncular endometrium and from placentomes of slaughtered pregnant cows (n = 43) and subjected to histology, fluorescence in situ hybridization and massive parallel sequencing. Bacteria were observed in the tissue from 90.7% (39/43) of the cows by fluorescence in situ hybridization. Fusobacterium necrophorum, Porphyromonas levii and Trueperella pyogenes were located within the endometrium, on the endometrial surface and in the caruncular stroma, but their presence was not associated with inflammation. Data from massive parallel sequencing of the 16S rRNA gene from a subset of 15 cows indicated that the most abundant bacteria were the families Porphyromonadaceae, followed by Ruminococcaceae and Lachnospiraceae. Our results indicate that the bovine uterus is not a sterile environment during pregnancy as previously assumed and that a cow can carry a pregnancy despite the presence of a few potentially pathogenic bacteria in the uterus. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Adherence of bacteria to mucus collected from different parts of the reproductive tract of heifers and cows.

    Science.gov (United States)

    Styková, E; Nemcová, R; Valocký, I; Novotný, F; Guba, P

    2013-11-01

    In the present study, we examined the adherence of indigenous vaginal bacteria, probiotic strains, and metritis pathogens to mucus collected from different parts of the reproductive tracts of heifers and cows and compared their adherence with the bacterial adherence to mucus collected from the stomach and large intestine of pigs. Most of the vaginal strains adhered to mucus collected from different parts of the reproductive tract and strongly adhered to gastric mucus, with the exception of Lactobacillus buchneri 24S8. Only Lactobacillus mucosae 29S8, Enterococcus faecium E21, and E. faecium EAC adhered to colonic mucus. Probiotic strains adhered strongly to mucus collected from the reproductive tract and gastric mucus but did not adhere to colonic mucus. Pathogenic strains were adherent to vaginal, uterine horn, and gastric mucus, except Escherichia coli O8:K88ab:H9 (65), Fusobacterium necrophorum, and Gardnerella vaginalis, which adhered to uterine cervix mucus. Only Kocuria kristinae and G. vaginalis adhered to uterine body mucus; E. coli O149:K88ac (EC) adhered to colonic mucus. The strains did not exhibit host specificity but rather strain specificity. The ability to adhere to mucus was a characteristic unique to each strain. To our knowledge, this is the first report regarding in vitro adherence of GRAS (Generally Regarded As Safe) lactobacilli isolated from different sources to mucus collected from different parts of the reproductive tract.

  10. Bovine vaginal strain Kocuria kristinae and its characterization.

    Science.gov (United States)

    Styková, Eva; Nemcová, Radomíra; Gancarčíková, Soňa; Valocký, Igor; Lauková, Andrea

    2016-05-01

    Kocuria spp. are widely distributed in nature. They are Gram-positive, coagulase-negative, coccoid bacteria belonging to the family Micrococcaceae, suborder Micrococcineae, order Actinomycetales, class Actinobacteria. In general, limited knowledge exists concerning the properties associated with the representants of the genus Kocuria, Kocuria kristinae as well. Following our previous results, K. kristinae Kk2014 Biocenol(™) (CCM 8628) was isolated from vagina of a healthy cow. Its taxonomical allottation was confirmed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) identification system and phenotypic characteristics. Kk2014 strain showed strong adherence capability to the vaginal mucus, produced organic acids which can play a role in prevention of unsuitable contamination, and showed in vitro antagonistic/antimicrobial activity against strains Arcanobacterium pyogenes CCM 5753, Fusobacterium necrophorum CCM 5982, Streptococcus equi subsp. zooepidemicus CCM 7316, and Gardnerella vaginalis CCM 6221. Antimicrobial activity ranged from 100 to 200 AU/mL, up to 32 mm in size, respectively.

  11. Oro-facial gangrene (noma/cancrum oris): pathogenetic mechanisms.

    Science.gov (United States)

    Enwonwu, C O; Falkler, W A; Idigbe, E O

    2000-01-01

    Cancrum oris (Noma) is a devastating infectious disease which destroys the soft and hard tissues of the oral and para-oral structures. The dehumanizing oro-facial gangrenous lesion affects predominantly children ages 2 to 16 years, particularly in sub-Saharan Africa, where the estimated frequency in some communities varies from 1 to 7 cases per 1000 population. The risk factors are poverty, malnutrition, poor oral hygiene, residential proximity to livestock in unsanitary environments, and infectious diseases, particularly measles and those due to the herpesviridae. Infections and malnutrition impair the immune system, and this is the common denominator for the occurrence of noma. Acute necrotizing gingivitis (ANG) and oral herpetic ulcers are considered the antecedent lesions, and ongoing studies suggest that the rapid progression of these precursor lesions to noma requires infection by a consortium of micro-organisms, with Fusobacterium necrophorum (Fn) and Prevotella intermedia (Pi) as the suspected key players. Additional to production of a growth-stimulating factor for Pi, Fn displays a classic endotoxin, a dermonecrotic toxin, a cytoplasmic toxin, and a hemolysin. Without appropriate treatment, the mortality rate from noma is 70-90%. Survivors suffer the two-fold afflictions of oro-facial mutilation and functional impairment, which require a time-consuming, financially prohibitive surgical reconstruction.

  12. The oral microbial community of gingivitis and lumpy jaw in captive macropods.

    Science.gov (United States)

    Antiabong, John F; Boardman, Wayne; Moore, Robert B; Brown, Melissa H; Ball, Andrew S

    2013-12-01

    Gingivitis and lumpy jaw are diseases of polymicrobial aetiology. Although Fusobacterium necrophorum has been associated with these diseases in macropods, little is known about other organisms associated with these diseases in this animal species. PCR-DGGE analysis revealed the potential pathogens associated with gingivitis and lumpy jaw in macropods. PCR-DGGE profile comparison between the healthy and disease groups indicated a shift in the oral bacterial community structures with similarity coefficients of 48% and 35% for gingivitis and lumpy jaw respectively. Moreover, gingivitis was associated with increase in bacterial diversity (Shannon index = 2.87; PL curve = 45%) while lumpy jaw resulted in a decline in bacterial diversity (Shannon index = 2.47; PL curve = 74%). This study suggest that the establishment of gingivitis and lumpy jaw diseases follows the ecological plaque hypothesis. This forms the basis for an expanded investigation in an epidemiological scale and suggests the need for the appropriate choice of antimicrobial agent(s) and for the effective management and control of polymicrobial diseases. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Lemierre syndrome: two cases and a review.

    Science.gov (United States)

    Syed, Mohammed Iqbal; Baring, David; Addidle, Michael; Murray, Craig; Adams, Calum

    2007-09-01

    Lemierre syndrome is usually caused by an acute oropharyngeal infection in previously healthy young adults, resulting in thrombophlebitis of the internal jugular vein, leading to metastatic septic embolization and bacteraemia. The usual organism is Fusobacterium necrophorum. Lemierre syndrome, not so long ago labeled as the "forgotten disease," is on the rise. Today with increasing antibiotic-resistant organisms, and decreasing awareness of the syndrome, subsequent re-emergence of this "forgotten disease" is becoming more common in clinical settings. Lemierre syndrome has significant morbidity. Cranial nerve complications associated with the condition have been increasingly diagnosed in the last few years. Looking back at literature on Lemierre syndrome, there have been review articles in medical and microbiology journals but rarely in otolaryngology journals. By presenting our cases we demonstrate the diverse presentations and severity of the illness. A review of the literature and a case report on two cases seen in our institution in the last year are presented. Each of these had varied presentations and neurologic complications-one developed 9th to 12th cranial nerve palsies and Horner syndrome, which have not been described in previous literature, and the other developed polyneuropathy and a frontal lobe infarct among other multisystem complications. Diagnosis of Lemierre syndrome is not always straightforward as clinical features are variable and blood cultures are often negative. Awareness of the syndrome and a high degree of suspicion are needed.

  14. The role of DNA amplification and cultural growth in complicated acute appendicitis

    Directory of Open Access Journals (Sweden)

    Francesca Tocchioni

    2016-09-01

    Full Text Available Bacterial growth of peritoneal fluid specimens obtained during surgical procedures for acute appendicitis may be useful to optimize further antibiotic therapy in complicated cases. DNA amplification represents a fast technique to detect microbial sequences. We aimed to compare the potential of DNA amplification versus traditional bacterial growth culture highlighting advantages and drawbacks in a surgical setting. Peritoneal fluid specimens were collected during surgery from 36 children who underwent appendectomy between May and December 2012. Real-time polymerase chain reaction (RT-PCR and cultures were performed on each sample. RT-PCR showed an amplification of 16S in 18/36 samples, Escherichia coli (in 7 cases, Pseudomonas aeruginosa (3, Fusobacterium necrophorum (3, Adenovirus (2, E.coli (1, Klebsiella pneumoniae (1, Serratia marcescens/Enterobacter cloacae (1. Bacterial growth was instead observed only in four patients (3 E.coli and 1 P.aeruginosa and Bacteroides ovatus. Preoperative C-reactive protein and inflammation degree, the most reliable indicators of bacterial translocation, were elevated as expected. DNA amplification was a quick and useful method to detect pathogens and it was even more valuable in detecting aggressive pathogens such as anaerobes, difficult to preserve in biological cultures; its drawbacks were the lack of biological growths and of antibiograms. In our pilot study RT-PCR and cultures did not influence the way patients were treated.

  15. 21 CFR 520.2261b - Sulfamethazine powder.

    Science.gov (United States)

    2010-04-01

    ... and bovine respiratory disease complex (shipping fever complex) (Pasteurella spp.), colibacillosis... (F. necrophorum), acute mastitis (Streptococcus spp.), and acute metritis (Streptococcus spp.) (iii...

  16. Lemierre’s syndrome following perianal abscess: A case report

    Directory of Open Access Journals (Sweden)

    Qusai Aljarrah

    Full Text Available Introduction: Lemierre’s syndrome (LS is a rare and life-threatening condition characterized by suppurative thrombophlebitis of the internal jugular vein (IJV, and a history of head and neck (H&N sepsis. LS is usually caused by Fusobacterium necrophorum, which is part of the normal flora in the oro-pharynx, and the digestive and urogenital tracts. We here report the first case of LS following perianal sepsis. Presentation of case: A 60-year-old man with a painful left neck swelling, dysphagia and worsening sepsis was referred from a peripheral unit where he had an incision and drainage of a perianal abscess a week earlier. Urgent Doppler ultrasound and computed tomographic scans demonstrated suppurative thrombophlebitis of the left IJV, and the patient was subsequently commenced on intravenous Piperacillin/Tazobactam and heparin. The symptoms gradually improved, and the patient was eventually discharged on the 10th day. Discussion: Vigilant examination of the H&N region searching for a primary source is paramount, but LS following infections in the gastrointestinal or uro-genital tracts has also been described. A high index of suspicion is required for diagnosis, especially in patients with unresolving pharyngitis with a unilateral neck swelling, and septicaemia. Early resuscitation and treatment with broad-spectrum parenteral antimicrobials are important for favourable outcome. Conclusion: LS is well known to specialists in the H&N region, but other disciplines like general surgery, urology, or obstetrics and gynaecology might also rarely encounter the disease. We present a case of LS complicating a perianal abscess that was successfully treated with good outcome. Keywords: Lemierre’s syndrome, Thrombophlebitis, Internal jugular vein, Perianal abscess, Sepsis

  17. The influence of incubation time, sample preparation and exposure to oxygen on the quality of the MALDI-TOF MS spectrum of anaerobic bacteria.

    Science.gov (United States)

    Veloo, A C M; Elgersma, P E; Friedrich, A W; Nagy, E; van Winkelhoff, A J

    2014-12-01

    With matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), bacteria can be identified quickly and reliably. This accounts especially for anaerobic bacteria. Because growth rate and oxygen sensitivity differ among anaerobic bacteria, we aimed to study the influence of incubation time, exposure to oxygen and sample preparation on the quality of the spectrum using the Bruker system. Also, reproducibility and inter-examiner variability were determined. Twenty-six anaerobic species, representing 17 genera, were selected based on gram-stain characteristics, growth rate and colony morphology. Inter-examiner variation showed that experience in the preparation of the targets can be a significant variable. The influence of incubation time was determined between 24 and 96 h of incubation. Reliable species identification was obtained after 48 h of incubation for gram-negative anaerobes and after 72 h for gram-positive anaerobes. Exposure of the cultures to oxygen did not influence the results of the MALDI-TOF MS identifications of all tested gram-positive species. Fusobacterium necrophorum and Prevotella intermedia could not be identified after >24 h and 48 h of exposure to oxygen, respectively. Other tested gram-negative bacteria could be identified after 48 h of exposure to oxygen. Most of the tested species could be identified using the direct spotting method. Bifidobacterium longum and Finegoldia magna needed on-target extraction with 70% formic acid in order to obtain reliable species identification and Peptoniphilus ivorii a full extraction. Spectrum quality was influenced by the amount of bacteria spotted on the target, the homogeneity of the smear and the experience of the examiner. © 2014 The Authors Clinical Microbiology and Infection © 2014 European Society of Clinical Microbiology and Infectious Diseases.

  18. The effect of cyclosporin-A on the oral microflora at gingival sulcus of the ferret.

    Science.gov (United States)

    Fischer, R G; Edwardsson, S; Klinge, B; Attström, R

    1996-09-01

    The effect of cyclosporin-A (CyA) on the dentogingival flora of ferrets with healthy and experimentally induced periodontal breakdown was studied. Five animals were given 10 mg/kg/d CyA. At the start of the experiments (day 0), ligatures were placed around 4 teeth in the right upper and lower jaws; corresponding contralateral teeth on the left side served as control. On days 0 and 28 (end of the experiment), microbiological samples were collected from the gingival sulcus of the experimental and the control teeth and from closely located gingival mucosa membrane. The samples were subjected to viable counts and to darkfield microscopic analyses. On day 0, facultative anaerobic rods, mainly Pasteurella spp, Alcaligenes spp, Corynebacterium spp. and Rothia spp dominated in the viable counts. No anaerobic bacteria were detected in the viable counts. On day 28 spirochetes increased in the experimental gingival sulcus samples and anaerobic bacteria appeared in most of the samples and constituted 40-60% of the total cultivable flora; Fusobacterium necrophorum and Eubacterium spp. predominated in the samples from the experimental sites. The results of the present study were compared with those of our previous investigation of ferrets not medicated with cyclosporin but also subject to experimental ligature periodontitis. Eubacterium spp. were absent in the animals not treated with cyclosporin, while this species was frequently present in the immunosuppressed ferrets. The results indicate that the presence of the large numbers of gram negative rods and of anaerobic bacteria may have enhanced the inflammatory process and further provoked the gingival overgrowth observed.

  19. Periodontal-disease-associated biofilm: A reservoir for pathogens of medical importance.

    Science.gov (United States)

    Vieira Colombo, Ana Paula; Magalhães, Clarissa Bichara; Hartenbach, Fátima Aparecida Rocha Resende; Martins do Souto, Renata; Maciel da Silva-Boghossian, Carina

    2016-05-01

    The ecological diversity of the periodontal microenvironment may provide suitable conditions for the colonization of species not usually considered members of the oral microbiota. In this investigation, we aimed to determine the prevalence and levels of pathogenic species of medical relevance in the microbiota of individuals with distinct periodontal clinical status. Subgingival biofilm was obtained from patients with periodontal health (H, n = 81), gingivitis (G, n = 55), generalized aggressive (AgP, n = 36) or chronic periodontitis (CP, n = 98), and analyzed for 39 microbial taxa using a checkerboard DNA-DNA hybridization technique. Microbial differences among groups, as well as associations between clinical and microbiological parameters were sought by non-parametric and univariate correlation tests. Neisseria spp., Peptostreptococus anaerobius, Candida albicans, enterobacteria, Pseudomonas aeruginosa, Eubacterium saphenum, Clostridium difficile and Olsenella uli were detected in high mean prevalence and counts in the subgingival microbiota of the study population. Species that were more related to periodontal inflammation and tissue destruction at the patient and site levels included enterobacteria, C. albicans, Neisseria spp., P. aeruginosa, O. uli, Hafnia alvei, Serratia marcescens and Filifactor alocis (p < 0.05). In contrast, Fusobacterium necrophorum, Lactobacillus acidophilus, Staphylococcus aureus and Streptococcus pneumoniae were associated with periodontal health (p < 0.05). Pathogenic species of medical importance may be detected in high prevalence and levels in the periodontal microbiota. Regardless of their role in periodontal health or disease, the periodontal biofilm may be a source for dissemination and development of systemic infections by these pathogenic microorganisms. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Synergic phototoxic effect of visible light or Gallium-Arsenide laser in the presence of different photo-sensitizers on Porphyromonas gingivalis and Fusobacterium nucleatum

    Directory of Open Access Journals (Sweden)

    Habibollah Ghanbari

    2015-01-01

    Conclusion: Within the limitations of this study, the synergic phototoxic effect of visible light in combination with each of the photosensitizers on P. gingivalis and F. nucleatum. However, the synergic phototoxic effect of laser exposure and hydrogen peroxide and curcumin as photosensitizers on F. nucleatum was not shown.

  1. Lameness in feedlot cattle.

    Science.gov (United States)

    Stokka, G L; Lechtenberg, K; Edwards, T; MacGregor, S; Voss, K; Griffin, D; Grotelueschen, D M; Smith, R A; Perino, L J

    2001-03-01

    This article examines the various causes of lameness in feedlot cattle, with an emphasis on clinical signs, treatment, and prevention. Specific conditions are discussed, including interdigital necrobacillosis, laminitis, feedlot injuries, and feedlot lameness associated with Mycoplasma bovis. Immune management of the foot is also reviewed.

  2. Lemierre\\'s syndrome - the uncommon cold | Such | SA Journal of ...

    African Journals Online (AJOL)

    Lemierre's syndrome (postanginal sepsis/necrobacillosis) is a rare clinical entity characterised by the triad of acute oropharyngeal infection, with secondary thrombophlebitis of the internal jugular vein and metastatic septic emboli.A high degree of clinical suspicion is needed to diagnose the syndrome and the radiologist's ...

  3. 77 FR 735 - New Animal Drugs; Cephalosporin Drugs; Extralabel Animal Drug Use; Order of Prohibition

    Science.gov (United States)

    2012-01-06

    ... intramammary infusion for the treatment of lactating cows having bovine mastitis caused by susceptible strains..., sheep, and goats; (2) the treatment of acute bovine interdigital necrobacillosis (foot rot) and acute bovine metritis; (3) the control of bovine respiratory disease; and (4) the control of early mortality...

  4. Peritonsillar abscess: clinical aspects of microbiology, risk factors, and the association with parapharyngeal abscess.

    Science.gov (United States)

    Klug, Tejs Ehlers

    2017-03-01

    transportation of appropriate specimens, choice of methodology for detection and quantification of microorganisms, current or recent antibiotic treatment of patients, potential shift in significant pathogens during the course of infection, and factors associated with increased risk of PTA development.  The trend towards de-escalated surgical intervention and increasing reliance on antibiotic treatment, require studies defining the significant pathogens in PTA in order to determine optimal antibiotic regimens. Complications secondary to PTA may be avoided or better controlled with improved knowledge concerning the significant pathogens in PTA. Furthermore, identification of pathogens other than GAS, may lead the way for earlier bacterial diagnosis and timely intervention before abscess formation in sore throat patients. The identification and quantification of risk factors for PTA development constitutes another approach to reduce the incidence of PTA. As clinicians, we noticed that FN was recovered from PTA patients with increasing frequency and that patients infected with Fusobacterium necrophorum (FN) seemed to be more severely affected than patients infected with other bacteria. Furthermore, we occationally observed concomitant PPA in addition to a PTA, which made us hypothesize that PPA and PTA is often closely related and may share significant pathogens. Hence, our aims were: 1. To explore the microbiology of PTA with a special attention to Fusobacterium necrophorum (FN). 2. To elucidate whether smoking, age, gender, and seasons are risk factors for the development of PTA. 3. To characterize patients with PPA, explore the relationship between PPA and PTA, identify the pathogens associated with PPA, and review our management of PPA. In a retrospective study on all 847 PTA patients admitted to the ENT department at Aarhus University Hospital (AUH) from 2001 to 2006, we found that FN was the most prevalent (23%) bacterial strain in pus specimens. FN-positive patients

  5. Lemierre’s syndrome and right adnexal abscess; A case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Malak Al-Hakeem

    2014-09-01

    Full Text Available Lemierre’s syndrome is an extremely rare but a completely curable condition. A high index of suspicion is needed for early diagnosis and proper treatment. We report a 20 year old virgin Caucasian lady presented with sore throat, fever, neck pain, nausea and vomiting followed by acute abdomen. Her laboratory investigations revealed white blood count 13,300/mm3, C-reactive protein 332 mg/L. Abdominal computed tomography scan showed pelvic abscess 6.2 x 6.1 cm mainly involving the right ovary. Intra-abdominal swab showed growth of Fusobactrerium necrophorum. Right adnexectomy and proper antibiotic was carried out. High index of doubt is the most important key in diagnosing this fatal syndrome. In our case the diagnosis of the syndrome was made only after the isolation of F. necrophorum from the intra-abdominal swab culture.J Microbiol Infect Dis 2014; 4(3: 114-117

  6. Therapeutic effect of Lianbeijuqin (a Chinese herbal cocktail) on ...

    African Journals Online (AJOL)

    Additionally, the antibacterial activity of LBJQ against Porphyromonas gingivalis, Prevotella intermedius, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Streptococcus sanguis and Streptococcus mutans were evaluated using minimal inhibitory concentration (MIC) and minimal bactericidal concentration ...

  7. Antibiotic susceptibility profiles of anaerobic pathogens in The Netherlands

    NARCIS (Netherlands)

    Veloo, A. C. M.; van Winkelhoff, A. J.

    The antibiotic susceptibility profile of the Bacteroides fragilis group, Gram-positive anaerobic cocci (GPAC), Fusobacterium spp., Prevotella spp., Veillonella spp. and Bilophila wadsworthia for amoxicillin, amoxicillin-clavulanic acid, clindamycin and metronidazole was determined. Human clinical

  8. Molecular mapping to species level of the tonsillar crypt microbiota associated with health and recurrent tonsillitis

    DEFF Research Database (Denmark)

    Jensen, Anders; Fagö-Olsen, Helena; Sørensen, Christian Hjort

    2013-01-01

    and included species that are considered putative pathogens in periodontal diseases, i.e. Porphyromonas gingivalis, Porphyromonas endodontalis, and Tannerella forsythia. Unifrac analysis showed that recurrent tonsillitis is associated with a shift in the microbiota of the tonsillar crypts. Fusobacterium...... were almost exclusively detected in children. In contrast, Streptococcus pseudopneumoniae was present in all samples. Obligate anaerobes like Porphyromonas, Prevotella, and Fusobacterium were abundantly present in children, but the species diversity of Porphyromonas and Prevotella was larger in adults...

  9. Association of oncogenic bacteria with colorectal cancer in South China.

    Science.gov (United States)

    Zhou, Youlian; He, Hanchang; Xu, Haoming; Li, Yingfei; Li, Zhiming; Du, Yanlei; He, Jie; Zhou, Yongjian; Wang, Hong; Nie, Yuqiang

    2016-12-06

    To quantify Fusobacterium spp., Enterococcus faecalis (E.faecalis), Enterotoxigenic Bacteroides fragilis (ETBF), and Enteropathogenic Escherichia coli in colorectal cancer (CRC) patients and their possible association with CRC clinicopathogical features, we collected the resected tumors and adjacent normal tissues (N) from 97 CRC patients. 48 age- and sex-matched healthy controls (HC) were also recruited. Real-time PCR was used for bacterial quantification. The median abundance ofFusobacterium spp.(p colon tissue in the proximity of the tumor.

  10. Tetracycline resistance and TetM in oral anaerobic bacteria and Neisseria perflava-N. sicca.

    OpenAIRE

    Roberts, M C; Moncla, B J

    1988-01-01

    Tetracycline-resistant organisms isolated from six patients with periodontal disease included Bacteroides spp., Eubacterium spp., Fusobacterium nucleatum, Neisseria perflava-N. sicca, Peptostreptococcus anaerobius, Veillonella parvula, and facultative streptococci. All but the Bacteroides spp. and Eubacterium spp. hybridized with the TetM determinant. An additional 417 bacterial strains were screened, and 4% of both the oral streptococci and the Fusobacterium spp. hybridized with the TetM probe.

  11. Bacterially-Associated Transcriptional Remodelling in a Distinct Genomic Subtype of Colorectal Cancer Provides a Plausible Molecular Basis for Disease Development.

    Directory of Open Access Journals (Sweden)

    Katie S Lennard

    Full Text Available The relevance of specific microbial colonisation to colorectal cancer (CRC disease pathogenesis is increasingly recognised, but our understanding of possible underlying molecular mechanisms that may link colonisation to disease in vivo remains limited. Here, we investigate the relationships between the most commonly studied CRC-associated bacteria (Enterotoxigenic Bacteroides fragilis, pks+ Escherichia coli, Fusobacterium spp., afaC+ E. coli, Enterococcus faecalis & Enteropathogenic E. coli and altered transcriptomic and methylation profiles of CRC patients, in order to gain insight into the potential contribution of these bacteria in the aetiopathogenesis of CRC. We show that colonisation by E. faecalis and high levels of Fusobacterium is associated with a specific transcriptomic subtype of CRC that is characterised by CpG island methylation, microsatellite instability and a significant increase in inflammatory and DNA damage pathways. Analysis of the significant, bacterially-associated changes in host gene expression, both at the level of individual genes as well as pathways, revealed a transcriptional remodeling that provides a plausible mechanistic link between specific bacterial colonisation and colorectal cancer disease development and progression in this subtype; these included upregulation of REG3A, REG1A and REG1P in the case of high-level colonization by Fusobacterium, and CXCL10 and BMI1 in the case of colonisation by E. faecalis. The enrichment of both E. faecalis and Fusobacterium in this CRC subtype suggests that polymicrobial colonisation of the colonic epithelium may well be an important aspect of colonic tumourigenesis.

  12. Estimation of bacterial hydrogen sulfide production in vitro

    Directory of Open Access Journals (Sweden)

    Amina Basic

    2015-06-01

    Full Text Available Oral bacterial hydrogen sulfide (H2S production was estimated comparing two different colorimetric methods in microtiter plate format. High H2S production was seen for Fusobacterium spp., Treponema denticola, and Prevotella tannerae, associated with periodontal disease. The production differed between the methods indicating that H2S production may follow different pathways.

  13. Influence of age and immunization on development of gingivitis in rats

    DEFF Research Database (Denmark)

    Lekic, P; Klausen, B; Friis-Hasché, E

    1989-01-01

    To study the effect of age and antigenic priming on the development of gingivitis, 33 healthy rats were placed in contact with Streptococcus mutans, Actinomyces viscosus, Fusobacterium nucleatum, and Bacteroides gingivalis. On days 0, 3, 7, and 14 after inoculation, the gingival condition...

  14. Microbiology and management of joint and bone infections due to anaerobic bacteria.

    Science.gov (United States)

    Brook, Itzhak

    2008-03-01

    To describes the microbiology, diagnosis, and management of septic arthritis and osteomyelitis due to anaerobic bacteria. The predominant anaerobes in arthritis are anaerobic Gram-negative bacilli (AGNB) including the Bacteroides fragilis group, Fusobacterium spp., Peptostreptococcus spp., and Propionibacterium acnes. Infection with P. acnes is associated with a prosthetic joint, previous surgery, and trauma. B. fragilis group is associated with distant infection, Clostridium spp. with trauma, and Fusobacterium spp. with oropharyngeal infection. Most cases of anaerobic arthritis, in contrast to anaerobic osteomyelitis, involved a single isolate, and most cases are secondary to hematogenous spread. The predominant anaerobes in osteomyelitis are Bacteroides, Peptostreptococcus, Fusobacterium, and Clostridium spp. as well as P. acnes. Conditions predisposing to bone infections are vascular disease, bites, contiguous infection, peripheral neuropathy, hematogenous spread, and trauma. Pigmented Prevotella and Porphyromonas spp. are mostly isolated in skull and bite infections, members of the B. fragilis group in hand and feet infections, and Fusobacterium spp. in skull, bite, and hematogenous long bone infections. Many patients with osteomyelitis due to anaerobic bacteria have evidence of an anaerobic infection elsewhere in the body that is the source of the organisms involved in the osteomyelitis. Treatment of arthritis and osteomyelitis involving anaerobic bacteria includes symptomatic therapy, immobilization in some cases, adequate drainage of purulent material, and antibiotic therapy effective against these organisms. Anaerobic bacteria can cause septic arthritis and osteomyelitis. Correct diagnosis and appropriate therapy are important contributor to successful outcome.

  15. Development of Vaccines to Prevent Wound Infections due to Anaerobic Bacteria

    Science.gov (United States)

    1980-08-01

    containing an inoculum that included anaerobes ( Eubacterium , Clostridium, Bacteroides, Peptococcus, and Fusobacterium) and aerobes (E. coli, enterococcus) in a...serogroups of the Bacteroides spp . based on 0-antigens have been described. (11-13) Type-specific antisera raised in rabbits against twenty distinct

  16. Applying Nano technology to Human Health: Revolution in Biomedical Sciences

    International Nuclear Information System (INIS)

    Shrivastava, S.; Dash, D.

    2009-01-01

    Recent research on bio systems at the nano scale has created one of the most dynamic science and technology domains at the confluence of physical sciences, molecular engineering, biology, biotechnology, and medicine. This domain includes better understanding of living and thinking systems, revolutionary biotechnology processes, synthesis of new drugs and their targeted delivery, regenerative medicine, necrophorum engineering, and developing a sustainable environment. Nano bio systems research is a priority in many countries and its relevance within nano technology is expected to increase in the future. The realisation that the nano scale has certain properties needed to solve important medical challenges and cater to unmet medical needs is driving nano medical research. The present review explores the significance of nano science and latest nano technologies for human health. Addressing the associated opportunities, the review also suggests how to manage far-reaching developments in these areas

  17. BACTERIAL PROFILES FOR CHRONIC AND AGGRESSIVE PERIODONTITIS IN A SAMPLE POPULATION GROUP. A CROSS-SECTIONAL STUDY

    Directory of Open Access Journals (Sweden)

    Alexandra-Cornelia TEODORESCU

    2017-06-01

    Full Text Available Aim. The study aims at determining some possible significant differences in the subgingival microbial profiles of patients with generalized chronic periodontitis (GCP and generalized aggressive periodontitis (GAP, as a tool in helping with differential diagnostic. Materials and methods. 20 subgingival fluid samples (10 from GAP and 10 from GCP patients were subjected to a Real-Time Polymerase Chain Reaction technique in order to determine the prevalence and the counts of 9 periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tanerella forsythia, Prevotella intermedia, Peptostreptococcus micros, Fusobacterium nucleatum, Eubacterium nodatum and Capnocytophaga gingivalis. Results and discussion. Fusobacterium nucleatum was singnificantly correlated with the aggressive periodontitis group, but no significant differences were found for the other 8 periodontal bacteria. Conclusions. The prevalence or count of some periodontal pathogens could help clinicians make an easier differential diagnostic between GCP and GAP, however further studies, conducted on larger population samples, are still needed.

  18. Acute Necrotizing Ulcerative Gingivitis: Microbial and Immunologic Studies

    Science.gov (United States)

    1983-10-01

    Fusobacterium nucleatum and Bacteroides strains isolated from ANUG patients, chronic periodontitis patients, non-oral abscesses and healthy gingival sulci...34 Odd ii 4n....00p IdaOdifir 0’ block flumber) Periodontal Disease Spirochetes Microbiology Fusobacteria immunology Bacteroides 20. AN3T7RAC~r C4.tft...and serological etudies with ’We 7. nucleatum isolates from patients with ANUG, chronic periodontitis , Juvenile periodontitis , and adults and children

  19. Regulation of Ghrelin Receptor by Periodontal Bacteria In Vitro and In Vivo

    OpenAIRE

    Nokhbehsaim, Marjan; Damanaki, Anna; Nogueira, Andressa Vilas Boas; Eick, Sigrun; Memmert, Svenja; Zhou, Xiaoyan; Nanayakkara, Shanika; Götz, Werner; Cirelli, Joni Augusto; Jäger, Andreas; Deschner, James

    2017-01-01

    Ghrelin plays a major role in obesity-related diseases which have been shown to be associated with periodontitis. This study sought to analyze the expression of the functional receptor for ghrelin (GHS-R1a) in periodontal cells and tissues under microbial conditions in vitro and in vivo. The GHS-R1a expression in human periodontal cells challenged with the periodontopathogen Fusobacterium nucleatum, in gingival biopsies from periodontally healthy and diseased individuals, and from rats with a...

  20. Isolation of obligate and facultative anaerobic bacteria from feline subcutaneous abscesses.

    Science.gov (United States)

    Hoshuyama, S; Kanoe, M; Amimoto, A

    1996-03-01

    A total of 113 specimens collected from purulent skin lesions of household cats was examined bacteriologically. Ninety seven isolates obtained from 74 specimens (65.5%). Of these, 11 specimens (9.7%) contained obligate anaerobes only, 18 specimens (15.9%) yielded both obligate and facultative anaerobes. In the obligate anaerobes detected, genus Fusobacterium was the most frequently observed and F. nucleatum was most common species. Pasteurella multocida was the facultative anaerobe which was most frequently detected.

  1. Detection of a Mixed Infection in a Culture-Negative Brain Abscess by Broad-Spectrum Bacterial 16S rRNA Gene PCR ▿ †

    Science.gov (United States)

    Keller, Peter M.; Rampini, Silvana K.; Bloemberg, Guido V.

    2010-01-01

    We describe the identification of two bacterial pathogens from a culture-negative brain abscess by the use of broad-spectrum 16S rRNA gene PCR. Simultaneous detection of Fusobacterium nucleatum and Porphyromonas endodontalis was possible due to a 24-bp length difference of their partially amplified 16S rRNA genes, which allowed separation by high-resolution polyacrylamide gel electrophoresis. PMID:20392909

  2. Increased activity of a new chlorofluoroquinolone, BAY y 3118, compared with activities of ciprofloxacin, sparfloxacin, and other antimicrobial agents against anaerobic bacteria.

    OpenAIRE

    Aldridge, K E

    1994-01-01

    A total of 435 clinical isolates of anaerobes were tested with a broth microdilution method to determine the activity of BAY y 3118 compared with those of other agents against anaerobic bacteria. All strains of Bacteroides capillosus, Prevotella spp., Porphyromonas spp., Fusobacterium spp., Clostridium spp., Eubacterium spp., Peptostreptococcus spp., and Veillonella parvula were susceptible (MICs of < or = 2 micrograms/ml) to BAY y 3118. Against the 315 strains of the Bacteroides fragilis gro...

  3. In vitro activity of cefbuperazone, a new cephamycin, against anaerobic bacteria.

    OpenAIRE

    Prabhala, R H; Thadepalli, H; Rao, B; Bansal, M B; Marshall, R

    1985-01-01

    The 90% MIC of cefbuperazone (BMY 25182) was 32 micrograms/ml for Bacteroides fragilis and Bacteroides spp., 128 micrograms/ml for Fusobacterium and Clostridium spp., 64 micrograms/ml for Eubacterium and Peptococcus spp., 8 micrograms/ml for Actinomyces spp., and 32 micrograms/ml for Peptostreptococcus spp. The level of activity of cefbuperazone was higher against B. fragilis and lower against anaerobic cocci than those of related cephalosporins, i.e., cefoxitin, cefoperazone, cefotaxime, cef...

  4. An unclassified Eubacterium taxon in acute dento-alveolar abscess.

    Science.gov (United States)

    Wade, W G; Lewis, M A; Cheeseman, S L; Absi, E G; Bishop, P A

    1994-02-01

    The microflora of pus samples aspirated from 50 acute dento-alveolar abscesses was examined. A total of 143 bacterial strains was isolated, consisting predominantly of Prevotella spp., alpha-haemolytic Streptococcus spp., Peptostreptococcus spp. and Eubacterium spp. An unclassified asaccharolytic Eubacterium taxon was encountered in 17 (34%) of the abscesses. This taxon was found to have a positive association with Fusobacterium spp. and a negative association with alpha-haemolytic Streptococcus spp.

  5. Quantitative Analysis of Intestinal Flora of Uygur and Han Ethnic Chinese Patients with Ulcerative Colitis

    Directory of Open Access Journals (Sweden)

    Ping Yao

    2016-01-01

    Full Text Available Aim. To study the correlation between intestinal flora and ulcerative colitis by analyzing the abundance of Bacteroides, Fusobacterium, Clostridium, Bifidobacterium spp., and Faecalibacterium prausnitzii in the intestinal of ulcerative colitis (UC patients and healthy controls with Uygur and Han ethnic. Methods. Bacterial genomic DNA was extracted from fecal samples and analyzed with real-time fluorescence quantitative polymerase chain reaction (PCR to identify the abundance of Bacteroides, Fusobacterium, Clostridium, Bifidobacterium spp., and Faecalibacterium prausnitzii. Results. The samples from UC patients, Uygur and Han ethnic combined, had higher abundance of Bacteroides (P=0.026 but lower Clostridium (P=0.004, Bifidobacterium spp. (P=0.009, and Faecalibacterium prausnitzii (P=0.008 than those from healthy controls. Among UC patients, Bacteroides population was raised in acute UC patients (P≤0.05, while the abundance of Clostridium, Bifidobacterium spp., Fusobacterium, and Faecalibacterium prausnitzii decreased (P≤0.05 compared with the remission. In both UC patients group and control group, no difference was observed in the abundance of these 5 bacteria between the Han and the Uygur group. Conclusions. Variations in the abundance of these five bacterial strains in intestines may be associated with the occurrence of UC in Uygur and Han populations; however, these variations were not associated with ethnic difference.

  6. Anaerobic bacteria in 118 patients with deep-space head and neck infections from the University Hospital of Maxillofacial Surgery, Sofia, Bulgaria.

    Science.gov (United States)

    Boyanova, Lyudmila; Kolarov, Rossen; Gergova, Galina; Deliverska, Elitsa; Madjarov, Jivko; Marinov, Milen; Mitov, Ivan

    2006-09-01

    The aim of this study was to assess the incidence and susceptibility to antibacterial agents of anaerobic strains in 118 patients with head and neck abscesses (31) and cellulitis (87). Odontogenic infection was the most common identified source, occurring in 73 (77.7%) of 94 patients. The incidence of anaerobes in abscesses and cellulitis was 71 and 75.9%, respectively, and that in patients before (31 patients) and after (87) the start of empirical treatment was 80.6 and 72.4%, respectively. The detection rates of anaerobes in patients with odontogenic and other sources of infection were 82.2 and 71.4%, respectively. In total, 174 anaerobic strains were found. The predominant bacteria were Prevotella (49 strains), Fusobacterium species (22), Actinomyces spp. (21), anaerobic cocci (20) and Eubacterium spp. (18). Bacteroides fragilis strains were isolated from 7 (5.9%) specimens. The detection rate of Fusobacterium strains from non-treated patients (32.2%) was higher than that from treated patients (13.8%). Resistance rates to clindamycin and metronidazole of Gram-negative anaerobes were 5.4 and 2.5%, respectively, and those of Gram-positive species were 4.5 and 58.3%, respectively. One Prevotella strain was intermediately susceptible to ampicillin/sulbactam. In conclusion, the start of empirical treatment could influence the frequency or rate of isolation of Fusobacterium species. The involvement of the Bacteroides fragilis group in some head and neck infections should be considered.

  7. Safety, Clinical Response, and Microbiome Findings Following Fecal Microbiota Transplant in Children With Inflammatory Bowel Disease.

    Science.gov (United States)

    Goyal, Alka; Yeh, Andrew; Bush, Brian R; Firek, Brian A; Siebold, Leah M; Rogers, Matthew Brian; Kufen, Adam D; Morowitz, Michael J

    2018-01-18

    The role of fecal microbiota transplant (FMT) in the treatment of pediatric inflammatory bowel disease (IBD) is unknown. The aims of this study were to assess safety, clinical response, and gut microbiome alterations in children with Crohn's disease (CD), ulcerative colitis (UC), or indeterminate colitis (IC). In this open-label, single-center prospective trial, patients with IBD refractory to medical therapy underwent a single FMT by upper and lower endoscopy. Adverse events, clinical response, gut microbiome, and biomarkers were assessed at baseline, 1 week, 1 month, and 6 months following FMT. Twenty-one subjects were analyzed, with a median age of 12 years, of whom 57% and 28% demonstrated clinical response at 1 and 6 months post-FMT, respectively. Two CD patients were in remission at 6 months. Adverse events attributable to FMT were mild to moderate and self-limited. Patients prior to FMT showed decreased species diversity and significant microbiome compositional differences characterized by increased Enterobacteriaceae, Enterococcus, Haemophilus, and Fusobacterium compared with donors and demonstrated increased species diversity at 30 days post-FMT. At 6 months, these changes shifted toward baseline. Clinical responders had a higher relative abundance of Fusobacterium and a lower diversity at baseline, as well as a greater shift toward donor-like microbiome after FMT compared with nonresponders. A single FMT is relatively safe and can result in a short-term response in young patients with active IBD. Responders possessed increased Fusobacterium prior to FMT and demonstrated more significant microbiome changes compared with nonresponders after FMT. Microbiome characteristics may help in predicting response. © 2018 Crohn’s & Colitis Foundation of America. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

  8. Estudo bacteriológico de lesões periapicais

    Directory of Open Access Journals (Sweden)

    ROCHA Márcia Maria de Negreiros Pinto

    1998-01-01

    Full Text Available O tecido perirradicular de 30 casos cirúrgicos foi submetido a exame microbiológico e histopatológico. Foram isoladas 137 cepas bacterianas das 30 lesões periapicais estudadas. Do total de bactérias isoladas, 90 (65,7% foram caracterizadas como anaeróbias estritas, 40 (29,2% como anaeróbias facultativas e 7 (5,1% como aeróbias estritas. Fusobacterium nucleatum, bacilos Gram-negativos pigmentados anaeróbios estritos, Peptostreptococcus sp, Streptococcus mitis e bacilos Gram-positivos não esporulados anaeróbios estritos foram, em ordem decrescente de freqüência, as bactérias mais comumente isoladas das lesões periapicais. A análise histopatológica demonstrou prevalência de granuloma periapical. O estudo de sensibilidade a antimicrobianos foi realizado em 16 cepas de Fusobacterium nucleatum. Os antimicrobianos utilizados foram penicilina, cefoxitina, eritromicina, metronidazol e tetraciclina. Todas as amostras apresentaram suscetibilidade à cefoxitina, tetraciclina e metronidazol; os percentuais de resistência à penicilina e à eritromicina foram 12,5 e 68,8, respectivamente. Os percentuais de resistência de Fusobacterium nucleatum, uma espécie bacteriana importante na patogênese das lesões periapicais, à penicilina e à eritromicina, enfatizam a necessidade de outros estudos de resistência a antimicrobianos de bactérias isoladas em infecções odontogênicas.

  9. Influence of complement on neutrophil extracellular trap release induced by bacteria

    DEFF Research Database (Denmark)

    Palmer, Lisa Joanne; Damgaard, Christian; Holmstrup, Palle

    2016-01-01

    by Staphylococcus aureus and three oral bacteria: Actinomyces viscosus, Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum subsp. vincettii. Material and Methods Bacteria-stimulated NET release from the neutrophils of healthy donors was measured fluorometrically. Various complement containing...... In conclusion, complement opsonization promotes NET release induced by a variety of bacteria, including A. actinomycetemcomitans, and CR1 plays a dominant role in the process. Complement consumption or deficiency may compromise NETosis induced by some bacterial species, including A. actinomycetemcomitans....... Within biofilms, the complement-inactivating abilities of some bacteria may protect other species against NETosis, while these are more vulnerable when adopting a planktonic lifestyle....

  10. Actividad “in vitro” de 10 antimicrobianos frente a bacterias anaerobias: Estudio multicéntrico, 1999-2002 “In vitro” activity of ten antimicrobial agents against anaerobic bacteria. A collaborative study, 1999-2002

    Directory of Open Access Journals (Sweden)

    M. Litterio

    2004-09-01

    Full Text Available Se evaluó la actividad de ampicilina, ampicilina-sulbactama, cefoxitina, ceftriaxona, imipenem, piperacilina, piperacilina-tazobactama, clindamicina, metronidazol y azitromicina frente a 166 cepas de bacterias anaerobias aisladas en 8 hospitales de Buenos Aires. Se estudiaron: Bacteroides grupo fragilis (65, Fusobacterium spp. (26, Prevotella spp. (21, Porphyromonas spp. (10, Clostridium difficile (10, otros clostridios (12 y cocos gram-positivos (22. Las CIMs se determinaron usando el método patrón de dilución en agar recomendado por el NCCLS, documento M11-A5. Los antibióticos más activos fueron metronidazol y piperacilina-tazobactama que exhibieron valores de CIM90£ 2 µg/ml y £ 4 µg/ml frente a los microorganismos gram-negativos y £ 2 µg/ml y £ 8 µg/ml frente a los microorganismos gram-positivos, respectivamente. Entre los b-lactámicos el orden de actividad frente a bacilos gram-negativos fue: imipenem > piperacilina > cefoxitina > ceftriaxona > ampicilina. En gram-positivos la actividad decreciente fue: piperacilina> imipenem > cefoxitina > ceftriaxona > ampicilina. La mayoría de las especies estudiadas mostraron distintos niveles de resistencia con clindamicina y azitromicina. Sin embargo, el 90% de las cepas de Fusobacterium nucleatum y Por-phyromonas spp. fue inhibido por una concentración de 0,125 µg/ml de clindamicina y azitromicina, respectivamente.The antimicrobial activity of ampicillin, ampicillin-sulbactam, cefoxitin, ceftriaxone, imipenem, piperacillin, piperacillin-tazobactam, clindamycin, metronidazole, and azitromycin was assesed against 166 strains of anaerobic bacteria recovered from eight hospitals in Buenos Aires. The strains studied were Bacteroidesfragilis group (65, Fusobacterium spp. (26, Prevotella spp. (21, Porphyromonas spp. (10, Clostridium difficile (10, other clostridia (12, and gram-positive cocci (22. The MICs were determined by the agar dilution method according to NCCLS document M11-A5

  11. Association between periodontal condition and subgingival microbiota in women during pregnancy: a longitudinal study.

    Science.gov (United States)

    Borgo, Priscila Viola; Rodrigues, Viviane Aparecida Arenas; Feitosa, Alfredo Carlos Rodrigues; Xavier, Karla Correa Barcelos; Avila-Campos, Mario Julio

    2014-01-01

    In this study, the gingival conditions and the quantitative detection for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in pregnant women were determined. Quantitative determinations of periodontal bacteria by using a SyBr green system in women during pregnancy were performed. Women at the 2nd and 3rd trimesters of pregnancy and non-pregnant women were included in this study. A. actinomycetemcomitans was observed in high numbers in women at the 2nd and 3rd trimesters of pregnancy with a significant difference (pperiodontal disease.

  12. Metagenomic analysis of faecal microbiome as a tool towards targeted non-invasive biomarkers for colorectal cancer

    DEFF Research Database (Denmark)

    Yu, Jun; Feng, Qiang; Wong, Sunny Hei

    2017-01-01

    known associations of Fusobacterium nucleatum and Peptostreptococcus stomatis with CRC, we found significant associations with several species, including Parvimonas micra and Solobacterium moorei. We identified 20 microbial gene markers that differentiated CRC and control microbiomes, and validated 4...... in the independent Chinese cohort with AUC=0.84 and OR of 23. These genes were enriched in early-stage (I-II) patient microbiomes, highlighting the potential for using faecal metagenomic biomarkers for early diagnosis of CRC. CONCLUSIONS: We present the first metagenomic profiling study of CRC faecal microbiomes...

  13. Fermentation of mucins and plant polysaccharides by anaerobic bacteria from the human colon.

    Science.gov (United States)

    Salyers, A A; West, S E; Vercellotti, J R; Wilkins, T D

    1977-01-01

    A total of 154 strains from 22 species of Bifidobacterium, Peptostreptococcus, Lactobacillus, Ruminococcus, Coprococcus, Eubacterium, and Fusobacterium, which are present in high concentrations in the human colon, were surveyed for their ability to ferment 21 different complex carbohydrates. Plant polysaccharides, including amylose, amylopectin, pectin, polygalacturonate, xylan, laminarin, guar gum, locust bean gum, gum ghatti, gum arabic, and gum tragacanth, were fermented by some strains from Bifidobacterium, Peptostreptococcus, Ruminococcus, and Eubacterium species. Porcine gastric mucin, which was fermented by some strains of Ruminococcus torques and Bifidobacterium bifidum, was the only mucin utilized by any of the strains tested. PMID:563214

  14. Occurrence of periodontal bacteria in atheroma coronary arteries from patients with chronic periodontitis

    OpenAIRE

    Silvia Linard Marcelino

    2008-01-01

    Hoje em dia existe um consenso de que a prevenção e o tratamento de doenças periodontais são importantes na redução de mortalidade e morbidade associadas à doença cardíaca, e uma correlação entre aterosclerose e doença periodontal ainda é desconhecida. Neste estudo, a presença de Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, Prevotella nigrescens, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Campyloba...

  15. Brain abscess in a Japanese Black calf: Utility of computed tomography (CT)

    International Nuclear Information System (INIS)

    El-Khodery, S.; Yamada, K.; Aoki, D.; Kamio, K.; Kishimoto, M.; Shimizu, J.; Kobayashi, Y.; Ishii, M.; Inokuma, H.; Yamauchi, S.; Matsui, T.

    2008-01-01

    Computed tomography (CT) was used for diagnosis of brain abscess in a 6-month-old, Japanese Black calf presented with neurological dysfunction, compulsive circling and vision disturbance. CT images showed asymmetric lateral ventricles, and presence of intra-cranial multiple low absorption lesions surrounded by capsule suggestive of abscess in the right cerebral hemisphere. Postmortem examination revealed marked swelling of right cerebral hemisphere and olfactory bulb. Multilocular large abscess containing creamy pus was found to occupy most area of periventricular and lateral ventricle. Fusobacterium necrophrum was isolated from the abscess contents as the causative agent. These results demonstrate that CT is useful tool for tentative diagnosis of bovine brain abscess

  16. An evaluation of the seasonality of veterinary treatments for lameness in UK dairy cattle.

    Science.gov (United States)

    Laven, R A; Lawrence, K R

    2006-10-01

    Of the 4 main diseases causing lameness in UK dairy cattle, 2 (digital dermatitis and foul-in-the-foot, or interdigital necrobacillosis) are infectious diseases that are commonly associated with similar underfoot conditions, and 2 (sole ulcer and white-line disease) result from subclinical laminitis. Comparison of the seasonality of these 2 diseases can identify whether their own unique risk factors are more important than the risk factors they share. Using a database from 46 veterinarians distributed across the United Kingdom, the seasonality of treatments for these 4 diseases was evaluated. This analysis showed that the seasonality of lameness in UK dairy cattle was significantly reduced in comparison with previous reports from the 1980s. This was primarily due to 1) a reduction in the seasonality of digital dermatitis (outbreaks of which accounted for 6.7% of veterinary reports), with only 60% of reports during the winter as opposed to 72% in earlier studies, and 2) a change in the seasonality of white-line disease from a disease that was most commonly seen in the winter (55% of reports) to a disease that was most commonly reported in the late summer or early autumn, with significantly more reports in August and October than in February. Comparison of the seasonality of digital dermatitis and foul-in-the-foot showed a significant difference in the seasonality of the 2 diseases, with foul-in-the-foot being significantly more common compared with digital dermatitis in June and August. This implies that significant risk factors exist for digital dermatitis that are not as important for foul-in-the-foot. The difference in the seasonality of sole ulcer and white-line disease was even more marked, with white-line disease being significantly more common than sole ulcer from August to October, despite these diseases being more closely linked in the literature than digital dermatitis and foul-in-the-foot. This comparison shows that the seasonality of lameness has changed

  17. [Study on Microbial Diversity of Peri-implantitis Subgingival by High-throughput Sequencing].

    Science.gov (United States)

    Li, Zhi-jie; Wang, Shao-guo; Li, Yue-hong; Tu, Dong-xiang; Liu, Shi-yun; Nie, Hong-bing; Li, Zhi-qiang; Zhang, Ju-mei

    2015-07-01

    To study microbial diversity of peri-implantitis subgingival with high-throughput sequencing, and investigate microbiological etiology of peri-implantitis. Subgingival plaques were sampled from the patients with peri-implantitis (D group) and non-peri-implantitis subjects (N group). The microbiological diversity of the subgingival plaques was detected by sequencing V4 region of 16S rRNA with Illumina Miseq platform. The diversity of the community structure was analyzed using Mothur software. A total of 156 507 gene sequences were detected in nine samples and 4 402 operational taxonomic units (OTUs) were found. Selenomonas, Pseudomonas, and Fusobacterium were dominant bacteria in D group, while Fusobacterium, Veillonella and Streptococcus were dominant bacteria in N group. Differences between peri-implantitis and non-peri-implantitis bacterial communities were observed at all phylogenetic levels by LEfSe, which was also found in PcoA test. The occurrence of peri-implantitis is not only related to periodontitis pathogenic microbe, but also related with the changes of oral microbial community structure. Treponema, Herbaspirillum, Butyricimonas and Phaeobacte may be closely related to the occurrence and development of peri-implantitis.

  18. In vitro activity of FK037, a new parenteral cephalosporin, against anaerobic bacteria.

    Science.gov (United States)

    Kato, N; Kato, H; Tanaka, Y; Bando, K; Watanabe, K; Ueno, K

    1993-01-01

    The activity of FK037, a new parenteral cephalosporin, was compared with those of cefpirome, ceftazidime, and flomoxef against 322 recent clinical isolates of anaerobic bacteria. A fastidious facultative anaerobe, Gardnerella vaginalis, was also studied. FK037 inhibited 90% of isolates of Peptostreptococcus anaerobius, Peptostreptococcus asaccharolyticus, Clostridium perfringens, Mobiluncus spp., G. vaginalis, and Porphyromonas gingivalis at 200 micrograms/ml, respectively; the activity of FK037 was comparable to those of cefpirome and ceftazidime but lower than that of flomoxef. The activity of FK037 against Fusobacterium nucleatum, Fusobacterium varium, and Bilophila wadsworthia decreased when inoculum size was increased from 10(6) to 10(8) CFU/ml. Little influence of inoculum size on the activity of FK037 was observed for other isolates tested. Medium pH affected the activity of FK037 against F. varium (MICs at pHs 5 and 7, 3.13 and 100 micrograms/ml, respectively) and Bacteroides gracilis (MICs at pHs 5 and 7, 12.5 and 1.56 micrograms/ml, respectively) but not against other organisms tested. FK037 was less resistant than flomoxef to hydrolysis by beta-lactamase group 2e derived from B. fragilis GAI 0558 and GAI 10150. PMID:8517721

  19. The relationship between turbidity of mouth-rinsed water and oral health status.

    Science.gov (United States)

    Takeuchi, Susumu; Ueno, Masayuki; Takehara, Sachiko; Pham, Thuy Anh Vu; Hakuta, Chiyoko; Morishima, Seiji; Shinada, Kayoko; Kawaguchi, Yoko

    2013-01-01

    The purpose of this study was to examine the relationship between turbidity of mouth rinsed water and oral health status such as dental and periodontal conditions, oral hygiene status, flow rate of saliva and oral bacteria. Subjects were 165 patients who visited the Dental Hospital, Tokyo Medical and Dental University. Oral health status, including dental and periodontal conditions, oral hygiene status and flow rate of saliva, was clinically examined. The turbidity was measured with a turbidimeter. Quantification of Fusobacterium spp, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and total bacteria levels was performed using real-time PCR. The Pearson correlation and multiple regression analysis were used to explore the associations between the turbidity and oral health parameters. The turbidity showed significant correlations with the number of decayed teeth and deep pockets, the plaque index, extent of tongue coating and Fusobacterium spp, P. gingivalis, T. forsythia, T. denticola and total bacteria levels. In a multiple regression model, the turbidity was negatively associated with the flow rate of saliva and positively associated with the total number of bacteria (p turbidity of mouth rinsed water could be used as an indicator to evaluate oral health condition and the amount of bacteria in the oral cavity. In addition, the turbiditimeter appeared as a simple and objective device for screening abnormality of oral health condition at chair side as well as community-based research.

  20. Pyrosequencing Analysis of Subgingival Microbiota in Distinct Periodontal Conditions.

    Science.gov (United States)

    Park, O-J; Yi, H; Jeon, J H; Kang, S-S; Koo, K-T; Kum, K-Y; Chun, J; Yun, C-H; Han, S H

    2015-07-01

    Subgingival microorganisms are potentially associated with periodontal diseases. However, changes in the subgingival microbiota during the progress of periodontal diseases are poorly understood. In this study, we analyzed bacterial communities in the subgingival paper point samples from 32 Korean individuals with no sign of disease, gingivitis, or periodontitis using 454 FLX Titanium pyrosequencing. A total of 256,113 reads representing 26 phyla, 433 genera, and 1,016 species were detected. Bacteroidetes, Fusobacteria, Synergistetes, and Spirochaetes were the abundant phyla in periodontitis subjects, whereas Firmicutes and Proteobacteria were identified as the dominant phyla in the gingivitis and healthy subjects, respectively. Although high levels of Porphyromonas, Fusobacterium, Fretibacterium, Rothia, Filifactor, and Treponema genera were observed in the periodontitis subjects, Streptococcus, Capnocytophaga, Leptotrichia, and Haemophilus genera were found at high frequency in the gingivitis subjects. Species including Porphyromonas gingivalis, Fusobacterium nucleatum, and Fretibacterium fastidiosum were significantly increased in periodontitis subjects. On the other hand, Streptococcus pseudopneumoniae, Haemophilus parainfluenzae, and Leptotrichia hongkongensis were preferentially observed in the gingivitis subjects. Intriguingly, the halophile Halomonas hamiltonii was revealed as a predominant species in the healthy subjects. Based on Fast UniFrac analysis, distinctive bacterial clusters were classified for the healthy, gingivitis, and periodontitis state. The current findings might be useful for understanding the pathogenesis, diagnosis, and treatment of periodontal diseases. © International & American Associations for Dental Research 2015.

  1. Antibiotic resistance genes in anaerobic bacteria isolated from primary dental root canal infections.

    Science.gov (United States)

    Rôças, Isabela N; Siqueira, José F

    2012-12-01

    Fourty-one bacterial strains isolated from infected dental root canals and identified by 16S rRNA gene sequence were screened for the presence of 14 genes encoding resistance to beta-lactams, tetracycline and macrolides. Thirteen isolates (32%) were positive for at least one of the target antibiotic resistance genes. These strains carrying at least one antibiotic resistance gene belonged to 11 of the 26 (42%) infected root canals sampled. Two of these positive cases had two strains carrying resistance genes. Six out of 7 Fusobacterium strains harbored at least one of the target resistance genes. One Dialister invisus strain was positive for 3 resistance genes, and 4 other strains carried two of the target genes. Of the 6 antibiotic resistance genes detected in root canal strains, the most prevalent were blaTEM (17% of the strains), tetW (10%), and ermC (10%). Some as-yet-uncharacterized Fusobacterium and Prevotella isolates were positive for blaTEM, cfxA and tetM. Findings demonstrated that an unexpectedly large proportion of dental root canal isolates, including as-yet-uncharacterized strains previously regarded as uncultivated phylotypes, can carry antibiotic resistance genes. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. Antimicrobial resistance determinants among anaerobic bacteria isolated from footrot.

    Science.gov (United States)

    Lorenzo, María; García, Nuria; Ayala, Juan Alfonso; Vadillo, Santiago; Píriz, Segundo; Quesada, Alberto

    2012-05-25

    Antibiotic resistance has been evaluated among 36 Gram negative and anaerobic bacilli (10 Bacteroides, 11 Prevotella, 7 Porphyromonas and 8 Fusobacterium strains) isolated from clinical cases of caprine and ovine footrot (necrotic pododermatitis). The initial analysis on this bacterial consortium evaluates the relationships existing among antimicrobial resistance determinants, phenotype expression and mobilization potential. The Bacteroides strains were generally resistant to penicillins, first-generation cephalosporins, tetracycline and erythromycin, and expressed low level of β-lactamase activity. The main determinants found among the Bacteroides strains were cepA and tetQ genes, conferring resistance to β-lactams and tetracycline, respectively. A general susceptibility to β-lactams was shown for most Prevotella, Porphyromonas and Fusobacterium strains, where none of the β-lactamase genes described in Bacteroides was detected. Resistance to tetracycline and/or erythromycin was found among the three bacterial groups. Although tetQ genes were detected for several Prevotella and Porphyromonas strains, a unique ermF positive was revealed among Prevotella strains. The expression of resistance markers was not related with the polymorphism of their coding sequences. However, the finding of sequence signatures for conjugative transposons in the vicinities of tetQ and ermF suggests a mobilization potential that might have contributed to the spread of antimicrobial resistance genes. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. Oral Gram-negative anaerobic bacilli as a reservoir of β-lactam resistance genes facilitating infections with multiresistant bacteria.

    Science.gov (United States)

    Dupin, Clarisse; Tamanai-Shacoori, Zohreh; Ehrmann, Elodie; Dupont, Anais; Barloy-Hubler, Frédérique; Bousarghin, Latifa; Bonnaure-Mallet, Martine; Jolivet-Gougeon, Anne

    2015-02-01

    Many β-lactamases have been described in various Gram-negative bacilli (Capnocytophaga, Prevotella, Fusobacterium, etc.) of the oral cavity, belonging to class A of the Ambler classification (CepA, CblA, CfxA, CSP-1 and TEM), class B (CfiA) or class D in Fusobacterium nucleatum (FUS-1). The minimum inhibitory concentrations of β-lactams are variable and this variation is often related to the presence of plasmids or other mobile genetic elements (MGEs) that modulate the expression of resistance genes. DNA persistence and bacterial promiscuity in oral biofilms also contribute to genetic transformation and conjugation in this particular microcosm. Overexpression of efflux pumps is facilitated because the encoding genes are located on MGEs, in some multidrug-resistant clinical isolates, similar to conjugative transposons harbouring genes encoding β-lactamases. All these facts lead us to consider the oral cavity as an important reservoir of β-lactam resistance genes and a privileged place for genetic exchange, especially in commensal strictly anaerobic Gram-negative bacilli. Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  4. Effect of intestinal microflora on the survival time of mice exposed to lethal whole-body. gamma. irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Onoue, M.; Uchida, K.; Yokokura, T.; Takahashi, T.; Mutai, M.

    1981-11-01

    The effect of intestinal microflora on the survival time of mice exposed to 2-kR whole-body ..gamma.. irradiation was studied using germfree, monoassociated, and conventionalized ICR mice. The germfree mice were monoassociated with 1 of 11 bacterial strains, which were isolated from the fresh feces of conventional mice, 2 weeks prior to irradiation. All mice died within 3 weeks after irradiation. Monoassociation with Fusobacterium sp., Streptococcus faecalis, Escherichia coli, or Pseudomonas sp. significantly reduced the mean survival time compared to that of germfree mice. In contrast, monoassociation with Clostridium sp., Bifidobacterium pseudolongum, or Lactobacillus acidophilus significantly prolonged the mean survival time compared to that of germfree mice. This suggests that the latter organisms may perform some activity to protect the mice from radiation injury. In this histopathological autopsy examination, the main lesions were hypocellularity in hematopoietic organs and hemorrhage in various organs. Neither karyorrhexis nor desquamation of intestinal mucosal cells was observed in any mice. From these observations, it is suggested that the death of these mice was related to hematopoietic damage. Bacterial invasion into various organs was observed in conventionalized and Pseudomonas-, E. coli-, or S. faecalis-monoassociated mice but not in Clostridium-, B. pseudolongum-, L. acidophilus-, or Fusobacterium-monoassociated mice.

  5. Effect of intestinal microflora on the survival time of mice exposed to lethal whole-body γ irradiation

    International Nuclear Information System (INIS)

    Onoue, M.; Uchida, K.; Yokokura, T.; Takahashi, T.; Mutai, M.

    1981-01-01

    The effect of intestinal microflora on the survival time of mice exposed to 2-kR whole-body γ irradiation was studied using germfree, monoassociated, and conventionalized ICR mice. The germfree mice were monoassociated with 1 of 11 bacterial strains, which were isolated from the fresh feces of conventional mice, 2 weeks prior to irradiation. All mice died within 3 weeks after irradiation. Monoassociation with Fusobacterium sp., Streptococcus faecalis, Escherichia coli, or Pseudomonas sp. significantly reduced the mean survival time compared to that of germfree mice. In contrast, monoassociation with Clostridium sp., Bifidobacterium pseudolongum, or Lactobacillus acidophilus significantly prolonged the mean survival time compared to that of germfree mice. This suggests that the latter organisms may perform some activity to protect the mice from radiation injury. In this histopathological autopsy examination, the main lesions were hypocellularity in hematopoietic organs and hemorrhage in various organs. Neither karyorrhexis nor desquamation of intestinal mucosal cells was observed in any mice. From these observations, it is suggested that the death of these mice was related to hematopoietic damage. Bacterial invasion into various organs was observed in conventionalized and Pseudomonas-, E. coli-, or S. faecalis-monoassociated mice but not in Clostridium-, B. pseudolongum-, L. acidophilus-, or Fusobacterium-monoassociated mice

  6. Synergic activity, for anaerobes, of trovafloxacin with clindamycin or metronidazole: chequerboard and time-kill methods.

    Science.gov (United States)

    Ednie, L M; Credito, K L; Khantipong, M; Jacobs, M R; Appelbaum, P C

    2000-05-01

    Chequerboard titrations were used to test the activity of trovafloxacin, alone and in combination with clindamycin or metronidazole, against 156 Gram-positive or Gram-negative anaerobes, including 47 Bacteroides fragilis group, 36 Prevotella spp., 26 fusobacteria, 21 peptostreptococci and 26 clostridia. MIC50/MIC90 values (mg/L) of each drug alone against all 156 strains were: trovafloxacin, 0.5/1; clindamycin, 0.25/2; metronidazole, 1/2. Synergy (FIC indices 0. 5-2.0); no antagonism (FIC indices >4.0) was seen. In addition, synergy was tested by time-kill methodology for each of the above combinations against 12 Gram-positive or Gram-negative strains. Results indicated that synergy (defined as a >/= 2 log(10) decrease in cfu/mL at 48 h compared with the more active drug alone) was found between trovafloxacin at or below the MIC and both clindamycin and metronidazole at or below the MIC in one strain each of Bacteroides fragilis, Bacteroides thetaiotaomicron, Prevotella intermedia, Fusobacterium varium, Peptostreptococcus asaccharolyticus and Clostridium bifermentans. Synergy between trovafloxacin (Fusobacterium mortiferum, P. asaccharolyticus and C. bifermentans. In many cases of synergy, including those at the trovafloxacin MIC, regrowth after 48 h, which was commonly seen with trovafloxacin alone, was inhibited, and 99.9% killing was observed with the combination after 48 h, but not with trovafloxacin alone.

  7. Composition and abundance of microbiota in the pharynx in patients with laryngeal carcinoma and vocal cord polyps.

    Science.gov (United States)

    Gong, Hongli; Wang, Boyan; Shi, Yi; Shi, Yong; Xiao, Xiyan; Cao, Pengyu; Tao, Lei; Wang, Yuezhu; Zhou, Liang

    2017-08-01

    The pharynx is an important site of microbiota colonization, but the bacterial populations at this site have been relatively unexplored by culture-independent approaches. The aim of this study was to characterize the microbiota structure of the pharynx. Pyrosequencing of 16S rRNA gene libraries was used to characterize the pharyngeal microbiota using swab samples from 68 subjects with laryngeal cancer and 28 subjects with vocal cord polyps. Overall, the major phylum was Firmicutes, with Streptococcus as the predominant genus in the pharyngeal communities. Nine core operational taxonomic units detected from Streptococcus, Fusobacterium, Prevotella, Granulicatella, and Veillonella accounted for 21.3% of the total sequences detected. However, there was no difference in bacterial communities in the pharynx from patients with laryngeal cancer and vocal cord polyps. The relative abundance of Firmicutes was inversely correlated with Fusobacteria, Proteobacteria, Actinobacteria, and Bacteroidetes. The correlation was evident at the genus level, and the relative abundance of Streptococcus was inversely associated with Fusobacterium, Leptotrichia, Neisseria, Actinomyces, and Prevotella. This study presented a profile for the overall structure of the microbiota in pharyngeal swab samples. Inverse correlations were found between Streptococcus and other bacterial communities, suggesting that potential antagonism may exist among pharyngeal microbiota.

  8. Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter

    International Nuclear Information System (INIS)

    Pszon-Bartosz, Kamila; Hansen, Jesper S.; Stibius, Karin B.; Groth, Jesper S.; Emneus, Jenny; Geschke, Oliver; Helix-Nielsen, Claus

    2011-01-01

    Research highlights: → We have established a vesicle fusion efficacy assay based on the major non-specific porin of Fusobacterium nucleatum (FomA). → Maximal fusion obtained was almost 150,000 porin insertions during 20 min. → Incorporation can be either first order or exponential kinetics which has implications for establishing protein delivery to biomimetic membranes. -- Abstract: Reconstitution of functionally active membrane protein into artificially made lipid bilayers is a challenge that must be overcome to create a membrane-based biomimetic sensor and separation device. In this study we address the efficacy of proteoliposome fusion with planar membrane arrays. We establish a protein incorporation efficacy assay using the major non-specific porin of Fusobacterium nucleatum (FomA) as reporter. We use electrical conductance measurements and fluorescence microscopy to characterize proteoliposome fusion with an array of planar membranes. We show that protein reconstitution in biomimetic membrane arrays may be quantified using the developed FomA assay. Specifically, we show that FomA vesicles are inherently fusigenic. Optimal FomA incorporation is obtained with a proteoliposome lipid-to-protein molar ratio (LPR) = 50 more than 10 5 FomA proteins could be incorporated in a bilayer array with a total membrane area of 2 mm 2 within 20 min. This novel assay for quantifying protein delivery into lipid bilayers may be a useful tool in developing biomimetic membrane applications.

  9. Isolation of non-sporing anaerobic rods from infections in children.

    Science.gov (United States)

    Brook, I

    1996-07-01

    From 1974 to 1994, 2033 microbiological specimens from children were submitted for cultures for anaerobic bacteria. Fifty-seven isolates of Bifidobacterium spp. were obtained from 55 (3%) children, 67 isolates of Eubacterium spp. from 65 (3%) children and 41 isolates of Lactobacillus spp. from 40 (2%) children. Most Bifidobacterium isolates were from chronic otitis media, abscesses, peritonitis, aspiration pneumonia and paronychia. Most Eubacterium isolates were from abscesses, peritonitis, decubitus ulcers and bites. Lactobacillus spp. were mainly isolated from abscesses, aspiration pneumonia, bacteraemia and conjunctivitis. Most (> 90%) infections from which these species were isolated were polymicrobial and yielded a mixture of aerobic and anaerobic bacteria. The organisms most commonly isolated with the non-sporing anaerobic gram-positive rods were Peptostreptococcus spp., Bacteroides spp., pigmented Prevotella and Porphyromonas spp., Fusobacterium spp., Staphylococcus aureus and Escherichia coli. Most Bacteroides spp. and E. coli were isolated from intra-abdominal infection and skin and soft tissue infection around the rectal area, whereas most Prevotella, Porphyromonas and Fusobacterium isolates were from oropharyngeal, pulmonary and head and neck sites. The predisposing conditions associated with the isolation of non-sporing anaerobic gram-positive rods were previous surgery, malignancy, steroid therapy and immunodeficiency. Antimicrobial therapy was given to 149 (83%) of the 160 patients, in conjunction with surgical drainage or correction of pathology in 89 (56%).

  10. Evaluation of the RapID-ANA system for identification of anaerobic bacteria of veterinary origin.

    Science.gov (United States)

    Adney, W S; Jones, R L

    1985-12-01

    This study evaluated the ability of the RapID-ANA system (Innovative Diagnostic Systems, Inc., Atlanta, Ga.) to accurately identify a spectrum of freshly isolated veterinary anaerobes. A total of 183 isolates were tested and included 7 Actinomyces spp., 53 Bacteroides spp., 32 Clostridium spp., 2 Eubacterium spp., 65 Fusobacterium spp., 1 Peptococcus spp., 22 Peptostreptococcus spp., and 1 Propionibacterium spp. All isolates were initially identified by conventional biochemical testing and gas-liquid chromatography of short-chain fatty acid metabolites. Additional tests were performed as required by the RapID-ANA system. Of these isolates, 81.4% were correctly identified to the genus level, including 59.6% to the species level, 14.2% were incorrectly identified at the genus level, and 4.4% were not identified. Initially, 20.2% of the strains were not identified because the microcodes were not in the code book. The majority of the incorrect identifications were caused by the misidentification of Fusobacterium spp. as Bacteroides spp. Errors also occurred when veterinary anaerobes not included in the data base were assigned an identification from the existing data base. The RapID-ANA system appears to be a promising new method for rapid identification of veterinary anaerobes; however, further evaluation with an extended data base is needed before the system can accurately identify all clinically significant anaerobes.

  11. An unusual autopsy case of pyogenic liver abscess caused by periodontal bacteria.

    Science.gov (United States)

    Ohyama, Hideki; Nakasho, Keiji; Yamanegi, Koji; Noiri, Yuichiro; Kuhara, Ayako; Kato-Kogoe, Nahoko; Yamada, Naoko; Hata, Masaki; Nishimura, Fusanori; Ebisu, Shigeyuki; Terada, Nobuyuki

    2009-09-01

    Pyogenic liver abscess (PLA) formation is thought to originate from the transmission of infection via three major routes including the biliary tract, portal vein and hepatic artery. However, about 50% of PLA cases are considered to be cryptogenic. Here we report an unusual autopsy case of PLA associated with periodontopathic bacterial infection. A 59-year-old female suddenly developed cardiopulmonary arrest and died. Despite macroscopic and microscopic examinations, the infectious routes and source of infection were unidentified, and the case appeared to be cryptogenic. Since this patient had suffered severe periodontitis for a long period of time, we investigated the involvement of periodontal infection in PLA formation by performing immunohistochemical analyses. We identified several periodontopathic bacterial species in the PLA of this patient, including Fusobacterium nucleatum, Treponema denticola, Prevotella intermedia and Porphyromonas gingivalis. Thus, we demonstrate here that periodontal infection is a potential source of infection in the formation of PLA.

  12. NÖTR VE ALKALİ pH'LARDAKİ KALSİYUM HİDROKSİT VE TİTANYUM DİOKSİT PATLARININ ANTİMİKROP ETKİLERİNİN KARŞILAŞTIRMALI OLARAK İNCELENMESİ*-COMPARISON OF ANTIBACTERIAL EFFECTIVENESS OF TITANIUM DIOXIDE AND CALCIUM HYDROXIDE PASTES AT NEUTRAL AND ALKALINE pHs

    OpenAIRE

    Dindar, Seçkin; Külekçi, Güven; Özbaş, Hakan; Turan, Fatma

    2013-01-01

    ÖzetBu in vitro çalışmada farklı pH'lardaki titanyum diok-sit (Tİ02) ve kalsiyum hidroksit (Ca (OH)2)'nin anti-mikrop etkilerinin araştırılması amaçlanmıştır. TİO2 ve Ca (OH)2 patları pH:7 ve pH:10 olmak üzere hazır­lanmıştır. Patların antimikrop etkileri Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas fluorescens, Escherichia coli, Klebsiella pneumoniae, Peptostreptococcus sp., Lactobacillus sp., Fusobacte-rium sp., Porphyromonas gingivalis, Bacteroides fragilis, Clo...

  13. NÖTR VE ALKALİ pH'LARDAKİ KALSİYUM HİDROKSİT VE TİTANYUM DİOKSİT PATLARININ ANTİMİKROP ETKİLERİNİN KARŞILAŞTIRMALI OLARAK İNCELENMESİ*-COMPARISON OF ANTIBACTERIAL EFFECTIVENESS OF TITANIUM DIOXIDE AND CALCIUM HYDROXIDE PASTES AT NEUTRAL AND ALKALINE pHs

    OpenAIRE

    Dindar, Seçkin; Külekçi, Güven; Özbaş, Hakan; Turan, Fatma

    2013-01-01

    ÖzetBu in vitro çalışmada farklı pH'lardaki titanyum diok-sit (Tİ02) ve kalsiyum hidroksit (Ca (OH)2)'nin anti-mikrop etkilerinin araştırılması amaçlanmıştır. TİO2 ve Ca (OH)2 patları pH:7 ve pH:10 olmak üzere hazır­lanmıştır. Patların antimikrop etkileri Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas fluorescens, Escherichia coli, Klebsiella pneumoniae, Peptostreptococcus sp., Lactobacillus sp., Fusobacte-rium sp., Porphyromonas gingivalis, Bacteroides fragilis, Clostridium s...

  14. [Effect of compound Chinese traditional medicine on infected root canal bacteria biofilm].

    Science.gov (United States)

    Ma, Rui; Huang, Li-li; Xia, Wen-wei; Zhu, Cai-lian; Ye, Dong-xia

    2010-08-01

    To assess the efficacy of compound Chinese traditional medicine(CTM), which composed of gallic acid, magnolol and polysaccharide of Blettila striata, against the infected root canal bacterial biofilm. Actinomyces viscosus (Av), Enterococcus faecalis (Ef), Fusobacterium nucleatum (Fn) were composed to form biofilm, then confocal laser scan microscope (CLSM) was used to observe and study the bacterial activity. SAS6.12 software package was used for statistical analysis. The biofilm thickness reduced after treatment by both CTM and ZnO (P>0.05),while there was a significant decrease of the percentage of vital bacterias after treatment by CTM (Pcompound Chinese traditional medicine is effective on biofilm control, so that it would be an effective disinfecting drug for root canal sealers. Supported by Research Fund of Bureau of Traditional Chinese Medicine of Shanghai Municipality (Grant No.2008L008A).

  15. Probiotic Lactobacillus rhamnosus GG prevents alveolar bone loss in a mouse model of experimental periodontitis.

    Science.gov (United States)

    Gatej, Simona M; Marino, Victor; Bright, Richard; Fitzsimmons, Tracy R; Gully, Neville; Zilm, Peter; Gibson, Rachel J; Edwards, Suzanne; Bartold, Peter M

    2018-02-01

    This study investigated the role of Lactobacillus rhamnosus GG (LGG) on bone loss and local and systemic inflammation in an in vivo mouse model of experimental periodontitis (PD). Experimental PD was induced in mice by oral inoculation with Porphyromonas gingivalis and Fusobacterium nucleatum over a period of 44 days. The probiotic LGG was administered via oral inoculation or oral gavage prior to, and during disease induction. The antimicrobial activity of LGG on the inoculum was also tested. Alveolar bone levels and gingival tissue changes were assessed using in vivo microcomputed tomography and histological analysis. Serum levels of mouse homologues for IL-8 were measured using multiplex assays. Pre-treatment with probiotics either via oral gavage or via oral inoculation significantly reduced bone loss (p loss in a mouse model of induced PD irrespective of the mode of administration. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  16. Assessing the efficacy of vesicle fusion with planar membrane arrays using a mitochondrial porin as reporter

    DEFF Research Database (Denmark)

    Pszon-Bartosz, Kamila Justyna; Hansen, Jesper S.; Stibius, Karin B.

    2011-01-01

    Reconstitution of functionally active membrane protein into artificially made lipid bilayers is a challenge that must be overcome to create a membrane-based biomimetic sensor and separation device. In this study we address the efficacy of proteoliposome fusion with planar membrane arrays. We...... establish a protein incorporation efficacy assay using the major non-specific porin of Fusobacterium nucleatum (FomA) as reporter. We use electrical conductance measurements and fluorescence microscopy to characterize proteoliposome fusion with an array of planar membranes. We show that protein...... reconstitution in biomimetic membrane arrays may be quantified using the developed FomA assay. Specifically, we show that FomA vesicles are inherently fusigenic. Optimal FomA incorporation is obtained with a proteoliposome lipid-to-protein molar ratio (LPR)=50 more than 105 FomA proteins could be incorporated...

  17. Anaerobes in pleuropulmonary infections

    Directory of Open Access Journals (Sweden)

    De A

    2002-01-01

    Full Text Available A total of 76 anaerobes and 122 aerobes were isolated from 100 patients with pleuropulmonary infections, e.g. empyema (64, pleural effusion (19 and lung abscess (13. In 14% of the patients, only anaerobes were recovered, while a mixture of aerobes and anaerobes was encountered in 58%. From all cases of lung abscess, anaerobic bacteria were isolated, alone (04 or along with aerobic bacteria (13. From empyema and pleural effusion cases, 65.6% and 68.4% anaerobes were recovered respectively. Amongst anaerobes, gram negative anaerobic bacilli predominated (Prevotella melaninogenicus 16, Fusobacterium spp. 10, Bacteroides spp. 9, followed by gram positive anaerobic cocci (Peptostreptococcus spp. 31. Coliform bacteria (45 and Pseudomonas aeruginosa (42 were the predominant aerobic isolates.

  18. Antibacterial efficacy of triple-layered poly(lactic-co-glycolic acid)/nanoapatite/lauric acid guided bone regeneration membrane on periodontal bacteria.

    Science.gov (United States)

    Saarani, Nur Najiha; Jamuna-Thevi, Kalitheerta; Shahab, Neelam; Hermawan, Hendra; Saidin, Syafiqah

    2017-05-31

    A guided bone regeneration (GBR) membrane has been extensively used in the repair and regeneration of damaged periodontal tissues. One of the main challenges of GBR restoration is bacterial colonization on the membrane, constitutes to premature membrane degradation. Therefore, the purpose of this study was to investigate the antibacterial efficacy of triple-layered GBR membrane composed of poly(lactic-co-glycolic acid) (PLGA), nanoapatite (NAp) and lauric acid (LA) with two types of Gram-negative periodontal bacteria, Fusobacterium nucleatum and Porphyromonas gingivalis through a disc diffusion and bacterial count tests. The membranes exhibited a pattern of growth inhibition and killing effect against both bacteria. The increase in LA concentration tended to increase the bactericidal activities which indicated by higher diameter of inhibition zone and higher antibacterial percentage. It is shown that the incorporation of LA into the GBR membrane has retarded the growth and proliferation of Gram-negative periodontal bacteria for the treatment of periodontal disease.

  19. [Significance of the bacterial flora in the etiology of apical periodontitis. Qualitative, quantitative and topographical aspects].

    Science.gov (United States)

    Accorsi, S; Lavagnoli, G; Frigeri, S; Fiamminghi, L

    1990-01-01

    In the international literature it is shown the central role of root canal infection in the etiology of periapical lesions. As a matter of fact it has been proved (13) that sterile necrotic pulp tissue is completely unable to cause inflammatory reactions at the periapex. Infection of endodontic origin extends to the supporting tissues of the tooth only in the case of their acute inflammation (e.g. acute apical periodontitis, acute alveolar abscess, phoenix abscess). On the other hand in chronic inflammation bacteria remain confined in the endodontic space. Only few exceptions to this general rule have been experimentally proved. In endodontics we deal with a mixed infection which is composed by obligate anaerobes and by facultative anaerobes. The most frequently found obligate anaerobes are Bacteroides sp. and Fusobacterium sp. (Gram- rods) Anaerobic Diphtheroides (Gram+ rods) Peptostreptococcus sp. (Gram+ cocci) and Veilonella sp. (Gram- cocci). Actinomyces sp., Lactobacillus sp., Streptococcus sp., and Staphilococcus sp. are the facultative anaerobes most frequently found.

  20. Bi-Mix Antimicrobial Scaffolds for Regenerative Endodontics

    Science.gov (United States)

    Palasuk, Jadesada; Kamocki, Krzysztof; Hippenmeyer, Lauren; Platt, Jeffrey A.; Spolnik, Kenneth J.; Gregory, Richard L.; Bottino, Marco C.

    2014-01-01

    Introduction Eliminating and/or inhibiting bacterial growth within the root canal system have been shown to play a key role in the regenerative outcome. The aim of this study was to synthesize and determine in vitro both the antimicrobial effectiveness and cytocompatibility of bi-mix antibiotic-containing polydioxanone (PDS)-based polymer scaffolds. Methods Antibiotic-containing (metronidazole, MET and ciprofloxacin, CIP) polymer solutions (distinct antibiotic weight ratios) were spun into fibers as a potential mimic to the double antibiotic paste (DAP, a MET/CIP mixture). Fiber morphology, chemical characteristics, and tensile strength were evaluated by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and tensile testing, respectively. Antimicrobial efficacy was tested over time (aliquot collection) against Enterococcus faecalis (Ef), Porphyromonas gingivalis (Pg), and Fusobacterium nucleatum (Fn). Similarly, cytotoxicity was evaluated in human dental pulp stem cells (hDPSCs). Data were statistically analyzed (pendodontics. PMID:25201643

  1. Antimicrobial Activity of Diterpenes from Viguiera arenaria against Endodontic Bacteria

    Directory of Open Access Journals (Sweden)

    Carlos H. G. Martins

    2011-01-01

    Full Text Available Six pimarane-type diterpenes isolated from Viguiera arenaria Baker and two semi-synthetic derivatives were evaluated in vitro against a panel of representative microorganisms responsible for dental root canal infections. The microdilution method was used for the determination of the minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC against Porphyromonas gingivalis, Prevotella nigrescens, Prevotella intermedia, Prevotella buccae, Fusobacterium nucleatum, Bacteroides fragilis, Actinomyces naeslundii, Actinomyces viscosus, Peptostreptococcus micros, Enterococcus faecalis and Aggregatibacter actinomycetemcomitans. The compounds ent-pimara-8(14,15-dien-19-oic acid, its sodium salt and ent-8(14,15-pimaradien-3β-ol were the most active, displaying MIC values ranging from 1 to 10 μg mL-1. The results also allow us to conclude that minor structural differences among these diterpenes significantly influence their antimicrobial activity, bringing new perspectives to the discovery of new chemicals for use as a complement to instrumental endodontic procedures.

  2. In vitro cytokine responses to periodontal pathogens: generalized aggressive periodontitis is associated with increased IL-6 response to Porphyromonas gingivalis

    DEFF Research Database (Denmark)

    Borch, T S; Holmstrup, Palle; Bendtzen, K

    2010-01-01

    with GAgP and 10 controls stimulated with periodontal pathogens or a control antigen, tetanus toxoid (TT) in the presence of autologous serum. The pathogens used were Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum, either as type strains or bacteria isolated from...... the participants' inherent oral flora. The P. gingivalis -induced production of IL-6 was approximately 2.5-fold higher in patients with GAgP than in healthy controls (P gingivalis, as all cytokine...... responses induced by Pr. intermedia, F. nucleatum and TT was similar in the two groups. A reduced IL-12p70 response to Pr. intermedia and F. nucleatum was observed in smokers compared to non-smoking patients (P gingivalis, MNC...

  3. ASSOCIATION OF THE BONE LOSS WITH MAIN CLINICAL AND MICROBIOLOGICAL PARAMETERS IN CHRONIC PERIODONTITIS

    Directory of Open Access Journals (Sweden)

    Velitchka Dosseva-Panova

    2014-07-01

    Full Text Available Literature data have reported that certain Gram-negative anaerobe bacteria from the subgingival environment are important etiological agents of chronic periodontal disease. Based on published criteria identifying periodontal pathogens, major evidences has been shown for: Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia; moderate evidences for: Prevotella intermedia, Peptostreptococcus /Micromonas/ micros, Fusobacterium nucleatum, Eubacterium nodatum, Campylobacter rectus, and spirochetes (Treponema denticola. Microbiological data has been suggested by several studies as a useful indicator of disease progression, selection of an adjunctive antimicrobial administration, healing assessment and disease recurrence. The results of this study demonstrate statistically significant correlation between periodontal attachment loss and the prevalence of Porphyromonas gingivalis, Tannerella forsythia and Peptostreptococcus (Micromonas micros in deep periodontal pockets of severe chronic periodontitis patients.

  4. CLINICAL AND MICROBIOLOGICAL DATA IN PATIENTS WITH CHRONIC PERIODONTITIS.

    Directory of Open Access Journals (Sweden)

    Christina Popova

    2013-07-01

    Full Text Available Periodontitis is an infectious disease concerning supporting tissues of the teeth. The primary etiological agent for disease development and progression is the subgingival biofilm, but recently it is known that host factors may modify the pathological process or may affect the severity and /or extent. The increasing levels of some specific pathogenic subgingival bacteria such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia and others can result in periodontal destruction and possibly correlate with disease severity. Data from controlled studies show high prevalence of P. gingivalis, T. forsythia and Tr. denticola which represent the red complex (coexistence of these three species in patients with moderate and severe chronic periodontitis. Parallel investigation of probing depth (PD and clinical attachment level (CAL with the microbiological testing may give a confirmation of relation between subgingival pathogenic bacteria and severity of periodontitis.

  5. Association between periodontal condition and subgingival microbiota in women during pregnancy: a longitudinal study

    Directory of Open Access Journals (Sweden)

    Priscila Viola BORGO

    2014-12-01

    Full Text Available Objectivo In this study, the gingival conditions and the quantitative detection for Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in pregnant women were determined. Material and Methods Quantitative determinations of periodontal bacteria by using a SyBr green system in women during pregnancy were performed. Women at the 2nd and 3rd trimesters of pregnancy and non-pregnant women were included in this study. A. actinomycetemcomitans was observed in high numbers in women at the 2nd and 3rd trimesters of pregnancy with a significant difference (p<0.05. F. nucleatum and P. intermedia were also observed in high levels. Results and Conclusion Our results show that pregnant women are more susceptible to gingivitis, and the presence of A. actinomycetemcomitans in subgingival biofilm might be taken into account for the treatment of periodontal disease.

  6. Altered Bacterial Profiles in Saliva from Adults with Caries Lesions

    DEFF Research Database (Denmark)

    Belstrøm, D; Fiehn, N-E; Nielsen, C H

    2014-01-01

    -Whitney's test with Benjamini-Hochberg correction for multiple comparisons. Principal component analysis was used to visualize bacterial community profiles. A reduced bacterial diversity was observed in samples from subjects with dental caries. Five bacterial taxa (Veillonella parvula, Veillonella atypica......, Megasphaera micronuciformis, Fusobacterium periodontium and Achromobacter xylosoxidans) and one bacterial cluster (Leptotrichia sp. clones C3MKM102 and GT018_ot417/462) were less frequently found in the caries group (adjusted p value ... salivarius) and three bacterial clusters (Streptococcus parasanguinis I and II and sp. clone BE024_ot057/411/721, Streptococcus parasanguinis I and II and sinensis_ot411/721/767, Streptococcus salivarius and sp. clone FO042_ot067/755) were present at significantly higher levels (adjusted p value

  7. Activity of endodontic antibacterial agents against selected anaerobic bacteria

    Directory of Open Access Journals (Sweden)

    Ferreira Cláudio Maniglia

    2002-01-01

    Full Text Available The antimicrobial activity of substances used as antibacterial agents (solutions of 10% calcium hydroxide, camphorated paramonochlorophenol - PMCC, 2% chlorhexidine digluconate and 10% castor oil plant detergent on anaerobic bacteria (Fusobacterium nucleatum ATCC 25586, Prevotella nigrescens ATCC 33563, Clostridium perfringens ATCC 13124 and Bacteroides fragilis ATCC 25285, using a broth dilution technique, was evaluated in vitro. For determination of minimum inhibitory and minimum bactericide concentrations (MIC and MBC, two culture broths, Reinforced Clostridial Medium (RCM and supplemented Brucella, standardized inoculum and serially diluted solutions were used. All antibacterial agents presented antimicrobial activity that varied for different bacteria. There were no differences in the performance of the two broths. Chlorhexidine digluconate was the most effective, with the lowest MICs, followed by castor oil detergent, PMCC and calcium hydroxide. C. perfringens and B. fragilis were the most resistant bacteria to all agents.

  8. Inhibitory effect of Lactobacillus reuteri on periodontopathic and cariogenic bacteria.

    Science.gov (United States)

    Kang, Mi-Sun; Oh, Jong-Suk; Lee, Hyun-Chul; Lim, Hoi-Soon; Lee, Seok-Woo; Yang, Kyu-Ho; Choi, Nam-Ki; Kim, Seon-Mi

    2011-04-01

    The interaction between Lactobacillus reuteri, a probiotic bacterium, and oral pathogenic bacteria have not been studied adequately. This study examined the effects of L. reuteri on the proliferation of periodontopathic bacteria including Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, and Tannerella forsythia, and on the formation of Streptococcus mutans biofilms. Human-derived L. reuteri strains (KCTC 3594 and KCTC 3678) and rat-derived L. reuteri KCTC 3679 were used. All strains exhibited significant inhibitory effects on the growth of periodontopathic bacteria and the formation of S. mutans biofilms. These antibacterial activities of L. reuteri were attributed to the production of organic acids, hydrogen peroxide, and a bacteriocin-like compound. Reuterin, an antimicrobial factor, was produced only by L. reuteri KCTC 3594. In addition, L. reuteri inhibited the production of methyl mercaptan by F. nucleatum and P. gingivalis. Overall, these results suggest that L. reuteri may be useful as a probiotic agent for improving oral health.

  9. PCR-based identification of selected pathogens associated with endodontic infections in deciduous and permanent teeth.

    Science.gov (United States)

    Cogulu, Dilsah; Uzel, Atac; Oncag, Ozant; Eronat, Cemal

    2008-09-01

    The aim of the present study was to evaluate the presence of the selected pathogens in samples from deciduous and permanent tooth root canals by using PCR method and to determine the association of these organisms with clinical symptoms. A total of 145 children, 5 to 13 years old, were involved in this study. The presence of selected pathogens (Actinomyces israelii, Candida albicans, Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Streptococcus intermedius, Treponema denticola, Parvimonas micra, Tannerella forsythensis, Enterococcus faecium, Prevotella melaninogenica) in infected root canals was studied using PCR. T. denticola (P = .012, .02) and E. faecalis (P = .012, .04) were highly associated with periapical radiolucency and previous pain, while P. gingivalis was associated with tenderness to percussion in both deciduous and permanent teeth (P = .01, .015). The results of the present study confirm that certain species of microorganisms are associated with clinical signs and symptoms of endodontic disease in both deciduous and permanent teeth.

  10. Characteristics of enriched cultures for bio-huff-`n`-puff tests at Jilin oil field

    Energy Technology Data Exchange (ETDEWEB)

    Xiu-Yuan Wang; Gang Dai; Yan-Fen Xue; Shu-Hua Xie [Institute of Microbiology, Beijing (China)] [and others

    1995-12-31

    Three enriched cultures (48, 15a, and 26a), selected from more than 80 soil and water samples, could grow anaerobically in the presence of crude oil at 30{degrees}C and could ferment molasses to gases and organic acids. Oil recovery by culture 48 in the laboratory model experiment was enhanced by 25.2% over the original reserves and by 53.7% over the residual reserves. Enriched culture 48 was composed of at least 4 species belonging to the genera Eubacterium, Fusobacterium, and Bacteroides. This enriched culture was used as inoculum for MEOR field trials at Jilin oil field with satisfactory results. The importance of the role of these isolates in EOR was confirmed by their presence and behavior in the fluids produced from the microbiologically treated reservoir.

  11. [Comparative studying of anaerobic bacteria located in woman's reproductive ways in normal condition and dysbiosis].

    Science.gov (United States)

    Polishko, T N; Sirokvasha, E A; Klokov, V V; Vinnikov, A I

    2008-01-01

    Bacteriological investigation of obligate anaerobic bacteria located in UGT of two groups of the observed women has shown: that the microbiocoenosis of UGT of women of the group 1 can be determined as normal. Identification of these anaerobic bacteria revealed the presence of representatives of the following species: Lactobacillus spp., Bifidobacterium spp., Eubacterium spp., Bacteroides spp., Fusobacterium spp., Peptococcus spp., Peptostreptococcus spp. The microbiocoenosis of UGT of the women of group 2 is diagnosed as vaginosis, thus in addition to the listed previously bacteria is added another one, Clostridium spp. Characteristic feature of Vaginosis is from one side a considerable decrease in the frequency of finding (cultivation) and concentration of Lactobacillus spp. and Bifidobacterium spp. and from another side--a considerable increase of frequency finding (cultivation) and concentration of Bacteroides spp. In addition, there is change of metabolism of Lactobacillus spp. and Bifidobacterium spp resulting in decrease in specific intensity of secretion of acids.

  12. Positive and negative associations between bacterial species in dental root canals.

    Science.gov (United States)

    Gomes, B P; Drucker, D B; Lilley, J D

    1994-01-01

    Significant associations have been previously reported between certain pairs of bacterial species isolated from human dental root canals. The aim of this study was to examine microbiologically a more extensive series of cases, with particular reference to obligate anaerobes which accounted for 64% of total isolations. A total of 65 different species was isolated and individual root canals yielded a maximum of eleven bacterial species. Highly significant positive associations (p spp. and Prevotella spp., between Peptostreptococcus spp. and P. melaninogenica, between P. micros and Prevotella spp., P. micros and P. melaninogenica and between Prevotella spp. and Eubacterium spp., all with an ODDS ratio of > 9.0. In contrast, negative and highly significant associations (p spp., B. gracilis/F. nucleatum and between B. gracilis/Fusobacterium spp.; all with an ODDS ratio of < 0.5. Some previously published associations were confirmed and some new associations were found, while some negative associations became apparent.

  13. [Isolation of anaerobes during a 30-month observation at a hospital microbiology laboratory].

    Science.gov (United States)

    Pistono, P G; Rapetti, I; Stacchini, E; Vironda, N; D'Usi, M P; Guasco, C

    1989-01-01

    The authors evaluate retrospectively the results obtained from the research of anaerobial bacteria on 1313 samples received at the Microbiology Laboratory of the "Ospedale Civile di Ivrea" over a period of 31 months (6/1/86-12/31/88). From this evaluation, high percentages of detection of anaerobic bacteria are emerging in the following infections: appendiculare abscesses (60%), intestinal operations (71%), wounds (57%), tubovarian abscesses (100%), as well as thoracic empyema (50%). Also relevant are the isolations from skin and subcutaneous tissues: breast infections (50%) preputial infections (60%), perineal and perirectal abscesses (60%). The incident of anaerobic bacteria in bacteriemia is 17%. The most representative anaerobic bacteria group are: Bacteroides spp. (56%), Peptostreptococcus spp. (12%), Propionibacterium spp. (9%), Fusobacterium spp. (7%) Clostridium spp. (6%), Veillonella spp. and Eubacterium spp. (3%). In the intraabdominal infections prevails the Bacteroides group, particularly fragilis species, while in the skin and subcutaneous infections prevails the Peptostreptococcus group.

  14. Dysbiosis of oral buccal mucosa microbiota in patients with oral lichen planus.

    Science.gov (United States)

    He, Y; Gong, D; Shi, C; Shao, F; Shi, J; Fei, J

    2017-07-01

    The bacterial community structure of buccal mucosa in patients with oral lichen planus was evaluated and compared with healthy control. Buccal scraping samples have been taken on 43 oral lichen planus patients (21 erosive and 22 non-erosive) and 21 mucosal healthy volunteers. The V3 hypervariable 16S rDNA region was amplified and sequenced by high-throughput 454 pyrosequencing. 94.26% of the total buccal bacteria were classified into 15 abundant genera. Eight of these abundant genera could be detected in all cases, namely Streptococcus, Prevotella, Haemophilu, Neisseria, Fusobacterium, Leptotrichia, Veillonella and Actinomyces. Four abundant bacteria showed significantly different prevalence at the genus level: Streptococcus was more abundant (P oral microbiome. Further studies should be taken to elucidate the inner relationship between these observed changes and OLP development. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  15. Dental abscess: A microbiological review

    Directory of Open Access Journals (Sweden)

    Shweta

    2013-01-01

    Full Text Available Dental abscess is a frequently occurring infectious process known to the health practice. The fate of the infection depends on the virulence of the bacteria, host resistance factors, and regional anatomy. Serious consequences arising from the spread of a dental abscess lead to significant morbidity and mortality. Acute dental abscess is polymicrobial, comprising of strict anaerobes, such as anaerobic cocci, Prevotella, Fusobacterium species, and facultative anaerobes, such as viridans group streptococci and the Streptococcus anginosus group. Numerous novel, uncultivable and fastidious organisms have been identified as potential pathogens with the use of non-culture techniques. The majority of localized dental abscesses respond to surgical treatment while the use of antimicrobials is limited to severe spreading infections. There is a need for good-quality clinical trials of sufficient size to identify the ideal treatment. The microbiology of the acute dentoalveolar abscess and its treatment in the light of improved culture and diagnostic methods are reviewed.

  16. Lung Microbiota Is Related to Smoking Status and to Development of Acute Respiratory Distress Syndrome in Critically Ill Trauma Patients.

    Science.gov (United States)

    Panzer, Ariane R; Lynch, Susan V; Langelier, Chaz; Christie, Jason D; McCauley, Kathryn; Nelson, Mary; Cheung, Christopher K; Benowitz, Neal L; Cohen, Mitchell J; Calfee, Carolyn S

    2018-03-01

    Cigarette smoking is associated with increased risk of acute respiratory distress syndrome (ARDS) in patients after severe trauma; however, the mechanisms underlying this association are unknown. To determine whether cigarette smoking contributes to ARDS development after trauma by altering community composition of the lung microbiota. We studied the lung microbiota of mechanically ventilated patients admitted to the ICU after severe blunt trauma. To do so, we used 16S ribosomal RNA gene amplicon sequencing of endotracheal aspirate samples obtained on ICU admission (n = 74) and at 48 hours after admission (n = 30). Cigarette smoke exposure (quantified using plasma cotinine), ARDS development, and other clinical parameters were correlated with lung microbiota composition. Smoking status was significantly associated with lung bacterial community composition at ICU admission (P = 0.007 by permutational multivariate ANOVA [PERMANOVA]) and at 48 hours (P = 0.03 by PERMANOVA), as well as with significant enrichment of potential pathogens, including Streptococcus, Fusobacterium, Prevotella, Haemophilus, and Treponema. ARDS development was associated with lung community composition at 48 hours (P = 0.04 by PERMANOVA) and was characterized by relative enrichment of Enterobacteriaceae and of specific taxa enriched at baseline in smokers, including Prevotella and Fusobacterium. After severe blunt trauma, a history of smoking is related to lung microbiota composition, both at the time of ICU admission and at 48 hours. ARDS development is also correlated with respiratory microbial community structure at 48 hours and with taxa that are relatively enriched in smokers at ICU admission. The data derived from this pilot study suggest that smoking-related changes in the lung microbiota could be related to ARDS development after severe trauma.

  17. Comparative analysis of salivary bacterial microbiome diversity in edentulous infants and their mothers or primary care givers using pyrosequencing.

    Directory of Open Access Journals (Sweden)

    Kimberly D Cephas

    Full Text Available Bacterial contribution to oral disease has been studied in young children, but there is a lack of data addressing the developmental perspective in edentulous infants. Our primary objectives were to use pyrosequencing to phylogenetically characterize the salivary bacterial microbiome of edentulous infants and to make comparisons against their mothers. Saliva samples were collected from 5 edentulous infants (mean age = 4.6±1.2 mo old and their mothers or primary care givers (mean age = 30.8±9.5 y old. Salivary DNA was extracted, used to generate DNA amplicons of the V4-V6 hypervariable region of the bacterial 16S rDNA gene, and subjected to 454-pyrosequencing. On average, over 80,000 sequences per sample were generated. High bacterial diversity was noted in the saliva of adults [1012 operational taxonomical units (OTU at 3% divergence] and infants (578 OTU at 3% divergence. Firmicutes, Proteobacteria, Actinobacteria, and Fusobacteria were predominant bacterial phyla present in all samples. A total of 397 bacterial genera were present in our dataset. Of the 28 genera different (P<0.05 between infants and adults, 27 had a greater prevalence in adults. The exception was Streptococcus, which was the predominant genera in infant saliva (62.2% in infants vs. 20.4% in adults; P<0.05. Veillonella, Neisseria, Rothia, Haemophilus, Gemella, Granulicatella, Leptotrichia, and Fusobacterium were also predominant genera in infant samples, while Haemophilus, Neisseria, Veillonella, Fusobacterium, Oribacterium, Rothia, Treponema, and Actinomyces were predominant in adults. Our data demonstrate that although the adult saliva bacterial microbiome had a greater OTU count than infants, a rich bacterial community exists in the infant oral cavity prior to tooth eruption. Streptococcus, Veillonella, and Neisseria are the predominant bacterial genera present in infants. Further research is required to characterize the development of oral microbiota early in life

  18. Cervical Microbiome and Cytokine Profile at Various Stages of Cervical Cancer: A Pilot Study.

    Directory of Open Access Journals (Sweden)

    Astride Audirac-Chalifour

    Full Text Available Cervical cancer (CC is caused by high-risk human papillomavirus persistence due to the immunosuppressive tumor microenvironment mediated by cytokines. Vaginal microbiota determines the presence of certain cytokines locally. We assessed the association between cervical microbiota diversity and the histopathological diagnosis of each stage of CC, and we evaluated mRNA cervical expression levels of IL-4, IL-6, IL-10, TGF-β1, TNF-α and IFN-γ across the histopathological diagnosis and specific bacterial clusters. We determined the cervical microbiota by high throughput sequencing of 16S rDNA amplicons and classified it in community state types (CST. Mean difference analyses between alpha-diversity and histopathological diagnosis were carried out, as well as a β-diversity analysis within the histological diagnosis. Cervical cytokine mRNA expression was analyzed across the CSTs and the histopathological diagnoses. We found a significant difference in microbiota's diversity in NCL-HPV negative women vs those with squamous intraepithelial lesions (SIL and CC(p = 0.006, p = 0.036.When β-diversity was evaluated, the CC samples showed the highest variation within groups (p<0.0006 and the largest distance compared to NCL-HPV negative ones (p<0.00001. The predominant bacteria in women with normal cytology were L. crispatus and L. iners, whereas for SIL, it was Sneathia spp. and for CC, Fusobacterium spp. We found higher median cervical levels of IL-4 and TGF-β1 mRNA in the CST dominated by Fusobacterium spp. These results suggest that the cervical microbiota may be implicated in cervical cancer pathology. Further cohort studies are needed to validate these findings.

  19. In vitro efficacy of cefovecin against anaerobic bacteria isolated from subgingival plaque of dogs and cats with periodontal disease.

    Science.gov (United States)

    Khazandi, Manouchehr; Bird, Philip S; Owens, Jane; Wilson, Gary; Meyer, James N; Trott, Darren J

    2014-08-01

    Periodontal disease is a common disease of dogs and cats often requiring antimicrobial treatment as an adjunct to mechanical debridement. However, correct compliance with oral antimicrobial therapy in companion animals is often difficult. Cefovecin is a recently introduced veterinary cephalosporin that has demonstrated prolonged concentrations in extracellular fluid, allowing for dosing intervals of up to 14 days. Subgingival samples were collected from the oral cavity of 29 dogs and eight cats exhibiting grade 2 or grade 3 periodontal disease. Samples were cultivated on Wilkin Chalgrens agar and incubated in an anaerobic chamber for seven days. Selected anaerobic bacteria were isolated and identified to species level using 16S rRNA gene sequence analysis. Minimum inhibitory concentrations were determined for cefovecin and six additional antimicrobials using the agar dilution methodology recommended by the Clinical and Laboratory Standards Institute. The 65 clinical isolates were identified as Porphyromonas gulae (n = 45), Porphyromonas crevioricanis (n = 12), Porphyromonas macacae (n = 1), Porphyromonas cangingivalis (n = 1) Fusobacterium nucleatum (n = 2), Fusobacterium russii (n = 1) and Solobacterium moorei (n = 3). This is the first report of S. moorei being isolated from companion animals with periodontal disease. All isolates were highly susceptible to cefovecin, with a MIC90 of ≤0.125 μg/ml. Conversely, different resistance rates to ampicillin, amoxicillin and erythromycin between isolates were detected. Cefovecin is thus shown to be effective in vitro against anaerobic bacteria isolated from dogs and cats with periodontal disease. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Anaerobic co-culture of mesenchymal stem cells and anaerobic pathogens - a new in vitro model system.

    Directory of Open Access Journals (Sweden)

    Katja Kriebel

    Full Text Available BACKGROUND: Human mesenchymal stem cells (hMSCs are multipotent by nature and are originally isolated from bone marrow. In light of a future application of hMSCs in the oral cavity, a body compartment with varying oxygen partial pressures and an omnipresence of different bacterial species i.e. periodontitis pathogens, we performed this study to gain information about the behavior of hMSC in an anaerobic system and the response in interaction with oral bacterial pathogens. METHODOLOGY/PRINCIPAL FINDINGS: We established a model system with oral pathogenic bacterial species and eukaryotic cells cultured in anaerobic conditions. The facultative anaerobe bacteria Fusobacterium nucleatum, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were studied. Their effects on hMSCs and primary as well as permanent gingival epithelial cells (Ca9-22, HGPEC were comparatively analyzed. We show that hMSCs cope with anoxic conditions, since 40% vital cells remain after 72 h of anaerobic culture. The Ca9-22 and HGPEC cells are significantly more sensitive to lack of oxygen. All bacterial species reveal a comparatively low adherence to and internalization into hMSCs (0.2% and 0.01% of the initial inoculum, respectively. In comparison, the Ca9-22 and HGPEC cells present better targets for bacterial adherence and internalization. The production of the pro-inflammatory chemokine IL-8 is higher in both gingival epithelial cell lines compared to hMSCs and Fusobacterium nucleatum induce a time-dependent cytokine secretion in both cell lines. Porphyromonas gingivalis is less effective in stimulating secretion of IL-8 in the co-cultivation experiments. CONCLUSIONS/SIGNIFICANCE: HMSCs are suitable for use in anoxic regions of the oral cavity. The interaction with local pathogenic bacteria does not result in massive pro-inflammatory cytokine responses. The test system established in this study allowed further investigation of parameters prior to set up of

  1. PCR-based detection of resistance genes in anaerobic bacteria isolated from intra-abdominal infections.

    Science.gov (United States)

    Tran, Chau Minh; Tanaka, Kaori; Watanabe, Kunitomo

    2013-04-01

    Little information is available on the distribution of antimicrobial resistance genes in anaerobes in Japan. To understand the background of antimicrobial resistance in anaerobes involved in intra-abdominal infections, we investigated the distribution of eight antimicrobial resistance genes (cepA, cfiA, cfxA, ermF, ermB, mefA, tetQ, and nim) and a mutation in the gyrA gene in a total of 152 organisms (Bacteroides spp., Prevotella spp., Fusobacterium spp., Porphyromonas spp., Bilophila wadsworthia, Desulfovibrio desulfuricans, Veillonella spp., gram-positive cocci, and non-spore-forming gram-positive bacilli) isolated between 2003 and 2004 in Japan. The cepA gene was distributed primarily in Bacteroides fragilis. Gene cfxA was detected in about 9 % of the Bacteroides isolates and 75 % of the Prevotella spp. isolates and did not appear to contribute to cephamycin resistance. Two strains of B. fragilis contained the metallo-β-lactamase gene cfiA, but they did not produce the protein product. Gene tetQ was detected in about 81, 44, and 63 % of B. fragilis isolates, other Bacteroides spp., and Prevotella spp. isolates, respectively. The ermF gene was detected in 25, 13, 56, 64, and 16 % of Bacteroides spp., Prevotella spp., Fusobacterium spp., B. wadsworthia, and anaerobic cocci, respectively. Gene mefA was found in only 10 % of the B. fragilis strains and 3 % of the non-B. fragilis strains. Genes nim and ermB were not detected in any isolate. Substitution at position 82 (Ser to Phe) in gyrA was detected in B. fragilis isolates that were less susceptible or resistant to moxifloxacin. This study is the first report on the distribution of resistance genes in anaerobes isolated from intra-abdominal infections in Japan. We expect that the results might help in understanding the resistance mechanisms of specific anaerobes.

  2. Quantitative Microbiological Study of Human Carious Dentine by Culture and Real-Time PCR: Association of Anaerobes with Histopathological Changes in Chronic Pulpitis

    Science.gov (United States)

    Martin, F. Elizabeth; Nadkarni, Mangala A.; Jacques, Nicholas A.; Hunter, Neil

    2002-01-01

    The bacteria found in carious dentine were correlated with the tissue response of the dental pulps of 65 teeth extracted from patients with advanced caries and pulpitis. Standardized homogenates of carious dentine were plated onto selective and nonselective media under anaerobic and microaerophilic conditions. In addition, real-time PCR was used to quantify the recovery of anaerobic bacteria. Primers and fluorogenic probes were designed to detect the total anaerobic microbial load, the genera Prevotella and Fusobacterium, and the species Prevotella melaninogenica, Porphyromonas endodontalis, Porphyromonas gingivalis, and Micromonas (formerly Peptostreptococcus) micros. The pulpal pathology was categorized according to the cellular response and degenerative changes. Analysis of cultured bacteria showed a predominance of gram-positive microorganisms, particularly lactobacilli. Gram-negative bacteria were also present in significant numbers with Prevotella spp., the most numerous anaerobic group cultured. Real-time PCR analysis indicated a greater microbial load than that determined by colony counting. The total number of anaerobes detected was 41-fold greater by real-time PCR than by colony counting, while the numbers of Prevotella and Fusobacterium spp. detected were 82- and 2.4-fold greater by real-time PCR than by colony counting, respectively. Real-time PCR also identified M. micros, P. endodontalis, and P. gingivalis in 71, 60, and 52% of carious samples, respectively. Correlation matrices of the real-time PCR data revealed significant positive associations between M. micros and P. endodontalis detection and inflammatory degeneration of pulpal tissues. These anaerobes have been strongly implicated in endodontic infections that occur as sequelae to carious pulpitis. Accordingly, the data suggest that the presence of high levels of these bacteria in carious lesions may be indicative of irreversible pulpal pathology. PMID:11980945

  3. Microbial Signature Profiles of Periodontally Healthy and Diseased Patients

    Science.gov (United States)

    Lourenço, Talita Gomes Baêta; Heller, Débora; da Silva-Boghossian, Carina Maciel; Cotton, Sean L.; Paster, Bruce J.; Colombo, Ana Paula Vieira

    2014-01-01

    Aim To determine microbial profiles that discriminate periodontal health from different forms of periodontal diseases. Methods Subgingival biofilm was obtained from patients with periodontal health (27), gingivitis (11), chronic periodontitis (35) and aggressive periodontitis (24), and analyzed for the presence of >250 species/phylotypes using HOMIM. Microbial differences among groups were examined by Mann-Whitney. Regression analyses were performed to determine microbial risk indicators of disease. Results Putative and potential new periodontal pathogens were more prevalent in subjects with periodontal diseases than periodontal health. Detection of Porphyromonas endodontalis/Porphyromonas spp. (OR 9.5 [1.2–73.1]) and Tannerella forsythia (OR 38.2 [3.2–450.6]), and absence of Neisseria polysaccharea (OR 0.004 [0–0.15]) and Prevotella denticola (OR 0.014 [0–0.49], pdisease. Presence of Aggregatibacter actinomycetemcomitans (OR 29.4 [3.4–176.5]), Cardiobacterium hominis (OR 14.9 [2.3–98.7]), Peptostreptococcaceae sp. (OR 35.9 [2.7–483.9]), P. alactolyticus (OR 31.3 [2.1–477.2]), and absence of Fretibacterium spp. (OR 0.024 [0.002–0.357]), Fusobacterium naviforme/Fusobacterium nucleatum ss vincentii (OR 0.015 [0.001–0.223]), Granulicatella adiacens/Granulicatella elegans (OR 0.013 [0.001–0.233], pdiseases. Such profiles may be used to establish risk of disease. PMID:25139407

  4. Prevalent bacterial species and novel phylotypes in advanced noma lesions.

    Science.gov (United States)

    Paster, B J; Falkler Jr, W A; Enwonwu, C O; Idigbe, E O; Savage, K O; Levanos, V A; Tamer, M A; Ericson, R L; Lau, C N; Dewhirst, F E

    2002-06-01

    The purpose of this study was to determine the bacterial diversity in advanced noma lesions using culture-independent molecular methods. 16S ribosomal DNA bacterial genes from DNA isolated from advanced noma lesions of four Nigerian children were PCR amplified with universally conserved primers and spirochetal selective primers and cloned into Escherichia coli. Partial 16S rRNA sequences of approximately 500 bases from 212 cloned inserts were used initially to determine species identity or closest relatives by comparison with sequences of known species or phylotypes. Nearly complete sequences of approximately 1,500 bases were obtained for most of the potentially novel species. A total of 67 bacterial species or phylotypes were detected, 25 of which have not yet been grown in vitro. Nineteen of the species or phylotypes, including Propionibacterium acnes, Staphylococcus spp., and the opportunistic pathogens Stenotrophomonas maltophilia and Ochrobactrum anthropi were detected in more than one subject. Other known species that were detected included Achromobacter spp., Afipia spp., Brevundimonas diminuta, Capnocytophaga spp., Cardiobacterium sp., Eikenella corrodens, Fusobacterium spp., Gemella haemoylsans, and Neisseria spp. Phylotypes that were unique to noma infections included those in the genera Eubacterium, Flavobacterium, Kocuria, Microbacterium, and Porphyromonas and the related Streptococcus salivarius and genera Sphingomonas and TREPONEMA: Since advanced noma lesions are infections open to the environment, it was not surprising to detect species not commonly associated with the oral cavity, e.g., from soil. Several species previously implicated as putative pathogens of noma, such as spirochetes and Fusobacterium spp., were detected in at least one subject. However, due to the limited number of available noma subjects, it was not possible at this time to associate specific species with the disease.

  5. Oral associated bacterial infection in horses: studies on the normal anaerobic flora from the pharyngeal tonsillar surface and its association with lower respiratory tract and paraoral infections.

    Science.gov (United States)

    Bailey, G D; Love, D N

    1991-02-15

    Two hundred and seventy bacterial isolates were obtained from the pharyngeal tonsillar surface of 12 normal horses and 98 obligatory anaerobic bacteria were characterised. Of these, 57 isolates belonging to 7 genera (Peptostreptococcus (1); Eubacterium (9); Clostridium (6); Veillonella (6); Megasphera (1); Bacteroides (28); Fusobacterium (6)) were identified, and 16 of these were identified to species level (P. anaerobius (1); E. fossor (9); C. villosum (1); B. fragilis (1); B. tectum (2); B. heparinolyticus (2)). Three hundred and twenty isolates were obtained from 23 samples from horses with lower respiratory tract (LRT) or paraoral (PO) bacterial infections. Of the 143 bacteria selected for detailed characterisation, obligate anaerobes accounted for 100 isolates, facultative anaerobes for 42 isolates and obligate aerobes for one isolate. Phenotypic characterisation separated 99 of the isolates into 14 genera. Among the obligately anaerobic species, Gram-positive cocci including P. anaerobius comprised 25% of isolates, E. fossor 11% and other Gram-positive rods (excluding Clostridium sp.) 18% of isolates. The Gram-negative rods comprised B. fragilis 5%, B. heparinolyticus 5%, asaccharolytic pigmented Bacteroides 3% and other Bacteroides 13%, while a so-far unnamed species of Fusobacterium (7%), and Gram-negative corroding rods (3%) were isolated. Among the facultatively anaerobic isolates, S. equi subsp. zooepidemicus accounted for 31% of isolates, followed by Pasteurella spp. 19%, Escherichia coli 17%, Actinomyces spp. 9%, Streptococcus spp. 9%. Incidental facultative isolates were Enterococcus spp. 2%, Enterobacter cloaceae 2%, Actinobacillus spp. 2% and Gram-negative corroding rods 5%. On the basis of the similarities (as determined by DNA hybridization data and/or phenotypic characteristics) of some of the bacterial species (e.g. E. fossor and B. heparinolyticus) isolated from both the normal pharyngeal tonsillar surfaces and LRT and PO diseases of horses, it

  6. Bacterial infection of the lower respiratory tract in 34 horses.

    Science.gov (United States)

    Racklyeft, D J; Love, D N

    2000-08-01

    To investigate associations between the bacteriology and aspects of history, clinical presentation, outcome and pathology of lower respiratory tract disease of 34 horses. Detailed aerobic and anaerobic bacteriological investigations were performed on clinical specimens from horses with pneumonia, lung abscessation and necrotic pneumonia with or without pleurisy in an attempt to identify those bacteria that might contribute to the initiation and progression of infection. Bacteria were cultured from 33 of the 34 horses. In ten cases, only aerobic/facultatively anaerobic isolates were cultured while aerobic/facultatively anaerobic bacteria and obligately anaerobic bacteria were isolated in the other 23 cases. Moderate to large numbers of anaerobic bacteria were isolated only when the estimated duration of illness was at least five days. Bacteria were not cultured from 12 of the pleural fluid samples but were always cultured from pulmonary samples (either transtracheal aspirates from live horses or pulmonary lesions at necropsy). Streptococcus equi subsp zooepidemicus was isolated in the three cases where only one bacterial species was cultured. In the other 30 cases, multiple species were isolated. These included most often and in greatest numbers, Streptococcus equi subsp zooepidemicus, Pasteurellaceae, Escherichia coli, anaerobic cocci, Eubacterium fossor, Bacteroides tectum, Prevotella heparinolytica, Fusobacterium spp, and pigmented members of the genera Prevotella and Porphyromonas. Aerobic/facultatively anaerobic organisms were isolated from 97% of horses, while obligately anaerobic organisms were cultured from 68% of horses. There was no association between the isolation of any specific bacterium and the outcome of disease. However, obligately anaerobic bacteria (such as anaerobic cocci, Bacteroides tectum, P heparinolytica and Fusobacterium spp) and the facultatively anaerobic species Escherichia coli, were recovered more commonly from horses that died or were

  7. Afecções podais em vacas da bacia leiteira de Rondon do Pará Foot disorders in cows from basin milk of Rondon do Pará

    Directory of Open Access Journals (Sweden)

    José Alcides S. Silveira

    2009-11-01

    Full Text Available As doenças digitais estão entre as principais causas de performance reduzida de rebanhos leiteiros. Com o objetivo de determinar a prevalência, classificar clinicamente e estabelecer os fatores epidemiológicos das enfermidades podais em vacas da bacia leiteira de Rondon do Pará, foram avaliadas 1.236 vacas, das quais 275 apresentaram pelo menos um tipo de lesão podal. Identificou-se 655 lesões, o que resultou em uma prevalência de 22,25%. As enfermidades mais frequentes foram hiperplasia interdigital (80,92%, necrobacilose interdigital (6,11% e cascos com crescimento excessivo (6,42%. Os membros pélvicos foram os mais acometidos (61,83% e o espaço interdigital, tanto nos membros torácicos (36,34%, quanto nos pélvicos (48,09%, a região digital acometida com maior frequência. O estudo epidemiológico mostrou que características ambientais tais como relevo montanhoso, pastagem em formação com presença de troncos e galhos de árvores, irregularidades nos pisos dos currais, presença de piçarra e lama podem favorecer o aparecimento das lesões podais. Constatou-se a ausência de medidas de controle e profilaxia de afecções que acometem os cascos em 95,5% das propriedades estudadas. O exame clínico específico do casco foi eficiente no diagnóstico das enfermidades.Foot diseases are some of the main causes of low performance in dairy cows. In order to determine the prevalence, clinical classification and to establish the epidemio-logical factors of foot diseases in cows from basin milk of Rondon do Pará, 1,236 cows were evaluated and 275 had at least one type of foot injury. A total of 655 lesions was showed which a prevalence of 22.25%. Most frequent lesions were interdigital hyperplasia (80.92%, claw overgrowth (6.42%, and interdigital necrobacillosis (6.11%. Hind limbs were most frequently affected (61.83%, and lesions affecting the interdigital space were the most common in both the thoracic (36.34% and hind limbs (48

  8. Clinical characteristics and antimicrobial susceptibilities of anaerobic bacteremia in an acute care hospital.

    Science.gov (United States)

    Tan, Thean Yen; Ng, Lily Siew Yong; Kwang, Lee Ling; Rao, Suma; Eng, Li Ching

    2017-02-01

    This study investigated the clinical features of anaerobic bacteraemia in an acute-care hospital, and evaluated the antimicrobial susceptibility of these isolates to commonly available antibiotics. Microbiological and epidemiological data from 2009 to 2011were extracted from the laboratory information system and electronic medical records. One hundred and eleven unique patient episodes consisting of 116 anaerobic isolates were selected for clinical review and antibiotic susceptibility testing. Susceptibilities to amoxicillin-clavulanate, clindamycin, imipenem, metronidazole, moxifloxacin, penicillin and piperacillin-tazobactam were performed using Etest strips with categorical interpretations according to current CLSI breakpoints. Metronidazole-resistant and carbapenem-resistant anaerobic Gram-negative bacilli were screened for the nim and cfiA genes. Clinical data was obtained retrospectively from electronic medical records. During the 3 year period, Bacteroides fragilis group (41%), Clostridium species (14%), Propionibacterium species (9%) and Fusobacterium species (6%) were the most commonly isolated anaerobes. Patients with anaerobic bacteraemia that were included in the study were predominantly above 60 years of age, with community-acquired infections. The most commonly used empiric antibiotic therapies were beta-lactam/beta-lactamase inhibitor combinations (44%) and metronidazole (10%). The crude mortality was 25%, and appropriate initial antibiotic therapy was not significantly associated with improved survival. Intra-abdominal infections (39%) and soft-tissue infections (33%) accounted for nearly three-quarters of all bacteraemia. Antibiotics with the best anaerobic activity were imipenem, piperacillin-tazobactam, amoxicillin-clavulanate and metronidazole, with in-vitro susceptibility rates of 95%, 95%, 94% and 92% respectively. Susceptibilities to penicillin (31%), clindamycin (60%) and moxifloxacin (84%) were more variable. Two multidrug

  9. Detection of putative periodontal pathogens in subgingival specimens of dogs Detecção de patógenos periodontais em amostras subgengivais de cães

    Directory of Open Access Journals (Sweden)

    Sheila Alexandra Belini Nishiyama

    2007-03-01

    Full Text Available In this study, the presence of putative periodontal organisms, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, Fusobacterium nucleatum,Dialister pneumosintes,Actinobacillus actinomycetemcomitans,Campylobacter rectus,Eikenella corrodens and Treponema denticola were examined from subgingival samples of 40 dogs of different breeds with (25 and without (15 periodontitis, by using the PCR method. The PCR products of each species showed specific amplicons. Of the 25 dogs with periodontitis, P. gingivalis was detected in 16 (64% samples, C. rectus in 9 (36%, A. actinomycetemcomitans in 6 (24%, P. intermedia in 5 (20%, T. forsythensis in 5 (20%, F. nucleatum in 4 (16% and E. corrodens in 3 (12%. T. denticola and D. pneumosintes were not detected in clinical samples from dogs with periodontitis. Moreover, P. gingivalis was detected only in one (6.66% crossbred dog without periodontitis. Our results show that these microorganisms are present in periodontal microbiota of dogs with periodontitits, and it is important to evaluate the role of these putative periodontal microorganisms play in the periodontitis in household pets particularly, dogs in ecologic and therapeutic terms, since these animals might acquire these periodontopahogens from their respective owners.Neste estudo, a presença de patógenos periodontais, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, Fusobacterium nucleatum, Dialister pneumosintes, Actinobacillus actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens e Treponema denticola foi determinada por PCR, em amostras subgengivais de 40 cães com (25 e sem (15 doença periodontal. Os produtos amplificados pelo PCR para cada espécie bacteriana mostraram amplicons específicos. Dos 25 cães apresentando doença periodontal, P. gingivalis foi detectado em 16 amostras (64%, C. rectus em 9 (36%, A. actinomycetemcomitans em 6 (24%, P. intermedia em 5 (20%, T. forsythensis em 5 (20

  10. Comparing the Healthy Nose and Nasopharynx Microbiota Reveals Continuity As Well As Niche-Specificity

    Directory of Open Access Journals (Sweden)

    Ilke De Boeck

    2017-11-01

    Full Text Available To improve our understanding of upper respiratory tract (URT diseases and the underlying microbial pathogenesis, a better characterization of the healthy URT microbiome is crucial. In this first large-scale study, we obtained more insight in the URT microbiome of healthy adults. Hereto, we collected paired nasal and nasopharyngeal swabs from 100 healthy participants in a citizen-science project. High-throughput 16S rRNA gene V4 amplicon sequencing was performed and samples were processed using the Divisive Amplicon Denoising Algorithm 2 (DADA2 algorithm. This allowed us to identify the bacterial richness and diversity of the samples in terms of amplicon sequence variants (ASVs, with special attention to intragenus variation. We found both niches to have a low overall species richness and uneven distribution. Moreover, based on hierarchical clustering, nasopharyngeal samples could be grouped into some bacterial community types at genus level, of which four were supported to some extent by prediction strength evaluation: one intermixed type with a higher bacterial diversity where Staphylococcus, Corynebacterium, and Dolosigranulum appeared main bacterial members in different relative abundances, and three types dominated by either Moraxella, Streptococcus, or Fusobacterium. Some of these bacterial community types such as Streptococcus and Fusobacterium were nasopharynx-specific and never occurred in the nose. No clear association between the nasopharyngeal bacterial profiles at genus level and the variables age, gender, blood type, season of sampling, or common respiratory allergies was found in this study population, except for smoking showing a positive association with Corynebacterium and Staphylococcus. Based on the fine-scale resolution of the ASVs, both known commensal and potential pathogenic bacteria were found within several genera – particularly in Streptococcus and Moraxella – in our healthy study population. Of interest, the

  11. Isolamento e identificação da microbiota periodontal de cães da raça Pastor Alemão Isolate and identify of periodontal microbiota of German Shepherd dogs

    Directory of Open Access Journals (Sweden)

    Carla Afonso da Silva Bitencourt Braga

    2005-04-01

    Full Text Available A microbiota indígena gengival de cães não está totalmente descrita, sendo sua identificação uma etapa importante no estabelecimento da etiopatogenia e terapia da doença periodontal. O objetivo deste trabalho foi isolar e identificar a microbiota periodontal de cães da raça Pastor Alemão, considerando sítios saudáveis e com doença periodontal. Foram utilizados 29 cães, com idade variando de três a seis anos, sendo analisados espécimes clínicos de sítios periodontais saudáveis de 12 cães e sítios com periodontite de outros 17. Foram isoladas 672 amostras microbianas, com predomínio dos gêneros Pasteurella, Staphylococcus, Porphyromonas e Fusobacterium. A microbiota dos sítios saudáveis equiparou-se à dos sítios doentes, tratando-se de uma microbiota indígena. A microbiota dos sítios doentes apresentou-se aumentada em relação a dos sítios saudáveis, indicando mudança do ambiente do sítio periodontal.The indigenous gingival microbiota of dogs is not totally described, although such identification is an important step to establish the etiopathogenesis and adequate therapy for the periodontal disease. The aims of this study were to isolate and identify the periodontal microbiota of German Shepherd dogs from healthy and with periodontal desease sites. Twenty nine German Shepherd dogs from three to six years of age were used in this study. Clinical specimens were analysed from healthy periodontal sites of 12 dogs and sites presenting gingivitis of 17 dogs. A total amount of 672 microbial samples, were isolated where the predominant genera were Pasteurella spp., Staphylococcus spp., Porphyromonas spp. and Fusobacterium spp. The microbiological population of the affected sites was similar to the healthy sites, consisting on an indigenous microbiota. The microbiota on the affecteded sites was higher in number than on the healthy sites, showing change in the environment of the periodontal sites.

  12. Robust species taxonomy assignment algorithm for 16S rRNA NGS reads: application to oral carcinoma samples

    Directory of Open Access Journals (Sweden)

    Nezar Noor Al-Hebshi

    2015-09-01

    Full Text Available Background: Usefulness of next-generation sequencing (NGS in assessing bacteria associated with oral squamous cell carcinoma (OSCC has been undermined by inability to classify reads to the species level. Objective: The purpose of this study was to develop a robust algorithm for species-level classification of NGS reads from oral samples and to pilot test it for profiling bacteria within OSCC tissues. Methods: Bacterial 16S V1-V3 libraries were prepared from three OSCC DNA samples and sequenced using 454's FLX chemistry. High-quality, well-aligned, and non-chimeric reads ≥350 bp were classified using a novel, multi-stage algorithm that involves matching reads to reference sequences in revised versions of the Human Oral Microbiome Database (HOMD, HOMD extended (HOMDEXT, and Greengene Gold (GGG at alignment coverage and percentage identity ≥98%, followed by assignment to species level based on top hit reference sequences. Priority was given to hits in HOMD, then HOMDEXT and finally GGG. Unmatched reads were subject to operational taxonomic unit analysis. Results: Nearly, 92.8% of the reads were matched to updated-HOMD 13.2, 1.83% to trusted-HOMDEXT, and 1.36% to modified-GGG. Of all matched reads, 99.6% were classified to species level. A total of 228 species-level taxa were identified, representing 11 phyla; the most abundant were Proteobacteria, Bacteroidetes, Firmicutes, Fusobacteria, and Actinobacteria. Thirty-five species-level taxa were detected in all samples. On average, Prevotella oris, Neisseria flava, Neisseria flavescens/subflava, Fusobacterium nucleatum ss polymorphum, Aggregatibacter segnis, Streptococcus mitis, and Fusobacterium periodontium were the most abundant. Bacteroides fragilis, a species rarely isolated from the oral cavity, was detected in two samples. Conclusion: This multi-stage algorithm maximizes the fraction of reads classified to the species level while ensuring reliable classification by giving priority to the

  13. Illumina MiSeq 16S amplicon sequence analysis of bovine respiratory disease associated bacteria in lung and mediastinal lymph node tissue.

    Science.gov (United States)

    Johnston, Dayle; Earley, Bernadette; Cormican, Paul; Murray, Gerard; Kenny, David Anthony; Waters, Sinead Mary; McGee, Mark; Kelly, Alan Kieran; McCabe, Matthew Sean

    2017-05-02

    Bovine respiratory disease (BRD) is caused by growth of single or multiple species of pathogenic bacteria in lung tissue following stress and/or viral infection. Next generation sequencing of 16S ribosomal RNA gene PCR amplicons (NGS 16S amplicon analysis) is a powerful culture-independent open reference method that has recently been used to increase understanding of BRD-associated bacteria in the upper respiratory tract of BRD cattle. However, it has not yet been used to examine the microbiome of the bovine lower respiratory tract. The objective of this study was to use NGS 16S amplicon analysis to identify bacteria in post-mortem lung and lymph node tissue samples harvested from fatal BRD cases and clinically healthy animals. Cranial lobe and corresponding mediastinal lymph node post-mortem tissue samples were collected from calves diagnosed as BRD cases by veterinary laboratory pathologists and from clinically healthy calves. NGS 16S amplicon libraries, targeting the V3-V4 region of the bacterial 16S rRNA gene were prepared and sequenced on an Illumina MiSeq. Quantitative insights into microbial ecology (QIIME) was used to determine operational taxonomic units (OTUs) which corresponded to the 16S rRNA gene sequences. Leptotrichiaceae, Mycoplasma, Pasteurellaceae, and Fusobacterium were the most abundant OTUs identified in the lungs and lymph nodes of the calves which died from BRD. Leptotrichiaceae, Fusobacterium, Mycoplasma, Trueperella and Bacteroides had greater relative abundances in post-mortem lung samples collected from fatal cases of BRD in dairy calves, compared with clinically healthy calves without lung lesions. Leptotrichiaceae, Mycoplasma and Pasteurellaceae showed higher relative abundances in post-mortem lymph node samples collected from fatal cases of BRD in dairy calves, compared with clinically healthy calves without lung lesions. Two Leptotrichiaceae sequence contigs were subsequently assembled from bacterial DNA-enriched shotgun sequences

  14. Aerobic and anaerobic bacteria in tonsils of children with recurrent tonsillitis.

    Science.gov (United States)

    Brook, I; Yocum, P; Friedman, E M

    1981-01-01

    Tonsils were obtained from 50 children suffering from recurrent tonsillitis. Patients' ages ranged from 2.5 to 17 years (mean 6 years); 29 were males and 21 females. The tonsils were sectioned in half after heat searing of the surface and the core material was cultured for aerobic and anaerobic microorganisms. Mixed aerobic and anaerobic flora was obtained in all patients, yielding an average of 7.8 isolates (4.1 anaerobes and 3.7 aerobes) per specimen. There were 207 anaerobes isolated. The predominant isolates were 101 Bacteroides sp (including 10 B fragilis group, and 47 B melaninogenicus group), 29 Fusobacterium sp, 34 Gram-positive anaerobic cocci (25 Peptococcus sp and 9 Peptostreptococcus sp) and 16 Veillonella sp. There were 185 aerobic isolates. The predominant isolates were 41 alpha-hemolytic streptococci, 24 Staphylococcus aureus, 19 beta-hemolytic streptococci (11 group A, 4 group B, and 2 each group C and F), 14 Haemophilus sp (including 12 H influenzae type B) and 5 H parainfluenzae. Beta-lactamase production was noted in 56 isolates recovered from 37 tonsils. These were all isolates of S aureus (24) and B fragilis (10), 15 of 47 B melaninogenicus (32%), 5 of the 12 B oralis (42%), and 2 of 12 H influenzae type B (17%). Our findings indicate the polymicrobial aerobic and anaerobic nature of deep tonsillar flora in children with recurrent tonsillitis, and demonstrate the presence of many beta-lactamase-producing organisms in 74% of the patients.

  15. Immune status, antibiotic medication and pH are associated with changes in the stomach fluid microbiota.

    Science.gov (United States)

    von Rosenvinge, Erik C; Song, Yang; White, James R; Maddox, Cynthia; Blanchard, Thomas; Fricke, W Florian

    2013-07-01

    The stomach acts as a barrier to ingested microbes, thereby influencing the microbial ecology of the entire gastrointestinal (GI) tract. The stomach microbiota and the role of human host and environmental factors, such as health status or medications, in shaping its composition remain largely unknown. We sought to characterize the bacterial and fungal microbiota in the stomach fluid in order to gain insights into the role of the stomach in GI homeostasis. Gastric fluid was collected from 25 patients undergoing clinically indicated upper endoscopy. DNA isolates were used for PCR amplification of bacterial 16S ribosomal RNA (rRNA) genes and fungal internal transcribed spacers (ITS). RNA isolates were used for 16S rRNA cDNA generation and subsequent PCR amplification. While all stomach fluid samples are dominated by the phyla Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria and Fusobacteria (>99% of sequence reads), the transcriptionally active microbiota shows significant reduction in Actinobacteria (34%) and increase in Campylobacter (444%) (PpH >4 (70%; P<0.05). Immunosuppression correlates with decreased abundance of Prevotella (24%), Fusobacterium (2%) and Leptotrichia (6%) and increased abundance of Lactobacillus (3844%) (P<0.003). We have generated the first in-depth characterization of the human gastric fluid microbiota, using bacterial 16S rRNA gene and transcript, and fungal ITS amplicon sequencing and provide evidence for a significant impact of the host immune status on its composition with likely consequences for human health.

  16. Health risk to residents and stimulation to inherent bacteria of various heavy metals in soil.

    Science.gov (United States)

    Zhang, Juan; Wang, Li-Hong; Yang, Jun-Cheng; Liu, Hui; Dai, Jiu-Lan

    2015-03-01

    The toxicities and effects of various metals and metalloids would be misunderstood by health risks based on their concentrations, when their effects on bacterial and ecological functions in soil are disregarded. This study investigated the concentrations and health risks of heavy metals, soil properties, and bacterial 16S rRNA gene in soil around the largest fresh water lake in North China. The health risks posed by Mn and As were higher than those of other heavy metals and metalloids. Mn, As, and C were significantly correlated with the bacterial species richness indices. According to canonical correspondence analysis, species richness was mainly affected by Mn, Pb, As, and organic matter, while species evenness was mainly affected by Mn, pH, N, C, Cd, and Pb. Covariable analysis confirmed that most effects of metals on bacterial diversity were attributed to the combined effects of metals and soil properties rather than single metals. Most bacteria detected in (almost) all soil were identified as Gammaproteobacteria. Specific bacteria belonging to Proteobacteria (Gamma, Alpha, Epsilon, and Beta), Firmicutes, Actinobacteria, Cyanobacterium, Nitrospirae, and Fusobacterium were only identified in soil with high concentrations of Mn, Pb, and As, indicating their remediation potency. Bacterial abilities and mechanisms in pollutant resistance and element cycling in the region were also discussed. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. The protective effect of recombinant FomA-expressing Lactobacillus acidophilus against periodontal infection.

    Science.gov (United States)

    Ma, Li; Ding, Qinfeng; Feng, Xiping; Li, Fei

    2013-10-01

    A number of studies have shown that the outer membrane protein FomA found in Fusobacterium nucleatum demonstrates great potential as an immune target for combating periodontitis. Lactobacillus acidophilus is a useful antigen delivery vehicle for mucosal immunisation, and previous studies by our group have shown that L. acidophilus acts as a protective factor in periodontal health. In this study, making use of the immunogenicity of FomA and the probiotic properties of L. acidophilus, we constructed a recombinant form of L. acidophilus expressing the FomA protein and detected the FomA-specific IgG in the serum and sIgA in the saliva of mice through oral administration with the recombinant strains. When serum containing FomA-specific antibodies was incubated with the F. nucleatum in vitro, the number of Porphyromonas gingivalis cells that coaggregated with the F. nucleatum cells was significantly reduced. Furthermore, a mouse gum abscess model was successfully generated, and the range of gingival abscesses in the immune mice was relatively limited compared with the control group. The level of IL-1β in the serum and local gum tissues of the immune mice was consistently lower than in the control group. Our findings indicated that oral administration of the recombinant L. acidophilus reduced the risk of periodontal infection with P. gingivalis and F. nucleatum.

  18. Lactobacillus salivarius WB21--containing tablets for the treatment of oral malodor: a double-blind, randomized, placebo-controlled crossover trial.

    Science.gov (United States)

    Suzuki, Nao; Yoneda, Masahiro; Tanabe, Kazunari; Fujimoto, Akie; Iha, Kosaku; Seno, Kei; Yamada, Kazuhiko; Iwamoto, Tomoyuki; Masuo, Yosuke; Hirofuji, Takao

    2014-04-01

    This study evaluated the effect of probiotic intervention using lactobacilli on oral malodor. We conducted a 14-day, double-blind, placebo-controlled, randomized crossover trial of tablets containing Lactobacillus salivarius WB21 (2.0 × 10(9) colony-forming units per day) or placebo taken orally by patients with oral malodor. Organoleptic test scores significantly decreased in both the probiotic and placebo periods compared with the respective baseline scores (P < .001 and P = .002), and no difference was detected between periods. In contrast, the concentration of volatile sulfur compounds (VSCs) (P = .019) and the average probing pocket depth (P = .001) decreased significantly in the probiotic period compared with the placebo period. Bacterial quantitative analysis found significantly lower levels of ubiquitous bacteria (P = .003) and Fusobacterium nucleatum (P = .020) in the probiotic period. These results indicated that daily oral consumption of tablets containing probiotic lactobacilli could help to control oral malodor and malodor-related factors. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. Effects of oil drops containing Lactobacillus salivarius WB21 on periodontal health and oral microbiota producing volatile sulfur compounds.

    Science.gov (United States)

    Suzuki, Nao; Tanabe, Kazunari; Takeshita, Toru; Yoneda, Masahiro; Iwamoto, Tomoyuki; Oshiro, Sueko; Yamashita, Yoshihisa; Hirofuji, Takao

    2012-03-01

    The objective of this paper is to evaluate the effects of oil drops containing Lactobacillus salivarius WB21 on periodontal health and oral microbiota producing volatile sulfur compounds (VSCs). For this study, 42 subjects were randomly assigned to receive oil samples containing L. salivarius WB21 or a placebo for two weeks. Oral assessment and saliva collection were performed on days 1 and 15. Bacterial analysis was performed using the real-time polymerase chain reaction and terminal restriction fragment length polymorphism (T-RFLP). In both the experimental and placebo groups, the average probing depth, number of periodontal pockets, and the percentage of bleeding on probing (BOP) decreased while stimulated salivary flow increased on day 15. BOP was reduced in the experimental group compared with the placebo group (P = 0.010). In the experimental group, total bacterial numbers decreased, and the number of L. salivarius increased. The number of Prevotella intermedia, which is correlated with hydrogen sulfide concentration in mouth air, increased in the placebo group and did not change in the experimental group. T-RFLP analysis found that the peak area proportions representing Porphyromonas gingivalis, P. intermedia, Tannerella forsythensis, and Fusobacterium nucleatum decreased in the experimental group, although there was no significant change in the bacterial composition. Thus we observed oil drops containing L. salivarius WB21 improved BOP and inhibited the reproduction of total and VSC-producing periodontopathic bacteria compared with the placebo group, but also showed the limit of its efficacy in controlling VSCs producing and periodontal pathogens.

  20. Epithelial cell pro-inflammatory cytokine response differs across dental plaque bacterial species.

    Science.gov (United States)

    Stathopoulou, Panagiota G; Benakanakere, Manjunatha R; Galicia, Johnah C; Kinane, Denis F

    2010-01-01

    The dental plaque is comprised of numerous bacterial species, which may or may not be pathogenic. Human gingival epithelial cells (HGECs) respond to perturbation by various bacteria of the dental plaque by production of different levels of inflammatory cytokines, which is a putative reflection of their virulence. The aim of the current study was to determine responses in terms of interleukin (IL)-1beta, IL-6, IL-8 and IL-10 secretion induced by Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and Streptococcus gordonii in order to gauge their virulence potential. HGECs were challenged with the four bacterial species, live or heat killed, at various multiplicity of infections and the elicited IL-1beta, IL-6, IL-8 and IL-10 responses were assayed by enzyme-linked immunosorbent assay. Primary HGECs challenged with live P. gingivalis produced high levels of IL-1beta, while challenge with live A. actinomycetemcomitans gave high levels of IL-8. The opportunistic pathogen F. nucleatum induces the highest levels of pro-inflammatory cytokines, while the commensal S. gordonii is the least stimulatory. We conclude that various dental plaque biofilm bacteria induce different cytokine response profiles in primary HGECs that may reflect their individual virulence or commensal status.

  1. Longitudinal study on clinical and microbial analysis of periodontal status in pregnancy

    Directory of Open Access Journals (Sweden)

    Fernanda Campos MACHADO

    Full Text Available Abstract This study was aimed to provide a longitudinal overview of the subgingival bacterial microbiome using fluorescence in situ hybridization (FISH technique, in women in the second trimester of pregnancy (between 14 and 24 weeks, and 48 h and 8 weeks postpartum. Of 31 women evaluated during pregnancy, 24 returned for the 48-h and 18 for their 8-week exams postpartum. Probing depth (PD, bleeding on probing, clinical attachment level, and presence of calculus were recorded. Subgingival plaque samples were collected, and FISH was used to identify the numbers of eight periodontal pathogens. Friedman test was used to compare differences between follow-up examinations, followed by a multiple comparison test for a post hoc pairwise comparison. Clinically, a significantly greater number of teeth with PD = 4–5 mm were found during pregnancy than on postpartum examinations. Microbial analysis showed a statistically significant decrease in cell count over the study period for Prevotella nigrescens. P. intermedia, Campylobacter rectus, and Porphyromonas gingivalis also decrease, although not significantly, and Aggregatibacter actinomycetemcomitans increased. No significant changes were found for Fusobacterium nucleatum, Treponema denticola, or Tannerella forsythia. Our data demonstrate a change in the subgingival microbiota during pregnancy, at least for P. nigrescens.

  2. Effect of Lactobacillus rhamnosus and Bifidobacterium lactis on gingival health, dental plaque, and periodontopathogens in adolescents: a randomised placebo-controlled clinical trial.

    Science.gov (United States)

    Alanzi, A; Honkala, S; Honkala, E; Varghese, A; Tolvanen, M; Söderling, E

    2018-04-10

    To determine the effect of a probiotic combination of Lactobacillus rhamnosus GG (LGG) and Bifidobacterium lactis BB-12 on the gingival health, dental plaque accumulation, and the oral carriage of four putative periodontal pathogens in healthy adolescents. 108 schoolboys, aged 13-15 years, participated in this study. They were divided into two groups: probiotics (n=54) and placebo (n=54). Both groups received two probiotic-laced or placebo lozenges twice a day during a four-week period. Plaque Index (PI) and Gingival Index (GI) were recorded at baseline and after four weeks. Salivary and plaque carriage of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum were also monitored likewise. 101 subjects completed the study. A statistically significant reduction in GI was seen in the probiotic group as compared to the placebo group (P=0.012). A reduction in PI was found for both groups, with no difference observed between the groups after intervention (P=0.819). Probiotic lozenges significantly reduced levels of A. actinomycetemcomitans and F. nucleatum in saliva and plaque (Pplaque (Pbacterial counts of the test group. The short-term daily consumption of LGG and BB-12 probiotic lozenges improved the gingival health in adolescents and decreased the microbial counts of A. actinomycetemcomitans, and P. gingivalis. Hence probiotic supplements may serve as a simple adjunct to standard oral care for promoting the oral health in adolescents.

  3. Changes in microflora in dental plaque from cancer patients undergoing chemotherapy and the relationship of these changes with mucositis: A pilot study.

    Science.gov (United States)

    Vozza, Iole; Caldarazzo, Vito; Ottolenghi, Livia

    2015-05-01

    To assess changes in oral microflora in dental plaque from cancer patients within 7 days of the first course of chemotherapy, and the relationship of the changes with mucositis. Thirty cancer patients, divided into a test group undergoing chemotherapy and a control group no undergoing chemotherapy, were enrolled in this pilot study. Oral microflora were cultured from three samples of dental plaque at t0 (before chemotherapy), t1 (1 day after chemotherapy) and t2 (7 days after chemotherapy). Single and crossed descriptive analyses were used to establish prevalence, and the χ² test was used to establish the statistical significance of the differences observed in distributions (significance level: Pbacterial flora also had periodontal-pathogenic species. No Porphyromonas gingivalis appeared in the test group. Actinobacillus was the least frequently found bacterium among periodontal pathogens in the test group, while Fusobacterium nucleatum was the most frequently found. No significant differences were found in quantitative bacterial changes between t0, t1 and t2 in either the test or control groups, or between the two groups. According to World Health Organization scores, oral mucositis developed in 10 patients (66.6%) in the test group. The results of this pilot study indicate that there were no changes in microflora in dental plaque in cancer patients within 7 days of the first course of chemotherapy. No correlations between oral mucositis and specific microorganisms were assessed.

  4. Community analysis of dental plaque and endotracheal tube biofilms from mechanically ventilated patients.

    Science.gov (United States)

    Marino, Poala J; Wise, Matt P; Smith, Ann; Marchesi, Julian R; Riggio, Marcello P; Lewis, Michael A O; Williams, David W

    2017-06-01

    Mechanically ventilated patients are at risk for developing ventilator-associated pneumonia, and it has been reported that dental plaque provides a reservoir of respiratory pathogens that may aspirate to the lungs and endotracheal tube (ETT) biofilms. For the first time, metataxonomics was used to simultaneously characterize the microbiome of dental plaque, ETTs, and non-directed bronchial lavages (NBLs) in mechanically ventilated patients to determine similarities in respective microbial communities and therefore likely associations. Bacterial 16S rRNA gene sequences from 34 samples of dental plaque, NBLs, and ETTs from 12 adult mechanically ventilated patients were analyzed. No significant differences in the microbial communities of these samples were evident. Detected bacteria were primarily oral species (e.g., Fusobacterium nucleatum, Streptococcus salivarius, Prevotella melaninogenica) with respiratory pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcuspneumoniae, and Haemophilus influenzae) also in high abundance. The high similarity between the microbiomes of dental plaque, NBLs, and ETTs suggests that the oral cavity is indeed an important site involved in microbial aspiration to the lower airway and ETT. As such, maintenance of good oral hygiene is likely to be highly important in limiting aspiration of bacteria in this vulnerable patient group. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Comparative evaluation of subgingival plaque microflora in pregnant and non-pregnant women: A clinical and microbiologic study

    Directory of Open Access Journals (Sweden)

    Rishi Emmatty

    2013-01-01

    Full Text Available Context: Gingival changes in pregnancy have been attributed to changes in the subgingival biofilm related to hormonal variations. Aims: To evaluate the subgingival plaque microflora in pregnant and nonpregnant women to determine if pregnancy induces any alterations in the subgingival plaque and to associate these changes with changes in periodontal status. Settings and Design: Thirty pregnant and 10 nonpregnant women within the age group of 20-35 years having a probing pocket depth (PPD of 3-4 mm were included in the study. The pregnant women were equally divided into 3 groups of 10, each belonging to I, II, and III trimester. Materials and Methods: Plaque index, gingival index, PPD, and microbiologic evaluation for specific bacterial counts for Prevotella intermedia, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Parvimonas micra, and Fusobacterium nucleatum were carried out for all subjects. Statistical Analysis: Mann-Whitney U test. Results: Increase in gingival inflammation was observed in II and III trimester as compared with I trimester and control. Plaque scores did not show any significant difference between pregnant and nonpregnant women. Specific bacterial examination revealed an increase in proportion of P. intermedia in pregnant women of both II and III trimester as compared with I trimester and nonpregnant women. Conclusions: A definite increase in proportions of P. intermedia occurs in subgingival plaque microflora in pregnancy that may be responsible for the exaggerated gingival response.

  6. Comparative study of the effects of photodynamic therapy and conventional therapy on ligature induced peri-implantitis in dogs

    International Nuclear Information System (INIS)

    Hayek, Ricardo Rada Ahmad

    2004-01-01

    Progressive peri-implanter bone losses, which are accompanied by inflammatory process in the soft tissues is referred to as peri-implantitis. The aim of this study was to compare the effects of lethal photosensitisation with the conventional technique on bacterial reduction in ligature induced peri-implantitis in dogs. Seventeen third pre-molars of Labrador dogs were extracted and, immediately after, the implants were submerged. After osteointegration, peri-implantitis was induced. After 4 months, ligature were removed and the same period was waited for natural induction of bacterial plaque. The dogs were randomly divided into two groups. In the conventional group, they were treated with the conventional techniques of mucoperiosteal flaps for scaling the implant surface and irrigate it. In the laser group, only mucoperiosteal scaling was carried out before photodynamic therapy. On the peri-implanter pocket an azulene paste was injected and a GaAlAs low-power laser (λ= 660 nm, P= 30 mW, E= 5,4 J and Δt= 3 min.). Microbiological samples were obtained before and immediately after treatment. One implant was removed to be analyzed by scan electron microscopy to verify contamination on the implant surface. The results of this study showed that Prevotella sp., Fusobacterium e S. Beta-haemolyticus were significantly reduced for the conventional and laser groups. (author)

  7. Pyrosequencing the Canine Faecal Microbiota: Breadth and Depth of Biodiversity

    Science.gov (United States)

    Hand, Daniel; Wallis, Corrin; Colyer, Alison; Penn, Charles W.

    2013-01-01

    Mammalian intestinal microbiota remain poorly understood despite decades of interest and investigation by culture-based and other long-established methodologies. Using high-throughput sequencing technology we now report a detailed analysis of canine faecal microbiota. The study group of animals comprised eleven healthy adult miniature Schnauzer dogs of mixed sex and age, some closely related and all housed in kennel and pen accommodation on the same premises with similar feeding and exercise regimes. DNA was extracted from faecal specimens and subjected to PCR amplification of 16S rDNA, followed by sequencing of the 5′ region that included variable regions V1 and V2. Barcoded amplicons were sequenced by Roche-454 FLX high-throughput pyrosequencing. Sequences were assigned to taxa using the Ribosomal Database Project Bayesian classifier and revealed dominance of Fusobacterium and Bacteroidetes phyla. Differences between animals in the proportions of different taxa, among 10,000 reads per animal, were clear and not supportive of the concept of a “core microbiota”. Despite this variability in prominent genera, littermates were shown to have a more similar faecal microbial composition than unrelated dogs. Diversity of the microbiota was also assessed by assignment of sequence reads into operational taxonomic units (OTUs) at the level of 97% sequence identity. The OTU data were then subjected to rarefaction analysis and determination of Chao1 richness estimates. The data indicated that faecal microbiota comprised possibly as many as 500 to 1500 OTUs. PMID:23382835

  8. Pyrosequencing the canine faecal microbiota: breadth and depth of biodiversity.

    Directory of Open Access Journals (Sweden)

    Daniel Hand

    Full Text Available Mammalian intestinal microbiota remain poorly understood despite decades of interest and investigation by culture-based and other long-established methodologies. Using high-throughput sequencing technology we now report a detailed analysis of canine faecal microbiota. The study group of animals comprised eleven healthy adult miniature Schnauzer dogs of mixed sex and age, some closely related and all housed in kennel and pen accommodation on the same premises with similar feeding and exercise regimes. DNA was extracted from faecal specimens and subjected to PCR amplification of 16S rDNA, followed by sequencing of the 5' region that included variable regions V1 and V2. Barcoded amplicons were sequenced by Roche-454 FLX high-throughput pyrosequencing. Sequences were assigned to taxa using the Ribosomal Database Project Bayesian classifier and revealed dominance of Fusobacterium and Bacteroidetes phyla. Differences between animals in the proportions of different taxa, among 10,000 reads per animal, were clear and not supportive of the concept of a "core microbiota". Despite this variability in prominent genera, littermates were shown to have a more similar faecal microbial composition than unrelated dogs. Diversity of the microbiota was also assessed by assignment of sequence reads into operational taxonomic units (OTUs at the level of 97% sequence identity. The OTU data were then subjected to rarefaction analysis and determination of Chao1 richness estimates. The data indicated that faecal microbiota comprised possibly as many as 500 to 1500 OTUs.

  9. Biofilm forming ability of bacteria isolated from necrotic roots canals of teeth

    Science.gov (United States)

    Alwan, Merriam Ghadhanfar; Usup, Gires; Heng, Lee Yook; Ahmad, Asmat

    2018-04-01

    The growth of microbes in biofilms are associated with repeated and chronic human infections and are extremely resistant to antimicrobial agents. The purpose of this study was to determine the diversity of bacteria from necrotic roots canals of teeth and to detect their biofilm formation ability. A total of 42 bacterial isolates were isolated and identified as belonging to 11 genera. These are Enterococcus sp. (21.4%) followed by Streptococcus sp. (16.8%), Bacillus sp. (11.9%), Peptostreptococcus sp. (9.5%), Staphylococcus sp. (9.5%), Bacteroides sp. (7.1%), Clostridium sp. (7.1%), Actinomyces sp. (7.1%), Fusobacterium sp. (4.76%), Provotella sp. (2.4%) and Chromobacterium sp. (2.4%). Three screening methods for biofilm forming ability were used. Congo Red Agar method (CRA), Tube method (TM) and Microtitre Plate (MTP). From the results, MTP method is a more reliable and quantitative method for the screening and detection of microorganism's ability to form biofilm. This method can be recommended and suggested as a general screening method for the detection of biofilm forming bacteria isolated from roots canals of teeth.

  10. Response to antiseptic agents of periodontal pathogens in in vitro biofilms on titanium and zirconium surfaces.

    Science.gov (United States)

    Sánchez, M C; Fernández, E; Llama-Palacios, A; Figuero, E; Herrera, D; Sanz, M

    2017-04-01

    The aim of this study was to develop in vitro biofilms on SLA titanium (Ti-SLA) and zirconium oxide (ZrO 2 ) surfaces and to evaluate the effect of antiseptic agents on the number of putative periodontal pathogenic species. An in vitro biofilm model was developed on sterile discs of Ti-SLA and ZrO 2 . Three antiseptic agents [chlorhexidine and cetyl-pyridinium-chloride (CHX/CPC), essential oils (EEOOs) and cetyl-peridinium-chloride (CPC)] were applied to 72-h biofilms, immersing discs during 1min in the antiseptic solution, either with or without mechanical disruption. Viable bacteria [colony forming units (CFU/mL)] were measured by quantitative polymerase chain reaction (qPCR) combined with propidium monoazide. A generalized lineal model was constructed to determine the effect of the agents on the viable bacterial counts of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Fusobacterium nucleatum on each surface. The exposure to each antiseptic solution resulted in a statistically significant reductions in the number of viable target species included in the in vitro multi-species biofilm, on both Ti-SLA and ZrO 2 (pzirconium surfaces, in spite of the described structural differences between these bacterial communities. Copyright © 2017 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  11. A dysbiosis index to assess microbial changes in fecal samples of dogs with chronic inflammatory enteropathy.

    Science.gov (United States)

    AlShawaqfeh, M K; Wajid, B; Minamoto, Y; Markel, M; Lidbury, J A; Steiner, J M; Serpedin, E; Suchodolski, J S

    2017-11-01

    Recent studies have identified various bacterial groups that are altered in dogs with chronic inflammatory enteropathies (CE) compared to healthy dogs. The study aim was to use quantitative PCR (qPCR) assays to confirm these findings in a larger number of dogs, and to build a mathematical algorithm to report these microbiota changes as a dysbiosis index (DI). Fecal DNA from 95 healthy dogs and 106 dogs with histologically confirmed CE was analyzed. Samples were grouped into a training set and a validation set. Various mathematical models and combination of qPCR assays were evaluated to find a model with highest discriminatory power. The final qPCR panel consisted of eight bacterial groups: total bacteria, Faecalibacterium, Turicibacter, Escherichia coli, Streptococcus, Blautia, Fusobacterium and Clostridium hiranonis. The qPCR-based DI was built based on the nearest centroid classifier, and reports the degree of dysbiosis in a single numerical value that measures the closeness in the l2 - norm of the test sample to the mean prototype of each class. A negative DI indicates normobiosis, whereas a positive DI indicates dysbiosis. For a threshold of 0, the DI based on the combined dataset achieved 74% sensitivity and 95% specificity to separate healthy and CE dogs. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Effects of professional oral health care on elderly: randomized trial.

    Science.gov (United States)

    Morino, T; Ookawa, K; Haruta, N; Hagiwara, Y; Seki, M

    2014-11-01

    To better understand the role of the professional oral health care for elderly in improving geriatric oral health, the effects of short-term professional oral health care (once per week for 1 month) on oral microbiological parameters were assessed. Parallel, open-labelled, randomize-controlled trial was undertaken in a nursing home for elderly in Shizuoka, Japan. Thirty-four dentate elderly over 74 years were randomly assigned from ID number to the intervention (17/34) and control (17/34) groups. The outcomes were changes in oral microbiological parameters (number of bacteria in unstimulated saliva; whole bacteria, Streptococcus, Fusobacterium and Prevotella: opportunistic pathogens detection: and index of oral hygiene evaluation [Dental Plaque Index, DPI]) within the intervention period. Each parameter was evaluated at before and after intervention period. Four elderly were lost from mortality (1), bone fracture (1), refused to participate (1) and multi-antibiotics usage (1). Finally, 30 elderly were analysed (14/intervention and 16/control). At baseline, no difference was found between the control and intervention groups. After the intervention period, the percentage of Streptococcus species increased significantly in the intervention group (Intervention, 86% [12/14]; Control, 50% [8/16]: Fisher's, right-tailed, P oral health care can improve oral conditions in the elderly. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  13. Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

    Energy Technology Data Exchange (ETDEWEB)

    Gangi Setty, Thanuja [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India); Cho, Christine [Carver College of Medicine, University of Iowa, Iowa City, IA 52242-1109 (United States); Govindappa, Sowmya [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India); Apicella, Michael A. [Carver College of Medicine, University of Iowa, Iowa City, IA 52242-1109 (United States); Ramaswamy, S., E-mail: ramas@instem.res.in [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India)

    2014-07-01

    Structure–function studies of sialic acid-binding proteins from F. nucleatum, P. multocida, V. cholerae and H. influenzae reveal a conserved network of hydrogen bonds involved in conformational change on ligand binding. Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states.

  14. Removal and prevention of dental plaque with d-tagatose.

    Science.gov (United States)

    Lu, Y; Levin, G V

    2002-08-01

    Dental plaque develops when early bacterial colonizers adhere to the acquired pellicle (saliva-derived proteinous coating on the tooth surface) followed by adhesion of late interspecies colonizers to form this type of biofilm (coaggregation). In developing a d-tagatose-based toothpaste, we examined 15 oral isolates, including both early colonizers (Streptococcus and Actinomyces) and late colonizers (Fusobacterium, Porphyromonas, Prevotella, Veillonella, Capnocytophaga, and Actinobacillus), and tested them for their ability to coaggregate with each other. We then tested the ability of d-tagatose to reverse any such coaggregations. Coaggregation was examined visually and scored by using a system ranging from 0, for no visible coaggregation to 4, for maximum coaggregation. d-Tagatose, at a concentration of less than 750 mm, completely reversed the coaggregation of 17 (60%) of 28 strongly coaggregating pairs (coaggregation score = 2 or higher) tested. In contrast, d-sorbitol had little reversal effect. d-Tagatose-sensitive coaggregations were d-galactose-reversible as well. d-Tagatose acted on both early and late colonizers; both groups, especially the late colonizers, were frequently involved in periodontal diseases. Thus, d-tagatose has the potential for preventing and removing plaque development and for altering the subgingival microbiota. These effective qualities offer conservative control of gingival and periodontal disease.

  15. Lemierre's syndrome

    DEFF Research Database (Denmark)

    Johannesen, Katrine M; Bodtger, Uffe

    2016-01-01

    This is a systematic review of cases with Lemierre's syndrome (LS) in the past 5 years. LS is characterized by sepsis often evolving after a sore throat or tonsillitis and then complicated by various septic emboli and thrombosis of the internal jugular vein. Symptoms include sepsis, pain, and/or ...... LS in this day and age appears to be low, however the syndrome is difficult to recognize, and still requires the full attention of the clinician.......This is a systematic review of cases with Lemierre's syndrome (LS) in the past 5 years. LS is characterized by sepsis often evolving after a sore throat or tonsillitis and then complicated by various septic emboli and thrombosis of the internal jugular vein. Symptoms include sepsis, pain, and....../or swelling in the throat or neck, as well as respiratory symptoms. Laboratory findings show elevated infectious parameters and radiological findings show thrombosis of the internal jugular vein and emboli in the lungs or other organs. The syndrome is often associated with an infection with Fusobacterium...

  16. Microbiological, lipid and immunological profiles in children with gingivitis and type 1 diabetes mellitus

    Science.gov (United States)

    DUQUE, Cristiane; JOÃO, Mariana Ferreira Dib; CAMARGO, Gabriela Alessandra da Cruz Galhardo; TEIXEIRA, Gláucia Schuindt; MACHADO, Thamiris Santana; AZEVEDO, Rebeca de Souza; MARIANO, Flávia Sammartino; COLOMBO, Natália Helena; VIZOTO, Natália Leal; MATTOS-GRANER, Renata de Oliveira

    2017-01-01

    Abstract Objective The aim of this study was to compare the prevalence of periodontal pathogens, systemic inflammatory mediators and lipid profiles in type 1 diabetes children (DM) with those observed in children without diabetes (NDM), both with gingivitis. Material and methods Twenty-four DM children and twenty-seven NDM controls were evaluated. The periodontal status, glycemic and lipid profiles were determined for both groups. Subgingival samples of periodontal sites were collected to determine the prevalence of periodontal microorganisms by PCR. Blood samples were collected for IL-1-β, TNF-α and IL-6 analysis using ELISA kits. Results Periodontal conditions of DM and NDM patients were similar, without statistical differences in periodontal indices. When considering patients with gingivitis, all lipid parameters evaluated were highest in the DM group; Capnocytophaga sputigena and Capnocytophaga ochracea were more prevalent in the periodontal sites of DM children. “Red complex” bacteria were detected in few sites of DM and NDM groups. Fusobacterium nucleatum and Campylobacter rectus were frequently found in both groups. Similar levels of IL-1-β, TNF-α and IL-6 were detected in DM and NDM children. Conclusion Clinical and immunological profiles are similar between DM and NDM children. The presence of Capnocytophaga sputigena and Capnocytophaga ochracea were associated with gingivitis in DM children. PMID:28403363

  17. Microbiomes associated with bovine periodontitis and oral health.

    Science.gov (United States)

    Borsanelli, Ana C; Lappin, David F; Viora, Lorenzo; Bennett, David; Dutra, Iveraldo S; Brandt, Bernd W; Riggio, Marcello P

    2018-05-01

    Periodontitis is an infectious polymicrobial, immuno-inflammatory disease of multifactorial aetiology that has an impact on the health, production and welfare of ruminants. The objective of the present study was to determine the microbial profiles present in the gingival sulcus of cattle considered periodontally healthy and in the periodontal pocket of animals with periodontitis lesions using high-throughput bacterial 16S rRNA gene sequencing. Subgingival biofilm samples were collected from 40 cattle with periodontitis and 38 periodontally healthy animals. In total, 1923 OTUs were identified and classified into 395 genera or higher taxa. Microbial profiles in health differed significantly from periodontitis in their composition (p PERMANOVA) but no statistically significant differences were observed in the diversity of healthy and periodontitis microbiomes. The most prevalent taxa in health were Pseudomonas, Burkholderia and Actinobacteria, whereas in disease these were Prevotella, Fusobacterium and Porphyromonas. The most discriminative taxa in health were Gastranaerophilales, Planifilum and Burkholderia, and in disease these were Elusimicrobia, Synergistes and Propionivibrio. In conclusion, statistically significant difference exists between the microbiome in bovine oral health and periodontitis, with populations showing 72.6% dissimilarity. The diversity of the bacteria found in health and periodontitis were similar and bacteria recognised as periodontal pathogens showed increased abundance in disease. In this context, the main components of bacterial homeostasis in the biofilm of healthy sites and of dysbiosis in periodontal lesions provide unprecedented indicators for the evolution of knowledge about bovine periodontitis. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Dysbiosis of the microbiome in gastric carcinogenesis.

    Science.gov (United States)

    Castaño-Rodríguez, Natalia; Goh, Khean-Lee; Fock, Kwong Ming; Mitchell, Hazel M; Kaakoush, Nadeem O

    2017-11-21

    The gastric microbiome has been proposed as an etiological factor in gastric carcinogenesis. We compared the gastric microbiota in subjects presenting with gastric cancer (GC, n = 12) and controls (functional dyspepsia (FD), n = 20) from a high GC risk population in Singapore and Malaysia. cDNA from 16S rRNA transcripts were amplified (515F-806R) and sequenced using Illumina MiSeq 2 × 250 bp chemistry. Increased richness and phylogenetic diversity but not Shannon's diversity was found in GC as compared to controls. nMDS clustered GC and FD subjects separately, with PERMANOVA confirming a significant difference between the groups. H. pylori serological status had a significant impact on gastric microbiome α-diversity and composition. Several bacterial taxa were enriched in GC, including Lactococcus, Veilonella, and Fusobacteriaceae (Fusobacterium and Leptotrichia). Prediction of bacterial metabolic contribution indicated that serological status had a significant impact on metabolic function, while carbohydrate digestion and pathways were enriched in GC. Our findings highlight three mechanisms of interest in GC, including enrichment of pro-inflammatory oral bacterial species, increased abundance of lactic acid producing bacteria, and enrichment of short chain fatty acid production pathways.

  19. Enteral tube feeding alters the oral indigenous microbiota in elderly adults.

    Science.gov (United States)

    Takeshita, Toru; Yasui, Masaki; Tomioka, Mikiko; Nakano, Yoshio; Shimazaki, Yoshihiro; Yamashita, Yoshihisa

    2011-10-01

    Enteral tube feeding is widely used to maintain nutrition for elderly adults with eating difficulties, but its long-term use alters the environment of the oral ecosystem. This study characterized the tongue microbiota of tube-fed elderly adults by analyzing the 16S rRNA gene. The terminal restriction fragment length polymorphism (T-RFLP) profiles of 44 tube-fed subjects were compared with those of 54 subjects fed orally (average age, 86.4 ± 6.9 years). Bar-coded pyrosequencing data were also obtained for a subset of the subjects from each group (15 tube-fed subjects and 16 subjects fed orally). The T-RFLP profiles demonstrated that the microbiota of the tube-fed subjects was distinct from that of the subjects fed orally (permutational multivariate analysis of variance [perMANOVA], P < 0.001). The pyrosequencing data revealed that 22 bacterial genera, including Corynebacterium, Peptostreptococcus, and Fusobacterium, were significantly more predominant in tube-fed subjects, whereas the dominant genera in the subjects fed orally, such as Streptococcus and Veillonella, were present in much lower proportions. Opportunistic pathogens rarely detected in the normal oral microbiota, such as Corynebacterium striatum and Streptococcus agalactiae, were often found in high proportions in tube-fed subjects. The oral indigenous microbiota is disrupted by the use of enteral feeding, allowing health-threatening bacteria to thrive.

  20. Periodontal microbioma and rheumatoid arthritis: The role of Porhyromonas gingivalis.

    Science.gov (United States)

    Azzi, L; Rania, S; Vinci, R; Spadari, F; Croveri, F; Scognamiglio, C; Farronato, D; Tettamanti, L; Tagliabue, A; Silvestre-Rangil, J; Bellintani, C

    2017-01-01

    Rheumatoid Arthritis is a disease, which can be described as an autoimmune response after molecular mimicry caused by infective agents. The current study aims at evaluating the correlation between Rhematoid Arthritis (RA) and Periodontal Disease (PD), with special attention to the microbioma detected in the gums. Thirty-four patients with RD were recruited into the current study. Among rheumatic parameters, Rheumatoid Factor (RF), anti-citrullinated protein antibody (CCP), HLA-BDR1 and DAS28 were collected. A dental clinician evaluated the periodontal screening record (PSR). Afterwards, 1 paper cone was inserted for 30 seconds into the gingival sulcus then sent to the laboratory for evaluation. Quantitative PCR of 16S rRNA genes was performed with the hydrolysis probes method to identify and evaluate the amount Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, Fusobacterium nucleatum and Campylobacter rectus. There were no statistical differences in the composition of oral microbioma between PSR groups. There were no statistical significant differences between bacterial loads and serum values. On the contrary, a positive correlation was found between the presence of Porphyromonas gingivalis in periodontal pockets on one side and RF and CCP on the other. Therefore, the presence of Porhyromonas gingivalis in periodontal pockets is associated to RA inflammatory indices.

  1. Mining the Proteome of subsp. ATCC 25586 for Potential Therapeutics Discovery: An Approach

    Directory of Open Access Journals (Sweden)

    Abdul Musaweer Habib

    2016-12-01

    Full Text Available The plethora of genome sequence information of bacteria in recent times has ushered in many novel strategies for antibacterial drug discovery and facilitated medical science to take up the challenge of the increasing resistance of pathogenic bacteria to current antibiotics. In this study, we adopted subtractive genomics approach to analyze the whole genome sequence of the Fusobacterium nucleatum, a human oral pathogen having association with colorectal cancer. Our study divulged 1,499 proteins of F. nucleatum, which have no homolog's in human genome. These proteins were subjected to screening further by using the Database of Essential Genes (DEG that resulted in the identification of 32 vitally important proteins for the bacterium. Subsequent analysis of the identified pivotal proteins, using the Kyoto Encyclopedia of Genes and Genomes (KEGG Automated Annotation Server (KAAS resulted in sorting 3 key enzymes of F. nucleatum that may be good candidates as potential drug targets, since they are unique for the bacterium and absent in humans. In addition, we have demonstrated the three dimensional structure of these three proteins. Finally, determination of ligand binding sites of the 2 key proteins as well as screening for functional inhibitors that best fitted with the ligands sites were conducted to discover effective novel therapeutic compounds against F. nucleatum.

  2. Periodontopathic microorganisms in peripheric blood after scaling and root planing.

    Science.gov (United States)

    Lafaurie, Gloria Inés; Mayorga-Fayad, Isabel; Torres, María Fernanda; Castillo, Diana Marcela; Aya, Maria Rosario; Barón, Alexandra; Hurtado, Paola Andrea

    2007-10-01

    The objective of this study was to evaluate the frequency of periodontopathic and other subgingival anaerobic and facultative bacteria in the bloodstream following scaling and root planing (SRP). Forty-two patients with severe generalized chronic periodontitis (GChP) and generalized aggressive periodontitis (GAgP) were included in the study. Four samples of peripheric blood were drawn from the cubital vein at different times: Pre-treatment: immediately before the SRP procedure (T1), immediately after treatment (T2), 15 min. post-treatment (T3) and 30 min. post-treatment (T4). In order to identify the presence of microorganisms in blood, subcultures were conducted under anaerobic conditions. 80.9% of the patients presented positive cultures after SRP and it occurred more frequently immediately after treatment; however, 19% of the patients still had microorganisms in the bloodstream 30 min. after the procedure. The periodontopathic microorganisms more frequently identified were Porphyromonas gingivalis and Micromonas micros. Campylobacter spp., Eikenella corrodens, Tannerella forsythensis, Fusobacterium spp. and Prevotella intermedia were isolated less often. Actinomyces spp. were also found frequently during bacteraemia after SRP. SRP induced bacteraemia associated with anaerobic bacteria, especially in patients with periodontal disease.

  3. [Relationships between the enrichment of ETBF, Fn, Hp in intestinal and colorectal cancer].

    Science.gov (United States)

    Zhang, J; Lu, X L; Zhao, G; Shi, H T; Geng, Y; Zhong, W T; Dong, L

    2018-02-23

    Objective: To explore relationships between the enrichment of ETBF, Fn, Hp in feces, tissues and colorectal cancer. Methods: Feces, lesion tissue and adjacent tissue from 24 patients with colorectal cancer and 31 patients with adenomas were collected, and we collected Feces and tissue of 20 healthy control persons. Then the copy numbers of enterotoxigenic B. fragilis (ETBF), Fusobacterium nucleatum (Fn) and Helicobacter pylori (Hp) were determined by quantitative real-time PCR. Immunohistochemical method was used to examine the expression intensity of EGFR and p53, and the relationships between different expression intensity of EGFR, p53 and the numbers of three bacterias. Results: In the feces, copy numbers of ETBF and Fn were as follous: colorectal cancer group>adenomas group>healthy control group ( P healthy control group ( P healthy control group ( P adenomas group>healthy control group ( P healthy control group ( P healthy control group ( P bacteria in the lesion tissue and the adjacent tissue had no significant difference. This happened both in colorectal cancer group and adenomas group. The different expression intensity of EGFR, p53 and the number of three bacteria showed no obviously statistical correlation( P >0.05). Conclusion: Adenomatous polyp and colorectal cancer patients show high enrichment of ETBF, Fn and Hp in both feces and tissues. ETBF, Fn and Hp probably contribute to the development of adenomatous polyp and colorectal cancer. Trial registration Chinese Clinical Trial Registry, ChiCTR-BOC-17012509.

  4. Periodontal pathogen levels in adolescents before, during, and after fixed orthodontic appliance therapy.

    Science.gov (United States)

    Thornberg, Michelle J; Riolo, Christopher S; Bayirli, Burcu; Riolo, Michael L; Van Tubergen, Elizabeth A; Kulbersh, Richard

    2009-01-01

    This purpose of this study was to document and investigate changes in periodontal pathogen levels before, during, and after orthodontic treatment in adolescents. DNA gene probe analysis was used to quantify the levels of 8 periodontal pathogens before, during, and after treatment with fixed orthodontic appliances in 190 concurrently treated adolescent orthodontic patients. The 8 pathogens examined were Actinobacillus actinomycetemcomitans (AA), Porphyromonas gingivalis (PG), Prevotella intermedia (PI), Tannerella forsythia (TF), Eikenella corrodens (EC), Fusobacterium nucleatum (FN), Treponema denticola (TD), and Campylobacter rectus (CR). Chi-square tests were used to determine whether the percentages of subjects with high counts significantly changed over time. Logistic regression analyses were also performed to derive the relative risk of higher counts of pathogenic bacteria with fixed appliances at the various time intervals studied. For 6 (PI, TF, EC, FN, TD, CR) of the 8 pathogens, the percentages of subjects with high pathogen counts increased significantly after 6 months of fixed appliance treatment, but these returned to pretreatment levels by 12 months of orthodontic treatment. No pathogen level was significantly higher after 12 months of orthodontic treatment, and orthodontic treatment was found to be significantly protective for half of the pathogens (EC, FN, TD, CR) posttreatment. Orthodontic treatment with fixed appliances does not increase the risk of high levels of these periodontal pathogens.

  5. Focus on periodontal disease and colorectal carcinoma.

    Science.gov (United States)

    Lauritano, D; Sbordone, L; Nardone, M; Iapichino, A; Scapoli, L; Carinci, F

    2017-01-01

    Diagnosis of focal disease, the theory that the human oral microbial (HOM) could affect the onset and development of systemic diseases, was very popular in the past, but the lack of scientific evidence has led to the abandonment of this idea. Interestingly, increasing evidence over the past 3 or so decades suggests that HOM can indeed serve as a reservoir for systemic dissemination of pathogenic bacteria and their toxins in distant body sites, favouring the developments of malignant tumours. Malignant tumours are complex communities of oncogenically transformed cells with aberrant genomes, associated non-neoplastic cells including immune and stromal cells, and sometimes HOM, including bacteria and viruses. Recent data suggest that HOM and periodontal disease play an active role in the pathogenesis of colorectal cancer, in fact HOM has been found within the colorectal cancer microenvironment, and the composition of the HOM was different from that of adjacent non-neoplastic tissue. An association of fusobacterium nucleatum with the colonic mucosa of colorectal cancer has been proven. Several questions thus arise. Is periodontal disease a risk factor for colorectal carcinoma? Given the connectivity of the digestive tract, could fusubacterium nucleatum or other HOM be involved in additional gastrointestinal disorders? Furthermore, based on the "mobility" of Fusubacterium nucleatum and the omnipresence of cadherins, could this organism be involved in cancers beyond the gastrointestinal tract? Answers to these questions will shed new lights on the role of the HOM in onset of diseases.

  6. [Microbiota of urine and vagina of healthy postmenopausal women (a pilot study)].

    Science.gov (United States)

    Naboka, Yu L; Rymashevsky, A N; Kogan, M I; Gudima, I A; Borovleva, O A; Jalagonia, K T; Zarutskiy, S A

    2016-02-01

    Studying microbiota of different urogenital tract habitats in healthy postmenopausal women is of practical importance in deciding on the appropriateness of correction of dysbiotic disorders. The aim of this study was to examine the vaginal and urine microbiota of healthy postmenopausal women. The study included 20 healthy postmenopausal women (mean age 59,0+/-2,1 years). Duration of menopause in all subjects was more than 8 years. Bacteriological testing of urine and vaginal specimen was carried out on the extended media (15) for cultivating facultative anaerobic bacteria (FAB) and nonclostridial anaerobic bacteria (NAB) and included PCR of midstream morning urine. Among FAB in the urine and vagina dominated coagulase-negative staphylococci and NAB. Bacterial patterns of studied habitats turned out to be similar in many respects. In the urine Megasphaera spp., Veillonella spp., Prevotella spp., Mobiluncus spp., Fusobacterium spp. were found, whereas in the vagina these microorganisms were not present. Cluster analysis revealed no significant differences in the concentration of the same microorganisms isolated from the urine and vagina. When comparing the frequency of microorganism detection in urine by bacteriological method and by PCR, bacterial patterns were identical in 56% of cases.

  7. Modulation of microbial consortia enriched from different polluted environments during petroleum biodegradation.

    Science.gov (United States)

    Omrani, Rahma; Spini, Giulia; Puglisi, Edoardo; Saidane, Dalila

    2018-04-01

    Environmental microbial communities are key players in the bioremediation of hydrocarbon pollutants. Here we assessed changes in bacterial abundance and diversity during the degradation of Tunisian Zarzatine oil by four indigenous bacterial consortia enriched from a petroleum station soil, a refinery reservoir soil, a harbor sediment and seawater. The four consortia were found to efficiently degrade up to 92.0% of total petroleum hydrocarbons after 2 months of incubation. Illumina 16S rRNA gene sequencing revealed that the consortia enriched from soil and sediments were dominated by species belonging to Pseudomonas and Acinetobacter genera, while in the seawater-derived consortia Dietzia, Fusobacterium and Mycoplana emerged as dominant genera. We identified a number of species whose relative abundances bloomed from small to high percentages: Dietzia daqingensis in the seawater microcosms, and three OTUs classified as Acinetobacter venetianus in all two soils and sediment derived microcosms. Functional analyses on degrading genes were conducted by comparing PCR results of the degrading genes alkB, ndoB, cat23, xylA and nidA1 with inferences obtained by PICRUSt analysis of 16S amplicon data: the two data sets were partly in agreement and suggest a relationship between the catabolic genes detected and the rate of biodegradation obtained. The work provides detailed insights about the modulation of bacterial communities involved in petroleum biodegradation and can provide useful information for in situ bioremediation of oil-related pollution.

  8. Antimicrobial activity of isothiocyanates (ITCs) extracted from horseradish (Armoracia rusticana) root against oral microorganisms.

    Science.gov (United States)

    Park, Ho-Won; Choi, Kyu-Duck; Shin, Il-Shik

    2013-01-01

    The antimicrobial activity of isothiocyanates (ITCs) extracted from horseradish root was investigated against oral microorganisms: 6 strains of facultative anaerobic bacteria, Streptococcus mutans, Streptococcus sobrinus, Lactobacillus casei, Staphylococcus aureus, Enterococcus faecalis and Aggregatibacter actinomycetemcomitans; one strain of yeast, Candida albicans, and 3 strains of anaerobic bacteria, Fusobacterium nucleatum, Prevotella nigrescens, and Clostridium perfringens. The ITCs extracted from horseradish root showed antimicrobial activity against all oral microorganisms by the paper disk method. The minimum bactericidal concentration (MBC) of the ITCs extracted from horseradish root ranged from 1.25 to 5.00 mg/ml against 6 strains of facultative anaerobic bacteria and one strain of yeast, and 4.17 to 16.67 mg/ml against 3 strains of anaerobic bacteria. The ITCs extracted from horseradish root showed the strongest antimicrobial activity, with a MBC of 1.25 mg/ml, against C. albicans among facultative microorganisms, and 4.17 mg/ml against F. nucleatum among anaerobic bacteria. These results suggest that the ITCs extracted from horseradish root may be a candidate for use as an antimicrobial agent against oral microorganisms.

  9. Dog and human inflammatory bowel disease rely on overlapping yet distinct dysbiosis networks.

    Science.gov (United States)

    Vázquez-Baeza, Yoshiki; Hyde, Embriette R; Suchodolski, Jan S; Knight, Rob

    2016-10-03

    Inflammatory bowel disease (IBD) is an autoimmune condition that is difficult to diagnose, and animal models of this disease have questionable human relevance 1 . Here, we show that the dysbiosis network underlying IBD in dogs differs from that in humans, with some bacteria such as Fusobacterium switching roles between the two species (as Bacteroides fragilis switches roles between humans and mice) 2 . For example, a dysbiosis index trained on humans fails when applied to dogs, but a dog-specific dysbiosis index achieves high correlations with the overall dog microbial community diversity patterns. In addition, a random forest classifier trained on dog-specific samples achieves high discriminatory power, even when using stool samples rather than the mucosal biopsies required for high discriminatory power in humans 2 . These relationships were not detected in previously published dog IBD data sets due to their limited sample size and statistical power 3 . Taken together, these results reveal the need to train host-specific dysbiosis networks and point the way towards a generalized understanding of IBD across different mammalian models.

  10. In Vitro Evaluation of Planktonic Growth on Experimental Cement-Retained Titanium Surfaces

    Science.gov (United States)

    Balci, Nur; Cakan, Umut; Aksu, Burak; Akgul, Oncu; Ulger, Nurver

    2016-01-01

    Background The purpose of this study was to compare the effects of selected cements, or their combination with titanium, on the growth of two periodontopathic bacteria: Prevotella intermedia (Pi) and Fusobacterium nucleatum (Fn). Material/Methods This study was comprised of several experimental groups: 1) Dental luting cements (glass ionomer cement, methacrylate-based resin cement, zinc-oxide eugenol cement, eugenol-free zinc oxide cement; 2) titanium discs; and 3) titanium combination cement discs. The disks were submerged in bacterial suspensions of either Fn or Pi. Planktonic bacterial growth within the test media was measured by determining the optical density of the cultures (OD600). Mean and standard deviations were calculated for planktonic growth from three separate experiments. Results Intergroup comparison of all experimental groups revealed increased growth of Pi associated with cement-titanium specimens in comparison with cement specimens. Regarding the comparison of all groups for Fn, there was an increased amount of bacterial growth in cement-titanium specimens although the increase was not statistically significant. Conclusions The combination of cement with titanium may exacerbate the bacterial growth capacity of Pi and Fn in contrast to their sole effect. PMID:27058704

  11. Preliminary Comparison of Oral and Intestinal Human Microbiota in Patients with Colorectal Cancer: A Pilot Study

    Directory of Open Access Journals (Sweden)

    Edda Russo

    2018-01-01

    Full Text Available In this study Next-Generation Sequencing (NGS was used to analyze and compare human microbiota from three different compartments, i.e., saliva, feces, and cancer tissue (CT, of a selected cohort of 10 Italian patients with colorectal cancer (CRC vs. 10 healthy controls (saliva and feces. Furthermore, the Fusobacterium nucleatum abundance in the same body site was investigated through real-time quantitative polymerase chain reaction (qPCR to assess the association with CRC. Differences in bacterial composition, F. nucleatum abundance in healthy controls vs. CRC patients, and the association of F. nucleatum with clinical parameters were observed. Taxonomic analysis based on 16S rRNA gene, revealed the presence of three main bacterial phyla, which includes about 80% of reads: Firmicutes (39.18%, Bacteroidetes (30.36%, and Proteobacteria (10.65%. The results highlighted the presence of different bacterial compositions; in particular, the fecal samples of CRC patients seemed to be enriched with Bacteroidetes, whereas in the fecal samples of healthy controls Firmicutes were one of the major phyla detected though these differences were not statistically significant. The CT samples showed the highest alpha diversity values. These results emphasize a different taxonomic composition of feces from CRC compared to healthy controls. Despite the low number of samples included in the study, these results suggest the importance of microbiota in the CRC progression and could pave the way to the development of therapeutic interventions and novel microbial-related diagnostic tools in CRC patients.

  12. Indigenous bacteria and bacterial metabolic products in the gastrointestinal tract of broiler chickens.

    Science.gov (United States)

    Rehman, Habib Ur; Vahjen, Wilfried; Awad, Wageha A; Zentek, Jürgen

    2007-10-01

    The gastrointestinal tract is a dynamic ecosystem containing a complex microbial community. In this paper, the indigenous intestinal bacteria and the microbial fermentation profile particularly short chain fatty acids (SCFA), lactate, and ammonia concentrations are reviewed. The intestinal bacterial composition changes with age. The bacterial density of the small intestine increases with age and comprises of lactobacilli, streptococci, enterobacteria, fusobacteria and eubacteria. Strict anaerobes (anaerobic gram-positive cocci, Eubacterium spp., Clostridium spp., Lactobacillus spp., Fusobacterium spp. and Bacteroides) are predominating caecal bacteria in young broilers. Data from culture-based studies showed that bifidobacteria could not be isolated from young birds, but were recovered from four-week-old broilers. Caecal lactobacilli accounted for 1.5-24% of the caecal bacteria. Gene sequencing of caecal DNA extracts showed that the majority of bacteria belonged to Clostridiaceae. Intestinal bacterial community is influenced by the dietary ingredients, nutrient levels and physical structure of feed. SCFA and other metabolic products are affected by diet formulation and age. Additional studies are required to know the bacterial metabolic activities together with the community analysis of the intestinal bacteria. Feed composition and processing have great potential to influence the activities of intestinal bacteria towards a desired direction in order to support animal health, well-being and microbial safety of broiler meat.

  13. Microbiota associated with chronic osteomyelitis of the jaws

    Directory of Open Access Journals (Sweden)

    Elerson Gaetti-Jardim Júnior

    2010-12-01

    Full Text Available Chronic osteomyelitis of maxilla and mandible is rare in industrialized countries and its occurrence in developing countries is associated with trauma and surgery, and its microbial etiology has not been studied thoroughly. The aim of this investigation was to evaluate the microbiota associated with osteomyelitis of mandible or maxilla from some Brazilian patients. After clinical and radiographic evaluation, samples of bone sequestra, purulent secretion, and biopsies of granulomatous tissues from twenty-two patients with chronic osteomyelitis of mandible and maxilla were cultivated and submitted for pathogen detection by using a PCR method. Each patient harbored a single lesion. Bacterial isolation was performed on fastidious anaerobe agar supplemented with hemin, menadione and horse blood for anaerobes; and on tryptic soy agar supplemented with yeast extract and horse blood for facultative bacteria and aerobes. Plates were incubated in anaerobiosis and aerobiosis, at 37ºC for 14 and 3 days, respectively. Bacteria were cultivated from twelve patient samples; and genera Actinomyces, Fusobacterium, Parvimonas, and Staphylococcus were the most frequent. By PCR, bacterial DNA was detected from sixteen patient samples. The results suggest that cases of chronic osteomyelitis of the jaws are usually mixed anaerobic infections, reinforcing the concept that osteomyelitis of the jaws are mainly related to microorganisms from the oral environment, and periapical and periodontal infections may act as predisposing factors.

  14. The Effects of Antimicrobial Peptide Nal-P-113 on Inhibiting Periodontal Pathogens and Improving Periodontal Status

    Directory of Open Access Journals (Sweden)

    Hongyan Wang

    2018-01-01

    Full Text Available Periodontal disease consists of chronic gingival inflammation characterized by both degradation of the periodontal connective tissue and alveolar bone loss. Drug therapy is used as an auxiliary treatment method in severe chronic periodontitis, aggressive periodontitis, and periodontitis-associated systemic disease. Nal-P-113, a modified antimicrobial peptide, specifically replaces the histidine residues of P-113 with the bulky amino acid β-naphthylalanine, and our previous studies have verified that this novel peptide is not toxic to the human body within a certain concentration range. The objective of the present study was to evaluate the effect of Nal-P-113 on periodontal pathogens and periodontal status in clinical studies. In a split-mouth clinical trial, the pocket depth and bleeding index values tended to decrease in the experimental group compared with those in the control group. SEM results verified that Nal-P-113 restrained the maturation of plaque. Based on real-time polymerase chain reaction, the levels of Fusobacterium nucleatum, Streptococcus gordonii, Treponema denticola, and Porphyromonas gingivalis in subgingival plaque were decreased when the subjects were given Nal-P-113. Bacterial growth curve analysis and a biofilm susceptibility assay verified that Nal-P-113 at a concentration of 20 μg/mL restrained the growth of S. gordonii, F. nucleatum, and P. gingivalis and biofilm formation. Therefore, Nal-P-113 effectively reduces periodontal pathogens and ameliorates periodontal status.

  15. Effect of green tea catechin, a local drug delivery system as an adjunct to scaling and root planing in chronic periodontitis patients: A clinicomicrobiological study

    Science.gov (United States)

    Kudva, Praveen; Tabasum, Syeda Tawkhira; Shekhawat, Nirmal Kanwar

    2011-01-01

    Background: Evaluate the adjunctive use of locally delivered green tea catechin with scaling and root planing, as compared to scaling and root planing alone in the management of chronic periodontitis. Materials and Methods: Fourteen patients with two sites in the contralateral quadrants with probing pocket depth of 5–8mm were selected. Each of the sites was assessed for the plaque index, gingival index, and probing pocket depth at baseline and 21 days and for microbiological analysis at baseline, 1 week and 21 days. Test sites received scaling and root planing along with green tea catechin strips and control sites received scaling and root planning alone. Results: The result showed intercomparison of the plaque index and gingival index for test and control groups at 21 days was not significant with P>0.05, whereas the probing depth at 21 days was significant with P<0.001. Intercomparison between microbial results demonstrated a considerable reduction of occurrence of Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Fusobacterium species and Capnocytophaga in test. Conclusion: Green tea catechin local delivery along with scaling and root planing is more effective than scaling and root planing alone. PMID:21772720

  16. Evaluation of the Vitek 2 ANC card for identification of clinical isolates of anaerobic bacteria.

    Science.gov (United States)

    Lee, E H L; Degener, J E; Welling, G W; Veloo, A C M

    2011-05-01

    An evaluation of the Vitek 2 ANC card (bioMérieux, Marcy l'Etoile, France) was performed with 301 anaerobic isolates. Each strain was identified by 16S rRNA gene sequencing, which is considered to be the reference method. The Vitek 2 ANC card correctly identified 239 (79.4%) of the 301 clinical isolates to the genus level, including 100 species that were not represented in the database. Correct species identification was obtained for 60.1% (181/301) of the clinical isolates. For the isolates not identified to the species level, a correct genus identification was obtained for 47.0% of them (47/100), and 16 were accurately designated not identified. Although the Vitek 2 ANC card allows the rapid and acceptable identification of the most common clinically important anaerobic bacteria within 6 h, improvement is required for the identification of members of the genera Fusobacterium, Prevotella, and Actinomyces and certain Gram-positive anaerobic cocci (GPAC).

  17. The influence of orthodontic fixed appliances on the oral microbiota: A systematic review

    Directory of Open Access Journals (Sweden)

    Amanda Osório Ayres de Freitas

    2014-04-01

    Full Text Available Objective: To investigate whether there is scientific evidence to support the hypothesis that the presence of orthodontic fixed appliances influences the oral microbiota. Methods: The search for articles was conducted in PubMed; ISI Web of Knowledge and Ovid databases, including articles published in English until May 17th, 2012. They should report human observational studies presenting the following keywords: "fixed orthodontic appliance" AND "microbiological colonization"; OR "periodontal pathogens"; OR "Streptococcus"; OR "Lactobacillus"; OR "Candida"; OR "Tannerella forsythia"; OR "Treponema denticola"; OR "Fusobacterium nucleatum"; OR "Actimomyces actinomycetemcomitans"; OR "Prevotella intermedia", OR "Prevotella nigrescens"; OR "Porphyromonas gingivalis". Articles were previously selected by title and abstract. Articles that met the inclusion criteria were analyzed and classified as having low, moderate or high methodology quality. A new detailed checklist for quality assessment was developed based on the information required for applicable data extraction for reviews. The study design, sample, follow-up period, collection and microbial analysis methods, statistical treatment, results and discussion were assessed. Results: The initial search retrieved 305 articles of which 33 articles were selected by title and abstract. After full-text reading, 8 articles met the inclusion criteria, out of which 4 articles were classified as having low and 4 as moderate methodological quality. The moderate methodological quality studies were included in the systematic review. Conclusions: The literature revealed moderate evidence that the presence of fixed appliances influences the quantity and quality of oral microbiota.

  18. Microcalorimetric determination of the effects of amoxicillin, metronidazole, and their combination on in vitro biofilm.

    Science.gov (United States)

    Astasov-Frauenhoffer, Monika; Braissant, Olivier; Hauser-Gerspach, Irmgard; Weiger, Roland; Walter, Clemens; Zitzmann, Nicola U; Waltimo, Tuomas

    2014-02-01

    The mechanism of action of adjuvant antibiotic therapy in the treatment of peri-implantitis is not well understood. The aim of this study is to investigate antibiotic susceptibility of an in vitro biofilm by isothermal microcalorimetry (IMC). Titanium disks containing a 72-hour three-species biofilm (Streptococcus sanguinis DSM20068, Fusobacterium nucleatum ATCC10953, and Porphyromonas gingivalis DSM20709) were placed in a series of IMC ampoules with nutrient agar supplemented with increasing concentrations of amoxicillin, metronidazole, or their combination and incubated anaerobically for 10 days. Lag time and maximum growth rate were determined from continuous heat-flow recordings of metabolic activity. To validate the IMC biofilm results, adherent S. sanguinis and P. gingivalis were incubated anaerobically in medium supplemented with antibiotics at 37°C for 24 hours, and their vitality was determined by live/dead staining, conventional culturing, and IMC. In all biofilm samples incubated with antibiotics, a prolonged lag phase was observed compared with controls (P live/dead staining and conventional culturing. IMC gives new evidence about antibiotic effects on oral biofilms and is more informative than conventional culture and live/dead assays. The combination of antibiotics was found to be more efficient than metronidazole alone; however, only minor differences in growth inhibition were detected compared with amoxicillin alone.

  19. Comparative cytotoxicity of periodontal bacteria

    International Nuclear Information System (INIS)

    Stevens, R.H.; Hammond, B.F.

    1988-01-01

    The direct cytotoxicity of sonic extracts (SE) from nine periodontal bacteria for human gingival fibroblasts (HGF) was compared. Equivalent dosages (in terms of protein concentration) of SE were used to challenge HGF cultures. The cytotoxic potential of each SE was assessed by its ability to (1) inhibit HGF proliferation, as measured by direct cell counts; (2) inhibit 3H-thymidine incorporation in HGF cultures; or (3) cause morphological alterations of the cells in challenged cultures. The highest concentration (500 micrograms SE protein/ml) of any of the SEs used to challenge the cells was found to be markedly inhibitory to the HGFs by all three of the criteria of cytotoxicity. At the lowest dosage tested (50 micrograms SE protein/ml); only SE from Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Fusobacterium nucleatum caused a significant effect (greater than 90% inhibition or overt morphological abnormalities) in the HGFs as determined by any of the criteria employed. SE from Capnocytophaga sputigena, Eikenella corrodens, or Wolinella recta also inhibited cell proliferation and thymidine incorporation at this dosage; however, the degree of inhibition (5-50%) was consistently, clearly less than that of the first group of three organisms named above. The SE of the three other organisms tested (Actinomyces odontolyticus, Bacteroides intermedius, and Streptococcus sanguis) had little or no effect (0-10% inhibition) at this concentration. The data suggest that the outcome of the interaction between bacterial components and normal resident cells of the periodontium is, at least in part, a function of the bacterial species

  20. Polymicrobial Oral Infection with Four Periodontal Bacteria Orchestrates a Distinct Inflammatory Response and Atherosclerosis in ApoE null Mice.

    Science.gov (United States)

    Chukkapalli, Sasanka S; Velsko, Irina M; Rivera-Kweh, Mercedes F; Zheng, Donghang; Lucas, Alexandra R; Kesavalu, Lakshmyya

    2015-01-01

    Periodontal disease (PD) develops from a synergy of complex subgingival oral microbiome, and is linked to systemic inflammatory atherosclerotic vascular disease (ASVD). To investigate how a polybacterial microbiome infection influences atherosclerotic plaque progression, we infected the oral cavity of ApoE null mice with a polybacterial consortium of 4 well-characterized periodontal pathogens, Porphyromonas gingivalis, Treponema denticola, Tannerealla forsythia and Fusobacterium nucleatum, that have been identified in human atherosclerotic plaque by DNA screening. We assessed periodontal disease characteristics, hematogenous dissemination of bacteria, peripheral T cell response, serum inflammatory cytokines, atherosclerosis risk factors, atherosclerotic plaque development, and alteration of aortic gene expression. Polybacterial infections have established gingival colonization in ApoE null hyperlipidemic mice and displayed invasive characteristics with hematogenous dissemination into cardiovascular tissues such as the heart and aorta. Polybacterial infection induced significantly higher levels of serum risk factors oxidized LDL (p Periodontal microbiome infection is associated with significant decreases in Apoa1, Apob, Birc3, Fga, FgB genes that are associated with atherosclerosis. Periodontal infection for 12 weeks had modified levels of inflammatory molecules, with decreased Fas ligand, IL-13, SDF-1 and increased chemokine RANTES. In contrast, 24 weeks of infection induced new changes in other inflammatory molecules with reduced KC, MCSF, enhancing GM-CSF, IFNγ, IL-1β, IL-13, IL-4, IL-13, lymphotactin, RANTES, and also an increase in select inflammatory molecules. This study demonstrates unique differences in the host immune response to a polybacterial periodontal infection with atherosclerotic lesion progression in a mouse model.

  1. Regulation of Ghrelin Receptor by Periodontal Bacteria In Vitro and In Vivo.

    Science.gov (United States)

    Nokhbehsaim, Marjan; Damanaki, Anna; Nogueira, Andressa Vilas Boas; Eick, Sigrun; Memmert, Svenja; Zhou, Xiaoyan; Nanayakkara, Shanika; Götz, Werner; Cirelli, Joni Augusto; Jäger, Andreas; Deschner, James

    2017-01-01

    Ghrelin plays a major role in obesity-related diseases which have been shown to be associated with periodontitis. This study sought to analyze the expression of the functional receptor for ghrelin (GHS-R1a) in periodontal cells and tissues under microbial conditions in vitro and in vivo . The GHS-R1a expression in human periodontal cells challenged with the periodontopathogen Fusobacterium nucleatum , in gingival biopsies from periodontally healthy and diseased individuals, and from rats with and without ligature-induced periodontitis was analyzed by real-time PCR, immunocytochemistry, and immunofluorescence. F. nucleatum induced an initial upregulation and subsequent downregulation of GHS-R1a in periodontal cells. In rat experimental periodontitis, the GHS-R1a expression at periodontitis sites was increased during the early stage of periodontitis, but significantly reduced afterwards, when compared with healthy sites. In human gingival biopsies, periodontally diseased sites showed a significantly lower GHS-R1a expression than the healthy sites. The expression of the functional ghrelin receptor in periodontal cells and tissues is modulated by periodontal bacteria. Due to the downregulation of the functional ghrelin receptor by long-term exposure to periodontal bacteria, the anti-inflammatory actions of ghrelin may be diminished in chronic periodontal infections, which could lead to an enhanced periodontal inflammation and tissue destruction.

  2. Real-time PCR quantification of six periodontal pathogens in saliva samples from healthy young adults.

    Science.gov (United States)

    Zhou, Xiaodong; Liu, Xiaoli; Li, Jing; Aprecio, Raydolfo M; Zhang, Wu; Li, Yiming

    2015-05-01

    The use of saliva as a diagnostic fluid for the evaluation of periodontal health has gained attention recently. Most published real-time PCR assays focused on quantification of bacteria in subgingival plaque, not in saliva. The aims of this study were to develop a real-time PCR assay for quantification of six periodontal pathogens in saliva and to establish a relationship between the amount of DNA (fg) and colony-forming unit (CFU). TaqMan primers/probe sets were used for the detection of Aggregatibacter actinomycetemcomitans (Aa), Eikenella corrodens (Ec), Fusobacterium nucleatum (Fn), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Tannerella forsythia (Tf), and total bacteria. Six periodontal pathogens and total bacteria in saliva from 24 periodontally healthy individuals were determined. The relationship between the amount of DNA (fg) and CFU was established by measuring the concentrations of extracted bacterial DNA and CFU per milliliter of bacteria on agar plates. Fn, Ec, and Pi were detected in all saliva samples, while 58.5, 45.8, and 33.3% were detected for Tf, Pg, and Aa, respectively. Numbers of Ec and Fn in saliva were highly correlated (R(2) = 0.93, P periodontal pathogens in saliva and estimate the number of live bacteria (CFU). This real-time PCR assay in combination with the relationship between DNA (fg) and CFU has the potential to be an adjunct in evaluation of periodontal health status.

  3. Detection of five potentially periodontal pathogenic bacteria in peri-implant disease: A comparison of PCR and real-time PCR.

    Science.gov (United States)

    Schmalz, Gerhard; Tsigaras, Sandra; Rinke, Sven; Kottmann, Tanja; Haak, Rainer; Ziebolz, Dirk

    2016-07-01

    The aim of this study was to compare the microbial analysis methods of polymerase chain reaction (PCR) and real-time PCR (RT-PCR) in terms of detection of five selected potentially periodontal pathogenic bacteria in peri-implant disease. Therefore 45 samples of healthy, mucositis and peri-implantitis (n = 15 each) were assessed according to presence of the following bacteria using PCR (DNA-strip technology) and RT-PCR (fluorescent dye SYBR green-system): Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tanerella forsythia (Tf), and Fusobacterium nucleatum (Fn). There were no significant correlations between the bacterial and disease patterns, so the benefit of using microbiological tests for the diagnosis of peri-implant diseases is questionable. Correlations between the methods were highest for Tf (Kendall's Tau: 0.65, Spearman: 0.78), Fn (0.49, 0.61) and Td (0.49, 0.59). For Aa (0.38, 0.42) and Pg (0.04, 0.04), lower correlation values were detected. Accordingly, conventional semi-quantitative PCR seems to be sufficient for analyzing potentially periodontal pathogenic bacterial species. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Periodontal bacteria in the genital tract: are they related to adverse pregnancy outcome?

    Science.gov (United States)

    Cassini, M A; Pilloni, A; Condò, S G; Vitali, L A; Pasquantonio, G; Cerroni, L

    2013-01-01

    One of the most important factors implicated in preterm birth (PTB) is acute genitourinary tract infection. The bacteria causing chronic periodontal inflammation include Gram-negative rods and anaerobes similar to those found in women with bacterial vaginosis. The aim of this prospective study is to investigate the relationship between oral and vaginal microflora and preterm low birth weight. Real-time polymerase chain reaction was used to detect both the presence and level of six periodontitis-related species: Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Tannerella forsythia (Tf), Treponema denticola (Td), Fusobacterium nucleatum ssp(Fn), and Prevotella intermedia (Pi) for both oral samples of subgingival plaque and cervical samples, obtained from 80 patients, during gynaecological examinations. The more representative oral pathogen (less than 60 percent) species in oral samples of preterm and term group were Tf, Td, and Fn. 24.4 percent of pregnant women presented periodontal pathogens in vaginal swab; the most representative species with a percentage over 0.1 percent of total bacteria in genital tract of preterm group were Tf, Td, and Piwith a positive correlation (less than 0.5). The presence of the bacterium T. denticolain the vagina, regardless of the amount, adversely affects preterm delivery.

  5. Total antioxidant capacity and total oxidant status in saliva of periodontitis patients in relation to bacterial load

    Directory of Open Access Journals (Sweden)

    Taowen eZhang

    2016-01-01

    Full Text Available The detection of salivary biomarkers has a potential application in early diagnosis and monitoring of periodontal inflammation. However, searching sensitive salivary biomarkers for periodontitis is still ongoing. Oxidative stress is supposed to play an important role in periodontitis progression and tissue destruction. In this cross-sectional study, we investigated total antioxidant capacity (TAC and total oxidant status (TOS in saliva of periodontitis patients compared to healthy controls and their relationship with periodontopathic bacteria and periodontal disease severity. Unstimulated saliva was collected from 45 patients with generalized severe periodontitis and 37 healthy individuals and the TAC/TOS were measured. In addition, salivary levels of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Fusobacterium nucleatum in saliva were measured. Salivary TAC was lower in periodontitis patients compared to healthy controls. Moreover, a significant negative correlation of salivary TAC with clinical attachment loss was observed in periodontitis patients. No significant difference in the salivary TOS was observed between periodontitis patients and healthy controls. Bacterial load was enhanced in periodontitis patients and exhibited correlation with periodontal disease severity but not with salivary TAC/TOS. Our data suggest that changes in antioxidant capacity in periodontitis patients are not associated with increased bacterial load and are probably due to a dysregulated immune response.

  6. 16S rRNA based microarray analysis of ten periodontal bacteria in patients with different forms of periodontitis.

    Science.gov (United States)

    Topcuoglu, Nursen; Kulekci, Guven

    2015-10-01

    DNA microarray analysis is a computer based technology, that a reverse capture, which targets 10 periodontal bacteria (ParoCheck) is available for rapid semi-quantitative determination. The aim of this three-year retrospective study was to display the microarray analysis results for the subgingival biofilm samples taken from patient cases diagnosed with different forms of periodontitis. A total of 84 patients with generalized aggressive periodontitis (GAP,n:29), generalized chronic periodontitis (GCP, n:25), peri-implantitis (PI,n:14), localized aggressive periodontitis (LAP,n:8) and refractory chronic periodontitis (RP,n:8) were consecutively selected from the archives of the Oral Microbiological Diagnostic Laboratory. The subgingival biofilm samples were analyzed by the microarray-based identification of 10 selected species. All the tested species were detected in the samples. The red complex bacteria were the most prevalent with very high levels in all groups. Fusobacterium nucleatum was detected in all samples at high levels. The green and blue complex bacteria were less prevalent compared with red and orange complex, except Aggregatibacter actinomycetemcomitas was detected in all LAP group. Positive correlations were found within all the red complex bacteria and between red and orange complex bacteria especially in GCP and GAP groups. Parocheck enables to monitoring of periodontal pathogens in all forms of periodontal disease and can be alternative to other guiding and reliable microbiologic tests. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Detection of selected periodontal bacteria in preschool children affected by early childhood caries.

    Science.gov (United States)

    Pantuckova, Pavla; Bartosova, Michaela; Broukal, Zdenek; Kukletova, Martina; Holla, Lydie Izakovicova

    2016-11-01

    The aim of this study was to compare the detection frequency of periodontal bacteria in dental plaque in children with early childhood caries (ECC) with and without gingival inflammation. A convenience sample of 25 preschool children (mean age 3.61 years, SD 1.42) was recruited. Dental plaque was taken from periodontal areas with and without visible signs of inflammation and processed using the StomaGene® (Protean s.r.o. Czech Republic) and ParoCheck® 20 (Greiner Bio-one GmbH, Germany) detection kits. The two sample t tests between percents for differences between inflammatory and healthy sites and kappa statistics for the agreement of both systems were used. At the inflammatory sites, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were significantly more frequently detected by StomaGene® while Fusobacterium nucleatum, A. actinomycetemcomitans, Tanarella forsythia and Prevotella intermedia were significantly more frequently identified by ParoCheck® test. The agreement between the two detection systems was substantial for A. actinomycetemcomitans and F. nucleatum in the samples collected from inflamed sites and only for F. nucleatum from clinically healthy sites. Therefore, we recommend that the same system should be used when the same patient is examined repeatedly.

  8. CLINICAL USE OF MODERN FLUOROQUINOLONES IN DENTAL IMPLANTATION AND MAXILLARY SINUS LIFTING

    Directory of Open Access Journals (Sweden)

    S.A. Novikov

    2009-03-01

    Full Text Available New fluoroquinolones like levofloxacine, with broad-spectrum of effect which is also active against anaerobic bacteria, is a prospective drug for the prevention of the inflammatory complications in implant dentistry and maxillary sinus augmentation. 34 patients have been selected for this study, including 16 women and 18 men aged 18-65. All the patients have been classified into the main and control groups. Patiens of the first group have been prescribed 500 mg of levofloxacine before and after the operation two times a day for 10 days. Patients of the control group have been treated with amocxicilline. In the control group, which has used amoxicilline, Streptococcus sanguis, Streptococcus salivarius and anaerobic bacteria have been identified. The amount of Prevotella intermedia is insignificant. The same picture has been found with Fusobacterium spp. The rate of Actinomyces spp. was insignificant over the whole period of study. Patients of the experimental group had a slightly different distribution of bacteria. On the 10th day after the operation in the main patient group no periodontal pathogenesis, which may cause inflammatory complications have been identified. During the early post-operative period no significant differences have been observed among the patients of both groups.

  9. Hydrogen sulfide production from cysteine and homocysteine by periodontal and oral bacteria.

    Science.gov (United States)

    Yoshida, Akihiro; Yoshimura, Mamiko; Ohara, Naoya; Yoshimura, Shigeru; Nagashima, Shiori; Takehara, Tadamichi; Nakayama, Koji

    2009-11-01

    Hydrogen sulfide is one of the predominant volatile sulfur compounds (VSCs) produced by oral bacteria. This study developed and evaluated a system for detecting hydrogen sulfide production by oral bacteria. L-methionine-alpha-deamino-gamma-mercaptomethane-lyase (METase) and beta carbon-sulfur (beta C-S) lyase were used to degrade homocysteine and cysteine, respectively, to produce hydrogen sulfide. Enzymatic reactions resulting in hydrogen sulfide production were assayed by reaction with bismuth trichloride, which forms a black precipitate when mixed with hydrogen sulfide. The enzymatic activities of various oral bacteria that result in hydrogen sulfide production and the capacity of bacteria from periodontal sites to form hydrogen sulfide in reaction mixtures containing L-cysteine or DL-homocysteine were assayed. With L-cysteine as the substrate, Streptococcus anginosus FW73 produced the most hydrogen sulfide, whereas Porphyromonas gingivalis American Type Culture Collection (ATCC) 33277 and W83 and Fusobacterium nucleatum ATCC 10953 produced approximately 35% of the amount produced by the P. gingivalis strains. Finally, the hydrogen sulfide found in subgingival plaque was analyzed. Using bismuth trichloride, the hydrogen sulfide produced by oral bacteria was visually detectable as a black precipitate. Hydrogen sulfide production by oral bacteria was easily analyzed using bismuth trichloride. However, further innovation is required for practical use.

  10. Discrimination of the oral microbiota associated with high hydrogen sulfide and methyl mercaptan production.

    Science.gov (United States)

    Takeshita, Toru; Suzuki, Nao; Nakano, Yoshio; Yasui, Masaki; Yoneda, Masahiro; Shimazaki, Yoshihiro; Hirofuji, Takao; Yamashita, Yoshihisa

    2012-01-01

    Both hydrogen sulfide (H2S) and methyl mercaptan (CH(3)SH) are frequently detected in large amounts in malodorous mouth air. We investigated the bacterial composition of saliva of 30 subjects with severe oral malodor exhibiting extreme CH(3)SH/H(2)S ratios (high H(2)S but low CH(3)SH concentrations, n 5 14; high CH(3)SH but low H2S concentrations, n 5 16) and 13 subjects without malodor, using barcoded pyrosequencing analysis of the 16S rRNA gene. Phylogenetic community analysis with the UniFrac distance metric revealed a distinct bacterial community structure in each malodor group. The H2S group showed higher proportions of the genera Neisseria, Fusobacterium, Porphyromonas and SR1 than the other two groups, whereas the CH(3)SH group had higher proportions of the genera Prevotella, Veillonella,Atopobium, Megasphaera, and Selenomonas. Our results suggested that distinct bacterial populations in the oral microbiota are involved in production of high levels of H2S and CH3SH in the oral cavity.

  11. Biomechanical Loading Modulates Proinflammatory and Bone Resorptive Mediators in Bacterial-Stimulated PDL Cells

    Directory of Open Access Journals (Sweden)

    Andressa Vilas Boas Nogueira

    2014-01-01

    Full Text Available The present study aimed to evaluate in vitro whether biomechanical loading modulates proinflammatory and bone remodeling mediators production by periodontal ligament (PDL cells in the presence of bacterial challenge. Cells were seeded on BioFlex culture plates and exposed to Fusobacterium nucleatum ATCC 25586 and/or cyclic tensile strain (CTS of low (CTSL and high (CTSH magnitudes for 1 and 3 days. Synthesis of cyclooxygenase-2 (COX2 and prostaglandin E2 (PGE2 was evaluated by ELISA. Gene expression and protein secretion of osteoprotegerin (OPG and receptor activator of nuclear factor kappa-B ligand (RANKL were evaluated by quantitative RT-PCR and ELISA, respectively. F. nucleatum increased the production of COX2 and PGE2, which was further increased by CTS. F. nucleatum-induced increase of PGE2 synthesis was significantly (P<0.05 increased when CTSH was applied at 1 and 3 days. In addition, CTSH inhibited the F. nucleatum-induced upregulation of OPG at 1 and 3 days, thereby increasing the RANKL/OPG ratio. OPG and RANKL mRNA results correlated with the protein results. In summary, our findings provide original evidence that CTS can enhance bacterial-induced syntheses of molecules associated with inflammation and bone resorption by PDL cells. Therefore, biomechanical, such as orthodontic or occlusal, loading may enhance the bacterial-induced inflammation and destruction in periodontitis.

  12. FISHing for gutta-percha-adhered biofilms in purulent post-treatment apical periodontitis.

    Science.gov (United States)

    Zehnder, M; Rechenberg, D-K; Thurnheer, T; Lüthi-Schaller, H; Belibasakis, G N

    2017-06-01

    This study investigated the possibility of depicting individual taxa in clinically relevant biofilms using fluorescent in situ hybridization (FISH). Gutta-percha samples were collected from the apical aspect of root canals associated with a chronic apical abscess (test samples, n = 8). Corresponding control samples were obtained from previously filled root canals with apparently normal periapical tissues (n = 3). The transport medium was investigated for detached biofilm fragments using FISH staining and conventional epifluorescence microscopy. Gutta-percha samples were stained by multiplex FISH, and inspected using confocal laser scanning microscopy. FISH of the transport medium confirmed the presence of the main species formerly identified by conventional methods in post-treatment purulent endodontic infections, most prominently Fusobacterium spp., Bacteroidetes and Prevotellaceae. Treponemes were identified in five of eight cases associated with purulent infections, but Enterococcus faecalis and Staphylococcus spp. were not identified. The biofilms on gutta-percha from root canals associated with apical periodontitis showed dense aggregates of variable composition. Control samples contained few, if any, bacteria in the transport medium, and featured no biofilms on the respective gutta-percha specimens. The current study revealed some direct, visual in situ information on the nature of biofilms associated with purulent periapical infections in man. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  13. Review of the gut microbiome and esophageal cancer: Pathogenesis and potential clinical implications.

    Science.gov (United States)

    Baba, Yoshifumi; Iwatsuki, Masaaki; Yoshida, Naoya; Watanabe, Masayuki; Baba, Hideo

    2017-06-01

    Esophageal cancer ranks among the most aggressive malignant diseases. The limited improvements in treatment outcomes provided by conventional therapies have prompted us to seek innovative strategies for treating this cancer. More than 100 trillion microorganisms inhabit the human intestinal tract and play a crucial role in health and disease conditions, including cancer. The human intestinal microbiome is thought to influence tumor development and progression in the gastrointestinal tract by various mechanisms. For example, Fusobacterium nucleatum , which primarily inhabits the oral cavity and causes periodontal disease, might contribute to aggressive tumor behavior through activation of chemokines such as CCL20 in esophageal cancer tissue. Composition of the intestinal microbiota is influenced by diet, lifestyle, antibiotics, and pro- and prebiotics. Therefore, by better understanding how the bacterial microbiota contributes to esophageal carcinogenesis, we might develop novel cancer prevention and treatment strategies through targeting the gastrointestinal microflora. This review discusses the current knowledge, available data and information on the relationship of microbiota with esophagitis, Barrett's esophagus, esophageal adenocarcinoma and squamous cell carcinoma.

  14. Antianaerobic Antimicrobials: Spectrum and Susceptibility Testing

    Science.gov (United States)

    Wexler, Hannah M.; Goldstein, Ellie J. C.

    2013-01-01

    SUMMARY Susceptibility testing of anaerobic bacteria recovered from selected cases can influence the choice of antimicrobial therapy. The Clinical and Laboratory Standards Institute (CLSI) has standardized many laboratory procedures, including anaerobic susceptibility testing (AST), and has published documents for AST. The standardization of testing methods by the CLSI allows comparisons of resistance trends among various laboratories. Susceptibility testing should be performed on organisms recovered from sterile body sites, those that are isolated in pure culture, or those that are clinically important and have variable or unique susceptibility patterns. Organisms that should be considered for individual isolate testing include highly virulent pathogens for which susceptibility cannot be predicted, such as Bacteroides, Prevotella, Fusobacterium, and Clostridium spp.; Bilophila wadsworthia; and Sutterella wadsworthensis. This review describes the current methods for AST in research and reference laboratories. These methods include the use of agar dilution, broth microdilution, Etest, and the spiral gradient endpoint system. The antimicrobials potentially effective against anaerobic bacteria include beta-lactams, combinations of beta-lactams and beta-lactamase inhibitors, metronidazole, chloramphenicol, clindamycin, macrolides, tetracyclines, and fluoroquinolones. The spectrum of efficacy, antimicrobial resistance mechanisms, and resistance patterns against these agents are described. PMID:23824372

  15. Antibiotic susceptibility profiles of anaerobic pathogens in The Netherlands.

    Science.gov (United States)

    Veloo, A C M; van Winkelhoff, A J

    2015-02-01

    The antibiotic susceptibility profile of the Bacteroides fragilis group, Gram-positive anaerobic cocci (GPAC), Fusobacterium spp., Prevotella spp., Veillonella spp. and Bilophila wadsworthia for amoxicillin, amoxicillin-clavulanic acid, clindamycin and metronidazole was determined. Human clinical isolates were isolated between 2011 and 2013 at the Microbiological Diagnostic Laboratory of the University Medical Center Groningen, The Netherlands and subjected to MALDI-TOF MS identification and susceptibility testing using E-test for MIC determination. Differences in clindamycin susceptibility between species of the B. fragilis group and GPAC were observed, with Bacteroides ovatus and Peptoniphilus harei having the highest resistance rates. Compared to other European countries, in The Netherlands the MIC90 for clindamycin of fusobacteria is low. Metronidazole resistance was first encountered in the genus Prevotella in 2013, but not in species of GPAC as reported in Belgium and Bulgaria. The differences in clindamycin resistance between the different European countries and reports of metronidazole resistance within the genera Prevotella and GPAC warrant more extensive susceptibility studies on anaerobic pathogens. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. The Biofilm Community-Rebels with a Cause.

    Science.gov (United States)

    Aruni, A Wilson; Dou, Yuetan; Mishra, Arunima; Fletcher, Hansel M

    2015-03-01

    Oral Biofilms are one of the most complex and diverse ecosystem developed by successive colonization of more than 600 bacterial taxa. Development starts with the attachment of early colonizers such as Actinomyces species and oral streptococci on the acquired pellicle and tooth enamel. These bacteria not only adhere to tooth surface but also interact with each other and lay foundation for attachment of bridging colonizer such as Fusobacterium nucleatum followed by late colonizers including the red complex species: Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola -the founders of periodontal disease. As the biofilm progresses from supragingival sites to subgingival sites, the environment changes from aerobic to anaerobic thus favoring the growth of mainly Gram-negative obligate anaerobes while restricting the growth of the early Gram-positive facultative aerobes. Microbes present at supragingival level are mainly related to gingivitis and root-caries whereas subgingival species advance the destruction of teeth supporting tissues and thus causing periodontitis. This review summarizes our present understanding and recent developments on the characteristic features of supra- and subgingival biofilms, interaction between different genera and species of bacteria constituting these biofilms and draws our attention to the role of some of the recently discovered members of the oral community.

  17. Intake of Meat Proteins Substantially Increased the Relative Abundance of Genus Lactobacillus in Rat Feces.

    Directory of Open Access Journals (Sweden)

    Yingying Zhu

    Full Text Available Diet has been shown to have a critical influence on gut bacteria and host health, and high levels of red meat in diet have been shown to increase colonic DNA damage and thus be harmful to gut health. However, previous studies focused more on the effects of meat than of meat proteins. In order to investigate whether intake of meat proteins affects the composition and metabolic activities of gut microbiota, feces were collected from growing rats that were fed with either meat proteins (from beef, pork or fish or non-meat proteins (casein or soy for 14 days. The resulting composition of gut microbiota was profiled by sequencing the V4-V5 region of the 16S ribosomal RNA genes and the short chain fatty acids (SCFAs were analyzed using gas chromatography. The composition of gut microbiota and SCFA levels were significantly different between the five diet groups. At a recommended dose of 20% protein in the diet, meat protein-fed rats had a higher relative abundance of the beneficial genus Lactobacillus, but lower levels of SCFAs and SCFA-producing bacteria including Fusobacterium, Bacteroides and Prevotella, compared with the soy protein-fed group. Further work is needed on the regulatory pathways linking dietary protein intake to gut microbiota.

  18. Microbiology and Treatment of Acute Apical Abscesses

    Science.gov (United States)

    Rôças, Isabela N.

    2013-01-01

    SUMMARY Acute apical abscess is the most common form of dental abscess and is caused by infection of the root canal of the tooth. It is usually localized intraorally, but in some cases the apical abscess may spread and result in severe complications or even mortality. The reasons why dental root canal infections can become symptomatic and evolve to severe spreading and sometimes life-threatening abscesses remain elusive. Studies using culture and advanced molecular microbiology methods for microbial identification in apical abscesses have demonstrated a multispecies community conspicuously dominated by anaerobic bacteria. Species/phylotypes commonly found in these infections belong to the genera Fusobacterium, Parvimonas, Prevotella, Porphyromonas, Dialister, Streptococcus, and Treponema. Advances in DNA sequencing technologies and computational biology have substantially enhanced the knowledge of the microbiota associated with acute apical abscesses and shed some light on the etiopathogeny of this disease. Species richness and abundance and the resulting network of interactions among community members may affect the collective pathogenicity and contribute to the development of acute infections. Disease modifiers, including transient or permanent host-related factors, may also influence the development and severity of acute abscesses. This review focuses on the current evidence about the etiology and treatment of acute apical abscesses and how the process is influenced by host-related factors and proposes future directions in research, diagnosis, and therapeutic approaches to deal with this disease. PMID:23554416

  19. Coenzyme Recognition and Gene Regulation by a Flavin Mononucleotide Riboswitch

    Energy Technology Data Exchange (ETDEWEB)

    Serganov, A.; Huang, L; Patel, D

    2009-01-01

    The biosynthesis of several protein cofactors is subject to feedback regulation by riboswitches. Flavin mononucleotide (FMN)-specific riboswitches also known as RFN elements, direct expression of bacterial genes involved in the biosynthesis and transport of riboflavin (vitamin B2) and related compounds. Here we present the crystal structures of the Fusobacterium nucleatum riboswitch bound to FMN, riboflavin and antibiotic roseoflavin. The FMN riboswitch structure, centred on an FMN-bound six-stem junction, does not fold by collinear stacking of adjacent helices, typical for folding of large RNAs. Rather, it adopts a butterfly-like scaffold, stapled together by opposingly directed but nearly identically folded peripheral domains. FMN is positioned asymmetrically within the junctional site and is specifically bound to RNA through interactions with the isoalloxazine ring chromophore and direct and Mg{sup 2+}-mediated contacts with the phosphate moiety. Our structural data, complemented by binding and footprinting experiments, imply a largely pre-folded tertiary RNA architecture and FMN recognition mediated by conformational transitions within the junctional binding pocket. The inherent plasticity of the FMN-binding pocket and the availability of large openings make the riboswitch an attractive target for structure-based design of FMN-like antimicrobial compounds. Our studies also explain the effects of spontaneous and antibiotic-induced deregulatory mutations and provided molecular insights into FMN-based control of gene expression in normal and riboflavin-overproducing bacterial strains.

  20. Novel model for multispecies biofilms that uses rigid gas-permeable lenses.

    Science.gov (United States)

    Peyyala, Rebecca; Kirakodu, Sreenatha S; Ebersole, Jeffrey L; Novak, Karen F

    2011-05-01

    Oral biofilms comprise complex multispecies consortia aided by specific inter- and intraspecies interactions occurring among commensals and pathogenic bacterial species. Oral biofilms are primary initiating factors of periodontal disease, although complex multifactorial biological influences, including host cell responses, contribute to the individual outcome of the disease. To provide a system to study initial stages of interaction between oral biofilms and the host cells that contribute to the disease process, we developed a novel in vitro model system to grow biofilms on rigid gas-permeable contact lenses (RGPLs), which enable oxygen to permeate through the lens material. Bacterial species belonging to early- and late-colonizing groups were successfully established as single- or three-species biofilms, with each group comprising Streptococcus gordonii, Streptococcus oralis, and Streptococcus sanguinis; S. gordonii, Actinomyces naeslundii, and Fusobacterium nucleatum; or S. gordonii, F. nucleatum, and Porphyromonas gingivalis. Quantification of biofilm numbers by quantitative PCR (qPCR) revealed substantial differences in the magnitude of bacterial numbers in single-species and multispecies biofilms. We evaluated cell-permeable conventional nucleic acid stains acridine orange, hexidium iodide, and Hoechst 33258 and novel SYTO red, blue, and green fluorochromes for their effect on bacterial viability and fluorescence yield to allow visualization of the aggregates of individual bacterial species by confocal laser scanning microscopy (CLSM). Substantial differences in the quantity and distribution of the species in the multispecies biofilms were identified. The specific features of these biofilms may help us better understand the role of various bacteria in local challenge of oral tissues.

  1. Total Antioxidant Capacity and Total Oxidant Status in Saliva of Periodontitis Patients in Relation to Bacterial Load

    Science.gov (United States)

    Zhang, Taowen; Andrukhov, Oleh; Haririan, Hady; Müller-Kern, Michael; Liu, Shutai; Liu, Zhonghao; Rausch-Fan, Xiaohui

    2016-01-01

    The detection of salivary biomarkers has a potential application in early diagnosis and monitoring of periodontal inflammation. However, searching sensitive salivary biomarkers for periodontitis is still ongoing. Oxidative stress is supposed to play an important role in periodontitis progression and tissue destruction. In this cross-sectional study, we investigated total antioxidant capacity (TAC) and total oxidant status (TOS) in saliva of periodontitis patients compared to healthy controls and their relationship with periodontopathic bacteria and periodontal disease severity. Unstimulated saliva was collected from 45 patients with generalized severe periodontitis and 37 healthy individuals and the TAC/TOS were measured. In addition, salivary levels of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, and Fusobacterium nucleatum in saliva were measured. Salivary TAC was lower in periodontitis patients compared to healthy controls. Moreover, a significant negative correlation of salivary TAC with clinical attachment loss was observed in periodontitis patients. No significant difference in the salivary TOS was observed between periodontitis patients and healthy controls. Bacterial load was enhanced in periodontitis patients and exhibited correlation with periodontal disease severity but not with salivary TAC/TOS. Our data suggest that changes in antioxidant capacity in periodontitis patients are not associated with increased bacterial load and are probably due to a dysregulated immune response. PMID:26779448

  2. The Kampo medicine Rokumigan possesses antibiofilm, anti-inflammatory, and wound healing properties.

    Science.gov (United States)

    Liao, James; Azelmat, Jabrane; Zhao, Lei; Yoshioka, Masami; Hinode, Daisuke; Grenier, Daniel

    2014-01-01

    Periodontal diseases, which are inflammatory diseases of bacterial origin affecting the tooth-supporting tissues, are characterized by inflammation and destruction of gingival connective tissue and alveolar bone, and may lead to tooth loss. The aim of the study was to investigate Rokumigan, a Kampo Japanese traditional medicine made of six different plants, for its capacity to prevent biofilm formation by Fusobacterium nucleatum, to inhibit interleukin-6 (IL-6) and interleukin-8 (IL-8) secretion by mucosal cells, and to promote wound healing in a fibroblast model. Using a microplate colorimetric assay, Rokumigan prevented biofilm formation by F. nucleatum, while it had no effect on bacterial growth. Rokumigan inhibited IL-6 secretion in both epithelial cells and fibroblasts stimulated with lipopolysaccharide. However, it caused no significant inhibition of IL-8 secretion by both cell types. Rokumigan significantly increased proliferation and migration of gingival fibroblasts in a wound healing assay. In conclusion, the Kampo formulation Rokumigan, through suppression of biofilm formation by F. nucleatum, inhibition of IL-6 secretion by gingival epithelial cells and fibroblasts, and promotion of wound healing in a fibroblast model, may have potential application for periodontal diseases.

  3. Experimental alveolitis in rats: microbiological, acute phase response and histometric characterization of delayed alveolar healing.

    Science.gov (United States)

    Rodrigues, Moacyr Tadeu Vicente; Cardoso, Camila Lopes; Carvalho, Paulo Sérgio Perri de; Cestari, Tânia Mary; Feres, Magda; Garlet, Gustavo Pompermaier; Ferreira, Osny

    2011-01-01

    The pathogenesis of alveolitis is not well known and therefore experimental situations that mimic some features of this disease should be developed. In this study, the evolution of the experimentally induced infection in rat sockets is characterized, which leads to clinical signs of suppurative alveolitis with remarkable wound healing disturbs. Non-infected (Group I) and experimentally infected sockets in Rattus novergicus (Group II) were histometrically evaluated regarding the kinetics of alveolar healing. In addition, the characterization of the present bacteria in inoculation material and the serum levels of C-reactive protein (CRP) were performed. The detected species were Capnocytophaga ochracea, Fusobacterium nucleatum ss nucleatum, Prevotella melaninogenica, Streptococcus anginosus, Treponema socranskii and Streptococcus sanguis. All experimentally infected rats developed suppurative alveolitis, showing higher levels of CRP in comparison to those non-infected ones. Furthermore, infected rats presented a significant delayed wound healing as measured by the histometric analysis (higher persistent polymorphonuclear infiltrate and lower density of newly formed bone). These findings indicate that rat sockets with experimentally induced infection produced higher levels of serum CRP, showing the potential of disseminated infection and a disturb in the alveolar repair process in an interesting experimental model for alveolitis studies.

  4. Laser Supported Reduction of Specific Microorganisms in the Periodontal Pocket with the Aid of an Er,Cr:YSGG Laser: A Pilot Study

    Directory of Open Access Journals (Sweden)

    N. Gutknecht

    2015-01-01

    Full Text Available Objective. The aim of this study was to evaluate the effectiveness of a radial firing tip of an Er,Cr:YSGG laser as an adjunct to a nonsurgical periodontal treatment. Methods. Twelve patients with chronic or aggressive periodontitis were treated by conventional periodontal treatment using ultrasonic devices and hand instruments and, additionally, in two quadrants with an Er,Cr:YSGG laser. A new radial firing tip (RFPT 14-5, Biolase was used with 1.5 W, 30 Hz, 11% air, 20% water, and pulse duration 140 μs. Microbiological smears were taken before treatment, one day after lasing, and three and six months after lasing. Pocket depths of all periodontal sites were measured before and six months after treatment. Results. The total bacterial load of Prevotella intermedia, Tannerella forsythia, Treponema denticola, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans inside the pocket was reduced significantly throughout the whole examination time. Greater pocket depth reductions were observed in all groups. There was a slight higher reduction of pocket depth in the lased group after six months. Conclusions. These results support the thesis that Er,Cr:YSGG laser supported periodontal treatment leads to a significant reduction of periopathogenes and thereby helps the maintenance of periodontal health.

  5. The in Vitro Antimicrobial Efficacy of PDT against Periodontopathogenic Bacteria

    Directory of Open Access Journals (Sweden)

    Philippe A. Haag

    2015-11-01

    Full Text Available Periodontitis, an inflammatory disease, is caused by biofilms with a mixed microbial etiology and involves the progressive destruction of the tooth-supporting tissues. A rising number of studies investigate the clinical potential of photodynamic therapy (PDT as an adjunct during active therapy. The aim of the present review was to evaluate the available literature for the in vitro antimicrobial efficacy of photodynamic therapy focusing on the periodontopathogenic bacteria Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Fusobacterium nucleatum. The focused question was: “Is it possible to decrease (at least 3 log steps or 99.9% or even eliminate bacterial growth by photodynamic therapy in vitro when compared to untreated control groups or control groups treated by placebo?” In general, PDT resulted in a substantial reduction of surviving bacteria. However, not all studies showed the desired reduction or elimination. The ranges of log10-reduction were 0.38 (58% to a complete eradication (100% for P. gingivalis, 0.21 (39% to 100% for A. actinomycetemcomitans and 0.3 (50% to 100% for F. nucleatum. In conclusion, further and particularly more comparable studies are needed to evaluate if PDT can be clinically successful as an adjuvant in periodontal therapy.

  6. Variation of perimplant biofilm induced by non surgical periodontal therapy and the use of probiotics

    Directory of Open Access Journals (Sweden)

    Marcello Gatti

    2010-12-01

    Full Text Available Thanks to improved surgical tecniques the use of dental implants has increased greatly. However, high rates of osseointegrated correctly implants, over the years are undermined by disease of bacterial etiology in the perimplant zone, especially by Gram negative anaerobes such as in gingivitis and periodontitis, in particular: Fusobacterium spp.(F., Treponema denticola (T.d., Tannerella forsythensis (T.f., Aggregatibacter actinomycetemcomitans (A.a., Prevotella intermedia (P.i. e Porphyromonas gingivalis (P.g.. The mechanic treatment (MS results in a reduction of the total bacterial count (TBC and a slight change in the subgingival bacterial microflora towards the less pathogenic species and more like those of a healthy periodontium.Also the use of a probiotic in the form of buccal tablets of Lactobacillus reuteri (L.r., as demonstrated in this study, is thought to improve and modulate the composition of plaque, as it is able to exert an inhibitory effect on oral bacteria that support caries, gingivitis, periodontal and perimplant disease with a combination of different mechanisms.

  7. Antibiofilm and Anti-Inflammatory Activities of Houttuynia cordata Decoction for Oral Care

    Directory of Open Access Journals (Sweden)

    Yasuko Sekita

    2017-01-01

    Full Text Available Dental biofilms that form in the oral cavity play a critical role in the pathogenesis of several infectious oral diseases, including dental caries, periodontal disease, and oral candidiasis. Houttuynia cordata (HC, Saururaceae is a widely used traditional medicine, for both internal and external application. A decoction of dried HC leaves (dHC has long been consumed as a health-promoting herbal tea in Japan. We have recently reported that a water solution of HC poultice ethanol extract (wHCP exerts antimicrobial and antibiofilm effects against several important oral pathogens. It also exhibits anti-inflammatory effects on human keratinocytes. In our current study, we examined the effects of dHC on infectious oral pathogens and inflammation. Our results demonstrated that dHC exerts moderate antimicrobial effects against methicillin-resistant Staphylococcus aureus (MRSA and other oral microorganisms. dHC also exhibited antibiofilm effects against MRSA, Fusobacterium nucleatum (involved in dental plaque formation, and Candida albicans and inhibitory effects on interleukin-8, CCL20, IP-10, and GROα productions by human oral keratinocytes stimulated by Porphyromonas gingivalis lipopolysaccharide (a cause of periodontal disease, without cytotoxic effects. This suggests that dHC exhibits multiple activities in microorganisms and host cells. dHC can be easily prepared and may be effective in preventing infectious oral diseases.

  8. Detection and enumeration of periodontopathogenic bacteria in subgingival biofilm of pregnant women

    Directory of Open Access Journals (Sweden)

    Fernanda Campos Machado

    2012-10-01

    Full Text Available The aim of this study was to use the fluorescence in situ hybridization (FISH technique to test the hypothesis of qualitative and quantitative differences of 8 periodontopathogens between pregnant and non-pregnant women. This cross-sectional study included 20 pregnant women in their second trimester of pregnancy and 20 non-pregnant women. Probing depth, bleeding on probing, clinical attachment level, and presence of calculus were recorded. Subgingival plaque samples were collected and the FISH technique identified the presence and numbers of Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Campylobacter rectus, Porphyromonas gingivalis, Treponema denticola, Fusobacterium nucleatum, Prevotella intermedia and Prevotella nigrescens. The Mann-Whitney U-test was applied to compare the data between the two groups. The mean age, ethnicity, marital status, education, and economic level in both groups were similar. The clinical parameters showed no significant differences between pregnant and non-pregnant women. The numbers of subgingival periodontopathogens were not found to be significantly different between groups, despite the higher mean counts of P. intermedia in pregnant women. Colonization patterns of the different bacteria most commonly associated with periodontal disease were not different in the subgingival plaque of pregnant and non-pregnant women.

  9. Periodontal disease, atherosclerosis, adverse pregnancy outcomes, and head-and-neck cancer.

    Science.gov (United States)

    Han, Y W; Houcken, W; Loos, B G; Schenkein, H A; Tezal, M

    2014-05-01

    Interrelationships between periodontal infection and systemic conditions such as cardiovascular disease, adverse pregnancy outcomes, and head-and-neck cancer have become increasingly appreciated in recent years. Periodontitis is associated with cardiovascular disease (CVD) and, experimentally, with measures of atherosclerosis and endothelial dysfunction. Periodontal therapy may reduce atherosclerotic changes and improve endothelial function. Preliminary findings suggest a role for the genetic locus ANRIL in the pathobiology of both CVD and periodontitis. Periodontal pathogens induce anticardiolipin in periodontitis patients by molecular mimicry of the serum protein β-2 glycoprotein I. These antibodies have biological and pathological activities consistent with those reported for other infection-induced antiphospholipid antibodies. Anticardiolipin may explain some of the observed associations between periodontitis and systemic conditions such as CVD and adverse pregnancy outcomes. The oral commensal Fusobacterium nucleatum (Fn) becomes pathogenic on migration to extra-oral sites. Fn infection of the fetal-placental unit has been linked to pregnancy complications, including preterm birth, stillbirth, and early-onset neonatal sepsis. Reagents aimed at inhibiting or resolving inflammatory responses may be used to treat or prevent pregnancy complications due to bacterial infection. Chronic periodontitis may be independently associated with head-and-neck squamous cell carcinoma (HNSCC) through direct toxic effects of bacteria and their products, and/or through indirect effects of inflammation. Additionally, chronic periodontitis may facilitate the acquisition and persistence of oral HPV infection, a recently emerged risk factor for HNSCC.

  10. Simultaneous detection of periodontal pathogens in subgingival plaque and placenta of women with hypertension in pregnancy.

    Science.gov (United States)

    Swati, P; Thomas, Betsy; Vahab, Saadi Abdul; Kapaettu, Satyamoorthy; Kushtagi, Pralhad

    2012-03-01

    There are many studies documenting increased prevalence of periodontal infection in women with preeclampsia. But, very few studies have attempted to establish causal relationship between the two. To find out causal circumstantial evidence by isolating specific periodontal pathogens in oral and placental samples. Antenatal periodontal screening and subgingival plaque collection was carried out in ten women with hypertension in pregnancy and ten normotensive controls on their hospital admission at term for cesarean delivery. Placental biopsy was obtained after aseptic placental collection at the time of elective cesarean delivery. Subgingival plaque and placental biopsy were studied for Porphyromonas gingivalis, Fusobacterium nucleatum, Treponema denticola, Prevotella intermedia and Aggregatibacter actinomycetemcomitans using quantitative polymerase chain reaction technique. Periodontist and laboratory personnel were unaware of case or control status. Periodontal status was not informed to the obstetrician recruiting the cases and laboratory. Microbiology report was not revealed till end of the study. Periodontal pathogens were found to be high in the group with hypertension than the controls. P gingivalis was found in all the samples from subgingival plaque and placenta, irrespective of the periodontal disease status. In cases with hypertension, periodontal pathogens are present in higher proportion in subgingival plaque and placenta.

  11. Evaluation of periodontal pathogens in amniotic fluid and the role of periodontal disease in pre-term birth and low birth weight.

    Science.gov (United States)

    Ercan, Esra; Eratalay, Kenan; Deren, Ozgur; Gur, Deniz; Ozyuncu, Ozgur; Altun, Belgin; Kanli, Ceyda; Ozdemir, Pınar; Akincibay, Hakan

    2013-01-01

    Pre-term birth and/or low birth weight (PTLBW) is a serious problem in developing countries. The absence of known risk factors in ≈ 50% of PTLBW cases has resulted in a continued search for other causes. The aim of this study was to examine the effect of periodontitis on pregnancy outcomes. Samples were taken from 50 pregnant women who underwent amniocentesis. Polymerase chain reaction was performed on amniotic fluid samples obtained during amniocentesis and on subgingival plaque samples to determine the presence of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia, Campylobacter rectus and Eikenella corrodens. Plaque index, gingival index, bleeding on probing, probing depth and clinical attachment level were evaluated. Medical records were obtained after birth. Social and demographic variables were similar among the Gingivitis (G), Localized Periodontitis (LP) and Generalized Periodontitis (GP) groups. Four subjects gave birth to PTLBW neonates. Campylobacter rectus, T. forsythia, P. gingivalis and F. nucleatum were detected in the amniotic fluid and subgingival plaque samples of three patients who gave birth to PTLBW neonates. The amniotic fluid sample from the fourth patient was not positive for any of the tested pathogens. These findings suggest that the transmission of some periodontal pathogens from the oral cavity of the mother may cause adverse pregnancy outcomes. The results contribute to an understanding of the association between periodontal disease and PTLBW, but further studies are required to better clarify the possible relationship.

  12. Dental plaque development on a hydroxyapatite disk in young adults observed by using a barcoded pyrosequencing approach.

    Science.gov (United States)

    Takeshita, Toru; Yasui, Masaki; Shibata, Yukie; Furuta, Michiko; Saeki, Yoji; Eshima, Nobuoki; Yamashita, Yoshihisa

    2015-01-30

    Dental plaque is a dynamic microbial biofilm ecosystem that comprises hundreds of species including difficult-to-cultivate bacteria. We observed the assembly of a plaque bacterial community through 16S rRNA gene analysis. Plaque samples that accumulated on a hydroxyapatite disk for 1, 2, 3, 4, 5, and 7 days with saliva on day 0 were collected from 19 young adults using a removable resin splint. Quantitative PCR analysis showed that the total bacterial amount gradually increased and reached a plateau on day 4. Barcoded pyrosequencing analysis revealed that the microbial richness and diversity particularly increased between days 5 and 7. A principal coordinate analysis plot based on unweighted UniFrac showed the community assembly in a time-related manner, which became increasingly similar to the salivary microbiota. Facultative anaerobic bacteria such as Streptococcus, Neisseria, Abiotrophia, Gemella, and Rothia were predominant in the plaque bacterial community in the earlier days, whereas obligate anaerobes, such as Porphyromonas, Fusobacterium, Prevotella, and Capnocytophaga showed increased dominance on later days. UniFrac analysis also demonstrated that dental caries experience had a significant effect on the assembly process. Our results reveal the development pattern of the plaque bacterial community as well as the inter-individual differences associated with dental caries experience.

  13. Biofilm-mediated Antibiotic-resistant Oral Bacterial Infections: Mechanism and Combat Strategies.

    Science.gov (United States)

    Kanwar, Indulata; Sah, Abhishek K; Suresh, Preeti K

    2017-01-01

    Oral diseases like dental caries and periodontal disease are directly associated with the capability of bacteria to form biofilm. Periodontal diseases have been associated to anaerobic Gram-negative bacteria forming a subgingival plaque (Porphyromonas gingivalis, Actinobacillus, Prevotella and Fusobacterium). Biofilm is a complex bacterial community that is highly resistant to antibiotics and human immunity. Biofilm communities are the causative agents of biological developments such as dental caries, periodontitis, peri-implantitis and causing periodontal tissue breakdown. The review recapitulates the latest advancements in treatment of clinical biofilm infections and scientific investigations, while these novel anti-biofilm strategies are still in nascent phases of development, efforts dedicated to these technologies could ultimately lead to anti-biofilm therapies that are superior to the current antibiotic treatment. This paper provides a review of the literature focusing on the studies on biofilm in the oral cavity, formation of dental plaque biofilm, drug resistance of bacterial biofilm and the antibiofilm approaches as biofilm preventive agents in dentistry, and their mechanism of biofilm inhibition. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  14. Advanced Caries Microbiota in Teeth with Irreversible Pulpitis.

    Science.gov (United States)

    Rôças, Isabela N; Lima, Kenio C; Assunção, Isauremi V; Gomes, Patrícia N; Bracks, Igor V; Siqueira, José F

    2015-09-01

    Bacterial taxa in the forefront of caries biofilms are candidate pathogens for irreversible pulpitis and are possibly the first ones to invade the pulp and initiate endodontic infection. This study examined the microbiota of the most advanced layers of dentinal caries in teeth with irreversible pulpitis. DNA extracted from samples taken from deep dentinal caries associated with pulp exposures was analyzed for the presence and relative levels of 33 oral bacterial taxa by using reverse-capture checkerboard hybridization assay. Quantification of total bacteria, streptococci, and lactobacilli was also performed by using real-time quantitative polymerase chain reaction. Associations between the target bacterial taxa and clinical signs/symptoms were also evaluated. The most frequently detected taxa in the checkerboard assay were Atopobium genomospecies C1 (53%), Pseudoramibacter alactolyticus (37%), Streptococcus species (33%), Streptococcus mutans (33%), Parvimonas micra (13%), Fusobacterium nucleatum (13%), and Veillonella species (13%). Streptococcus species, Dialister invisus, and P. micra were significantly associated with throbbing pain, S. mutans with pain to percussion, and Lactobacillus with continuous pain (P pulpitis is suspected. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  15. Role of oral microbiome on oral cancers, a review.

    Science.gov (United States)

    Gholizadeh, Pourya; Eslami, Hosein; Yousefi, Mehdi; Asgharzadeh, Mohammad; Aghazadeh, Mohammad; Kafil, Hossein Samadi

    2016-12-01

    The oral cavity is inhibited by many of the bacterial species. Some of them have a key role in the development of oral disease. Interrelationships between oral microbiome and systemic conditions such as head-and-neck cancer have become increasingly appreciated in recent years. Emerging evidence also suggests a link between periodontal disease and oral cancer, and the explanation being that chronic inflammation could be a major factor in both diseases. Squamous cell carcinoma is that the most frequently occurring malignancy of the oral cavity and adjacent sites, representing over 90% of all cancers. The incidence of oral cancer is increasing, significantly among young people and women. Worldwide there are 350,000-400,000 new cases diagnosed every year. Bacteria, viruses, and fungi are strongly implicated as etiological factors in certain cancers. In this review we will discuss the association between the development of oral cancer in potentially malignant oral lesions with chronic periodontitis, chronic Porphyromonas gingivalis, Fusobacterium nucleatum, candida, other microbes and described mechanisms which may be involved in these carcinoma. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  16. Maternal periodontal disease and preeclampsia in Jaipur population

    Directory of Open Access Journals (Sweden)

    Girija Jaiman

    2018-01-01

    Full Text Available Background: Preeclampsia is identified as an important cause for mother and newborn mortality. Inspite of extensive research, the exact etiological relations have not been established. Hence, an attempt has been made in this study to evaluate the relationship between the preeclampsia and maternal periodontal disease. Materials and Methods: The case–control study comprised of thirty pregnant women distributed equally in the case (preeclampsia and control (healthy group. Gingival index, plaque index, bleeding on probing, clinical probing depth, and clinical attachment level were measured in both groups. Microbiologic examination for identification of one red complex organism Porphyromonas gingivalis and one orange complex organism Fusobacterium nucleatum were done in plaque and placental blood of cases and controls. The clinical examinations and collection of placental blood were done 24 h before delivery. Results: Periodontal condition in the preeclamptic women was statistically worse compared with the normotensive women. There was no statistically significant association between microorganisms in plaque and placental blood between normotensive control and preeclamptic pregnant women. The preeclamptic women had significantly higher chances of having newborns weighing <2.5 kg than the normotensive women. Conclusion: The preeclamptic women were associated with significantly higher periodontitis and lower fetal birth weight than normotensive women.

  17. Maternal periodontal disease and preeclampsia in Jaipur population.

    Science.gov (United States)

    Jaiman, Girija; Nayak, Prathibha Anand; Sharma, Sanu; Nagpal, Kiran

    2018-01-01

    Preeclampsia is identified as an important cause for mother and newborn mortality. Inspite of extensive research, the exact etiological relations have not been established. Hence, an attempt has been made in this study to evaluate the relationship between the preeclampsia and maternal periodontal disease. The case-control study comprised of thirty pregnant women distributed equally in the case (preeclampsia) and control (healthy) group. Gingival index, plaque index, bleeding on probing, clinical probing depth, and clinical attachment level were measured in both groups. Microbiologic examination for identification of one red complex organism Porphyromonas gingivalis and one orange complex organism Fusobacterium nucleatum were done in plaque and placental blood of cases and controls. The clinical examinations and collection of placental blood were done 24 h before delivery. Periodontal condition in the preeclamptic women was statistically worse compared with the normotensive women. There was no statistically significant association between microorganisms in plaque and placental blood between normotensive control and preeclamptic pregnant women. The preeclamptic women had significantly higher chances of having newborns weighing <2.5 kg than the normotensive women. The preeclamptic women were associated with significantly higher periodontitis and lower fetal birth weight than normotensive women.

  18. Epizootic rabbit enteropathy inoculum (TEC4): antibiograms and antibiotic fractionation.

    Science.gov (United States)

    Huybens, Nathalie; Houeix, Julien; Licois, Dominique; Mainil, Jacques; Marlier, Didier

    2011-01-01

    Epizootic rabbit enteropathy (ERE) emerged and spread in Europe within the last 13 years causing major economical loss. The aims of the study was to evaluate antibiograms of TEC4, an inoculum composed of an extract of intestinal content of affected rabbits, and to test the potential of different antibiotic-based TEC4 fractions to reproduce the disease. Twenty nine different antibiotic discs were incubated for determining bacteria resistance. In a complementary study, nine tubes of liquid medium were inoculated with TEC4, incubated and added individually with amoxicillin/clavulanic acid, bacitracin, ceftiofur, doxycycline, novobiocin, streptomycyin, tylosin, vancomycin and 0.9% saline solution as control. The content of each tube was washed by centrifugation and suspended in saline. The three most effective antibiotics are florfenicol, amoxycillin/clavulanic acid and tylosin. A high concentration of Clostridium sordelli and Bacillus firmus were isolated in all fractions. Species never cultured from TEC4 were identified as Fusobacterium necrogenes (in vancomycin fraction), Cellulomonas sp (in novobiocin fraction) and Bacteroides distasonis (in doxycycline fraction). The ERE was reproduced when bacitracin, doxycycline and 0.9% fractions were inoculated. Rabbits showed ERE clinical signs with the specific drop in daily weight gain.

  19. Emerging role of bacteria in oral carcinogenesis: a review with special reference to perio-pathogenic bacteria.

    Science.gov (United States)

    Perera, Manosha; Al-Hebshi, Nezar Noor; Speicher, David J; Perera, Irosha; Johnson, Newell W

    2016-01-01

    Oral cancer, primarily oral squamous cell carcinoma (OSCC), continues to be a major global health problem with high incidence and low survival rates. While the major risk factors for this malignancy, mostly lifestyle related, have been identified, around 15% of oral cancer cases remain unexplained. In light of evidence implicating bacteria in the aetiology of some cancer types, several epidemiological studies have been conducted in the last decade, employing methodologies ranging from traditional culture techniques to 16S rRNA metagenomics, to assess the possible role of bacteria in OSCC. While these studies have demonstrated differences in microbial composition between cancerous and healthy tissues, they have failed to agree on specific bacteria or patterns of oral microbial dysbiosis to implicate in OSCC. On the contrary, some oral taxa, particularly Porphyromonas gingivalis and Fusobacterium nucleatum, show strong oral carcinogenic potential in vitro and in animal studies. Bacteria are thought to contribute to oral carcinogenesis via inhibition of apoptosis, activation of cell proliferation, promotion of cellular invasion, induction of chronic inflammation, and production of carcinogens. This narrative review provides a critical analysis of and an update on the association between bacteria and oral carcinogenesis and the possible mechanisms underlying it.

  20. Intake of Meat Proteins Substantially Increased the Relative Abundance of Genus Lactobacillus in Rat Feces.

    Science.gov (United States)

    Zhu, Yingying; Lin, Xisha; Li, He; Li, Yingqiu; Shi, Xuebin; Zhao, Fan; Xu, Xinglian; Li, Chunbao; Zhou, Guanghong

    2016-01-01

    Diet has been shown to have a critical influence on gut bacteria and host health, and high levels of red meat in diet have been shown to increase colonic DNA damage and thus be harmful to gut health. However, previous studies focused more on the effects of meat than of meat proteins. In order to investigate whether intake of meat proteins affects the composition and metabolic activities of gut microbiota, feces were collected from growing rats that were fed with either meat proteins (from beef, pork or fish) or non-meat proteins (casein or soy) for 14 days. The resulting composition of gut microbiota was profiled by sequencing the V4-V5 region of the 16S ribosomal RNA genes and the short chain fatty acids (SCFAs) were analyzed using gas chromatography. The composition of gut microbiota and SCFA levels were significantly different between the five diet groups. At a recommended dose of 20% protein in the diet, meat protein-fed rats had a higher relative abundance of the beneficial genus Lactobacillus, but lower levels of SCFAs and SCFA-producing bacteria including Fusobacterium, Bacteroides and Prevotella, compared with the soy protein-fed group. Further work is needed on the regulatory pathways linking dietary protein intake to gut microbiota.

  1. The effect of photodynamic therapy on pathogenic bacteria around peri-implant sulcus and in the cavity between abutment and implant after healing phase: A prospective clinical study.

    Science.gov (United States)

    Zhou, Lin-Yi; Shi, Jun-Yu; Zhu, Yu; Qian, Shu-Jiao; Lai, Hong-Chang; Gu, Ying-Xin

    2018-05-14

    To compare levels of pathogens from peri-implant sulcus versus abutment screw cavities after photodynamic therapy. Twenty patients were included. Photodynamic therapy (PDT) was applied both in sulcus and cavities after sampling following suprastructures loading, and repeated after 2 weeks. Two samples each containing four paper points were collected for each implant at baseline, 2 weeks, 3 months: (i) peri-implant sulcus and (ii) abutment screw cavities. Seventy-five percent ethanol was applied in another 20 patients as the control group in the same way. qPCR was used to quantify periodontal pathogens: Porphyromonas gingivalis, Fusobacterium nucleatum, Streptococcus mutans. PDT showed a better bacterial reduction than ethanol. P. g. and F. n. were most frequently detected, while less for S. m. P. gingivalis' proportion from both sites was significantly higher than the other two bacteria (P abutment screw cavities were always less than those from peri-implant sulcus and was significantly lower for total bacteria at 3 months (P abutment screw cavities significantly reduced at 3 months compared to baseline (P abutment screw cavities in the long run, suggesting PDT an effective way sterilizing inner surface of oral implant suprastrutures. Lasers Surg. Med. © 2018 Wiley Periodicals, Inc. © 2018 Wiley Periodicals, Inc.

  2. Pyrosequencing of Plaque Microflora In Twin Children with Discordant Caries Phenotypes.

    Directory of Open Access Journals (Sweden)

    Meng Zhang

    Full Text Available Despite recent successes in the control of dental caries, the mechanism of caries development remains unclear. To investigate the causes of dental decay, especially in early childhood caries, the supragingival microflora composition of 20 twins with discordant caries phenotypes were analyzed using high-throughput pyrosequencing. In addition, the parents completed a lifestyle questionnaire. A total of 228,789 sequencing reads revealed 10 phyla, 84 genera, and 155 species of microflora, the relative abundances of these strains varied dramatically among the children, Comparative analysis between groups revealed that Veillonella, Corynebacterium and Actinomyces were presumed to be caries-related genera, Fusobacterium, Kingella and Leptotrichia were presumed to be healthy-related genus, yet this six genera were not statistically significant (P>0.05. Moreover, a cluster analysis revealed that the microbial composition of samples in the same group was often dissimilar but that the microbial composition observed in twins was usually similar. Although the genetic and environmental factors that strongly influence the microbial composition of dental caries remains unknown, we speculate that genetic factors primarily influence the individual's susceptibility to dental caries and that environmental factors primarily regulate the microbial composition of the dental plaque and the progression to caries. By using improved twins models and increased sample sizes, our study can be extended to analyze the specific genetic and environmental factors that affect the development of caries.

  3. Effect of Aging on Periodontal Inflammation, Microbial Colonization, and Disease Susceptibility.

    Science.gov (United States)

    Wu, Y; Dong, G; Xiao, W; Xiao, E; Miao, F; Syverson, A; Missaghian, N; Vafa, R; Cabrera-Ortega, A A; Rossa, C; Graves, D T

    2016-04-01

    Periodontitis is a chronic inflammatory disease induced by a biofilm that forms on the tooth surface. Increased periodontal disease is associated with aging. We investigated the effect of aging on challenge by oral pathogens, examining the host response, colonization, and osteoclast numbers in aged versus young mice. We also compared the results with mice with lineage-specific deletion of the transcription factor FOXO1, which reduces dendritic cell (DC) function. Periodontitis was induced by oral inoculation of Porphyromonas gingivalis and Fusobacterium nucleatum in young (4 to 5 mo) and aged (14 to 15 mo) mice. Aged mice as well as mice with reduced DC function had decreased numbers of DCs in lymph nodes, indicative of a diminished host response. In vitro studies suggest that reduced DC numbers in lymph nodes of aged mice may involve the effect of advanced glycation end products on DC migration. Surprisingly, aged mice but not mice with genetically altered DC function had greater production of antibody to P. gingivalis, greater IL-12 expression, and more plasma cells in lymph nodes following oral inoculation as compared with young mice. The greater adaptive immune response in aged versus young mice was linked to enhanced levels of P. gingivalis and reduced bacterial diversity. Thus, reduced bacterial diversity in aged mice may contribute to increased P. gingivalis colonization following inoculation and increased periodontal disease susceptibility, reflected by higher TNF levels and osteoclast numbers in the periodontium of aged versus young mice. © International & American Associations for Dental Research 2016.

  4. Intake of Meat Proteins Substantially Increased the Relative Abundance of Genus Lactobacillus in Rat Feces

    Science.gov (United States)

    Zhu, Yingying; Lin, Xisha; Li, He; Li, Yingqiu; Shi, Xuebin; Zhao, Fan; Xu, Xinglian; Li, Chunbao; Zhou, Guanghong

    2016-01-01

    Diet has been shown to have a critical influence on gut bacteria and host health, and high levels of red meat in diet have been shown to increase colonic DNA damage and thus be harmful to gut health. However, previous studies focused more on the effects of meat than of meat proteins. In order to investigate whether intake of meat proteins affects the composition and metabolic activities of gut microbiota, feces were collected from growing rats that were fed with either meat proteins (from beef, pork or fish) or non-meat proteins (casein or soy) for 14 days. The resulting composition of gut microbiota was profiled by sequencing the V4-V5 region of the 16S ribosomal RNA genes and the short chain fatty acids (SCFAs) were analyzed using gas chromatography. The composition of gut microbiota and SCFA levels were significantly different between the five diet groups. At a recommended dose of 20% protein in the diet, meat protein-fed rats had a higher relative abundance of the beneficial genus Lactobacillus, but lower levels of SCFAs and SCFA-producing bacteria including Fusobacterium, Bacteroides and Prevotella, compared with the soy protein-fed group. Further work is needed on the regulatory pathways linking dietary protein intake to gut microbiota. PMID:27042829

  5. The antimicrobial efficacy of commercial dentifrices.

    Science.gov (United States)

    Haraszthy, Violet I; Zambon, Joseph J; Sreenivasan, Prem K

    2010-01-01

    This investigation compared the effects of a fluoride dentifrice and toothpastes formulated with antimicrobial ingredients (stannous fluoride and triclosan/copolymer) on oral micro-organisms, including those found in samples taken from the human oral cavity. Microbiological techniques determined the minimum inhibitory concentrations (MICs) of each dentifrice necessary to inhibit the growth of bacterial strains from the healthy oral cavity, as well as those found in dental caries, periodontal disease, and halitosis. Ex vivo studies utilized oral rinse samples and supragingival plaque from adults to determine antimicrobial effects on the entire microbial diversity of these samples, including biofilm-derived micro-organisms. The triclosan/copolymer dentifrice demonstrated the lowest MICs and significantly inhibited Gram-positive and Gram-negative bacteria (including the periodontal pathogens Aggregatibacter actinomycetemcomitans, Eikenella corrodens, and Fusobacterium nucleatum). In the ex vivo tests, the triclosan/copolymer dentifrice demonstrated substantial inhibition in the oral rinse samples over each treatment period (p > 0.0005) as compared to either the fluoride or stannous fluoride dentifrices. Similarly, the triclosan/copolymer dentifrice demonstrated the highest inhibition of micro-organisms in the supragingival plaque biofilm (p fluoride and stannous fluoride dentifrices (p > 0.5).

  6. Sequential colonization of periodontal pathogens in induction of periodontal disease and atherosclerosis in LDLRnull mice.

    Science.gov (United States)

    Chukkapalli, Sasanka S; Easwaran, Meena; Rivera-Kweh, Mercedes F; Velsko, Irina M; Ambadapadi, Sriram; Dai, Jiayin; Larjava, Hannu; Lucas, Alexandra R; Kesavalu, Lakshmyya

    2017-01-01

    Periodontal disease (PD) and atherosclerotic vascular disease (ASVD) are both chronic inflammatory diseases with a polymicrobial etiology and have been epidemiologically associated. The purpose is to examine whether periodontal bacteria that infect the periodontium can also infect vascular tissues and enhance pre-existing early aortic atherosclerotic lesions in LDLRnull mice. Mice were orally infected with intermediate bacterial colonizer Fusobacterium nucleatum for the first 12 weeks followed by late bacterial colonizers (Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia) for the remaining 12 weeks mimicking the human oral microbiota ecological colonization. Genomic DNA from all four bacterial was detected in gingival plaque by PCR, consistently demonstrating infection of mouse gingival surfaces. Infected mice had significant levels of IgG and IgM antibodies, alveolar bone resorption, and showed apical migration of junctional epithelium revealing the induction of PD. These results support the ability of oral bacteria to cause PD in mice. Detection of bacterial genomic DNA in systemic organs indicates hematogenous dissemination from the gingival pockets. Bacterial infection did not alter serum lipid fractions or serum amyloid A levels and did not induce aortic atherosclerotic plaque. This is the first study examining the causal role of periodontal bacteria in induction of ASVD in LDLRnull mice. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. DNA methylation differentially regulates cytokine secretion in gingival epithelia in response to bacterial challenges.

    Science.gov (United States)

    Drury, Jeanie L; Chung, Whasun Oh

    2015-03-01

    Epigenetic modifications are changes in gene expression without altering DNA sequence. We previously reported that bacteria-specific innate immune responses are regulated by epigenetic modifications. Our hypothesis is that DNA methylation affects gingival cytokine secretion in response to bacterial stimulation. Gingival epithelial cells (GECs) were treated with DNMT-1 inhibitors prior to Porphyromonas gingivalis (Pg) or Fusobacterium nucleatum (Fn) exposure. Protein secretion was assessed using ELISA. Gene expression was quantified using qRT-PCR. The ability of bacteria to invade inhibitor pretreated GECs was assessed utilizing flow cytometry. Changes were compared to unstimulated GECs. GEC upregulation of IL-6 and CXCL1 by Pg or Fn stimulation was significantly diminished by inhibitor pretreatment. Pg stimulated IL-1α secretion and inhibitor pretreatment significantly enhanced this upregulation, while Fn alone or with inhibitor pretreatment had no effect on IL-1α expression. GEC upregulation of human beta-definsin-2 in response to Pg and Fn exposure was enhanced following the inhibitor pretreatment. GEC susceptibility to bacterial invasion was unaltered. These results suggest that DNA methylation differentially affects gingival cytokine secretion in response to Pg or Fn. Our data provide basis for better understanding of how epigenetic modifications, brought on by exposure to oral bacteria, will subsequently affect host susceptibility to oral diseases. © FEMS 2014. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Comparative evaluation of antiplaque and antigingivitis effects of an herbal and chlorine dioxide mouthwashes: A clinicomicrobiological study

    Directory of Open Access Journals (Sweden)

    Srinivasa T Siddeshappa

    2018-01-01

    Full Text Available Aim: The aim of the present study was to compare the efficacy of herbal mouthwash and chlorine dioxide mouthwash in reduction of plaque and gingivitis. Settings and Design: In a randomized clinical trial, forty patients were randomly selected and divided equally into two groups. Materials and Methods: After professional oral prophylaxis, the clinical parameters plaque index, gingival index, and modified sulcular bleeding index were recorded at baseline, 7th day, 14th day, and 21st day. The plaque samples were collected from gingival sulcus with an absorbent sterile paper point and were stored in a thioglycollate broth, then sent for microbiological examination. The microbial colony-forming units were assessed at baseline, 7th day, 14th day, and 21st day for Streptococcus mutans, Tannerella forsythia, and Fusobacterium nucleatum. Results: There was a statistical significant reduction in both clinical and microbiological parameters were observed with use of both the mouthwashes. However, herbal mouthwash was more effective in reducing the plaque and gingivitis than chlorine dioxide mouthwash. Conclusion: Herbal mouthwash was statistically efficacious in controlling plaque and gingivitis with potent antimicrobial activity.

  9. Microbiological, lipid and immunological profiles in children with gingivitis and type 1 diabetes mellitus.

    Science.gov (United States)

    Duque, Cristiane; João, Mariana Ferreira Dib; Camargo, Gabriela Alessandra da Cruz Galhardo; Teixeira, Gláucia Schuindt; Machado, Thamiris Santana; Azevedo, Rebeca de Souza; Mariano, Flávia Sammartino; Colombo, Natália Helena; Vizoto, Natália Leal; Mattos-Graner, Renata de Oliveira

    2017-01-01

    The aim of this study was to compare the prevalence of periodontal pathogens, systemic inflammatory mediators and lipid profiles in type 1 diabetes children (DM) with those observed in children without diabetes (NDM), both with gingivitis. Twenty-four DM children and twenty-seven NDM controls were evaluated. The periodontal status, glycemic and lipid profiles were determined for both groups. Subgingival samples of periodontal sites were collected to determine the prevalence of periodontal microorganisms by PCR. Blood samples were collected for IL-1-β, TNF-α and IL-6 analysis using ELISA kits. Periodontal conditions of DM and NDM patients were similar, without statistical differences in periodontal indices. When considering patients with gingivitis, all lipid parameters evaluated were highest in the DM group; Capnocytophaga sputigena and Capnocytophaga ochracea were more prevalent in the periodontal sites of DM children. "Red complex" bacteria were detected in few sites of DM and NDM groups. Fusobacterium nucleatum and Campylobacter rectus were frequently found in both groups. Similar levels of IL-1-β, TNF-α and IL-6 were detected in DM and NDM children. Clinical and immunological profiles are similar between DM and NDM children. The presence of Capnocytophaga sputigena and Capnocytophaga ochracea were associated with gingivitis in DM children.

  10. Microbial Composition Predicts Genital Tract Inflammation and Persistent Bacterial Vaginosis in South African Adolescent Females.

    Science.gov (United States)

    Lennard, Katie; Dabee, Smritee; Barnabas, Shaun L; Havyarimana, Enock; Blakney, Anna; Jaumdally, Shameem Z; Botha, Gerrit; Mkhize, Nonhlanhla N; Bekker, Linda-Gail; Lewis, David A; Gray, Glenda; Mulder, Nicola; Passmore, Jo-Ann S; Jaspan, Heather B

    2018-01-01

    Young African females are at an increased risk of HIV acquisition, and genital inflammation or the vaginal microbiome may contribute to this risk. We studied these factors in 168 HIV-negative South African adolescent females aged 16 to 22 years. Unsupervised clustering of 16S rRNA gene sequences revealed three clusters (subtypes), one of which was strongly associated with genital inflammation. In a multivariate model, the microbiome compositional subtype and hormonal contraception were significantly associated with genital inflammation. We identified 40 taxa significantly associated with inflammation, including those reported previously ( Prevotella , Sneathia , Aerococcus , Fusobacterium , and Gemella ) as well as several novel taxa (including increased frequencies of bacterial vaginosis-associated bacterium 1 [BVAB1], BVAB2, BVAB3, Prevotella amnii , Prevotella pallens , Parvimonas micra , Megasphaera , Gardnerella vaginalis , and Atopobium vaginae and decreased frequencies of Lactobacillus reuteri , Lactobacillus crispatus , Lactobacillus jensenii , and Lactobacillus iners ). Women with inflammation-associated microbiomes had significantly higher body mass indices and lower levels of endogenous estradiol and luteinizing hormone. Community functional profiling revealed three distinct vaginal microbiome subtypes, one of which was characterized by extreme genital inflammation and persistent bacterial vaginosis (BV); this subtype could be predicted with high specificity and sensitivity based on the Nugent score (≥9) or BVAB1 abundance. We propose that women with this BVAB1-dominated subtype may have chronic genital inflammation due to persistent BV, which may place them at a particularly high risk for HIV infection. Copyright © 2017 American Society for Microbiology.

  11. Subgingival Microbiota in White Patients With Desquamative Gingivitis: A Cross-Sectional Study.

    Science.gov (United States)

    Arduino, Paolo G; Romano, Federica; Sasia, Danilo; Broccoletti, Roberto; Ricceri, Fulvio; Barbui, Anna Maria; Brossa, Silvia; Cipriani, Raffaella; Cricenti, Luca; Cabras, Marco; Aimetti, Mario

    2017-07-01

    Presence of epithelial desquamation, erythema, and erosions on gingival tissue is usually described in the literature as desquamative gingivitis (DG). A wide range of autoimmune/dermatologic disorders can manifest as DG, although the two more common are oral lichen planus and mucous membrane pemphigoid. The aim of this study is to investigate prevalence of 11 periodontopathogenic microorganisms in patients with DG and to compare it with the microbiologic status of individuals affected by plaque-induced gingivitis (pGI). Cross-sectional clinical and microbiologic data were obtained from 66 patients (33 in each group). Subgingival plaque samples were analyzed using semiquantitative polymerase chain reaction analysis. Statistically significant difference, but with little clinical significance, was observed in gingival conditions between the two groups, probably due to the worse home control hygiene of patients with DG. Prevalence and levels of Aggregatibacter actinomycetemcomitans, Eikenella corrodens, and Fusobacterium nucleatum/periodonticum were statistically higher in samples from patients with DG than in those with pGI. In multivariate regression models, subgingival colonization of A. actinomycetemcomitans and F. nucleatum/periodonticum was not statistically associated with DG, whereas, high levels of E. corrodens were associated with 13-fold increased odds for DG. Microbiologic differences were found in subgingival plaque for patients with DG and pGI. This may suggest possible association between periodontal pathogens and DG.

  12. Comparative profiling of microbial community of three economically important fishes reared in sea cages under tropical offshore environment.

    Science.gov (United States)

    Rasheeda, M K; Rangamaran, Vijaya Raghavan; Srinivasan, Senthilkumar; Ramaiah, Sendhil Kumar; Gunasekaran, Rajaprabhu; Jaypal, Santhanakumar; Gopal, Dharani; Ramalingam, Kirubagaran

    2017-08-01

    The present study was undertaken to evaluate the microbial composition of farmed cobia pompano and milkfish, reared in sea-cages by culture-independent methods. This study would serve as a basis for assessing the general health of fish, identifying the dominant bacterial species present in the gut for future probiotic work and in early detection of potential pathogens. High-throughput sequencing of V3-V4 hyper variable regions of 16S rDNA on Illumina MiSeq platform facilitated unravelling of composite bacterial population. Analysis of 1.3 million quality-filtered sequences revealed high microbial diversity. Characteristic marine fish gut microbes: Vibrio and Photobacterium spp. showed prevalence in cobia and pompano whereas Pelomonas and Fusobacterium spp. dominated the gut of milkfish. Pompano hindgut with 10,537 operational taxonomy units (OTUs) exhibited the highest alpha-diversity index followed by cobia (10,435) and milkfish (2799). Additionally unique and shared OTUs in each gut type were identified. Gammaproteobacteria dominated in cobia and pompano while Betaproteobacteria showed prevalence in milkfish. We obtained 96 shared OTUs among the three species though the numbers of reads were highly variable. These differences in microbiota of farmed fish reared in same environment were presumably due to differences in the gut morphology, physiological behavior and host specificity. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Streptococcus pneumoniae Colonization Is Required To Alter the Nasal Microbiota in Cigarette Smoke-Exposed Mice.

    Science.gov (United States)

    Shen, Pamela; Whelan, Fiona J; Schenck, L Patrick; McGrath, Joshua J C; Vanderstocken, Gilles; Bowdish, Dawn M E; Surette, Michael G; Stämpfli, Martin R

    2017-10-01

    Smokers have nasal microbiota dysbiosis, with an increased frequency of colonizing bacterial pathogens. It is possible that cigarette smoke increases pathogen acquisition by perturbing the microbiota and decreasing colonization resistance. However, it is difficult to disentangle microbiota dysbiosis due to cigarette smoke exposure from microbiota changes caused by increased pathogen acquisition in human smokers. Using an experimental mouse model, we investigated the impact of cigarette smoke on the nasal microbiota in the absence and presence of nasal pneumococcal colonization. We observed that cigarette smoke exposure alone did not alter the nasal microbiota composition. The microbiota composition was also unchanged at 12 h following low-dose nasal pneumococcal inoculation, suggesting that the ability of the microbiota to resist initial nasal pneumococcal acquisition was not impaired in smoke-exposed mice. However, nasal microbiota dysbiosis occurred as a consequence of established high-dose nasal pneumococcal colonization at day 3 in smoke-exposed mice. Similar to clinical reports on human smokers, an enrichment of potentially pathogenic bacterial genera such as Fusobacterium , Gemella , and Neisseria was observed. Our findings suggest that cigarette smoke exposure predisposes to pneumococcal colonization independent of changes to the nasal microbiota and that microbiota dysbiosis observed in smokers may occur as a consequence of established pathogen colonization. Copyright © 2017 American Society for Microbiology.

  14. In Silico Analysis of Putrefaction Pathways in Bacteria and Its Implication in Colorectal Cancer

    Directory of Open Access Journals (Sweden)

    Harrisham Kaur

    2017-11-01

    Full Text Available Fermentation of undigested proteins in human gastrointestinal tract (gut by the resident microbiota, a process called bacterial putrefaction, can sometimes disrupt the gut homeostasis. In this process, essential amino acids (e.g., histidine, tryptophan, etc. that are required by the host may be utilized by the gut microbes. In addition, some of the products of putrefaction, like ammonia, putrescine, cresol, indole, phenol, etc., have been implicated in the disease pathogenesis of colorectal cancer (CRC. We have investigated bacterial putrefaction pathways that are known to be associated with such metabolites. Results of the comprehensive in silico analysis of the selected putrefaction pathways across bacterial genomes revealed presence of these pathways in limited bacterial groups. Majority of these bacteria are commonly found in human gut. These include Bacillus, Clostridium, Enterobacter, Escherichia, Fusobacterium, Salmonella, etc. Interestingly, while pathogens utilize almost all the analyzed pathways, commensals prefer putrescine and H2S production pathways for metabolizing the undigested proteins. Further, comparison of the putrefaction pathways in the gut microbiomes of healthy, carcinoma and adenoma datasets indicate higher abundances of putrefying bacteria in the carcinoma stage of CRC. The insights obtained from the present study indicate utilization of possible microbiome-based therapies to minimize the adverse effects of gut microbiome in enteric diseases.

  15. In Vitro Evaluation of Planktonic Growth on Experimental Cement-Retained Titanium Surfaces.

    Science.gov (United States)

    Balci, Nur; Cakan, Umut; Aksu, Burak; Akgul, Oncu; Ulger, Nurver

    2016-04-08

    BACKGROUND The purpose of this study was to compare the effects of selected cements, or their combination with titanium, on the growth of two periodontopathic bacteria: Prevotella intermedia (Pi) and Fusobacterium nucleatum (Fn). MATERIAL AND METHODS This study was comprised of several experimental groups: 1) Dental luting cements (glass ionomer cement, methacrylate-based resin cement, zinc-oxide eugenol cement, eugenol-free zinc oxide cement; 2) titanium discs; and 3) titanium combination cement discs. The disks were submerged in bacterial suspensions of either Fn or Pi. Planktonic bacterial growth within the test media was measured by determining the optical density of the cultures (OD600). Mean and standard deviations were calculated for planktonic growth from three separate experiments. RESULTS Intergroup comparison of all experimental groups revealed increased growth of Pi associated with cement-titanium specimens in comparison with cement specimens. Regarding the comparison of all groups for Fn, there was an increased amount of bacterial growth in cement-titanium specimens although the increase was not statistically significant. CONCLUSIONS The combination of cement with titanium may exacerbate the bacterial growth capacity of Pi and Fn in contrast to their sole effect.

  16. Role of Cathepsin S in Periodontal Inflammation and Infection

    Directory of Open Access Journals (Sweden)

    S. Memmert

    2017-01-01

    Full Text Available Cathepsin S is a cysteine protease and regulator of autophagy with possible involvement in periodontitis. The objective of this study was to investigate whether cathepsin S is involved in the pathogenesis of periodontal diseases. Human periodontal fibroblasts were cultured under inflammatory and infectious conditions elicited by interleukin-1β and Fusobacterium nucleatum, respectively. An array-based approach was used to analyze differential expression of autophagy-associated genes. Cathepsin S was upregulated most strongly and thus further studied in vitro at gene and protein levels. In vivo, gingival tissue biopsies from rats with ligature-induced periodontitis and from periodontitis patients were also analyzed at transcriptional and protein levels. Multiple gene expression changes due to interleukin-1β and F. nucleatum were observed in vitro. Both stimulants caused a significant cathepsin S upregulation. A significantly elevated cathepsin S expression in gingival biopsies from rats with experimental periodontitis was found in vivo, as compared to that from control. Gingival biopsies from periodontitis patients showed a significantly higher cathepsin S expression than those from healthy gingiva. Our findings provide original evidence that cathepsin S is increased in periodontal cells and tissues under inflammatory and infectious conditions, suggesting a critical role of this autophagy-associated molecule in the pathogenesis of periodontitis.

  17. Comparative Study on the Characteristics of Weissella cibaria CMU and Probiotic Strains for Oral Care

    Directory of Open Access Journals (Sweden)

    Hye-Jin Jang

    2016-12-01

    Full Text Available Probiotics have been demonstrated as a new paradigm to substitute antibiotic treatment for dental caries, gingivitis, and chronic periodontitis. The present work was conducted to compare the characteristics of oral care probiotics: Weissella cibaria CMU (Chonnam Medical University and four commercial probiotic strains. Survival rates under poor oral conditions, acid production, hydrogen peroxide production, as well as inhibition of biofilm formation, coaggregation, antibacterial activity, and inhibition of volatile sulfur compounds were evaluated. The viability of W. cibaria CMU was not affected by treatment of 100 mg/L lysozyme for 90 min and 1 mM hydrogen peroxide for 6 h. Interestingly, W. cibaria produced less acid and more hydrogen peroxide than the other four probiotics. W. cibaria inhibited biofilm formation by Streptococcus mutans at lower concentrations (S. mutans/CMU = 8 and efficiently coaggregated with Fusobacterium nucleatum. W. cibaria CMU and two commercial probiotics, including Lactobacillus salivarius and Lactobacillus reuteri, showed high antibacterial activities (>97% against cariogens (S. mutans and Streptococcus sobrinus, and against periodontopathogens (F. nucleatum and Porphyromonas gingivalis. All of the lactic acid bacterial strains in this study significantly reduced levels of hydrogen sulfide and methyl mercaptan produced by F. nucleatum and P. gingivalis (p < 0.05. These results suggest that W. cibaria CMU is applicable as an oral care probiotic.

  18. The Use of Different Irrigation Techniques to Decrease Bacterial Loads in Healthy and Diabetic Patients with Asymptomatic Apical Periodontitis.

    Science.gov (United States)

    Ghoneim, Mai; Saber, Shehab ElDin; El-Badry, Tarek; Obeid, Maram; Hassib, Nehal

    2016-12-15

    Diabetes mellitus is a multisystem disease which weakens the human's immunity. Subsequently, it worsens the sequelae of apical periodontitis by raising a fierce bacterial trait due to the impaired host response. This study aimed to estimate bacterial reduction after using different irrigation techniques in systemically healthy and diabetic patients with asymptomatic apical periodontitis. Enterococcus faecalis , Peptostreptococcus micros , and Fusobacterium necleatum bacteria were chosen, as they are the most common and prevailing strains found in periodontitis. Bacterial samples were retrieved from necrotic root canals of systemically healthy and diabetic patients, before and after endodontic cleaning and shaping by using two different irrigation techniques; the conventional one and the EndoVac system. Quantitive polymerase chain reaction (qPCR) was utilised to detect the reduction in the bacterial count. The EndoVac irrigation system was effective in reducing bacteria, especially Peptostreptococcus micros in the diabetic group when compared to conventional irrigation technique with a statistically significant difference. The EndoVac can be considered as a promising tool in combination with irrigant solution to defeat the bacterial colonies living in the root canal system. Additional studies ought to be done to improve the means of bacterial clearance mainly in immune-compromised individuals.

  19. Environmental Arsenic Exposure and Microbiota in Induced Sputum

    Directory of Open Access Journals (Sweden)

    Allison G. White

    2014-02-01

    Full Text Available Arsenic exposure from drinking water is associated with adverse respiratory outcomes, but it is unknown whether arsenic affects pulmonary microbiota. This exploratory study assessed the effect of exposure to arsenic in drinking water on bacterial diversity in the respiratory tract of non-smokers. Induced sputum was collected from 10 subjects with moderate mean household water arsenic concentration (21.1 ± 6.4 ppb and 10 subjects with low household water arsenic (2.4 ± 0.8 ppb. To assess microbiota in sputum, the V6 hypervariable region amplicons of bacterial 16s rRNA genes were sequenced using the Ion Torrent Personal Genome Machine. Microbial community differences between arsenic exposure groups were evaluated using QIIME and Metastats. A total of 3,920,441 sequence reads, ranging from 37,935 to 508,787 per sample for 316 chips after QIIME quality filtering, were taxonomically classified into 142 individual genera and five phyla. Firmicutes (22%, Proteobacteria (17% and Bacteriodetes (12% were the main phyla in all samples, with Neisseriaceae (15%, Prevotellaceae (12% and Veillonellacea (7% being most common at the genus level. Some genera, including Gemella, Lactobacillales, Streptococcus, Neisseria and Pasteurellaceae were elevated in the moderate arsenic exposure group, while Rothia, Prevotella, Prevotellaceae Fusobacterium and Neisseriaceae were decreased, although none of these differences was statistically significant. Future studies with more participants and a greater range of arsenic exposure are needed to further elucidate the effects of drinking water arsenic consumption on respiratory microbiota.

  20. Putative periodontopathic bacteria and herpesviruses in pregnant women: a case-control study.

    Science.gov (United States)

    Lu, Haixia; Zhu, Ce; Li, Fei; Xu, Wei; Tao, Danying; Feng, Xiping

    2016-06-15

    Little is known about herpesvirus and putative periodontopathic bacteria in maternal chronic periodontitis. The present case-control study aimed to explore the potential relationship between putative periodontopathic bacteria and herpesviruses in maternal chronic periodontitis.Saliva samples were collected from 36 pregnant women with chronic periodontitis (cases) and 36 pregnant women with healthy periodontal status (controls). Six putative periodontopathic bacteria (Porphyromonas gingivalis [Pg], Aggregatibacer actinomycetemcomitans [Aa], Fusobacterium nucleatum [Fn], Prevotella intermedia [Pi], Tannerella forsythia [Tf], and Treponema denticola [Td]) and three herpesviruses (Epstein-Barr virus [EBV], human cytomegalovirus [HCMV], and herpes simplex virus [HSV]) were detected. Socio-demographic data and oral health related behaviors, and salivary estradiol and progesterone levels were also collected. The results showed no significant differences in socio-demographic background, oral health related behaviors, and salivary estradiol and progesterone levels between the two groups (all P > 0.05). The detection rates of included periodontopathic microorganisms were not significantly different between the two groups (all P > 0.05), but the coinfection rate of EBV and Pg was significantly higher in the case group than in the control group (P = 0.028). EBV and Pg coinfection may promote the development of chronic periodontitis among pregnant women.

  1. Prevotella intermedia induces severe bacteremic pneumococcal pneumonia in mice with upregulated platelet-activating factor receptor expression.

    Science.gov (United States)

    Nagaoka, Kentaro; Yanagihara, Katsunori; Morinaga, Yoshitomo; Nakamura, Shigeki; Harada, Tatsuhiko; Hasegawa, Hiroo; Izumikawa, Koichi; Ishimatsu, Yuji; Kakeya, Hiroshi; Nishimura, Masaharu; Kohno, Shigeru

    2014-02-01

    Streptococcus pneumoniae is the leading cause of respiratory infection worldwide. Although oral hygiene has been considered a risk factor for developing pneumonia, the relationship between oral bacteria and pneumococcal infection is unknown. In this study, we examined the synergic effects of Prevotella intermedia, a major periodontopathic bacterium, on pneumococcal pneumonia. The synergic effects of the supernatant of P. intermedia (PiSup) on pneumococcal pneumonia were investigated in mice, and the stimulation of pneumococcal adhesion to human alveolar (A549) cells by PiSup was assessed. The effects of PiSup on platelet-activating factor receptor (PAFR) transcript levels in vitro and in vivo were analyzed by quantitative real-time PCR, and the differences between the effects of pneumococcal infection induced by various periodontopathic bacterial species were verified in mice. Mice inoculated with S. pneumoniae plus PiSup exhibited a significantly lower survival rate, higher bacterial loads in the lungs, spleen, and blood, and higher inflammatory cytokine levels in the bronchoalveolar lavage fluid (macrophage inflammatory protein 2 and tumor necrosis factor alpha) than those infected without PiSup. In A549 cells, PiSup increased pneumococcal adhesion and PAFR transcript levels. PiSup also increased lung PAFR transcript levels in mice. Similar effects were not observed in the supernatants of Porphyromonas gingivalis or Fusobacterium nucleatum. Thus, P. intermedia has the potential to induce severe bacteremic pneumococcal pneumonia with enhanced pneumococcal adhesion to lower airway cells.

  2. Nucleases from Prevotella intermedia can degrade neutrophil extracellular traps.

    Science.gov (United States)

    Doke, M; Fukamachi, H; Morisaki, H; Arimoto, T; Kataoka, H; Kuwata, H

    2017-08-01

    Periodontitis is an inflammatory disease caused by periodontal bacteria in subgingival plaque. These bacteria are able to colonize the periodontal region by evading the host immune response. Neutrophils, the host's first line of defense against infection, use various strategies to kill invading pathogens, including neutrophil extracellular traps (NETs). These are extracellular net-like fibers comprising DNA and antimicrobial components such as histones, LL-37, defensins, myeloperoxidase, and neutrophil elastase from neutrophils that disarm and kill bacteria extracellularly. Bacterial nuclease degrades the NETs to escape NET killing. It has now been shown that extracellular nucleases enable bacteria to evade this host antimicrobial mechanism, leading to increased pathogenicity. Here, we compared the DNA degradation activity of major Gram-negative periodontopathogenic bacteria, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans. We found that Pr. intermedia showed the highest DNA degradation activity. A genome search of Pr. intermedia revealed the presence of two genes, nucA and nucD, putatively encoding secreted nucleases, although their enzymatic and biological activities are unknown. We cloned nucA- and nucD-encoding nucleases from Pr. intermedia ATCC 25611 and characterized their gene products. Recombinant NucA and NucD digested DNA and RNA, which required both Mg 2+ and Ca 2+ for optimal activity. In addition, NucA and NucD were able to degrade the DNA matrix comprising NETs. © 2016 The Authors Molecular Oral Microbiology Published by John Wiley & Sons Ltd.

  3. Beneficial Oral Biofilms as Smart Bioactive Interfaces

    Directory of Open Access Journals (Sweden)

    Beatrice Gutt

    2018-01-01

    Full Text Available Periodontitis is a very common health problem caused by formation of pathogenic bacterial biofilm that triggers inflammation resulting in either reversible gingivitis or irreversible periodontal hard and soft tissue damages, leading to loss of teeth when left untreated. Commensal bacteria play an important role in oral health in many aspects. Mainly by colonizing oral tissues, they (i contribute to maturation of immune response, and (ii foreclose attachment of pathobiont and, therefore, prevent from infection. The main goal of the study was to investigate if blocking of receptors on a commensal biofilm can prevent or reduce the attachment of pathogenic strains. To do so, biofilm produced by commensal Streptococcus sanguinis was treated with whole cell lysate of pathobionts Fusobacterium nucleatum or Porphyromonas gingivalis, followed by incubation with respective strain(s. The study revealed significant reduction in pathobiont adhesion to lysate-treated commensal biofilm. Therefore, adhesion of pathobionts onto the lysate-blocked biofilm was hindered; however, not completely eliminated supporting the idea that such approach in the oral cavity would benefit the production of a well-balanced and healthy bioactive interface.

  4. Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

    International Nuclear Information System (INIS)

    Gangi Setty, Thanuja; Cho, Christine; Govindappa, Sowmya; Apicella, Michael A.; Ramaswamy, S.

    2014-01-01

    Structure–function studies of sialic acid-binding proteins from F. nucleatum, P. multocida, V. cholerae and H. influenzae reveal a conserved network of hydrogen bonds involved in conformational change on ligand binding. Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states

  5. Molecular-level evaluation of selected periodontal pathogens from subgingival regions in canines and humans with periodontal disease.

    Science.gov (United States)

    Gołyńska, Magdalena; Polkowska, Izabela; Bartoszcze-Tomaszewska, Małgorzata; Sobczyńska-Rak, Aleksandra; Matuszewski, Łukasz

    2017-03-30

    Dogs commonly serve as a model for various human conditions, including periodontal diseases. The aim of this study was to identify the anaerobic bacteria that colonize the subgingival areas in dogs and humans by using rapid real-time polymerase chain reaction (RT-PCR)-based tests and to compare the results obtained in each species. Bacterial microflora evaluations, both quantitative and qualitative, were performed by applying ready-made tests on twelve dogs and twelve humans. Five samples were collected from each subject's deepest gingival pockets and joined to form a collective sample. The results of the study revealed interspecies similarities in the prevalences of Porphyromonas ( P .) gingivalis, Treponema denticola, Tannerella forsythia , and Fusobacterium nucleatum . Red complex bacteria comprised the largest portion of the studied bacterial complexes in all study groups, with P. gingivalis being the most commonly isolated bacterium. The results show similarities in the prevalence of bacterial microflora in dogs and humans. Microbiological analysis of gingival pockets by using rapid real-time PCR-based tests in clinical practice, both veterinary and human, can facilitate the choice of appropriate pharmacological treatment and can provide a basis for subsequent verification of the treatment's effectiveness.

  6. Canine periodontal disease control using a clindamycin hydrochloride gel.

    Science.gov (United States)

    Johnston, Thomas P; Mondal, Pravakar; Pal, Dhananjay; MacGee, Scott; Stromberg, Arnold J; Alur, Hemant

    2011-01-01

    Stabilizing or reducing periodontal pocket depth can have a positive influence on the retention of teeth in dogs. A topical 2% clindamycin hydrochloride gel (CHgel) was evaluated for the treatment of periodontal disease in dogs. The CHgel formulation provides for the sustained erosion of the matrix, but also flows into the periodontal pocket as a viscous liquid, and then rapidly forms a gel that has mucoadhesive properties and also may function as a physical barrier to the introduction of bacteria. A professional teeth cleaning procedure including scaling and root planing was done in dogs with one group receiving CHgel following treatment. Periodontal health was determined before and after the procedure including measurement of periodontal pocket depth, gingival index, gingival bleeding sites, and number of suppurating sites. There was a statistically significant decrease in periodontal pocket depth (19%), gingival index (16%), and the number of bleeding sites (64%) at 90-days in dogs receiving CHgel. Additionally, the number of suppurating sites was lower (93%) at 90-days for the group receiving CHgel. The addition of CHgel effectively controlled the bacterial burden (e.g, Fusobacterium nucleatum) at both day 14 and 90. Gingival cells in culture were shown to rapidly incorporate clindamycin and attain saturation in approximately 20-minutes. In summary, a professional teeth cleaning procedure including root planning and the addition of CHgel improves the gingival index and reduces periodontal pocket depth.

  7. Amixicile, a novel strategy for targeting oral anaerobic pathogens.

    Science.gov (United States)

    Hutcherson, Justin A; Sinclair, Kathryn M; Belvin, Benjamin R; Gui, Qin; Hoffman, Paul S; Lewis, Janina P

    2017-09-05

    The oral microflora is composed of both health-promoting as well as disease-initiating bacteria. Many of the disease-initiating bacteria are anaerobic and include organisms such as Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Tannerella forsythia. Here we investigated a novel therapeutic, amixicile, that targets pyruvate:ferredoxin oxidoreductase (PFOR), a major metabolic enzyme involved in energy generation through oxidative decarboxylation of pyruvate. PFOR is present in these anaerobic pathogenic bacteria and thus we hypothesized that amixicile would effectively inhibit their growth. In general, PFOR is present in all obligate anaerobic bacteria, while oral commensal aerobes, including aerotolerant ones, such as Streptococcus gordonii, use pyruvate dehydrogenase to decarboxylate pyruvate. Accordingly, we observed that growth of the PFOR-containing anaerobic periodontal pathogens, grown in both monospecies as well as multispecies broth cultures was inhibited in a dose-dependent manner while that of S. gordonii was unaffected. Furthermore, we also show that amixicile is effective against these pathogens grown as monospecies and multispecies biofilms. Finally, amixicile is the first selective therapeutic agent active against bacteria internalized by host cells. Together, the results show that amixicile is an effective inhibitor of oral anaerobic bacteria and as such, is a good candidate for treatment of periodontal diseases.

  8. Susceptibility of anaerobic bacteria in Auckland: 1991-1996.

    Science.gov (United States)

    Shore, K P; Pottumarthy, S; Morris, A J

    1999-11-12

    To determine the antimicrobial susceptibility of local anaerobic bacteria. The antimicrobial susceptibility of 357 obligate anaerobes collected between 1991 and 1997 was determined by a standard agar dilution method. Isolates tested included Bacteroides spp. 131, Fusobacterium spp. 12, Prevotella spp. 13, Veillonella spp. 5, Clostridium perfringens 27, other Clostridium spp. 29, Propionibacterium spp. 57, Actinomyces spp. 7, other non-sporing gram-positive bacilli 28 and Peptostreptococcus spp. 48. Ten antimicrobials were tested: penicillin, amoxycillin/ clavulanic acid, pipercillin/tazobactam, ceftriaxone, cefoxitin, cefotetan, imipenem, meropenem, clindamycin and metronidazole. Imipenem, pipercillin/tazobactam, meropenem and amoxycillin/clavulanic acid were active against virtually all anaerobes tested. Metronidazole was active against all anaerobic gram-negative bacteria and Clostridium spp., but had variable activity against other anaerobes. Cefoxitin was the most active cephalosporin against Bacteroides spp., with 76%, 64% and 15% of Bacteroides spp. being susceptible to cefoxitin, cefotetan and ceftriaxone, respectively. Penicillin had poor activity against anaerobic gram negative bacilli. Actinomyces and Propionibacterium spp. were susceptible to all antimicrobials tested except metronidazole. Variable results were obtained with other antimicrobial-organism combinations. Comparison of results with data from a previously published survey showed little change in susceptibility except for increased resistance of Bacteroides fragilis to ceftriaxone and Clostridium species (not C perfringens) to clindamycin. Our results update the local susceptibility profile of anaerobic bacteria and may be considered when choosing an antimicrobial agent for prophylaxis or treatment of anaerobic infections.

  9. Isolation of obligate anaerobic bacteria from ulcerative keratitis in domestic animals.

    Science.gov (United States)

    Ledbetter, Eric C; Scarlett, Janet M

    2008-01-01

    To determine the frequency of obligate anaerobic bacterial isolation from corneal samples of domestic animals with ulcerative keratitis and to characterize the historical, clinical, cytological, and microbiological features of culture-positive cases. Three hundred and thirty domestic animals with ulcerative keratitis. Anaerobic bacteriologic culture and Gram stain were performed on corneal samples from consecutive animals examined with suspect septic ulcerative keratitis. Additional corneal diagnostics included: aerobic bacteriologic culture for all species; fungal culture for ungulates; Mycoplasma culture and virus isolation or feline herpesvirus-1 (FHV-1) polymerase chain reaction (PCR) for cats. Historical, clinical, and cytological findings were correlated with microbiologic data. Anaerobic bacteria were isolated from 13.0% of corneal samples (dogs: 14.0%; horses: 12.9%; cats: 7.9%; alpacas: 18.8%). The most frequent isolates were Clostridium, Peptostreptococcus, Actinomyces, Fusobacterium, and Bacteroides species. The majority of these infections were mixed anaerobic and aerobic bacteria, unless antimicrobial therapy had been administered prior to presentation. The clinical appearance of anaerobic bacterial culture-positive cases was highly variable. Ocular trauma, pre-existing corneal disease, previous corneal surgery, and chronic dermatological disease were significantly (P anaerobic cultures in one or more species. The results of the present study demonstrate that obligate anaerobic bacteria are present within the intralesional flora of ulcerative keratitis in domestic animals. In most species evaluated, these bacteria were identified infrequently. Anaerobic bacterial infection of the cornea most frequently occurs in association with other ocular pathogens and previous corneal abnormalities.

  10. [Role of obligate anaerobes in infections in hospitalized patients and therapeutic options].

    Science.gov (United States)

    Wróblewska, Marta; Rokosz, Alicja; Sawicka-Grzelak, Anna; Kot, Katarzyna; Luczak, Mirosław

    2005-01-01

    Monitoring of prevalence and antibiotic susceptibility of strictly anaerobic bacteria, causing infections in hospitalized patients, constitutes a part of a program for prudent use of antibiotics. The aim of the study was to assess prevalence of strictly anaerobic bacteria in patients hospitalized in a tertiary care hospital in 2001-2002 with reference to empiric antibiotic therapy. The most common gram-positive bacteria were Clostridium difficile--27.7%, Peptostreptococcus spp. and Peptoniphilus asaccharolyticus--21.9% and Actinomyces spp.--11.1%. There was an increase in the number of stool samples positive for C. difficile toxins A and B from 39.4% in 2001 to 59.0% in 2002. The results of susceptibility testing of gram-positive isolates showed the highest percentages of strains susceptible to piperacilin/tazobactam--99.6%, ticarcillin/clavulanate--98.5%, imipenem--98.5%, amoxicillin/clavulanate--97.4% and piperacillin--97.4%. The most prevalent gram-negative anaerobes were strains of Bacteroides spp.--43.1%, Prevotella spp.--35.8% and Fusobacterium spp.--11.0%. All tested strains of gram-negative bacteria were susceptible to metronidazole, piperacilin/tazobactam, ticarcillin/clavulanate and imipenem. In the analyzed population beta-lactam antibiotics with beta-lactamase inhibitors, carbapenems and metronidazole may be used in empiric therapy of infections caused by strictly anaerobic bacteria.

  11. Effects of stress hormones on the production of volatile sulfur compounds by periodontopathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Caroline Morini Calil

    2014-06-01

    Full Text Available Little is known about the effects of stress hormones on the etiologic agents of halitosis. Thus, the aim of this study was to evaluate in vitro the effects of adrenaline (ADR, noradrenaline (NA and cortisol (CORT on bacteria that produce volatile sulfur compounds (VSC, the major gases responsible for bad breath. Cultures of Fusobacterium nucleatum (Fn, Porphyromonas endodontalis (Pe, Prevotella intermedia (Pi and Porphyromonas gingivalis (Pg were exposed to 50 µM ADR, NA and CORT or equivalent volumes of sterile water as controls for 12 and 24 h. Growth was evaluated based on absorbance at 660 nm. Portable gas chromatography was used to measure VSC concentrations. Kruskal-Wallis and the Dunn post-hoc test were used to compare the groups. For Fn, ADR, NA and CORT significantly reduced bacterial growth after 12 h and 24 h (p 0.05. In the Pi cultures, ADR, NA and CORT increased H2S (p < 0.05. Catecholamines and cortisol can interfere with growth and H2S production of sub-gingival species in vitro. This process appears to be complex and supports the association between stress and the production of VSC.

  12. Effects of Hangeshashinto on Growth of Oral Microorganisms

    Directory of Open Access Journals (Sweden)

    Haruka Fukamachi

    2015-01-01

    Full Text Available Oral mucositis (OM in cancer patients induced by chemotherapy or radiotherapy has a significant impact on quality of life, and causes considerable morbidity. Oral microorganisms are likely to intensify the inflammatory process and aggravate the formation of ulcers. Hangeshashinto (HST, a Japanese kampo medicine, has been reported to be effective when used as a gargle for the treatment of OM. To clarify the effects of HST on oral microorganisms, we assessed its antimicrobial activity against 27 microbial species, including 19 oral bacteria and one fungus. HST extract inhibited the growth of Gram-negative bacteria, including Fusobacterium nucleatum, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, Prevotella melaninogenica, Tannerella forsythia, Treponema denticola, and Porphyromonas asaccharolytica, though inhibitory effects were less pronounced for Gram-positive bacteria and the fungal strain. We then investigated the effects of antibacterial activities on 15 purified ingredients of HST and determined that baicalein, berberine, coptisine, [6]-shogaol, and homogentisic acid actively inhibited the growth of these bacteria. These findings showed that HST inhibits the growth of specific Gram-negative periodontopathogenic bacteria, which are significant pathogens in OM, without disturbing the normal oral flora. Our data suggest that HST may be a useful treatment for OM in patients undergoing anticancer treatment.

  13. MALDI-TOF MS identification of anaerobic bacteria: assessment of pre-analytical variables and specimen preparation techniques.

    Science.gov (United States)

    Hsu, Yen-Michael S; Burnham, Carey-Ann D

    2014-06-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has emerged as a tool for identifying clinically relevant anaerobes. We evaluated the analytical performance characteristics of the Bruker Microflex with Biotyper 3.0 software system for identification of anaerobes and examined the impact of direct formic acid (FA) treatment and other pre-analytical factors on MALDI-TOF MS performance. A collection of 101 anaerobic bacteria were evaluated, including Clostridium spp., Propionibacterium spp., Fusobacterium spp., Bacteroides spp., and other anaerobic bacterial of clinical relevance. The results of our study indicate that an on-target extraction with 100% FA improves the rate of accurate identification without introducing misidentification (Panaerobes grown in suboptimal conditions, such as on selective culture media and following oxygen exposure. In conclusion, we report on a number of simple and cost-effective pre- and post-analytical modifications could enhance MALDI-TOF MS identification for anaerobic bacteria. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Stress hormone epinephrine (adrenaline) and norepinephrine (noradrenaline) effects on the anaerobic bacteria.

    Science.gov (United States)

    Boyanova, Lyudmila

    2017-04-01

    Microbial endocrinology is a relatively new research area that already encompasses the anaerobes. Stress hormones, epinephrine and norepinephrine, can affect the growth of anaerobic bacteria such as Fusobacterium nucleatum, Prevotella spp., Porhyromonas spp., Tanerella forsythia and Propionibacterium acnes and can increase virulence gene expression, iron acquisition and many virulence factors of some anaerobic species such as Clostridium perfringens, Porphyromonas gingivalis and Brachyspira pilosicoli. Epinephrine and norepinephrine effects can lead to a growth increase or decrease, or no effect on the growth of the anaerobes. The effects are species-specific and perhaps strain-specific. Discrepancies in the results of some studies can be due to the different methods and media used, catecholamine concentrations, measurement techniques and the low number of strains tested. Biological effects of the stress hormones on the anaerobes may range from halitosis and a worsening of periodontal diseases to tissue damages and atherosclerotic plaque ruptures. Optimizations of the research methods and a detailed assessment of the catecholamine effects in conditions mimicking those in affected organs and tissues, as well as the effects on the quorum sensing and virulence of the anaerobes and the full spectrum of biological consequences of the effects are interesting topics for further evaluation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Occurrence of periodontal pathogens in ethnic groups from a native Brazilian reservation.

    Science.gov (United States)

    Gaetti-Jardim, Elerson; Pereira, Maurício Fabiano; Vieira, Evanice Menezes Marçal; Schweitzer, Christiane Marie; Okamoto, Ana Cláudia; Ávila-Campos, Mario J

    2015-06-01

    The present study was designed to evaluate the occurrence of periodontal pathogens in the subgingival biofilm of 100 native Brazilians living at the Umutina Indian Reservation, Mato Grosso State, Brazil. Periodontal clinical examinations were carried out prior to collection of subgingival biofilm, and the presence of 14 periodontal microorganisms was evaluated by polymerase chain reaction (PCR). The prevalence and risk analysis was performed using Cochran and Mantel-Haenszel statistics for dichotomous variables or Pearson's chi-squared test for analysis of proportions when variables had three or more categories. The interrelations between clinical and microbiological parameters were assessed using Fisher's exact test and the Mann-Whitney U test. Individuals with chronic periodontitis were frequently colonized by the association between Porphyromonas gingivalis and Campylobacter rectus, P. gingivalis and Prevotella intermedia, or P. gingivalis and Tannerella forsythia. Patients with chronic periodontitis were also colonized by Porphyromonas gulae and P. intermedia or by the association between P. gulae and T. forsythia. P. gulae was detected only in the subgingival samples from natives on a traditional diet. Gingival bleeding was associated with Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, T. forsythia, P. gingivalis, P. gulae, Porphyromonas endodontalis, P. intermedia, and Prevotella nigrescens. Treponema denticola was uncommon. Peculiar microbiota was demonstrated to be associated with different periodontal disease statuses in native Brazilians, with modest occurrence of certain pathogens, such as T. denticola, and the presence of P. gulae in natives with gingivitis or chronic periodontitis. Copyright © 2015. Published by Elsevier Ltd.

  16. The microbiota of acute apical abscesses.

    Science.gov (United States)

    Siqueira, J F; Rôças, I N

    2009-01-01

    As the breadth of bacterial diversity in the oral cavity has been deciphered by molecular studies, several newly identified species/phylotypes have emerged as potential pathogens. We hypothesized that many of these species/phylotypes could also be involved with the etiology of endodontic abscesses. Abscess aspirates from 42 persons were analyzed for the presence of 81 species/phylotypes by means of a reverse-capture checkerboard hybridization assay. Associations between the most frequently detected taxa were calculated. The most prevalent taxa were Fusobacterium nucleatum, Parvimonas micra, and Porphyromonas endodontalis. Other frequently found taxa included Olsenella uli, streptococci, Eikenella corrodens, some as-yet-uncultivated phylotypes (Bacteroidetes clone X083 and Synergistes clone BA121), and newly named species (Prevotella baroniae and Dialister invisus). Several positive bacterial associations were disclosed. Findings not only strengthen the association of many cultivable species with abscesses, but also include some newly named species and uncultivated phylotypes in the set of candidate pathogens associated with this disease.

  17. Oral and dental infections with anaerobic bacteria: clinical features, predominant pathogens, and treatment.

    Science.gov (United States)

    Tanner, A; Stillman, N

    1993-06-01

    Microbial populations colonizing the teeth are a major source of pathogens responsible for oral and dental infections, including periodontal diseases, gingivitis, pericoronitis, endodontitis, peri-implantitis, and postextraction infections. Each entity has distinct clinical and microbial features. Bacterial species associated with oral infections include Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, Campylobacter rectus, Eubacterium species, Fusobacterium nucleatum, Eikenella corrodens, and Peptostreptococcus micros. Treponema pallidum-related spirochetes have been associated with acute necrotizing ulcerative gingivitis. Porphyromonas endodontalis appears to be specifically related to endodontic infections. Oral infections in medically compromised patients, including those with AIDS, are associated with similar species and are usually complicated by superinfection with enteric and Candida species. Isolation of species causing oral infections requires the collection of appropriate samples and the use of strictly anaerobic techniques. Rapid selective culture, immunofluorescence, and DNA probe methods have been developed for the identification of these oral species. The varied measures required in the management of oral and dental infections may include antimicrobial therapy. Accurate microbiological diagnosis, including antibiotic susceptibility testing, is indicated for cases that do not respond to therapy.

  18. The role of lipopolysaccharide in infectious bone resorption of periapical lesion.

    Science.gov (United States)

    Hong, Chi-Yuan; Lin, Sze-Kwan; Kok, Sang-Heng; Cheng, Shih-Jung; Lee, Ming-Shu; Wang, Tong-Mei; Chen, Chuan-Shuo; Lin, Li-Deh; Wang, Juo-Song

    2004-03-01

    The role of lipopolysaccharide (LPS) in periapical lesion-induced bone resorption was investigated. Polymyxin B (PMB), a specific inhibitor of LPS, was evaluated to treat the apical lesion. Lipopolysaccharide isolated from two common endodontic pathogens, Fusobacterium nucleatum and Porphyromonas endodontalis, stimulated mouse macrophage (J774) to release interleukin-1alpha (IL-1 alpha) and tumor necrosis factor-alpha (TNF-alpha) in a time-dependent manner. Combination of LPS further enhanced the stimulation. PMB inhibited these effects significantly. LPS also stimulated matrix metalloproteinase-1 (MMP-1) gene expression in J774, whereas anti-IL-1 alpha and anti-TNF-alpha antibodies, as well as PMB, diminished this effect. A disease model of periapical lesion was established in Wistar rat. Administration of PMB reduced the extent of lesion-associated bone resorption by 76% to approximately 80%, and simultaneously reduced the numbers of MMP-1-producing macrophages. It is suggested that LPS released from the infected root canal triggers the synthesis of IL-1 alpha and TNF-alpha from macrophages. These pro-inflammatory cytokines up-regulate the production of MMP-1 by macrophages to promote periapical bone resorption.

  19. Detection of hydrogen cyanide from oral anaerobes by cavity ring down spectroscopy

    Science.gov (United States)

    Chen, Wen; Roslund, Kajsa; Fogarty, Christopher L.; Pussinen, Pirkko J.; Halonen, Lauri; Groop, Per-Henrik; Metsälä, Markus; Lehto, Markku

    2016-03-01

    Hydrogen cyanide (HCN) has been recognized as a potential biomarker for non-invasive diagnosis of Pseudomonas aeruginosa infection in the lung. However, the oral cavity is a dominant production site for exhaled HCN and this contribution can mask the HCN generated in the lung. It is thus important to understand the sources of HCN production in the oral cavity. By screening of oral anaerobes for HCN production, we observed that the genus of Porphyromonas, Prevotella and Fusobacterium generated low levels of HCN in vitro. This is the first study to show that oral anaerobes are capable of producing HCN in vitro. Further investigations were conducted on the species of P. gingivalis and we successfully detected HCN production (0.9-10.9 ppb) in the headspace of three P. gingivalis reference strains (ATCC 33277, W50 and OMG 434) and one clinical isolate. From P. gingivalis ATCC 33277 and W50, a strong correlation between HCN and CO2 concentrations (rs = 0.89, p < 0.001) was observed, indicating that the HCN production of P. gingivalis might be connected with the bacterial metabolic activity. These results indicate that our setup could be widely applied to the screening of in vitro HCN production by both aerobic and anaerobic bacteria.

  20. Antibacterial activity of sphingoid bases and fatty acids against Gram-positive and Gram-negative bacteria.

    Science.gov (United States)

    Fischer, Carol L; Drake, David R; Dawson, Deborah V; Blanchette, Derek R; Brogden, Kim A; Wertz, Philip W

    2012-03-01

    There is growing evidence that the role of lipids in innate immunity is more important than previously realized. How lipids interact with bacteria to achieve a level of protection, however, is still poorly understood. To begin to address the mechanisms of antibacterial activity, we determined MICs and minimum bactericidal concentrations (MBCs) of lipids common to the skin and oral cavity--the sphingoid bases D-sphingosine, phytosphingosine, and dihydrosphingosine and the fatty acids sapienic acid and lauric acid--against four Gram-negative bacteria and seven Gram-positive bacteria. Exact Kruskal-Wallis tests of these values showed differences among lipid treatments (P 500 μg/ml). Sapienic acid (MBC range, 31.3 to 375.0 μg/ml) was active against Streptococcus sanguinis, Streptococcus mitis, and Fusobacterium nucleatum but not active against Escherichia coli, Staphylococcus aureus, S. marcescens, P. aeruginosa, Corynebacterium bovis, Corynebacterium striatum, and Corynebacterium jeikeium (MBC > 500 μg/ml). Lauric acid (MBC range, 6.8 to 375.0 μg/ml) was active against all bacteria except E. coli, S. marcescens, and P. aeruginosa (MBC > 500 μg/ml). Complete killing was achieved as early as 0.5 h for some lipids but took as long as 24 h for others. Hence, sphingoid bases and fatty acids have different antibacterial activities and may have potential for prophylactic or therapeutic intervention in infection.

  1. Molecular analysis of microbial diversity in advanced caries.

    Science.gov (United States)

    Chhour, Kim-Ly; Nadkarni, Mangala A; Byun, Roy; Martin, F Elizabeth; Jacques, Nicholas A; Hunter, Neil

    2005-02-01

    Real-time PCR analysis of the total bacterial load in advanced carious lesions has shown that the total load exceeds the number of cultivable bacteria. This suggests that an unresolved complexity exists in bacteria associated with advanced caries. In this report, the profile of the microflora of carious dentine was explored by using DNA extracted from 10 lesions selected on the basis of comparable total microbial load and on the relative abundance of Prevotella spp. Using universal primers for the 16S rRNA gene, PCR amplicons were cloned, and approximately 100 transformants were processed for each lesion. Phylogenetic analysis of 942 edited sequences demonstrated the presence of 75 species or phylotypes in the 10 carious lesions. Up to 31 taxa were represented in each sample. A diverse array of lactobacilli were found to comprise 50% of the species, with prevotellae also abundant, comprising 15% of the species. Other taxa present in a number of lesions or occurring with high abundance included Selenomonas spp., Dialister spp., Fusobacterium nucleatum, Eubacterium spp., members of the Lachnospiraceae family, Olsenella spp., Bifidobacterium spp., Propionibacterium sp., and Pseudoramibacter alactolyticus. The mechanisms by which such diverse patterns of bacteria extend carious lesions, including the aspect of infection of the vital dental pulp, remain unclear.

  2. Antimicrobial susceptibility of anaerobic bacteria in New Zealand: 1999-2003.

    Science.gov (United States)

    Roberts, Sally A; Shore, Keith P; Paviour, Susan D; Holland, David; Morris, Arthur J

    2006-05-01

    Routine susceptibility testing of all anaerobic organisms is not advocated, but it is useful for laboratories to test periodically for anaerobic organisms and provide local susceptibility data to guide therapy. This study reports the national trend of antibiotic susceptibility of clinically significant anaerobes in New Zealand. Clinical isolates were tested using standardized methods against a range of antibiotics commonly used to treat anaerobic infections. Susceptibility was determined using NCCLS criteria. The change in susceptibility trends between this study and earlier studies was measured by comparing the geometric mean of the MIC. A total of 364 anaerobes were tested. Penicillin had poor activity against Bacteroides spp., Prevotella spp., Eubacterium spp., Clostridium tertium and Veillonella spp. In general, Fusobacterium spp., Bacteroides ureolyticus, Propionibacterium spp., Clostridium perfringens and anaerobic streptococci isolates, with the exception of Peptostreptococcus anaerobius, were penicillin susceptible. Amoxicillin/clavulanate showed good activity against most anaerobes, but resistance was seen with Bacteroides fragilis group and P. anaerobius isolates. Cefoxitin was more active than cefotetan, particularly against non-B. fragilis species, Eubacterium spp. and P. anaerobius. Meropenem and imipenem showed good activity against all anaerobes, with only 2 and 4% of Bacteroides spp., respectively, showing resistance. With the exception of Propionibacterium acnes isolates, which are predictably resistant, metronidazole was active against all anaerobes tested. There has been little change in susceptibility since 1997. Metronidazole, cefoxitin, piperacillin/tazobactam and amoxicillin/clavulanate remain good empirical choices when anaerobes are expected in our setting. No clinically relevant changes in susceptibility over time were found.

  3. Associations of endodontic symptoms and signs with particular combinations of specific bacteria.

    Science.gov (United States)

    Gomes, B P; Lilley, J D; Drucker, D B

    1996-03-01

    Significant associations have been reported between (a) specific bacterial species isolated from root canals and (b) between individual bacterial species and endodontic symptoms and signs. The prime objective of this study was to determine whether particular combinations of specific bacteria are associated with individual endodontic symptoms and signs. Seventy root canals were investigated microbiologically taking care to maintain the viability of obligate anaerobes, which accounted for 64% of the total species isolated, including Peptostreptococcus micros, Prevotella melaninogenica, Prevotella oralis, Eubacterium aerofaciens, Eubacterium lentum, Fusobacterium nucleatum, Prevotella buccae and Prevotella intermedia. Significant associations were found between individual clinical features and the following pairs of species: (a) pain (37 cases) and Peptostreptococcus spp./Prevotella spp., Peptostreptococcus spp./Prevotella melaninogenica, Pstr. micros/Prev. melaninogenica (all P spp. (P spp./Eubacterium spp. (P spp./Eubacterium spp. (P < 0.05). Thus data from this investigation suggests that statistically significant associations exist between individual endodontic symptoms and signs and particular combinations of specific bacteria.

  4. Molecular methods for diagnosis of odontogenic infections.

    Science.gov (United States)

    Flynn, Thomas R; Paster, Bruce J; Stokes, Lauren N; Susarla, Srinivas M; Shanti, Rabie M

    2012-08-01

    Historically, the identification of microorganisms has been limited to species that could be cultured in the microbiology laboratory. The purpose of the present study was to apply molecular techniques to identify microorganisms in orofacial odontogenic infections (OIs). Specimens were obtained from subjects with clinical evidence of OI. To identify the microorganisms involved, 16S rRNA sequencing methods were used on clinical specimens. The name and number of the clones of each species identified and the combinations of species present were recorded for each subject. Descriptive statistics were computed for the study variables. Specimens of pus or wound fluid were obtained from 9 subjects. A mean of 7.4 ± 3.7 (standard deviation) species per case were identified. The predominant species detected in the present study that have previously been associated with OIs were Fusobacterium spp, Parvimonas micra, Porphyromonas endodontalis, and Prevotella oris. The predominant species detected in our study that have not been previously associated with OIs were Dialister pneumosintes and Eubacterium brachy. Unculturable phylotypes accounted for 24% of the species identified in our study. All species detected were obligate or facultative anaerobes. Streptococci were not detected. Molecular methods have enabled us to detect previously cultivated and not-yet-cultivated species in OIs; these methods could change our understanding of the pathogenic flora of orofacial OIs. Copyright © 2012 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

  5. Black-pigmented gram-negative anaerobes in endodontic infections.

    Science.gov (United States)

    Haapasalo, M

    1993-03-01

    Necrotic dental root canal infections are polymicrobial infections dominated by anaerobic bacteria. The number of different species in one canal is usually low, approx. 4-7 species. The species isolated most frequently belong to the genera Prevotella, Porphyromonas, Fusobacterium, Peptostreptococcus, Eubacterium and Streptococcus. The frequency of isolation of black-pigmented Gram-negative anaerobes in endodontic infections varies from 25% to > 50%. Pr. intermedia is the most commonly found pigmented species, followed by Pr. denticola and two Porphyromonas species, P. gingivalis and P. endodontalis. Several studies have shown that P. gingivalis and P. endodontalis are closely related to the presence of acute symptoms in endodontic infections, whereas other black-pigmented Gram-negative anaerobes are not. However, several other species may also be involved in acute infections. Moreover, Porphyromonas species have occasionally been isolated from cases with no symptoms. Although Porphyromonas spp. are clearly related to symptoms at the beginning of therapy, they are not important for the prognosis of the treatment.

  6. Competition between yogurt probiotics and periodontal pathogens in vitro.

    Science.gov (United States)

    Zhu, Yunwo; Xiao, Liying; Shen, Da; Hao, Yuqing

    2010-09-01

    To investigate the competition between probiotics in bio-yogurt and periodontal pathogens in vitro. The antimicrobial activity of bio-yogurt was studied by agar diffusion assays, using eight species of putative periodontal pathogens and a 'protective bacteria' as indicator strains. Four probiotic bacterial species (Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium) were isolated from yogurt and used to rate the competitive exclusion between probiotics and periodontal pathogens. Fresh yogurt inhibited all the periodontal pathogens included in this work, showing inhibition zones ranging from 9.3 (standard deviation 0.6) mm to 17.3 (standard deviation 1.7) mm, whereas heat-treated yogurt showed lower antimicrobial activity. In addition, neither fresh yogurt nor heat-treated yogurt inhibited the 'protective bacteria', Streptococcus sanguinis. The competition between yogurt probiotics and periodontal pathogens depended on the sequence of inoculation. When probiotics were inoculated first, Bifidobacterium inhibited Porphyromonas gingivalis, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Porphyromonas circumdentaria, and Prevotella nigrescens; L. acidophilus inhibited P. gingivalis, A. actinomycetemcomitans, P. circumdentaria, P. nigrescens, and Peptostreptococcus anaerobius; L. bulgaricus inhibited P. gingivalis, A. actinomycetemcomitans, and P. nigrescens; and S. thermophilus inhibited P. gingivalis, F. nucleatum, and P. nigrescens. However, their antimicrobial properties were reduced when both species (probiotics and periodontal pathogens) were inoculated simultaneously. When periodontal pathogens were inoculated first, Prevotella intermedia inhibited Bifidobacterium and S. thermophilus. The results demonstrated that bio-yogurt and the probiotics that it contains are capable of inhibiting specific periodontal pathogens but have no effect on the periodontal protective bacteria.

  7. Microbiologic picture of microflora offemale urogenital organs

    Directory of Open Access Journals (Sweden)

    Dyudyun A.D.

    2016-03-01

    Full Text Available In this report authors give a detailed analysis of microorganisms that can contaminate mucous membranes of female urogenital system in normal range and in the formation of certain dysbiotic position. The composition of microorganisms that colonize the mucous membrane of the urogenital tract depends on many exogenous and endogenous factors. The main microorganisms that provide colonizing resistance of vaginal biotop (VB are lactobacilli and lactobacteria. The protective properties of lactobacillus are implemented by antagonistic activity and ability to produce lysozyme and hydrogen peroxide. L. acidophilus, L. spp.; Propionbacterium spp.; Fusobacterium spp.; Porphyromonas spp.; Prevotella spp.; S. epidermidis and S. Saprophyticus novobionrezistentni and Streptococcus spp. and others are of definite importance in supporting normobiots of mucous membranes of urogenital system of women. Decreasing number of lactobacilli and other microorganisms of normobiota leads to colonization of mucous membranes of the vagina with G.vaginalis, B.bivies, B.disiens, B.melaninigenius, Mobiluncus, E.coli, E.fecalis, S.epidermidis and development of clinical manifestation of VB. The authors show microscopic and bacteriological characteristics of microorganisms that form normal microbiota and pathological states. The development of basic subjects promotes increasing number of the known microorganisms, important in the development of VB.

  8. Two mechanisms of oral malodor inhibition by zinc ions.

    Science.gov (United States)

    Suzuki, Nao; Nakano, Yoshio; Watanabe, Takeshi; Yoneda, Masahiro; Hirofuji, Takao; Hanioka, Takashi

    2018-01-18

    The aim of this study was to reveal the mechanisms by which zinc ions inhibit oral malodor. The direct binding of zinc ions to gaseous hydrogen sulfide (H2S) was assessed in comparison with other metal ions. Nine metal chlorides and six metal acetates were examined. To understand the strength of H2S volatilization inhibition, the minimum concentration needed to inhibit H2S volatilization was determined using serial dilution methods. Subsequently, the inhibitory activities of zinc ions on the growth of six oral bacterial strains related to volatile sulfur compound (VSC) production and three strains not related to VSC production were evaluated. Aqueous solutions of ZnCl2, CdCl2, CuCl2, (CH3COO)2Zn, (CH3COO)2Cd, (CH3COO)2Cu, and CH3COOAg inhibited H2S volatilization almost entirely. The strengths of H2S volatilization inhibition were in the order Ag+ > Cd2+ > Cu2+ > Zn2+. The effect of zinc ions on the growth of oral bacteria was strain-dependent. Fusobacterium nucleatum ATCC 25586 was the most sensitive, as it was suppressed by medium containing 0.001% zinc ions. Zinc ions have an inhibitory effect on oral malodor involving the two mechanisms of direct binding with gaseous H2S and suppressing the growth of VSC-producing oral bacteria.

  9. Effects of Protease Addition and Replacement of Soybean Meal by Corn Gluten Meal on the Growth of Broilers and on the Environmental Performances of a Broiler Production System in Greece

    Science.gov (United States)

    Bonos, Eleftherios; Anestis, Vasileios; Filioussis, Georgios; Papanastasiou, Dimitrios K.; Bartzanas, Thomas; Papaioannou, Nikolaos; Tzora, Athina; Skoufos, Ioannis

    2017-01-01

    An experimental study was conducted to examine the combined effects of adding a dietary protease, reducing the levels of soybean meal (SBM) and introducing corn gluten meal (CGM) in the ration of a group of broilers reared on a commercial Greek farm. Five hundred forty chicks were divided into three dietary treatments with six replicates of thirty birds each. The first group (Control) was fed a conventional diet based on corn and soybean meal, containing 21% w/w crude protein (CP). The second group (Soy-Prot) was supplied a corn and SBM-based diet containing a lower level of CP (20% w/w) and 200 mg of the protease RONOZYME® Proact per kg of feed. The third group (Gluten-Prot) was fed a diet without soybean-related constituents which was based on corn and CGM and with CP and protease contents identical to those of the diet of the Soy-Prot group. Body weight, feed intake, feed conversion ratio (FCR), intestinal microbiota populations and morphology, meat quality and cost were evaluated. Furthermore, a partial life cycle assessment (LCA) was performed in order to assess the potential environmental performance of the systems defined by these three dietary treatments and identify their environmental hot-spots. The growth performance of the broilers supplied the Soy-Prot diet was similar to the broilers supplied the Control diet. However, the broilers which were fed the Gluten-Prot diet at the end of the trial showed a tendency (P≤0.010) for lower weight gain and feed intake compared to those of the Control diet. When compared to the Control group, lower counts of C. perfringens (P≤0.05) were detected in the ileum and cecum parts, and lower counts of F. necrophorum (P≤0.001) were detected in the cecum part of the birds from the Gluten-Prot group. The evaluation of intestinal morphometry showed that the villus height and crypt depth values were not significantly different (P>0.05) among the experimental groups for the duodenum, jejunum and ileum parts. No

  10. Effects of Protease Addition and Replacement of Soybean Meal by Corn Gluten Meal on the Growth of Broilers and on the Environmental Performances of a Broiler Production System in Greece.

    Directory of Open Access Journals (Sweden)

    Ilias Giannenas

    Full Text Available An experimental study was conducted to examine the combined effects of adding a dietary protease, reducing the levels of soybean meal (SBM and introducing corn gluten meal (CGM in the ration of a group of broilers reared on a commercial Greek farm. Five hundred forty chicks were divided into three dietary treatments with six replicates of thirty birds each. The first group (Control was fed a conventional diet based on corn and soybean meal, containing 21% w/w crude protein (CP. The second group (Soy-Prot was supplied a corn and SBM-based diet containing a lower level of CP (20% w/w and 200 mg of the protease RONOZYME® Proact per kg of feed. The third group (Gluten-Prot was fed a diet without soybean-related constituents which was based on corn and CGM and with CP and protease contents identical to those of the diet of the Soy-Prot group. Body weight, feed intake, feed conversion ratio (FCR, intestinal microbiota populations and morphology, meat quality and cost were evaluated. Furthermore, a partial life cycle assessment (LCA was performed in order to assess the potential environmental performance of the systems defined by these three dietary treatments and identify their environmental hot-spots. The growth performance of the broilers supplied the Soy-Prot diet was similar to the broilers supplied the Control diet. However, the broilers which were fed the Gluten-Prot diet at the end of the trial showed a tendency (P≤0.010 for lower weight gain and feed intake compared to those of the Control diet. When compared to the Control group, lower counts of C. perfringens (P≤0.05 were detected in the ileum and cecum parts, and lower counts of F. necrophorum (P≤0.001 were detected in the cecum part of the birds from the Gluten-Prot group. The evaluation of intestinal morphometry showed that the villus height and crypt depth values were not significantly different (P>0.05 among the experimental groups for the duodenum, jejunum and ileum parts. No

  11. Effects of Protease Addition and Replacement of Soybean Meal by Corn Gluten Meal on the Growth of Broilers and on the Environmental Performances of a Broiler Production System in Greece.

    Science.gov (United States)

    Giannenas, Ilias; Bonos, Eleftherios; Anestis, Vasileios; Filioussis, Georgios; Papanastasiou, Dimitrios K; Bartzanas, Thomas; Papaioannou, Nikolaos; Tzora, Athina; Skoufos, Ioannis

    2017-01-01

    An experimental study was conducted to examine the combined effects of adding a dietary protease, reducing the levels of soybean meal (SBM) and introducing corn gluten meal (CGM) in the ration of a group of broilers reared on a commercial Greek farm. Five hundred forty chicks were divided into three dietary treatments with six replicates of thirty birds each. The first group (Control) was fed a conventional diet based on corn and soybean meal, containing 21% w/w crude protein (CP). The second group (Soy-Prot) was supplied a corn and SBM-based diet containing a lower level of CP (20% w/w) and 200 mg of the protease RONOZYME® Proact per kg of feed. The third group (Gluten-Prot) was fed a diet without soybean-related constituents which was based on corn and CGM and with CP and protease contents identical to those of the diet of the Soy-Prot group. Body weight, feed intake, feed conversion ratio (FCR), intestinal microbiota populations and morphology, meat quality and cost were evaluated. Furthermore, a partial life cycle assessment (LCA) was performed in order to assess the potential environmental performance of the systems defined by these three dietary treatments and identify their environmental hot-spots. The growth performance of the broilers supplied the Soy-Prot diet was similar to the broilers supplied the Control diet. However, the broilers which were fed the Gluten-Prot diet at the end of the trial showed a tendency (P≤0.010) for lower weight gain and feed intake compared to those of the Control diet. When compared to the Control group, lower counts of C. perfringens (P≤0.05) were detected in the ileum and cecum parts, and lower counts of F. necrophorum (P≤0.001) were detected in the cecum part of the birds from the Gluten-Prot group. The evaluation of intestinal morphometry showed that the villus height and crypt depth values were not significantly different (P>0.05) among the experimental groups for the duodenum, jejunum and ileum parts. No

  12. Composition and Antibacterial Activity of the Essential Oils of Orthosiphon stamineus Benth and Ficus deltoidea Jack against Pathogenic Oral Bacteria

    Directory of Open Access Journals (Sweden)

    Nuramirah Azizan

    2017-12-01

    Full Text Available In this study, the essential oils of Orthosiphon stamineus Benth and Ficus deltoidea Jack were evaluated for their antibacterial activity against invasive oral pathogens, namely Enterococcus faecalis, Streptococcus mutans, Streptococcus mitis, Streptococcus salivarius, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Fusobacterium nucleatum. Chemical composition of the oils was analyzed using gas chromatography (GC and gas chromatography-mass spectrometry (GC-MS. The antibacterial activity of the oils and their major constituents were investigated using the broth microdilution method (minimal inhibitory concentration (MIC and minimal bactericidal concentration (MBC. Susceptibility test, anti-adhesion, anti-biofilm, checkerboard and time-kill assays were also carried out. Physiological changes of the bacterial cells after exposure to the oils were observed under the field emission scanning electron microscope (FESEM. O. stamineus and F. deltoidea oils mainly consisted of sesquiterpenoids (44.6% and 60.9%, respectively, and β-caryophyllene was the most abundant compound in both oils (26.3% and 36.3%, respectively. Other compounds present in O. stamineus were α-humulene (5.1% and eugenol (8.1%, while α-humulene (5.5% and germacrene D (7.7% were dominant in F. deltoidea. The oils of both plants showed moderate to strong inhibition against all tested bacteria with MIC and MBC values ranging 0.63–2.5 mg/mL. However, none showed any inhibition on monospecies biofilms. The time-kill assay showed that combination of both oils with amoxicillin at concentrations of 1× and 2× MIC values demonstrated additive antibacterial effect. The FESEM study showed that both oils produced significant alterations on the cells of Gram-negative bacteria as they became pleomorphic and lysed. In conclusion, the study indicated that the oils of O. stamineus and F. deltoidea possessed moderate to strong antibacterial properties against the seven

  13. Composition and Antibacterial Activity of the Essential Oils of Orthosiphon stamineus Benth and Ficus deltoidea Jack against Pathogenic Oral Bacteria.

    Science.gov (United States)

    Azizan, Nuramirah; Mohd Said, Shahida; Zainal Abidin, Zamirah; Jantan, Ibrahim

    2017-12-05

    In this study, the essential oils of Orthosiphon stamineus Benth and Ficus deltoidea Jack were evaluated for their antibacterial activity against invasive oral pathogens, namely Enterococcus faecalis , Streptococcus mutans , Streptococcus mitis , Streptococcus salivarius , Aggregatibacter actinomycetemcomitans , Porphyromonas gingivalis and Fusobacterium nucleatum . Chemical composition of the oils was analyzed using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The antibacterial activity of the oils and their major constituents were investigated using the broth microdilution method (minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC)). Susceptibility test, anti-adhesion, anti-biofilm, checkerboard and time-kill assays were also carried out. Physiological changes of the bacterial cells after exposure to the oils were observed under the field emission scanning electron microscope (FESEM). O. stamineus and F. deltoidea oils mainly consisted of sesquiterpenoids (44.6% and 60.9%, respectively), and β-caryophyllene was the most abundant compound in both oils (26.3% and 36.3%, respectively). Other compounds present in O. stamineus were α-humulene (5.1%) and eugenol (8.1%), while α-humulene (5.5%) and germacrene D (7.7%) were dominant in F. deltoidea . The oils of both plants showed moderate to strong inhibition against all tested bacteria with MIC and MBC values ranging 0.63-2.5 mg/mL. However, none showed any inhibition on monospecies biofilms. The time-kill assay showed that combination of both oils with amoxicillin at concentrations of 1× and 2× MIC values demonstrated additive antibacterial effect. The FESEM study showed that both oils produced significant alterations on the cells of Gram-negative bacteria as they became pleomorphic and lysed. In conclusion, the study indicated that the oils of O. stamineus and F. deltoidea possessed moderate to strong antibacterial properties against the seven strains

  14. Resistance profiles to antimicrobial agents in bacteria isolated from acute endodontic infections: systematic review and meta-analysis.

    Science.gov (United States)

    Lang, Pauline M; Jacinto, Rogério C; Dal Pizzol, Tatiane S; Ferreira, Maria Beatriz C; Montagner, Francisco

    2016-11-01

    Infected root canal or acute apical abscess exudates can harbour several species, including Fusobacterium, Porphyromonas, Prevotella, Parvimonas, Streptococcus, Treponema, Olsenella and not-yet cultivable species. A systematic review and meta-analysis was performed to assess resistance rates to antimicrobial agents in clinical studies that isolated bacteria from acute endodontic infections. Electronic databases and the grey literature were searched up to May 2015. Clinical studies in humans evaluating the antimicrobial resistance of primary acute endodontic infection isolates were included. PRISMA guidelines were followed. A random-effect meta-analysis was employed. The outcome was described as the pooled resistance rates for each antimicrobial agent. Heterogeneity and sensitivity analyses were performed. Subgroup analyses were conducted based upon report or not of the use of antibiotics prior to sampling as an exclusion factor (subgroups A and B, respectively). Data from seven studies were extracted. Resistance rates for 15 different antimicrobial agents were evaluated (range, 3.5-40.0%). Lower resistance rates were observed for amoxicillin/clavulanic acid and amoxicillin; higher resistance rates were detected for tetracycline. Resistance rates varied according to previous use of an antimicrobial agent as demonstrated by the subgroup analyses. Heterogeneity was observed for the resistance profiles of penicillin G in subgroup A and for amoxicillin, clindamycin, metronidazole and tetracycline in subgroup B. Sensitivity analyses demonstrated that resistance rates changed for metronidazole, clindamycin, tetracycline and amoxicillin. These findings suggest that clinical isolates had low resistance to β-lactams. Further well-designed studies are needed to clarify whether the differences in susceptibility among the antimicrobial agents may influence clinical responses to treatment. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights

  15. Biofilm formation on titanium alloy and anatase-Bactercline® coated titanium healing screws: an in vivo human study

    Directory of Open Access Journals (Sweden)

    Antonio Scarano

    2013-03-01

    Full Text Available Aim Bacterial adherence to implants is considered to be an important event in the pathogenesis of bacterial infections. In fact, this infection process is a first stage of peri-implant mucositis and peri-implantitis, and a positive correlation has been found between oral hygiene and marginal bone loss around implants in the edentulous mandible. Surface properties of transgingival implant components are important determinants in bacterial adhesion. The purpose of this study was to characterize the biofilm formation, in vivo, on healing screws made of titanium alloy or coated with a combination of anatase and Bactercline® product. Materials and methods Twenty-five patients, between 21- 37 years, in excellent systemic health, participated in this study. In each of the 25 participants, one anatase-Bactercline® coated healing screw (Test and one titanium alloy (TI6Al4V healing screw (Control were adapted to two different implants. Quantitative and qualitative biofilm formation on healing abutments was analyzed by culture method.Results Bacterial adherence to the two different healing screws used in this study were compared. Statistically significant differences were found between the Control and the Test group for both aerobic and anaerobic bacterial counts (p<0,05. The microflora consisted both of Gram-positive and Gram-negative bacteria, and displayed a high variability. The anaerobic S. intermedius, potentially “pathogenic”, was isolated only from the Control group. Both healing screws harbored primarily Gram-positive rods as Actinomyces spp, A. naeslundii, A. viscosus and the Gram-negative rods (Fusobacterium spp, Prevotella spp, Capnocythophaga spp were mostly found on the Control healing screws.Conclusion Anatase-Bactercline® coated healing screws reduce the number of initially adhering bacteria, formed mainly of Gram-positive microorgnisms, while, on the contrary, the microflora covering the titanium alloy healing screws was, for the

  16. Detección y prevalencia de patógenos periodontales de una población con periodontitis crónica en Uruguay mediante metodología convencional y metagenómica

    Directory of Open Access Journals (Sweden)

    Virginia Papone

    Full Text Available Resumen Las enfermedades periodontales son un significativo problema mundial a nivel de salud humana. Décadas de investigaciones, evidencian que en la mayoría de los casos la periodontitis crónica es la más común, caracterizada por ser de evolución lenta, con formación de bolsas periodontales, posterior reabsorción del hueso alveolar, pérdida y destrucción de piezas dentarias y tejido óseo. Si bien se reconoce el origen multifactorial en el desarrollo de la periodontitis, es relevante la participación de la microbiota subgingival en la etiología de la enfermedad periodontal. Algunas de las especies bacterianas patógenas que han sido asociadas con el desarrollo de la enfermedad periodontal son Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum, entre otras. En este estudio, nos propusimos investigar cuáles de éstas cinco especies estaban presentes en las bolsas periodontales de 51 pacientes uruguayos con periodontitis crónica. Para alcanzar éste objetivo se utilizó una técnica convencional microbiológica y metagenómica (multiplex-PCR. Los resultados de la técnica convencional microbiológica evidenciaron la presencia de A. actinomycetemcomitans (33% y de bacterias negras pigmentadas anaerobias (100% en las muestras. De los resultados obtenidos en la multiplex-PCR, se demostró que las especies de mayor prevalencia fueron F. nucleatum (100%, T. forsythia (92% y P. gingivalis (88%. Por el contrario, las especies de menor prevalencia fueron P. intermedia (39% y A. actinomycetemcomitans (33%

  17. The periodontal abscess (I). Clinical and microbiological findings.

    Science.gov (United States)

    Herrera, D; Roldán, S; González, I; Sanz, M

    2000-06-01

    Little information is available regarding the diagnosis and microbiology of periodontal abscesses. The aim of this descriptive clinical and microbiological study was to provide more information in order to help in the characterisation of the periodontal abscess associated to periodontitis. 29 consecutive patients with a periodontal abscess were studied by the assessment of clinical variables, including both subjective (pain, edema, redness and swelling) and objective (bleeding on probing, suppuration, probing pocket depth, tooth mobility and cervical lymphadenopathy) parameters. Microbiological samples were taken for anaerobic microbiology and processed by means of culture. Systemic involvement was also studied through the analysis of blood and urine samples using conventional laboratory standards. 62% of the abscesses affected untreated periodontitis patients, and 69% were associated with a molar tooth. More than 75% of the abscesses had moderate-severe scores related to edema, redness and swelling, and 90% of the patients reported pain. Bleeding occurred in all abscesses, while suppuration on sampling was detected in 66%. Mean associated pocket depth was 7.28 mm, and 79% of teeth presented some degree of mobility. Cervical lymphadenopathy was seen in 10% of patients, while elevated leucocyte counts were observed in 31.6%. The absolute number of neutrophils was elevated in 42% of the patients. High prevalences of putative periodontal pathogens were found, including Fusobacterium nucleatum, Peptostreptococcus micros, Porphyromonas gingivalis, Prevotella intermedia and Bacteroides forsythus. The periodontal abscess has clear clinical characteristics and is usually associated with severe periodontal destruction. This condition may cause systemic involvement and the lesion generally has a large bacterial mass with a high prevalence of well-recognised periodontal pathogens.

  18. Frequency and levels of candidate endodontic pathogens in acute apical abscesses as compared to asymptomatic apical periodontitis

    Science.gov (United States)

    Rôças, Isabela N.

    2018-01-01

    Introduction Acute apical abscess is caused by bacteria that leave the infected dental root canal to invade the periodontal tissues. Most species occurring in abscesses are also found in asymptomatic infections; therefore, the possibility exists that not only the presence of certain species but also their specific counts influence the appearance of symptoms. This molecular study compared the frequency and levels of several candidate endodontic pathogens in teeth with acute apical abscesses and asymptomatic apical periodontitis. Methods Samples were taken from the root canals of teeth with asymptomatic apical periodontitis (n = 73) and by aspiration of purulent exudate from acute abscesses (n = 55). DNA was extracted from samples and bacterial identifications were performed by a closed-ended semi-quantitative reverse-capture checkerboard approach targeting 40 bacterial species/phylotypes. Results Bacterial DNA was detected in all cases. In abscesses, the most prevalent taxa were Fusobacterium nucleatum (60%), Porphyromonas endodontalis (53%), Parvimonas micra (51%), and Streptococcus species (45%). The most frequently detected taxa in asymptomatic teeth were P. endodontalis (63%), Dialister invisus (58%), Olsenella uli (56%), and F. nucleatum (51%). None of the targeted taxa were significantly associated with abscesses when only mere presence was evaluated (p>0.05). However, semi-quantitative data demonstrated that P. endodontalis, Prevotella baroniae, Treponema denticola and Streptococcus species were significantly more frequent at levels >105 in abscesses than in asymptomatic cases (pabscesses in terms of frequency. However, some taxa were significantly found in higher levels in abscesses. Presence of a potentially virulent pathogen in high counts may increase the collective pathogenicity of the bacterial community and give rise to symptoms. PMID:29293651

  19. Sputum microbiota in tuberculosis as revealed by 16S rRNA pyrosequencing.

    Directory of Open Access Journals (Sweden)

    Man Kit Cheung

    Full Text Available BACKGROUND: Tuberculosis (TB remains a global threat in the 21st century. Traditional studies of the disease are focused on the single pathogen Mycobacterium tuberculosis. Recent studies have revealed associations of some diseases with an imbalance in the microbial community. Characterization of the TB microbiota could allow a better understanding of the disease. METHODOLOGY/PRINCIPAL FINDINGS: Here, the sputum microbiota in TB infection was examined by using 16S rRNA pyrosequencing. A total of 829,873 high-quality sequencing reads were generated from 22 TB and 14 control sputum samples. Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Fusobacteria were the five major bacterial phyla recovered, which together composed over 98% of the microbial community. Proteobacteria and Bacteroidetes were more represented in the TB samples and Firmicutes was more predominant in the controls. Sixteen major bacterial genera were recovered. Streptococcus, Neisseria and Prevotella were the most predominant genera, which were dominated by several operational taxonomic units grouped at a 97% similarity level. Actinomyces, Fusobacterium, Leptotrichia, Prevotella, Streptococcus, and Veillonella were found in all TB samples, possibly representing the core genera in TB sputum microbiota. The less represented genera Mogibacterium, Moryella and Oribacterium were enriched statistically in the TB samples, while a genus belonging to the unclassified Lactobacillales was enriched in the controls. The diversity of microbiota was similar in the TB and control samples. CONCLUSIONS/SIGNIFICANCE: The composition and diversity of sputum microbiota in TB infection was characterized for the first time by using high-throughput pyrosequencing. It lays the framework for examination of potential roles played by the diverse microbiota in TB pathogenesis and progression, and could ultimately facilitate advances in TB treatment.

  20. Minimum inhibitory concentration of the plant extracts′ combinations against dental caries and plaque microorganisms: An in vitro study

    Directory of Open Access Journals (Sweden)

    B R Chandra Shekar

    2016-01-01

    Full Text Available Introduction: Oral health status has witnessed marked advances in many industrialized countries. However, dental caries is consistently increasing in developing countries, and periodontal diseases are among most common afflictions to humankind. Approach best suited for developing countries is to focus on the prevention with innovative strategies. Hence, evolution of novel, innovative strategies to prevent dental caries and periodontal diseases is need of hour. Objective: To determine minimum inhibitory concentration (MIC of combinations of Acacia nilotica, Murraya koenigii L. Sprengel, Eucalyptus hybrid, and Psidium guajava against dental caries and plaque microorganisms and to qualitatively identify various phytochemical constituents in individual plant extracts and their quadruple combinations. Materials and Methods: MIC of the combinations of A. nilotica, M. koenigii L. Sprengel, Eucalyptus hybrid, and P. guajava on Streptococcus mutans, Lactobacillus acidophilus (dental caries bacteria, Streptococcus sanguis, Streptococcus salivarius (primary plaque colonizers, Fusobacterium nucleatum (secondary plaque colonizer, and Porphyromonas gingivalis (tertiary plaque colonizer was determined using broth dilution method. Series of dilutions of quadruple combinations ranging from 0.05% to 1.5% were prepared. 100 μL of each serial dilution of quadruple combinations was added to each tube containing bacterial culture. The optical density was noted after incubation in each tube to estimate the MIC for each bacterium. Results: MIC of the polyherbal combinations on S. mutans, S. sanguis, S. salivarius, L. acidophilus, F. nucleatum, and P. gingivalis was found to be 0.25%, 0.05%, 0.05%, 0.1%, 0.25%, and 0.25%, respectively. Conclusion: The quadruple combinations of these four plant extracts could be considered in the evolution of an indigenous polyherbal mouth rinse as the formulation inhibited all the bacteria tested in the present study at low

  1. Plaque bacterial microbiome diversity in children younger than 30 months with or without caries prior to eruption of second primary molars.

    Directory of Open Access Journals (Sweden)

    He Xu

    Full Text Available OBJECTIVE: Our primary objective is to phylogenetically characterize the supragingival plaque bacterial microbiome of children prior to eruption of second primary molars by pyrosequencing method for studying etiology of early childhood caries. METHODS: Supragingival plaque samples were collected from 10 caries children and 9 caries-free children. Plaque DNA was extracted, used to generate DNA amplicons of the V1-V3 hypervariable region of the bacterial 16S rRNA gene, and subjected to 454-pyrosequencing. RESULTS: On average, over 22,000 sequences per sample were generated. High bacterial diversity was noted in the plaque of children with caries [170 operational taxonomical units (OTU at 3% divergence] and caries-free children (201 OTU at 3% divergence with no significant difference. A total of 8 phyla, 15 classes, 21 orders, 30 families, 41 genera and 99 species were represented. In addition, five predominant phyla (Firmicute, Fusobacteria, Proteobacteria, Bacteroidetes and Actinobacteria and seven genera (Leptotrichia, Streptococcus, Actinomyces, Prevotella, Porphyromonas, Neisseria, and Veillonella constituted a majority of contents of the total microbiota, independent of the presence or absence of caries. Principal Component Analysis (PCA presented that caries-related genera included Streptococcus and Veillonella; while Leptotrichia, Selenomonas, Fusobacterium, Capnocytophaga and Porphyromonas were more related to the caries-free samples. Neisseria and Prevotella presented approximately in between. In both groups, the degree of shared organism lineages (as defined by species-level OTUs among individual supragingival plaque microbiomes was minimal. CONCLUSION: Our study represented for the first time using pyrosequencing to elucidate and monitor supragingival plaque bacterial diversity at such young age with second primary molar unerrupted. Distinctions were revealed between caries and caries-free microbiomes in terms of microbial community

  2. Effects of tongue cleaning on bacterial flora in tongue coating and dental plaque: a crossover study.

    Science.gov (United States)

    Matsui, Miki; Chosa, Naoyuki; Shimoyama, Yu; Minami, Kentaro; Kimura, Shigenobu; Kishi, Mitsuo

    2014-01-14

    The effects of tongue cleaning on reconstruction of bacterial flora in dental plaque and tongue coating itself are obscure. We assessed changes in the amounts of total bacteria as well as Fusobacterium nucleatum in tongue coating and dental plaque specimens obtained with and without tongue cleaning. We conducted a randomized examiner-blind crossover study using 30 volunteers (average 23.7 ± 3.2 years old) without periodontitis. After dividing randomly into 2 groups, 1 group was instructed to clean the tongue, while the other did not. On days 1 (baseline), 3, and 10, tongue coating and dental plaque samples were collected after recording tongue coating score (Winkel tongue coating index: WTCI). After a washout period of 3 weeks, the same examinations were performed with the subjects allocated to the alternate group. Genomic DNA was purified from the samples and applied to SYBR® Green-based real-time PCR to quantify the amounts of total bacteria and F. nucleatum. After 3 days, the WTCI score recovered to baseline, though the amount of total bacteria in tongue coating was significantly lower as compared to the baseline. In plaque samples, the bacterial amounts on day 3 and 10 were significantly lower than the baseline with and without tongue cleaning. Principal component analysis showed that variations of bacterial amounts in the tongue coating and dental plaque samples were independent from each other. Furthermore, we found a strong association between amounts of total bacteria and F. nucleatum in specimens both. Tongue cleaning reduced the amount of bacteria in tongue coating. However, the cleaning had no obvious contribution to inhibit dental plaque formation. Furthermore, recovery of the total bacterial amount induced an increase in F. nucleatum in both tongue coating and dental plaque. Thus, it is recommended that tongue cleaning and tooth brushing should both be performed for promoting oral health.

  3. A high-throughput microfluidic dental plaque biofilm system to visualize and quantify the effect of antimicrobials

    Science.gov (United States)

    Nance, William C.; Dowd, Scot E.; Samarian, Derek; Chludzinski, Jeffrey; Delli, Joseph; Battista, John; Rickard, Alexander H.

    2013-01-01

    Objectives Few model systems are amenable to developing multi-species biofilms in parallel under environmentally germane conditions. This is a problem when evaluating the potential real-world effectiveness of antimicrobials in the laboratory. One such antimicrobial is cetylpyridinium chloride (CPC), which is used in numerous over-the-counter oral healthcare products. The aim of this work was to develop a high-throughput microfluidic system that is combined with a confocal laser scanning microscope (CLSM) to quantitatively evaluate the effectiveness of CPC against oral multi-species biofilms grown in human saliva. Methods Twenty-four-channel BioFlux microfluidic plates were inoculated with pooled human saliva and fed filter-sterilized saliva for 20 h at 37°C. The bacterial diversity of the biofilms was evaluated by bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP). The antimicrobial/anti-biofilm effect of CPC (0.5%–0.001% w/v) was examined using Live/Dead stain, CLSM and 3D imaging software. Results The analysis of biofilms by bTEFAP demonstrated that they contained genera typically found in human dental plaque. These included Aggregatibacter, Fusobacterium, Neisseria, Porphyromonas, Streptococcus and Veillonella. Using Live/Dead stain, clear gradations in killing were observed when the biofilms were treated with CPC between 0.5% and 0.001% w/v. At 0.5% (w/v) CPC, 90% of the total signal was from dead/damaged cells. Below this concentration range, less killing was observed. In the 0.5%–0.05% (w/v) range CPC penetration/killing was greatest and biofilm thickness was significantly reduced. Conclusions This work demonstrates the utility of a high-throughput microfluidic–CLSM system to grow multi-species oral biofilms, which are compositionally similar to naturally occurring biofilms, to assess the effectiveness of antimicrobials. PMID:23800904

  4. Effect of an oxygenating agent on oral bacteria in vitro and on dental plaque composition in healthy young adults.

    Science.gov (United States)

    Fernandez y Mostajo, Mercedes; van der Reijden, Wil A; Buijs, Mark J; Beertsen, Wouter; Van der Weijden, Fridus; Crielaard, Wim; Zaura, Egija

    2014-01-01

    Oral bacteria live in symbiosis with the host. Therefore, when mouthwashes are indicated, selective inhibition of taxa contributing to disease is preferred instead of broad-spectrum antimicrobials. The potential selectivity of an oxygenating mouthwash, Ardox-X® (AX), has not been assessed. The aim of this study was to determine the antimicrobial potential of AX and the effects of a twice-daily oral rinse on dental plaque composition. In vitro, 16 oral bacterial strains were tested using agar diffusion susceptibility, minimum inhibitory and minimum bactericidal concentration tests. A pilot clinical study was performed with 25 healthy volunteers. Clinical assessments and microbiological sampling of supragingival plaque were performed at 1 month before the experiment (Pre-exp), at the start of the experiment (Baseline) and after the one-week experimental period (Post-exp). During the experiment individuals used AX mouthwash twice daily in absence of other oral hygiene measures. The microbiological composition of plaque was assessed by 16S rRNA gene amplicon sequencing. AX showed high inter-species variation in microbial growth inhibition. The tested Prevotella strains and Fusobacterium nucleatum showed the highest sensitivity, while streptococci and Lactobacillus acidophilus were most resistant to AX. Plaque scores at Pre-exp and Baseline visits did not differ significantly (p = 0.193), nor did the microbial composition of plaque. During a period of 7-days non-brushing but twice daily rinsing plaque scores increased from 2.21 (0.31) at Baseline to 2.43 (0.39) Post-exp. A significant microbial shift in composition was observed: genus Streptococcus and Veillonella increased while Corynebacterium, Haemophilus, Leptotrichia, Cardiobacterium and Capnocytophaga decreased (p ≤ 0.001). AX has the potential for selective inhibition of oral bacteria. The shift in oral microbiome after 1 week of rinsing deserves further research.

  5. 24-hour evaluation of dental plaque bacteria and halitosis after consumption of a single placebo or dental treat by dogs.

    Science.gov (United States)

    Jeusette, Isabelle C; Román, Aurora Mateo; Torre, Celina; Crusafont, Josep; Sánchez, Nuria; Sánchez, Maria C; Pérez-Salcedo, Leire; Herrera, David

    2016-06-01

    OBJECTIVE To determine whether consumption of a single dental treat with specific mechanical properties and active ingredients would provide a 24-hour effect on dental plaque bacteria and halitosis in dogs. ANIMALS 10 dogs of various breeds from a privately owned colony that had received routine dental scaling and polishing 4 weeks before the study began. PROCEDURES Dogs were randomly assigned to receive 1 placebo or dental treat first. A 4-week washout period was provided, and then dogs received the opposite treatment. Oral plaque and breath samples were collected before and 0.5, 3, 12, and 24 hours after treat consumption. Volatile sulfur compounds (VSCs) concentration was measured in breath samples. Total aerobic, total anaerobic, Porphyromonas gulae, Prevotella intermedia-like, Tannerella forsythia, and Fusobacterium nucleatum bacterial counts (measured via bacterial culture) and total live bacterial counts, total live and dead bacterial counts, and bacterial vitality (measured via quantitative real-time PCR assay) were assessed in plaque samples. RESULTS Compared with placebo treat consumption, dental treat consumption resulted in a significant decrease in breath VSCs concentration and all plaque bacterial counts, without an effect on bacterial vitality. Effects of the dental treat versus the placebo treat persisted for 12 hours for several bacterial counts and for 24 hours for breath VSCs concentration. CONCLUSIONS AND CLINICAL RELEVANCE Although clinical benefits should be investigated in larger scale, longer-term studies, results of this study suggested that feeding the evaluated dental treat may help to decrease oral bacterial growth in dogs for 12 hours and oral malodor for 24 hours. A feeding interval of 12 hours is therefore recommended.

  6. Effect of an oxygenating agent on oral microorganisms in vitro and on dental plaque composition in healthy young adults

    Directory of Open Access Journals (Sweden)

    Mercedes eFernandez y Mostajo

    2014-07-01

    Full Text Available Oral bacteria live in symbiosis with the host. Therefore, when mouthwashes are indicated, selective inhibition of taxa contributing to disease is preferred instead of broad-spectrum antimicrobials. The potential selectivity of an oxygenating mouthwash, Ardox-X® (AX, has not been assessed. The aim of this study was to determine the antimicrobial potential of AX and the effects of a twice-daily oral rinse on dental plaque composition. Material and methods: In vitro, 16 oral bacterial strains were tested using agar diffusion susceptibility, minimum inhibitory and minimum bactericidal concentration tests. A pilot clinical study was performed with 25 healthy volunteers. Clinical assessments and microbiological sampling of supragingival plaque were performed at one month before the experiment (Pre-exp, at the start of the experiment (Baseline and after the one-week experimental period (Post-exp. During the experiment individuals used AX mouthwash twice daily in absence of other oral hygiene measures. The microbiological composition of plaque was assessed by 16S rRNA gene amplicon sequencing. Results: AX showed high inter-species variation in microbial growth inhibition. The tested Prevotella strains and Fusobacterium nucleatum showed the highest sensitivity, while streptococci and Lactobacillus acidophilus were most resistant to AX. Plaque scores at Pre-exp and Baseline visits did not differ significantly (p = 0.193, nor did the microbial composition of plaque during a period of 7-days non-brushing but twice daily rinsing. Plaque scores increased from 2.21 (0.31 at Baseline to 2.43 (0.39 Post-exp. A significant microbial shift in composition was observed: genus Streptococcus and Veillonella increased while Corynebacterium, Haemophilus, Leptotrichia, Cardiobacterium and Capnocytophaga decreased (p ≤ 0.001. Conclusion: AX has the potential for selective inhibition of oral bacteria. The shift in oral microbiome after one week of rinsing deserves

  7. Bacterial community development in experimental gingivitis.

    Science.gov (United States)

    Kistler, James O; Booth, Veronica; Bradshaw, David J; Wade, William G

    2013-01-01

    Current knowledge of the microbial composition of dental plaque in early gingivitis is based largely on microscopy and cultural methods, which do not provide a comprehensive description of oral microbial communities. This study used 454-pyrosequencing of the V1-V3 region of 16S rRNA genes (approximately 500 bp), and bacterial culture, to characterize the composition of plaque during the transition from periodontal health to gingivitis. A total of 20 healthy volunteers abstained from oral hygiene for two weeks, allowing plaque to accumulate and gingivitis to develop. Plaque samples were analyzed at baseline, and after one and two weeks. In addition, plaque samples from 20 chronic periodontitis patients were analyzed for cross-sectional comparison to the experimental gingivitis cohort. All of the healthy volunteers developed gingivitis after two weeks. Pyrosequencing yielded a final total of 344,267 sequences after filtering, with a mean length of 354 bases, that were clustered into an average of 299 species-level Operational Taxonomic Units (OTUs) per sample. Principal coordinates analysis (PCoA) plots revealed significant shifts in the bacterial community structure of plaque as gingivitis was induced, and community diversity increased significantly after two weeks. Changes in the relative abundance of OTUs during the transition from health to gingivitis were correlated to bleeding on probing (BoP) scores and resulted in the identification of new health- and gingivitis-associated taxa. Comparison of the healthy volunteers to the periodontitis patients also confirmed the association of a number of putative periodontal pathogens with chronic periodontitis. Taxa associated with gingivitis included Fusobacterium nucleatum subsp. polymorphum, Lachnospiraceae [G-2] sp. HOT100, Lautropia sp. HOTA94, and Prevotella oulorum, whilst Rothia dentocariosa was associated with periodontal health. Further study of these taxa is warranted and may lead to new therapeutic approaches

  8. Effects of lactoferrin and lactoperoxidase-containing food on the oral hygiene status of older individuals: A randomized, double blinded, placebo-controlled clinical trial.

    Science.gov (United States)

    Morita, Yu; Ishikawa, Kentaro; Nakano, Manabu; Wakabayashi, Hiroyuki; Yamauchi, Koji; Abe, Fumiaki; Ooka, Takafumi; Hironaka, Shouji

    2017-05-01

    Lactoferrin and lactoperoxidase have antimicrobial effects against oral pathogens. This randomized, double-blinded, placebo-controlled parallel group study tested the efficacy of a lactoferrin and lactoperoxidase-containing tablet (LF + LPO tablet) in improving the oral hygiene status of older individuals. A total of 46 participants (31 nursing home residents and 15 healthy older individuals) were randomly assigned to receive either lactoferrin and lactoperoxidase-containing tablets or placebo tablets, and were asked to suck on a tablet after every meal for 8 weeks. Oral and bacteriological assessments were carried out at baseline, 4 weeks and 8 weeks. A total of 47 participants (test group n = 20; mean age 80.4 ± 6.4 years; placebo group n = 17; mean age 85.9 ± 6.7 years) were included in the efficacy analysis. In the test group, the total number of bacteria in the tongue coating was significantly reduced at 4 and 8 weeks compared with that at baseline, and the number of Porphyromonas gingivalis and Fusobacterium nucleatum was significantly reduced at 8 weeks. The total number of bacteria and the number of P. gingivalis in the supragingival plaque were significantly reduced at 8 weeks. Furthermore, there was a significant difference in the change in the number of P. gingivalis in supragingival plaque at 8 weeks between the two groups. Lactoferrin and lactoperoxidase-containing tablet ingestion showed antibacterial effects on periodontal bacteria present in the tongue coating and supragingival plaque, indicating that long-term ingestion could improve the oral hygiene of older individuals. Geriatr Gerontol Int 2017; 17: 714-721. © 2016 Japan Geriatrics Society.

  9. Antibacterial photocatalytic activity of different crystalline TiO2 phases in oral multispecies biofilm.

    Science.gov (United States)

    Pantaroto, Heloisa N; Ricomini-Filho, Antonio P; Bertolini, Martinna M; Dias da Silva, José Humberto; Azevedo Neto, Nilton F; Sukotjo, Cortino; Rangel, Elidiane C; Barão, Valentim A R

    2018-07-01

    Titanium dioxide (TiO 2 ) incorporation in biomaterials is a promising technology due to its photocatalytic and antibacterial activities. However, the antibacterial potential of different TiO 2 crystalline structures on a multispecies oral biofilm remains unknown. We hypothesized that the different crystalline TiO 2 phases present different photocatalytic and antibacterial activities. Three crystalline TiO 2 films were deposited by magnetron sputtering on commercially pure titanium (cpTi), in order to obtain four groups: (1) machined cpTi (control); (2) A-TiO 2 (anatase); (3) M-TiO 2 (mixture of anatase and rutile); (4) R-TiO 2 (rutile). The morphology, crystalline phase, chemical composition, hardness, elastic modulus and surface free energy of the surfaces were evaluated. The photocatalytic potential was assessed by methylene blue degradation assay. The antibacterial activity was evaluated on relevant oral bacteria, by using a multispecies biofilm (Streptococcus sanguinis, Actinomyces naeslundii and Fusobacterium nucleatum) formed on the treated titanium surfaces (16.5h) followed by UV-A light exposure (1h) to generate reactive oxygen species production. All TiO 2 films presented around 300nm thickness and improved the hardness and elastic modulus of cpTi surfaces (p0.05 vs. control). This study brings new insights on the development of extra oral protocols for the photocatalytic activity of TiO 2 in oral biofilm-associated disease. Anatase and mixture-TiO 2 showed antibacterial activity on this oral bacterial biofilm, being promising surface coatings for dental implant components. Copyright © 2018 The Academy of Dental Materials. All rights reserved.

  10. Changes in salivary microbiota increase volatile sulfur compounds production in healthy male subjects with academic-related chronic stress.

    Directory of Open Access Journals (Sweden)

    Bruno Dias Nani

    Full Text Available To investigate the associations among salivary bacteria, oral emanations of volatile sulfur compounds, and academic-related chronic stress in healthy male subjects.Seventy-eight healthy male undergraduate dental students were classified as stressed or not by evaluation of burnout, a syndrome attributed to academic-related chronic stress. This evaluation was carried out using the Maslach Burnout Inventory-Student Survey questionnaire. Oral emanations of hydrogen sulfide, methyl mercaptan, and dimethyl sulfide were measured using an Oral Chroma™ portable gas chromatograph. The amounts in saliva of total bacteria and seven bacteria associated with halitosis were quantified by qPCR. The in vitro production of H2S by S. moorei and/or F. nucleatum was also measured with the Oral Chroma™ instrument.The stressed students group showed increased oral emanations of hydrogen sulfide and dimethyl sulfide, together with higher salivary Solobacterium moorei levels (p < 0.05, Mann Whitney test. There were moderate positive correlations between the following pairs of variables: Fusobacterium nucleatum and S. moorei; F. nucleatum and hydrogen sulfide; Tannerella forsythia and F. nucleatum; T. forsythia and S. moorei. These correlations only occurred for the stressed group (p < 0.05, Spearman correlation. The in vitro experiment demonstrated that S. moorei increased H2S production by F. nucleatum (p < 0.05, ANOVA and Tukey's test.The increased amount of S. moorei in saliva, and its coexistence with F. nucleatum and T. forsythia, seemed to be responsible for increased oral hydrogen sulfide in the healthy male stressed subjects.

  11. Microbial transformation from normal oral microbiota to acute endodontic infections

    Directory of Open Access Journals (Sweden)

    Hsiao William WL

    2012-07-01

    Full Text Available Abstract Background Endodontic infections are a leading cause of oro-facial pain and tooth loss in western countries, and may lead to severe life-threatening infections. These infections are polymicrobial with high bacterial diversity. Understanding the spatial transition of microbiota from normal oral cavities through the infected root canal to the acute periapical abscess can improve our knowledge of the pathogenesis of endodontic infections and lead to more effective treatment. We obtained samples from the oral cavity, infected root canal and periapical abscess of 8 patients (5 with localized and 3 with systemic infections. Microbial populations in these samples were analyzed using next-generation sequencing of 16S rRNA amplicons. Bioinformatics tools and statistical tests with rigorous criteria were used to elucidate the spatial transition of the microbiota from normal to diseased sites. Results On average, 10,000 partial 16S rRNA gene sequences were obtained from each sample. All sequences fell into 11 different bacterial phyla. The microbial diversity in root canal and abscess samples was significantly lower than in the oral samples. Streptococcus was the most abundant genus in oral cavities while Prevotella and Fusobacterium were most abundant in diseased samples. The microbiota community structures of root canal and abscess samples were, however, more similar to each other than to the oral cavity microbiota. Using rigorous criteria and novel bioinformatics tools, we found that Granulicatella adiacens, Eubacterium yurii, Prevotella melaninogenica, Prevotella salivae, Streptococcus mitis, and Atopobium rimae were over-represented in diseased samples. Conclusions We used a novel approach and high-throughput methodologies to characterize the microbiota associated normal and diseased oral sites in the same individuals.

  12. Traditional Medicinal Plant Extracts and Natural Products with Activity against Oral Bacteria: Potential Application in the Prevention and Treatment of Oral Diseases

    Directory of Open Access Journals (Sweden)

    Enzo A. Palombo

    2011-01-01

    Full Text Available Oral diseases are major health problems with dental caries and periodontal diseases among the most important preventable global infectious diseases. Oral health influences the general quality of life and poor oral health is linked to chronic conditions and systemic diseases. The association between oral diseases and the oral microbiota is well established. Of the more than 750 species of bacteria that inhabit the oral cavity, a number are implicated in oral diseases. The development of dental caries involves acidogenic and aciduric Gram-positive bacteria (mutans streptococci, lactobacilli and actinomycetes. Periodontal diseases have been linked to anaerobic Gram-negative bacteria (Porphyromonas gingivalis, Actinobacillus, Prevotella and Fusobacterium. Given the incidence of oral disease, increased resistance by bacteria to antibiotics, adverse affects of some antibacterial agents currently used in dentistry and financial considerations in developing countries, there is a need for alternative prevention and treatment options that are safe, effective and economical. While several agents are commercially available, these chemicals can alter oral microbiota and have undesirable side-effects such as vomiting, diarrhea and tooth staining. Hence, the search for alternative products continues and natural phytochemicals isolated from plants used as traditional medicines are considered as good alternatives. In this review, plant extracts or phytochemicals that inhibit the growth of oral pathogens, reduce the development of biofilms and dental plaque, influence the adhesion of bacteria to surfaces and reduce the symptoms of oral diseases will be discussed further. Clinical studies that have investigated the safety and efficacy of such plant-derived medicines will also be described.

  13. Broad-Range PCR Coupled with Electrospray Ionization Time of Flight Mass Spectrometry for Detection of Bacteremia and Fungemia in Patients with Neutropenic Fever

    Science.gov (United States)

    Maertens, J.; Bueselinck, K.; Lagrou, K.

    2016-01-01

    Infection is an important complication in patients with hematologic malignancies or solid tumors undergoing intensive cytotoxic chemotherapy. In only 20 to 30% of the febrile neutropenic episodes, an infectious agent is detected by conventional cultures. In this prospective study, the performance of broad-range PCR coupled with electrospray ionization time of flight mass spectrometry (PCR/ESI-MS) technology was compared to conventional blood cultures (BC) in a consecutive series of samples from high-risk hematology patients. In 74 patients, BC and a whole-blood sample for PCR/ESI-MS (Iridica BAC BSI; Abbott, Carlsbad, CA, USA) were collected at the start of each febrile neutropenic episode and, in case of persistent fever, also at day 5. During 100 different febrile episodes, 105 blood samples were collected and analyzed by PCR/ESI-MS. There was evidence of a bloodstream infection (BSI) in 36/105 cases (34%), based on 14 cases with both PCR/ESI-MS and BC positivity, 17 cases with BC positivity only, and 5 cases with PCR/ESI-MS positivity only. The sensitivity of PCR/ESI-MS was 45%, specificity was 93%, and the negative predictive value was 80% compared to blood culture. PCR/ESI-MS detected definite pathogens (Fusobacterium nucleatum and Streptococcus pneumoniae) missed by BC, whereas it missed both Gram-negative and Gram-positive organisms detected by BC. PCR/ESI-MS testing detected additional microorganisms but showed a low sensitivity (45%) compared to BC in neutropenic patients. Our results indicate a lower concordance between BC and PCR/ESI-MS in the neutropenic population than what has been previously reported in other patient groups with normal white blood cell distribution, and a lower sensitivity than other PCR-based methods. PMID:27440820

  14. Analysis of the intestinal lumen microbiota in an animal model of colorectal cancer.

    Directory of Open Access Journals (Sweden)

    Qingchao Zhu

    Full Text Available Recent reports have suggested that multiple factors such as host genetics, environment and diet can promote the progression of healthy mucosa towards sporadic colorectal carcinoma. Accumulating evidence has additionally associated intestinal bacteria with disease initiation and progression. In order to examine and analyze the composition of gut microbiota in the absence of confounding influences, we have established an animal model of 1, 2-dimethylhydrazine (DMH-induced colon cancer. Using this model, we have performed pyrosequencing of the V3 region of the 16S rRNA genes in this study to determine the diversity and breadth of the intestinal microbial species. Our findings indicate that the microbial composition of the intestinal lumen differs significantly between control and tumor groups. The abundance of Firmicutes was elevated whereas the abundance of Bacteroidetes and Spirochetes was reduced in the lumen of CRC rats. Fusobacteria was not detected in any of the healthy rats and there was no significant difference in observed Proteobacteria species when comparing the bacterial communities between our two groups. Interestingly, the abundance of Proteobacteria was higher in CRC rats. At the genus level, Bacteroides exhibited a relatively higher abundance in CRC rats compared to controls (14.92% vs. 9.22%, p<0.001. Meanwhile, Prevotella (55.22% vs. 26.19%, Lactobacillus (3.71% vs. 2.32% and Treponema (3.04% vs. 2.43%, were found to be significantly more abundant in healthy rats than CRC rats (p<0.001, respectively. We also demonstrate a significant reduction of butyrate-producing bacteria such as Roseburia and Eubacterium in the gut microbiota of CRC rats. Furthermore, a significant increase in Desulfovibrio, Erysipelotrichaceae and Fusobacterium was also observed in the tumor group. A decrease in probiotic species such as Ruminococcus and Lactobacillus was likewise observed in the tumor group. Collectively, we can conclude that a significant

  15. Features of the bronchial bacterial microbiome associated with atopy, asthma, and responsiveness to inhaled corticosteroid treatment.

    Science.gov (United States)

    Durack, Juliana; Lynch, Susan V; Nariya, Snehal; Bhakta, Nirav R; Beigelman, Avraham; Castro, Mario; Dyer, Anne-Marie; Israel, Elliot; Kraft, Monica; Martin, Richard J; Mauger, David T; Rosenberg, Sharon R; Sharp-King, Tonya; White, Steven R; Woodruff, Prescott G; Avila, Pedro C; Denlinger, Loren C; Holguin, Fernando; Lazarus, Stephen C; Lugogo, Njira; Moore, Wendy C; Peters, Stephen P; Que, Loretta; Smith, Lewis J; Sorkness, Christine A; Wechsler, Michael E; Wenzel, Sally E; Boushey, Homer A; Huang, Yvonne J

    2017-07-01

    Compositional differences in the bronchial bacterial microbiota have been associated with asthma, but it remains unclear whether the findings are attributable to asthma, to aeroallergen sensitization, or to inhaled corticosteroid treatment. We sought to compare the bronchial bacterial microbiota in adults with steroid-naive atopic asthma, subjects with atopy but no asthma, and nonatopic healthy control subjects and to determine relationships of the bronchial microbiota to phenotypic features of asthma. Bacterial communities in protected bronchial brushings from 42 atopic asthmatic subjects, 21 subjects with atopy but no asthma, and 21 healthy control subjects were profiled by using 16S rRNA gene sequencing. Bacterial composition and community-level functions inferred from sequence profiles were analyzed for between-group differences. Associations with clinical and inflammatory variables were examined, including markers of type 2-related inflammation and change in airway hyperresponsiveness after 6 weeks of fluticasone treatment. The bronchial microbiome differed significantly among the 3 groups. Asthmatic subjects were uniquely enriched in members of the Haemophilus, Neisseria, Fusobacterium, and Porphyromonas species and the Sphingomonodaceae family and depleted in members of the Mogibacteriaceae family and Lactobacillales order. Asthma-associated differences in predicted bacterial functions included involvement of amino acid and short-chain fatty acid metabolism pathways. Subjects with type 2-high asthma harbored significantly lower bronchial bacterial burden. Distinct changes in specific microbiota members were seen after fluticasone treatment. Steroid responsiveness was linked to differences in baseline compositional and functional features of the bacterial microbiome. Even in subjects with mild steroid-naive asthma, differences in the bronchial microbiome are associated with immunologic and clinical features of the disease. The specific differences identified

  16. [Management of Lung Abscess].

    Science.gov (United States)

    Marra, A; Hillejan, L; Ukena, D

    2015-10-01

    A lung abscess is an infectious pulmonary disease characterised by the presence of a pus-filled cavity within the lung parenchyma. The content of an abscess often drains into the airways spontaneously, leading to an air-fluid level visible on chest X-rays and CT scans. Primary lung abscesses occur in patients who are prone to aspiration or in otherwise healthy individuals; secondary lung abscesses typically develop in association with a stenosing lung neoplasm or a systemic disease that compromises immune defences, such as AIDS, or after organ transplantation. The organisms found in abscesses caused by aspiration pneumonia reflect the resident flora of the oropharynx. The most commonly isolated organisms are anaerobic bacteria (Prevotella, Bacteroides, Fusobacterium, Peptostreptococcus) or streptococci; in alcoholics with poor oral hygiene, the spectrum of pathogens includes Staphylococcus aureus, Streptococcus pyogenes and Actinomyces. Chest radiography and computed tomography (CT) are mandatory procedures in the diagnostic algorithm. Standard treatment for a lung abscess consists of systemic antibiotic therapy, which is based on the anticipated or proven bacterial spectrum of the abscess. In most cases, primary abscesses are successfully treated by calculated empiric antibiotic therapy, with an estimated lethality rate of less than 10 %. Secondary abscesses, despite targeted antimicrobial therapy, are associated with a poor prognosis, which depends on the patient's general condition and underlying disease; lethality is as high as 75 %. Negative prognostic factors are old age, severe comorbidities, immunosuppression, bronchial obstruction, and neoplasms. Surgical intervention due to failure of conservative treatment is required in only 10 % of patients, with a success rate of up to 90 % and postoperative mortality rates ranging between 0 and 33 %. Treatment success after endoscopic or percutaneous drainage is achieved in 73 to 100 % of cases, with an

  17. In vitro antioxidant potential of medicinal plant extracts and their activities against oral bacteria based on Brazilian folk medicine.

    Science.gov (United States)

    Alviano, Wagner S; Alviano, Daniela S; Diniz, Cláudio G; Antoniolli, Angelo R; Alviano, Celuta S; Farias, Luiz M; Carvalho, Maria Auxiliadora R; Souza, Margareth M G; Bolognese, Ana Maria

    2008-06-01

    This study aims to determine antibacterial activities of Cocos nucifera (husk fiber), Ziziphus joazeiro (inner bark), Caesalpinia pyramidalis (leaves), aqueous extracts and Aristolochia cymbifera (rhizomes) alcoholic extract against Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium nucleatum, Streptococcus mutans and Lactobacillus casei. The antioxidant activity and acute toxicity of these extracts were also evaluated. The plant extracts antibacterial activity was evaluated in vitro and the minimal inhibitory concentration (MIC) was determined by the broth micro-dilution assay. The bacterial killing kinetic was also evaluated for all extracts. In addition, the antibacterial effect of the extracts was tested in vitro on artificial oral biofilms. The acute toxicity of each extract was determined in according to Lorke [Lorke D. A new approach to practical acute toxicity testing. Arch Toxicol 1983;54:275-87] and the antioxidant activity was evaluated by DPPH photometric assay [Mensor LL, Menezes FS, Leitão GG, Reis AS, Santos TC, Coube CS, et al. Screening of Brazilian plants extract for antioxidant activity by the use of DPPH free radical method. Phytother Res 2001;15:127-30]. MIC and the bactericidal concentrations were identical, for each evaluated extract. However, microbes of artificial biofilms were less sensitive to the extracts than the planktonic strains. A. cymbifera extract induced the highest bactericidal effect against all tested bacteria, followed by C. nucifera, Z. joazeiro and C. pyramidalis extracts, respectively. All extracts showed good antioxidant potential, being C. nucifera and C. pyramidalis aqueous extracts the most active ones. In conclusion, all oral bacteria tested (planktonic or in artificial biofilms) were more susceptible to, and rapidly killed in presence of A. cymbifera, C. pyramidalis and C. nucifera than Z. joazeiro extracts, respectively. Thus, these extracts may be of great interest for future studies about treatment of

  18. Effect of gallium-arsenic laser on photosensitized periodontopathic anaerobic organisms: An in vitro study

    Science.gov (United States)

    Mathur, Setu; Kothiwale, Shaila Veerappa; Nag, Buddhi Prakash; Mathur, Tanu; Bhansali, Ashoka; Khatri, Rohit Kumar

    2016-01-01

    Background: The mainstay of periodontal therapy is mechanical removal of subgingival plaque. There is considerable interest in supplementing it with the use of antibiotics and antiseptics. Many drawbacks are associated with these adjunctive pharmacological regimens such as development of resistance to antibiotics and disruption of microflora of the gastrointestinal tract. Hence, alternate means of killing subgingival bacteria are clearly desirable. One such method is the use of laser. Aim: This study aimed to investigate antibacterial capabilities of gallium-arsenic (Ga-As) laser on photosensitized periodontopathic organisms. The three bacteria selected for the study were Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia. Settings: The subjects for the study were selected from the patients visiting the Department of Periodontics, Karnataka Lingayat Education Society's Institute of Dental Sciences, Belgaum. Design: In vitro study design. Materials and Methods: Subgingival plaque samples collected from chronic periodontitis patients were cultured anaerobically for 72 h. Predetermined number of colonies of each bacterium was taken and was then divided into cases and control groups. Both groups were photosensitized using toluidine blue O (TBO) dye and the case groups were irradiated with Ga-As laser. Bacterial colonies were then serially diluted and were incubated for subculture. After incubation period, the number of viable bacterial count was performed. Statistical Analysis: Wilcoxon-signed rank test was carried out to determine significance of reduction on subsequent dilution within the bacterial group. Mann–Whitney U-test was performed to determine the significance of reduction between cases and control of particular bacterial group. Results: The results revealed substantial reduction in the viable bacterial count. F. nucleatum was found to be most sensitive to killing by laser irradiation followed by P. intermedia and then P

  19. Effect of gallium-arsenic laser on photosensitized periodontopathic anaerobic organisms: An in vitro study

    Directory of Open Access Journals (Sweden)

    Setu Mathur

    2016-01-01

    Full Text Available Background: The mainstay of periodontal therapy is mechanical removal of subgingival plaque. There is considerable interest in supplementing it with the use of antibiotics and antiseptics. Many drawbacks are associated with these adjunctive pharmacological regimens such as development of resistance to antibiotics and disruption of microflora of the gastrointestinal tract. Hence, alternate means of killing subgingival bacteria are clearly desirable. One such method is the use of laser. Aim: This study aimed to investigate antibacterial capabilities of gallium-arsenic (Ga-As laser on photosensitized periodontopathic organisms. The three bacteria selected for the study were Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia. Settings: The subjects for the study were selected from the patients visiting the Department of Periodontics, Karnataka Lingayat Education Society's Institute of Dental Sciences, Belgaum. Design: In vitro study design. Materials and Methods: Subgingival plaque samples collected from chronic periodontitis patients were cultured anaerobically for 72 h. Predetermined number of colonies of each bacterium was taken and was then divided into cases and control groups. Both groups were photosensitized using toluidine blue O (TBO dye and the case groups were irradiated with Ga-As laser. Bacterial colonies were then serially diluted and were incubated for subculture. After incubation period, the number of viable bacterial count was performed. Statistical Analysis: Wilcoxon-signed rank test was carried out to determine significance of reduction on subsequent dilution within the bacterial group. Mann–Whitney U-test was performed to determine the significance of reduction between cases and control of particular bacterial group. Results: The results revealed substantial reduction in the viable bacterial count. F. nucleatum was found to be most sensitive to killing by laser irradiation followed by P. intermedia and

  20. Anaerobic bacteraemia revisited: species and susceptibilities.

    Science.gov (United States)

    Ng, Lily S Y; Kwang, Lee Ling; Rao, Suma; Tan, Thean Yen

    2015-01-01

    This retrospective study was performed to evaluate the frequency of anaerobic bacteraemia over a 10-year period, and to provide updated antibiotic susceptibilities for the more clinically relevant anaerobes causing blood stream infection. Data were retrieved from the laboratory information system for the period 2003 to 2012. During this time, blood cultures were inoculated in Bactec™ Plus vials (BD, USA) and continuously monitored in the Bactec™ 9000 blood culture system (BD, USA). Anaerobic organisms were identified using commercial identification kits, predominantly API 20 A (bioMérieux, France) supplemented with Vitek ANC cards (bioMérieux, France) and AN-Ident discs (Oxoid, United Kingdom). A representative subset of isolates were retrieved from 2009 to 2011 and antimicrobial susceptibilities to penicillin, amoxicillin-clavulanate, clindamycin, imipenem, moxifloxacin, piperacillin-tazobactam and metronidazole were determined using the Etest method. Anaerobes comprised 4.1% of all positive blood culture with 727 obligate anaerobes recovered over the 10-year period, representing a positivity rate of 0.35%. The only significant change in anaerobe positivity rates occurred between 2003 and 2004, with an increase of 0.2%. The Bacteroides fragilis group (45%) were the predominant anaerobic pathogens, followed by Clostridium species (12%), Propioniobacterium species (11%) and Fusobacterium species (6%). The most active in vitro antibiotics were imipenem, piperacillin-tazobactam, amoxicillin-clavulanate and metronidazole, with susceptibilities of 95.0%, 93.3%, 90.8% and 90.8% respectively. Resistance was high to penicillin, clindamycin and moxifl oxacin. However, there were apparent differences for antibiotic susceptibilities between species. This study indicates that the anaerobes comprise a small but constant proportion of bloodstream isolates. Antibiotic resistance was high to some antibiotics, but metronidazole, the beta-lactam/beta-lactamase inhibitors and

  1. Bacterial Community Development in Experimental Gingivitis

    Science.gov (United States)

    Kistler, James O.; Booth, Veronica; Bradshaw, David J.; Wade, William G.

    2013-01-01

    Current knowledge of the microbial composition of dental plaque in early gingivitis is based largely on microscopy and cultural methods, which do not provide a comprehensive description of oral microbial communities. This study used 454-pyrosequencing of the V1–V3 region of 16S rRNA genes (approximately 500 bp), and bacterial culture, to characterize the composition of plaque during the transition from periodontal health to gingivitis. A total of 20 healthy volunteers abstained from oral hygiene for two weeks, allowing plaque to accumulate and gingivitis to develop. Plaque samples were analyzed at baseline, and after one and two weeks. In addition, plaque samples from 20 chronic periodontitis patients were analyzed for cross-sectional comparison to the experimental gingivitis cohort. All of the healthy volunteers developed gingivitis after two weeks. Pyrosequencing yielded a final total of 344 267 sequences after filtering, with a mean length of 354 bases, that were clustered into an average of 299 species-level Operational Taxonomic Units (OTUs) per sample. Principal coordinates analysis (PCoA) plots revealed significant shifts in the bacterial community structure of plaque as gingivitis was induced, and community diversity increased significantly after two weeks. Changes in the relative abundance of OTUs during the transition from health to gingivitis were correlated to bleeding on probing (BoP) scores and resulted in the identification of new health- and gingivitis-associated taxa. Comparison of the healthy volunteers to the periodontitis patients also confirmed the association of a number of putative periodontal pathogens with chronic periodontitis. Taxa associated with gingivitis included Fusobacterium nucleatum subsp. polymorphum, Lachnospiraceae [G-2] sp. HOT100, Lautropia sp. HOTA94, and Prevotella oulorum, whilst Rothia dentocariosa was associated with periodontal health. Further study of these taxa is warranted and may lead to new therapeutic approaches

  2. Bacterial community development in experimental gingivitis.

    Directory of Open Access Journals (Sweden)

    James O Kistler

    Full Text Available Current knowledge of the microbial composition of dental plaque in early gingivitis is based largely on microscopy and cultural methods, which do not provide a comprehensive description of oral microbial communities. This study used 454-pyrosequencing of the V1-V3 region of 16S rRNA genes (approximately 500 bp, and bacterial culture, to characterize the composition of plaque during the transition from periodontal health to gingivitis. A total of 20 healthy volunteers abstained from oral hygiene for two weeks, allowing plaque to accumulate and gingivitis to develop. Plaque samples were analyzed at baseline, and after one and two weeks. In addition, plaque samples from 20 chronic periodontitis patients were analyzed for cross-sectional comparison to the experimental gingivitis cohort. All of the healthy volunteers developed gingivitis after two weeks. Pyrosequencing yielded a final total of 344,267 sequences after filtering, with a mean length of 354 bases, that were clustered into an average of 299 species-level Operational Taxonomic Units (OTUs per sample. Principal coordinates analysis (PCoA plots revealed significant shifts in the bacterial community structure of plaque as gingivitis was induced, and community diversity increased significantly after two weeks. Changes in the relative abundance of OTUs during the transition from health to gingivitis were correlated to bleeding on probing (BoP scores and resulted in the identification of new health- and gingivitis-associated taxa. Comparison of the healthy volunteers to the periodontitis patients also confirmed the association of a number of putative periodontal pathogens with chronic periodontitis. Taxa associated with gingivitis included Fusobacterium nucleatum subsp. polymorphum, Lachnospiraceae [G-2] sp. HOT100, Lautropia sp. HOTA94, and Prevotella oulorum, whilst Rothia dentocariosa was associated with periodontal health. Further study of these taxa is warranted and may lead to new

  3. [Effect of periodontal mechanical treatment on periodontal pathogenic bacteria in gingival crevicular fluid of chronic periodontitis patients].

    Science.gov (United States)

    Ding, Fang; Meng, Huan-xin; Li, Qi-qiang; Zhao, Yi-bing; Feng, Xiang-hui; Zhang, Li

    2010-04-18

    To evaluate the subgingival prevalent rates of 6 periodontal pathogenic bacteria in gingival crevicular fluids of CP patients before and after treatment, to analyze the relationship between the prevalent variance and periodontal clinical parameters, and to provide a microbiologic method of evaluating curative effect and estimating the prognosis. Gingival crevicular fluids of 13 CP patients were collected at baseline, 2 weeks, 2 months and 4 months after periodontal mechanical treatment. Also, gingival crevicular fluids were collected from 11 healthy subjects. Six periodontal pathogenic bacteria including Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis(Pg), Tannerella forsythensis (Tf), Prevotella intermedia (Pi), Fusobacterium nucleatum(Fn), Prevotella nigrescens (Pn) were detected by 16S rRNA based PCR. The PLI, PD, BI of the CP patients 2 months and 4 months after periodontal mechanical treatment were evidently less than those before treatment. These 4 months after treatment were a little more than those 2 months after. The six bacteria were more frequently detected in the CP patients at baseline than in healthy controls. The prevalent rates of Tf (42.1%, 73.7%, 70.2%), Pg (47.4%, 68.4%, 77.2%), Aa (15.8%, 22.8%, 7.0%), Pn (38.6%, 57.9%, 64.9%), Pi(15.8%, 38.6%, 42.1%) 2 weeks, 2 months and 4 months following treatment were significantly lower than those at baseline (Tf 96.5%, Pg 93.0%, Aa 36.8%, Pn 86.0%, Pi 84.2%), but the prevalent rates of all the detected bacteria 2 months after treatment were higher than those at 2 weeks after. Tf, Pg, Aa, Pn and Pi may cooperate in the development of CP. The changes of periodontal pathogenic bacteria could be detected before the changes of clinical parameters and the patients should be re-evaluated and re-treated regularly within 2 months after treatment.

  4. Combined antioxidant effects of Neem extract, bacteria, red blood cells and Lysozyme: possible relation to periodontal disease.

    Science.gov (United States)

    Heyman, Leali; Houri-Haddad, Yael; Heyman, Samuel N; Ginsburg, Isaac; Gleitman, Yossi; Feuerstein, Osnat

    2017-08-10

    The common usage of chewing sticks prepared from Neem tree (Azadirachta indica) in India suggests its potential efficacy in periodontal diseases. The objective of this study is to explore the antibacterial effects of Neem leaf extract on the periodontophatic bacteria Porphyromonas gingivalis and Fusobacterium nucleatum, and its antioxidant capacities alone and in combination with bacteria and polycationic peptides that may be at the site of inflammation. Neem leaf extract was prepared by ethanol extraction. The growth kinetics of P. gingivalis and F. nucleatum under anaerobic conditions in the presence of Neem leaf extract were measured. Broth microdilution test was used to determine the Minimal Inhibitory Concentration (MIC) of Neem leaf extract against each bacterial strain. The effect of Neem leaf extract on the coaggregation of the bacteria was assessed by a visual semi-quantitative assay. The antioxidant capacities of Neem leaf extract alone and in combination with bacteria, with the addition of red blood cells or the polycationic peptides chlorhexidine and lisozyme, were determined using a chemiluminescence assay. Neem leaf extract showed prominent dose-dependent antibacterial activity against P. gingivalis, however, had no effect on the growth of F. nucleatum nor on the coaggregation of the two bacteria. Yet, it showed intense antioxidant activity, which was amplified following adherence to bacteria and with the addition of red blood cells or the polycationic peptides. Neem leaf extract, containing polyphenols that adhere to oral surfaces, have the potential to provide long-lasting antibacterial as well as synergic antioxidant activities when in complex with bacteria, red blood cells and lisozyme. Thus, it might be especially effective in periodontal diseases.

  5. Association of plasma 25-hydroxyvitamin d concentrations and pathogenic oral bacteria in postmenopausal females.

    Science.gov (United States)

    Sahli, Michelle W; Wactawski-Wende, Jean; Ram, Pavani K; LaMonte, Michael J; Hovey, Kathleen M; Genco, Robert J; Andrews, Christopher A; Millen, Amy E

    2014-07-01

    Previous findings of an association between 25-hydroxyvitamin D [25(OH)D] concentrations and periodontal disease may be partially explained by the antimicrobial properties of vitamin D. To the best of the authors' knowledge, no study has investigated the association between 25(OH)D and pathogenic oral bacteria, a putative cause of periodontal disease. The association between plasma 25(OH)D concentrations and pathogenic oral bacteria was examined among postmenopausal females in the Buffalo Osteoporosis and Periodontal Disease Study (1997 to 2000), an ancillary study of the Women's Health Initiative Observational Study. Subgingival plaque samples were assessed using immunofluorescence for the presence of Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Prevotella intermedia, and Campylobacter rectus. Logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for prevalent bacteria by quintile (Q) of 25(OH)D concentrations, adjusting for age and body mass index. Of the 855 participants, 288 (34%) had deficient/inadequate (bacteria. No significant association was found between 25(OH)D and presence of any of these bacteria (adjusted OR for high [Q5] compared to low [Q1] 25(OH)D = 0.96; 95% CI: 0.61 to 1.50; P for trend = 0.50). Inverse, although not statistically significant, associations were found between 25(OH)D and more than one species of pathogenic bacteria (adjusted OR for adequate compared to deficient/inadequate 25(OH)D = 0.85; 95% CI: 0.60 to 1.19). No association was observed between pathogenic oral bacteria and 25(OH)D concentrations in postmenopausal females. This may be attributable to the species of bacteria assessed, small effect size, or a true absence of an association.

  6. Gram-negative periodontal bacteria induce the activation of Toll-like receptors 2 and 4, and cytokine production in human periodontal ligament cells.

    Science.gov (United States)

    Sun, Ying; Shu, Rong; Li, Chao-Lun; Zhang, Ming-Zhu

    2010-10-01

    Periodontitis is a bacterially induced chronic inflammatory disease. Toll-like receptors (TLRs), which could recognize microbial pathogens, are important components in the innate and adaptive immune systems. Both qualitatively and quantitatively distinct immune responses might result from different bacteria stimulation and the triggering of different TLRs. This study explores the interaction of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) with TLR2 and TLR4. We studied the gene expression changes of TLR2 and TLR4 and cytokine production (interleukin-1β, -6, -8, -10, and tumor necrosis factor-alpha) in human periodontal ligament cells (HPDLCs) stimulated with heat-killed bacteria or P. gingivalis lipopolysaccharide (LPS) in the presence or absence of monoclonal antibodies to TLR2 or TLR4 (anti-TLR2/4 mAb). Both test bacteria and 10 microg/ml P. gingivalis LPS treatment increased the gene expression of TLR2 and TLR4 and cytokine production in HPDLCs. In addition, these upregulations could be blocked by anti-TLR2/4 mAb. However, the expression of TLR4 mRNA in HPDLCs stimulated with 1 microg/ml P. gingivalis LPS was not increased. No differences were found in the cytokine production caused by 1 microg/ml P. gingivalis LPS treatment in the presence or absence of anti-TLR4 mAb. These patterns of gene expression and cytokine production indicate that Gram-negative periodontal bacteria or their LPS might play a role in triggering TLR2 and/or TLR4, and be of importance for the immune responses in periodontitis.

  7. Periodontal Pathogens Invade Gingiva and Aortic Adventitia and Elicit Inflammasome Activation in αvβ6 Integrin-Deficient Mice

    Science.gov (United States)

    Velsko, Irina M.; Chukkapalli, Sasanka S.; Rivera-Kweh, Mercedes F.; Zheng, Donghang; Aukhil, Ikramuddin; Lucas, Alexandra R.; Larjava, Hannu

    2015-01-01

    The American Heart Association supports an association between periodontal diseases and atherosclerosis but not a causal association. This study explores the use of the integrin β6−/− mouse model to study the causality. We investigated the ability of a polymicrobial consortium of Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum to colonize the periodontium and induce local and systemic inflammatory responses. Polymicrobially infected Itgβ6−/− mice demonstrate greater susceptibility to gingival colonization/infection, with severe gingival inflammation, apical migration of the junctional epithelium, periodontal pocket formation, alveolar bone resorption, osteoclast activation, bacterial invasion of the gingiva, a greater propensity for the bacteria to disseminate hematogenously, and a strong splenic T cell cytokine response. Levels of atherosclerosis risk factors, including serum nitric oxide, oxidized low-density lipoprotein, serum amyloid A, and lipid peroxidation, were significantly altered by polybacterial infection, demonstrating an enhanced potential for atherosclerotic plaque progression. Aortic gene expression revealed significant alterations in specific Toll-like receptor (TLR) and nucleotide-binding domain- and leucine-rich-repeat-containing receptor (NLR) pathway genes in response to periodontal bacterial infection. Histomorphometry of the aorta demonstrated larger atherosclerotic plaques in Itgβ6−/− mice than in wild-type (WT) mice but no significant difference in atherosclerotic plaque size between mice with polybacterial infection and mice with sham infection. Fluorescence in situ hybridization demonstrated active invasion of the aortic adventitial layer by P. gingivalis. Our observations suggest that polybacterial infection elicits distinct aortic TLR and inflammasome signaling and significantly increases local aortic oxidative stress. These results are the first to demonstrate the

  8. Short-Chain Fatty Acids from Periodontal Pathogens Suppress Histone Deacetylases, EZH2, and SUV39H1 To Promote Kaposi's Sarcoma-Associated Herpesvirus Replication

    Science.gov (United States)

    Yu, Xiaolan; Shahir, Abdel-Malek; Sha, Jingfeng; Feng, Zhimin; Eapen, Betty; Nithianantham, Stanley; Das, Biswajit; Karn, Jonathan; Weinberg, Aaron; Bissada, Nabil F.

    2014-01-01

    ABSTRACT Periodontal pathogens such as Porphyromonas gingivalis and Fusobacterium nucleatum produce five different short-chain fatty acids (SCFAs) as metabolic by-products. We detect significantly higher levels of SCFAs in the saliva of patients with severe periodontal disease. The different SCFAs stimulate lytic gene expression of Kaposi's sarcoma-associated herpesvirus (KSHV) dose dependently and synergistically. SCFAs inhibit class-1/2 histone deacetylases (HDACs) and downregulate expression of silent information regulator-1 (SIRT1). SCFAs also downregulate expression of enhancer of zeste homolog2 (EZH2) and suppressor of variegation 3-9 homolog1 (SUV39H1), which are two histone N-lysine methyltransferases (HLMTs). By suppressing the different components of host epigenetic regulatory machinery, SCFAs increase histone acetylation and decrease repressive histone trimethylations to transactivate the viral chromatin. These new findings provide mechanistic support that SCFAs from periodontal pathogens stimulate KSHV replication and infection in the oral cavity and are potential risk factors for development of oral Kaposi's sarcoma (KS). IMPORTANCE About 20% of KS patients develop KS lesions first in the oral cavity, while other patients never develop oral KS. It is not known if the oral microenvironment plays a role in oral KS tumor development. In this work, we demonstrate that a group of metabolic by-products, namely, short-chain fatty acids, from bacteria that cause periodontal disease promote lytic replication of KSHV, the etiological agent associated with KS. These new findings provide mechanistic support that periodontal pathogens create a unique microenvironment in the oral cavity that contributes to KSHV replication and development of oral KS. PMID:24501407

  9. Regulation of adrenomedullin and nitric oxide production by periodontal bacteria.

    Science.gov (United States)

    Hussain, Q A; McKay, I J; Gonzales-Marin, C; Allaker, R P

    2015-10-01

    In periodontitis the host response to bacterial challenge includes activity of the multifunctional molecules adrenomedullin (AM) and nitric oxide (NO). The aim of this study was to investigate the role of periodontal bacteria in regulating the production of these molecules from cultured cells. Regulation of AM and NO production from oral keratinocytes when challenged with culture supernatants from Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Veillonella atypica, Streptococcus salivarius and Candida albicans was examined. AM and NO were measured in cell culture supernatants using an enzyme-linked immunosorbent assay and the nitrate/nitrite (NO metabolites) Griess assay respectively. Cellular production of AM and inducible NO synthase was also analysed in target cells by immunofluorescence and Western blot analysis. The inter-relationship of AM and NO production were further investigated with macrophages. A. actinomycetemcomitans and C. rectus induced maximal levels of both AM and NO after 6 and 48 h respectively from oral keratinocytes. AM production in macrophages was upregulated in response to the NO donor S-nitrosoglutathione and partially blocked by the inducible NO synthase inhibitor, N(ω) -Nitro-l-arginine methyl ester hydrochloride. Likewise, NO production was increased upon exposure to AM, while the AM receptor antagonist AM 22-52 reduced the release of NO. Pathogens associated with aggressive periodontitis, A. actinomycetemcomitans and C. rectus, were more effective than those associated with chronic periodontitis, P. gingivalis and Prev. intermedia, and commensals, S. salivarius and V. atypica, as regards the upregulation of AM and NO production from oral keratinocytes. Interaction between these molecules was also demonstrated with macrophages. Understanding the coordinated regulation of AM and NO production in response to periodontal bacteria may identify

  10. In vitro evaluation of a multispecies oral biofilm on different implant surfaces

    International Nuclear Information System (INIS)

    Violant, Deborah; Galofré, Marta; Nart, José; Teles, Ricardo Patricio

    2014-01-01

    Biofilm accumulation on implant surfaces is one of the most important factors for early and late implant failure. Because of the related clinical implications, the aim of this in vitro study was to compare the bacterial cell attachment of a four-species oral biofilm on titanium discs of purity grade 2 and 4, with machined surfaces and etched-thermochemically modified with Avantblast®. The in vitro biofilm model was composed of early (Actinomyces naeslundii, Streptococcus gordonii), secondary (Veillonella parvula), and intermediate (Fusobacterium nucleatum ssp. polymorphum) colonizers of tooth surfaces. A total of 36 discs were divided into four groups: Tigr2-c (titanium grade 2, machined surface), Tigr2-t (titanium grade 2, modified surface with Avantblast®), Tigr4-c (titanium grade 4, machined surface), Tigr4-t (titanium grade 4, modified surface with Avantblast®). The experiment was repeated three times. Biofilm viability was tested with 1% 2, 3, 5-triphenyltetrazolium chloride solution and bacterial cell quantification by checkerboard DNA–DNA hybridization. Descriptive analysis was performed to evaluate biofilm composition and differences between groups were checked with the Mann–Whitney test (p < 0.05). After one week, multispecies biofilms showed a similar pattern of bacterial composition on all analyzed implant surfaces. The most prevalent bacterium was V. parvula (∼50% of the total biomass), followed by S. gordonii (∼30%), F. nucleatum ssp. polymorphum (∼10%) and A. naeslundii (<5%). Total bacterial biomass was significantly higher in both grade-4-titanium surfaces (p < 0.05). The results demonstrated that not only implant surface treatment, but also titanium purity, influence early bacterial colonization. (paper)

  11. Extracellular deoxyribonuclease production by periodontal bacteria.

    Science.gov (United States)

    Palmer, L J; Chapple, I L C; Wright, H J; Roberts, A; Cooper, P R

    2012-08-01

    Whilst certain bacteria have long been known to secrete extracellular deoxyribonuclease (DNase), the purpose in microbial physiology was unclear. Recently, however, this enzyme has been demonstrated to confer enhanced virulence, enabling bacteria to evade the host's immune defence of extruded DNA/chromatin filaments, termed neutrophil extracellular traps (NETs). As NETs have recently been identified in infected periodontal tissue, the aim of this study was to screen periodontal bacteria for extracellular DNase activity. To determine whether DNase activity was membrane bound or secreted, 34 periodontal bacteria were cultured in broth and on agar plates. Pelleted bacteria and supernatants from broth cultures were analysed for their ability to degrade DNA, with relative activity levels determined using an agarose gel electrophoresis assay. Following culture on DNA-supplemented agar, expression was determined by the presence of a zone of hydrolysis and DNase activity related to colony size. Twenty-seven bacteria, including red and orange complex members Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Parvimonas micra, Prevotella intermedia, Streptococcus constellatus, Campylobacter rectus and Prevotella nigrescens, were observed to express extracellular DNase activity. Differences in DNase activity were noted, however, when bacteria were assayed in different culture states. Analysis of the activity of secreted DNase from bacterial broth cultures confirmed their ability to degrade NETs. The present study demonstrates, for the first time, that DNase activity is a relatively common property of bacteria associated with advanced periodontal disease. Further work is required to determine the importance of this bacterial DNase activity in the pathogenesis of periodontitis. © 2011 John Wiley & Sons A/S.

  12. Preterm low birthweight and the role of oral bacteria

    Directory of Open Access Journals (Sweden)

    Elizabeth Shira Davenport

    2010-12-01

    Full Text Available Preterm and low birthweight (PTLBW continues to be a major cause of mortality and morbidity across the world. In recent years, maternal periodontal disease has been implicated as a risk factor for adverse pregnancy outcomes. There is conflicting evidence to support such an outcome as illustrated by descriptive, case control and randomised controlled trials involving pregnant women from across the world, using different measurement tools to determine the level of periodontal disease. Whilst considering the literature, there is evidence for both arguments, based on the effect of periodontal inflammatory by products. Bacteria associated with periodontal disease are not dissimilar to those known to be associated with genito-urinary bacterial infections and adverse pregnancy outcomes. Several groups have demonstrated the apparent translocation of Fusobacterium nucleatum, Prevotella nigrescens, Prevotella intermedia, Porphyromonus gingivalis, Treponema denticola to the foetal placental unit whereby a maternal or foetal response has been detected resulting in premature birth or low birthweight. The normal process of parturition involves a cascade of events including a build-up of inflammatory mediators as linked to inflammation, whereby the maternal environment becomes hostile and threatens the well-being of the infant, and the foetus expelled. The question remains therefore, is there a greater risk of delivering a PTLBW infant when the mother has detectable periodontal disease, or is the release of inflammatory mediators and their translocation via the haematogenous route sufficient to induce a poor pregnancy outcome? The data investigated would suggest that there is a positive outcome when certain oral gram-negative bacteria create a cumulative effect sufficient to trigger early delivery, which represents the final straw to result in preterm or low birthweight delivery. There is equally sufficient epidemiological evidence that does not support this

  13. Supernatants from oral epithelial cells and gingival fibroblasts modulate human immunodeficiency virus type 1 promoter activation induced by periodontopathogens in monocytes/macrophages.

    Science.gov (United States)

    González, O A; Ebersole, J L; Huang, C B

    2010-04-01

    Bacterial and host cell products during coinfections of Human Immunodeficiency Virus type 1-positive (HIV-1(+)) patients regulate HIV-1 recrudescence in latently infected cells (e.g. T cells, monocytes/macrophages), impacting highly active antiretroviral therapy (HAART) failure and progression of acquired immunodeficiency syndrome. A high frequency of oral opportunistic infections (e.g. periodontitis) in HIV-1(+) patients has been demonstrated; however, their potential to impact HIV-1 exacerbation is unclear. We sought to determine the ability of supernatants derived from oral epithelial cells (OKF4) and human gingival fibroblasts (Gin-4) challenged with periodontal pathogens, to modulate the HIV-1 promoter activation in monocytes/macrophages. BF24 monocytes/macrophages transfected with the HIV-1 promoter driving the expression of chloramphenicol acetyltransferase (CAT) were stimulated with Porphyromonas gingivalis, Fusobacterium nucleatum, or Treponema denticola in the presence of supernatants from OKF4 or Gin4 cells either unstimulated or previously pulsed with bacteria. CAT levels were determined by enzyme-linked immunosorbent assay and cytokine production was evaluated by Luminex beadlyte assays. OKF4 and Gin4 supernatants enhanced HIV-1 promoter activation particularly related to F. nucleatum challenge. An additive effect was observed in HIV-1 promoter activation when monocytes/macrophages were simultaneously stimulated with gingival cell supernatants and bacterial extracts. OKF4 cells produced higher levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukins -6 and -8 in response to F. nucleatum and P. gingivalis. Preincubation of OKF4 supernatants with anti-GM-CSF reduced the additive effect in periodontopathogen-induced HIV-1 promoter activation. These results suggest that soluble mediators produced by gingival resident cells in response to periodontopathogens could contribute to HIV-1 promoter activation in monocytes

  14. Interleukin-6 production by human monocytes treated with granulocyte-macrophage colony-stimulating factor in the presence of lipopolysaccharide of oral microorganisms.

    Science.gov (United States)

    Baqui, A A; Meiller, T F; Chon, J J; Turng, B F; Falkler, W A

    1998-06-01

    This study focused on the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide of the putative periodontal pathogens Porphyromonas gingivalis or Fusobacterium nucleatum on IL-6 production by THP-1 cells (a human monocytic cell line). Resting THP-1 cells were alternatively treated with GM-CSF (50 IU/ml) and lipopolysaccharide of P. gingivalis or F. nucleatum, in varying concentrations for varying time periods. IL-6 production in supernatant fluids of treated cells was evaluated by an enzyme-linked immunosorbent assay (ELISA) and a reverse transcription polymerase chain reaction (RT-PCR) was used to evaluate gene expression. Untreated THP-1 cells did not produce IL-6 as determined by ELISA. RT-PCR also failed to detect IL-6 mRNA in untreated THP-1 cells, indicating that IL-6 was not constitutively produced. After stimulation of THP-1 cells with lipopolysaccharide of F. nucleatum or P. gingivalis, IL-6 was produced, peaking at 4 h (200-300 pg/ml) and thereafter sharply declining by 8 h. When GM-CSF was added together with lipopolysaccharide of P. gingivalis or F. nucleatum, there was a synergistic quantitative increase in production of IL-6 as measured by ELISA as compared with lipopolysaccharide alone. IL-6 mRNA was detected by RT-PCR, 15 min after stimulation with lipopolysaccharide of either P. gingivalis or F. nucleatum. GM-CSF supplementation with lipopolysaccharide of P. gingivalis shortened the transcription of IL-6 mRNA to 5 min, a shift which was not observed with lipopolysaccharide of F. nucleatum, possibly indicating a different mechanism of initiation of transcription. Production of IL-6 by GM-CSF-treated THP-1 cells in the presence of lipopolysaccharide of oral microorganisms may provide a model for studying the role of macrophages in acute and chronic periodontal diseases, including the clinical periodontal exacerbation as observed in chemotherapy patients receiving GM-CSF for bone marrow recovery.

  15. New bacterial composition in primary and persistent/secondary endodontic infections with respect to clinical and radiographic findings.

    Science.gov (United States)

    Tennert, Christian; Fuhrmann, Maximilian; Wittmer, Annette; Karygianni, Lamprini; Altenburger, Markus J; Pelz, Klaus; Hellwig, Elmar; Al-Ahmad, Ali

    2014-05-01

    The aim of the present study was to analyze the microbiota of primary and secondary/persistent endodontic infections of patients undergoing endodontic treatment with respect to clinical and radiographic findings. Samples from the root canals of 21 German patients were taken using 3 sequential sterile paper points. In the case of a root canal filling, gutta-percha was removed with sterile files, and samples were taken using sterile paper points. The samples were plated, and microorganisms were then isolated and identified morphologically by biochemical analysis and sequencing the 16S rRNA genes of isolated microorganisms. In 12 of 21 root canals, 33 different species could be isolated. Six (50%) of the cases with isolated microorganisms were primary, and 6 (50%) cases were endodontic infections associated with root-filled teeth. Twelve of the isolated species were facultative anaerobic and 21 obligate anaerobic. Monomicrobial infections were found for Enterococcus faecalis and Actinomyces viscosus. E. faecalis was most frequently isolated in secondary endodontic infections (33%). Moraxella osloensis was isolated from a secondary endodontic infection that had an insufficient root canal filling accompanied by a mild sensation of pain. A new bacterial composition compromising Atopobium rimae, Anaerococcus prevotii, Pseudoramibacter alactolyticus, Dialister invisus, and Fusobacterium nucleatum was recovered from teeth with chronic apical abscesses. New bacterial combinations were found and correlated to clinical and radiographic findings, particularly to chronic apical abscesses. M. osloensis was detected in root canals for the second time and only in German patients. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  16. Low antibiotic resistance among anaerobic Gram-negative bacteria in periodontitis 5 years following metronidazole therapy.

    Science.gov (United States)

    Dahlen, G; Preus, H R

    2017-02-01

    The objective of this study was to assess antibiotic susceptibility among predominant Gram-negative anaerobic bacteria isolated from periodontitis patients who 5 years prior had been subject to mechanical therapy with or without adjunctive metronidazole. One pooled sample was taken from the 5 deepest sites of each of 161 patients that completed the 5 year follow-up after therapy. The samples were analyzed by culture. A total number of 85 anaerobic strains were isolated from the predominant subgingival flora of 65/161 patient samples, identified, and tested for antibiotic susceptibility by MIC determination. E-tests against metronidazole, penicillin, amoxicillin, amoxicillin + clavulanic acid and clindamycin were employed. The 73/85 strains were Gram-negative rods (21 Porphyromonas spp., 22 Prevotella/Bacteroides spp., 23 Fusobacterium/Filifactor spp., 3 Campylobacter spp. and 4 Tannerella forsythia). These were all isolated from the treated patients irrespective of therapy procedures (+/-metronidazole) 5 years prior. Three strains (Bifidobacterium spp., Propionibacterium propionicum, Parvimonas micra) showed MIC values for metronidazole over the European Committee on Antimicrobial Susceptibility Testing break point of >4 μg/mL. All Porphyromonas and Tannerella strains were highly susceptible. Metronidazole resistant Gram-negative strains were not found, while a few showed resistance against beta-lactam antibiotics. In this population of 161 patients who had been subject to mechanical periodontal therapy with or without adjunct metronidazole 5 years prior, no cultivable antibiotic resistant anaerobes were found in the predominant subgingival microbiota. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Protective potential of non-dialyzable material fraction of cranberry juice on the virulence of P. gingivalis and F. nucleatum mixed infection.

    Science.gov (United States)

    Polak, David; Naddaf, Raja; Shapira, Lior; Weiss, Ervin I; Houri-Haddad, Yael

    2013-07-01

    Periodontitis is a polymicrobial infectious disease. A novel potential chemical treatment modality may lie in bacterial anti-adhesive materials, such as cranberry juice fractions. The aim of this study is to explore the effect of high molecular weight cranberry constituent (non-dialyzable material [NDM]) on the virulence of a mixed infection with Porphyromonas gingivalis and Fusobacterium nucleatum in mice. In vitro, the anti-adhesive property of NDM was validated on epithelial cell culture, and inhibition of coaggregation was tested using a coaggregation assay. The in vivo effect was tested on the outcome of experimental periodontitis induced by a P. gingivalis and F. nucleatum mixed infection, and also on the local host response using the subcutaneous chamber model of infection. Phagocytosis was also tested on RAW macrophages by the use of fluorescent-labeled bacteria. NDM was found to inhibit the adhesion of both species of bacteria onto epithelial cells and to inhibit coaggregation in a dose-dependent manner. NDM consumption by mice attenuated the severity of experimental periodontitis compared with a mixed infection without NDM treatment. In infected subcutaneous chambers, NDM alone reduced tumor necrosis factor-α (TNF-α) levels induced by the mixed infection. In vitro, NDM eliminated TNF-α expression by macrophages that were exposed to P. gingivalis and F. nucleatum, without impairing their viability. Furthermore, NDM increased the phagocytosis of P. gingivalis. The results indicate that the use of NDM may hold potential protective and/or preventive modalities in periodontal disease. Underlying mechanisms for this trait may perhaps be the anti-adhesive properties of NDM or its potential effect on inflammation.

  18. Prebiotic Potential and Chemical Composition of Seven Culinary Spice Extracts.

    Science.gov (United States)

    Lu, Qing-Yi; Summanen, Paula H; Lee, Ru-Po; Huang, Jianjun; Henning, Susanne M; Heber, David; Finegold, Sydney M; Li, Zhaoping

    2017-08-01

    The objective of this study was to investigate prebiotic potential, chemical composition, and antioxidant capacity of spice extracts. Seven culinary spices including black pepper, cayenne pepper, cinnamon, ginger, Mediterranean oregano, rosemary, and turmeric were extracted with boiling water. Major chemical constituents were characterized by RP-HPLC-DAD method and antioxidant capacity was determined by measuring colorimetrically the extent to scavenge ABTS radical cations. Effects of spice extracts on the viability of 88 anaerobic and facultative isolates from intestinal microbiota were determined by using Brucella agar plates containing serial dilutions of extracts. A total of 14 phenolic compounds, a piperine, cinnamic acid, and cinnamaldehyde were identified and quantitated. Spice extracts exhibited high antioxidant capacity that correlated with the total amount of major chemicals. All spice extracts, with the exception of turmeric, enhanced the growth of Bifidobacterium spp. and Lactobacillus spp. All spices exhibited inhibitory activity against selected Ruminococcus species. Cinnamon, oregano, and rosemary were active against selected Fusobacterium strains and cinnamon, rosemary, and turmeric were active against selected Clostridium spp. Some spices displayed prebiotic-like activity by promoting the growth of beneficial bacteria and suppressing the growth of pathogenic bacteria, suggesting their potential role in the regulation of intestinal microbiota and the enhancement of gastrointestinal health. The identification and quantification of spice-specific phytochemicals provided insight into the potential influence of these chemicals on the gut microbial communities and activities. Future research on the connections between spice-induced changes in gut microbiota and host metabolism and disease preventive effect in animal models and humans is needed. © 2017 The Authors. Journal of Food Science published by Wiley Periodicals, Inc. on behalf of Institute of

  19. Terminal Restriction Fragment Length Polymorphism Analysis of Soil Bacterial Communities under Different Vegetation Types in Subtropical Area.

    Directory of Open Access Journals (Sweden)

    Zeyan Wu

    Full Text Available Soil microbes are active players in energy flow and material exchange of the forest ecosystems, but the research on the relationship between the microbial diversity and the vegetation types is less conducted, especially in the subtropical area of China. In this present study, the rhizosphere soils of evergreen broad-leaf forest (EBF, coniferous forest (CF, subalpine dwarf forest (SDF and alpine meadow (AM were chosen as test sites. Terminal-restriction fragment length polymorphisms (T-RFLP analysis was used to detect the composition and diversity of soil bacterial communities under different vegetation types in the National Natural Reserve of Wuyi Mountains. Our results revealed distinct differences in soil microbial composition under different vegetation types. Total 73 microbes were identified in soil samples of the four vegetation types, and 56, 49, 46 and 36 clones were obtained from the soils of EBF, CF, SDF and AM, respectively, and subsequently sequenced. The Actinobacteria, Fusobacterium, Bacteroidetes and Proteobacteria were the most predominant in all soil samples. The order of Shannon-Wiener index (H of all soil samples was in the order of EBF>CF>SDF>AM, whereas bacterial species richness as estimated by four restriction enzymes indicated no significant difference. Principal component analysis (PCA revealed that the soil bacterial communities' structures of EBF, CF, SDF and AM were clearly separated along the first and second principal components, which explained 62.17% and 31.58% of the total variance, respectively. The soil physical-chemical properties such as total organic carbon (TOC, total nitrogen (TN, total phosphorus (TP and total potassium (TK were positively correlated with the diversity of bacterial communities.

  20. The same microbiota and a potentially discriminant metabolome in the saliva of omnivore, ovo-lacto-vegetarian and Vegan individuals.

    Science.gov (United States)

    De Filippis, Francesca; Vannini, Lucia; La Storia, Antonietta; Laghi, Luca; Piombino, Paola; Stellato, Giuseppina; Serrazanetti, Diana I; Gozzi, Giorgia; Turroni, Silvia; Ferrocino, Ilario; Lazzi, Camilla; Di Cagno, Raffaella; Gobbetti, Marco; Ercolini, Danilo

    2014-01-01

    The salivary microbiota has been linked to both oral and non-oral diseases. Scant knowledge is available on the effect of environmental factors such as long-term dietary choices on the salivary microbiota and metabolome. This study analyzed the microbial diversity and metabolomic profiles of the saliva of 161 healthy individuals who followed an omnivore or ovo-lacto-vegetarian or vegan diet. A large core microbiota was identified, including 12 bacterial genera, found in >98% of the individuals. The subjects could be stratified into three "salivary types" that differed on the basis of the relative abundance of the core genera Prevotella, Streptococcus/Gemella and Fusobacterium/Neisseria. Statistical analysis indicated no effect of dietary habit on the salivary microbiota. Phylogenetic beta-diversity analysis consistently showed no differences between omnivore, ovo-lacto-vegetarian and vegan individuals. Metabolomic profiling of saliva using (1)H-NMR and GC-MS/SPME identified diet-related biomarkers that enabled a significant discrimination between the 3 groups of individuals on the basis of their diet. Formate, urea, uridine and 5-methyl-3-hexanone could discriminate samples from omnivores, whereas 1-propanol, hexanoic acid and proline were characteristic of non-omnivore diets. Although the salivary metabolome can be discriminating for diet, the microbiota has a remarkable inter-individual stability and did not vary with dietary habits. Microbial homeostasis might be perturbed with sub-standard oral hygiene or other environmental factors, but there is no current indication that a choice of an omnivore, ovo-lacto-vegetarian or vegan diet can lead to a specific composition of the oral microbiota with consequences on the oral homeostasis.

  1. Extensive Identification of Bacterial Riboflavin Transporters and Their Distribution across Bacterial Species

    Science.gov (United States)

    Merino, Enrique; Bonomi, Hernán Ruy; Goldbaum, Fernando Alberto; García-Angulo, Víctor Antonio

    2015-01-01

    Riboflavin, the precursor for the cofactors flavin mononucleotide (FMN) and flavin adenine dinucleotide, is an essential metabolite in all organisms. While the functions for de novo riboflavin biosynthesis and riboflavin import may coexist in bacteria, the extent of this co-occurrence is undetermined. The RibM, RibN, RfuABCD and the energy-coupling factor-RibU bacterial riboflavin transporters have been experimentally characterized. In addition, ImpX, RfnT and RibXY are proposed as riboflavin transporters based on positional clustering with riboflavin biosynthetic pathway (RBP) genes or conservation of the FMN riboswitch regulatory element. Here, we searched for the FMN riboswitch in bacterial genomes to identify genes encoding riboflavin transporters and assessed their distribution among bacteria. Two new putative riboflavin transporters were identified: RibZ in Clostridium and RibV in Mesoplasma florum. Trans-complementation of an Escherichia coli riboflavin auxotroph strain confirmed the riboflavin transport activity of RibZ from Clostridium difficile, RibXY from Chloroflexus aurantiacus, ImpX from Fusobacterium nucleatum and RfnT from Ochrobactrum anthropi. The analysis of the genomic distribution of all known bacterial riboflavin transporters revealed that most occur in species possessing the RBP and that some bacteria may even encode functional riboflavin transporters from two different families. Our results indicate that some species possess ancestral riboflavin transporters, while others possess transporters that appear to have evolved recently. Moreover, our data suggest that unidentified riboflavin transporters also exist. The present study doubles the number of experimentally characterized riboflavin transporters and suggests a specific, non-accessory role for these proteins in riboflavin-prototrophic bacteria. PMID:25938806

  2. Microbial changes in conjunctival flora with 30-day continuous-wear silicone hydrogel contact lenses.

    Science.gov (United States)

    Iskeleli, Güzin; Bahar, Hrisi; Eroglu, Ebru; Torun, Muzeyyen Mamal; Ozkan, Sehirbay

    2005-05-01

    To determine the effect of 30-day continuous-wear silicone hydrogel contact lenses on the conjunctival flora in asymptomatic wearers. The authors studied 29 eyes of 15 patients wearing Focus NIGHT & DAY silicone hydrogel contact lenses for up to 30 nights of continuous wear. The average age of the patients was 25.54 +/- 8.98 years. Cultures of the inferior cul-de-sac were taken bilaterally from all eyes, before and after lens wear in asymptomatic patients. The isolation and identification of bacteria were made by standard clinical laboratory methods. The number of eyes whose conjunctival cultures were sterile before using the lenses significantly decreased (P = 0.0005), and the number of eyes with a growth of coagulase-negative staphylococci and diphtheroid rods in their conjunctival cultures significantly increased after using these lenses (P = 0.001 and P = 0.031, respectively). Conversely, a statistically significant difference was not found in the number of eyes that carried Propionibacterium acnes and Fusobacterium nucleatum in their conjunctival cultures before and after using the 30-day continuous-wear silicone hydrogel lenses (P = 0.998 and P = 0.488, respectively). The results suggest that the sterility of the conjunctiva significantly decreased after using 30-day continuous-wear silicone hydrogel contact lenses. In addition, the number of bacteria of the normal conjunctival flora significantly increased after the use of these lenses. Contamination by the bacteria of the eyelids may be a possible colonization factor in this study group. Therefore, it is appropriate to examine the patients who wear these lenses more frequently.

  3. Novel Model for Multispecies Biofilms That Uses Rigid Gas-Permeable Lenses ▿

    Science.gov (United States)

    Peyyala, Rebecca; Kirakodu, Sreenatha S.; Ebersole, Jeffrey L.; Novak, Karen F.

    2011-01-01

    Oral biofilms comprise complex multispecies consortia aided by specific inter- and intraspecies interactions occurring among commensals and pathogenic bacterial species. Oral biofilms are primary initiating factors of periodontal disease, although complex multifactorial biological influences, including host cell responses, contribute to the individual outcome of the disease. To provide a system to study initial stages of interaction between oral biofilms and the host cells that contribute to the disease process, we developed a novel in vitro model system to grow biofilms on rigid gas-permeable contact lenses (RGPLs), which enable oxygen to permeate through the lens material. Bacterial species belonging to early- and late-colonizing groups were successfully established as single- or three-species biofilms, with each group comprising Streptococcus gordonii, Streptococcus oralis, and Streptococcus sanguinis; S. gordonii, Actinomyces naeslundii, and Fusobacterium nucleatum; or S. gordonii, F. nucleatum, and Porphyromonas gingivalis. Quantification of biofilm numbers by quantitative PCR (qPCR) revealed substantial differences in the magnitude of bacterial numbers in single-species and multispecies biofilms. We evaluated cell-permeable conventional nucleic acid stains acridine orange, hexidium iodide, and Hoechst 33258 and novel SYTO red, blue, and green fluorochromes for their effect on bacterial viability and fluorescence yield to allow visualization of the aggregates of individual bacterial species by confocal laser scanning microscopy (CLSM). Substantial differences in the quantity and distribution of the species in the multispecies biofilms were identified. The specific features of these biofilms may help us better understand the role of various bacteria in local challenge of oral tissues. PMID:21421785

  4. Periowave demonstrates bactericidal activity against periopathogens and leads to improved clinical outcomes in the treatment of adult periodontitis

    Science.gov (United States)

    Street, Cale N.; Andersen, Roger; Loebel, Nicolas G.

    2009-02-01

    Periodontitis affects half of the U.S. population over 50, and is the leading cause of tooth loss after 35. It is believed to be caused by growth of complex bacterial biofilms on the tooth surface below the gumline. Photodynamic therapy, a technology used commonly in antitumor applications, has more recently been shown to exhibit antimicrobial efficacy. We have demonstrated eradication of the periopathogens Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans in vitro using PeriowaveTM; a commercial photodisinfection system. In addition, several clinical studies have now demonstrated the efficacy of this treatment. A pilot study in the U.S. showed that 68% of patients treated with PeriowaveTM adjunctively to scaling and root planing (SRP) showed clinical attachment level increase of >1 mm, as opposed to 30% with SRP alone. In a subsequent larger study, a second PeriowaveTM treatment 6 weeks after initial treatment led to pocket depth improvements of >1.5 mm in 89% of patients. Finally, in the most recent multicenter, randomized, examiner-blinded study conducted on 121 subjects in Canada, PeriowaveTM treatment produced highly significant gains in attachment level (0.88 mm vs. 0.57 mm; p=0.003) and pocket depth (0.87 mm vs. 0.63 mm; p=0.01) as compared to SRP alone. In summary, PeriowaveTM demonstrated strong bactericidal activity against known periopathogens, and treatment of periodontitis using this system produced significantly better clinical outcomes than SRP alone. This, along with the absence of any adverse events in patients treated to date demonstrates that PDT is a safe and effective treatment for adult chronic periodontitis.

  5. Periodontitis induced by Porphyromonas gingivalis drives periodontal microbiota dysbiosis and insulin resistance via an impaired adaptive immune response.

    Science.gov (United States)

    Blasco-Baque, Vincent; Garidou, Lucile; Pomié, Céline; Escoula, Quentin; Loubieres, Pascale; Le Gall-David, Sandrine; Lemaitre, Mathieu; Nicolas, Simon; Klopp, Pascale; Waget, Aurélie; Azalbert, Vincent; Colom, André; Bonnaure-Mallet, Martine; Kemoun, Philippe; Serino, Matteo; Burcelin, Rémy

    2017-05-01

    To identify a causal mechanism responsible for the enhancement of insulin resistance and hyperglycaemia following periodontitis in mice fed a fat-enriched diet. We set-up a unique animal model of periodontitis in C57Bl/6 female mice by infecting the periodontal tissue with specific and alive pathogens like Porphyromonas gingivalis ( Pg ), Fusobacterium nucleatum and Prevotella intermedia . The mice were then fed with a diabetogenic/non-obesogenic fat-enriched diet for up to 3 months. Alveolar bone loss, periodontal microbiota dysbiosis and features of glucose metabolism were quantified. Eventually, adoptive transfer of cervical (regional) and systemic immune cells was performed to demonstrate the causal role of the cervical immune system. Periodontitis induced a periodontal microbiota dysbiosis without mainly affecting gut microbiota. The disease concomitantly impacted on the regional and systemic immune response impairing glucose metabolism. The transfer of cervical lymph-node cells from infected mice to naive recipients guarded against periodontitis-aggravated metabolic disease. A treatment with inactivated Pg prior to the periodontal infection induced specific antibodies against Pg and protected the mouse from periodontitis-induced dysmetabolism. Finally, a 1-month subcutaneous chronic infusion of low rates of lipopolysaccharides from Pg mimicked the impact of periodontitis on immune and metabolic parameters. We identified that insulin resistance in the high-fat fed mouse is enhanced by pathogen-induced periodontitis. This is caused by an adaptive immune response specifically directed against pathogens and associated with a periodontal dysbiosis. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  6. Effects by periodontitis on pristane-induced arthritis in rats.

    Science.gov (United States)

    Eriksson, Kaja; Lönnblom, Erik; Tour, Gregory; Kats, Anna; Mydel, Piotr; Georgsson, Pierre; Hultgren, Catharina; Kharlamova, Nastya; Norin, Ulrika; Jönsson, Jörgen; Lundmark, Anna; Hellvard, Annelie; Lundberg, Karin; Jansson, Leif; Holmdahl, Rikard; Yucel-Lindberg, Tülay

    2016-11-03

    An infection-immune association of periodontal disease with rheumatoid arthritis has been suggested. This study aimed to investigate the effect of pre-existing periodontitis on the development and the immune/inflammatory response of pristane-induced arthritis. We investigated the effect of periodontitis induced by ligature placement and Porphyromonas gingivalis (P. gingivalis) infection, in combination with Fusobacterium nucleatum to promote its colonization, on the development of pristane-induced arthritis (PIA) in rats (Dark Agouti). Disease progression and severity of periodontitis and arthritis was monitored using clinical assessment, micro-computed tomography (micro-CT)/intraoral radiographs, antibody response, the inflammatory markers such as α-1-acid glycoprotein (α-1-AGP) and c-reactive protein (CRP) as well as cytokine multiplex profiling at different time intervals after induction. Experimentally induced periodontitis manifested clinically (P periodontitis-induction led to severe arthritis in all rats demonstrating that the severity of arthritis was not affected by the pre-existence of periodontitis. Endpoint analysis showed that 89% of the periodontitis-affected animals were positive for antibodies against arginine gingipain B and furthermore, the plasma antibody levels to a citrullinated P. gingivalis peptidylarginine deiminase (PPAD) peptide (denoted CPP3) were significantly (P periodontitis rats with PIA. Additionally, there was a trend towards increased pro-inflammatory and anti-inflammatory cytokine levels, and increased α-1-AGP levels in plasma from periodontitis-challenged PIA rats. Pre-existence of periodontitis induced antibodies against citrullinated peptide derived from PPAD in rats with PIA. However, there were no differences in the development or severity of PIA between periodontitis challenged and periodontitis free rats.

  7. An investigation of the presence of specific anaerobic species in necrotic primary teeth

    Directory of Open Access Journals (Sweden)

    Genara Brum Gomes

    2013-04-01

    Full Text Available Different microbial identification methods have shown that the microbial community profiles in endodontic infections are diverse and assorted. The aim of this study was to evaluate the frequency of selected endodontic pathogens in the pulp chambers (PCs and root canals (RCs of infected primary teeth using PCR methods. Paired PC and RC samples were collected from 15 subjects and analyzed by PCR for the presence of Filifactor alocis, Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella tannerae, Tanerella forsythia, Treponema denticola, and Treponema socranskii. The frequency of each species was determined in the PC and RC of each case. The species most frequently detected in PCs were P. nigrescens (86.7%, P. gingivalis (73.3%, and F. alocis (73.3%. Of the PC samples, 13.3% contained P. micra and T. denticola, and 6.7% contained T. forsythia. The species most frequently detected in RCs were P. gingivalis (100% and P. nigrescens (93.3%. P. tannerae, P. micra, and T. denticola were found in 40% of the RC samples; T. forsythia was found in 26.7% of the RC samples. The “red complex”, which comprises P. gingivalis, T. denticola, and T. forsythia, was not found in the PC of any tooth but was found in 30% of the RC samples. The detection of P. nigrescens in the PC was statistically associated with the presence of P. nigrescens in the RC (p = 0.04. The results suggest high heterogeneity among the samples, even among those from the same subject.

  8. Synergistic Antibacterial Effects of Nanoparticles Encapsulated with Scutellaria baicalensis and Pure Chlorhexidine on Oral Bacterial Biofilms

    Directory of Open Access Journals (Sweden)

    Ken Cham-Fai Leung

    2016-04-01

    Full Text Available Scutellaria baicalensis (SB is a traditional Chinese medicine for treating infectious and inflammatory diseases. Our recent study shows potent antibacterial effects of nanoparticle-encapsulated chlorhexidine (Nano-CHX. Herein, we explored the synergistic effects of the nanoparticle-encapsulated SB (Nano-SB and Nano-CHX on oral bacterial biofilms. Loading efficiency of Nano-SB was determined by thermogravimetric analysis, and its releasing profile was assessed by high-performance liquid chromatographyusing baicalin (a flavonoid compound of SB as the marker. The mucosal diffusion assay on Nano-SB was undertaken in a porcine model. The antibacterial effects of the mixed nanoparticles (Nano-MIX of Nano-SB and Nano-CHX at 9:1 (w/w ratio were analyzed in both planktonic and biofilm modes of representative oral bacteria. The Nano-MIX was effective on the mono-species biofilms of Streptococcus (S. mutans, S. sobrinus, Fusobacterium (F. nucleatum, and Aggregatibacter (A. actinomycetemcomitans (MIC 50 μg/mL at 24 h, and exhibited an enhanced effect against the multi-species biofilms such as S. mutans, F. nucleatum, A. actinomycetemcomitans, and Porphyromonas (P. gingivalis (MIC 12.5 μg/mL at 24 h that was supported by the findings of both scanning electron microscopy (SEM and confocal scanning laser microscopy (CLSM. This study shows enhanced synergistic antibacterial effects of the Nano-MIX on common oral bacterial biofilms, which could be potentially developed as a novel antimicrobial agent for clinical oral/periodontal care.

  9. Impact of early colonizers on in vitro subgingival biofilm formation.

    Directory of Open Access Journals (Sweden)

    Thomas W Ammann

    Full Text Available The aim of this study was to investigate the impact of early colonizing species on the structure and the composition of the bacterial community developing in a subgingival 10-species biofilm model system. The model included Streptococcus oralis, Streptococcus anginosus, Actinomycesoris, Fusobacterium nucleatum subsp. nucleatum, Veillonella dispar, Campylobacter rectus, Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. Based on literature, we considered Streptococcus oralis, Streptococcus anginosus, and Actinomyces oris as early colonizers and examined their role in the biofilms by either a delayed addition to the consortium, or by not inoculating at all the biofilms with these species. We quantitatively evaluated the resulting biofilms by real-time quantitative PCR and further compared the structures using confocal laser scanning microscopy following fluorescence in situ hybridisation. The absence of the early colonizers did not hinder biofilm formation. The biofilms reached the same total counts and developed to normal thickness. However, quantitative shifts in the abundances of individual species were observed. In the absence of streptococci, the overall biofilm structure appeared looser and more dispersed. Moreover, besides a significant increase of P. intermedia and a decrease of P. gingivalis , P. intermedia appeared to form filamented long chains that resembled streptococci. A. oris, although growing to significantly higher abundance in absence of streptococci, did not have a visible impact on the biofilms. Hence, in the absence of the early colonizers, there is a pronounced effect on P. intermedia and P. gingivalis that may cause distinct shifts in the structure of the biofilm. Streptococci possibly facilitate the establishment of P. gingivalis into subgingival biofilms, while in their absence P. intermedia became more dominant and forms elongated chains.

  10. Effect of controlled-release PeriochipTM on clinical and microbiological parameters in patients of chronic periodontitis

    Directory of Open Access Journals (Sweden)

    Komal Puri

    2013-01-01

    Full Text Available Background: The aim of the present study was to evaluate and compare the clinical and microbiological effectiveness of Periochip TM as an adjunct to scaling and root planing (SRP with SRP alone in patients with chronic periodontitis. Materials and Methods: This randomized, split mouth, 3-month clinical and microbiological trial included 30 sites in 15 patients aged 30-50 years diagnosed with chronic periodontitis. In each patient, two bilateral pockets probing 5-7 mm were randomly assigned to test and control groups. The test group received SRP plus Periochip TM , whereas the control group received SRP alone. Clinical indices and anaerobic culture analysis was done at baseline, 1 month, and 3 months interval. Total bacterial count and analysis of four major periodontopathogenic bacteria Porphyromonas gingivalis (Pg, Prevotella intermedia (Pi, Aggregatibacter actinomycetemcomitans (Aa, and Fusobacterium nucleatum (Fn was done. Results: Significant improvement was obtained in all clinical variables in the test group as compared to the control group over the study period. Total colony counts were significantly reduced in the test group as compared to control over the period of time. At baseline Aa was recovered from 4 test group sites and 5 control group sites, Pg from 15 test group and 14 control group sites, Pi from 5 test group and 2 control group sites, Fn from 7 test and 7 control group sites. At 3 months, Aa was recovered from 1 test group and 4 control group sites, Pg from 4 test group and 8 control group sites, Pi from 1 test group and 1 control group site, Fn from 3 test and 4 control group sites. Conclusion: Periochip TM placement as an adjunct to SRP, showed promising results, when compared to SRP alone. Healthy microflora can be maintained for a longer period of time and delay in the repopulation by periodontopathic microorganisms was observed.

  11. The oral microbiome and adverse pregnancy outcomes

    Directory of Open Access Journals (Sweden)

    Cobb CM

    2017-08-01

    Full Text Available Charles M Cobb,1 Patricia J Kelly,2 Karen B Williams,3 Shilpa Babbar,4 Mubashir Angolkar,5 Richard J Derman6 1Department of Periodontics, School of Dentistry, 2Department of Public Health Nursing, School of Nursing and Health Studies, 3Department of Biomedical & Health Informatics, School of Medicine, University of Missouri-Kansas City, Kansas City, MO, 4Department of Obstetrics, Gynecology & Women’s Health, Division of Maternal & Fetal Medicine, School of Medicine, Saint Louis University, St Louis, MO, USA; 5Department of Public Health, Jawaharlal Nehru Medical College (JNMC, KLE University, Karnataka, India; 6Department of Obstetrics & Gynecology, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA, USA Abstract: Significant evidence supports an association between periodontal pathogenic bacteria and preterm birth and preeclampsia. The virulence properties assigned to specific oral pathogenic bacteria, for example, Fusobacterium nucleatum, Porphyromonas gingivalis, Filifactor alocis, Campylobacter rectus, and others, render them as potential collaborators in adverse outcomes of pregnancy. Several pathways have been suggested for this association: 1 hematogenous spread (bacteremia of periodontal pathogens; 2 hematogenous spread of multiple mediators of inflammation that are generated by the host and/or fetal immune response to pathogenic bacteria; and 3 the possibility of oral microbial pathogen transmission, with subsequent colonization, in the vaginal microbiome resulting from sexual practices. As periodontal disease is, for the most part, preventable, the medical and dental public health communities can address intervention strategies to control oral inflammatory disease, lessen the systemic inflammatory burden, and ultimately reduce the potential for adverse pregnancy outcomes. This article reviews the oral, vaginal, and placental microbiomes, considers their potential impact on preterm labor, and the future

  12. Mechanisms by which Porphyromonas gingivalis evades innate immunity.

    Directory of Open Access Journals (Sweden)

    Kaveh Abdi

    Full Text Available The oral cavity is home to unique resident microbial communities whose interactions with host immunity are less frequently studied than those of the intestinal microbiome. We examined the stimulatory capacity and the interactions of two oral bacteria, Porphyromonas gingivalis (P. gingivalis and Fusobacterium nucleatum (F. nucleatum, on Dendritic Cell (DC activation, comparing them to the effects of the well-studied intestinal microbe Escherichia coli (E. coli. Unlike F. nucleatum and E. coli, P. gingivalis failed to activate DCs, and in fact silenced DC responses induced by F. nucleatum or E. coli. We identified a variant strain of P. gingivalis (W50 that lacked this immunomodulatory activity. Using biochemical approaches and whole genome sequencing to compare the two substrains, we found a point mutation in the hagA gene. This protein is though to be involved in the alteration of the PorSS/gingipain pathway, which regulates protein secretion into the extracellular environment. A proteomic comparison of the secreted products of the two substrains revealed enzymatic differences corresponding to this phenotype. We found that P. gingivalis secretes gingipain(s that inactivate several key proinflammatory mediators made by DCs and/or T cells, but spare Interleukin-1 (IL-1 and GM-CSF, which can cause capillary leaks that serve as a source of the heme that P. gingivalis requires for its survival, and GM-CSF, which can cause epithelial-cell growth. Taken together, our results suggest that P. gingivalis has evolved potent mechanisms to modulate its virulence factors and dampen the innate immune response by selectively inactivating most proinflammatory cytokines.

  13. A new type of Na(+-driven ATP synthase membrane rotor with a two-carboxylate ion-coupling motif.

    Directory of Open Access Journals (Sweden)

    Sarah Schulz

    Full Text Available The anaerobic bacterium Fusobacterium nucleatum uses glutamate decarboxylation to generate a transmembrane gradient of Na⁺. Here, we demonstrate that this ion-motive force is directly coupled to ATP synthesis, via an F₁F₀-ATP synthase with a novel Na⁺ recognition motif, shared by other human pathogens. Molecular modeling and free-energy simulations of the rotary element of the enzyme, the c-ring, indicate Na⁺ specificity in physiological settings. Consistently, activity measurements showed Na⁺ stimulation of the enzyme, either membrane-embedded or isolated, and ATP synthesis was sensitive to the Na⁺ ionophore monensin. Furthermore, Na⁺ has a protective effect against inhibitors targeting the ion-binding sites, both in the complete ATP synthase and the isolated c-ring. Definitive evidence of Na⁺ coupling is provided by two identical crystal structures of the c₁₁ ring, solved by X-ray crystallography at 2.2 and 2.6 Å resolution, at pH 5.3 and 8.7, respectively. Na⁺ ions occupy all binding sites, each coordinated by four amino acids and a water molecule. Intriguingly, two carboxylates instead of one mediate ion binding. Simulations and experiments demonstrate that this motif implies that a proton is concurrently bound to all sites, although Na⁺ alone drives the rotary mechanism. The structure thus reveals a new mode of ion coupling in ATP synthases and provides a basis for drug-design efforts against this opportunistic pathogen.

  14. Microbiota epitope similarity either dampens or enhances the immunogenicity of disease-associated antigenic epitopes.

    Directory of Open Access Journals (Sweden)

    Sebastian Carrasco Pro

    Full Text Available The microbiome influences adaptive immunity and molecular mimicry influences T cell reactivity. Here, we evaluated whether the sequence similarity of various antigens to the microbiota dampens or increases immunogenicity of T cell epitopes. Sets of epitopes and control sequences derived from 38 antigenic categories (infectious pathogens, allergens, autoantigens were retrieved from the Immune Epitope Database (IEDB. Their similarity to microbiome sequences was calculated using the BLOSUM62 matrix. We found that sequence similarity was associated with either dampened (tolerogenic; e.g. most allergens or increased (inflammatory; e.g. Dengue and West Nile viruses likelihood of a peptide being immunogenic as a function of epitope source category. Ten-fold cross-validation and validation using sets of manually curated epitopes and non-epitopes derived from allergens were used to confirm these initial observations. Furthermore, the genus from which the microbiome homologous sequences were derived influenced whether a tolerogenic versus inflammatory modulatory effect was observed, with Fusobacterium most associated with inflammatory influences and Bacteroides most associated with tolerogenic influences. We validated these effects using PBMCs stimulated with various sets of microbiome peptides. "Tolerogenic" microbiome peptides elicited IL-10 production, "inflammatory" peptides elicited mixed IL-10/IFNγ production, while microbiome epitopes homologous to self were completely unreactive for both cytokines. We also tested the sequence similarity of cockroach epitopes to specific microbiome sequences derived from households of cockroach allergic individuals and non-allergic controls. Microbiomes from cockroach allergic households were less likely to contain sequences homologous to previously defined cockroach allergens. These results are compatible with the hypothesis that microbiome sequences may contribute to the tolerization of T cells for allergen

  15. A STUDY OF PREVALENCE OF BACTERIAL VAGINOSIS IN SEXUALLY ACTIVE FEMALES- A CROSS-SECTIONAL STUDY IN TERTIARY CARE HOSPITAL, GAYA

    Directory of Open Access Journals (Sweden)

    Supriya Krishna

    2018-01-01

    Full Text Available BACKGROUND Vaginal discharge is very common problem among females. Alteration in balance of normal vaginal organisms can cause the overgrowth of the bacteria that creates vaginal discharge. It is common among sexually active women yet there still remain gaps in our knowledge of this infectious disorder. Bacterial Vaginosis (BV also called no-specific vaginitis develops when the normally predominant peroxide producing lactobacillus species in the vagina are replaced by mixed predominantly anaerobic flora consisting of Gardnerella vaginalis, Mycoplasma hominis, Mobiluncus species, Bacteroides species, Prevotela species, Peptostreptococcus species, Fusobacterium species and Porphyromonas species. The present study was conducted to know the prevalence of bacterial vaginosis in sexually active females coming with the chief complaint of vaginal discharge to Obstetrics and Gynaecological OPD Bihar. MATERIALS AND METHODS A total of 200 otherwise healthy women of reproductive age group with the complaint of excessive vaginal discharge were included in the study. We exclude the patients who are married, women with known skin disease and post-menopausal. Normal saline wet-mount slide preparations were made for detection of motile trichomonads. Gram stained smear were prepared and scored as per classification developed by Nugent. The presence of pseudohyphae and/or budding yeast cells was considered diagnostic of candidal infection. RESULTS The median age of the study population was 28 years. Most common cause was bacterial vaginosis (positive= 18.5 %; intermediate score= 19.5 %, followed by candidiasis (13.5% and trichomoniasis (0.5%. No etiological diagnosis for vaginal discharge could be established in approximately half of the women. Only one woman was HIV positive; one was reactive by VDRL and TPHA tests. CONCLUSION In addition, the laboratory services network needs to be strengthened to ensure accurate and standardized availability of diagnostic

  16. Effects of Repeated Screw Tightening on Implant Abutment Interfaces in Terms of Bacterial and Yeast Leakage in Vitro: One-Time Abutment Versus the Multiscrewing Technique.

    Science.gov (United States)

    Calcaterra, Roberta; Di Girolamo, Michele; Mirisola, Concetta; Baggi, Luigi

    2016-01-01

    Screw loosening can damage the interfaces of implant components, resulting in susceptibility to contamination of the internal parts by microorganisms. The aim of this study was to investigate the impact of abutment screw retightening on the leakage of two different types of bacteria, Streptococcus sanguinis and Fusobacterium nucleatum, and of the yeast Candida albicans. Two types of implant-abutment systems with tube-in-tube interfaces were tested. Groups A and B each used a different type of system that consisted of 20 different pieces that were assembled according to the manufacturer's torque recommendations; four samples in each group were closed just one time, four samples three times, four samples five times, four samples seven times, and four samples nine times. The implants of groups A and B were contaminated with 0.1 μL of microbial solution just before being assembled for the last time to minimize the possibility of contamination. Results showed a direct correlation between the number of colony-forming units grown in the plates and the closing/opening cycles of the implant-abutment systems. Within the limitations of this study, the results indicate the possibility that repeated closing/opening cycles of the implant-abutment unit may influence bacterial/yeast leakage, most likely as a consequence of decreased precision of the coupling between the abutment and the internal part of the dental implant. These findings suggest that a one-time abutment technique may avoid microbiologic leakage in cases of implant-abutment systems with tube-in-tube interfaces.

  17. Microbiological effects and recolonization patterns after adjunctive subgingival debridement with Er:YAG laser.

    Science.gov (United States)

    Sanz-Sánchez, Ignacio; Ortiz-Vigón, Alberto; Herrera, David; Sanz, Mariano

    2016-07-01

    The objective of this study was to assess the microbiological effects and recolonization patterns after non-surgical periodontal treatment protocol based on the adjunctive use of erbium-doped yttrium aluminium garnet (Er:YAG) laser. Patients diagnosed with chronic periodontitis were randomly assigned to two different treatment protocols: test, full-mouth subgingival ultrasonic instrumentation followed by Er-YAG laser application 1 week later to sites with initial probing pocket depth ≥4.5 mm; and control, full-mouth ultrasonic subgingival instrumentation within 1 week. Clinical (at sampled sites) and microbiological (culture-based) parameters were recorded at baseline and 3 and 12 months. Microbiological variables included total counts, frequency of detection, proportions and counts of target species. Results from 19 test and 21 control patients were compared. Minor changes were observed for total colony-forming units, with no differences between groups. For the frequency of detection, a limited and similar impact in both groups was observed for the most prevalent (over 80 %) periodontal pathogens (Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum). For proportions, reductions in P. gingivalis occurred at 3 months, both in the test and control groups (from 16.3 to 10 % and 16 to 14.8 %, respectively), although these differences were not statistically significant. At 12 months, the test group showed a statistically significant greater reduction in probing depth for the sampled sites. The adjunctive use of Er:YAG laser when compared with conventional ultrasonic debridement did not provide a microbiological added benefit. Even though some clinical benefits with the adjunctive laser application were identified when comparing both treatment protocols, there were no differences in microbiological outcomes or in the bacterial recolonization patterns.

  18. Subgingival bacteria in Ghanaian adolescents with or without progression of attachment loss

    Directory of Open Access Journals (Sweden)

    Gunnar Dahlén

    2014-05-01

    Full Text Available Objective: This study describes subgingival bacterial profiles associated with clinical periodontal status in Ghanaian adolescents with or without progression of attachment loss. Materials and methods: Among 500 adolescents included in a cohort study, 397 returned 2 years later for a periodontal re-examination, including full-mouth CAL measurements. At follow-up, a subgroup of 98 adolescents was also subjected to bacterial sampling with paper points at four periodontal sites (mesial aspect of 11, 26, 31, and 46 and analyzed with the checkerboard DNA–DNA hybridization technique against DNA-probes from nine periodontitis-associated bacterial species. Results: The 98 Ghanaian adolescents examined in the present study were similar to the entire group examined at the 2-year follow-up with respect to age, gender, and CAL ≥3 mm. A high detection frequency of Fusobacterium nucleatum and Prevotella intermedia (>99% using checkerboard analysis was found, while for Aggregatibacter actinomycetemcomitans the detection frequency was <50%. A strong correlation was found at the individual level between the presence of P. intermedia and the total CAL change, and P. intermedia and Porphyromonas gingivalis were strongly correlated with a change in CAL and probing pocket depth (PPD at the sampled sites. In a linear regression model, a significant discriminating factor for the total CAL change in the dentition during the 2-year follow-up period was obtained for P. intermedia and public school. Conclusion: This study indicates that subgingival bacterial species other than A. actinomycetemcomitans, for example, P. intermedia, have a significant association with periodontal breakdown (change in CAL in Ghanaian adolescents with progression of periodontal attachment loss.

  19. The same microbiota and a potentially discriminant metabolome in the saliva of omnivore, ovo-lacto-vegetarian and Vegan individuals.

    Directory of Open Access Journals (Sweden)

    Francesca De Filippis

    Full Text Available The salivary microbiota has been linked to both oral and non-oral diseases. Scant knowledge is available on the effect of environmental factors such as long-term dietary choices on the salivary microbiota and metabolome. This study analyzed the microbial diversity and metabolomic profiles of the saliva of 161 healthy individuals who followed an omnivore or ovo-lacto-vegetarian or vegan diet. A large core microbiota was identified, including 12 bacterial genera, found in >98% of the individuals. The subjects could be stratified into three "salivary types" that differed on the basis of the relative abundance of the core genera Prevotella, Streptococcus/Gemella and Fusobacterium/Neisseria. Statistical analysis indicated no effect of dietary habit on the salivary microbiota. Phylogenetic beta-diversity analysis consistently showed no differences between omnivore, ovo-lacto-vegetarian and vegan individuals. Metabolomic profiling of saliva using (1H-NMR and GC-MS/SPME identified diet-related biomarkers that enabled a significant discrimination between the 3 groups of individuals on the basis of their diet. Formate, urea, uridine and 5-methyl-3-hexanone could discriminate samples from omnivores, whereas 1-propanol, hexanoic acid and proline were characteristic of non-omnivore diets. Although the salivary metabolome can be discriminating for diet, the microbiota has a remarkable inter-individual stability and did not vary with dietary habits. Microbial homeostasis might be perturbed with sub-standard oral hygiene or other environmental factors, but there is no current indication that a choice of an omnivore, ovo-lacto-vegetarian or vegan diet can lead to a specific composition of the oral microbiota with consequences on the oral homeostasis.

  20. Identification of Candida dubliniensis in a study of HIV-seropositive pediatric dental patients.

    Science.gov (United States)

    Brown, D M; Jabra-Rizk, M A; Falkler, W A; Baqui, A A; Meiller, T F

    2000-01-01

    The combination of an immature immune system and suppressed cellular immunity in children with HIV infections provides optimal conditions for rapid disease progression. As a result, pediatric AIDS has become a major epidemiological challenge. Oral fungal colonization remains one of the most common opportunistic infections observed in both adult and pediatric HIV infected patients. Although Candida albicans is the most frequently isolated opportunistic fungal species, a recently characterized Candida species, C. dubliniensis, has gained considerable attention due to its almost exclusive association with HIV-seropositive individuals. The purpose of this study was to prospectively screen for the presence of C. dubliniensis among pediatric HIV+ patients. Oral samples taken from twenty-seven children were cultured for the presence of yeast. All positive yeast isolates obtained were screened for the presence of C. dubliniensis by use of tests for germ tube and chlamydospore production, detection of inability to grow at 45 degrees C, by colony color on CHROMagar Candida medium, coaggregation with Fusobacterium nucleatum ATCC 49256 and by the results of sugar assimilation testing with the API 20C AUX yeast identification system. Among the 27 patients tested, 3 patients were found to harbor C. dubliniensis, one of which also grew C. glabrata; 12 patients were colonized with C. albicans, while the remaining 12 patients were negative for yeast. Identification of the three C. dubliniensis isolates was genetically confirmed by electrophoretic karyotyping. All three C. dubliniensis isolates were found to be susceptible to fluconazole (MIC pediatric HIV seropositive population and support the need for further investigation into the prevalence and pathogenesis of C. dubliniensis.

  1. Gut microbiota, metabolome and immune signatures in patients with uncomplicated diverticular disease.

    Science.gov (United States)

    Barbara, Giovanni; Scaioli, Eleonora; Barbaro, Maria Raffaella; Biagi, Elena; Laghi, Luca; Cremon, Cesare; Marasco, Giovanni; Colecchia, Antonio; Picone, Gianfranco; Salfi, Nunzio; Capozzi, Francesco; Brigidi, Patrizia; Festi, Davide

    2017-07-01

    The engagement of the gut microbiota in the development of symptoms and complications of diverticular disease has been frequently hypothesised. Our aim was to explore colonic immunocytes, gut microbiota and the metabolome in patients with diverticular disease in a descriptive, cross-sectional, pilot study. Following colonoscopy with biopsy and questionnaire phenotyping, patients were classified into diverticulosis or symptomatic uncomplicated diverticular disease; asymptomatic subjects served as controls. Mucosal immunocytes, in the diverticular region and in unaffected sites, were quantified with immunohistochemistry. Mucosa and faecal microbiota were analysed by the phylogenetic platform high taxonomic fingerprint (HTF)-Microbi.Array, while the metabolome was assessed by 1 H nuclear magnetic resonance. Compared with controls, patients with diverticula, regardless of symptoms, had a >70% increase in colonic macrophages. Their faecal microbiota showed depletion of Clostridium cluster IV. Clostridium cluster IX, Fusobacterium and Lactobacillaceae were reduced in symptomatic versus asymptomatic patients. A negative correlation was found between macrophages and mucosal Clostridium cluster IV and Akkermansia . Urinary and faecal metabolome changes in diverticular disease involved the hippurate and kynurenine pathways. Six urinary molecules allowed to discriminate diverticular disease and control groups with >95% accuracy. Patients with colonic diverticular disease show depletion of microbiota members with anti-inflammatory activity associated with mucosal macrophage infiltration. Metabolome profiles were linked to inflammatory pathways and gut neuromotor dysfunction and showed the ability to discriminate diverticular subgroups and controls. These data pave the way for further large-scale studies specifically aimed at identifying microbiota signatures with a potential diagnostic value in patients with diverticular disease. Published by the BMJ Publishing Group Limited

  2. Analysis of potential risks from the bacterial communities associated with air-contact surfaces from tilapia (Oreochromis niloticus) fish farming.

    Science.gov (United States)

    Grande Burgos, Maria Jose; Romero, Jose Luis; Pérez Pulido, Rubén; Cobo Molinos, Antonio; Gálvez, Antonio; Lucas, Rosario

    2018-01-01

    Tilapia farming is a promising growing sector in aquaculture. Yet, there are limited studies on microbiological risks associated to tilapia farms. The aim of the present study was to analyse the bacterial communities from solid surfaces in contact with air in a tilapia farm in order to evaluate the presence of bacteria potentially toxinogenic or pathogenic to humans or animals. Samples from a local tilapia farm (tank wall, aerator, water outlets, sink and floor) were analyzed by high throughput sequencing technology. Sequences were assigned to operational taxonomic units (OTUs). Proteobacteria was the main phylum represented in most samples (except for one). Cyanobacteria were a relevant phylum in the inner wall from the fattening tank and the wet floor by the pre-fattening tank. Bacteroidetes were the second phylum in relative abundance for samples from the larval rearing tank and the pre-fattening tank and one sample from the fattening tank. Fusobacteria showed highest relative abundances in samples from the larval rearing tank and pre-fattening tank. Other phyla (Verrucomicrobia, Actinobacteria, Firmicutes, Planktomycetes, Acidobacteria, Chloroflexi, Chlorobi, Gemmatiomonadetes or Fibrobacters) had lower relative abundances. A large fraction of the reads (ranging from 43.67% to 72.25%) were assigned to uncultured bacteria. Genus Acinetobacter (mainly A. calcoaceticus/baumanni) was the predominant OTU in the aerator of the fattening tank and also in the nearby sink on the floor. The genera Cetobacterium and Bacteroides showed highest relative abundances in the samples from the larval rearing tank and the pre-fattening tank. Genera including fish pathogens (Fusobacterium, Aeromonas) were only detected at low relative abundances. Potential human pathogens other than Acinetobacter were either not detected or had very low relative abundances (Acinetobacter and potential cyanotoxin-producing cyanobacteria. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Immunohistochemical analysis of the gingiva with periodontitis of type I plasminogen deficiency compared to gingiva with gingivitis and periodontitis and healthy gingiva.

    Science.gov (United States)

    Kurtulus Waschulewski, Idil; Gökbuget, Aslan Y; Christiansen, Nina M; Ziegler, Maike; Schuster, Volker; Wahl, Gerhard; Götz, Werner

    2016-12-01

    Type I plasminogen deficiency (Plgdef) is an uncommon chronic inflammation of mucous membranes. Gingival enlargements usually proceed with progressive periodontal destruction and tooth-loss. Plasmin(ogen)-independent enzymatic mechanisms for fibrin clearance have already been discussed in the literature. Our primary objective was to verify, immunohistochemically, the occurrence of different enzymatic factors involved in tissue breakdown of inflamed compared to healthy gingiva. Secondly, we tried to find out, if these patients have a similar microbiological profile to the patients with known gingivitis and periodontitis. Immunohistochemical analysis of enzymes elastase, plasminogen (plg), cathepsin G, matrix-metalloproteinase (MMP)-3 and MMP-7 and of glycoprotein fibrinogen were performed with gingival tissues from 3 healthy controls, 8 patients with Plgdef and 3 patients with gingivitis and periodontitis. Furthermore, plaque from 5 patients with plasminogen deficiency were also obtained to determine the microbiological profile. Significantly high numbers of elastase positive leukocytes were detected in all samples. Staining for MMP-3 and MMP-7 was seen in samples with gingivitis and periodontitis with a stronger staining in samples with periodontitis by Plgdef. Fibrinogen was detectable in all samples. Staining for plg was stronger in samples with periodontitis than in other samples. Staining for cathepsin G was weak in gingivitis and periodontitis. Subgingival microbial flora showed elevated colony forming units of Prevotella intermedia/nigrescens, Fusobacterium spp., Eikenella corrodens, Porphyromonas gingivalis and viridans streptococci. Strong staining of elastase, MMP-3 and MMP-7 and weak staining of plg in Plgdef samples supports the plasmin(ogen) - independent fibrin clearance. Similar subgingival microbiological flora was observed in periodontitis with Plgdef as in other periodontal diseases. Further investigations should determine the exact pathomechanism

  4. O tabagismo como fator de risco para as doenças periodontais: aspectos microbiológicos

    Directory of Open Access Journals (Sweden)

    GAETTI-JARDIM JÚNIOR Elerson

    1998-01-01

    Full Text Available O fumo é considerado importante fator predisponente para muitas doenças, incluindo-se as periodontopatias. Desde que as doenças periodontais representam a inter-relação entre os fatores de virulência da microbiota subgengival sobre um hospedeiro susceptível, foi objetivo avaliar a freqüência de isolamento de três periodontopatógenos em indivíduos sadios e pacientes com doença periodontal, fumantes ou não, com níveis variados de higiene bucal; verificar a relação entre o número de microrganismos produtores de sulfeto de hidrogênio na placa subgengival de fumantes e não fumantes e sua condição clínica. Foram examinados 189 pacientes e indivíduos sadios, dos quais 60 foram selecionados para análise microbiológica. O índice de placa foi registrado de acordo com o índice de O'Leary e os espécimes de placa subgengival coletados e processados de acordo com SLOTS35 (1982. A identificação dos isolados foi obtida pelas suas características morfocelulares, morfocoloniais e bioquímico-fisiológicas. Verificou-se que a freqüência de isolamento dos bastonetes anaeróbios produtores de pigmento negro, Fusobacterium nucleatum e bactérias produtoras de sulfeto de hidrogênio foi similar entre fumantes e não fumantes, sendo mais elevada nos pacientes com doença periodontal. Já Actinobacillus actinomycetemcomitans foi isolado mais freqüentemente em sadios fumantes do que sadios não fumantes.

  5. Adhesion of periodontal pathogens to self-ligating orthodontic brackets: An in-vivo prospective study.

    Science.gov (United States)

    Jung, Woo-Sun; Kim, Kyungsun; Cho, Soha; Ahn, Sug-Joon

    2016-09-01

    Our aims were to analyze adhesion of periodontopathogens to self-ligating brackets (Clarity-SL [CSL], Clippy-C [CC] and Damon Q [DQ]) and to identify the relationships between bacterial adhesion and oral hygiene indexes. Central incisor brackets from the maxilla and mandible were collected from 60 patients at debonding after the plaque and gingival indexes were measured. Adhesions of Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), and Tannerella forsythia (Tf) were quantitatively determined using real-time polymerase chain reactions. Factorial analysis of variance was used to analyze bacterial adhesion in relation to bracket type and jaw position. Correlation coefficients were calculated to determine the relationships between bacterial adhesion and the oral hygiene indexes. Total bacteria showed greater adhesion to CSL than to DQ brackets, whereas Aa, Pg, and Pi adhered more to DQ than to CSL brackets. CC brackets showed an intermediate adhesion pattern between CSL and DQ brackets, but it did not differ significantly from either bracket type. Adhesion of Fn and Tf did not differ significantly among the 3 brackets. Most bacteria were detected in greater quantities in the mandibular than in the maxillary brackets. The plaque and gingival indexes were not strongly correlated with bacterial adhesion to the brackets. Because Aa, Pg, and Pi adhered more to the DQ brackets in the mandibular area, orthodontic patients with periodontal problems should be carefully monitored in the mandibular incisors where the distance between the bracket and the gingiva is small, especially when DQ brackets are used. Copyright © 2016 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.

  6. Microbiological, immunological and genetic factors in family members with periodontitis as a manifestation of systemic disease, associated with hematological disorders.

    Science.gov (United States)

    Okada, Mitsugi; Awane, Saori; Suzuki, Junji; Hino, Takamune; Takemoto, Toshinobu; Kurihara, Hidemi; Miura, Kazuo

    2002-08-01

    The microflora, immunological profiles of host defence functions, and human leukocyte antigen (HLA) findings are reported for a mother, son and daughter who were diagnosed as having 'periodontitis as a manifestation of systemic diseases, associated with hematological disorders'. Examinations were made of the bacterial flora from the periodontal pocket, neutrophil chemotaxis, neutrophil phagocytosis, and the genotypes (DQB1) and serotypes (DR locus) of HLA class II antigens. Phenotypic analyses of the peripheral lymphocytes were also conducted. The subgingival microflora from the mother was dominated by Gram-negative rods, especially Porphyromonas endodontalis, Prevotella intermedia/Prevotella nigrescens and Fusobacterium nucleatum. Subgingival microflora samples from the son and daughter were dominated by Gram-positive cocci and Gram-positive rods. Through the use of polymerase chain reaction, Campylobacter rectus and Capnocytophaga gingivalis were detected in all subjects, whereas Porphyromonas gingivalis, P. intermedia, and Treponema denticola were not detected in any subjects. All three subjects showed a remarkable level of depressed neutrophil chemotaxis to N-formyl-methionyl-leucyl-phenylalanine, although their phagocyte function levels were normal, in comparison to healthy control subjects. Each subject had the same genotype, HLA-DQB1*0601, while the mother had HLA-DR2 and HLA-DR8, and the son and daughter had HLA-DR2 only. In summary, the members of this family showed a similar predisposition to periodontitis with regard to certain host defence functions. It is suggested that the depressed neutrophil chemotaxis that was identified here could be a significant risk factor for periodontitis in this family.

  7. High-Level Antimicrobial Efficacy of Representative Mediterranean Natural Plant Extracts against Oral Microorganisms

    Directory of Open Access Journals (Sweden)

    Lamprini Karygianni

    2014-01-01

    Full Text Available Nature is an unexplored reservoir of novel phytopharmaceuticals. Since biofilm-related oral diseases often correlate with antibiotic resistance, plant-derived antimicrobial agents could enhance existing treatment options. Therefore, the rationale of the present report was to examine the antimicrobial impact of Mediterranean natural extracts on oral microorganisms. Five different extracts from Olea europaea, mastic gum, and Inula viscosa were tested against ten bacteria and one Candida albicans strain. The extraction protocols were conducted according to established experimental procedures. Two antimicrobial assays—the minimum inhibitory concentration (MIC assay and the minimum bactericidal concentration (MBC assay—were applied. The screened extracts were found to be active against each of the tested microorganisms. O. europaea presented MIC and MBC ranges of 0.07–10.00 mg mL−1 and 0.60–10.00 mg mL−1, respectively. The mean MBC values for mastic gum and I. viscosa were 0.07–10.00 mg mL−1 and 0.15–10.00 mg mL−1, respectively. Extracts were less effective against C. albicans and exerted bactericidal effects at a concentration range of 0.07–5.00 mg mL−1 on strict anaerobic bacteria (Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Parvimonas micra. Ethyl acetate I. viscosa extract and total mastic extract showed considerable antimicrobial activity against oral microorganisms and could therefore be considered as alternative natural anti-infectious agents.

  8. Comparing the bacterial diversity of acute and chronic dental root canal infections.

    Directory of Open Access Journals (Sweden)

    Adriana L Santos

    Full Text Available This study performed barcoded multiplex pyrosequencing with a 454 FLX instrument to compare the microbiota of dental root canal infections associated with acute (symptomatic or chronic (asymptomatic apical periodontitis. Analysis of samples from 9 acute abscesses and 8 chronic infections yielded partial 16S rRNA gene sequences that were taxonomically classified into 916 bacterial species-level operational taxonomic units (OTUs (at 3% divergence belonging to 67 genera and 13 phyla. The most abundant phyla in acute infections were Firmicutes (52%, Fusobacteria (17% and Bacteroidetes (13%, while in chronic infections the dominant were Firmicutes (59%, Bacteroidetes (14% and Actinobacteria (10%. Members of Fusobacteria were much more prevalent in acute (89% than in chronic cases (50%. The most abundant/prevalent genera in acute infections were Fusobacterium and Parvimonas. Twenty genera were exclusively detected in acute infections and 18 in chronic infections. Only 18% (n = 165 of the OTUs at 3% divergence were shared by acute and chronic infections. Diversity and richness estimators revealed that acute infections were significantly more diverse than chronic infections. Although a high interindividual variation in bacterial communities was observed, many samples tended to group together according to the type of infection (acute or chronic. This study is one of the most comprehensive in-deep comparisons of the microbiota associated with acute and chronic dental root canal infections and highlights the role of diverse polymicrobial communities as the unit of pathogenicity in acute infections. The overall diversity of endodontic infections as revealed by the pyrosequencing technique was much higher than previously reported for endodontic infections.

  9. Frequency and levels of candidate endodontic pathogens in acute apical abscesses as compared to asymptomatic apical periodontitis.

    Science.gov (United States)

    Rôças, Isabela N; Siqueira, José F

    2018-01-01

    Acute apical abscess is caused by bacteria that leave the infected dental root canal to invade the periodontal tissues. Most species occurring in abscesses are also found in asymptomatic infections; therefore, the possibility exists that not only the presence of certain species but also their specific counts influence the appearance of symptoms. This molecular study compared the frequency and levels of several candidate endodontic pathogens in teeth with acute apical abscesses and asymptomatic apical periodontitis. Samples were taken from the root canals of teeth with asymptomatic apical periodontitis (n = 73) and by aspiration of purulent exudate from acute abscesses (n = 55). DNA was extracted from samples and bacterial identifications were performed by a closed-ended semi-quantitative reverse-capture checkerboard approach targeting 40 bacterial species/phylotypes. Bacterial DNA was detected in all cases. In abscesses, the most prevalent taxa were Fusobacterium nucleatum (60%), Porphyromonas endodontalis (53%), Parvimonas micra (51%), and Streptococcus species (45%). The most frequently detected taxa in asymptomatic teeth were P. endodontalis (63%), Dialister invisus (58%), Olsenella uli (56%), and F. nucleatum (51%). None of the targeted taxa were significantly associated with abscesses when only mere presence was evaluated (p>0.05). However, semi-quantitative data demonstrated that P. endodontalis, Prevotella baroniae, Treponema denticola and Streptococcus species were significantly more frequent at levels >105 in abscesses than in asymptomatic cases (p<0.05). None of the target species/phylotypes were associated with abscesses in terms of frequency. However, some taxa were significantly found in higher levels in abscesses. Presence of a potentially virulent pathogen in high counts may increase the collective pathogenicity of the bacterial community and give rise to symptoms.

  10. Comparison of endotoxin levels found in primary and secondary endodontic infections.

    Science.gov (United States)

    Gomes, Brenda P F A; Endo, Marcos S; Martinho, Frederico C

    2012-08-01

    This clinical study was conducted to compare the levels of endotoxins (lipopolysaccharides [LPSs]) found in primary and secondary endodontic infections with apical periodontitis by correlating LPS contents with clinical/radiographic findings. In addition, the presence of target gram-negative anaerobic bacteria was also investigated. Samples were taken from 15 root canals with primary infections and 15 with secondary infections by using paper points. The limulus amebocyte lysate assay was used to quantify endotoxins, and the polymerase chain reaction technique (16S rDNA) was used for bacterial investigation. Endotoxins were detected in 100% of the root canal samples collected from primary (15/15) and secondary (15/15) infections with median values of 7.49 EU/mL and 3.96 EU/mL, respectively (P asymptomatic teeth with primary infections (P 3 mm) (P < .05). Prevotella nigrescens (10/15, 4/15), Fusobacterium nucleatum (5/15, 1/15), Treponema denticola (3/15, 1/15), and Treponema socranskii (5/15, 1/15) were detected in teeth with primary and secondary infections, respectively. P. endodontalis was present only in teeth with primary infections (5/15). Teeth with primary endodontic infections had higher contents of endotoxins and a more complex gram-negative bacterial community than teeth with secondary infections. Moreover, the levels of endotoxins were related to the severity of bone destruction in periapical tissues as well as the development of clinical features in teeth with primary infections. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  11. Analysis of symptomatic and asymptomatic primary root canal infections in adult Norwegian patients.

    Science.gov (United States)

    Rôças, Isabela N; Siqueira, José F; Debelian, Gilberto J

    2011-09-01

    This molecular study analyzed the microbiota of primary root canal infections from adult Norwegian patients. Samples were taken from the necrotic root canals of teeth with symptomatic (n = 13) or asymptomatic (n = 21) apical periodontitis and chronic apical abscesses (n = 9). DNA was extracted from samples, and bacterial identifications were performed by a closed-ended reverse-capture checkerboard approach targeting 50 candidate endodontic pathogens. Bacterial DNA was detected in all cases. In teeth with asymptomatic apical periodontitis, the most frequent taxa were Dialister invisus (71%), Fusobacterium nucleatum (62%), and Porphyromonas endodontalis (62%). In chronic apical abscesses, the most prevalent taxa were P. endodontalis (100%), D. invisus (89%), Parvimonas micra (78%), and Solobacterium moorei (78%). In teeth with symptomatic apical periodontitis, the most prevalent taxa were D. invisus, P. endodontalis, S. moorei, Propionibacterium acnes, and Streptococcus species (all in 69%). None of the targeted taxa were significantly associated with either sinus tract or pain (P > .05), except for Selenomonas sputigena, which was more frequently found in painful cases (P = .04). No taxa were found in significantly higher levels in any conditions (P > .05). Cluster analyses revealed bacterial groupings that differed between cases with and without pain. Although basically the same species were highly prevalent in the different conditions examined and none of the most prevalent taxa were positively associated with symptoms, results revealed that species formed different partnerships and associations in samples from teeth with or without pain. Therefore, it is possible that more virulent multispecies communities can form as a result of overall bacterial combinations and give rise to acute inflammation. Copyright © 2011 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  12. Microbial community in persistent apical periodontitis: a 16S rRNA gene clone library analysis.

    Science.gov (United States)

    Zakaria, M N; Takeshita, T; Shibata, Y; Maeda, H; Wada, N; Akamine, A; Yamashita, Y

    2015-08-01

    To characterize the microbial composition of persistent periapical lesions of root filled teeth using a molecular genetics approach. Apical lesion samples were collected from 12 patients (23-80 years old) who visited the Kyushu University Hospital for apicectomy with persistent periapical lesions associated with root filled teeth. DNA was directly extracted from each sample and the microbial composition was comprehensively analysed using clone library analysis of the 16S rRNA gene. Enterococcus faecalis, Candida albicans and specific fimA genotypes of Porphyromonas gingivalis were confirmed using polymerase chain reaction (PCR) analysis with specific primers. Bacteria were detected in all samples, and the dominant findings were P. gingivalis (19.9%), Fusobacterium nucleatum (11.2%) and Propionibacterium acnes (9%). Bacterial diversity was greater in symptomatic lesions than in asymptomatic ones. In addition, the following bacteria or bacterial combinations were characteristic to symptomatic lesions: Prevotella spp., Treponema spp., Peptostreptococcaceae sp. HOT-113, Olsenella uli, Slackia exigua, Selemonas infelix, P. gingivalis with type IV fimA, and a combination of P. gingivalis, F. nucleatum, and Peptostreptococcaceae sp. HOT-113 and predominance of Streptococcus spp. On the other hand, neither Enterococcus faecalis nor C. albicans were detected in any of the samples. Whilst a diverse bacterial species were observed in the persistent apical lesions, some characteristic patterns of bacterial community were found in the symptomatic lesions. The diverse variation of community indicates that bacterial combinations as a community may cause persistent inflammation in periapical tissues rather than specific bacterial species. © 2014 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  13. Two FOXP3(+)CD4(+) T cell subpopulations distinctly control the prognosis of colorectal cancers.

    Science.gov (United States)

    Saito, Takuro; Nishikawa, Hiroyoshi; Wada, Hisashi; Nagano, Yuji; Sugiyama, Daisuke; Atarashi, Koji; Maeda, Yuka; Hamaguchi, Masahide; Ohkura, Naganari; Sato, Eiichi; Nagase, Hirotsugu; Nishimura, Junichi; Yamamoto, Hirofumi; Takiguchi, Shuji; Tanoue, Takeshi; Suda, Wataru; Morita, Hidetoshi; Hattori, Masahira; Honda, Kenya; Mori, Masaki; Doki, Yuichiro; Sakaguchi, Shimon

    2016-06-01

    CD4(+) T cells that express the forkhead box P3 (FOXP3) transcription factor function as regulatory T (Treg) cells and hinder effective immune responses against cancer cells. Abundant Treg cell infiltration into tumors is associated with poor clinical outcomes in various types of cancers. However, the role of Treg cells is controversial in colorectal cancers (CRCs), in which FOXP3(+) T cell infiltration indicated better prognosis in some studies. Here we show that CRCs, which are commonly infiltrated by suppression-competent FOXP3(hi) Treg cells, can be classified into two types by the degree of additional infiltration of FOXP3(lo) nonsuppressive T cells. The latter, which are distinguished from FOXP3(+) Treg cells by non-expression of the naive T cell marker CD45RA and instability of FOXP3, secreted inflammatory cytokines. Indeed, CRCs with abundant infiltration of FOXP3(lo) T cells showed significantly better prognosis than those with predominantly FOXP3(hi) Treg cell infiltration. Development of such inflammatory FOXP3(lo) non-Treg cells may depend on secretion of interleukin (IL)-12 and transforming growth factor (TGF)-β by tissues and their presence was correlated with tumor invasion by intestinal bacteria, especially Fusobacterium nucleatum. Thus, functionally distinct subpopulations of tumor-infiltrating FOXP3(+) T cells contribute in opposing ways to determining CRC prognosis. Depletion of FOXP3(hi) Treg cells from tumor tissues, which would augment antitumor immunity, could thus be used as an effective treatment strategy for CRCs and other cancers, whereas strategies that locally increase the population of FOXP3(lo) non-Treg cells could be used to suppress or prevent tumor formation.

  14. Alterations in oral microbial flora induced by waterpipe tobacco smoking

    Directory of Open Access Journals (Sweden)

    Shakhatreh MAK

    2018-02-01

    Full Text Available Muhamad Ali K Shakhatreh,1 Omar F Khabour,1 Karem H Alzoubi,2 Majed M Masadeh,3 Emad I Hussein,4 George N Bshara1 1Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, Irbid, Jordan; 2Department of Clinical Pharmacy, Faculty of Pharmacy, Jordan University of Science and Technology, Irbid, Jordan; 3Department of Pharmaceutical Technology, Faculty of Pharmacy, Jordan University of Science and Technology, Irbid, Jordan; 4Department of Biological Sciences, Yarmouk University, Irbid, Jordan Background: Waterpipe smoking is a global health problem and a serious public concern. Little is known about the effects of waterpipe smoking on oral health. In the current study, we examined the alterations of oral microbial flora by waterpipe smoking. Methods: One hundred adult healthy subjects (59 waterpipe smokers and 41 non-smokers were recruited into the study. Swabs were taken from the oral cavity and subgingival regions. Standard culturing techniques were used to identify types, frequency, and mean number of microorganisms in cultures obtained from the subjects. Results: It was notable that waterpipe smokers were significantly associated with a history of oral infections. In subgingiva, Acinetobacter and Moraxella species were present only in waterpipe smokers. In addition, the frequency of Candida albicans was higher in the subgingiva of waterpipe smokers (p = 0.023 while the frequency of Fusobacterium nucleatum was significantly lower in the subgingiva of waterpipe smokers (p = 0.036. However, no change was observed in other tested bacteria, such as Campylobacter species; Viridans group streptococci, Enterobacteriaceae, and Staphylococcus aureus. In oral cavity and when colony-forming units were considered, the only bacterial species that showed significant difference were the black-pigmented bacteria (p < 0.001. Conclusion: This study provides evidence indicating that some of the oral microflora is significantly altered by

  15. Interspecies dynamics among bacteria associated with canine periodontal disease.

    Science.gov (United States)

    Sanguansermsri, P; Nobbs, A H; Jenkinson, H F; Surarit, R

    2018-02-01

    The etiology and pathogenic mechanisms associated with canine periodontal disease are less well understood than the disease in humans. In this study we have reconstructed defined consortia biofilms in vitro of microorganisms identified as prevalent in a same-breed cohort of dogs with or without periodontal disease. Frederiksenia canicola and Neisseria canis were selected as potential early colonizers of salivary pellicle, and Fusobacterium nucleatum and Porphyromonas gulae were included as high incidence canine oral bacteria. N. canis formed a biofilm substratum under aerobic conditions, but was unable to tolerate anaerobic conditions. Fr. canicola exhibited synergistic biofilm growth with Po. gulae under anaerobic conditions, but displayed an antagonistic relationship with Fu. nucleatum. However, strong co-adhesion between Fu. nucleatum and Po. gulae was able to overcome the inhibitory effects of Fr. canicola to facilitate three-species biofilm formation. Parvimonas micra, an anaerobic, asaccharolytic Gram-positive coccus found only under disease conditions in vivo, was able to form biofilms in conjunction with Fr. canicola and Po. gulae. Furthermore, the specific proteolytic activities of biofilms containing Fr. canicola and Po. gulae or Fu. nucleatum and Po. gulae were increased several-fold upon the addition of Pa. micra. This suggests that anaerobic cocci such as Pa. micra might provide a catalyst for progressive tissue destruction, inflammation and alveolar bone loss in canine periodontal disease, in keeping with the keystone-pathogen hypothesis. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  16. Oral bacterial DNA findings in pericardial fluid

    Directory of Open Access Journals (Sweden)

    Anne-Mari Louhelainen

    2014-11-01

    Full Text Available Background: We recently reported that large amounts of oral bacterial DNA can be found in thrombus aspirates of myocardial infarction patients. Some case reports describe bacterial findings in pericardial fluid, mostly done with conventional culturing and a few with PCR; in purulent pericarditis, nevertheless, bacterial PCR has not been used as a diagnostic method before. Objective: To find out whether bacterial DNA can be measured in the pericardial fluid and if it correlates with pathologic–anatomic findings linked to cardiovascular diseases. Methods: Twenty-two pericardial aspirates were collected aseptically prior to forensic autopsy at Tampere University Hospital during 2009–2010. Of the autopsies, 10 (45.5% were free of coronary artery disease (CAD, 7 (31.8% had mild and 5 (22.7% had severe CAD. Bacterial DNA amounts were determined using real-time quantitative PCR with specific primers and probes for all bacterial strains associated with endodontic disease (Streptococcus mitis group, Streptococcus anginosus group, Staphylococcus aureus/Staphylococcus epidermidis, Prevotella intermedia, Parvimonas micra and periodontal disease (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Fusobacterium nucleatus, and Dialister pneumosintes. Results: Of 22 cases, 14 (63.6% were positive for endodontic and 8 (36.4% for periodontal-disease-associated bacteria. Only one case was positive for bacterial culturing. There was a statistically significant association between the relative amount of bacterial DNA in the pericardial fluid and the severity of CAD (p=0.035. Conclusions: Oral bacterial DNA was detectable in pericardial fluid and an association between the severity of CAD and the total amount of bacterial DNA in pericardial fluid was found, suggesting that this kind of measurement might be useful for clinical purposes.

  17. Diets Alter the Gut Microbiome of Crocodile Lizards

    Directory of Open Access Journals (Sweden)

    Hai-Ying Jiang

    2017-10-01

    Full Text Available The crocodile lizard is a critically endangered reptile, and serious diseases have been found in this species in recent years, especially in captive lizards. Whether these diseases are caused by changes in the gut microbiota and the effect of captivity on disease remains to be determined. Here, we examined the relationship between the gut microbiota and diet and disease by comparing the fecal microbiota of wild lizards with those of sick and healthy lizards in captivity. The gut microbiota in wild crocodile lizards was consistently dominated by Proteobacteria (∼56.4% and Bacteroidetes (∼19.1%. However, the abundance of Firmicutes (∼2.6% in the intestine of the wild crocodile lizards was distinctly lower than that in other vertebrates. In addition, the wild samples from Guangdong Luokeng Shinisaurus crocodilurus National Nature Reserve also had a high abundance of Deinococcus–Thermus while the wild samples from Guangxi Daguishan Crocodile Lizard National Nature Reserve had a high abundance of Tenericutes. The gut microbial community in loach-fed crocodile lizards was significantly different from the gut microbial community in the earthworm-fed and wild lizards. In addition, significant differences in specific bacteria were detected among groups. Notably, in the gut microbiota, the captive lizards fed earthworms resulted in enrichment of Fusobacterium, and the captive lizards fed loaches had higher abundances of Elizabethkingia, Halomonas, Morganella, and Salmonella, all of which are pathogens or opportunistic pathogens in human or other animals. However, there is no sufficient evidence that the gut microbiota contributes to either disease A or disease B. These results provide a reference for the conservation of endangered crocodile lizards and the first insight into the relationship between disease and the gut microbiota in lizards.

  18. Early colonizing Escherichia coli elicits remodeling of rat colonic epithelium shifting toward a new homeostatic state.

    Science.gov (United States)

    Tomas, Julie; Reygner, Julie; Mayeur, Camille; Ducroc, Robert; Bouet, Stephan; Bridonneau, Chantal; Cavin, Jean-Baptiste; Thomas, Muriel; Langella, Philippe; Cherbuy, Claire

    2015-01-01

    We investigated the effects of early colonizing bacteria on the colonic epithelium. We isolated dominant bacteria, Escherichia coli, Enterococcus faecalis, Lactobacillus intestinalis, Clostridium innocuum and a novel Fusobacterium spp., from the intestinal contents of conventional suckling rats and transferred them in different combinations into germfree (GF) adult rats. Animals were investigated after various times up to 21 days. Proliferative cell markers (Ki67, proliferating cell nuclear antigen, phospho-histone H3, cyclin A) were higher in rats monocolonized with E. coli than in GF at all time points, but not in rats monocolonized with E. faecalis. The mucin content of goblet cells declined shortly after E. coli administration whereas the mucus layer doubled in thickness. Fluorescence in situ hybridization analyses revealed that E. coli resides in this mucus layer. The epithelial mucin content progressively returned to baseline, following an increase in KLF4 and in the cell cycle arrest-related proteins p21(CIP1) and p27(KIP1). Markers of colonic differentiated cells involved in electrolyte (carbonic anhydrase II and slc26A3) and water (aquaglyceroporin3 (aqp3)) transport, and secretory responses to carbachol were modulated after E. coli inoculation suggesting that ion transport dynamics were also affected. The colonic responses to simplified microbiotas differed substantially according to whether or not E. coli was combined with the other four bacteria. Thus, proliferation markers increased substantially when E. coli was in the mix, but very much less when it was absent. This work demonstrates that a pioneer strain of E. coli elicits sequential epithelial remodeling affecting the structure, mucus layer and ionic movements and suggests this can result in a microbiota-compliant state.

  19. Microbiota present in cystic fibrosis lungs as revealed by whole genome sequencing.

    Directory of Open Access Journals (Sweden)

    Philippe M Hauser

    Full Text Available Determination of the precise composition and variation of microbiota in cystic fibrosis lungs is crucial since chronic inflammation due to microorganisms leads to lung damage and ultimately, death. However, this constitutes a major technical challenge. Culturing of microorganisms does not provide a complete representation of a microbiota, even when using culturomics (high-throughput culture. So far, only PCR-based metagenomics have been investigated. However, these methods are biased towards certain microbial groups, and suffer from uncertain quantification of the different microbial domains. We have explored whole genome sequencing (WGS using the Illumina high-throughput technology applied directly to DNA extracted from sputa obtained from two cystic fibrosis patients. To detect all microorganism groups, we used four procedures for DNA extraction, each with a different lysis protocol. We avoided biases due to whole DNA amplification thanks to the high efficiency of current Illumina technology. Phylogenomic classification of the reads by three different methods produced similar results. Our results suggest that WGS provides, in a single analysis, a better qualitative and quantitative assessment of microbiota compositions than cultures and PCRs. WGS identified a high quantity of Haemophilus spp. (patient 1 or Staphylococcus spp. plus Streptococcus spp. (patient 2 together with low amounts of anaerobic (Veillonella, Prevotella, Fusobacterium and aerobic bacteria (Gemella, Moraxella, Granulicatella. WGS suggested that fungal members represented very low proportions of the microbiota, which were detected by cultures and PCRs because of their selectivity. The future increase of reads' sizes and decrease in cost should ensure the usefulness of WGS for the characterisation of microbiota.

  20. Analysis of bacterial flora associated with peri-implantitis using obligate anaerobic culture technique and 16S rDNA gene sequence.

    Science.gov (United States)

    Tamura, Naoki; Ochi, Morio; Miyakawa, Hiroshi; Nakazawa, Futoshi

    2013-01-01

    To analyze and characterize the predominant bacterial flora associated with peri-implantitis by using culture techniques under obligate anaerobic conditions and 16S rDNA gene sequences. Subgingival bacterial specimens were taken from 30 patients: control (n = 15), consisting of patients with only healthy implants; and test (n = 15), consisting of patients with peri-implantitis. In both groups, subgingival bacterial specimens were taken from the deepest sites. An anaerobic glove box system was used to cultivate bacterial strains. The bacterial strains were identified by 16S rDNA genebased polymerase chain reaction and comparison of the gene sequences. Peri-implantitis sites had approximately 10-fold higher mean colony forming units (per milliliter) than healthy implant sites. A total of 69 different bacterial species were identified in the peri-implantitis sites and 53 in the healthy implant sites. The predominant bacterial species in the peri-implantitis sites were Eubacterium nodatum, E. brachy, E. saphenum, Filifactor alocis, Slackia exigua, Parascardovia denticolens, Prevotella intermedia, Fusobacterium nucleatum, Porphyromonas gingivalis, Centipeda periodontii, and Parvimonas micra. The predominant bacteria in healthy implant sites apart from Streptococcus were Pseudoramibacter alactolyticus, Veillonella species, Actinomyces israelii, Actinomyces species, Propionibacterium acnes, and Parvimonas micra. These results suggest that the environment in the depths of the sulcus showing peri-implantitis is well suited for growth of obligate anaerobic bacteria. The present study demonstrated that the sulcus around oral implants with peri-implantitis harbors high levels of asaccharolytic anaerobic gram-positive rods (AAGPRs) such as E. nodatum, E. brachy, E. saphenum, Filifactor alocis, Slackia exigua, and gram-negative anaerobic rods, suggesting that conventional periodontopathic bacteria are not the only periodontal pathogens active in peri-implantitis, and that AAGPRs

  1. Multi-centre evaluation of mass spectrometric identification of anaerobic bacteria using the VITEK® MS system.

    Science.gov (United States)

    Garner, O; Mochon, A; Branda, J; Burnham, C-A; Bythrow, M; Ferraro, M; Ginocchio, C; Jennemann, R; Manji, R; Procop, G W; Richter, S; Rychert, J; Sercia, L; Westblade, L; Lewinski, M

    2014-04-01

    Accurate and timely identification of anaerobic bacteria is critical to successful treatment. Classic phenotypic methods for identification require long turnaround times and can exhibit poor species level identification. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is an identification method that can provide rapid identification of anaerobes. We present a multi-centre study assessing the clinical performance of the VITEK(®) MS in the identification of anaerobic bacteria. Five different test sites analysed a collection of 651 unique anaerobic isolates comprising 11 different genera. Multiple species were included for several of the genera. Briefly, anaerobic isolates were applied directly to a well of a target plate. Matrix solution (α-cyano-4-hydroxycinnamic acid) was added and allowed to dry. Mass spectra results were generated with the VITEK(®) MS, and the comparative spectral analysis and organism identification were determined using the VITEK(®) MS database 2.0. Results were confirmed by 16S rRNA gene sequencing. Of the 651 isolates analysed, 91.2% (594/651) exhibited the correct species identification. An additional eight isolates were correctly identified to genus level, raising the rate of identification to 92.5%. Genus-level identification consisted of Actinomyces, Bacteroides and Prevotella species. Fusobacterium nucleatum, Actinomyces neuii and Bacteroides uniformis were notable for an increased percentage of no-identification results compared with the other anaerobes tested. VITEK(®) MS identification of clinically relevant anaerobes is highly accurate and represents a dramatic improvement over other phenotypic methods in accuracy and turnaround time. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

  2. Impact of supragingival therapy on subgingival microbial profile in smokers versus non-smokers with severe chronic periodontitis

    Directory of Open Access Journals (Sweden)

    Tatiana Meulman

    2012-01-01

    Full Text Available The aim of this study was to assess subgingival microbiological changes in smokers versus non-smokers presenting severe chronic periodontitis after supragingival periodontal therapy (ST.Non-smokers (n=10 and smokers (n=10 presenting at least nine teeth with probing pocket depth (PPD (≥5 mm, bleeding on probing (BoP, and no history of periodontal treatment in the last 6 months were selected. Clinical parameters assessed were plaque index (PI, BoP, PPD, relative gingival margin position (rGMP and relative clinical attachment level (rCAL. Subgingival biofilm was collected before and 21 days after ST. DNA was extracted and the 16S rRNA gene was amplified with the universal primer pair, 27F and 1492R. Amplified genes were cloned, sequenced, and identified by comparison with known 16S rRNA sequences. Statistical analysis was performed by Student's t and Chi-Square tests (α=5%.Clinically, ST promoted a significant reduction in PI and PPD, and gain of rCAL for both groups, with no significant intergroup difference. Microbiologically, at baseline, data analysis demonstrated that smokers harbored a higher proportion of Porphyromonas endodontalis, Bacteroidetes sp., Fusobacterium sp. and Tannerella forsythia and a lower number of cultivated phylotypes (p<0.05. Furthermore, non-smokers featured significant reductions in key phylotypes associated with periodontitis, whereas smokers presented more modest changes.Within the limits of the present study, ST promoted comparable clinical improvements in smokers and non-smokers with severe chronic periodontitis. However, in smokers, ST only slightly affected the subgingival biofilm biodiversity, as compared with non-smokers.

  3. Immunologic burden links periodontitis to acute coronary syndrome.

    Science.gov (United States)

    Liljestrand, John M; Paju, Susanna; Pietiäinen, Milla; Buhlin, Kåre; Persson, G Rutger; Nieminen, Markku S; Sinisalo, Juha; Mäntylä, Päivi; Pussinen, Pirkko J

    2018-01-01

    Periodontitis, a common polymicrobial inflammatory disease in the tooth supporting tissues, is a risk factor for coronary artery disease. One of the proposed underlying mechanisms is the systemic immune response to periodontal infection. We studied how serum antibodies against seven periodontal pathogens and their subgingival levels associate with each other, periodontitis, and coronary artery disease. The Parogene cohort included 505 Finnish patients (mean age 63 y) who underwent coronary angiography, and clinical and radiographic oral examinations. Coronary diagnosis was defined as no significant coronary artery disease (disease (≥50% stenosis, n = 184) and acute coronary syndrome (n = 169). Levels of subgingival Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, Tannerella forsythia, Campylobacter rectus, and Fusobacterium nucleatum were determined by checkerboard DNA-DNA hybridization. Serum antibody (IgA/IgG) levels were analyzed with enzyme-linked immunosorbent assay (ELISA). Aggregate IgA/IgG burdens were calculated by summing and standardizing the serum antibody levels. Patients with active periodontitis were characterized by higher levels of subgingival bacteria and corresponding IgA/IgG response. Quartiles 2-4 of serum IgA/IgG burden indicated higher risk for acute coronary syndrome (OR 1.84, 95%CI 1.01-3.35 for IgA; OR 1.87, 95%CI 1.01-3.46 for IgG) independently of established cardiovascular risk factors, body mass index, number of teeth, subgingival bacterial levels and periodontal diagnosis. Our findings support the hypothesis that the association between periodontitis and cardiovascular diseases is partly mediated by the immunologic response for periodontal pathogens. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Oral microbiota species in acute apical endodontic abscesses

    Directory of Open Access Journals (Sweden)

    Noelle George

    2016-03-01

    Full Text Available Background and objectives: Acute apical abscesses are serious endodontic diseases resulting from pulpal infection with opportunistic oral microorganisms. The objective of this study was to identify and compare the oral microbiota in patients (N=18 exhibiting acute apical abscesses, originating from the demographic region in Portland, Oregon. The study hypothesis is that abscesses obtained from this demographic region may contain unique microorganisms not identified in specimens from other regions. Design: Endodontic abscesses were sampled from patients at the Oregon Health & Science University (OHSU School of Dentistry. DNA from abscess specimens was subjected to polymerase chain reaction amplification using 16S rRNA gene-specific primers and Cy3-dCTP labeling. Labeled DNA was then applied to microbial microarrays (280 species generated by the Human Oral Microbial Identification Microarray Laboratory (Forsyth Institute, Cambridge, MA. Results: The most prevalent microorganisms, found across multiple abscess specimens, include Fusobacterium nucleatum, Parvimonas micra, Megasphaera species clone CS025, Prevotella multisaccharivorax, Atopobium rimae, and Porphyromonas endodontalis. The most abundant microorganisms, found in highest numbers within individual abscesses, include F. nucleatum, P. micra, Streptococcus Cluster III, Solobacterium moorei, Streptococcus constellatus, and Porphyromonas endodontalis. Strong bacterial associations were identified between Prevotella multisaccharivorax, Acidaminococcaceae species clone DM071, Megasphaera species clone CS025, Actinomyces species clone EP053, and Streptococcus cristatus (all with Spearman coefficients >0.9. Conclusions: Cultivable and uncultivable bacterial species have been identified in endodontic abscesses obtained from the Portland, Oregon demographic region, and taxa identifications correlated well with other published studies, with the exception of Treponema and Streptococcus cristae, which

  5. Oral microbiota species in acute apical endodontic abscesses.

    Science.gov (United States)

    George, Noelle; Flamiatos, Erin; Kawasaki, Kellie; Kim, Namgu; Carriere, Charles; Phan, Brian; Joseph, Raphael; Strauss, Shay; Kohli, Richie; Choi, Dongseok; Baumgartner, J Craig; Sedgley, Christine; Maier, Tom; Machida, Curtis A

    2016-01-01

    Acute apical abscesses are serious endodontic diseases resulting from pulpal infection with opportunistic oral microorganisms. The objective of this study was to identify and compare the oral microbiota in patients (N=18) exhibiting acute apical abscesses, originating from the demographic region in Portland, Oregon. The study hypothesis is that abscesses obtained from this demographic region may contain unique microorganisms not identified in specimens from other regions. Endodontic abscesses were sampled from patients at the Oregon Health & Science University (OHSU) School of Dentistry. DNA from abscess specimens was subjected to polymerase chain reaction amplification using 16S rRNA gene-specific primers and Cy3-dCTP labeling. Labeled DNA was then applied to microbial microarrays (280 species) generated by the Human Oral Microbial Identification Microarray Laboratory (Forsyth Institute, Cambridge, MA). The most prevalent microorganisms, found across multiple abscess specimens, include Fusobacterium nucleatum, Parvimonas micra, Megasphaera species clone CS025, Prevotella multisaccharivorax, Atopobium rimae, and Porphyromonas endodontalis. The most abundant microorganisms, found in highest numbers within individual abscesses, include F. nucleatum, P. micra, Streptococcus Cluster III, Solobacterium moorei, Streptococcus constellatus, and Porphyromonas endodontalis. Strong bacterial associations were identified between Prevotella multisaccharivorax, Acidaminococcaceae species clone DM071, Megasphaera species clone CS025, Actinomyces species clone EP053, and Streptococcus cristatus (all with Spearman coefficients >0.9). Cultivable and uncultivable bacterial species have been identified in endodontic abscesses obtained from the Portland, Oregon demographic region, and taxa identifications correlated well with other published studies, with the exception of Treponema and Streptococcus cristae, which were not commonly identified in endodontic abscesses between the

  6. Impact of Periodontal Therapy on the Subgingival Microbiota of Severe Periodontitis: Comparison between Good Responders and “Refractory” Subjects by the Human Oral Microbe Identification Microarray (HOMIM)

    Science.gov (United States)

    Colombo, Ana Paula V.; Bennet, Susan; Cotton, Sean L.; Goodson, J. Max; Kent, Ralph; Haffajee, Anne D.; Socransky, Sigmund S.; Hasturk, Hatice; Van Dyke, Thomas E.; Dewhirst, Floyd E.; Paster, Bruce J.

    2014-01-01

    Aim This study compared the changes on the subgingival microbiota of subjects with “refractory” periodontitis (RP) or treatable periodontitis (GR) before and after periodontal therapy by using the Human Oral Microbe Identification Microarray (HOMIM). Methods Individuals with chronic periodontitis were classified as RP (n=17) based on mean attachment loss (AL) and/or >3 sites with AL ≥2.5 mm after scaling and root planing, surgery and systemically administered amoxicillin and metronidazole or as GR (n=30) based on mean attachment gain and no sites with AL ≥2.5 mm after treatment. Subgingival plaque samples were taken at baseline and 15 months after treatment and analyzed for the presence of 300 species by HOMIM analysis. Significant differences in taxa before and after therapy were sought using the Wilcoxon test. Results The majority of species evaluated decreased in prevalence in both groups after treatment; however, only a small subset of organisms was significantly affected. Species that increased or persisted in high frequency in RP but were significantly reduced in GR included Bacteroidetes sp., Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella spp., Tannerella forsythia, Dialister spp., Selenomonas spp., Catonella morbi, Eubacterium spp., Filifactor alocis, Parvimonas micra, Peptostreptococcus sp. OT113, Fusobacterium sp. OT203, Pseudoramibacter alactolyticus, Streptococcus intermedius or Streptococcus constellatus and Shuttlesworthia satelles. In contrast, Capnocytophaga sputigena, Cardiobacterium hominis, Gemella haemolysans, Haemophilus parainfluenzae, Kingella oralis, Lautropia mirabilis, Neisseria elongata, Rothia dentocariosa, Streptococcus australis and Veillonella spp. were more associated with therapeutic success. Conclusion Persistence of putative and novel periodontal pathogens, as well as low prevalence of beneficial species was associated with chronic “refractory” periodontitis. PMID:22324467

  7. Molecular analysis of 16S rRNA genes identifies potentially periodontal pathogenic bacteria and archaea in the plaque of partially erupted third molars.

    Science.gov (United States)

    Mansfield, J M; Campbell, J H; Bhandari, A R; Jesionowski, A M; Vickerman, M M

    2012-07-01

    Small subunit rRNA sequencing and phylogenetic analysis were used to identify cultivable and uncultivable microorganisms present in the dental plaque of symptomatic and asymptomatic partially erupted third molars to determine the prevalence of putative periodontal pathogens in pericoronal sites. Template DNA prepared from subgingival plaque collected from partially erupted symptomatic and asymptomatic mandibular third molars and healthy incisors was used in polymerase chain reaction with broad-range oligonucleotide primers to amplify 16S rRNA bacterial and archaeal genes. Amplicons were cloned, sequenced, and compared with known nucleotide sequences in online databases to identify the microorganisms present. Two thousand three hundred two clones from the plaque of 12 patients carried bacterial sequences from 63 genera belonging to 11 phyla, including members of the uncultivable TM7, SR1, and Chloroflexi, and difficult-to-cultivate Synergistetes and Spirochaetes. Dialister invisus, Filifactor alocis, Fusobacterium nucleatum, Porphyromonas endodontalis, Prevotella denticola, Tannerella forsythia, and Treponema denticola, which have been associated with periodontal disease, were found in significantly greater abundance in pericoronal compared with incisor sites. Dialister invisus and F nucleatum were found in greater abundance in sites exhibiting clinical symptoms. The archaeal species, Methanobrevibacter oralis, which has been associated with severe periodontitis, was found in 3 symptomatic patients. These findings have provided new insights into the complex microbiota of pericoronitis. Several bacterial and archaeal species implicated in periodontal disease were recovered in greater incidence and abundance from the plaque of partially erupted third molars compared with incisors, supporting the hypothesis that the pericoronal region may provide a favored niche for periodontal pathogens in otherwise healthy mouths. Copyright © 2012 American Association of Oral and

  8. Co-localized or randomly distributed? Pair cross correlation of in vivo grown subgingival biofilm bacteria quantified by digital image analysis.

    Directory of Open Access Journals (Sweden)

    Claudia Schillinger

    Full Text Available The polymicrobial nature of periodontal diseases is reflected by the diversity of phylotypes detected in subgingival plaque and the finding that consortia of suspected pathogens rather than single species are associated with disease development. A number of these microorganisms have been demonstrated in vitro to interact and enhance biofilm integration, survival or even pathogenic features. To examine the in vivo relevance of these proposed interactions, we extended the spatial arrangement analysis tool of the software daime (digital image analysis in microbial ecology. This modification enabled the quantitative analysis of microbial co-localization in images of subgingival biofilm species, where the biomass was confined to fractions of the whole-image area, a situation common for medical samples. Selected representatives of the disease-associated red and orange complexes that were previously suggested to interact with each other in vitro (Tannerella forsythia with Fusobacterium nucleatum and Porphyromonas gingivalis with Prevotella intermedia were chosen for analysis and labeled with specific fluorescent probes via fluorescence in situ hybridization. Pair cross-correlation analysis of in vivo grown biofilms revealed tight clustering of F. nucleatum/periodonticum and T. forsythia at short distances (up to 6 µm with a pronounced peak at 1.5 µm. While these results confirmed previous in vitro observations for F. nucleatum and T. forsythia, random spatial distribution was detected between P. gingivalis and P. intermedia in the in vivo samples. In conclusion, we successfully employed spatial arrangement analysis on the single cell level in clinically relevant medical samples and demonstrated the utility of this approach for the in vivo validation of in vitro observations by analyzing statistically relevant numbers of different patients. More importantly, the culture-independent nature of this approach enables similar quantitative analyses for "as

  9. The role of IL-1 gene polymorphisms (IL1A, IL1B, and IL1RN) as a risk factor in unsuccessful implants retaining overdentures.

    Science.gov (United States)

    Sampaio Fernandes, Margarida; Vaz, Paula; Braga, Ana Cristina; Sampaio Fernandes, João Carlos; Figueiral, Maria Helena

    2017-10-01

    Implant-supported overdentures are an alternative predictable rehabilitation method that has a high impact on improving the patient's quality of life. However, some biological complications may interfere with the maintenance and survival of these overdenture implants. The goal of this article was to assess the factors that affect peri-implant success, through a hypothetical prediction model for biological complications of implant overdentures. A retrospective observational, prevalence study was conducted in 58 edentulous Caucasian patients rehabilitated with implant overdentures. A total of 229 implants were included in the study. Anamnestic, clinical, and implant-related parameters were collected and recorded in a single database. "Patient" was chosen as the unit of analysis, and a complete screening protocol was established. The data analytical study included assessing the odds ratio, concerning the presence or absence of a particular risk factor, by using binary logistic regression modeling. Probability values (p values) inferior to 0.05 were considered as representing statistically significant evidence. The performed prediction model included the following variables: mean probing depth, metal exposure, IL1B_allele2, maxillary edentulousness, and Fusobacterium nucleatum. The F. nucleatum showed significant association with the outcome. Introducing a negative coefficient appeared to prevent complications or even boost the biological defense when associated with other factors. The prediction model developed in this study could serve as a basis for further improved models that would assist clinicians in the daily diagnosis and treatment planning practice of oral rehabilitation with implant overdentures. Copyright © 2017 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.

  10. Transcriptional responses of Treponema denticola to other oral bacterial species.

    Directory of Open Access Journals (Sweden)

    Juni Sarkar

    Full Text Available The classic organization by Socransky and coworkers categorized the oral bacteria of the subgingival plaque into different complexes. Treponema denticola, Porphyromonas gingivalis and Tannerella forsythia are grouped into the red complex that is highly correlated with periodontal disease. Socransky's work closely associates red with orange complex species such as Fusobacterium nucleatum and Prevotella intermedia but not with members of the other complexes. While the relationship between species contained by these complexes is in part supported by their ability to physically attach to each other, the physiological consequences of these interactions and associations are less clear. In this study, we employed T. denticola as a model organism to analyze contact-dependent responses to interactions with species belonging to the same complex (P. gingivalis and T. forsythia, the closely associated orange complex (using F. nucleatum and P. intermedia as representatives and the unconnected yellow complex (using Streptococcus sanguinis and S. gordonii as representatives. RNA was extracted from T. denticola alone as well as after pairwise co-incubation for 5 hrs with representatives of the different complexes, and the respective gene expression profiles were determined using microarrays. Numerous genes related to motility, metabolism, transport, outer membrane and hypothetical proteins were differentially regulated in T. denticola in the presence of the tested partner species. Further analysis revealed a significant overlap in the affected genes and we identified a general response to the presence of other species, those specific to two of the three complexes as well as individual complexes. Most interestingly, many predicted major antigens (e.g. flagella, Msp, CTLP were suppressed in responses that included red complex species indicating that the presence of the most closely associated species induces immune-evasive strategies. In summary, the data

  11. Microbial profile on metallic and ceramic bracket materials.

    Science.gov (United States)

    Anhoury, Patrick; Nathanson, Dan; Hughes, Christopher V; Socransky, Sigmund; Feres, Magda; Chou, Laisheng Lee

    2002-08-01

    The placement of orthodontic appliances creates a favorable environment for the accumulation of a microbiota and food residues, which, in time, may cause caries or exacerbate any pre-existing periodontal disease. The purpose of the present study was to compare the total bacterial counts present on metallic and ceramic orthodontic brackets in order to clarify which bracket type has a higher plaque retaining capacity and to determine the levels of Streptococcus mutans and Lactobacillus spp on both types of brackets. Thirty-two metallic brackets and 24 ceramic brackets were collected from orthodontic patients at the day of debonding. Two brackets were collected from each patient; one from a maxillary central incisor and another from a maxillary second premolar. Sixteen patients who used metallic brackets and 12 patients who used ceramic brackets were sampled. Bacterial populations were studied using "checkerboard" DNA-DNA hybridization, which uses DNA probes to identify species in complex microbial samples. The significance of differences between groups was determined using the Mann-Whitney U-test. Results showed no significant differences between metallic and ceramic brackets with respect to the caries-inducing S mutans and L acidophilus spp counts. Mean counts of 8 of 35 additional species differed significantly between metallic and ceramic brackets with no obvious pattern favoring one bracket type over the other. This study showed higher mean counts of Treponema denticola, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum ss vincentii, Streptococcus anginosus, and Eubacterium nodatum on metallic brackets while higher counts of Eikenella corrodens, Campylobacter showae, and Selenomonas noxia were found on ceramic brackets.

  12. [Anaerobic bacteria isolated from patients with suspected anaerobic infections].

    Science.gov (United States)

    Ercis, Serpil; Tunçkanat, Ferda; Hasçelik, Gülşen

    2005-10-01

    The study involved 394 clinical samples sent to the Clinical Microbiology Laboratory of Hacettepe University Adult Hospital between January 1997 and May 2004 for anaerobic cultivation. Since multiple cultures from the same clinical samples of the same patient were excluded, the study was carried on 367 samples. The anaerobic cultures were performed in anaerobic jar using AnaeroGen kits (Oxoid, Basingstoke, U.K.) or GENbox (bioMérieux, Lyon, France). The isolates were identified by both classical methods and "BBL Crystal System" (Becton Dickinson, U.S.A.). While no growth was detected in 120 (32.7%) of the clinical samples studied, in 144 samples (39.2%) only aerobes, in 28 (7.6%) only anaerobes and in 75 (20.5%) of the samples both aerobes and anaerobes were isolated. The number of the anaerobic isolates was 217 from 103 samples with anaerobic growth. Of these 103 samples 15 showed single bacterial growth whereas in 88 samples multiple bacterial isolates were detected. Anaerobic isolates consisted of 92 Gram negative bacilli (Bacteroides spp. 50, Prevotella spp. 14, Porphyromonas spp. 10, Fusobacterium spp. 7, Tisierella spp. 2, unidentified 9), 57 Gram positive bacilli (Clostridium spp.17, Propionibacterium spp. 16, Lactobacillus spp. 8, Actinomyces spp. 5, Eubacterium spp. 2, Bifidobacterium adolescentis 1, Mobiluncus mulieris 1, unidentified nonspore forming rods 7), 61 Gram positive cocci (anaerobic cocci 44, microaerophilic cocci 17), and 7 Gram negative cocci (Veillonella spp.). In conclusion, in the samples studied with prediagnosis of anaerobic infection, Bacteroides spp. (23%) were the most common bacteria followed by anaerobic Gram positive cocci (20.3%) and Clostridium spp (7.8%).

  13. Photodynamic therapy as an adjunct to non-surgical periodontal treatment: a randomized, controlled clinical trial.

    Science.gov (United States)

    Christodoulides, Nicos; Nikolidakis, Dimitris; Chondros, Panagiotis; Becker, Jürgen; Schwarz, Frank; Rössler, Ralf; Sculean, Anton

    2008-09-01

    Recent preclinical and clinical data have suggested a potential benefit of photodynamic therapy (PDT) in the treatment of periodontitis. However, there are very limited data from controlled clinical trials evaluating the effect of PDT in the treatment of periodontitis. The aim of this study was to evaluate the clinical and microbiologic effects of the adjunctive use of PDT to non-surgical periodontal treatment. Twenty-four subjects with chronic periodontitis were randomly treated with scaling and root planing followed by a single episode of PDT (test) or scaling and root planing alone (control). Full-mouth plaque score (FMPS), full-mouth bleeding score (FMBS), probing depth (PD), gingival recession, and clinical attachment level (CAL) were measured at baseline and 3 and 6 months after therapy. Primary outcome variables were changes in PD and CAL. Microbiologic evaluation of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia (previously T. forsythensis), Treponema denticola, Parvimonas micra (previously Peptostreptococcus micros or Micromonas micros), Fusobacterium nucleatum, Campylobacter rectus, Eubacterium nodatum, Eikenella corrodens, and Capnocytophaga spp. was performed at baseline and 3 and 6 months following therapy by using a commercially available polymerase chain reaction test. At 3 and 6 months after treatment, there were no statistically significant differences between the groups with regard to CAL, PD, FMPS, or microbiologic changes. At 3 and 6 months, a statistically significantly greater improvement in FMBS was found in the test group. The additional application of a single episode of PDT to scaling and root planing failed to result in an additional improvement in terms of PD reduction and CAL gain, but it resulted in a significantly higher reduction in bleeding scores compared to scaling and root planing alone.

  14. [Periodontal microbiota and microorganisms isolated from heart valves in patients undergoing valve replacement surgery in a clinic in Cali, Colombia].

    Science.gov (United States)

    Moreno, Sandra; Parra, Beatriz; Botero, Javier E; Moreno, Freddy; Vásquez, Daniel; Fernández, Hugo; Alba, Sandra; Gallego, Sara; Castillo, Gilberto; Contreras, Adolfo

    2017-12-01

    Periodontitis is an infectious disease that affects the support tissue of the teeth and it is associated with different systemic diseases, including cardiovascular disease. Microbiological studies facilitate the detection of microorganisms from subgingival and cardiovascular samples. To describe the cultivable periodontal microbiota and the presence of microorganisms in heart valves from patients undergoing valve replacement surgery in a clinic in Cali. We analyzed 30 subgingival and valvular tissue samples by means of two-phase culture medium, supplemented blood agar and trypticase soy agar with antibiotics. Conventional PCR was performed on samples of valve tissue. The periodontal pathogens isolated from periodontal pockets were: Fusobacterium nucleatum (50%), Prevotella intermedia/ nigrescens (40%), Campylobacter rectus (40%), Eikenella corrodens (36.7%), Gram negative enteric bacilli (36.7%), Porphyromonas gingivalis (33.3%), and Eubacterium spp. (33.3%). The pathogens isolated from the aortic valve were Propionibacterium acnes (12%), Gram negative enteric bacilli (8%), Bacteroides merdae (4%), and Clostridium bifermentans (4%), and from the mitral valve we isolated P. acnes and Clostridium beijerinckii. Conventional PCR did not return positive results for oral pathogens and bacterial DNA was detected only in two samples. Periodontal microbiota of patients undergoing surgery for heart valve replacement consisted of species of Gram-negative bacteria that have been associated with infections in extraoral tissues. However, there is no evidence of the presence of periodontal pathogens in valve tissue, because even though there were valve and subgingival samples positive for Gram-negative enteric bacilli, it is not possible to maintain they corresponded to the same phylogenetic origin.

  15. In vitro utilization of amylopectin and high-amylose maize (Amylomaize) starch granules by human colonic bacteria.

    Science.gov (United States)

    Wang, X; Conway, P L; Brown, I L; Evans, A J

    1999-11-01

    It has been well established that a certain amount of ingested starch can escape digestion in the human small intestine and consequently enters the large intestine, where it may serve as a carbon source for bacterial fermentation. Thirty-eight types of human colonic bacteria were screened for their capacity to utilize soluble starch, gelatinized amylopectin maize starch, and high-amylose maize starch granules by measuring the clear zones on starch agar plates. The six cultures which produced clear zones on amylopectin maize starch- containing plates were selected for further studies for utilization of amylopectin maize starch and high-amylose maize starch granules A (amylose; Sigma) and B (Culture Pro 958N). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to detect bacterial starch-degrading enzymes. It was demonstrated that Bifidobacterium spp., Bacteroides spp., Fusobacterium spp., and strains of Eubacterium, Clostridium, Streptococcus, and Propionibacterium could hydrolyze the gelatinized amylopectin maize starch, while only Bifidobacterium spp. and Clostridium butyricum could efficiently utilize high-amylose maize starch granules. In fact, C. butyricum and Bifidobacterium spp. had higher specific growth rates in the autoclaved medium containing high-amylose maize starch granules and hydrolyzed 80 and 40% of the amylose, respectively. Starch-degrading enzymes were cell bound on Bifidobacterium and Bacteroides cells and were extracellular for C. butyricum. Active staining for starch-degrading enzymes on SDS-PAGE gels showed that the Bifidobacterium cells produced several starch-degrading enzymes with high relative molecular (M(r)) weights (>160,000), medium-sized relative molecular weights (>66,000), and low relative molecular weights (spp. and C. butyricum degraded and utilized granules of amylomaize starch.

  16. Impact of periodontal therapy on the subgingival microbiota of severe periodontitis: comparison between good responders and individuals with refractory periodontitis using the human oral microbe identification microarray.

    Science.gov (United States)

    Colombo, Ana Paula V; Bennet, Susan; Cotton, Sean L; Goodson, J Max; Kent, Ralph; Haffajee, Anne D; Socransky, Sigmund S; Hasturk, Hatice; Van Dyke, Thomas E; Dewhirst, Floyd E; Paster, Bruce J

    2012-10-01

    This study compares the changes to the subgingival microbiota of individuals with "refractory" periodontitis (RP) or treatable periodontitis (good responders [GR]) before and after periodontal therapy by using the Human Oral Microbe Identification Microarray (HOMIM) analysis. Individuals with chronic periodontitis were classified as RP (n = 17) based on mean attachment loss (AL) and/or >3 sites with AL ≥2.5 mm after scaling and root planing, surgery, and systemically administered amoxicillin and metronidazole or as GR (n = 30) based on mean attachment gain and no sites with AL ≥2.5 mm after treatment. Subgingival plaque samples were taken at baseline and 15 months after treatment and analyzed for the presence of 300 species by HOMIM analysis. Significant differences in taxa before and post-therapy were sought using the Wilcoxon test. The majority of species evaluated decreased in prevalence in both groups after treatment; however, only a small subset of organisms was significantly affected. Species that increased or persisted in high frequency in RP but were significantly reduced in GR included Bacteroidetes sp., Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella spp., Tannerella forsythia, Dialister spp., Selenomonas spp., Catonella morbi, Eubacterium spp., Filifactor alocis, Parvimonas micra, Peptostreptococcus sp. OT113, Fusobacterium sp. OT203, Pseudoramibacter alactolyticus, Streptococcus intermedius or Streptococcus constellatus, and Shuttlesworthia satelles. In contrast, Capnocytophaga sputigena, Cardiobacterium hominis, Gemella haemolysans, Haemophilus parainfluenzae, Kingella oralis, Lautropia mirabilis, Neisseria elongata, Rothia dentocariosa, Streptococcus australis, and Veillonella spp. were more associated with therapeutic success. Persistence of putative and novel periodontal pathogens, as well as low prevalence of beneficial species was associated with chronic refractory periodontitis.

  17. Antimicrobial susceptibility of clinical isolates of anaerobic bacteria in Ontario, 2010-2011.

    Science.gov (United States)

    Marchand-Austin, Alex; Rawte, Prasad; Toye, Baldwin; Jamieson, Frances B; Farrell, David J; Patel, Samir N

    2014-08-01

    The local epidemiology of antimicrobial susceptibility patterns in anaerobic bacteria is important in guiding the empiric treatment of infections. However, susceptibility data are very limited on anaerobic organisms, particularly among non-Bacteroides organisms. To determine susceptibility profiles of clinically-significant anaerobic bacteria in Ontario Canada, anaerobic isolates from sterile sites submitted to Public Health Ontario Laboratory (PHOL) for identification and susceptibility testing were included in this study. Using the E-test method, isolates were tested for various antimicrobials including, penicillin, cefoxitin, clindamycin, meropenem, piperacillin-tazobactam and metronidazole. The MIC results were interpreted based on guidelines published by Clinical and Laboratory Standards Institute. Of 2527 anaerobic isolates submitted to PHOL, 1412 were either from sterile sites or bronchial lavage, and underwent susceptibility testing. Among Bacteroides fragilis, 98.2%, 24.7%, 1.6%, and 1.2% were resistant to penicillin, clindamycin, piperacillin-tazobactam, and metronidazole, respectively. Clostridium perfringens was universally susceptible to penicillin, piperacillin-tazobactam, and meropenem, whereas 14.2% of other Clostridium spp. were resistant to penicillin. Among Gram-positive anaerobes, Actinomyces spp., Parvimonas micra and Propionibacterium spp. were universally susceptible to β-lactams. Eggerthella spp., Collinsella spp., and Eubacterium spp. showed variable resistance to penicillin. Among Gram-negative anaerobes, Fusobacterium spp., Prevotella spp., and Veillonella spp. showed high resistance to penicillin but were universally susceptible to meropenem and piperacillin-tazobactam. The detection of metronidazole resistant B. fragilis is concerning as occurrence of these isolates is extremely rare. These data highlight the importance of ongoing surveillance to provide clinically relevant information to clinicians for empiric management of

  18. Current Diagnosis and Treatment of Halitosis

    Directory of Open Access Journals (Sweden)

    Mehmet Mustafa Kılıçkaya

    2015-11-01

    Full Text Available Halitosis or oral malodor is not a diagnosis, but is symptom. Halitosis, that we frequently encounter in ear, nose and throat practice can be the harbinger of some serious underlying disease. Therefore, diagnosis and to find the cause of the halitosis are important. Also halitosis treatment is necessary due to the social and psychological effects. Breath contains hundreds of volatile organic compounds that are by-products of our metabolism. Certain diseases such as nasopharynx cancer, larynx cancer ve lung cancer alter the mix of gases. Thus, the analysis of exhaled air has gained importance. New technologies lead to the development of new devices. And with these called electronic noses the analysis of exhaled air has becomes an important non-invasive diagnostic method. In the literature, halitosis and bad breath which is used as synonymus with oral malodor is the emission of unpleasant odor from mouth and nasal passage. It occurs in 25% of the population, approximately and it has a significant social and economic impact. Halitosis is classified as true halitosis (physiologic halitosis and pathologic halitosis, pseudohalitosis and halitophobia. The most common cause is intra-oral diseases. Among all these factors, the most important etiologic factor are the coating tongue. Other ear, nose and throat diseases such as rhinitis and sinusitis are seen among the most common extraoral causes. Treponema denticola, Porphyromonas gingivalis, Tanneralla forsythia, Fusobacterium nucleatum, Prevotella intermedia, Prevotella nigrescens, Actinobacilli and Solobacterium moorei are the bacteria which are commonly isolated from patients with halitosis and they are volatile sulfur compounds (VSCs producing ones as well. The treatment of halitosis should be carried out according to the etiology. In the physiologic halitosis tooth brushing, use of dental floss, tongue cleaning and chlorhexidine, cetylpyridinium chloride and zinc containing antimicrobial mouthwashes

  19. Effect of non-surgical periodontal treatment on the subgingival microbiota of patients with chronic kidney disease

    Directory of Open Access Journals (Sweden)

    Hilana Paula Carillo Artese

    2012-08-01

    Full Text Available This study investigated the effect of non-surgical periodontal therapy on the composition of subgingival microbiota of patients with chronic kidney disease (CKD. Sixteen CKD pre-dialysis individuals (CKD and 14 individuals without clinical evidence of kidney disease (C presenting chronic periodontitis were treated by scaling and root planing. Subgingival samples were collected from each patient and analyzed for their composition by checkerboard at baseline and 3 months post-therapy. Significant differences between groups at baseline were sought by the Mann-Whitney and χ² tests. Changes over time were examined by the Wilcoxon test. At baseline, the CKD group had significantly lower counts of E. faecalis compared to the C group (p < 0.05. After treatment, the levels of a greater number of species were reduced in the C group. Higher levels of A. israelii, C. rectus, F. periodonticum, P. micra, P. nigrescens, T. forsythia, N. mucosa, and S. anginosus (p < 0.05 were found in the CKD group compared to the C group. Also, non-responsive sites in CKD individuals harbored significantly higher levels of pathogenic species (T. forsythia, P. gingivalis, T. denticola, Fusobacterium spp., D. pneumosintes, E. faecalis and S. aureus; p < 0.05 than sites that responded to therapy, as well as non-responsive sites in the C group. The periodontitis-associated subgingival microbiota of CKD and systemically healthy individuals was similar in composition. However, high levels of pathogenic species persisted in the subgingival microbiota of patients with CKD after treatment.

  20. Intra-Genomic Heterogeneity in 16S rRNA Genes in Strictly Anaerobic Clinical Isolates from Periodontal Abscesses.

    Science.gov (United States)

    Chen, Jiazhen; Miao, Xinyu; Xu, Meng; He, Junlin; Xie, Yi; Wu, Xingwen; Chen, Gang; Yu, Liying; Zhang, Wenhong

    2015-01-01

    Members of the genera Prevotella, Veillonella and Fusobacterium are the predominant culturable obligate anaerobic bacteria isolated from periodontal abscesses. When determining the cumulative number of clinical anaerobic isolates from periodontal abscesses, ambiguous or overlapping signals were frequently encountered in 16S rRNA gene sequencing chromatograms, resulting in ambiguous identifications. With the exception of the genus Veillonella, the high intra-chromosomal heterogeneity of rrs genes has not been reported. The 16S rRNA genes of 138 clinical, strictly anaerobic isolates and one reference strain were directly sequenced, and the chromatograms were carefully examined. Gene cloning was performed for 22 typical isolates with doublet sequencing signals for the 16S rRNA genes, and four copies of the rrs-ITS genes of 9 Prevotella intermedia isolates were separately amplified by PCR, sequenced and compared. Five conserved housekeeping genes, hsp60, recA, dnaJ, gyrB1 and rpoB from 89 clinical isolates of Prevotella were also amplified by PCR and sequenced for identification and phylogenetic analysis along with 18 Prevotella reference strains. Heterogeneity of 16S rRNA genes was apparent in clinical, strictly anaerobic oral bacteria, particularly in the genera Prevotella and Veillonella. One hundred out of 138 anaerobic strains (72%) had intragenomic nucleotide polymorphisms (SNPs) in multiple locations, and 13 strains (9.4%) had intragenomic insertions or deletions in the 16S rRNA gene. In the genera Prevotella and Veillonella, 75% (67/89) and 100% (19/19) of the strains had SNPs in the 16S rRNA gene, respectively. Gene cloning and separate amplifications of four copies of the rrs-ITS genes confirmed that 2 to 4 heterogeneous 16S rRNA copies existed. Sequence alignment of five housekeeping genes revealed that intra-species nucleotide similarities were very high in the genera Prevotella, ranging from 94.3-100%. However, the inter-species similarities were

  1. Preliminary characterization of the oral microbiota of Chinese adults with and without gingivitis

    Directory of Open Access Journals (Sweden)

    Huang Shi

    2011-12-01

    Full Text Available Abstract Background Microbial communities inhabiting human mouth are associated with oral health and disease. Previous studies have indicated the general prevalence of adult gingivitis in China to be high. The aim of this study was to characterize in depth the oral microbiota of Chinese adults with or without gingivitis, by defining the microbial phylogenetic diversity and community-structure using highly paralleled pyrosequencing. Methods Six non-smoking Chinese, three with and three without gingivitis (age range 21-39 years, 4 females and 2 males were enrolled in the present cross-sectional study. Gingival parameters of inflammation and bleeding on probing were characterized by a clinician using the Mazza Gingival Index (MGI. Plaque (sampled separately from four different oral sites and salivary samples were obtained from each subject. Sequences and relative abundance of the bacterial 16 S rDNA PCR-amplicons were determined via pyrosequencing that produced 400 bp-long reads. The sequence data were analyzed via a computational pipeline customized for human oral microbiome analyses. Furthermore, the relative abundances of selected microbial groups were validated using quantitative PCR. Results The oral microbiomes from gingivitis and healthy subjects could be distinguished based on the distinct community structures of plaque microbiomes, but not the salivary microbiomes. Contributions of community members to community structure divergence were statistically accessed at the phylum, genus and species-like levels. Eight predominant taxa were found associated with gingivitis: TM7, Leptotrichia, Selenomonas, Streptococcus, Veillonella, Prevotella, Lautropia, and Haemophilus. Furthermore, 98 species-level OTUs were identified to be gingivitis-associated, which provided microbial features of gingivitis at a species resolution. Finally, for the two selected genera Streptococcus and Fusobacterium, Real-Time PCR based quantification of relative bacterial

  2. Nasopharyngeal Microbiome Diversity Changes over Time in Children with Asthma.

    Science.gov (United States)

    Pérez-Losada, Marcos; Alamri, Lamia; Crandall, Keith A; Freishtat, Robert J

    2017-01-01

    The nasopharynx is a reservoir for pathogens associated with respiratory illnesses such as asthma. Next-generation sequencing (NGS) has been used to characterize the nasopharyngeal microbiome of infants and adults during health and disease; less is known, however, about the composition and temporal dynamics (i.e., longitudinal variation) of microbiotas from children and adolescents. Here we use NGS technology to characterize the nasopharyngeal microbiomes of asthmatic children and adolescents (6 to 18 years) and determine their stability over time. Two nasopharyngeal washes collected 5.5 to 6.5 months apart were taken from 40 children and adolescents with asthma living in the Washington D.C. area. Sequence data from the 16S-V4 rRNA gene region (~250 bp) were collected from the samples using the MiSeq platform. Raw data were processed in mothur (SILVA123 reference database) and Operational Taxonomic Units (OTU)-based alpha- and beta-diversity metrics were estimated. Relatedness among samples was assessed using PCoA ordination and Procrustes analyses. Differences in microbial diversity and taxon mean relative proportions were assessed using linear mixed effects models. Core microbiome analyses were also performed to identify stable and consistent microbes of the nasopharynx. A total of 2,096,584 clean 16S sequences corresponding to an average of 167 OTUs per sample were generated. Representatives of Moraxella*, Staphylococcus*, Dolosigranulum, Corynebacterium, Prevotella, Streptococcus*, Haemophilus*, Fusobacterium* and a Neisseriaceae genus accounted for 86% of the total reads. These nine genera have been previously found in the nasopharynxes of both infants and adults, but in different proportions. OTUs from the five genera highlighted (*) above defined the nasopharyngeal core microbiome at the 95% level. No significant differences in alpha- and beta-diversity were observed between seasons, but bacterial mean relative proportions of Haemophilus, Moraxella

  3. Nasopharyngeal Microbiome Diversity Changes over Time in Children with Asthma.

    Directory of Open Access Journals (Sweden)

    Marcos Pérez-Losada

    Full Text Available The nasopharynx is a reservoir for pathogens associated with respiratory illnesses such as asthma. Next-generation sequencing (NGS has been used to characterize the nasopharyngeal microbiome of infants and adults during health and disease; less is known, however, about the composition and temporal dynamics (i.e., longitudinal variation of microbiotas from children and adolescents. Here we use NGS technology to characterize the nasopharyngeal microbiomes of asthmatic children and adolescents (6 to 18 years and determine their stability over time.Two nasopharyngeal washes collected 5.5 to 6.5 months apart were taken from 40 children and adolescents with asthma living in the Washington D.C. area. Sequence data from the 16S-V4 rRNA gene region (~250 bp were collected from the samples using the MiSeq platform. Raw data were processed in mothur (SILVA123 reference database and Operational Taxonomic Units (OTU-based alpha- and beta-diversity metrics were estimated. Relatedness among samples was assessed using PCoA ordination and Procrustes analyses. Differences in microbial diversity and taxon mean relative proportions were assessed using linear mixed effects models. Core microbiome analyses were also performed to identify stable and consistent microbes of the nasopharynx.A total of 2,096,584 clean 16S sequences corresponding to an average of 167 OTUs per sample were generated. Representatives of Moraxella*, Staphylococcus*, Dolosigranulum, Corynebacterium, Prevotella, Streptococcus*, Haemophilus*, Fusobacterium* and a Neisseriaceae genus accounted for 86% of the total reads. These nine genera have been previously found in the nasopharynxes of both infants and adults, but in different proportions. OTUs from the five genera highlighted (* above defined the nasopharyngeal core microbiome at the 95% level. No significant differences in alpha- and beta-diversity were observed between seasons, but bacterial mean relative proportions of Haemophilus

  4. PIRATE project: point-of-care, informatics-based randomised controlled trial for decreasing overuse of antibiotic therapy in Gram-negative bacteraemia.

    Science.gov (United States)

    Huttner, Angela; Albrich, Werner C; Bochud, Pierre-Yves; Gayet-Ageron, Angèle; Rossel, Anne; Dach, Elodie von; Harbarth, Stephan; Kaiser, Laurent

    2017-07-13

    Antibiotic overuse drives antibiotic resistance. The optimal duration of antibiotic therapy for Gram-negative bacteraemia (GNB), a common community and hospital-associated infection, remains unknown and unstudied via randomised controlled trials (RCTs). This investigator-initiated, multicentre, non-inferiority, informatics-based point-of-care RCT will randomly assign adult hospitalised patients receiving microbiologically efficacious antibiotic(s) for GNB to (1) 14 days of antibiotic therapy, (2) 7 days of therapy or (3) an individualised duration determined by clinical response and 75% reduction in peak C reactive protein (CRP) values. The randomisation will occur in equal proportions (1:1:1) on day 5 (±1) of efficacious antibiotic therapy as determined by antibiogram; patients, their physicians and study investigators will be blind to treatment duration allocation until the day of antibiotic discontinuation. Immunosuppressed patients and those with GNB due to complicated infections (endocarditis, osteomyelitis, etc) and/or non-fermenting bacilli ( Acinetobacter spp, Burkholderia spp, Pseudomonas spp) Brucella spp, Fusobacterium spp or polymicrobial growth with Gram-positive organisms will be ineligible. The primary outcome is incidence of clinical failure at day 30; secondary outcomes include clinical failure, all-cause mortality and incidence of Clostridiumdifficile infection in the 90-day study period. An interim safety analysis will be performed after the first 150 patients have been followed for ≤30 days. Given a chosen margin of 10%, the required sample size to determine non-inferiority is roughly 500 patients. Analyses will be performed on both intention-to-treat and per-protocol populations. Ethics approval was obtained from the cantonal ethics committees of all three participating sites. Results of the main trial and each of the secondary endpoints will be submitted for publication in a peer-reviewed journal. This trial is registered at www

  5. The human jejunum has an endogenous microbiota that differs from those in the oral cavity and colon.

    Science.gov (United States)

    Sundin, Olof H; Mendoza-Ladd, Antonio; Zeng, Mingtao; Diaz-Arévalo, Diana; Morales, Elisa; Fagan, B Matthew; Ordoñez, Javier; Velez, Philip; Antony, Nishaal; McCallum, Richard W

    2017-07-17

    The upper half of the human small intestine, known as the jejunum, is the primary site for absorption of nutrient-derived carbohydrates, amino acids, small peptides, and vitamins. In contrast to the colon, which contains 10 11 -10 12 colony forming units of bacteria per ml (CFU/ml), the normal jejunum generally ranges from 10 3 to 10 5  CFU per ml. Because invasive procedures are required to access the jejunum, much less is known about its bacterial microbiota. Bacteria inhabiting the jejunal lumen have been investigated by classical culture techniques, but not by culture-independent metagenomics. The lumen of the upper jejunum was sampled during enteroscopy of 20 research subjects. Culture on aerobic and anaerobic media gave live bacterial counts ranging from 5.8 × 10 3 CFU/ml to 8.0 × 10 6 CFU/ml. DNA from the same samples was analyzed by 16S rRNA gene-specific quantitative PCR, yielding values from 1.5 × 10 5 to 3.1 × 10 7 bacterial genomes per ml. When calculated for each sample, estimated bacterial viability ranged from effectively 100% to a low of 0.3%. 16S rRNA metagenomic analysis of uncultured bacteria by Illumina MiSeq sequencing gave detailed microbial composition by phylum, genus and species. The genera Streptococcus, Prevotella, Veillonella and Fusobacterium, were especially abundant, as well as non-oral genera including Escherichia, Klebsiella, and Citrobacter. The jejunum was devoid of the genera Alistipes, Ruminococcus, Faecalibacterium, and other extreme anaerobes abundant in the colon. In patients with higher bacterial loads, there was no significant change in microbial species composition. The jejunal lumen contains a distinctive bacterial population consisting primarily of facultative anaerobes and oxygen-tolerant obligate anaerobes similar to those found in the oral cavity. However, the frequent abundance of Enterobacteriaceae represents a major difference from oral microbiota. Although a few genera are shared with the colon, we