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Sample records for actinobacillus lignieresii

  1. Multifocal suppurative granuloma caused by Actinobacillus lignieresii in the peritoneum of a beef steer

    Science.gov (United States)

    KASUYA, Kazufumi; MANCHANAYAKE, Tilusha; UENOYAMA, Kei; KAWA, Sayaka; TAKAYAMA, Kou; IMAI, Naoto; SHIBAHARA, Tomoyuki

    2016-01-01

    An imported crossbred Angus beef steer aged eight to twelve months died suddenly on the eighth day of a quarantine period in Japan. Gross examination showed the peritoneum and mesentery consisted of numerous nodules of various sizes. Histological examination revealed chronic suppurative granulomatous peritonitis with eosinophilic rosettes surrounding colonies of Gram-negative bacilli. The bacteria isolated from the nodules were confirmed to be Actinobacillus lignieresii based on the results of 16S rRNA gene sequencing and immunohistochemistry. Antibiotic sensitivity testing showed that the isolate was resistant to penicillin. Thus, a diagnosis of atypical actinobacillosis caused by A. lignieresii was made. PMID:27773882

  2. Genetic diversity of Actinobacillus lignieresii isolates from different hosts

    DEFF Research Database (Denmark)

    Kokotovic, Branko; Angen, Øystein; Bisgaard, Magne

    2011-01-01

    Genetic diversity detected by analysis of amplified fragment length polymorphisms (AFLPs) of 54 Actinobacilus lignieresii isolates from different hosts and geographic localities is described. On the basis of variances in AFLP profiles, the strains were grouped in two major clusters; one comprisin...

  3. Susceptibility to hydrophobic molecules and phospholipid composition in Pasteurella multocida and Actinobacillus lignieresii.

    OpenAIRE

    1988-01-01

    Despite its typically gram-negative cell envelope ultrastructure, Pasteurella multocida is susceptible to the hydrophobic antibiotic novobiocin and is unable to initiate growth on MacConkey agar, a parameter often used to effect is differentiation from other members of the family Pasteurellaceae such as Actinobacillus lignieresii. However, growth on basal medium supplemented with individual selective factors and an agar diffusion assay revealed the bile salts contained in MacConkey agar to be...

  4. Susceptibility to hydrophobic molecules and phospholipid composition in Pasteurella multocida and Actinobacillus lignieresii.

    Science.gov (United States)

    Hart, M E; Champlin, F R

    1988-09-01

    Despite its typically gram-negative cell envelope ultrastructure, Pasteurella multocida is susceptible to the hydrophobic antibiotic novobiocin and is unable to initiate growth on MacConkey agar, a parameter often used to effect is differentiation from other members of the family Pasteurellaceae such as Actinobacillus lignieresii. However, growth on basal medium supplemented with individual selective factors and an agar diffusion assay revealed the bile salts contained in MacConkey agar to be toxic to both organisms. Four P. multocida surface hydrophobicity variants exhibited consistent in vitro susceptibility to the hydrophobic antibiotics novobiocin, rifamycin SV, and actinomycin D as determined by broth dilution. Readily extractable lipid fractions were obtained by chloroform-methanol extraction of freeze-dried whole cells from exponential-phase cultures. No major differences in total cellular readily extractable lipid content were observed among the P. multocida and A. lignieresii strains examined, although hydrophobic P. multocida strains appeared to contain slightly more than did hydrophilic strains. Analytical thin-layer chromatography and quantitation of resolved readily extractable lipid components revealed the major cell envelope phospholipids of both organisms to be phosphatidylethanolamine and phosphatidylglycerol in a molar ratio of approximately 4:1 regardless of cell surface hydrophobicity properties. Similar results were obtained for Pseudomonas aeruginosa, which is notably refractory to hydrophobic molecules. These data support the conclusion that the permeability of the P. multocida cell envelope to structurally unrelated, hydrophobic molecules is not dependent on cell surface hydrophobicity and cannot be explained on the basis of anomalous polar lipid composition.

  5. Final classification of Bisgaard taxon 9 as Actinobacillus arthritidis sp nov and recognition of a novel genomospecies for equine strains of Actinobacillus lignieresii

    DEFF Research Database (Denmark)

    Christensen, Henrik; Bisgaard, Magne; Angen, Øystein;

    2002-01-01

    Phenotypic characterization of bacteria from diseased and healthy horses identified 18 isolates as Bisgaard taxon 9 and 11 isolates as Actinobacillus lignieresii. All strains of taxon 9 were alpha-galactosidase- and raffinose-positive and showed variable fermentation of (+)L-arabinose and (-)D-sorbitol....... Strains of A. lignieresii were negative for these characteristics, with the exception of raffinose. Two strains from the (-)D-sorbitol-negative group of taxon 9 showed a 16S rRNA similarity of 99.6%, while 99.5% similarity was found between two strains of the (-)D-sorbitol-positive group. DNA......-DNA hybridization between the two strains representing the (-)D-sorbitol-negative group showed 98% binding, and their closest relationship was to a strain of A. lignieresii (64%). The two strains of the (-)D-sorbitol-positive group showed 83% binding and were related to the (-)D-sorbitol-negative group at a 76% DNA...

  6. The transferrin receptor of Actinobacillus pleuropneumoniae: Quantitation of expression and structural characterization using a peptide-specific monoclonal antibody

    DEFF Research Database (Denmark)

    Bøg, Yang S.; Andresen, Lars Ole; Bastholm, L.;

    2001-01-01

    antigens were detected with the Mab in iron-starved Actinobacillus lignieresii, Actinobacillus porcinus, Actinobacillus minor Haemophilus influenzae. and Haemophilus parasuis. Using an enzyme-linked immunosorbent assay (ELISA) based on the Mab 1.48, Tbp2 could be detected in both recombinant E. coli...

  7. Differentiation of Actinobacillus pleuropneumoniae strains by sequence analysis of 16S rDNA and ribosomal intergenic regions, and development of a species specific oligonucleotide for in situ detection

    DEFF Research Database (Denmark)

    Fussing, Vivian; Paster, Bruce J.; Dewhirst, Floyd E.;

    1998-01-01

    The aims of this study were to characterize and determine intraspecies and interspecies relatedness of Actinobacillus pleuropneumoniae to Actinobacillus lignieresii and Actinobacillus suis by sequence analysis of the ribosomal operon and to find a species-specific area for in situ detection of A...

  8. Distribution of diaminopropane, putrescine and cadaverine in Haemophilus and Actinobacillus.

    Science.gov (United States)

    Hamana, K; Nakata, K

    2000-01-01

    Cellular levels of diaminopropane, putrescine and cadaverine, and decarboxylase activities to produce these diamines in six species (16 strains) of Haemophilus and four species (5 strains) of Actinobacillus belonging to the family Pasteurellaceae of the gamma subclass of the class Proteobacteria, were determined by high performance liquid chromatography (HPLC). Diaminopropane was ubiquitously distributed within all Haemophilus and Actinobacillus species, and L-2,4-diaminobutyric acid decarboxylase activity was detected in them. Putrescine and ornithine decarboxylase activity were found in H. aphrophilus, H. parainfluenzae and H. influenzae (type a, b, d, e and f except for type c) but not detected in H. aegyptius, H. parahaemolyticus, H. ducreyi and Actinobacillus species. Cadaverine occurred in H. aphrophilus, H. aegyptius, H. influenzae, H. parainfluenzae, A. actinomycetemcomitans, A. equuli and A. lignieresii, whereas their lysine decarboxylase activity was scarcely detected. Cadaverine was not found in H. parahaemolyticus, H. ducreyi and A. suis. The diamine profile serves as a phenotypic marker for the chemotaxonomic classification of the family Pasteurellaceae.

  9. Clinical significance and taxonomy of Actinobacillus hominis

    DEFF Research Database (Denmark)

    Friis-Møller, Alice; Christensen, J J; Fussing, V;

    2001-01-01

    Clinical findings in 36 immunosuppressed patients with lower respiratory tract infection or bacteremia with Actinobacillus hominis are described. Animal contact was only recorded for three patients; nine patients died despite appropriate antimicrobial treatment. Although infections with this micr...

  10. Fluoroquinolones in the treatment of Actinobacillus actinomycetemcomitans associated periodontitis

    NARCIS (Netherlands)

    Kleinfelder, JW; Mueller, RF; Lange, DE

    2000-01-01

    Background: Periodontitis patients harboring Actinobacillus actinmycetemcomitans (Aa) are prime candidates for systemic antibiotic therapy. Besides tetracycline and the combination of metronidazole and amoxicillin the fluoroquinolones are also believed to have antibacterial activity against Aa. The

  11. [Role of Actinobacillus actinomycetemcomitans in human infection].

    Science.gov (United States)

    Giglio, C; Aránguiz, V; Giglio, M S; Fernández, A

    1990-04-01

    Actinobacillus actinomycetemcomitans (AA), is a cocobacillus thin and small, non motile, uncapsulate and capnophilic. AA, is: one of the species encountered in the mouth's comensal flora being able to be isolated in gingival crevices culture and oral mucosa in a 20% of the healthy population. An important number of pathogenic factors make it well equipped, to protect itself from host's defense mechanisms, and to destroy the periodontal tissue. Between the most important we find lipopolisacarides and leucotoxines which promote tisular invasion and destructive qualities of this microorganism. Since 1912, there are numerous reports of infectious process associated to it, between which we find: endocarditis in native and prothesic valve, soft tissues abscess, pneumonia, brain's abscess, urethritis, vertebral osteomielitis, thyroid's abscess, pericarditis and periodontal juvenile illness, being this one in which its isolation is more frequent. In vitro, AA is very susceptible to tetracicline. This antibiotic reaches high concentrations in gingival crevices, has significant affinity to the alveolar bone and contributes to protect the collagen. These special feature make them the election drug in periodontal disease produced by this microorganism.

  12. Genome Sequence of Actinobacillus suis Type Strain ATCC 33415T.

    Science.gov (United States)

    Calcutt, Michael J; Foecking, Mark F; Mhlanga-Mutangadura, Tendai; Reilly, Thomas J

    2014-09-18

    The assembled and annotated genome of Actinobacillus suis ATCC 33415(T) is reported here. The 2,501,598-bp genome encodes 2,246 open reading frames (ORFs) with strain variable incursion of an integrative conjugative element into a tRNA locus. Comparative analysis of the deduced gene set should inform our understanding of pathogenesis, genomic plasticity, and serotype variation.

  13. Isolation of Actinobacillus suis from a cat's lung

    DEFF Research Database (Denmark)

    Daignault, D.; Chouinard, L.; Møller, Kristian

    1999-01-01

    Actinobacillus suis has been isolated from the lungs of a 9-month-old cat. The bacterium was characterized biochemically as well as genetically, and its sensitivity profile to different antimicrobial agents was established. The role of this isolate in the cat's condition is discussed....

  14. Detection of an Actinobacillus pleuropneumoniae serotype 2 lipopolysaccharide (LPS) variant

    DEFF Research Database (Denmark)

    Stenbaek, E.I.; HovindHaugen, K.

    1996-01-01

    Until now 12 serotypes of Actinobacillus pleuropneumoniae have been recognized. The specificity of the serotypes reside in the carbohydrate composition of the capsular polysaccharides and lipopolysaccharides (LPS). The LPS of A. pleuropneumoniae serotype 2 is a smooth type LPS with O-chains of li......Until now 12 serotypes of Actinobacillus pleuropneumoniae have been recognized. The specificity of the serotypes reside in the carbohydrate composition of the capsular polysaccharides and lipopolysaccharides (LPS). The LPS of A. pleuropneumoniae serotype 2 is a smooth type LPS with O......-chains of linear repeating pentasaccharide units with an O-acetyl group linked to a glucose unit. A monoclonal antibody (MAb 102-G02) directed against A. pleuropneumoniae serotype 2 was characterized in enzyme linked immunosorbent assay (ELISA) and in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS...

  15. Oxidative and nonoxidative killing of Actinobacillus actinomycetemcomitans by human neutrophils.

    OpenAIRE

    Miyasaki, K T; Wilson, M E; Brunetti, A J; Genco, R J

    1986-01-01

    Actinobacillus actinomycetemcomitans is a facultative gram-negative microorganism which has been implicated as an etiologic agent in localized juvenile periodontitis and in subacute bacterial endocarditis and abscesses. Although resistant to serum bactericidal action and to oxidant injury mediated by superoxide anion (O2-) and hydrogen peroxide (H2O2), this organism is sensitive to killing by the myeloperoxidase-hydrogen peroxide-chloride system (K.T. Miyasaki, M.E. Wilson, and R.J. Genco, In...

  16. Osteomyelitis of the mandible due to Aggregatibacter (Actinobacillus actinomycetemcomitans

    Directory of Open Access Journals (Sweden)

    Antony Beena

    2009-01-01

    Full Text Available Aggregatibacter (Actinobacillus actinomycetemcomitans is a capnoic gram negative coccobacilli known to produce juvenile periodontitis. This organism was isolated in pure culture from an unusual case of osteomyelitis of the mandible. The patient was treated with tetracycline, which is the drug of choice for A. actinomycetemcomitans and the clinical response improved. From our limited review of the literature, it appears that this is the first case of osteomyelitis due to A.actinomycetemcomitans reported in India.

  17. Actinobacillus equuli subsp. equuli associated with equine valvular endocarditis

    DEFF Research Database (Denmark)

    Aalbæk, Bent; Østergaard, Stine; Buhl, Rikke;

    2007-01-01

    Microbiological and pathological data from a case of equine valvular endocarditis are reported. Limited information is available on the pathogenic potential of equine Actinobacillus species as several strains originate from apparently healthy horses. After the establishment of two subspecies within...... this species, this seems to be the first report of an etiological association between A. equuli subsp. equuli and equine endocarditis. Furthermore, new information on some phenotypical characteristics of this subspecies are reported, compared to previous findings...

  18. Transmisión intrafamiliar del Actinobacillus actinomycetemcomitans

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    A. Navarro

    2000-05-01

    Full Text Available En la literatura científica existen varias líneas de evidencia que relacionan directamente al Actinobacillus actinomycetemcomitans con lesiones de Periodontitis juvenil localizada y aunque existe evidencia de transmisión familiar de este patógeno periodontal, no existe constancia de que la enfermedad periodontal sea contagiosa. Las bacterias responsables de la enfermedad periodontal parecen ser transmisibles, pero sólo después de un periodo largo de exposición. La vía de transmisión tampoco está clara. Por el momento no es posible sacar ninguna conclusión al respecto.In scientific literature several lines of evidence exist and link Actinobacillus actinomycetemcomitans with localized juvenile periodontitis lesions directly. Although evidence of familial transmission exist, it doesn't prove periodontal disease is contagious. Bacteria responsible for periodontal disease seem to be transmissible, but only after a long exposure period. No clear transmission paths were observed in the population yet. Up to now drawing conclusions about it is not possible.

  19. Serological characterization of Actinobacillus pleuropneumoniae biotype 1 strains antigenically related to both serotypes 2 and 7

    DEFF Research Database (Denmark)

    Nielsen, R.; Andresen, Lars Ole; Plambeck, Tamara

    1996-01-01

    Nine Danish Actinobacillus pleuropneumoniae biotype 1 isolates were shown by latex agglutination and indirect haemagglutination to possess capsular polysaccharide epitopes identical to those of serotype 2 strain 1536 (reference strain of serotype 2) and strain 4226 (Danish serotype 2 strain...

  20. Actinomycetemcomitin: a new bacteriocin produced by Aggregatibacter (Actinobacillus) actinomycetemcomitans.

    Science.gov (United States)

    Lima, Francisca Lúcia; de Carvalho, Maria Auxiliadora Roque; Apolônio, Ana Carolina Morais; Bemquerer, Marcelo Porto; Santoro, Marcelo Matos; Oliveira, Jamil Silvano; Alviano, Celuta Sales; Farias, Luiz de Macêdo

    2008-02-01

    Aggregatibacter (Actinobacillus) actinomycetemcomitans P(7-20) strain isolated from a periodontally diseased patient has produced a bacteriocin (named as actinomycetemcomitin) that is active against Peptostreptococcus anaerobius ATCC 27337. Actinomycetemcomitin was produced during exponential and stationary growth phases, and its amount decreased until it disappeared during the decline growth phase. It was purified by ammonium sulphate precipitation (30-60% saturation), and further by FPLC (mono-Q ionic exchange and Phenyl Superose hydrophobic interaction) and HPLC (C-18 reversed-phase). This bacteriocin loses its activity after incubation at a pH below 7.0 or above 8.0, following heating for 30 min at 45 degrees C, and after treatment with proteolytic enzymes such as trypsin, alpha-chymotrypsin, and papain. Actinomycetemcomitin has a molecular mass of 20.3 KDa and it represents a new bacteriocin from A. actinomycetemcomitans.

  1. The antibacterial mechanism of berberine against Actinobacillus pleuropneumoniae.

    Science.gov (United States)

    Kang, Shuai; Li, Zhengwen; Yin, Zhongqiong; Jia, Renyong; Song, Xu; Li, Li; Chen, Zhenzhen; Peng, Lianci; Qu, Jing; Hu, Zhiqiang; Lai, Xin; Wang, Guangxi; Liang, Xiaoxia; He, Changliang; Yin, Lizi

    2015-01-01

    This study demonstrated berberine to be a potential natural compound against Actinobacillus pleuropneumoniae. Liquid doubling dilution, transmission electron microscopy (TEM), SDS-PAGE and 4',6-diamidino-2-phenylindole (DAPI) staining were employed to elucidate the antibacterial activity and mechanism of berberine. The minimal inhibitory concentration of berberine was 0.3125 mg/mL, and time-kill curves showed concentration and time dependence. The TEM micrographs displayed damaged cell wall, concentrated cytoplasm, cytoplasmic content leakage and cell death. SDS-PAGE and DAPI assays revealed that berberine can restrain DNA and protein syntheses. Berberine inhibited the synthesis of proteins associated with the growth and cleavage of bacteria and then blocked the division and development of bacteria. The compound ultimately induced cytoplasm pyknosis and bacterial death.

  2. Isolation of Actinobacillus pleuropneumoniae serotype 2 by immunomagnetic separation

    DEFF Research Database (Denmark)

    Angen, Øystein; Heegaard, Peter M. H.; Lavritsen, D.T.;

    2001-01-01

    In Denmark porcine pleuropneumonia is most frequently caused by Actinobacillus pleuropneumoniae serotype 2 (60%). Isolation of A. pleuropneumoniae from nasal cavities or tonsils from carrier animals is complicated due to the mixed bacterial flora present. An immunomagnetic separation technique (IMS...... washing steps were performed. The IMS was further evaluated using dilutions of A. pleuropneumoniae with added Pasteurella multocida (10(9) CFU/ml). After two washing steps 15% of the A. pleuropneumoniae cells and no P. multocida was reisolated. A detection limit of 10 CFU/ml was found...... the nasal cavity or tonsils by cultivation or PCR 6 weeks later. By using IMS A. pleuropneumoniae serotype 2 could be reisolated from the tonsils of three pigs. The LMS method represents a valuable tool for isolation of A. pleuropneumoniae from tissue samples....

  3. Immunoglobulin G proteolytic activity of Actinobacillus actinomycetemcomitans Atividade proteolítica de Actinobacillus acitnomycetemcomitans sobre imunoglobulina G

    Directory of Open Access Journals (Sweden)

    Fernanda Akemi Nakanishi

    2006-03-01

    Full Text Available Actinobacillus actinomycetemcomitans produces a protease to human immunoglobulin G that is an important evasion mechanism. In this study, the proteolytic activity of A. actinomycetemcomitans strain ATCC 43718 on human immunoglobulin G associated with culture supernatant concentrations, the growth period and the period of incubation with immunoglobulin G were evaluated by an enzyme linked immunosorbent assay. The protease fraction was detected by Sephadex G 150 chromatography. The results showed that A. actinomycetemcomitans produced a protease to human immunoglobulin G in the culture supernatant, and the highest activity was achieved witen the concentration was 27.5 mug protein/mL, after culturing for 72 hours and incubating with IgG for 24 hours. The molecular mass of the protease active fraction was from 43 to 150 kDa.Actinobacillus actinomycetemcomitans produz protease ativa sobre imunoglobulina G humana, sendo um dos mecanismos importantes de escape do microrganismo. No presente trabalho, foi analisada a atividade proteolítica de sobrenadante de cultivo de A. actinomycetemcomitans ATCC 43718 sobre imunoglobulina G humana em função de concentração, tempo de cultivo do microrganismo e tempo de incubação com IgG, por ensaio imunoenzimático. Adicionalmente, foi determinada a fração com atividade de protease por meio de análise de eluatos de cromatografia em coluna de Sephadex G 150. Os resultados obtidos demonstraram que A. actinomycetemcomitans liberou protease ativa sobre imunoglobulina G humana em sobrenadante de cultivo, sendo a sua maior atividade evidenciada na concentração de 27,5 mig proteína/mL, com tempo de cultivo de 72 horas e com 24 horas de incubação com IgG. A massa molecular da fração ativa de protease foi compreendida entre 43 a 150 kDa.

  4. Actinobacillus Actinomycetemcomitans y Porphyromonas Gingivales como principales patógenos periodontales

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    A Bascones

    2000-09-01

    Full Text Available Entre las bacterias relacionadas con la enfermedad periodontal, existen dos especies más claramente asociadas a esta enfermedad: Actinobacillus actinomycetemcomitans y Porphyromonas gingivalis. Este trabajo es una revisión bibliográfica sobre estos dos patógenos periodontales, mostrando su origen, prevalencia, distribución, transmisión y respuesta al tratamiento periodontal.Among the bacteria related to periodontal disease, there are two species clearly associated to this disease: Actinobacillus actinomycetemcomitans and Porphyromonas gingiva lis. This paper presents a review of the literature regarding this two periodontal pathogens, and showing their origin, prevalence, distribution, transmission and response to periodontal treatment.

  5. Improved, low-cost selective culture medium for Actinobacillus actinomycetemcomitans.

    Science.gov (United States)

    Alsina, M; Olle, E; Frias, J

    2001-02-01

    Actinobacillus actinomycetemcomitans is considered to be one of the major oral putative pathogens, especially in cases of juvenile periodontitis. This microorganism requires nutritionally complex media for growth, and therefore the media for its primary isolation usually include blood agar or serum in their base. In this study we present a new medium, Dentaid-1, which improves the detection of A. actinomycetemcomitans in periodontal samples. In its composition, blood and serum have been omitted, hence reducing its cost and making it a more restrictive medium against the growth of other microorganisms with high nutritional requirements. The growth yields of pure cultures of the bacteria on Dentaid-1 were comparable to those on nonselective blood agar. Moreover, clinical efficacy was evaluated in subgingival samples from 77 subjects with adult periodontitis. Dentaid-1 detected A. actinomycetemcomitans in 24 subjects, while a previously described tryptic soy-serum-bacitracin-vancomycin agar detected the microorganism in only 19 subjects (79.1%). Dentaid-1 is a low-cost, noninhibitory formula for the improved diagnosis and monitoring of patients subgingivally infected by this important oral putative pathogen.

  6. Oxidative and nonoxidative killing of Actinobacillus actinomycetemcomitans by human neutrophils.

    Science.gov (United States)

    Miyasaki, K T; Wilson, M E; Brunetti, A J; Genco, R J

    1986-07-01

    Actinobacillus actinomycetemcomitans is a facultative gram-negative microorganism which has been implicated as an etiologic agent in localized juvenile periodontitis and in subacute bacterial endocarditis and abscesses. Although resistant to serum bactericidal action and to oxidant injury mediated by superoxide anion (O2-) and hydrogen peroxide (H2O2), this organism is sensitive to killing by the myeloperoxidase-hydrogen peroxide-chloride system (K.T. Miyasaki, M.E. Wilson, and R.J. Genco, Infect. Immun. 53:161-165, 1986). In this study, we examined the sensitivity of A. actinomycetemcomitans to killing by intact neutrophils under aerobic conditions, under anaerobic conditions, and under aerobic conditions in the presence of the heme-protein inhibitor sodium cyanide. Intact neutrophils killed opsonized A. actinomycetemcomitans under aerobic and anaerobic conditions, and the kinetics of these reactions indicated that both oxidative and nonoxidative mechanisms were operative. Oxidative mechanisms contributed significantly, and most of the killing attributable to oxidative mechanisms was inhibited by sodium cyanide, which suggested that the myeloperoxidase-hydrogen peroxide-chloride system participated in the oxidative process. We conclude that human neutrophils are capable of killing A. actinomycetemcomitans by both oxygen-dependent and oxygen-independent pathways, and that most oxygen-dependent killing requires myeloperoxidase activity.

  7. Catecholamines promote Actinobacillus pleuropneumoniae growth by regulating iron metabolism.

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    Lu Li

    Full Text Available Catecholamines are host stress hormones that can induce the growth of many bacteria by facilitating iron utilization and/or regulate the expression of virulence genes through specific hormone receptors. Whether these two responsive pathways are interconnected is unknown. In our previous study, it was found that catecholamines can regulate the expression of a great number of genes of Actinobacillus pleuropneumoniae, an important swine respiratory pathogen. However, bacterial growth was not affected by catecholamines in rich medium. In this study, it was discovered that catecholamines affected A. pleuropneumoniae growth in chemically defined medium (CDM. We found that serum inhibited A. pleuropneumoniae growth in CDM, while epinephrine, norepinephrine and dopamine promoted A. pleuropneumoniae growth in the CDM containing serum. The known bacterial hormone receptor QseC didn't play roles in this process. Ion-supplementation and transcriptome analysis indicated that serum addition resulted in iron-restricted conditions which were alleviated by the addition of catecholamines. Transferrin, one of the components in serum, inhibited the growth of A. pleuropneumoniae in CDM, an effect reversed by addition of catecholamines in a TonB2-dependent manner. Our data demonstrate that catecholamines promote A. pleuropneumoniae growth by regulating iron-acquisition and metabolism, which is independent of the adrenergic receptor QseC.

  8. Catecholamines promote Actinobacillus pleuropneumoniae growth by regulating iron metabolism.

    Science.gov (United States)

    Li, Lu; Chen, Zhaohui; Bei, Weicheng; Su, Zhipeng; Huang, Qi; Zhang, Liang; Chen, Huanchun; Zhou, Rui

    2015-01-01

    Catecholamines are host stress hormones that can induce the growth of many bacteria by facilitating iron utilization and/or regulate the expression of virulence genes through specific hormone receptors. Whether these two responsive pathways are interconnected is unknown. In our previous study, it was found that catecholamines can regulate the expression of a great number of genes of Actinobacillus pleuropneumoniae, an important swine respiratory pathogen. However, bacterial growth was not affected by catecholamines in rich medium. In this study, it was discovered that catecholamines affected A. pleuropneumoniae growth in chemically defined medium (CDM). We found that serum inhibited A. pleuropneumoniae growth in CDM, while epinephrine, norepinephrine and dopamine promoted A. pleuropneumoniae growth in the CDM containing serum. The known bacterial hormone receptor QseC didn't play roles in this process. Ion-supplementation and transcriptome analysis indicated that serum addition resulted in iron-restricted conditions which were alleviated by the addition of catecholamines. Transferrin, one of the components in serum, inhibited the growth of A. pleuropneumoniae in CDM, an effect reversed by addition of catecholamines in a TonB2-dependent manner. Our data demonstrate that catecholamines promote A. pleuropneumoniae growth by regulating iron-acquisition and metabolism, which is independent of the adrenergic receptor QseC.

  9. Salivary lactoferrin and low-M-r mucin MG2 in Actinobacillus actinomycetemcomitans-associated periodontitis

    NARCIS (Netherlands)

    Groenink, J; Walgreen-Weterings, E; Nazmi, K; Bolscher, JGM; Veerman, ECI; van Winkelhoff, AJ; Amerongen, AVH

    1999-01-01

    Concentrations and output of lactoferrin and of low-M-r mucin MG2 were determined in saliva of subjects suffering from Actinobacillus actinomycetem-comitans-associated periodontal disease and healthy subjects. Periodontal patients were clinically examined and a microbiological sample was taken from

  10. Transmission of Actinobacillus pleuropneumoniae among weaned piglets on endemically infected farms

    NARCIS (Netherlands)

    Tobias, T.J.; Bouma, A.; Broek, van den J.; Nes, van A.; Daemen, A.J.J.M.; Wagenaar, J.A.; Stegeman, J.A.; Klinkenberg, D.

    2014-01-01

    Clinical outbreaks due to Actinobacillus pleuropneumoniae occur recurrently, despite the wide-scale use of antimicrobials or vaccination. Therefore, new approaches for the prevention and control of these outbreaks are necessary. For the development of alternative measures, more insight into the tran

  11. Early-onset periodontitis in Morocco is associated with the highly leukotoxic clone of Actinobacillus actinomycetemcomitans

    DEFF Research Database (Denmark)

    Haubek, Dorte; Ennibi, O.-K.; Poulsen, Knud

    2001-01-01

    A particular clone (JP2) of Actinobacillus actinomycetemcomitans with increased leukotoxin production has been isolated from individuals with early-onset periodontitis (EOP). The aim of this study was to determine the frequency of carriers of this clone and its association with EOP in Moroccan...

  12. Actinobacillus pleuropneumoniae osteomyelitis in pigs demonstrated by fluorescent in situ hybridization

    DEFF Research Database (Denmark)

    Jensen, Tim Kåre; Boye, Mette; Hagedorn-Olsen, T.;

    1999-01-01

    Necrotizing osteomyelitis and fibrinopurulent arthritis with isolation of Actinobacillus pleuropneumoniae serotype 2 is reported in two pigs from a herd with lameness and mild coughing problems among 8 to 12-week-old pigs. Application of fluorescent in situ hybridization targeting 16S ribosomal RNA...

  13. The interaction between saliva and Actinobacillus actinomycetemcomitans influenced by the Zeta potential

    NARCIS (Netherlands)

    Groenink, J; Veerman, ECI; Zandvoort, MS; Van der Mei, HC; Busscher, HJ; Amerongen, AVN

    1998-01-01

    The adhesion of Actinobacillus actinomycetemcomitans is a virulence factor in the aetiology of periodontitis and is determined by physico-chemical properties, e.g. surface charge and hydrophobicity, of the bacterial cell surface. Although oral surfaces are constantly coated with saliva, few studies

  14. Transcriptional profiling of Actinobacillus pleuropneumoniae under iron-restricted conditions

    Directory of Open Access Journals (Sweden)

    Harel Josée

    2007-03-01

    Full Text Available Abstract Background To better understand effects of iron restriction on Actinobacillus pleuropneumoniae and to identify new potential vaccine targets, we conducted transcript profiling studies using a DNA microarray containing all 2025 ORFs of the genome of A. pleuropneumoniae serotype 5b strain L20. This is the first study involving the use of microarray technology to monitor the transcriptome of A. pleuropneumoniae grown under iron restriction. Results Upon comparing growth of this pathogen in iron-sufficient versus iron-depleted medium, 210 genes were identified as being differentially expressed. Some genes (92 were identified as being up-regulated; many have confirmed or putative roles in iron acquisition, such as the genes coding for two TonB energy-transducing proteins and the hemoglobin receptor HgbA. Transcript profiling also led to identification of some new iron acquisition systems of A. pleuropneumoniae. Genes coding for a possible Yfe system (yfeABCD, implicated in the acquisition of chelated iron, were detected, as well as genes coding for a putative enterobactin-type siderophore receptor system. ORFs for homologs of the HmbR system of Neisseria meningitidis involved in iron acquisition from hemoglobin were significantly up-regulated. Down-regulated genes included many that encode proteins containing Fe-S clusters or that use heme as a cofactor. Supplementation of the culture medium with exogenous iron re-established the expression level of these genes. Conclusion We have used transcriptional profiling to generate a list of genes showing differential expression during iron restriction. This strategy enabled us to gain a better understanding of the metabolic changes occurring in response to this stress. Many new potential iron acquisition systems were identified, and further studies will have to be conducted to establish their role during iron restriction.

  15. Identification of Actinobacillus pleuropneumoniae serotypes 1, 7, and 12 by multiplex PCR based on genes involved in encapsulation

    DEFF Research Database (Denmark)

    Angen, Øystein; Jessing, Stine Graakjær; Ahrens, Peter;

    2005-01-01

    Based on differences in the capsular polysaccharides, 15 serotypes have until now been described for Actinobacillus pleuropneumoniae, the etiological agent of swine pleuropneumonia. Identification of the causative serotype is important both as a virulence marker and for epidemiological purposes. ...

  16. Molecular characterisation of the early response in pigs to experimental infection with Actinobacillus pleuropneumoniae using cDNA microarrays

    DEFF Research Database (Denmark)

    Hedegaard, Jakob; Skovgaard, Kerstin; Mortensen, Shila;

    2007-01-01

    Background: The bacterium Actinobacillus pleuropneumoniae is responsible for porcine pleuropneumonia, a widespread, highly contagious and often fatal respiratory disease of pigs. The general porcine innate immune response after A. pleuropneumoniae infection is still not clarified. The objective o...

  17. Multiplex PCR that can distinguish between immunologically cross-reactive serovar 3, 6, and 8 Actinobacillus pleuropneumoniae strains

    DEFF Research Database (Denmark)

    Zhou, L.; Jones, S.C.P.; Angen, Øystein;

    2008-01-01

    We describe a highly sensitive and specific multiplex PCR, based on capsular loci and the species specific apxIV gene, that unequivocally differentiates serovar 3, 6, and 8 Actinobacillus pleuropneumoniae strains that are cross-reactive in conventional immunological tests.......We describe a highly sensitive and specific multiplex PCR, based on capsular loci and the species specific apxIV gene, that unequivocally differentiates serovar 3, 6, and 8 Actinobacillus pleuropneumoniae strains that are cross-reactive in conventional immunological tests....

  18. Experimental model of swine pneumonic pasteurellosis using crude Actinobacillus pleuropneumoniae cytotoxin and Pasteurella multocida given endobronchially.

    OpenAIRE

    Chung, W. B.; Bäckström, L R; Collins, M T

    1994-01-01

    This study was designed to develop and characterize a swine pneumonic pasteurellosis model by concurrent introduction of Pasteurella multocida type A and Actinobacillus pleuropneumoniae crude cytotoxin. After a series of preliminary experiments, a combination of 4 x 10(9) P. multocida and 4,000 toxic units of A. pleuropneumoniae crude cytotoxin was determined to produce optimal results. A total of 48 pigs were divided into four groups of 12 pigs each. The control group received buffered salin...

  19. Actinobacillus pleuropneumoniae culture supernatants interfere with killing of Pasteurella multocida by swine pulmonary alveolar macrophages.

    OpenAIRE

    Chung, W. B.; Bäckström, L; McDonald, J.; Collins, M T

    1993-01-01

    The effect of Actinobacillus pleuropneumoniae culture supernatant on swine pulmonary alveolar macrophage (PAM) functions was studied. The A. pleuropneumoniae culture supernatant was toxic to PAMs when tested by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and lactate dehydrogenase (LDH) release assays. Biological activity of the supernatant was ascribed to cytotoxins. Both the LDH and MTT assays were used for measurement of crude A. pleuropneumoniae cytotoxin concentrati...

  20. Growth of Actinobacillus pleuropneumoniae is promoted by exogenous hydroxamate and catechol siderophores.

    OpenAIRE

    Diarra, M. S.; Dolence, J A; Dolence, E K; Darwish, I; Miller, M.J.; Malouin, F; Jacques, M.

    1996-01-01

    Siderophores bind ferric ions and are involved in receptor-specific iron transport into bacteria. Six types of siderophores were tested against strains representing the 12 different serotypes of Actinobacillus pleuropneumoniae. Ferrichrome and bis-catechol-based siderophores showed strong growth-promoting activities for A. pleuropneumoniae in a disk diffusion assay. Most strains of A. pleuropneumoniae tested were able to use ferrichrome (21 of 22 or 95%), ferrichrome A (20 of 22 or 90%), and ...

  1. ohr, Encoding an Organic Hydroperoxide Reductase, Is an In Vivo-Induced Gene in Actinobacillus pleuropneumoniae

    OpenAIRE

    Shea, Robin J.; Mulks, Martha H.

    2002-01-01

    Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia, a disease characterized by pulmonary necrosis and hemorrhage caused in part by neutrophil degranulation. In an effort to understand the pathogenesis of this disease, we have developed an in vivo expression technology (IVET) system to identify genes that are specifically up-regulated during infection. One of the genes that we have identified as being induced in vivo is ohr, encoding organic hydroperoxide reducta...

  2. Cellular and molecular responses of periodontal connective tissue cells to Actinobacillus actinomycetemcomitans cytolethal distending toxin

    OpenAIRE

    2004-01-01

    Actinobacillus actinomycetemcomitans is present in elevated proportions and numbers in dental bacterial biofilms of patients with localized aggressive periodontitis. This variant of periodontal disease, occurring in adolescents and young adults, is characterized by rapid and severe destruction of the connective tissues and bone supporting the teeth, eventually culminating in tooth loss. The cytolethal distending toxin (Cdt) is a newly discovered bacterial protein toxin, uniquely present in A....

  3. Actinobacillus suis and Actinobacillus equuli, emergent pathogens of septic embolic nephritis, a new challenge for the swine industry Actinobacillus suis y Actinobacillus equuli, patógenos emergentes de nefritis embólica séptica, un nuevo desafío para la industria porcina

    Directory of Open Access Journals (Sweden)

    CE Benavente

    2012-01-01

    Full Text Available Kidney lesions are an important cause of tissue condemnation in slaughterhouses. In addition to the potential public health implications, organ condemnations have a significant economic impact on the food animal industry. The condition classified broadly as "nephritis" is one of the main causes of tissue condemnation. Embolic nephritis resembling Actinobacillus equuli infection in foals has been recently detected in sows and market hogs. Actinobacillus suis is phenotypically and phylogenetically closely related to A. equuli. Both are Gram-negative bacteria, not easy to detect in routine exams. A. suis is an opportunistic pathogen that can produce fatal septicaemia in pigs, pneumonia, polyarthritis, septic embolic nephritis, abortion and mummified foetuses. Outbreaks of clinical disease appear to occur more frequently in high-health-status herds. In adult pigs the skin lesions may be confused with porcine erysipelas. A. suis and A. equuli are emerging opportunistic pathogens in the porcine industry and both have potential public health consequences to people that handles meat products. The objective of this paper is to present a literature review regarding the role of A. suis and A. equuli in the pathogenesis of nephritis in swine.Las lesiones renales son una causa importante de decomiso en los mataderos. Además de las posibles consecuencias en salud pública, el decomiso de órganos tiene un gran impacto económico en la industria de alimento animal. Recientemente, nefritis embólica séptica con lesiones semejantes a infecciones con Actinobacillus equuli en potrillos ha sido detectada en reproductoras y cerdos con peso de mercado. Actinobacillus equuli es fenotípica y genéticamente similar a Actinobacillus suis. Ambas son bacterias Gram-negativas difíciles de diagnosticar en exámenes de rutina. A. suis es un patógeno oportunista capaz de producir septicemia en cerdos, neumonía, poliartritis, nefritis embólica séptica, aborto y fetos

  4. Haemolytic activity of Actinobacillus actinomycetemcomitans strains on different blood types Atividade hemolítica de cepas de Actinobacillus actinomycetemcomitans em diferentes tipos sanguíneos

    Directory of Open Access Journals (Sweden)

    Mario Julio Avila-Campos

    1995-06-01

    Full Text Available Haemolytic activity of sixty nine Actinobacillus actinomycetemcomitans strains on different animal and human blood types was examined by using a trypticase soy agar supplemented with yeast extract (0.5%. Blood types used were: rabbit, sheep and human (A, Rh+; A, Rh-; B, Rh+; B, Rh-; O, Rh+; O, Rh-; AB, Rh+; AB, Rh- groups. Plates were inoculated and, incubated in microaerophilic conditions, at 37ºC, for 48 h. The haemolytic activity of the tested strains was characterized as alpha-haemolysis. Only two isolates were not haemolytic on all blood types (2.9%, two strains were haemolytic only on human blood (one strain on AB, Rh+ group and another one on A, Rh+ and AB, Rh+ groups. No specificity between haemolysin produced by the tested strains and blood type was observed.A atividade hemolítica de 69 cepas de Actinobacillus actinomycetemcomitans foi determinada em diferentes tipos de sangue animal e humano, usando como meio base ágar de soja tripticaseina, suplementado com extrato de levedura (0,5%. Foram utilizados sangue de coelho, carneiro e humano (grupos A, Rh+; A, Rh-; B, Rh+; B, Rh-; O, Rh+; O, Rh-; AB, Rh+ e AB, Rh-. As placas foram inoculadas e, incubadas em condições de microaerofilia, a 37ºC, por 48 h. A atividade hemolítica das cepas testadas foi caracterizada como alfa-hemólise. Somente dois (2,9% isolados não hemolisaram todos os tipos sanguíneos, duas cepas hemolisaram somente sangue humano (uma o grupo AB, Rh+ e outra os grupos A, Rh+ e AB, Rh+. Não foi observada alguma especificidade entre as hemolisinas produzidas e os tipos de sangue utilizados.

  5. Detection of Actinobacillus pleuropneumoniae in pigs by real-time quantitative PCR for the apxIVA gene

    NARCIS (Netherlands)

    Tobias, T.J.; Bouma, A.; Klinkenberg, D.; Daemen, A.J.J.M.; Stegeman, J.A.; Wagenaar, J.A.; Duim, B.

    2012-01-01

    A real-time quantitative PCR (qPCR) for detection of the apxIVA gene of Actinobacillus pleuropneumoniae was validated using pure cultures of A. pleuropneumoniae and tonsillar and nasal swabs from experimentally inoculated Caesarean-derived/colostrum-deprived piglets and naturally infected convention

  6. Blocking enzyme-linked immunosorbent assay for detection of antibodies against Actinobacillus pleuropneumoniae serotype 6 in pig serum

    DEFF Research Database (Denmark)

    Klausen, Joan; Andresen, Lars Ole; Barfod, Kristen

    2001-01-01

    A blocking enzyme-linked immunosorbent assay (ELISA) detecting antibodies against Actinobacillus pleuropneumoniae (Ap) serotype 6 was developed. The blocking ELISA was based on the inhibition of a polyclonal antibody raised against Ap serotype 6. Purified lipopolysaccharide from Ap serotype 6...

  7. Changes in antimicrobial susceptibility of Actinobacillus pleuropneumoniae isolated from pigs in Spain during the last decade.

    Science.gov (United States)

    Gutiérrez-Martín, César B; del Blanco, Noemí García; Blanco, Mónica; Navas, Jesús; Rodríguez-Ferri, Elías F

    2006-06-15

    A total of 229 Spanish Actinobacillus pleuropneumoniae isolates recovered from diseased pigs with pleuropneumonia from 1997 to 2004 was tested for their susceptibility to 11 antimicrobials in a broth microdilution method. All the isolates were susceptible to florfenicol and most of them to cephalothin; however, a high rate of resistance was observed to tetracycline. A bimodal or multimodal distribution of isolates over the MIC range were observed for penicillins, tetracycline, trimethoprim, sulfisoxazole and nalidixic acid, suggesting the development of acquired resistance. Eight resistance patterns were established, and 21.1% of the isolates were resistant to at least two antimicrobials. In addition, a considerable increase in the resistance to tetracyclines was observed during the last decade in Spain, when compared with other A. pleuropneumoniae strains isolated during 1987-1988 (Gutiérrez, C.B., Píriz, S., Vadillo, S., Rodríguez Ferri, E.F., 1993. In vitro susceptibility of Actinobacillus pleuropneumoniae strains to 42 antimicrobial agents. Am. J. Vet. Res. 54, 546-550); this finding was also observed for gentamicin in minor percentage.

  8. Actinobacillus pleuropneumoniae serotype 10 derived ApxI induces apoptosis in porcine alveolar macrophages.

    Science.gov (United States)

    Chien, Maw-Sheng; Chan, You-Yu; Chen, Zeng-Weng; Wu, Chi-Ming; Liao, Jiunn-Wang; Chen, Ter-Hsin; Lee, Wei-Cheng; Yeh, Kuang-Sheng; Hsuan, Shih-Ling

    2009-03-30

    Actinobacillus pleuropneumoniae (AP) is the causative agent of swine pleuropneumonia, a fibrinous, exudative, hemorrhagic, necrotizing pleuropneumonia affecting all ages of pigs. Actinobacillus pleuropneumoniae exotoxins (Apx) are one of the major virulence factors of AP. Due to the complex nature of Apx toxins produced by AP, little is known regarding the interactions of individual species of Apx toxin with target cells. The objective of this study was to examine whether AP serotype 10-derived exotoxin, ApxI, caused apoptosis in porcine alveolar macrophages (PAMs) and to delineate the underlying signaling pathways. Isolated PAMs were stimulated with different concentrations of native ApxI and monitored for apoptosis using Hoechst staining, TUNEL, and DNA laddering assays. The ApxI-stimulated PAMs exhibited typical morphological features of apoptosis, including condensation of chromatin, formation of apoptotic bodies and DNA laddering. ApxI-induced apoptosis in a concentration- and time-dependent manner. Furthermore, to delineate the signaling events involved in ApxI-induced apoptosis, it was observed that caspase 3 was activated in ApxI-stimulated PAMs. Ablation of caspase 3 activity via specific inhibitors protected PAMs from apoptosis by ApxI. This study is the first to demonstrate that native ApxI causes apoptosis in PAMs at low concentrations and that these apoptotic events are mediated via a caspase 3-dependent pathway. These findings suggest a role of ApxI in AP infection as it might impair the host defense system through the induction of apoptosis in PAMs.

  9. Production of succinic acid from oil palm empty fruit bunch cellulose using Actinobacillus succinogenes

    Science.gov (United States)

    Pasma, Satriani Aga; Daik, Rusli; Maskat, Mohamad Yusof

    2013-11-01

    Succinic acid is a common metabolite in plants, animals and microorganisms. It has been used widely in agricultural, food and pharmaceutical industries. Enzymatic hydrolysate glucose from oil palm empty fruit bunch (OPEFB) cellulose was used as a substrate for succinic acid production using Actinobacillus succinogenes. Using cellulose extraction from OPEFB can enhance the production of glucose as a main substrate for succinic acid production. The highest concentration of glucose produced from enzymatic hydrolysis is 167 mg/mL and the sugar recovery is 0.73 g/g of OPEFB. By optimizing the culture medium for succinic acid fermentation with enzymatic hydrolysate of OPEFB cellulose, the nitrogen sources could be reduced to just only 2.5 g yeast extract and 2.5 g corn step liquor. Batch fermentation was carried out using enzymatic hydrolysate of OPEFB cellulose with yeast extract, corn steep liquor and the salts mixture, 23.5 g/L succinic acid was obtained with consumption of 72 g/L glucose in enzymatic hydrolysate of OPEFB cellulose at 38 hours and 37°C. This study suggests that enzymatic hydrolysate of OPEFB cellulose maybe an alternative substrate for the efficient production of succinic acid by Actinobacillus succinogenes.

  10. Virulence factors of Actinobacillus actinomycetemcomitans: other putative factors Fatores de virulência do Actinobacillus actinomycetemcomitans: outros possíveis fatores

    Directory of Open Access Journals (Sweden)

    Mario Julio AVILA-CAMPOS

    2000-03-01

    Full Text Available Actinobacillus actinomycetemcomitans is implicated as the causative agent of localized juvenile periodontitis. This organism possesses a large number of virulence factors with a wide range of activities and also interfere with tissue repair. Fifty isolates of A. actinomycetemcomitans from 20 periodontal patients were examined to evaluate other putative virulence factors. In this study, the capsule, DNase, coagulase, fibrinolysin, proteolytic, haemolysin and bacteriocin production, haemagglutination, serum sensitivity, epithelial cells attachment, hydrophobicity and virulence of the A. actinomycetemcomitans isolates were evaluated. All the isolates were resistant to the different tested sera. 70% to 94% were alpha-haemolytics and agglutinated all blood types. Most of isolates produced antagonistic substances and they had a low hydrophobicity. None of the isolates was pathogenic for mice. Little is known as to wether these factors may act in the development of periodontal disease, and further studies are required for an application in pathogenic and systematic terms.Actinobacillus actinomycetemcomitans está implicado como o agente etiológico da periodontite juvenil localizada. Este organismo possui inúmeros fatores de virulência que podem interferir no reparo tissular. 50 isolados de A. actinomycetemcomitans de pacientes com periodontite foram examinados para avaliar outros possíveis fatores de virulência. Neste estudo, foi avaliada a produção de cápsula, DNase, coagulase, fibrinolisina, atividade proteolítica, hemolisina e bacteriocina, assim como hemaglutinação, sensibilidade ao soro, aderência às células epiteliais, hidrofobicidade e virulência de A. actinomycetemcomitans. Todos os isolados foram resistentes para todos os tipos de soro utilizados. 70% a 94% dos isolados foram alfa-hemolíticos e aglutinaram todos os tipos sanguíneos. A maioria dos isolados produziu substâncias antagonistas e apresentaram baixa hidrofobicidade

  11. Alterações patológicas em potros infectados por Actinobacillus equuli subsp. haemolyticus Pathological changes in foals infected with Actinobacillus equuli subsp. haemolyticus

    Directory of Open Access Journals (Sweden)

    Danilo Carloto Gomes

    2010-06-01

    Full Text Available Neste trabalho, são descritos dois casos fatais de septicemia com lesões embólicas causadas por Actinobacillus equuli subsp. haemolyticus em potros recém-nascidos. Em um dos animais, foram observados, na necropsia, pequenos nódulos esbranquiçados de aproximadamente 0,2cm de diâmetro na cortical dos rins e no outro havia uma área de coloração acinzentada no lobo diafragmático esquerdo do pulmão. As principais alterações microscópicas observadas no primeiro animal foram rins com infiltrado inflamatório multifocal a coalescente acentuado, com predomínio de neutrófilos, associado com áreas basofílicas levemente granulares compostas por grumos bacterianos. No segundo animal, o pulmão apresentava infiltrado inflamatório neutrofílico, edema, congestão e colônias bacterianas intravasculares. Em ambos os casos, colônias bacterianas foram encontradas disseminadas por vários órgãos incluindo capilares cerebrais. Nos dois casos foi isolado e identificado A. equuli subsp. haemolyticus.This paper describes two fatal cases of embolic and septicaemic lesions caused by Actinobacillus equuli subsp. haemolyticus in two newborn foals. In one foal was observed at necropsy small whitish nodules of approximately 0,2cm in diameter on the renal cortex and the other foal had an area of gray color in the left diaphragmatic lobe of the lung. The main histologic changes were observed in the first foal kidneys with multifocal to coalescing inflammatory suppurative infiltrates associated with slightly granular basophilic bacterial colonies. In the second animal the lung showed neutrophilic inflammatory infiltrate, edema, congestion and presence of intravascular bacterial colonies. In both cases, the bacteria were disseminated by several organs including cerebral capillary cerebral. In both cases A. equuli subsp. haemolyticus was isolated and identified.

  12. Serotyping of Actinobacillus pleuropneumoniae serotype 5 strains using a monoclonal-based polystyrene agglutination test

    DEFF Research Database (Denmark)

    Dubreuil, J.D.; Letellier, A.; Stenbæk, Eva;

    1996-01-01

    A polystyrene agglutination test has been developed for serotyping Actinobacillus pleuropneumoniae serotype 5a and 5b strains. Protein A-coated polystyrene microparticles were sensitized with a murine monoclonal antibody recognizing an epitope on serotype 5 LPS-O chain as shown by SDS......-PAGE and Western blotting, A total of 205 A. pleuropneumoniae, strains including all 12 serotype reference strains and 13 strains representing 8 common bacterial species associated with swine or related to A, pleuropneumoniae, were tested by mixing 25 mu L of polystyrene reagent with the same volume of a dense...... suspension of bacterial cells grown for 18 h. All A, pleuropneumoniae strains had been previously serotyped using standard procedures, The polystyrene agglutination test was rapid (less than 3 min) and easy to perform. Overall a very good correlation (97.3%) with the standard techniques was found...

  13. Cloning, Expression of apxI Gene of Actinobacillus pleuropneumoniae and Development of ELISA

    Institute of Scientific and Technical Information of China (English)

    LIU Jian-jie; HE Qi-gai; CHEN Huan-chun; WU Bin; XU Xiao-juan; LIU Jun-fa; TANG Xian-chun; BEI Wei-cheng

    2003-01-01

    Based on the published nucleotide sequence of the apxICA of Actinobacillus pleuropneumoniaein Genbank(S4074), a pair of primers were designed. A 3 640 bp(4 687 -8 326 bp)gene fragment was ampli-fied by PCR from the isolated strain of A. pleuropneumoniae serovar 1. Then, it was cloned into pMD18-T,identified by both restriction endonuclease and sequence analysis, and inserted into pET-28a expression vectorto yield the expression plasmid. SDS-PAGE result indicated expression of apxICA in BL21 (DE3), Westernblot analysis showed the protein's immunogenicity. Using the expressed protein, ELISA was established to de-tect serum antibody against ApxI. The feature of ELISA to detect highly virulent A. pleuropneumoniae strainsinfection was proved by primary clinical application.

  14. Isolation of Actinobacillus seminis from a goat with clinical epididymo-orchitis in Brazil.

    Science.gov (United States)

    dos Santos, Fabrine Alexandre; de Azevedo, Edísio Oliveira; de Azevedo, Sérgio Santos; Garino Júnior, Felício; Mota, Rinaldo Aparecido; de Cássia Peixoto Kim, Pomy; Gomes, Ana Lisa Vale; Alves, Clebert José

    2014-01-01

    The present study reports the first isolation of Actinobacillus seminis from a goat in Brazil. A four-year-old Moxotó breeding goat in a flock of 70 goats and 65 sheep reared together in the county of Patos, semiarid region of Northeastern Brazil, showed clinical signs of unilateral orchitis and epididymitis. Diagnosis of A. seminis infection was confirmed by association of clinical findings, bacterial isolation and 16S rRNA gene sequencing. This result suggests that A. seminis may be an additional cause of infertility in goats, and that sheep may be the source of infection because the mixed farming system allows the contact between sheep and goats in the semiarid region of Northeastern Brazil.

  15. Isolation of Actinobacillus seminis from a goat with clinical epididymo-orchitis in Brazil

    Directory of Open Access Journals (Sweden)

    Fabrine Alexandre dos Santos

    2014-01-01

    Full Text Available The present study reports the first isolation of Actinobacillus seminis from a goat in Brazil. A four-year-old Moxotó breeding goat in a flock of 70 goats and 65 sheep reared together in the county of Patos, semiarid region of Northeastern Brazil, showed clinical signs of unilateral orchitis and epididymitis. Diagnosis of A. seminis infection was confirmed by association of clinical findings, bacterial isolation and 16S rRNA gene sequencing. This result suggests that A. seminis may be an additional cause of infertility in goats, and that sheep may be the source of infection because the mixed farming system allows the contact between sheep and goats in the semiarid region of Northeastern Brazil.

  16. Succinic acid production from corn stover by simultaneous saccharification and fermentation using Actinobacillus succinogenes.

    Science.gov (United States)

    Zheng, Pu; Fang, Lin; Xu, Yan; Dong, Jin-Jun; Ni, Ye; Sun, Zhi-Hao

    2010-10-01

    Simultaneous saccharification and fermentation (SSF) technique was applied for succinic acid production by Actinobacillus succinogenes in a 5-l stirred bioreactor with corn stover as the raw material. The process parameters of SSF, including corn stover pretreatment condition, substrate concentration, enzyme loading and fermentation temperature were investigated. Results indicated that pretreating corn stover with diluted alkaline was beneficial for the succinic acid production, and succinic acid yield could be significantly increased when adding the cellulase supplemented with cellobiase. The maximal succinic acid concentration and yield could reach 47.4 g/l and 0.72 g/g-substrate, respectively. The corresponding operation conditions were summarized as follows: SSF operation at 38 °C for 48 h, diluted alkaline pretreated corn stover as substrate with concentration of 70 g/l, enzyme loading of 20FPU cellulase and 10 U cellobiase per gram substrate. This result suggested an industrial potential of succinic acid production by using SSF and corn stover.

  17. Prevalence of leukotoxic genotypes of Actinobacillus actinomycetemcomitans in Brazilians with chronic periodontitis Prevalência do genotipo leucotóxico de Actinobacillus actinomycetemcomitans em indivíduos brasileiros com periodontite crônica

    OpenAIRE

    Wilson Rosalem Junior; Arnaldo Feitosa Braga de Andrade; Ana Paula Vieira Colombo

    2006-01-01

    Actinobacillus actinomycetemcomitans is considered a major etiologic agent of aggressive periodontitis but this species has also been associated with other forms of periodontal disease. Further, highly leukotoxic strains are related to severity of disease. This investigation determined the prevalence of A. actinomycetemcomitans and the occurrence of the leukotoxin gene 530-bp deletion in Brazilian subjects with chronic periodontitis. Twenty periodontally healthy and 20 chronic periodontitis s...

  18. Comparison of conventional and long-acting oxytetracyclines in prevention of induced Actinobacillus (Haemophilus) pleuropneumoniae infection of growing swine.

    OpenAIRE

    Kiorpes, A L; Bäckström, L R; Collins, M T; Kruse, G O

    1989-01-01

    These experiments tested the hypothesis that long-acting oxytetracycline (oxytetracycline-LA) was more effective than regular oxytetracycline in preventing porcine pleuropneumonia when administered either 24 or 48 h prior to experimental challenge with virulent strains of Actinobacillus pleuropneumoniae. Two experiments (1 and 2) were conducted using growing pigs (average weight 12-15 kg). Antibiotic treatments were administered once intramuscularly at 20 mg/kg body weight; controls received ...

  19. Differential effect of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans on co-cultures of human oral cells

    OpenAIRE

    Kang, Philip; Korostoff, Jonathan; Volgina, Alla; Grzesik, Wojciech; DiRienzo, Joseph M.

    2005-01-01

    The periodontal pathogen Actinobacillus actinomycetemcomitans expresses a cytolethal distending toxin (CDT) that typically arrests the growth of eukaryotic cells at either the G0/G1 or G2/M phase of the cell cycle. It was previously found that CDT failed to arrest the growth of human periodontal ligament fibroblasts (HPLFs) when grown in pure culture. In contrast, proliferation of an oral epithelial cell line was rapidly inhibited by the toxin. In this study, the feasibility of using mixed-ce...

  20. Branched-Chain Amino Acids Are Required for the Survival and Virulence of Actinobacillus pleuropneumoniae in Swine▿

    OpenAIRE

    Subashchandrabose, Sargurunathan; LeVeque, Rhiannon M.; Wagner, Trevor K.; Kirkwood, Roy N; Kiupel, Matti; Mulks, Martha H.

    2009-01-01

    In Actinobacillus pleuropneumoniae, which causes porcine pleuropneumonia, ilvI was identified as an in vivo-induced (ivi) gene and encodes the enzyme acetohydroxyacid synthase (AHAS) required for branched-chain amino acid (BCAA) biosynthesis. ilvI and 7 of 32 additional ivi promoters were upregulated in vitro when grown in chemically defined medium (CDM) lacking BCAA. Based on these observations, we hypothesized that BCAA would be found at limiting concentrations in pulmonary secretions and t...

  1. Transcriptional Profiling of Hilar Nodes from Pigs after Experimental Infection with Actinobacillus Pleuropneumoniae

    Directory of Open Access Journals (Sweden)

    Shumin Yu

    2013-11-01

    Full Text Available The gram-negative bacterium Actinobacillus pleuropneumoniae (APP is an inhabitant of the porcine upper respiratory tract and the causative agent of porcine pleuropneumonia (PP. In recent years, knowledge about the proinflammatory cytokine and chemokine gene expression that occurs in lung and lymph node of the APP-infected swine has been advanced. However, systematic gene expression profiles on hilar nodes from pigs after infection with Actinobacillus pleuropneumoniae have not yet been reported. The transcriptional responses were studied in hilar nodes (HN from swine experimentally infected with APP and the control groupusing Agilent Porcine Genechip, including 43,603 probe sets. 9,517 transcripts were identified as differentially expressed (DE at the p ≤ 0.01 level by comparing the log2 (normalized signal of the two groups named treatment group (TG and controls (CG. Eight hundred and fifteen of these DE transcripts were annotated as pig genes in the GenBank database (DB. Two hundred and seventy-two biological process categories (BP, 75 cellular components and 171 molecular functions were substantially altered in the TG compared to CG. Many BP were involved in host immune responses (i.e., signaling, signal transmission, signal transduction, response to stimulus, oxidation reduction, response to stress, immune system process, signaling pathway, immune response, cell surface receptor linked signaling pathway. Seven DE gene pathways (VEGF signaling pathway, Long-term potentiation, Ribosome, Asthma, Allograft rejection, Type I diabetes mellitus and Cardiac muscle contraction and statistically significant associations with host responses were affected. Many cytokines (including NRAS, PI3K, MAPK14, CaM, HSP27, protein phosphatase 3, catalytic subunit and alpha isoform, mediating the proliferation and migration of endothelial cells and promoting survival and vascular permeability, were activated in TG, whilst many immunomodulatory cytokines were

  2. Experimental vaccination of pigs with an Actinobacillus pleuropneumoniae serotype 5b capsular polysaccharide tetanus toxoid conjugate

    DEFF Research Database (Denmark)

    Andresen, Lars Ole; Jacobsen, M.J.; Nielsen, J.P.

    1997-01-01

    ) and 8 pigs were vaccinated with Ap5bCP-TT and adjuvant (group 0). Pigs vaccinated with Ap5bCP-TT developed antibody responses to the capsular polysaccharide from A. pleuropneumoniae serotype 5b (Ap5bCP). After challenge, all pigs in groups A and B had severe clinical signs of disease and were euthanized......The protective efficacy of an Actinobacillus pleuropneumoniae serotype 5b capsular polysaccharide-tetanus toroid conjugate (Ap5bCP-TT) against homologous challenge of pigs was investigated. Four pigs were non-vaccinated controls (group A), 4 pigs were injected with adjuvant without antigen (group B...... vaccinated with Ap5bCP-TT had statistically significant reduced values of the mass ratio of affected to unaffected lung tissue compared to pigs in groups A and B (p = 0.01 and p = 0.007, respectively). The results showed that Ap5bCP-TT-vaccination had considerable protective efficacy against lethality...

  3. Effects of Actinobacillus pleuropneumoniae cytotoxins on generation of oxygen radicals by porcine neutrophils

    Directory of Open Access Journals (Sweden)

    Simson Tarigan

    1999-03-01

    Full Text Available Cytotoxins produced by Actinobacillus pleuropneumoniae (App suggested to be the most important pathogenic and virulent factors for this organism. However, the mechanisms on how the cytotoxins contribute to the disease process remain unclear. The purpose of this study is to investigate the effect of the cytotoxins on the oxidative-burst metabolism of porcine neutrophils. In this study, neutrophils were firstly loaded with an oxidative probe dichlorofluorescin diacetate (DCFHDA then expose to cytotoxins. Cells producing oxygen radicals emitted fluorescence and its intensity was measured with a FACScan flow cytometer. All cytotoxins derived from either App serotypes producing ApxI and ApxII, App serotypes producing ApxII only, or App serotypes producing ApxII and ApxIII were capable of stimulating neutrophils for oxygen-radical generation. However, compared with phorbol myristate acetate (PMA, App cytotoxins were much weaker as stimulants for oxygen radicals. In addition, Apx preparation stimulated an oxidative-burst metabolism of neutrophils at a low, narrow range of Apx doses. At higher doses, the toxins inhibit the oxidative burst metabolism. The effects of cytotoxins produced by App during infection on recruited neutrophils into the lungs are assumed to be comparable to those observed in this in vitro study. Neutrophils, and other host cells, adjacent to the bacteria become lysis due to high toxin concentration, whereas those at some distance to the bacteria produce oxygen radicals which in turn cause tissue damage or necrosis.

  4. CO2 Biofixation of Actinobacillus succinogenes Through Novel Amine-Functionalized Polystyrene Microsphere Materials.

    Science.gov (United States)

    Zhu, Wenhao; Li, Qiang; Dai, Ning

    2017-02-01

    CO2-derived succinate production was enhanced by Actinobacillus succinogenes through polystyrene (PSt) microsphere materials for CO2 adsorption in bioreactor, and the adhesion forces between A. succinogenes bacteria and PSt materials were characterized. Synthesized uniformly sized and highly cross-linked PSt microspheres had high specific surface areas. After modification with amine functional groups, the novel amine-functionalized PSt microspheres exhibited a high adsorption capacity of 25.3 mg CO2/g materials. After addition with the functionalized microspheres into the culture broth, CO2 supply to the cells increased. Succinate production by A. succinogenes can be enhanced from 29.6 to 48.1 g L(-1). Moreover, the characterization of interaction forces between A. succinogenes cells and the microspheres indicated that the maximal adhesive force was about 250 pN. The amine-functionalized PSt microspheres can adsorb a large amount of CO2 and be employed for A. succinogenes anaerobic cultivation in bioreactor for high-efficiency production of CO2-derived succinate.

  5. Detection of highly and minimally leukotoxic Actinobacillus actinomycetemcomitans strains in patients with periodontal disease

    Directory of Open Access Journals (Sweden)

    Cortelli Sheila Cavalca

    2003-01-01

    Full Text Available This study examined the prevalence of highly and minimally leukotoxic Actinobacillus actinomycetemcomitans in patients with periodontal disease. Pooled subgingival plaque samples from 136 patients with some form of periodontal disease were examined. Subjects were between 14 and 76 years of age. Clinical examinations included periodontal pocket depth (PD, plaque index (PI and bleeding index (BI. The obtained plaque samples were examined for the presence of highly or minimally leukotoxic A. actinomycetemcomitans strains by the polymerase chain reaction (PCR. Chi-square and logistic regression were performed to evaluate the results. Forty-seven subjects were diagnosed with gingivitis, 70 with chronic periodontitis and 19 with aggressive periodontitis. According to chi-square there was no significant correlation detected between PD (chi2 = 0.73, PI (chi2 = 0.35, BI (chi2 = 0.09 and the presence of the highly leukotoxic A. actinomycetemcomitans. The highly leukotoxic A. actinomycetemcomitans strains were correlated with subjects that were 28 years of age and younger (chi2 = 7.41. There was a significant correlation between highly leukotoxic A. actinomycetemcomitans and aggressive periodontitis (chi2 = 22.06. This study of a Brazilian cohort confirms the strong association between highly leukotoxic A. actinomycetemcomitans strains and the presence of aggressive periodontitis.

  6. Resistance of fluorescent-labelled Actinobacillus actinomycetemcomitans strains to phagocytosis and killing by human neutrophils.

    Science.gov (United States)

    Permpanich, Piyanuj; Kowolik, Michael J; Galli, Dominique M

    2006-01-01

    Neutrophils are initially the predominant cells involved in the host defence of bacterial infections, including periodontal disease. Aggressive periodontitis is associated with Actinobacillus actinomycetemcomitans, a Gram-negative capnophilic microorganism. Infections caused by A. actinomycetemcomitans are not resolved by the host immune response despite the accumulation of neutrophils at the site of inflammation. To better understand the role of natural host defence mechanisms in A. actinomycetemcomitans infections, the interaction of phenotypically diverse strains of this pathogen with human neutrophils was assessed directly using techniques such as genetic labelling with the gene for green fluorescent protein, fluorescence-activated cell sorting and fluorescence imaging. The study included clinical isolates of A. actinomycetemcomitans represented by self-aggregating, biofilm-associated and isogenic planktonic variants. Data obtained showed that complement-mediated phagocytosis of A. actinomycetemcomitans was generally inefficient regardless of strain-specific serotype or leukotoxin production. Furthermore, the majority of ingested bacteria remained viable after exposure to neutrophils for 1 h. Interestingly, uptake of antibody-opsonized bacteria resulted in the rapid cell death of neutrophils. This was in contrast to ingestion of complement-opsonized bacteria, which did not affect neutrophil viability. The methods used in this study provided reliable and reproducible results with respect to adherence, phagocytosis and killing of A. actinomycetemcomitans when encountering human neutrophils.

  7. Killing of Actinobacillus actinomycetemcomitans by the human neutrophil myeloperoxidase-hydrogen peroxide-chloride system.

    Science.gov (United States)

    Miyasaki, K T; Wilson, M E; Genco, R J

    1986-07-01

    Actinobacillus actinomycetemcomitans is a facultative gram-negative coccobacillus associated with periodontal disease and nonoral infections. This organism is resistant to serum bactericidal mechanisms but is nevertheless killed by human neutrophils under aerobic and anaerobic conditions. Most of the killing attributable to oxidative mechanisms is inhibited by sodium cyanide, which suggests that the myeloperoxidase-hydrogen peroxide-chloride (MPO-H2O2-Cl-) system may be a key factor in the oxidative killing process. In this report, we examine whether the isolated MPO-H2O2-Cl- system is bactericidal against A. actinomycetemcomitans. We found that three major chromatographic forms of MPO were capable of killing A. actinomycetemcomitans at sublethal concentrations of H2O2 and that both catalase-positive and catalase-negative strains of this organism were sensitive to killing by the MPO-H2O2-Cl- system. We conclude that the isolated MPO-H2O2-Cl- system is bactericidal for A. actinomycetemcomitans independent of other neutrophil granule constituents and may be an important component of the oxygen-dependent bactericidal activity of the neutrophil with respect to this periodontopathic organism.

  8. Differential killing of Actinobacillus actinomycetemcomitans and Capnocytophaga spp. by human neutrophil granule components.

    Science.gov (United States)

    Miyasaki, K T; Bodeau, A L; Flemmig, T F

    1991-10-01

    The purpose of this study was to determine whether granule fractions of human neutrophils differentially kill Actinobacillus actinomycetemcomitans and Capnocytophaga spp. Granule extracts were subjected to gel filtration, and seven fractions (designated A through G) were obtained. Under aerobic conditions at pH 7.0, representative strains of A. actinomycetemcomitans were killed by fraction D and variably by fraction B. In contrast, the Capnocytophaga spp. were killed by fractions C, D, F, and G. Fractions A (containing lactoferrin and myeloperoxidase) and E (containing lysozyme) exerted little bactericidal activity under these conditions. Anaerobiosis had little effect on the bactericidal activity of fractions D and F but inhibited that of fractions B and C. Electrophoresis, zymography, determination of amino acid composition, and N-terminal sequence analysis revealed that fraction C contained elastase, proteinase 3, and azurocidin. Fraction D contained lysozyme, elastase, and cathepsin G. Subfractions of C and D containing elastase (subfraction C4), a mixture of elastase and azurocidin (subfraction C5), and cathepsin G (subfraction D9) were found to be bactericidal. The bactericidal effects of fraction D and subfraction D9 against A. actinomycetemcomitans was not inhibited by heat inactivation, phenylmethylsulfonyl fluoride, or N-benzyloxycarbonylglycylleucylphenylalanylchloromethyl ketone. We conclude that (i) A. actinomycetemcomitans and Capnocytophaga spp. were sensitive to the bactericidal effects of different neutrophil granule components, (ii) both were sensitive to the bactericidal effects of neutral serine proteases, and (iii) the killing of A. actinomycetemcomitans by cathepsin G-containing fractions was independent of oxygen and neutral serine protease activity.

  9. Succinic Acid Production from Cheese Whey using Actinobacillus succinogenes 130 Z

    Science.gov (United States)

    Wan, Caixia; Li, Yebo; Shahbazi, Abolghasem; Xiu, Shuangning

    Actinobacillus succinogenes 130 Z was used to produce succinic acid from cheese whey in this study. At the presence of external CO2 supply, the effects of initial cheese whey concentration, pH, and inoculum size on the succinic acid production were studied. The by-product formation during the fermentation process was also analyzed. The highest succinic acid yield of 0.57 was obtained at initial cheese whey concentration of 50 g/L, while the highest succinic acid productivity of 0.58 g h-1 L-1 was obtained at initial cheese whey concentration of 100 g/L. Increase in pH and inoculum size caused higher succinic acid yield and productivity. At the preferred fermentation condition of pH 6.8, inoculum size of 5% and initial cheese whey concentration of 50 g/L, succinic acid yield of 0.57, and productivity of 0.44 g h-1 L-1 were obtained. Acetic acid and formic acid were the main by-products throughout the fermentation run of 48 h. It is feasible to produce succinic acid using lactose from cheese whey as carbon resource by A. succinogenes 130 Z.

  10. Apa is a trimeric autotransporter adhesin of Actinobacillus pleuropneumoniae responsible for autoagglutination and host cell adherence.

    Science.gov (United States)

    Xiao, Longwen; Zhou, Liang; Sun, Changjiang; Feng, Xin; Du, ChongTao; Gao, Yu; Ji, Qun; Yang, Shuxin; Wang, Yu; Han, Wenyu; Langford, P R; Lei, Liancheng

    2012-10-01

    Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia, and adherence to host cells is a key step in the pathogenic process. Although trimeric autotransporter adhesins (TAAs) were identified in many pathogenic bacteria in recent years, none in A. pleuropneumoniae have been characterized. In this study, we identified a TAA from A. pleuropneumoniae, Apa, and characterized the contribution of its amino acid residues to the adhesion process. Sequence analysis of the C-terminal amino acid residues of Apa revealed the presence of a putative translocator domain and six conserved HsfBD1-like or HsfBD2-like binding domains. Western blot analysis revealed that the 126 C-terminal amino acids of Apa could form trimeric molecules. By confocal laser scanning microscopy, one of these six domains (ApaBD3) was determined to mediate adherence to epithelial cells. Adherence assays and adherence inhibition assays using a recombinant E. coli- ApaBD3 strain which expressed ApaBD3 on the surface of E. coli confirmed that this domain was responsible for the adhesion activity. Moreover, cellular enzyme-linked immunosorbent assays demonstrated that ApaBD3 mediated high-level adherence to epithelial cell lines. Intriguingly, autoagglutination was observed with the E. coli- ApaBD3 strain, and this phenomenon was dependent upon the association of the expressed ApaBD3 with the C-terminal translocator domain.

  11. Cellular fatty acid composition of Haemophilus species, Pasteurella multocida, Actinobacillus Actinomycetemcomitans and Haemophilus vaginalis (Corynebacterium vaginale).

    Science.gov (United States)

    Jantzen, E; Berdal, B P; Omland, T

    1980-04-01

    The fatty acid composition of 35 Haemophilus influenzae strains was found to be grossly similar and characterized by relatively large amounts of 14:0, 3-OH-14:0, 16:1 and 16:0. The three C18 fatty acids 18:2, 18:1 and 18:0 were also present, but in much lower concentrations. This general pattern was also found for most of the other species of Haemophilus examined (H. aegyptius, H. aphrophilus, H. canis, H. gallinarum, H. haemolyticus, and H. parainfluenzae). Small but distinct quantitative discrepancies were detected for H. ducreyi and the haemin-independent species H. paraphrohaemolyticus, H. paraphrophilus and H. suis. Actinobacillus actinomycetemcomitans was found to be indistinguishable from H. influenzae. Pasteurella multocida also exhibited a fatty acid pattern closely related to that of Haemophilus, but could be distinguished by its higher concentration levels of the C18 fatty acids. The fatty acid pattern of H. vaginalis was considerably different from those of the other species examined. This species lacked 3-OH-14:0 and 18:2 and contained small amounts of 14:0 and 16:0, whereas 18:1 and 18:0 were the major constituents.

  12. Carob pod water extracts as feedstock for succinic acid production by Actinobacillus succinogenes 130Z.

    Science.gov (United States)

    Carvalho, Margarida; Roca, Christophe; Reis, Maria A M

    2014-10-01

    Carob pods are a by-product of locust bean gum industry containing more than 50% (w/w) sucrose, glucose and fructose. In this work, carob pod water extracts were used, for the first time, for succinic acid production by Actinobacillus succinogenes 130Z. Kinetic studies of glucose, fructose and sucrose consumption as individual carbon sources till 30g/L showed no inhibition on cell growth, sugar consumption and SA production rates. Sugar extraction from carob pods was optimized varying solid/liquid ratio and extraction time, maximizing sugar recovery while minimizing the extraction of polyphenols. Batch fermentations containing 10-15g/L total sugars resulted in a maximum specific SA production rate of 0.61Cmol/Cmol X.h, with a yield of 0.55Cmol SA/Cmol sugar and a volumetric productivity of 1.61g SA/L.h. Results demonstrate that carob pods can be a promising low cost feedstock for bio-based SA production.

  13. Improving succinic acid production by Actinobacillus succinogenes from raw industrial carob pods.

    Science.gov (United States)

    Carvalho, Margarida; Roca, Christophe; Reis, Maria A M

    2016-10-01

    Carob pods are an inexpensive by-product of locust bean gum industry that can be used as renewable feedstock for bio-based succinic acid. Here, for the first time, unprocessed raw carob pods were used to extract a highly enriched sugar solution, afterwards used as substrate to produce succinic acid using Actinobacillus succinogenes. Batch fermentations containing 30g/L sugars resulted in a production rate of 1.67gSA/L.h and a yield of 0.39gSA/g sugars. Taking advantage of A. succinogenes' metabolism, uncoupling cell growth from succinic acid production, a fed-batch mode was implemented to increase succinic acid yield and reduce by-products formation. This strategy resulted in a succinic acid yield of 0.94gSA/g sugars, the highest yield reported in the literature for fed-batch and continuous experiments, while maintaining by-products at residual values. Results demonstrate that raw carob pods are a highly efficient feedstock for bio-based succinic acid production.

  14. Optimization of succinic acid fermentation with Actinobacillus succinogenes by response surface methodology (RSM)

    Institute of Scientific and Technical Information of China (English)

    Yun-jian ZHANG; Qiang LI; Yu-xiu ZHANG; Dan WANG; Jian-min XING

    2012-01-01

    Succinic acid is considered as an important platform chemical.Succinic acid fermentation with Actinobacillus succinogenes strain BE-1 was optimized by central composite design (CCD) using a response surface methodology (RSM).The optimized production of succinic acid was predicted and the interactive effects between glucose,yeast extract,and magnesium carbonate were investigated.As a result,a model for predicting the concentration of succinic acid production was developed.The accuracy of the model was confirmed by the analysis of variance (ANOVA),and the validity was further proved by verification experiments showing that percentage errors between actual and predicted values varied from 3.02% to 6.38%.In addition,it was observed that the interactive effect between yeast extract and magnesium carbonate was statistically significant.In conclusion,RSM is an effective and useful method for optimizing the medium components and investigating the interactive effects,and can provide valuable information for succinic acid scale-up fermentation using A.succinogenes strain BE-1.

  15. Molecular characterisation of the early response in pigs to experimental infection with Actinobacillus pleuropneumoniae using cDNA microarrays

    DEFF Research Database (Denmark)

    Hedegaard, Jakob; Skovgaard, Kerstin; Mortensen, Shila;

    2007-01-01

    Background: The bacterium Actinobacillus pleuropneumoniae is responsible for porcine pleuropneumonia, a widespread, highly contagious and often fatal respiratory disease of pigs. The general porcine innate immune response after A. pleuropneumoniae infection is still not clarified. The objective......-infected animals and 130 genes differed in expression in tracheobronchial lymph node tissue from infected versus non-infected animals. Among these genes, several have previously been described to be part of a general host response to infections encoding immune response related proteins. In inflamed lung tissue...

  16. Detection of antibodies to Actinobacillus pleuropneumoniae serotype 12 in pig serum using a blocking enzyme-linked immunosorbent assay

    DEFF Research Database (Denmark)

    Andresen, Lars Ole; Klausen, Joan; Barfod, Kristen

    2002-01-01

    in samples of pig serum were detected by inhibition of the binding of polyclonal rabbit antibodies raised against Ap serotype 12. The assay was evaluated against sera from experimentally infected pigs, from pig herds naturally infected with Ap and from herds declared free of Ap serotypc 12 infection......The objective was to develop a blocking enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to Actinobacillus pleuropneumoniae (Ap) serotype 12 in pig serum. Lipopolysaccharide (LPS) from Ap serotype 12 was purified and used as antigen in the assay. Antibodies to the LPS antigen...

  17. Genomic relationships of Actinobacillus pleuropneumoniae serotype 2 strains evaluated by ribotyping, sequence analysis of ribosomal intergenic regions, and pulsed-field gel electrophoresis

    DEFF Research Database (Denmark)

    Fussing, V.

    1998-01-01

    The aim of the present study was to examine the genomic relationship among 112 Actinobacillus pleuropneumoniae serotype 2 strains obtained throughout Europe and North America. HindIII ribotyping of the strains resulted in five ribotypes of high similarity (87-98%). Sequence analysis of the riboso...

  18. Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to the Apx toxins of Actinobacillus pleuropneumoniae

    DEFF Research Database (Denmark)

    Nielsen, Ragnhild; van den Bosch, Johannes F.; Plambeck, Tamara;

    2000-01-01

    The reference strains of the 12 serotypes of Actinobacillus pleuropneumoniae express one or two of three different RTX exotoxins designated Apr I, Apr II and Apr III. The toxins are important virulence factors. In the present study, ELISAs with purified Apr I, Apr II and Apr III, respectively, as...

  19. Hepatic gene expression changes in pigs experimentally infected with the lung pathogen Actinobacillus pleuropneumoniae as analysed with an innate immunity focused microarray

    DEFF Research Database (Denmark)

    Skovgaard, Kerstin; Mortensen, Shila; Boye, Mette

    2010-01-01

    response of genes associated with innate immune responses was studied in pigs 14–18 h after intranasal inoculation with Actinobacillus pleuropneumoniae, using innate immune focused microarrays and quantitative real-time PCR (qPCR). The microarray analysis of liver tissue established that 51 genes were...

  20. Improved Multiplex PCR Using Conserved and Species-Specific 16S rRNA Gene Primers for Simultaneous Detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis

    OpenAIRE

    Tran, Simon Dangtuan; Rudney, Joel. D.

    1999-01-01

    Among putative periodontal pathogens, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis are most convincingly implicated as etiological agents in periodontitis. Therefore, techniques for detection of those three species would be of value. We previously published a description of a multiplex PCR that detects A. actinomycetemcomitans and P. gingivalis. The present paper presents an improvement on that technique, which now allows more sensitive detection o...

  1. Evaluation of the RapID NH system for identification of Haemophilus somnus, Pasteurella multocida, Pasteurella haemolytica, and Actinobacillus pleuropneumoniae isolated from cattle and pigs with respiratory disease.

    OpenAIRE

    Salmon, S A; Watts, J L; Yancey, R J

    1993-01-01

    Haemophilus somnus, Pasteurella haemolytica, Pasteurella multocida, and Actinobacillus pleuropneumoniae from cattle and pigs with respiratory disease were used to evaluate the RapID NH system (Innovative Diagnostics, Atlanta, Ga.). Minor modifications of the RapID NH system to include animal source and growth requirements would permit the identification of all isolates tested.

  2. A computational strategy for the search of regulatory small RNAs in Actinobacillus pleuropneumoniae

    Science.gov (United States)

    Rossi, Ciro C.; Bossé, Janine T.; Li, Yanwen; Witney, Adam A.; Gould, Kate A.; Langford, Paul R.; Bazzolli, Denise M.S.

    2016-01-01

    Bacterial regulatory small RNAs (sRNAs) play important roles in gene regulation and are frequently connected to the expression of virulence factors in diverse bacteria. Only a few sRNAs have been described for Pasteurellaceae pathogens and no in-depth analysis of sRNAs has been described for Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, responsible for considerable losses in the swine industry. To search for sRNAs in A. pleuropneumoniae, we developed a strategy for the computational analysis of the bacterial genome by using four algorithms with different approaches, followed by experimental validation. The coding strand and expression of 17 out of 23 RNA candidates were confirmed by Northern blotting, RT-PCR, and RNA sequencing. Among them, two are likely riboswitches, three are housekeeping regulatory RNAs, two are the widely studied GcvB and 6S sRNAs, and 10 are putative novel trans-acting sRNAs, never before described for any bacteria. The latter group has several potential mRNA targets, many of which are involved with virulence, stress resistance, or metabolism, and connect the sRNAs in a complex gene regulatory network. The sRNAs identified are well conserved among the Pasteurellaceae that are evolutionarily closer to A. pleuropneumoniae and/or share the same host. Our results show that the combination of newly developed computational programs can be successfully utilized for the discovery of novel sRNAs and indicate an intricate system of gene regulation through sRNAs in A. pleuropneumoniae and in other Pasteurellaceae, thus providing clues for novel aspects of virulence that will be explored in further studies. PMID:27402897

  3. Sub-inhibitory concentrations of penicillin G induce biofilm formation by field isolates of Actinobacillus pleuropneumoniae.

    Science.gov (United States)

    Hathroubi, S; Fontaine-Gosselin, S-È; Tremblay, Y D N; Labrie, J; Jacques, M

    2015-09-30

    Actinobacillus pleuropneumoniae is a Gram-negative bacterium and causative agent of porcine pleuropneumonia. This is a highly contagious disease that causes important economic losses to the swine industry worldwide. Penicillins are extensively used in swine production and these antibiotics are associated with high systemic clearance and low oral bioavailability. This may expose A. pleuropneumoniae to sub-inhibitory concentrations of penicillin G when the antibiotic is administered orally. Our goal was to evaluate the effect of sub-minimum inhibitory concentration (MIC) of penicillin G on the biofilm formation of A. pleuropneumoniae. Biofilm production of 13 field isolates from serotypes 1, 5a, 7 and 15 was tested in the presence of sub-MIC of penicillin G using a polystyrene microtiter plate assay. Using microscopy techniques and enzymatic digestion, biofilm architecture and composition were also characterized after exposure to sub-MIC of penicillin G. Sub-MIC of penicillin G significantly induced biofilm formation of nine isolates. The penicillin G-induced biofilms contained more poly-N-acetyl-D-glucosamine (PGA), extracellular DNA and proteins when compared to control biofilms grown without penicillin G. Additionally, penicillin G-induced biofilms were sensitive to DNase which was not observed with the untreated controls. Furthermore, sub-MIC of penicillin G up-regulated the expression of pgaA, which encodes a protein involved in PGA synthesis, and the genes encoding the envelope-stress sensing two-component regulatory system CpxRA. In conclusion, sub-MICs of penicillin G significantly induce biofilm formation and this is likely the result of a cell envelope stress sensed by the CpxRA system resulting in an increased production of PGA and other matrix components.

  4. Absence of TolC Impairs Biofilm Formation in Actinobacillus pleuropneumoniae by Reducing Initial Attachment

    Science.gov (United States)

    Yuan, Jianlin; Lau, Gee W.; Wen, Yiping; Wu, Rui; Zhao, Qin; Huang, Xiaobo; Yan, Qigui; Huang, Yong; Wen, Xintian

    2016-01-01

    Actinobacillus pleuropneumoniae is the etiologic agent of porcine contagious pleuropneumonia, a major cause of economic loss in swine industry worldwide. TolC, the key component of multidrug efflux pumps and type I secretion systems, has been well-studied as an exit duct for numerous substances in many Gram-negative bacteria. By contrast, little is known on the role of TolC in biofilm formation. In this study, a ΔtolC mutant was used to examine the importance of TolC in biofilm formation of A. pleuropneumoniae. Surface attachment assays demonstrated the essential role of TolC in initial attachment of biofilm cells. The loss of TolC function altered surface hydrophobicity, and resulted in greatly reduced autoaggregation in ΔtolC. Using both enzymatic treatments and confocal microscopy, biofilm composition and architecture were characterized. When compared against the wild-type strain, the poly-β-1, 6-N-acetyl-D-glucosamine (PGA), an important biofilm matrix component of A. pleuropneumoniae, was significantly reduced at the initial attachment stage in ΔtolC. These results were confirmed by mRNA level using quantitative RT-PCR. Additionally, defective secretion systems in ΔtolC may also contribute to the deficiency in biofilm formation. Taken together, the current study demonstrated the importance of TolC in the initial biofilm formation stage in A. pleuropneumoniae. These findings could have important clinical implications in developing new treatments against biofilm-related infections by A. pleuropneumoniae. PMID:27681876

  5. Whole Genome Sequencing for Surveillance of Antimicrobial Resistance in Actinobacillus pleuropneumoniae

    Science.gov (United States)

    Bossé, Janine T.; Li, Yanwen; Rogers, Jon; Fernandez Crespo, Roberto; Li, Yinghui; Chaudhuri, Roy R.; Holden, Matthew T. G.; Maskell, Duncan J.; Tucker, Alexander W.; Wren, Brendan W.; Rycroft, Andrew N.; Langford, Paul R.

    2017-01-01

    The aim of this study was to evaluate the correlation between antimicrobial resistance (AMR) profiles of 96 clinical isolates of Actinobacillus pleuropneumoniae, an important porcine respiratory pathogen, and the identification of AMR genes in whole genome sequence (wgs) data. Susceptibility of the isolates to nine antimicrobial agents (ampicillin, enrofloxacin, erythromycin, florfenicol, sulfisoxazole, tetracycline, tilmicosin, trimethoprim, and tylosin) was determined by agar dilution susceptibility test. Except for the macrolides tested, elevated MICs were highly correlated to the presence of AMR genes identified in wgs data using ResFinder or BLASTn. Of the isolates tested, 57% were resistant to tetracycline [MIC ≥ 4 mg/L; 94.8% with either tet(B) or tet(H)]; 48% to sulfisoxazole (MIC ≥ 256 mg/L or DD = 6; 100% with sul2), 20% to ampicillin (MIC ≥ 4 mg/L; 100% with blaROB-1), 17% to trimethoprim (MIC ≥ 32 mg/L; 100% with dfrA14), and 6% to enrofloxacin (MIC ≥ 0.25 mg/L; 100% with GyrAS83F). Only 33% of the isolates did not have detectable AMR genes, and were sensitive by MICs for the antimicrobial agents tested. Although 23 isolates had MIC ≥ 32 mg/L for tylosin, all isolates had MIC ≤ 16 mg/L for both erythromycin and tilmicosin, and no macrolide resistance genes or known point mutations were detected. Other than the GyrAS83F mutation, the AMR genes detected were mapped to potential plasmids. In addition to presence on plasmid(s), the tet(B) gene was also found chromosomally either as part of a 56 kb integrative conjugative element (ICEApl1) in 21, or as part of a Tn7 insertion in 15 isolates. Our results indicate that, with the exception of macrolides, wgs data can be used to accurately predict resistance of A. pleuropneumoniae to the tested antimicrobial agents and provides added value for routine surveillance.

  6. Actinobacillus pleuropneumoniae possesses an antiviral activity against porcine reproductive and respiratory syndrome virus.

    Directory of Open Access Journals (Sweden)

    Cynthia Lévesque

    Full Text Available Pigs are often colonized by more than one bacterial and/or viral species during respiratory tract infections. This phenomenon is known as the porcine respiratory disease complex (PRDC. Actinobacillus pleuropneumoniae (App and porcine reproductive and respiratory syndrome virus (PRRSV are pathogens that are frequently involved in PRDC. The main objective of this project was to study the in vitro interactions between these two pathogens and the host cells in the context of mixed infections. To fulfill this objective, PRRSV permissive cell lines such as MARC-145, SJPL, and porcine alveolar macrophages (PAM were used. A pre-infection with PRRSV was performed at 0.5 multiplicity of infection (MOI followed by an infection with App at 10 MOI. Bacterial adherence and cell death were compared. Results showed that PRRSV pre-infection did not affect bacterial adherence to the cells. PRRSV and App co-infection produced an additive cytotoxicity effect. Interestingly, a pre-infection of SJPL and PAM cells with App blocked completely PRRSV infection. Incubation of SJPL and PAM cells with an App cell-free culture supernatant is also sufficient to significantly block PRRSV infection. This antiviral activity is not due to LPS but rather by small molecular weight, heat-resistant App metabolites (<1 kDa. The antiviral activity was also observed in SJPL cells infected with swine influenza virus but to a much lower extent compared to PRRSV. More importantly, the PRRSV antiviral activity of App was also seen with PAM, the cells targeted by the virus in vivo during infection in pigs. The antiviral activity might be due, at least in part, to the production of interferon γ. The use of in vitro experimental models to study viral and bacterial co-infections will lead to a better understanding of the interactions between pathogens and their host cells, and could allow the development of novel prophylactic and therapeutic tools.

  7. Growth of Actinobacillus pleuropneumoniae is promoted by exogenous hydroxamate and catechol siderophores.

    Science.gov (United States)

    Diarra, M S; Dolence, J A; Dolence, E K; Darwish, I; Miller, M J; Malouin, F; Jacques, M

    1996-03-01

    Siderophores bind ferric ions and are involved in receptor-specific iron transport into bacteria. Six types of siderophores were tested against strains representing the 12 different serotypes of Actinobacillus pleuropneumoniae. Ferrichrome and bis-catechol-based siderophores showed strong growth-promoting activities for A. pleuropneumoniae in a disk diffusion assay. Most strains of A. pleuropneumoniae tested were able to use ferrichrome (21 of 22 or 95%), ferrichrome A (20 of 22 or 90%), and lysine-based bis-catechol (20 of 22 or 90%), while growth of 36% (8 of 22) was promoted by a synthetic hydroxamate, N5-acetyl-N5-hydroxy-L-ornithine tripeptide. A. pleuropneumoniae serotype 1 (strain FMV 87-682) and serotype 5 (strain 2245) exhibited a distinct yellow halo around colonies on Chrome Azurol S agar plates, suggesting that both strains can produce an iron chelator (siderophore) in response to iron stress. The siderophore was found to be neither a phenolate nor a hydroxamate by the chemical tests of Arnow and Csaky, respectively. This is the first report demonstrating the production of an iron chelator and the use of exogenous siderophores by A. pleuropneumoniae. A spermidine-based bis-catechol siderophore conjugated to a carbacephalosporin was shown to inhibit growth of A. pleuropneumoniae. A siderophore-antibiotic-resistant strain was isolated and shown to have lost the ability to use ferrichrome, synthetic hydroxamate, or catechol-based siderophores when grown under conditions of iron restriction. This observation indicated that a common iron uptake pathway, or a common intermediate, for hydroxamate- and catechol-based siderophores may exist in A. pleuropneumoniae.

  8. Role of (p)ppGpp in Viability and Biofilm Formation of Actinobacillus pleuropneumoniae S8.

    Science.gov (United States)

    Li, Gang; Xie, Fang; Zhang, Yanhe; Bossé, Janine T; Langford, Paul R; Wang, Chunlai

    2015-01-01

    Actinobacillus pleuropneumoniae is a Gram-negative bacterium and the cause of porcine pleuropneumonia. When the bacterium encounters nutritional starvation, the relA-dependent (p)ppGpp-mediated stringent response is activated. The modified nucleotides guanosine 5'-diphosphate 3'-diphosphate (ppGpp) and guanosine 5'-triphosphate 3'-diphosphate (pppGpp) are known to be signaling molecules in other prokaryotes. Here, to investigate the role of (p)ppGpp in A. pleuropneumoniae, we created a mutant A. pleuropneumoniae strain, S8ΔrelA, which lacks the (p)ppGpp-synthesizing enzyme RelA, and investigated its phenotype in vitro. S8ΔrelA did not survive after stationary phase (starvation condition) and grew exclusively as non-extended cells. Compared to the wild-type (WT) strain, the S8ΔrelA mutant had an increased ability to form a biofilm. Transcriptional profiles of early stationary phase cultures revealed that a total of 405 bacterial genes were differentially expressed (including 380 up-regulated and 25 down-regulated genes) in S8ΔrelA as compared with the WT strain. Most of the up-regulated genes are involved in ribosomal structure and biogenesis, amino acid transport and metabolism, translation cell wall/membrane/envelope biogenesis. The data indicate that (p)ppGpp coordinates the growth, viability, morphology, biofilm formation and metabolic ability of A. pleuropneumoniae in starvation conditions. Furthermore, S8ΔrelA could not use certain sugars nor produce urease which has been associated with the virulence of A. pleuropneumoniae, suggesting that (p)ppGpp may directly or indirectly affect the pathogenesis of A. pleuropneumoniae during the infection process. In summary, (p)ppGpp signaling represents an essential component of the regulatory network governing stress adaptation and virulence in A. pleuropneumoniae.

  9. Differential gene expression profiling of Actinobacillus pleuropneumoniae during induction of primary alveolar macrophage apoptosis in piglets.

    Science.gov (United States)

    Wang, Lei; Qin, Wanhai; Ruidong, Zhai; Liu, Shiting; Zhang, Hu; Sun, Changjiang; Feng, Xin; Gu, Jingmin; Du, Chongtao; Han, Wenyu; Langford, P R; Lei, Liancheng

    2015-01-01

    Actinobacillus pleuropneumoniae (A. pleuropneumoniae) is the causative agent of porcine pleuropneumonia, a disease that causes serious problems for the swine industry. Successful infection by this bacterium requires breaking the first line of defence in the lungs, the primary alveolar macrophages (PAMs). Therefore, exploring A. pleuropneumoniae-PAM interactions will provide vital groundwork for the scientific control of this infectious disease, which has been little studied up to now. In this work, PAMs were isolated from piglets and co-incubated with A. pleuropneumoniae serovar 5b strain L20 in vitro, and their interaction, PAM cell death, and differential gene expression of A. pleuropneumoniae in response to PAM cell death were observed and analysed using confocal microscopy, electron microscopy, RT-PCR, Western blot, flow cytometry and the use of a gene expression profile chip. A. pleuropneumoniae quickly adhered to and invaded PAMs, inducing apoptosis, which was confirmed using transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The highest percentage of apoptosis in cells was confirmed using flow cytometry when the cells were infected at a multiplicity of infection (MOI) of 10 and incubated for 5 h, with higher expression of activated caspase-3 as measured by Western blot. Using microarray gene chips with 2868 probes containing nearly all of the genomic sequence of A. pleuropneumoniae serotype 5b strain L20, a total of 185 bacterial genes were found to be differentially expressed (including 92 up-regulated and 93 down-regulated genes) and involved in the process of apoptosis, as compared with the expression of control bacteria cultured without PAMs in BHI medium (mean expression ratios >1.5-fold, p PAMs and undergoes complex changes in gene transcription, including expression changes in known and potential virulence factors. Some potentially novel virulence targets have been identified, suggesting new strategies for the

  10. In vitro activity of antibiotics alone and in combination against Actinobacillus actinomycetemcomitans.

    Science.gov (United States)

    Yogev, R; Shulman, D; Shulman, S T; Glogowski, W G

    1986-01-01

    The MICs for 90% of the organisms tested (MIC90S) of 11 antibiotics against 24 clinical isolates of Actinobacillus actinomycetemcomitans were determined by the MIC 2000 system. The lowest MIC90S (16 micrograms/ml) were observed with ceftriaxone and rifampin. The next lowest MIC90S were found with cephapirin, tetracycline, and chloramphenicol (3.12 micrograms/ml). The MIC90S of penicillin, ampicillin, ticarcillin, piperacillin, and amikacin were each greater than or equal to 12.5 micrograms/ml. Antibiotic synergy was studied by the killing curve method and was defined as a greater than or equal to 2 log10 reduction in CFU when two antibiotics were used in combination at one-fourth the MBC for each compared with the effect of each antibiotic alone at one-half the MBC. Synergism between rifampin and penicillin, cephapirin, or ceftriaxone was tested for with 12 A. actinomycetemcomitans strains. In 7 of 37 instances, synergism was demonstrated for the combinations rifampin plus ceftriaxone (n = 3) or rifampin plus penicillin (n = 4); in 9 instances, an additive effect was noted, and impaired killing with drug combinations compared with the effect of a single antibiotic was suggested in 4 strains. The majority of strains were indifferent to the combinations. Similarly, variable results were observed when the combination of trimethoprim and cephapirin was tested against eight A. actinomycetemcomitans strains. Our data suggest that rifampin and cephapirin are the most active of the 11 antibiotics studied against A. actinomycetemcomitans. In addition, in vitro synergism between rifampin and other antibiotics or between trimethoprim and cephapirin was not consistently demonstrable.

  11. Role of (pppGpp in Viability and Biofilm Formation of Actinobacillus pleuropneumoniae S8.

    Directory of Open Access Journals (Sweden)

    Gang Li

    Full Text Available Actinobacillus pleuropneumoniae is a Gram-negative bacterium and the cause of porcine pleuropneumonia. When the bacterium encounters nutritional starvation, the relA-dependent (pppGpp-mediated stringent response is activated. The modified nucleotides guanosine 5'-diphosphate 3'-diphosphate (ppGpp and guanosine 5'-triphosphate 3'-diphosphate (pppGpp are known to be signaling molecules in other prokaryotes. Here, to investigate the role of (pppGpp in A. pleuropneumoniae, we created a mutant A. pleuropneumoniae strain, S8ΔrelA, which lacks the (pppGpp-synthesizing enzyme RelA, and investigated its phenotype in vitro. S8ΔrelA did not survive after stationary phase (starvation condition and grew exclusively as non-extended cells. Compared to the wild-type (WT strain, the S8ΔrelA mutant had an increased ability to form a biofilm. Transcriptional profiles of early stationary phase cultures revealed that a total of 405 bacterial genes were differentially expressed (including 380 up-regulated and 25 down-regulated genes in S8ΔrelA as compared with the WT strain. Most of the up-regulated genes are involved in ribosomal structure and biogenesis, amino acid transport and metabolism, translation cell wall/membrane/envelope biogenesis. The data indicate that (pppGpp coordinates the growth, viability, morphology, biofilm formation and metabolic ability of A. pleuropneumoniae in starvation conditions. Furthermore, S8ΔrelA could not use certain sugars nor produce urease which has been associated with the virulence of A. pleuropneumoniae, suggesting that (pppGpp may directly or indirectly affect the pathogenesis of A. pleuropneumoniae during the infection process. In summary, (pppGpp signaling represents an essential component of the regulatory network governing stress adaptation and virulence in A. pleuropneumoniae.

  12. Improved diagnostic PCR assay for Actinobacillus pleuropneumoniae based on the nucleotide sequence of an outer membrane lipoprotein

    DEFF Research Database (Denmark)

    Gram, Trine; Ahrens, Peter

    1998-01-01

    The gene (omlA) coding for an outer membrane protein of Actinobacillus pleuropneumoniae serotypes 1 and 5 has been described earlier and has formed the basis for development of a specific PCR assay, The corresponding regions of all 12 A. pleuropneumoniae reference strains of biovar 1 were sequenc...... and sensitivity of this PCR compared to those of culture suggest the use of this PCR for routine identification of A. pleuropneumoniae.......The gene (omlA) coding for an outer membrane protein of Actinobacillus pleuropneumoniae serotypes 1 and 5 has been described earlier and has formed the basis for development of a specific PCR assay, The corresponding regions of all 12 A. pleuropneumoniae reference strains of biovar 1 were sequenced...... species related to A. pleuropneumoniae or isolated from pigs were assayed. They were all found negative in the PCR, as were tonsil cultures from 50 pigs of an A. pleuropneumoniae-negative herd. The sensitivity assessed by agarose gel analysis of the PCR product was 10(2) CFU/PCR test tube. The specificity...

  13. An indirect enzyme-linked immunosorbent assay for detection of antibodies to Actinobacillus pleuropneumoniae serovar 7 in pig serum

    DEFF Research Database (Denmark)

    Klausen, Joan; Ekeroth, Lars; Grondahl-Hansen, Jan;

    2007-01-01

    Lipopolysaccharide (LPS) antigen was purified from Actinobacillus pleuropneumoniae serovar 7 by phenol-water extraction and fractionated on a, S-100 Sephacryl column. High molecular weight fractions of LPS purified from the S-100 column were pooled and used as antigen in an indirect serovar 7 ELI...... as well as sera from herds free of infection with A. pleuropneumoniae serovar 7. When compared to the complement fixation test (CFT) as a reference test, the ELISA showed much higher sensitivity and statistically equivalent specificity.......Lipopolysaccharide (LPS) antigen was purified from Actinobacillus pleuropneumoniae serovar 7 by phenol-water extraction and fractionated on a, S-100 Sephacryl column. High molecular weight fractions of LPS purified from the S-100 column were pooled and used as antigen in an indirect serovar 7 ELISA....... The ELISA was evaluated with sera from pigs experimentally infected with 11 different A. pleuropneumoniae serovars of biotype 1. Estimation of sensitivity and specificity of the A. pleuropneumoniae serovar 7 ELISA was performed using pig sera from herds naturally infected with A. pleuropneumoniae serovar 7...

  14. Characterization of Biofilm Formation in [Pasteurella] pneumotropica and [Actinobacillus] muris Isolates of Mouse Origin.

    Science.gov (United States)

    Sager, Martin; Benten, W Peter M; Engelhardt, Eva; Gougoula, Christina; Benga, Laurentiu

    2015-01-01

    [Pasteurella] pneumotropica biotypes Jawetz and Heyl and [Actinobacillus] muris are the most prevalent Pasteurellaceae species isolated from laboratory mouse. However, mechanisms contributing to their high prevalence such as the ability to form biofilms have not been studied yet. In the present investigation we analyze if these bacterial species can produce biofilms in vitro and investigate whether proteins, extracellular DNA and polysaccharides are involved in the biofilm formation and structure by inhibition and dispersal assays using proteinase K, DNase I and sodium periodate. Finally, the capacity of the biofilms to confer resistance to antibiotics is examined. We demonstrate that both [P.] pneumotropica biotypes but not [A.] muris are able to form robust biofilms in vitro, a phenotype which is widely spread among the field isolates. The biofilm inhibition and dispersal assays by proteinase and DNase lead to a strong inhibition in biofilm formation when added at the initiation of the biofilm formation and dispersed pre-formed [P.] pneumotropica biofilms, revealing thus that proteins and extracellular DNA are essential in biofilm formation and structure. Sodium periodate inhibited the bacterial growth when added at the beginning of the biofilm formation assay, making difficult the assessment of the role of β-1,6-linked polysaccharides in the biofilm formation, and had a biofilm stimulating effect when added on pre-established mature biofilms of [P.] pneumotropica biotype Heyl and a majority of [P.] pneumotropica biotype Jawetz strains, suggesting that the presence of β-1,6-linked polysaccharides on the bacterial surface might attenuate the biofilm production. Conversely, no effect or a decrease in the biofilm quantity was observed by biofilm dispersal using sodium periodate on further biotype Jawetz isolates, suggesting that polysaccharides might be incorporated in the biofilm structure. We additionally show that [P.] pneumotropica cells enclosed in biofilms

  15. Actinobacillus actinomycetemcomitans and localized juvenile periodontitis. Clinical, microbiologic and histologic studies.

    Science.gov (United States)

    Christersson, L A

    1993-01-01

    The present studies examined Actinobacillus actinomycetemcomitans and its role in localized juvenile periodontitis (LJP). The distribution of the bacteria was studied in healthy normals, patients with adult periodontitis, diabetics, and those with LJP. Over 95% of the LJP patients harbored A. actinomycetemcomitans, whereas only 17% of healthy subjects, 21% of adult periodontitis patients, and 5% of diabetics were positive. All members of a LJP family harboring the organism yielded isolates of the same biotype and serotype. The transmission of the bacteria was studied after transfer of the bacteria, with periodontal probes from infected to healthy gingival sites, within the oral cavity of LJP patients. Newly colonized gingival sites, 50% of those involved, became free of A. actinomycetemcomitans after only 3 weeks. A purposely forceful inoculation contributed to a more predictable colonization (89%), but only prolonged the colonization with one week. Treatment of LJP lesions with scaling and root planing resulted in minimal clinical and microbiological changes during a 16 week follow-up period. However, gingival curettage and modified Widman flap surgery suppressed A. actinomycetemcomitans in 75% and 89% of the sites, and resulted in resolution of periodontal pocket depth and gain in attachment level. Gingival tissue specimens, from 35 LJP sites, 3 control sites, and one monkey biopsy, were studied to verify the hypothesis of gingival infiltration of A. actinomycetemcomitans. Bacteria were identified immunohistologically with rabbit antisera serospecific to the three A. actinomycetemcomitans serotypes. Positive staining was observed in the tissue from all but one LJP patient. Twenty-eight (80%) lesions were positive for A. actinomycetemcomitans antigens in the gingival connective tissue, often with antigens located both between and within cells. The specimen from a culture positive control demonstrated no signs of invasion, similar to the monkey specimen

  16. [Breeding of Actinobacillus succiniogenes mutants with improved succinate production based on metabolic flux analysis].

    Science.gov (United States)

    Pan, Lijun; Li, Xingjiang; Jiang, Shaotong; Wei, Zhaojun; Chen, Xiaohui; Cai, Licheng; Wang, Hefeng; Jiang, Jijun

    2008-09-01

    It is very important to obtain high yield mutant strains on the base of metabolic flux analysis of Actinobacillus succinogenes S.JST for the industrial bioconversion of succinic acid. The metabolic pathway was analized at first and the flux of the metabolic networks was calculated by matrix. In order to decrease acetic acid flux, the strains mutated by soft X-ray of synchronous radiation were screened on the plates with high concentration of fluoroacetic acid. For decreasing the metabolic flux of ethanol the site-directed mutagenesis was carried out for the reduction of alcohol dehydrogenase(Adh) specific activity. Then the enzyme activity determination and the gene sequence analysis of the mutant strain was compared with those of the parent strain. Metabolic flux analysis of the parent strain indicated that the flux of succinic acid was 1.78(mmol/g/h) and that the flux of acetic acid and ethanol were 0.60 (mmol/g/h) and 1.04( mmol/g/h), respectively. Meanwhile the metabolic pathway analysis showed that the ethanol metabolism enhanced the lacking of H electron donor during the synthesis of succinic acid and that the succinic acid flux was weakened by the metabolism of byproducts ethanol and acetic acid. Compared with the parent strain, the acetic acid flux of anti-fluoroacetic mutant strain S.JST1 was 0.024 (mmol/g/h), decreasing by 96%. Then the enzyme determination showed that the specific activity unit of phosphotransacetylase(Pta) decreased from 602 to 74 and a mutated site was founded in the pta gene of the mutant strain S.JST1. Compared with that of the parent strain S.JST1 the ethanol flux of adh-site-directed mutant strain S.JST2 was 0.020 (mmol/g/h), decreasing by 98%. Then the enzyme determination showed that the specific activity unit of Adh decreased from 585 to 62 and the yield of end product succinic acid was 65.7 (g/L). The interdiction of Adh and Pta decreased the metabolism of byproducts and the H electron donor was well balanced, thus the succinic

  17. The role of Actinobacillus actinomycetemcomitans fimbrial adhesin on MMP-8 activity in aggressive periodontitis pathogenesis

    Directory of Open Access Journals (Sweden)

    Rini Devijanti Ridwan

    2012-12-01

    Full Text Available Background: Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans is Gram negative and a major bacterial agent associated with aggressive periodontitis in young adult, this bacteria was an important factor in pathogenesis of aggressive periodontitis. A. actinomycetemcomitans possesses fimbriae with an adhesin protein that was the first bacterial molecules to make physical contact with host. Purpose: The objective of this research was to analyzed the influence of A. actinomycetemcomitans fimbrial adhesin protein induction on MMP-8 activity. Methods: The research was an experimental laboratory study, the step in this study were isolation and identification A. actinomycetemcomitans, characterize A. actinomycetemcomitans adhesin and study the role of A. actinomycetemcomitans adhesin in Wistar rats. Results: The result of this research on the role of adhesin in Wistar rats after analysis with Analysis of Variance (ANOVA showed significant differences in the control group with group induction with A. actinomycetemcomitans, A. actinomycetemcomitans plus adhesin and adhesin. MMP-8 activity increased with induction A. actinomycetemcomitans and 24 kDa A. actinomycetemcomitans adhesin. This fimbrial adhesin protein showed that A. actinomycetemcomitans has the ability to adhesion, colonization and invasion for host in aggressive periodontitis pathogenesis. Conclusion: A. actinomycetemcomitans fimbrial adhesin protein induction increasing MMP-8 activity for aggressive periodontitis pathogenesis.Latar belakang: A. actinomycetemcomitans merupakan salah satu bakteri Gram negatif yang terkait dengan periodontitis agresif yang menyerang penderita usia muda dan merupakan faktor penting dalam patogenesis periodontitis agresif. A. actimycetemcomitans mempunyai fimbriae dengan protein adhesin yang merupakan molekul pertama dari bakteri untuk melakukan kontak fisik dengan host. Tujuan: Tujuan penelitian ini adalah menganalisis pengaruh induksi adhesin A

  18. Transcriptional profiling of Actinobacillus pleuropneumoniae during the acute phase of a natural infection in pigs

    Directory of Open Access Journals (Sweden)

    Harel Josée

    2010-02-01

    Full Text Available Abstract Background Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, a respiratory disease which causes great economic losses worldwide. Many virulence factors are involved in the pathogenesis, namely capsular polysaccharides, RTX toxins, LPS and many iron acquisition systems. In order to identify genes that are expressed in vivo during a natural infection, we undertook transcript profiling experiments with an A. pleuropneumoniae DNA microarray, after recovery of bacterial mRNAs from serotype 5b-infected porcine lungs. AppChip2 contains 2033 PCR amplicons based on the genomic sequence of App serotype 5b strain L20, representing more than 95% of ORFs greater than 160 bp in length. Results Transcriptional profiling of A. pleuropneumoniae recovered from the lung of a pig suffering from a natural infection or following growth of the bacterial isolate in BHI medium was performed. An RNA extraction protocol combining beadbeating and hot-acid-phenol was developed in order to maximize bacterial mRNA yields and quality following total RNA extraction from lung lesions. Nearly all A. pleuropneumoniae transcripts could be detected on our microarrays, and 150 genes were deemed differentially expressed in vivo during the acute phase of the infection. Our results indicate that, for example, gene apxIVA from an operon coding for RTX toxin ApxIV is highly up-regulated in vivo, and that two genes from the operon coding for type IV fimbriae (APL_0878 and APL_0879 were also up-regulated. These transcriptional profiling data, combined with previous comparative genomic hybridizations performed by our group, revealed that 66 out of the 72 up-regulated genes are conserved amongst all serotypes and that 3 of them code for products that are predicted outer membrane proteins (genes irp and APL_0959, predicted to code for a TonB-dependent receptor and a filamentous hemagglutinin/adhesin respectively or lipoproteins (gene APL_0920. Only 4

  19. Site-specific subgingival colonization by Actinobacillus actinomycetemcomitans in orthodontic patients.

    Science.gov (United States)

    Paolantonio, M; Festa, F; di Placido, G; D'Attilio, M; Catamo, G; Piccolomini, R

    1999-04-01

    A high prevalence of Actinobacillus actinomycetemcomitans (Aa) in subgingival plaque in patients for orthodontia already has been observed. The present study had the following aims: 1) to ascertain a direct relationship between the orthodontic appliance placement and the subgingival colonization by Aa, and 2) to determine whether the Aa growth specifically occurred on teeth with braces attached or whether the presence of orthodontic appliances could also cause the isolation of Aa in teeth free from therapeutic appliances. Twenty-four young systemically and periodontally healthy subjects with malaligned and crowded teeth in the anterior sextants of both dental arches participated in this study. After 1 session of ultrasonic scaling with oral hygiene instructions during the first experimental session, the mesiobuccal sites of the first molars and the distobuccal sites of the lateral incisors in both dental arches in each participant were subjected to clinical and microbiologic examination for the recovery of Aa. Clinical examination consisted of recording the presence of plaque and the examination of gingival bleeding on probing and probing depth. Microbiologic sampling was obtained with the insertion of 3 sterile paper points at the deepest part of each gingival sulcus. Altogether, 192 periodontal sites were examined. After the examinations, the patients received fixed orthodontic appliances in only 1 dental arch (test sites) and the other one was left free from appliances (control sites). Clinical examination and microbiologic sampling were repeated in the same experimental test and control sites after 4, 8, and 12 weeks. At the 12-week session, the orthodontic appliance was removed from the test arch, and, 4 weeks later, a further clinical and microbiologic examination was performed. The results showed that, during the period with orthodontic appliances, the presence of plaque scores and the gingival bleeding on probing scores were increased significantly and that

  20. Proteomic and immunoproteomic characterization of a DIVA subunit vaccine against Actinobacillus pleuropneumoniae

    Directory of Open Access Journals (Sweden)

    Maas Alexander

    2011-04-01

    Full Text Available Abstract Background Protection of pigs by vaccination against Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is hampered by the presence of 15 different serotypes. A DIVA subunit vaccine comprised of detergent-released proteins from A. pleuropneumoniae serotypes 1, 2 and 5 has been developed and shown to protect pigs from clinical symptoms upon homologous and heterologous challenge. This vaccine has not been characterized in-depth so far. Thus we performed i mass spectrometry in order to identify the exact protein content of the vaccine and ii cross-serotype 2-D immunoblotting in order to discover cross-reactive antigens. By these approaches we expected to gain results enabling us to argue about the reasons for the efficacy of the analyzed vaccine. Results We identified 75 different proteins in the vaccine. Using the PSORTb algorithm these proteins were classified according to their cellular localization. Highly enriched proteins are outer membrane-associated lipoproteins like OmlA and TbpB, integral outer membrane proteins like FrpB, TbpA, OmpA1, OmpA2, HgbA and OmpP2, and secreted Apx toxins. The subunit vaccine also contained large amounts of the ApxIVA toxin so far thought to be expressed only during infection. Applying two-dimensional difference gel electrophoresis (2-D DIGE we showed different isoforms and variations in expression levels of several proteins among the strains used for vaccine production. For detection of cross-reactive antigens we used detergent released proteins of serotype 7. Sera of pigs vaccinated with the detergent-released proteins of serotypes 1, 2, and 5 detected seven different proteins of serotype 7, and convalescent sera of pigs surviving experimental infection with serotype 7 reacted with 13 different proteins of the detergent-released proteins of A. pleuropneumoniae serotypes 1, 2, and 5. Conclusions A detergent extraction-based subunit vaccine of A. pleuropneumoniae was

  1. Microarray-based comparative genomic profiling of reference strains and selected Canadian field isolates of Actinobacillus pleuropneumoniae

    Directory of Open Access Journals (Sweden)

    MacInnes Janet I

    2009-02-01

    Full Text Available Abstract Background Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is a highly contagious respiratory pathogen that causes severe losses to the swine industry worldwide. Current commercially-available vaccines are of limited value because they do not induce cross-serovar immunity and do not prevent development of the carrier state. Microarray-based comparative genomic hybridizations (M-CGH were used to estimate whole genomic diversity of representative Actinobacillus pleuropneumoniae strains. Our goal was to identify conserved genes, especially those predicted to encode outer membrane proteins and lipoproteins because of their potential for the development of more effective vaccines. Results Using hierarchical clustering, our M-CGH results showed that the majority of the genes in the genome of the serovar 5 A. pleuropneumoniae L20 strain were conserved in the reference strains of all 15 serovars and in representative field isolates. Fifty-eight conserved genes predicted to encode for outer membrane proteins or lipoproteins were identified. As well, there were several clusters of diverged or absent genes including those associated with capsule biosynthesis, toxin production as well as genes typically associated with mobile elements. Conclusion Although A. pleuropneumoniae strains are essentially clonal, M-CGH analysis of the reference strains of the fifteen serovars and representative field isolates revealed several classes of genes that were divergent or absent. Not surprisingly, these included genes associated with capsule biosynthesis as the capsule is associated with sero-specificity. Several of the conserved genes were identified as candidates for vaccine development, and we conclude that M-CGH is a valuable tool for reverse vaccinology.

  2. Multiplex PCR using conserved and species-specific 16S rRNA gene primers for simultaneous detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis.

    OpenAIRE

    Tran, S D; Rudney, J. D.

    1996-01-01

    Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis are strongly associated with periodontitis. However, little is known about their distribution in periodontally healthy individuals, because culturing techniques are not sufficiently sensitive. A modified multiplex PCR was developed to address that question. Our method uses two species-specific forward primers in combination with a single reverse primer. These primers target variable and conserved regions of the 16S rRNA gene. S...

  3. Serotype-related differences in production and type of heat-labile hemolysin and heat-labile cytotoxin of Actinobacillus (Haemophilus) pleuropneumoniae.

    OpenAIRE

    Kamp, E M; van Leengoed, L A

    1989-01-01

    Reference strains of serotypes 1 to 12 of Actinobacillus (Haemophilus) pleuropneumoniae were cultured in Eagle minimal essential medium with 10% Serum Plus. Culture supernatants were examined for cytotoxicity to alveolar macrophages and for the ability to hemolyze sheep erythrocytes. All strains except the reference strain of serotype 6 produced cytotoxin, whereas only serotypes 1, 5, 9, 10, and 11 produced hemolysin. Both cytotoxin and hemolysin appeared to be heat labile. Antisera were rais...

  4. Molecular characterisation of the early response in pigs to experimental infection with Actinobacillus pleuropneumoniae using cDNA microarrays

    OpenAIRE

    2007-01-01

    Abstract Background The bacterium Actinobacillus pleuropneumoniae is responsible for porcine pleuropneumonia, a widespread, highly contagious and often fatal respiratory disease of pigs. The general porcine innate immune response after A. pleuropneumoniae infection is still not clarified. The objective of this study was hence to characterise the transcriptional response, measured by using cDNA microarrays, in pigs 24 hours after experimental inoculation with A. pleuropneumoniae. Methods Micro...

  5. Antimicrobial effect of chlorine dioxide on Actinobacillus actinomycetemcomitans in diabetes mellitus rats treated with insulin

    Directory of Open Access Journals (Sweden)

    Tantin Ermawati

    2012-03-01

    Full Text Available Background: Periodontitis is a chronic inflammatory disease of periodontal tissues. Etiology of periodontal disease includes Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans which is the most predominant disease-causing bacteria found in the gingival sulcus. Periodontitis can be exacerbated by the systemic disease, such as diabetes mellitus considered as a metabolic disease characterized by hyperglycemia due to insulin deficiency. Treatment of periodontitis is then required in patients with type I diabetes to avoid radical reaction that can not only cause bleeding, but can also prevent infection, as a result, topical antimicrobial therapy and blood glucose control are required. Topical antimicrobial chlorine dioxide is a disinfectant that is effective in killing A. actinomycetemcomitans. Purpose: This study is aimed to determine the effects of topical antimicrobial chlorine dioxide gel or rinse on the number of A. actinomycetemcomitans in DM rats treated with insulin. Methods: 20 three month old male Wistar rats with weight of 170–200 grams were divided into four groups. First, periodontitis and DM were manipulated into all groups through aloksan injection with dose of 170 mg/kg. Those rats in group I were treated with insulin and chlorine dioxide gel, those in group II were treated with insulin and chlorine dioxide rinse, those in group III were treated with insulin only, and those in group IV were without treatment. In the third and seventh weeks, the number of A. actinomycetemcomitans was measured. The data was tested by using One-Way ANOVA test followed by LSD test. Results: The study showed that chlorine dioxide gel has a greater ability in reducing the number of A. actinomycetemcomitans than chlorine dioxide rinse although both are antimicrobials. Conclusion: It can be concluded that the use of chlorine dioxide gel can more effective to decrease the number of A. actinomycetemcomitans than chlorine dioxide rinse in DM rats

  6. Anticorpos antileucotoxina contra Actinobacillus actinomycetemcomitans em amostras de soro e saliva de pacientes com periodontite juvenil localizada Anti-leukotoxin antibodies against Actinobacillus actinomycetemcomitans in serum and saliva samples from patients with localized juvenile periodontitis

    Directory of Open Access Journals (Sweden)

    Roberto Issamu NAKAGAWA

    2001-03-01

    Full Text Available A leucotoxina de Actinobacillus actinomycetemcomitans é considerada seu principal fator de virulência com potencial de causar agressão às defesas do hospedeiro. No presente trabalho, foram analisados os níveis séricos e salivares de anticorpos antileucotoxina de A. actinomycetemcomitans em soros e salivas de pacientes com periodontite juvenil localizada (PJL e controles saudáveis. Adicionalmente, foi realizada a análise de complexo imune (CI nas amostras de saliva. Foram utilizados os métodos ELISA clássico com a leucotoxina obtida por gel filtração em Sephadex G-200 e ELISA de captura utilizando IgG de coelho anti-A. actinomycetemcomitans FDC Y4 leucotóxico adsorvido com uma cepa da mesma espécie, porém, não leucotóxica. Os resultados obtidos demonstraram níveis séricos de IgG significativamente mais elevados em pacientes com PJL em relação aos controles sadios, tanto por ELISA clássico como por ELISA de captura (p The leukotoxin produced by Actinobacillus actinomycetemcomitans is considered the major virulence factor with potential to cause damage to the host defenses. The present work analyzed the serumal and salivary levels of antibodies against the leukotoxin produced by A. Actinomycetemcomitans, in patients with Localized Juvenile Periodontitis (LJP and in healthy controls. Additionally, analysis of the immune complex (IC was carried out in saliva samples . The classic ELISA method, with leukotoxin obtained through Sephadex G-200 gel filtration, and the capture ELISA method, using rabbit anti-A. Actinomycetemcomitans (leucotoxic, FDC Y4, IgG adsorbed with a non-leukotoxic strain of A. actinomycetemcomitans, were used. The results obtained demonstrated significantly higher serumal levels of IgG in patients with LJP, when they were compared with the healthy controls, both for the classic and capture ELISA methods (p < 0.05. However, no significant differences were observed between the salivary levels of IgG, SIgA and IC

  7. 氧化还原电位对Actinobacillus succinogenes厌氧发酵生产丁二酸的影响%Effects of culture redox potential on succinic acid production by Actinobacillus succinogenes in anaerobic fermentation

    Institute of Scientific and Technical Information of China (English)

    周威; 郑璞; 倪晔; 姜岷; 韦萍; 孙志浩

    2008-01-01

    为提高琥珀酸放线菌Actinobacillus succinogenes CGMCC 1593厌氧发酵产丁二酸的水平,研究了以葡萄糖为C源,发酵液中不同氧化还原电位(VORP)对A.succinogenes CGMCC 1593生长和代谢产物分布的影响.结果表明:菌体生长和丁二酸积累的较佳VORP分别为-220 mV和-270 mV;利用代谢流分析法,比较VORP在-220 mV和-270 mV时发酵对数生长期(8 h)和稳定期(20 h)的代谢通量分布,以及发酵过程中磷酸烯醇式丙酮酸(PEP)、丙酮酸(Pyr)节点,NADH通量分配的变化,由此得出在VORP为-270 mV时,NADH总通量和丁二酸方向代谢通量增幅明显.在发酵过程中,通过降低VPRP至-270 mV,使丁二酸的产率从70%提高到85%.

  8. Use of corn steep liquor as an economical nitrogen source for biosuccinic acid production by Actinobacillus succinogenes

    Science.gov (United States)

    Tan, J. P.; Jahim, J. M.; Wu, T. Y.; Harun, S.; Mumtaz, T.

    2016-06-01

    Expensive raw materials are the driving force that leads to the shifting of the petroleum-based succinic acid production into bio-based succinic acid production by microorganisms. Cost of fermentation medium is among the main factors contributing to the total production cost of bio-succinic acid. After carbon source, nitrogen source is the second largest component of the fermentation medium, the cost of which has been overlooked for the past years. The current study aimed at replacing yeast extract- a costly nitrogen source with corn steep liquor for economical production of bio-succinic acid by Actinobacillus succinogenes 130Z. In this study, a final succinic acid concentration of 20.6 g/L was obtained from the use of corn steep liquor as the nitrogen source, which was comparable with the use of yeast extract as the nitrogen source that had a final succinate concentration of 21.4 g/l. In terms of economical wise, corn steep liquor was priced at 200 /ton, which was one fifth of the cost of yeast extract at 1000 /ton. Therefore, corn steep liquor can be considered as a potential nitrogen source in biochemical industries instead of the costly yeast extract.

  9. Utilization of CO2 fixating bacterium Actinobacillus succinogenes 130Z for simultaneous biogas upgrading and biosuccinic acid production.

    Science.gov (United States)

    Gunnarsson, Ingólfur B; Alvarado-Morales, Merlin; Angelidaki, Irini

    2014-10-21

    Biogas is an attractive renewable energy carrier. However, it contains CO2 which limits its use for certain applications. Here we report a novel approach for removing CO2 from biogas and capturing it as a biochemical through a biological process. This approach entails converting CO2 into biosuccinic acid using the bacterial strain Actinobacillus succinogenes 130 Z, and simultaneously producing high-purity CH4 (> 95%). Results showed that when pressure during fermentation was increased from 101.325 to 140 kPa, higher CO2 solubility was achieved, thereby positively affecting final succinic acid yield and titer, CO2 consumption rate, and CH4 purity. When using biogas as the only CO2 source at 140 kPa, the CO2 consumption rate corresponded to 2.59 L CO2 L(-1) d(-1) with a final succinic acid titer of 14.4 g L(-1). Under this pressure condition, the highest succinic acid yield and biogas quality reached corresponded to 0.635 g g(-1) and 95.4% (v v(-1)) CH4 content, respectively, after 24 h fermentation. This work represents the first successful attempt to develop a system capable of upgrading biogas to vehicle fuel/gas grid quality and simultaneously produce biosuccinic acid, a valuable building block with large market potential in the near term.

  10. Bagasse hydrolyzates from Agave tequilana as substrates for succinic acid production by Actinobacillus succinogenes in batch and repeated batch reactor.

    Science.gov (United States)

    Corona-González, Rosa Isela; Varela-Almanza, Karla María; Arriola-Guevara, Enrique; Martínez-Gómez, Álvaro de Jesús; Pelayo-Ortiz, Carlos; Toriz, Guillermo

    2016-04-01

    The aim of this work was to obtain fermentable sugars by enzymatic or acid hydrolyses of Agave tequilana Weber bagasse in order to produce succinic acid with Actinobacillus succinogenes. Hydrolyses were carried out with mineral acids (sulfuric and hydrochloric acids) or a commercial cellulolytic enzyme, and were optimized statistically by a response surface methodology, having as factors the concentration of acid/enzyme and time of hydrolysis. The concentration of sugars obtained at optimal conditions for each hydrolysis were 21.7, 22.4y 19.8g/L for H2SO4, HCl and the enzymatic preparation respectively. Concerning succinic acid production, the enzymatic hydrolyzates resulted in the highest yield (0.446g/g) and productivity (0.57g/Lh) using A. succinogenes in a batch reactor system. Repeated batch fermentation with immobilized A. succinogenes in agar and with the enzymatic hydrolyzates resulted in a maximum concentration of succinic acid of 33.6g/L from 87.2g/L monosaccharides after 5 cycles in 40h, obtaining a productivity of 1.32g/Lh.

  11. Evaluation of a single dose versus a divided dose regimen of amoxycillin in treatment of Actinobacillus pleuropneumoniae infection in pigs.

    Science.gov (United States)

    Lauritzen, B; Lykkesfeldt, J; Friis, C

    2005-08-01

    The theory of a time-dependent effect of amoxycillin was examined in a model of porcine Actinobacillus pleuropneumoniae (Ap)-infection using clinically relevant dosage regimens. Twenty hours after infection of fourteen pigs, when clinical signs of pneumonia were present, one group of pigs received a single dose of amoxycillin (20 mg/kg, i.m.), whereas another group received four doses of 5 mg/kg injected at 8-h intervals. A similar AUC of the plasma amoxycillin concentration versus time curve was obtained in the two groups, whereas the maximum concentration was threefold higher using the single high dose. Plasma amoxycillin was above the MIC for twice as long using the fractionated dosage scheme. The condition of the animals was evaluated by clinical and haematological observations combined with quantification of biochemical infection markers: C-reactive protein, zinc and ascorbic acid. Within 48 h of treatment, the pigs in both treatment groups recovered clinically. No significant differences in the time-course of clinical observations or plasma concentrations of the biomarkers of infection were observed between the two treatments. In conclusion, the efficacy of these two dosage regimens of amoxycillin was not significantly different in treatment of acute Ap-infection in pigs.

  12. Characterization of the omlA gene from different serotypes of Actinobacillus pleuropneumoniae: a new insight into an old approach

    Directory of Open Access Journals (Sweden)

    Ciro César Rossi

    2013-01-01

    Full Text Available The OmlA protein is a virulence factor of Actinobacillus pleuropneumoniae, an important pathogen in pigs. The polymorphisms present in the omlA gene sequence of 15 reference serotypes of A. pleuropneumoniae and non-serotypable isolates were assessed to determine the possible evolutionary relationship among them and to validate the importance of this gene as a molecular marker for the characterization of this bacterium. Divergence among the 15 serotypes of A. pleuropneumoniae probably resulted initially from two major evolutionary events that led to subsequent differentiation into nine groups. This differentiation makes it possible to characterize most of the serotypes by using bionformatics, thereby avoiding problems with immunological cross-reactivity. A conserved α-helix common to all the serotypes was most likely involved in connecting the protein to the outer membrane and acting as a signal peptide. A previously unknown gene duplication was also identified and could contribute to the genetic variability that makes it difficult to serotype some isolates. Our data support the importance of the omlA gene in the biology of A. pleuropneumoniae and provide a new area of research into the OmlA protein.

  13. ICEApl1, an integrative conjugative element related to ICEHin1056, identified in the pig pathogen Actinobacillus pleuropneumoniae

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    Janine T Bosse

    2016-06-01

    Full Text Available ICEApl1 was identified in the whole genome sequence of MIDG2331, a tetracycline-resistant (MIC = 8 mg/L serovar 8 clinical isolate of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. PCR amplification of virB4, one of the core genes involved in conjugation, was used to identify other A. pleuropneumoniae isolates potentially carrying ICEApl1. MICs for tetracycline were determined for virB4 positive isolates, and shotgun whole genome sequence analysis was used to confirm presence of the complete ICEApl1. The sequence of ICEApl1 is 56083 bp long and contains 67 genes including a Tn10 element encoding tetracycline resistance. Comparative sequence analysis was performed with similar integrative conjugative elements (ICEs found in other members of the Pasteurellaceae. ICEApl1 is most similar to the 59393 bp ICEHin1056, from Haemophilus influenzae strain 1056. Although initially identified only in serovar 8 isolates of A. pleuropneumoniae (31 from the UK and 1 from Cyprus, conjugal transfer of ICEApl1 to representative isolates of other serovars was confirmed. All isolates carrying ICEApl1 had a MIC for tetracycline of 8 mg/L. This is, to our knowledge, the first description of an ICE in A. pleuropneumoniae, and the first report of a member of the ICEHin1056 subfamily in a non-human pathogen. ICEApl1 confers resistance to tetracycline, currently one of the more commonly used antibiotics for treatment and control of porcine pleuropneumonia.

  14. ICEApl1, an Integrative Conjugative Element Related to ICEHin1056, Identified in the Pig Pathogen Actinobacillus pleuropneumoniae

    Science.gov (United States)

    Bossé, Janine T.; Li, Yanwen; Fernandez Crespo, Roberto; Chaudhuri, Roy R.; Rogers, Jon; Holden, Matthew T. G.; Maskell, Duncan J.; Tucker, Alexander W.; Wren, Brendan W.; Rycroft, Andrew N.; Langford, Paul R.

    2016-01-01

    ICEApl1 was identified in the whole genome sequence of MIDG2331, a tetracycline-resistant (MIC = 8 mg/L) serovar 8 clinical isolate of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. PCR amplification of virB4, one of the core genes involved in conjugation, was used to identify other A. pleuropneumoniae isolates potentially carrying ICEApl1. MICs for tetracycline were determined for virB4 positive isolates, and shotgun whole genome sequence analysis was used to confirm presence of the complete ICEApl1. The sequence of ICEApl1 is 56083 bp long and contains 67 genes including a Tn10 element encoding tetracycline resistance. Comparative sequence analysis was performed with similar integrative conjugative elements (ICEs) found in other members of the Pasteurellaceae. ICEApl1 is most similar to the 59393 bp ICEHin1056, from Haemophilus influenzae strain 1056. Although initially identified only in serovar 8 isolates of A. pleuropneumoniae (31 from the UK and 1 from Cyprus), conjugal transfer of ICEApl1 to representative isolates of other serovars was confirmed. All isolates carrying ICEApl1 had a MIC for tetracycline of 8 mg/L. This is, to our knowledge, the first description of an ICE in A. pleuropneumoniae, and the first report of a member of the ICEHin1056 subfamily in a non-human pathogen. ICEApl1 confers resistance to tetracycline, currently one of the more commonly used antibiotics for treatment and control of porcine pleuropneumonia. PMID:27379024

  15. Utilization of CO2 fixating bacterium Actinobacillus succinogenes 130Z for simultaneous biogas upgrading and bio-succinic acid production

    DEFF Research Database (Denmark)

    Gunnarsson, Ingólfur Bragi; Alvarado-Morales, Merlin; Angelidaki, Irini

    2014-01-01

    acid using the bacterial strain Actinobacillus succinogenes 130Z, and simultaneously producing high purity CH4 (>95%). Results showed that when pressure during fermentation was increased from 101.325 to 140 kPa, higher CO2 solubility was achieved, thereby positively affecting final succinic acid yield...... and titre, CO2 consumption rate and CH4 purity. When using biogas as the only CO2 source at 140 kPa, the CO2 consumption rate corresponded to 2.59 L CO2 L-1 d-1 with a final succinic acid titre of 14.4 g L-1. Under this pressure condition the highest succinic acid yield and biogas quality reached...... corresponded to 0.635 g g-1 and 95.4% (v v-1) CH4 content respectively after 24 hours fermentation. This work represents the first successful attempt to develop a system capable of upgrading biogas to vehicle fuel/gas grid quality and simultaneously produce bio-succinic acid, a valuable building block...

  16. Evaluation of an enzyme-linked immunosorbent assay for serological surveillance of infection with Actinobacillus pleuropneumoniae serotype 5 in pig herds

    DEFF Research Database (Denmark)

    Klausen, Joan; Andresen, Lars Ole; Barfod, Kristen;

    2002-01-01

    An indirect enzyme-linked immunoassay for serological surveillance of infection of pigs with Actinobacillus pleuropneumoniae (Ap) serotype 5 was developed. The antigen used was prepared from Ap serotype 5b strain L20. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis...... showed that the antigen contained high molecular weight lipopolysaccharide (LPS) and presumably also capsular polysaccharide (CP). The Ap serotype 5 ELISA was tested using sera from pigs experimentally infected with the 12 different Ap serotypes of biotype 1 and with sera from herds naturally infected...

  17. Otimização da técnica da PCR para a detecção de Actinobacillus pleuropneumoniae Optimization of PCR technique for detection of Actinobacillus pleuropneumoniae

    Directory of Open Access Journals (Sweden)

    Karina Koerich de Souza

    2008-11-01

    Full Text Available A utilização de métodos moleculares baseados em PCR é fundamental na detecção do Actinobacillus pleuropneumoniae, sendo capaz de identificar a infecção antes do estabelecimento da doença no rebanho. Estes métodos apresentam maior sensibilidade quando comparados com métodos tradicionais de isolamento bacteriano, mas podem sofrer influência de substâncias que reduzem a especificidade do teste e proporcionam o aparecimento de amplificações inespecíficas. No intuito de reduzir as amplificações inespecíficas, observadas quando aplicada a PCR para o gene cpx em amostras de tecido tonsilar, procedeu-se a otimização da técnica, na qual foram analisados o efeito do pré-cultivo bacteriano e as diferentes temperaturas de anelamento dos iniciadores e foi introduzido, no protocolo, um anticorpo que se liga na enzima Taq DNA Polimerase, aumentando a especificidade do teste. Paralelamente, foi realizado um experimento para verificar o efeito inibidor do tecido tonsilar sobre os resultados da PCR. Para isso, porções de tonsila de animais negativos para A. pleuropneumoniae foram contaminadas artificialmente com a amostra referência do sorotipo 5B. A adição do anticorpo para a enzima Taq DNA Polimerase e o aumento da temperatura de anelamento dos iniciadores para 57°C diminuiu o aparecimento de amplificações inespecíficas. Os resultados obtidos no experimento demonstraram que o tecido tonsilar possui efeito inibidor nas amplificações da PCR. Além disso, a amplificação depende de, no mínimo, 675 UFC presentes na alíquota da amostra usada na PCR (equivalente a 1,35 x 10(5 UFC mL-1, assim, amostras de fragmentos de tecido de infecções iniciais e/ou com poucas células podem apresentar resultados falsos negativos.The use of molecular methods based on PCR is important in Actinobacillus pleuropneumoniae detection, being able to identify the infection before the establishment of the disease in the herd. These methods have larger

  18. LEUKOTOXIC ACTIVITY OF ACTINOBACILLUS ACTINOMYCETEMCOMITANS ISOLATED FROM HUMAN AND NON-HUMAN PRIMATES Atividade leucotóxica de amostras de Actinobacillus actinomycetemcomitans de primatas humanos não-humanos

    Directory of Open Access Journals (Sweden)

    Francisca Lúcia de Lima

    2001-10-01

    Full Text Available Actinobacillus actinomycetemcomitans is a clinically relevant periodontopathogenic Gram-negative coccobacillus that produces a leukotoxin of the RTX cytolysin family. In this study, we evaluated the leukotoxic activity of A. actinomycetemcomitans strains isolated from human and marmosets by Trypan blue exclusion and by the chemiluminescence assays. Among eight A. actinomycetemcomitans human strains studied, two (P2.17 and P8.12 were classified as high leukotoxin producers and among eight marmoset strains, one (M22.11 showed high leukotoxin production, as determined by Trypan blue exclusion assay. The reference strains ATCC 29523 and FDC Y4 respectively behaved like moderate and low producers. The chemiluminescence assay was used to evaluate the leukotoxic activity of M22.11 and P2.17 strains submitted to different growth conditions. Leukotoxic activity was detected on cells at the logarithmic phase and was similar under anaerobic and microaerophilic growth conditions. It was greatly reduced when cells were grown at glucose concentrations lower or higher than 0.75% (0.25% and 1.5% in thioglycolate medium. Leukotoxin production mainly by the M22.11 strain was low in BHI broth, whereas production in TSB medium showed a similar level as in thioglycolate broth medium. Sodium bicarbonate at 10 mM did not affect leukotoxin production.Actinobacillus actinomycetemcomitans é um cocobacilo Gram negativo, periodontopatógeno clinicamente importante, que produz uma leucotoxina pertencente à família das citolisinas RTX. Neste estudo, avaliou-se a atividade leucotóxica de amostras de A. actinomycetemcomitans isoladas de seres humanos e de calitriquídeos pelos métodos de exclusão de azul de Tripan e quimioluminescência. Duas (P2.17 e P8.12 entre oito amostras de A. actinomycetemcomitans isoladas de seres humanos, e uma (M22.11 entre 8 amostras isoladas de sagüis se apresentaram como altamente produtoras de leucotoxina, como determinado pelo teste de

  19. Diversidad genética de cepas de Actinobacillus pleuropneumoniae (App aisladas desde planteles de producción intensiva de cerdos en Chile Genetic diversity of Actinobacillus pleuropneumoniae (App strains in intensive swine farms in Chile

    Directory of Open Access Journals (Sweden)

    V Neira-Ramírez

    2012-01-01

    Full Text Available Actinobacillus pleuropneumoniae (App es el agente etiológico de la pleuroneumonía contagiosa porcina, una de las enfermedades de etiología bacteriana de mayor relevancia en producción porcina. En el mundo se han descrito 15 serotipos de App, en Chile solo los serotipos 1 y 5. La serotipificación requiere mucho tiempo, trabajo y dinero, actualmente se encuentran herramientas moleculares para realizar una "serotipificación" mediante la genotipificación de toxinas Apx. Así, se evaluaron 60 aislados de App provenientes de nueve empresas porcinas de producción intensiva distribuidas en distintas regiones de Chile, obtenidas desde pulmones de cerdos con lesiones compatibles con pleuroneumonía contagiosa porcina. Las bacterias fueron aisladas mediante los métodos tradicionales y confirmados por API, recolectados durante los años 2007, 2008 y 2009. Los resultados identificaron los genotipos correspondientes sólo a los serotipos 4, 6 y 7, los cuales se describen por primera vez en Chile, siendo el más frecuente el serotipo 7. En las diferentes zonas estudiadas, no existió un serotipo predominante, excepto en las regiones de O'Higgins y del Biobío en las cuales fue más frecuentemente aislado el serotipo 7. El presente estudio es el primer acercamiento con el fin de conocer la distribución de serotipos de App en Chile. Con el fin de conocer la real diversidad genética y serotipos de App en los diversos planteles en Chile es necesario realizar estudios que contemplen un mayor número de aislados.Actinobacillus pleuropneumoniae (App is the etiologic agent of porcine contagious pleuropneumonia, an important bacterial disease in intensive pig production. In the world were described 15 App serotypes, in Chile serotypes 1 and 5 have been reported. The serotyping technique is slow, expensive and difficult; currently, a molecular tool named PCR is available to "serotyping" by Apx toxins genotyping, which is quick, non-expensive and easy. 60 App

  20. Transcriptional portrait of Actinobacillus pleuropneumoniae during acute disease--potential strategies for survival and persistence in the host.

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    Kirstine Klitgaard

    Full Text Available BACKGROUND: Gene expression profiles of bacteria in their natural hosts can provide novel insight into the host-pathogen interactions and molecular determinants of bacterial infections. In the present study, the transcriptional profile of the porcine lung pathogen Actinobacillus pleuropneumoniae was monitored during the acute phase of infection in its natural host. METHODOLOGY/PRINCIPAL FINDINGS: Bacterial expression profiles of A. pleuropneumoniae isolated from lung lesions of 25 infected pigs were compared in samples taken 6, 12, 24 and 48 hours post experimental challenge. Within 6 hours, focal, fibrino hemorrhagic lesions could be observed in the pig lungs, indicating that A. pleuropneumoniae had managed to establish itself successfully in the host. We identified 237 differentially regulated genes likely to encode functions required by the bacteria for colonization and survival in the host. This group was dominated by genes involved in various aspects of energy metabolism, especially anaerobic respiration and carbohydrate metabolism. Remodeling of the bacterial envelope and modifications of posttranslational processing of proteins also appeared to be of importance during early infection. The results suggested that A. pleuropneumoniae is using various strategies to increase its fitness, such as applying Na+ pumps as an alternative way of gaining energy. Furthermore, the transcriptional data provided potential clues as to how A. pleuropneumoniae is able to circumvent host immune factors and survive within the hostile environment of host macrophages. This persistence within macrophages may be related to urease activity, mobilization of various stress responses and active evasion of the host defenses by cell surface sialylation. CONCLUSIONS/SIGNIFICANCE: The data presented here highlight the importance of metabolic adjustments to host conditions as virulence factors of infecting microorganisms and help to provide insight into the mechanisms

  1. The Adh adhesin domain is required for trimeric autotransporter Apa1-mediated Actinobacillus pleuropneumoniae adhesion, autoaggregation, biofilm formation and pathogenicity.

    Science.gov (United States)

    Wang, Lei; Qin, Wanhai; Yang, Shuxin; Zhai, Ruidong; Zhou, Liang; Sun, Changjiang; Pan, Fengguang; Ji, Qun; Wang, Yu; Gu, Jingmin; Feng, Xin; Du, Chongtao; Han, Wenyu; Langford, P R; Lei, Liancheng

    2015-05-15

    Actinobacillus pleuropneumoniae is a causative agent of porcine pleuropneumonia, which is a highly contagious endemic disease of pigs. Adhesion is a critical first step in the infection process. Trimeric autotransporter adhesions (TAAs) have been identified as novel virulence factors; however, little is known on their roles in A. pleuropneumoniae pathogenicity. Here, our data show that YadA-like head region (Adh) of Apa1 was the optimal adhesion functional domain via segment expression and adhesion assays in vitro. Additionally, Adh induced partial protection against A. pleuropneumoniae 5b L20 and serotypes 1, 3, and 5a in mice. The deletion of Adh gene significantly decreased autoaggregation, biofilm formation and adherence to host cells in vitro. Furthermore, with delaying of clinical symptoms, reducing production of pro-inflammatory cytokines and lessening the lung injury after infection, Adh deletion strain (5bϕAdh) significantly reduced the pathogenicity to piglets. To elucidate the mechanism of lung injury, the differentially expressed genes in the lung tissues of piglets infected with the 5b L20 or 5bϕAdh strains were investigated using microarray analysis and validated by qRT-PCR. Compared with the 5b L20 infected piglets, 495 genes were differentially expressed in 5bϕAdh infected lung tissue (221 upregulated and 274 downregulated). Especially, the antigen processing and presentation gene IFI30 was increased following infection with the 5bϕAdh strain. Thus, Adh may enhance pathogenicity by depressing host immune recognition. We conclude that the head domain of the A. pleuropneumoniae trimeric autotransporter Apa1 regulates autoagglutination, biofilm formation, adhesion to host cells and pathogenicity.

  2. Association between coinfection of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Treponema denticola and periodontal tissue destruction in chronic periodontitis

    Institute of Scientific and Technical Information of China (English)

    CHEN Li-li; WU Yan-min; YAN Jie; SUN Wei-lian; SUN Yu-zheng; David Ojcius

    2005-01-01

    Background The association between the infection of Porphyromonas gingivalis, Actinobacillus actinomy-cetemcomitans and Treponema denticola in chronic periodontitis (CP) and the severity of periodontal disease remains to be elucidated. The aim of this study was to investigate the subgingival infection frequencies of three periodontopathic bacteria in Chinese CP patients and to evaluate the correlations between infection by these bacteria and periodontal destruction.Methods A multiple PCR assay using primers derived from 16SrDNA genes of P. gingivalis, A. actinomy-cetemitans and T. denticola was established to measure simultaneously the presence of the three microbes in 162 subgingival samples from 81 Chinese CP patients. Results The positive rates of P. gingivalis, A. actinomycetemitans and T. denticola in the subgingival samples were 84.6%, 83.3% and 88.3%, respectively. Of the subgingival samples, 68% revealed the coinfection of all the three microbes. The infection rates with P. gingivalis, A. actinomycetemitans or T. denticola alone was 5.9% (1/17), 17.6% (3/17) and 76.5% (13/17), respectively. A close association was present between the A. actinomycetemitans infection and gingival index (GI) (P0.05). P. gingivalis and A. actinomycetemitans were more frequently detectable in middle and deep pockets than in shallow ones (P<0.01), while T. denticola was found remarkably often in deep pockets (P<0.05). The coinfection rate of the three microbes was significantly higher in sites with severe periodontitis than in those with mild periodontitis (P<0.01). Conclusions The multiple PCR established in this study can be used as a sensitive and specific method to simultaneously detect all three microbes in subgingival samples. A. actinomycetemitans infection may be associated with CP and play an important role in the periodontal tissue destruction. The coinfection of P. gingivalis, A. actinomycetemitans and T. denticola can cause more serious periodontal destruction than

  3. Expression levels of immune markers in Actinobacillus pleuropneumoniae infected pigs and their relation to breed and clinical symptoms

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    Rothkoetter Hermann-Josef

    2009-04-01

    Full Text Available Abstract Background In pigs little is known about the role of innate immune defence in bacterial infections of the respiratory tract, despite their major role in pig production. In the present study we characterized and compared in vitro and in vivo activation of immune markers of different pig breeds 7 days before, and 4 and 21 days after an experimental aerosol infection with Actinobacillus (A. pleuropneumoniae. Results In vitro stimulation of bronchoalveolar lavage fluid (BALF and blood leukocytes with A. pleuropneumoniae, Streptococcus suis, PMA and LPS led to production of different amounts of H2O2, NO and TNF-α, depending on the stimulus, individual, breed and time of infection. Generally, significant responses to in vitro stimulation were observed only in blood leukocytes, whereas the alveolar macrophages showed a high basal activation. In addition, the production of haptoglobin and cytokines (TNF-α, IFN-γ and IL-10 in vivo was measured in plasma and BALF. Plasma haptoglobin levels mirrored the clinical manifestations at 4 days post-infection. In plasma and BALF TNF-α could not be detected, whereas variable levels of IFN-γ were found at pre- and post-infection times. IL-10 was found in some plasma but in none of the BALF samples. The different expression levels in individuals within the breeds correlated for some markers with the severity of clinical manifestations, e.g. H2O2, plasma haptoglobin and BALF IFN-γ for German Landrace pigs. Conclusion Our findings revealed differences in the activation of the immune markers with respect to infection time, individuals and breeds. Moreover, results showed different correlation grades between the immune markers produced in vitro or in vivo and the clinical manifestations. Further analyses will have to show whether these markers may serve as correlates of protection against porcine respiratory infections.

  4. Porcine mononuclear phagocyte subpopulations in the lung, blood and bone marrow: dynamics during inflammation induced by Actinobacillus pleuropneumoniae

    Science.gov (United States)

    Ondrackova, Petra; Nechvatalova, Katerina; Kucerova, Zdenka; Leva, Lenka; Dominguez, Javier; Faldyna, Martin

    2010-01-01

    Mononuclear phagocytes (MP) are cells of nonspecific immunity, playing an essential role in defense against bacterial pathogens. Although various MP subpopulations have been described in the pig, relations among these populations in vivo are unknown to date. The present study was aimed at describing porcine MP subpopulations infiltrating inflamed tissue of pigs under in vivo conditions. Actinobacillus pleuropneumoniae (APP) infection was used to induce an inflammatory response. CD172α, CD14, CD163, MHCII and CD203α cell surface molecules were used to identify MP by flow cytometry. Changes in MP subpopulations in the peripheral blood (PB) and bone marrow (BM) compartments along with the analysis of MP appearing in the inflamed lungs were assessed to elucidate the possible origin and maturation stages of the infiltrating MP. The MP population migrating to the inflamed lungs was phenotype CD14+ CD163+ CD203α+/− MHCII+/−. Concomitantly, after APP infection there was an increase in the PB MP CD14+ CD163+ CD203α− MHC II− population, suggesting that these cells give rise to inflammatory monocytes/macrophages. The CD203α and MHCII molecules appear on these cells after leaving the PB. In healthy animals, the BM MP precursors were represented by CD14− CD163− cells maturing directly into CD14+ CD163− that were then released into the PB. After infection, an altered maturation pathway of MP precursors appeared, represented by CD14− CD163− CD203α− MHCII− MP directly switching into CD14+ CD163+ CD203α− MHCII− MP. In conclusion, two different MP maturation pathways were suggested in pigs. The use of these pathways differs under inflammatory and noninflammatory conditions. PMID:20519113

  5. Probing of Actinobacillus pleuropneumoniae ApxIIIA toxin-dependent cytotoxicity towards mammalian peripheral blood mononucleated cells

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    Fett Thomas

    2008-12-01

    Full Text Available Abstract Background Actinobacillus pleuropneumoniae, the causative bacterial agent of porcine pleuropneumonia, produces Apx toxins which belong to RTX toxin family and are recognized as the major virulence factors. So far, their target receptor(s has not been identified and the disease cytopathogenesis remains poorly understood. Production of an active Apx toxin and characterization of its toxic activity constitute the premises necessary to the description of its interaction with a potential receptor. From this point of view, we produced an active recombinant ApxIIIA toxin in order to characterize its toxicity on peripheral blood mononucleated cells (PBMCs isolated from several species. Findings Toxin preparation exercises a strong cytotoxic action on porcine PBMCs which is directly related to recombinant ApxIIIA since preincubation with polymyxin B does not modify the cytotoxicity rate while preincubation with a monospecific polyclonal antiserum directed against ApxIIIA does. The cell death process triggered by ApxIIIA is extremely fast, the maximum rate of toxicity being already reached after 20 minutes of incubation. Moreover, ApxIIIA cytotoxicity is species-specific because llama, human, dog, rat and mouse PBMCs are resistant. Interestingly, bovine and caprine PBMCs are slightly sensitive to ApxIIIA toxin too. Finally, ApxIIIA cytotoxicity is cell type-specific as porcine epithelial cells are resistant. Conclusion We have produced an active recombinant ApxIIIA toxin and characterized its specific cytotoxicity on porcine PBMCs which will allow us to get new insights on porcine pleuropneumonia pathogenesis in the future.

  6. A 24-kDa cloned zinc metalloprotease from Actinobacillus pleuropneumoniae is common to all serotypes and cleaves actin in vitro.

    Science.gov (United States)

    García-Cuéllar, C; Montañez, C; Tenorio, V; Reyes-Esparza, J; Durán, M J; Negrete, E; Guerrero, A; de la Garza, M

    2000-01-01

    Actinobacillus pleuropneumoniae causes pleuropneumonia in swine. This bacterium secretes proteases that degrade porcine hemoglobin and IgA in vitro. To further characterize A. pleuropneumoniae proteases, we constructed a genomic library expressed in Escherichia coli DH5alpha, and selected a clone that showed proteolytic activity. The recombinant plasmid carries an 800-base pair A. pleuropneumoniae gene sequence that.codes for a 24-kDa polypeptide. A 350-base pair PstI fragment from the sequence hybridized at high stringency with DNA from 12 serotypes of A. pleuropneumoniae, but not with DNA from Actinobacillus suis, Haemophilus parasuis, Pasteurella haemolytica, Pasteurella multocida A or D, or E. coli DH5alpha, thus showing specificity for A. pleuropneumoniae. The expressed polypeptide was recognized as an antigen by convalescent-phase pig sera. Furthermore, a polyclonal antiserum developed against the purified polypeptide recognized an A. pleuropneumoniae oligomeric protein in both crude-extract and cell-free culture media. This recombinant polypeptide cleaved azocoll, gelatin, and actin. Inhibition of the proteolytic activity by diethylpyrocarbonate suggests that this polypeptide is a zinc metalloprotease. Images Figure 1. Figure 2. Figure 3. Figure 4. Figure 6. Figure 7. PMID:10805246

  7. Field experience with two different vaccination strategies aiming to control infections with Actinobacillus pleuropneumoniae in a fattening pig herd

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    Sjölund Marie

    2010-03-01

    Full Text Available Abstract Background The prevalence of pleurisies recorded at slaughter is increasing in Sweden, and acute outbreaks of actinobacillosis that require antimicrobial treatments have become more frequent. As an increased use of antimicrobials may result in the development of antimicrobial resistance it is essential to develop alternative measures to control the disease. Vaccinations present an appealing alternative to antimicrobial treatments. The aim of this work was to evaluate the potential of two different vaccination strategies in a specialized fattening herd affected by actinobacillosis. Methods The study was conducted in a specialized fattening herd employing age segregated rearing in eight units. The herd suffered from infections caused by Actinobacillus pleuropneumoniae serotype 2, confirmed by necropsy and serology. The study included 54 batches of pigs grouped into five periods. Batches of pigs of the second period were vaccinated against actinobacillosis twice, and pigs in the fourth period were vaccinated three times. Batches of pigs of the first, third and fifth period were not vaccinated. Concentrations of serum antibodies to A. pleuropneumoniae and serum amyloid A (SAA were analysed and production data were recorded. Results Despite vaccinating, medical treatments were required to reduce the impact of the disease. The mean incidence of individual treatments for respiratory diseases during the rearing period ranged from 0 to 4.7 ± 1.8%, and was greatest during the triple vaccination period (period IV; p A. pleuropneumoniae serotype 2 in the absence of a SAA-response. The prevalence of pleuritis decreased from 25.4 ± 6.5% in the first period to 5.0 ± 3.7% in the fifth period (p Conclusions The vaccine did not effectively prevent clinical expression of A. pleuropneumoniae infections, but seroconversion to A. pleuropneumoniae in the absence of a SAA-response in a large number pigs indicated that the vaccine had activated the immune

  8. Comparative profiling of the transcriptional response to iron restriction in six serotypes of Actinobacillus pleuropneumoniae with different virulence potential

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    Angen Øystein

    2010-12-01

    Full Text Available Abstract Background Comparative analysis of gene expression among serotypes within a species can provide valuable information on important differences between related genomes. For the pig lung pathogen Actinobacillus pleuropneumoniae, 15 serotypes with a considerable variation in virulence potential and immunogenicity have been identified. This serotypic diversity can only partly be explained by amount of capsule and differences in the RTX toxin genes in their genomes. Iron acquisition in vivo is an important bacterial function and in pathogenic bacteria, iron-limitation is often a signal for the induction of virulence genes. We used a pan-genomic microarray to study the transcriptional response to iron restriction in vitro in six serotypes of A. pleuropneumoniae (1, 2, 3, 5b, 6, and 7, representing at least two levels of virulence. Results In total, 45 genes were significantly (p A. pleuropneumoniae was the up-regulation of a putative cirA-like siderophore in all six serotypes. Three genes, recently described in A. pleuropneumoniae as possibly coding for haemoglobin-haptoglobin binding proteins, displayed significant serotype related up-regulation to iron limitation. For all three genes, the expression appeared at its lowest in serotype 3, which is generally considered one of the least virulent serotypes of A. pleuropneumoniae. The three genes share homology with the hmbR haemoglobin receptor of Neisseria meningitidis, a possible virulence factor which contributes to bacterial survival in rats. Conclusions By comparative analysis of gene expression among 6 different serotypes of A. pleuropneumoniae we identified a common set of presumably essential core genes, involved in iron regulation. The results support and expand previous observations concerning the identification of new potential iron acquisition systems in A. pleuropneumoniae, showing that this bacterium has evolved several strategies for scavenging the limited iron resources of the

  9. Analysis of leukotoxin gene types of Actinobacillus actinomycetemcomitans in brazilians with aggressive periodontitis Análise de genes da leucotoxina de Actinobacillus actinomycetemcomitans em brasileiros com periodontite agressiva

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    Wilson Rosalem Junior

    2006-06-01

    Full Text Available Actinobacillus actinomycetemcomitans (Aa is considered a major etiologic agent of aggressive periodontitis but this species has also been associated with other forms of periodontal disease. Further, highly leukotoxic strains are related to severity of disease. The purpose of this study was to determine the prevalence of Aa and the occurrence of the leukotoxin gene 530-bp deletion in patients with generalized aggressive periodontitis (GAP from a Brazilian population. Thirty periodontally healthy and 29 GAP subjects participated in the study. Full-mouth periodontal examination including probing pocket depth (PPD, clinical attachment level (CAL, supragingival biofilm (SB and bleeding on probing (BOP was carried out at 6 sites/tooth for all subjects. Whole saliva samples were collected for bacterial DNA isolation. The detection of Aa and the presence of the 530-bp genetic deletion were determined directly in the samples by polimerase chain reaction. Differences on clinical and microbiological parameters between the two groups were sought using the Mann-Whitney, Fisher´s exact and Chi-square tests. Associations between clinical and microbiological parameters were tested using the Pearson correlation coefficient. Aa was detected significantly more often in GAP patients (96.6% than healthy subjects (76.7% (chi2 = 4.9; p Actinobacillus actinomycetemcomitans (Aa tem sido associado com diferentes formas de doenças periodontais, mas tal espécie é considerada o principal agente etiológico da doença periodontal agressiva. Algumas cepas de Aa apresentam uma deleção de 530 pb na região promotora do operon do gene da leucotoxina, produzindo assim maiores quantidades de leucotoxina. Tal fato pode ter um importante papel na patogênese das doenças periodontais. A proposta do presente estudo foi determinar a prevalência do Aa e a ocorrência da deleção genética da leucotoxina em pacientes com periodontite agressiva generalizada (PAG de uma amostra da

  10. Development and evaluation of a mixed long-chain lipopolysaccharide based ELISA for serological surveillance of infection with Actinobacillus pleuropneumoniae serotypes 2, 6 and 12 in pig herds

    DEFF Research Database (Denmark)

    Grøndahl-Hansen, Jan; Barfod, Kristen; Klausen, Joan;

    2003-01-01

    The objective was to develop an enzyme-linked immunosorbent assay (ELISA) for simultaneous detection of antibodies against Actinobacillus pleuropneumoniae (Ap) serotypes 2, 6 and 12. The assay was designated MIX-ELISA. Lipopolysaccharide (LPS) from Ap serotypes 2, 6 and 12 was purified using hot...... phenol-water extraction followed by fractionation by size-exclusion chromatography. A mixture of fractions containing molecules with molecular weight above 50 kDa from all three serotypes was used as antigen. The MIX-ELISA was evaluated with sera from pigs experimentally infected with the serotypes 1, 2......, 5b, 6, 7, 8, 10 and 12 of Ap biotype 1. In addition to reaction with sera from pigs inoculated with Ap serotypes 2, 6 and 12, reaction was observed with sera from pigs inoculated with serotype 8. Furthermore, the sensitivity and specificity of the test on a herd level were evaluated with sera from...

  11. Validation of putative reference genes for qRT-PCR normalization in tissues and blood from pigs infected with Actinobacillus pleuropneumoniae

    DEFF Research Database (Denmark)

    Skovgaard, Kerstin; Mortensen, Shila; Poulsen, K.T.;

    2007-01-01

    The quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) is a sensitive and very efficient technique for quantification of gene expression. However, qRT-PCR relies on accurate normalization of gene expression data, as RNA recovery and cDNA synthesis efficiency might vary...... from sample to sample. In the present study, six putative reference genes were validated for normalization of gene expression in three different tissues and in white blood cells from pigs experimentally infected with the common respiratory pathogen Actinobacillus pleuropneumoniae. Two dedicated...... (GAPDH). IL-6 expression was quantified in white blood cells, liver, lymph nodes and tonsils from 10 infected pigs and 5 control pigs. After normalization using either geNorm or Normfinder IL-6 was shown to be significantly up-regulated (P

  12. Incidence of Reinfections with Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae in Pig Farms Located in Respiratory-Disease-Free Regions of Switzerland – Identification and Quantification of Risk Factors

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    Scheidegger R

    2002-09-01

    Full Text Available The objective of the study was to identify risk factors for reintroduction of Actinobacillus pleuopneumoniae and Mycoplasma hyopneumoniae (enzootic pneumonia onto pig farms in areas in Switzerland that were involved in an eradication programme from 1996 to 1999 and to assess the role of dealers in relation to these reinfections. The study was based on the comparison of pig farms that were reinfected in the year 2000 (cases and pig farms that remained uninfected in the same area (controls. Additionally, data were collected from Swiss pig dealers and transport companies. Out of a total of 3983 farms, 107 farms were reinfected in the year 2000. The incidences were 0.1% for Actinobacillus pleuopneumoniae and 2.6% for Mycoplasma hyopneumoniae (enzootic pneumonia. Compared to reinfection rates prior to the eradication programme, this is a considerable reduction. Statistically significant risk factors for the reinfection were 'finishing farm', 'large mixed breeding-finishing farm', 'reinfected neighbour' and 'parking site for pig transport vehicles close to the farm'. Pig farmers that purchased pigs from only one supplier per batch had a lower risk of reintroducing infection (protective factor. As long as infected and uninfected regions co-exist in Switzerland, direct and indirect contact between farms, pig herds and slaughter sites via transport vehicles are a major pathway of disease spread. Risk management measures linked to these contacts are therefore of key importance. The survey of dealers indicated various areas for improvement such as strategic planning of pick-up routes or cleaning and disinfecting of trucks.

  13. Estudio del comportamiento serológico de Actinobacillus pleuropneumoniae (App en planteles porcinos comerciales de la zona central de Chile Serological behaviour study of Actinobacillus pleuropneumoniae (App in commercial swine herds from the central region of Chile

    Directory of Open Access Journals (Sweden)

    D Muñoz

    2008-01-01

    Full Text Available En Chile se ha realizado sólo un estudio en Actinobacillus pleuropneumoniae (App. Este trabajo pretende determinar la duración de la inmunidad materna, la edad de seroconversión y la prevalencia aparente y verdadera en 7 planteles de cerdos comerciales. Se obtuvieron 60 muestras por plantel, divididas en 10 muestras de suero, de animales de 4, 6, 10, 14,18 y 21 semanas de edad, y analizadas a través de un kit ELISA® comercial. De las 420 muestras se detectaron 134 positivas, de las cuales 112 correspondían a cerdos menores de 10 semanas y sólo 22 provenían de animales mayores de 10 semanas, que seroconvirtieron probablemente debido a una infección de campo. La caída de la inmunidad materna fue alrededor de la 10ª semana de edad. En cuanto a la seroconversión, se observó que a partir de la 18* semana comenzaron a aparecer los animales con anticuerpos circulantes propios. Dos de los siete planteles no seroconvirtieron. Además, dos presentaron una seroconversión igual o superior al 50% a las 18 semanas. La seroprevalencia aparente de App fue de 10,48%, mientras que prevalencia verdadera, mediante dos métodos estadísticos, fue de 9,6% (IC: 7,6% y 11,7% y 10,67% respectivamente. En este trabajo se encontró que la prevalencia es similar a la observada en EE.UU., debido presumiblemente al sistema de producción y a los serotipos que están presentes en ambos países. Por otro lado, si bien la mayoría de los planteles seroconvierten luego de la caída de la inmunidad materna, se observaron diferentes patrones serológicos entre ellos.In Chile, there was only one existing study on App. This study was designed to determine the maternal immunity duration, the age of seroconversion and the apparent and true prevalence in animals from 7 swine commercial herds. 60 samples were taken per herd and divided into 10 serum samples from animals of 4, 6,10,14,18and21 weeks of age, which were analyzed by ELISA®. Out of the 420 samples, 134 were

  14. Real-time quantitative reverse transcription-PCR analysis of expression stability of Actinobacillus pleuropneumoniae housekeeping genes during in vitro growth under iron-depleted conditions

    DEFF Research Database (Denmark)

    Nielsen, K. K.; Boye, Mette

    2005-01-01

    control genes was used to correct five genes of interest. These genes were three genes involved in iron acquisition (tbpA, exbB, and fhuD), the heat shock protein gene groEL, and a putative quorum-sensing gene (luxS). The level of tbpA, exbB, and fhuD expression in A. pleuropneumoniae showed significant......The aims of the present investigation were to develop and test a sensitive and reproducible method for the study of gene expression in the porcine lung pathogen Actinobacillus pleuropneumoniae by real-time quantitative reverse transcription (RT)-PCR and to evaluate a number of suitable internal...... controls, as such controls have not been defined yet for this bacterium. Bacterial gene expression was studied during in vitro exponential and early stationary growth in medium with and without sufficient iron, respectively. First, the stability of expression of five genes, the glyA, tpiA, pykA, rec...

  15. Development of two real-time polymerase chain reaction assays to detect Actinobacillus pleuropneumoniae serovars 1-9-11 and serovar 2.

    Science.gov (United States)

    Marois-Créhan, Corinne; Lacouture, Sonia; Jacques, Mario; Fittipaldi, Nahuel; Kobisch, Marylène; Gottschalk, Marcelo

    2014-01-01

    Two real-time, or quantitative, polymerase chain reaction (qPCR) assays were developed to detect Actinobacillus pleuropneumoniae serovars 1-9-11 (highly related serovars with similar virulence potential) and serovar 2, respectively. The specificity of these assays was verified on a collection of 294 strains, which included all 16 reference A. pleuropneumoniae strains (including serovars 5a and 5b), 263 A. pleuropneumoniae field strains isolated between 1992 and 2009 in different countries, and 15 bacterial strains other than A. pleuropneumoniae. The detection levels of both qPCR tests were evaluated using 10-fold dilutions of chromosomal DNA from reference strains of A. pleuropneumoniae serovars 1 and 2, and the detection limit for both assays was 50 fg per assay. The analytical sensitivities of the qPCR tests were also estimated by using pure cultures and tonsils experimentally spiked with A. pleuropneumoniae. The detection threshold was 2.5 × 10(4) colony forming units (CFU)/ml and 2.9 × 10(5) CFU/0.1 g of tonsil, respectively, for both assays. These specific and sensitive tests can be used for the serotyping of A. pleuropneumoniae in diagnostic laboratories to control porcine pleuropneumonia.

  16. Induction of protective immune responses against the challenge of Actinobacillus pleuropneumoniae by the oral administration of transgenic tobacco plant expressing ApxIIA toxin from the bacteria.

    Science.gov (United States)

    Lee, Kyung-Yeol; Kim, Dong-Heon; Kang, Tae-Jin; Kim, Ju; Chung, Gook-Hyun; Yoo, Han-Sang; Arntzen, Charles J; Yang, Moon-Sik; Jang, Yong-Suk

    2006-12-01

    Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia. Among the virulence factors, ApxIIA, a bacterial exotoxin, is reportedly expressed in many serotypes and is considered as a candidate for the development of a vaccine against the bacterial infection. Previously, we isolated a field strain of A. pleuropneumoniae serotype 2 in Korea and characterized its exotoxins to develop an oral vaccine. In this study, we initially confirmed the immunogenicity of ApxIIA expressed in Escherichia coli. We then developed transgenic tobacco expressing ApxIIA and tested its efficacy to induce a protective immune response against A. pleuropneumoniae infection after oral administration of the plant powder. We observed that protective immune responses were induced in mice after oral administration of the plant powder once a week for 4 weeks. Immunoassays revealed that the levels of antigen-specific immunoglobulin G against ApxIIA increased in mice that were fed a powder made from the transgenic plant, but not in mice fed a powder made from wild-type tobacco. Additionally, mice fed the transgenic plant powder were protected from an injection of a lethal dose of A. pleuropneumoniae. These results support that the transgenic plant may be a suitable candidate for an oral vaccine that could be used effectively against A. pleuropneumoniae infection.

  17. Pharmacokinetic/pharmacodynamic evaluation of marbofloxacin in the treatment of Haemophilus parasuis and Actinobacillus pleuropneumoniae infections in nursery and fattener pigs using Monte Carlo simulations.

    Science.gov (United States)

    Vilalta, C; Giboin, H; Schneider, M; El Garch, F; Fraile, L

    2014-12-01

    This study evaluated the theoretical clinical outcome of three marbofloxacin posology regimens in two groups of pigs (weaners and fatteners) for the treatment of Actinobacillus pleuropneumoniae (App) and Haemophilus parasuis (Hp) infection and the appearance of resistant bacteria due to the antibiotic treatment. The probability of target attainment (PTA) for pharmacokinetic/pharmacodynamics (PK/PD) ratios associated with clinical efficacy and with the appearance of antimicrobial resistance for fluoroquinolones at each minimum inhibitory concentration (MIC) or mutant prevention concentration (MPC) were calculated, respectively. The cumulative fraction of response (CFR) was calculated for the three posology regimens against App and they ranged from 91.12% to 96.37% in weaners and from 93% to 97.43% in fatteners, respectively. In the case of Hp, they ranged from 80.52% to 85.14% in weaners and from 82.01% to 88.49% in fatteners, respectively. Regarding the PTA of the PK/PD threshold associated with the appearance of antimicrobial resistance, results showed that marbofloxacin would prevent resistances in most of the animals up to the MPC value of 1 μg/mL.

  18. Mitogen-activated protein kinases p38 and JNK mediate Actinobacillus pleuropneumoniae exotoxin ApxI-induced apoptosis in porcine alveolar macrophages.

    Science.gov (United States)

    Wu, Chi-Ming; Chen, Zeng-Weng; Chen, Ter-Hsin; Liao, Jiunn-Wang; Lin, Cheng-Chung; Chien, Maw-Sheng; Lee, Wei-Cheng; Hsuan, Shih-Ling

    2011-08-05

    Actinobacillus pleuropneumoniae exotoxins (Apx) are major virulence factors that play important roles in the pathogenesis of pleuropneumonia in swine. A previous study has demonstrated that native ApxI at low concentrations induces apoptosis in primary porcine alveolar macrophages (PAMs) via a caspase-3-dependent pathway. However, the molecular mechanisms underlying ApxI-induced apoptosis remain largely unknown. In this study, it was shown that ApxI treatment in PAMs rapidly induced phosphorylation of both p38 and JNK, members of the mitogen-activated protein kinase family. Application of a selective p38 or JNK inhibitor significantly reduced ApxI-induced apoptosis, indicating the involvement of p38 and JNK pathways in this event. Furthermore, activation of both caspase-8 and -9 were observed in ApxI-stimulated PAMs. Inhibition of caspase-8 and caspase-9 activity significantly protected PAMs from ApxI-induced apoptosis. In addition, Bid activation was also noted in ApxI-treated PAMs, and inhibition of caspase-8 suppressed the activation of Bid and caspase-9, suggesting that ApxI was able to activate the caspases-8-Bid-caspase-9 pathway. Notably, inhibition of p38 or JNK pathway greatly attenuated the activation of caspases-3, -8, and -9. This study is the first to demonstrate that ApxI-induced apoptosis of PAMs involves the activation of p38 and JNK, and engages the extrinsic and intrinsic apoptotic pathways.

  19. Adh enhances Actinobacillus pleuropneumoniae pathogenicity by binding to OR5M11 and activating p38 which induces apoptosis of PAMs and IL-8 release.

    Science.gov (United States)

    Wang, Lei; Qin, Wanhai; Zhang, Jing; Bao, Chuntong; Zhang, Hu; Che, Yanyi; Sun, Changjiang; Gu, Jingmin; Feng, Xin; Du, Chongtao; Han, Wenyu; Richard, Paul Langford; Lei, Liancheng

    2016-04-05

    Members of the Trimeric Autotransporter Adhesin (TAA) family play a crucial role in the adhesion of Gram-negative pathogens to host cells, but the immunopathogenesis of TAAs remains unknown. Our previous studies demonstrated that Adh from Actinobacillus pleuropneumoniae (A. pleuropneumoniae) is required for full bacterial pathogenicity. Alveolar macrophages are the first line of defense against respiratory infections. This study compared the interactions between porcine alveolar macrophages (PAMs) and wild-type A. pleuropneumoniae (5b WT) or an Adh-deletion strain (5b ΔAdh) via gene microarray, immunoprecipitation and other technologies. We found that Adh was shown to interact with the PAMs membrane protein OR5M11, an olfactory receptor, resulting in the high-level secretion of IL-8 by activation of p38 MAPK signaling pathway. Subsequently, PAMs apoptosis via the activation of the Fax and Bax signaling pathways was observed, followed by activation of caspases 8, 9, and 3. The immunological pathogenic roles of Adh were also confirmed in both murine and piglets infectious models in vivo. These results identify a novel immunological strategy for TAAs to boost the pathogenicity of A. pleuropneumoniae. Together, these datas reveal the high versatility of the Adh protein as a virulence factor and provide novel insight into the immunological pathogenic role of TAAs.

  20. Identification and Detection of Actinobacillus pleuropneumoniae in Infected and Subclinically Infected Pigs by Multiplex PCR Based on the Genes ApxIVA and OmlA

    Institute of Scientific and Technical Information of China (English)

    XIAO Guo-sheng; CAO San-jie; DUAN Li-li; WEN Xin-tian; MA Xiao-ping; CHEN Hua-mei

    2006-01-01

    PCRs based on different genes of Actinobacillus pleuropneumoniae have been developed for detecting and identifying A. pleuropneumoniae. Some of them could amplify positive fragments from the phylogenetically closely related species bacteria. To improve veracity and specificity of PCR, a species-specific multiplex PCR assay was developed to identify and detect A. pleuropneumoniae, based on the 3'-terminus of the species-specific apxIVA gene and the already existing species-specific primers in the omlA gene. Both 346-bp and 950-bp fragments could be simultaneously amplified from all A. pleuropneumoniae reference strains and isolates, and the species specificity of the assay was evaluated with a collection of ten strains representing eight different species bacteria including species normally found in the respiratory tracts of swine. All of these strains turned out negative in the multiplex PCR. All sequences of products of multiplex PCR randomly sampled were also correct. The sensitivity of the multiplex PCR was determined to be 10 pg ofA. pleuropneumoniae DNA. The multiplex PCR and bacterial isolation were compared to determine their sensitivities by using experimentally infected pigs and clinical disease pigs. The multiplex PCR was more sensitive than bacterial isolation. The multiplex PCR was also evaluated on mixed bacterial cultures from clinical healthy pigs. 26/100 (26%) of the subclinically infected pigs were detected from clinical healthy pigs. The results indicate that the multiplex PCR assay is a sensitive, highly specific,and effective diagnostic tool for identification and detection of A. pleuropneumoniae.

  1. Magnetic beads-based enzymatic spectrofluorometric assay for rapid and sensitive detection of antibody against ApxIVA of Actinobacillus pleuropneumoniae.

    Science.gov (United States)

    Wei, Bo; Li, Fang; Yang, Huicui; Yu, Lei; Zhao, Kaihong; Zhou, Rui; Hu, Yonggang

    2012-05-15

    In this paper, a simple, easily-operated and enzyme-amplified fluorescence immunoassay method using magnetic particles for the detection of antibody against Actinobacillus pleuropneumoniae (APP) has been presented. The A protein of APP Repeats-in-Toxin IV (ApxIVA) with high specificity to the APP species was immobilized onto the magnetic bead surfaces. Horseradish peroxidase (HRP), which can catalyze the substrate 4-hydroxyphenylacetic acid (p-HPA), generating fluorescent bi-p, p'-hydroxyphenylacetic acid (DBDA), was selected as an enzymatic-amplified tracer. The ApxIVA antibody was detected for the presence of APP infection by measuring the fluorescence intensity of DBDA. Under optimal conditions, the calibration plot obtained for standard positive serum was approximately linear within the dilution range 1:160-1:5120. The limit of detection (LOD) for the assay was 1:10240, considerably lower than that of ApxIVA-ELISA (1:320) (S/N=3). A series of repeatability measurements of using 1:320-fold diluted standard positive serum gave reproducible results with a relative standard deviation (RSD) of 4.8% (n=11). The ability of the immunosensor to analyze clinical samples was tested on porcine sera. The immunosensor yielded an efficiency of 89.7%, sensitivity of 90.9% and specificity of 89.3% compared with ApxIVA-ELISA.

  2. One of two TolC-like proteins is involved in antibiotic resistance and biofilm formation of Actinobacillus pleuropneumoniae clinical isolate SC1516

    Directory of Open Access Journals (Sweden)

    Ying Li

    2016-10-01

    Full Text Available Actinobacillus pleuropneumoniae is the etiologic agent of porcine contagious pleuropneumonia, a significant disease that causes serious economic losses to the swine industry worldwide. Persistent infections caused by bacterial biofilms are recalcitrant to treat because of the particular drug resistance of biofilm-dwelling cells. TolC, a key component of multidrug efflux pumps, are responsible for multidrug resistance in many Gram-negative bacteria. In this study, we identified two TolC-like proteins, TolC1 and TolC2, in A. pleuropneumoniae. Deletion of tolC1, but not tolC2, caused a significant reduction in biofilm formation, as well as increased drug sensitivity of both planktonic and biofilm cells. The genetic-complementation of the tolC1 mutation restored the competent biofilm and drug resistance. Besides, biofilm formation was inhibited and drug sensitivity was increased by the addition of phenylalanine-arginine beta-naphthylamide (PAβN, a well-known efflux pump inhibitor (EPI, suggesting a role for EPI in antibacterial strategies towards drug tolerance of A. pleuropneumoniae. Taken together, TolC1 is required for biofilm formation and is a part of the multidrug resistance machinery of both planktonic and biofilm cells, which could supplement therapeutic strategies for resistant bacteria and biofilm-related infections of A. pleuropneumoniae clinical isolate SC1516.

  3. Identificación de Actinobacillus pleuropneumoniae biotipo 1, serotipo 1, de pulmones de cerdo con y sin lesiones neumónicas utilizando la técnica de inmunohistoquímica

    OpenAIRE

    Rigoberto Hernández Castro; Gilberto Chávez Gris; José Ángel Gutiérrez Pabello

    2002-01-01

    En el presente estudio se comparó el aislamiento bacteriológico y una técnica de inmunohistoquímica con la variedad del complejo avidina-biotina-peroxidasa para la identificación de Actinobacillus pleuropneumnoniae biotipo 1, serotipo 1. Se utilizaron 100 pulmones de cerdos clínicamente sanos sacrificados en rastro, 50 con lesiones macroscópicas aparentes y 50 sin lesiones macroscópicas. Todas las muestras se analizaron mediante histopatología, bacteriología e inmunohistoquímica. Las lesiones...

  4. Estimation of sensitivity, specificity and predictive values of two serologic tests for the detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in the absence of a reference test (gold standard)

    DEFF Research Database (Denmark)

    Enøe, Claes; Andersen, Søren; Sørensen, Vibeke

    2001-01-01

    independence was assumed to models allowing for conditional dependence, given the true disease status. No strong evidence of conditional dependence in either test sensitivity or specificity was found. Assuming independence, maximum-likelihood estimates and 95% confidence intervals of the sensitivity...... Actinobacillus pleuropneumoniae serotype 2 in a survey of respiratory diseases in Danish finishing pigs. The estimates were obtained by maximum-likelihood and also by a Bayesian method (implemented with Gibbs sampling). Possible dependence of diagnostic errors was investigated by comparing models where...

  5. Actinobacillus pleuropneumoniae is impaired by the garlic volatile allyl methyl sulfide (AMS) in vitro and in-feed garlic alleviates pleuropneumonia in a pig model.

    Science.gov (United States)

    Becker, Petra M; van Wikselaar, Piet G; Mul, Monique F; Pol, Arjan; Engel, Bas; Wijdenes, Jan W; van der Peet-Schwering, Carola M C; Wisselink, Henk J; Stockhofe-Zurwieden, Norbert

    2012-01-27

    Decomposition products of ingested garlic are to a certain extent excreted via the lungs. If the supposed health-supporting capacities associated with garlic extend to these exhaled sulfurous compounds, they could have an effect on the course of pneumonia. In this study, the garlic-derived volatile allyl methyl sulfide (AMS) as a lead compound of volatile garlic metabolites was shown to exhibit an antibacterial effect against the pig pathogen Actinobacillus pleuropneumoniae serotype 9. AMS caused a delay in the appearance of the optical density-monitored growth of A. pleuropneumoniae in medium when compared to unaffected growth curves, yet without lowering the stationary phase yield at the concentration range tested. At 1.1mM, AMS impaired the in vitro growth rate of A. pleuropneumoniae serotype 9 by 8% compared to unimpeded growth. In an animal trial, a garlic-fed group of 15 pigs that received a diet with 5% garlic feed component and a control group of 15 pigs that received a diet without garlic were infected with A. pleuropneumoniae serotype 2 via an aerosol and subsequently followed for 4 days. At the day of the challenge, blood AMS in the garlic-fed group amounted to 0.32 ± 0.13 μM. A beneficial, alleviating effect of garlic on the course and severity of an A. pleuropneumoniae infection in pigs was indicated by the reduced occurrence of characteristic pleuropneumonia lesions (27% of the lungs affected in the garlic-fed group vs. 47% in the control group) and a near to significant (p=0.06) lower relative lung weight post mortem in the garlic-fed group.

  6. Usefulness of real time PCR for the differentiation and quantification of 652 and JP2 Actinobacillus actinomycetemcomitans genotypes in dental plaque and saliva

    Directory of Open Access Journals (Sweden)

    Piras Vincenzo

    2006-06-01

    Full Text Available Abstract Background The aim of our study is to describe a fast molecular method, able to distinguish and quantize the two different genotypes (652 and JP2 of an important periodontal pathogen: Actinobacillus actinomycetemcomitans. The two genotypes show differences in the expression of an important pathogenic factor: the leukotoxin (ltx. In order to evidence this, we performed a real time PCR procedure on the ltx operon, able to recognize Aa clinical isolates with different leukotoxic potentials. Methods The specificity of the method was confirmed in subgingival plaque and saliva specimens collected from eighty-one Italian (Sardinian subjects with a mean age of 43.9, fifty five (68 % of whom had various clinical forms of periodontal disease. Results This procedure showed a good sensitivity and a high linear dynamic range of quantization (107-102 cells/ml for all genotypes and a good correlation factor (R2 = 0.97–0.98. Compared with traditional cultural methods, this real time PCR procedure is more sensitive; in fact in two subgingival plaque and two positive saliva specimens Aa was only detected with the molecular method. Conclusion A low number of Sardinian patients was found positive for Aa infections in the oral cavity, (just 10 positive periodontal cases out of 81 and two of these were also saliva positive. The highly leukotoxic JP2 strain was the most representative (60 % of the positive specimens; the samples from periodontal pockets and from saliva showed some ltx genotype for the same patient. Our experience suggests that this approach is suitable for a rapid and complete laboratory diagnosis for Aa infection.

  7. Cloning and Expression of Actinobacillus pleuropneumoniae Gene Coding for TbpA and Development of an Indirect TbpA-ELISA

    Institute of Scientific and Technical Information of China (English)

    LIANG Wang-wang; HE Qi-gai; CHEN Huan-chun; XU Di-ping; WU Rui; ZHANG Rong-rong

    2008-01-01

    This study presents the cloning and expression of gene encoding transferrin-binding protein A from Actinobacillus pleuropneumoniae in Escherichia coli expression system and the development of an indirect TbpA-ELISA. The gene coding TbpA was amplified from the A. pleuropneumoniae serotype 2 genome using polymerase chain reaction and cloned to pET-28b expression vector under the control of strong, inducible T7 promoter. The recombinant plasmid was expressed in E. coli BL21 (DE3). The expressed fusion protein was analyzed using SDS-PAGE and Western blotting. The diagnostic potential of recombinant TbpA (rTbpA) was evaluated through an antibody-detection indirect ELISA based on the purified rTbpA. The TbpA antibodies were detectable in mice on day 7 after vaccination with purified rTbpA protein or infection with A. pleuropneumoniae serotype 10 with the TbpA-based ELISA. In addition, the TbpA-ELISA was able to detect 12 serotyping rabbit antisera postinoculation (PI) with A. pleuropneumoniae 12 serotypes experimentally. The comparable result was obtained by detecting the 117 clinical serum samples using, respectively, the TbpA-ELISA and indirect hemagglutination test (IHA) based on multiplex antigen. The result indicates that the TbpA-ELISA was the more sensitive method compared with the Mix-IHA method because of its consistent presence in A. pleuropneumoniae serotypes. In conclusion, the conserved TbpA of A. pleuropneumoniae can be used for the development of a cross-serotype diagnostic method for the detection of antibodies against A. pleuropneumoniae.

  8. A novel Respiratory Health Score (RHS supports a role of acute lung damage and pig breed in the course of an Actinobacillus pleuropneumoniae infection

    Directory of Open Access Journals (Sweden)

    Gerlach Gerald F

    2009-04-01

    Full Text Available Abstract Background Bacterial lung infections are a major cause of economic losses in the pig industry; they are responsible for approximately 50% of the antibiotics used in pigs and, therefore, also present an increasing concern to consumer protection agencies. In response to this changing market we investigated the feasibility of an old approach aimed at the breeding selection of more resistant pigs. As a first step in this direction we applied a new respiratory health score system to study the susceptibility of four different pig breeding lines (German Landrace, Piétrain, Hampshire, Large White towards the respiratory tract pathogen Actinobacillus (A. pleuropneumoniae. Results A controlled experimental aerosol infection with an A. pleuropneumoniae serotype 7 isolate was performed using 106 weaning pigs of defined breeding lines from the breeds German Landrace, Piétrain, Hamphire, and Large White. Pigs were clinically assessed on days 4 and 20 post infection following a novel scoring system, the Respiratory Health Score (RHS, which combines clinical, sonographic and radiographic examination results. The ranking on day 4 was significantly correlated with the ranking based on the pathomorphological Lung Lesion Score (LLS; Spearman Rank Correlation Coefficient of 0.86 [p Conclusion These results demonstrate that the RHS obtained from live pigs shows a highly significant correlation to the lung lesion score considered as a "gold standard". The correlation of the ranking at days 4 and 20 post infection implies that the course of disease is highly dependent on the acute lung damage. The different severity of signs among the tested pig breeding lines clearly suggests a genetic difference in the susceptibility of pigs to A. pleuropneumoniae infection.

  9. Mechanisms underlying Actinobacillus pleuropneumoniae exotoxin ApxI induced expression of IL-1β, IL-8 and TNF-α in porcine alveolar macrophages

    Directory of Open Access Journals (Sweden)

    Chen Zeng-Weng

    2011-02-01

    Full Text Available Abstract Actinobacillus pleuropneumoniae (A. pleuropneumoniae causes fibrino-hemorrhagic necrotizing pleuropneumonia in pigs. Production of proinflammatory mediators in the lungs is an important feature of A. pleuropneumoniae infection. However, bacterial components other than lipopolysaccharide involved in this process remain unidentified. The goals of this study were to determine the role of A. pleuropneumoniae exotoxin ApxI in cytokine induction and to delineate the underlying mechanisms. Using real-time quantitative PCR analysis, we found native ApxI stimulated porcine alveolar macrophages (PAMs to transcribe mRNAs of IL-1β, IL-8 and TNF-α in a concentration- and time-dependent manner. Heat-inactivation or pre-incubation of ApxI with a neutralizing antiserum attenuated ApxI bioactivity to induce cytokine gene expression. The secretion of IL-1β, IL-8 and TNF-α protein from PAMs stimulated with ApxI was also confirmed by quantitative ELISA. In delineating the underlying signaling pathways contributing to cytokine expression, we observed mitogen-activated protein kinases (MAPKs p38 and cJun NH2-terminal kinase (JNK were activated upon ApxI stimulation. Administration of an inhibitor specific to p38 or JNK resulted in varying degrees of attenuation on ApxI-induced cytokine expression, suggesting the differential regulatory roles of p38 and JNK in IL-1β, IL-8 and TNF-α production. Further, pre-incubation of PAMs with a CD18-blocking antibody prior to ApxI stimulation significantly reduced the activation of p38 and JNK, and subsequent expression of IL-1β, IL-8 or TNF-α gene, indicating a pivotal role of β2 integrins in the ApxI-mediated effect. Collectively, this study demonstrated ApxI induces gene expression of IL-1β, IL-8 and TNF-α in PAMs that involves β2 integrins and downstream MAPKs.

  10. Distribution of lymphocytes, immunoglobulin-containing cells, macrophages, and dendritic cells in the accessory sex glands of rams experimentally infected with Actinobacillus seminis

    Directory of Open Access Journals (Sweden)

    Jorge Acosta-Dibarrat

    2016-05-01

    Full Text Available Abstract: The distribution of cells involved in the immune response in accessory sex glands of rams experimentally infected with Actinobacillus seminis was studied. Twelve one-year old rams were experimentally infected by intraurethral (IU (n=4 and intraepididymal (IE (n=4 route, and four control (CON animals were used. The animals were slaughtered 35 days post-inoculation, samples were taken from accessory sex glands, and bacteriology and histopathology tests were performed. The presence of CD4, CD8 and TCRγδ (WC1 lymphocytes, CD45RO cells, macrophages (CD14, dendritic cells (CD1b, IgA-, IgG- and IgM-containing cells (IgCC was determined. Animals of the IE group developed clinical epididymitis. No lesions were seen in rams of the IU group; two of the intraepididymal inoculated CON developed small lesions in the epididymis. A. seminis isolates were achieved from 6:16 (37.5% accessory sex glands in the IE group, but not in the IU and CON groups. In the CON group, IgA- and IgM- containing cells predominated in the bulbourethral glands and the disseminated prostate, and they were scarce or null in the vesicles and ampullae. A significant increase of IgA-, IgG- and IgM- containing cells was confirmed in the seminal vesicles, the ampullae and the bulbourethral glands in the IE group. In the IE and IU groups, an increase in CD4, CD8, WC1, CD45RO and CD14 was evidenced in the vesicles and ampullae. CD1b dendritic cells were present in the ampullae and vesicles with inflammatory processes. A. seminis triggered a local immune response in the IE and IU groups. These results indicate a different pattern of infiltrating immune cells in the accessory sex glands of infected A. seminis rams.

  11. Iron acquisition in the dental pathogen Actinobacillus actinomycetemcomitans: what does it use as a source and how does it get this essential metal?

    Science.gov (United States)

    Rhodes, Eric R; Menke, Sharon; Shoemaker, Christopher; Tomaras, Andrew P; McGillivary, Glen; Actis, Luis A

    2007-06-01

    Actinobacillus actinomycetemcomitans requires iron to grow under limiting conditions imposed by synthetic and natural chelators. Although none of the strains tested used hemoglobin, lactoferrin or transferrin, all of them used FeCl3 and hemin as iron sources under chelated conditions. Dot-blot binding assays showed that all strains bind lactoferrin, hemoglobin, and hemin but not transferrin. When compared with smooth strains, the rough isolates showed higher hemin binding activity, which was sensitive to proteinase K treatment. A. actinomycetemcomitans harbors the Fur-regulated afeABCD locus coding for iron acquisition in isogenic and non-isogenic cell backgrounds. The genome of this oral pathogen also harbors several other predicted iron uptake genes including the hitABC locus, which restored iron acquisition in the E. coli 1017 ent mutant. However, the disruption of this locus in the parental strain did not affect iron acquisition as drastically as the inactivation of AfeABCD, suggesting that the latter system could be more involved in iron transport than the HitABC system. The genome of this oral pathogen also harbors an active copy of the exbBexbDtonB operon, which could provide the energy needed for hemin acquisition. However, inactivation of each coding region of this operon did not affect the hemin and iron acquisition phenotypes of isogenic derivatives. This observation suggests that the function of these proteins could be replaced by those coded for by tolQ, tolR and tolA as it was described for other bacterial transport systems. Interruption of a hasR homolog, an actively transcribed gene that is predicted to code for an outer membrane hemophore receptor protein, did not affect the ability of an isogenic derivative to bind and use hemin under chelated conditions. This result also indicates that A. actinomycetemcomitans could produce more than one outer membrane hemin receptor as it was described in other human pathogens. All strains tested formed biofilms

  12. Association between infection of different strains of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in subgingival plaque and clinical parameters in chronic periodontitis

    Institute of Scientific and Technical Information of China (English)

    WU Yan-min; YAN Jie; CHEN Li-li; GU Zhi-yuan

    2007-01-01

    Objective: The aim of this study was to investigate subgingival infection frequencies ofPorphyromonas gtngivalis and Actinobacillus actinomycetemcomitans strains with genetic variation in Chinese chronic periodontitis (CP) patients and to evaluate its correlation with clinical parameters. Methods: Two multiplex polymerase chain reaction (PCR) assays were developed to detect the 16SrDNA, collagenase (prtC) and fimbria (fimA) genes of P. gingivalis and the 16SrDNA, leukotoxin (lktA) and fimbria-associated protein (lap) genes ofA. actinomycetemcomitans in 60 sulcus samples from 30 periodontal healthy subjects and in 122 subgingival plaque samples from 61 patients with CP. The PCR products were further T-A cloned and sent for nucleotide sequence analysis. Results: The 16SrDNA, prtC andfimA genes ofP. gingivalis were detected in 92.6%, 85.2% and 80.3% of the subgingival plaque samples respectively, while the 16SrDNA, lktA andfap genes ofA. actinomycetemcomitans were in 84.4%,75.4% and 50.0% respectively. Nucleotide sequence analysis showed 98.62%~100% homology of the PCR products in these genes with the reported sequences. P. gingivalis strains with prtC+/fimA+ and A. actinomycetemcomitans with lktA+ were predominant in deep pockets (>6 mm) or in sites with attachment loss ≥5 mm than in shallow pockets (3~4 mm) or in sites with attachment loss ≤2 mm (P<0.05). P. gingivalis strains with prtC+/fimA+ also showed higher frequency in gingival index (GI)=3than in GI= 1 group (P<0.05). Conclusion: Infection of P. gingivalis with prtC+/fimA+ and A. actinomycetemcomitans with lktA+correlates with periodontal destruction of CP in Chinese. Nonetheless P. gingivalis fimA, prtC genes and A. actinomycetemcomitans IktA gene are closely associated with periodontal destruction, while A. actinomycetemcomitansfap gene is not.

  13. Research advancement on Actinobacillus actinomycetemcomitans cytolethal distending toxin%伴放线放线杆菌细胞致死性扩张毒素研究进展

    Institute of Scientific and Technical Information of China (English)

    段君兰

    2014-01-01

    细胞致死性扩张毒素(cytolethal distending toxin,CDT)是近年来新发现的一种伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)的毒力因子(Aa CDT),研究发现该毒力因子在Aa的牙周致病机制中发挥着重要的作用.本文就Aa CDT的结构和功能、作用机制及致病机制等方面的研究进展作一综述,以期进一步了解Aa的致病机理.

  14. Reação em Cadeia da Polimerase (PCR baseada no gene cpx para detecção de Actinobacillus pleuropneumoniae em suínos natural e experimentalmente infectados Polymerase Chain Reaction (PCR based on the cpx gene for detection of Actinobacillus pleuropneumoniae in natural and experimentally infected pigs

    Directory of Open Access Journals (Sweden)

    Karina Koerich de Souza

    2008-10-01

    Full Text Available A pleuropneumonia suína é uma das mais importantes doenças respiratórias dos suínos, estando presente em todos os países produtores. Para o controle e o monitoramento da pleuropneumonia, é necessário o desenvolvimento de métodos rápidos e acurados de diagnóstico. Com o objetivo de validar a técnica da PCR, baseada no gene cpx de Actinobacillus pleuropneumoniae, em suínos sabidamente positivos, primeiramente foi realizada inoculação experimental com amostras de A. pleuropneumoniae sorotipo 5B e coletadas amostras por meio de suabe de tonsila, biópsia de tonsila e sangue para realização da técnica de PCR, isolamento bacteriológico e teste de ELISA, respectivamente. Posteriormente, estas técnicas foram aplicadas em suínos naturalmente infectados, em três rebanhos com diferentes situações sanitárias quanto à apresentação clínica da doença. De cada rebanho, foram analisados cinco grupos de suínos com idades diferentes, sendo coletado de cada animal biópsia de tonsila para isolamento bacteriológico e PCR e sangue para determinação do perfil sorológico. Os resultados obtidos na inoculação experimental confirmaram que, mesmo com o estabelecimento da infecção comprovada pelo isolamento bacteriológico, após o período de 45 dias, não foi possível detectar o agente pela técnica de PCR. Em animais naturalmente infectados, a técnica de PCR apresentou maior sensibilidade quando comparado com o isolamento. A associação entre PCR e ELISA demonstrou ser uma boa alternativa para definir a situação sanitária do rebanho quanto à infecção por A. pleuropneumoniae.Swine pleuropneumonia is one of the most important pig respiratory diseases and has been found in all producer countries. For control and monitoring of pleuropneumonia, it is necessary the development of fast and specific methods of diagnosis. To validate PCR based on the cpx gene of Actinobacillus pleuropneumoniae in positive pigs, an experimental

  15. . and Aggregatibacter segnis comb. nov., and emended description of Aggregatibacter aphrophilus to include V factor-dependent and V factor-independent isolates, Reclassification of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Haemophilus paraphrophilus and Haemophilus segnis as Aggregatibacter actinomycetemcomitans gen. nvo., comb. nov., Aggregatibacter aphrophilus comb. nov

    DEFF Research Database (Denmark)

    Nørskov-Lauritsen, N.; Kilian, Mogens

    2006-01-01

    The aim of this study was to reinvestigate the relationships and the generic affiliations of the species Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Haemophilus paraphrophilus and Haemophilus segnis. The nicotinamide phosphoribosyltransferase gene (nadV) conferring V factor-ind......)=NCTC 5906(T)) and Aggregatibacter segnis comb. nov. (type strain HK316(T)=ATCC 33393(T)=CCUG 10787(T)=CCUG 12838(T)=CIP 103292(T)=NCTC 10977(T)). The species of the genus Aggregatibacter are independent of X factor and variably dependent on V factor for growth in vitro......The aim of this study was to reinvestigate the relationships and the generic affiliations of the species Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Haemophilus paraphrophilus and Haemophilus segnis. The nicotinamide phosphoribosyltransferase gene (nadV) conferring V factor......-independent growth was identified in Haemophilus aphrophilus. The gene encodes a polypeptide of 462 amino acids that shows 74.5 % amino acid sequence identity to the corresponding enzyme from Actinobacillus actinomycetemcomitans. Ten isolates of Haemophilus paraphrophilus all carried a nadV pseudogene. DNA from...

  16. Estudios hematológicos y patológicos comparativos de cerdos inoculados con un aislado de campo y el serotipo 5 ATCC de Actinobacillus pleuropneumoniae Comparative hematological and pathological study of inoculated pigs with a field isolate and an ATCC serotype 5 of Actinobacillus pleuropneumoniae

    Directory of Open Access Journals (Sweden)

    D Muñoz

    2010-01-01

    Full Text Available Se realizó una inoculación experimental de A. pleuropneumoniae utilizando un aislado de campo y una cepa de referencia ATCC serotipo 5, para lo cual se utilizaron tres grupos de animales (n = 15 para cada grupo. El grupo 1 (G1 fue inoculado con medio estéril, el grupo (G2 con serotipo 5 ATCC y el grupo 3 (G3 fue inoculado con un aislado de campo (418/07. Los resultados mostraron diferencias significativas (P ≤ 0,05 en el recuento de leucocitos totales entre el grupo G1 v/s G2 y G1 v/s G3 y los grados de las lesiones pulmonares totales evidenciaron diferencias estadísticamente significativas (P ≤ 0,05 entre los tres grupos de estudio. Las lesiones histopatológicas pulmonares mostraron diferencias estadísticas relevantes sólo entre G1 y G3 (P ≤ 0,05. En este trabajo se verifican diferencias importantes del comportamiento entre el aislado de campo y el serotipo 5 ATCC, sobre los cambios hematológicos y las lesiones macroscópicas e histopatológicas ocasionadas por ellos, lo cual podría indicar una mayor virulencia y patogenicidad del aislado nacional. Se espera en un futuro próximo serotipificar este aislado nacional de App.An experimental inoculation of Actinobacillus pleuropneumoniae (App was carried out with a field isolate and an ATCC serotype 5. Three groups of 15 pigs each were used. Group 1 (G1 was the control group inoculated with sterile media, Group 2 was inoculated with the serotype 5 ATCC, and Group 3 (G3 was inoculated with a field isolate (418/07. The results showed statistically significant differences (P ≤ 0.05 in the total leukocytes count between G1 v/s G2 and G1 v/s G3. The total macroscopic lung lesions scores were statistically different among the 3 groups (P ≤ 0.05. However, statistical difference was found only between G1 and G3 in the histopathological lung lesions (P ≤ 0.05. This work shows a clear difference in the hematological changes and the macroscopic and histopathological lesions between the

  17. Occurrence of Actinobacillus actinomycetemcomitans in patients with chronic periodontitis, aggressive periodontitis, healthy subjects and children with gingivitis in two cities of the state of São Paulo, Brazil Ocorrência de Actinobacillus actinomycetemcomitans em pacientes com periodontite crônica, periodontite agressiva, pessoas saudáveis e crianças com gengivite em duas cidades do Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Elerson Gaetti Jardim Júnior

    2006-06-01

    Full Text Available The aim of this study was to determine the frequency of isolation of Actinobacillus actinomycetemcomitans (Aa in 100 patients with chronic periodontitis, 14 patients with aggressive periodontitis, 142 pre-school children with gingivitis and 134 periodontally healthy subjects. Samples of subgingival plaque were taken using sterilized paper points introduced into periodontal pockets or gingival crevice for 60 seconds and inoculated on TSBV agar, which was incubated under anaerobiosis at 37ºC, for 4 days. Microbial identification was performed through biochemical methods and morphocellular and morphocolonial analysis. Aa was detected in 40.3% of healthy subjects, 68% of patients with chronic periodontitis, 92.86% of patients with aggressive periodontitis and 40.14% of children with gingivitis. The rate of recovery of Aa in the tested human groups proved to be higher than previously reported and in agreement with participation of this facultative anaerobe as a member of native microbiota of the periodontium and its relation with aggressive and chronic periodontitis in Brazil.Avaliou-se a ocorrência de Actinobacillus actinmycetemcomitans (Aa em pacientes 100 pacientes com periodontite crônica, 14 com doença periodontal agressiva, 142 crianças com gengivite em idade pré-escolar e 134 indivíduos adultos saudáveis. Amostras de placa subgengival foram coletadas usando cones de papel estéreis introduzidos nas bolsas periodontais ou no sulco gengival por 60 segundos e inoculadas em ágar TSBV, que foram incubadas em anaerobiose a 37ºC, por 4 dias. A identificação microbiana foi realizada através de análises bioquímicas, morfocelulares e morfocoloniais. Aa foi detectado em 40,3% de indivíduos saudáveis, 68% de pacientes com periodontite crônica, 92,86% de pacientes com periodontite agressiva e 40,14% das crianças com gengivite. A taxa de ocorrência de Aa nos grupos testados provou ser mais alta do que a previamente descrita na literatura

  18. 耐酸性高产琥珀酸放线杆菌的诱变选育%Mutagenesis breeding of Actinobacillus succinogenes for aciduric and high-producing

    Institute of Scientific and Technical Information of China (English)

    王丹; 张静; 张晶; 王玉华

    2012-01-01

    在确定产琥珀酸放线杆菌ATCC55618菌株生长的临界pH为4.5之后,对其进行紫外诱变,筛选出耐pH3.5强酸的菌株M1,产量为17.25g/L,较原始菌株提高了12.09%,之后用含0.1%的溴甲酚绿变色平板筛选,对M1进行两轮紫外-亚硝基胍复合诱变,筛选出琥珀酸高产菌株R2,该菌株琥珀酸产量为27.35g/L,较原始菌株产量提高了77.71%。%Determine the borderline pH 4.5 of Actinobacillus succinogenes ATCC 55618, a mutant strain M1 that tolerance low pH 3.5 was obtained by mutagenesis ultraviolet radiation (UV), whose yield is 17.25 g/L, improved 12.09% than original strain, and then it was irradiated by both UV and nitrosoguanidine (NTG), screened by plate that contains 0.1% bromine cresol green (BCG), After two wheels of the compound mutation, a high producing strain R2 was obtained, whose yield of succinic acid is 27.35 g/L, improved 77.71% than the original strain ATCC 55618.

  19. Efficacy of Marbofloxacin against Experimentally Induced Actinobacillus pleuropneumoniae in Swine%麻保沙星对实验性猪传染性胸膜肺炎的药效学研究

    Institute of Scientific and Technical Information of China (English)

    邹明; 曾振灵

    2012-01-01

    采用二倍稀释法测定了麻保沙星等对猪胸膜肺炎放线杆菌的体外抑菌作用,然后对人工感染胸膜肺炎放线杆菌的猪进行临床治疗试验.猪人工发病4h后,分别以1.25、2.5、5 mg/kg体重的剂量肌注给药麻保沙星(每组10头),1 d 1次,连续4 d.结果表明:麻保沙星对胸膜肺炎放线杆菌的最小抑菌浓度为0.01 μg/mL;对猪传染性胸膜肺炎,麻保沙星(2.5、5 mg/kg)有显著疗效,治愈率分别为80%及90%.%The efficacy of marbofloxacin against experimentally induced Actinobacillus pleuropneumoniae in swine was tested to provide the experimental basis for its broad clinical application. 4 h later after the artificial inoculation infection, the swine were treated with the dosage of 1.25, 2.5, 5 mg/kg body weight once daily by intramuscular administration for 4 successive days. The results showed that in vitro minimal inhibitory concentration ( MIC) of marbofloxacin against Actinobacillius pleuropneumoniae was 0. 01 μg/mL. The therapeutic trials showed that marbofloxacin(2.5, 5 mg/kg) was efficacious in the control of A. pleuropneumoniae infection in swine, and the curative rates were 80 % and 90 % , respectively.

  20. Differences in purinergic amplification of osmotic cell lysis by the pore-forming RTX toxins Bordetella pertussis CyaA and Actinobacillus pleuropneumoniae ApxIA: the role of pore size.

    Science.gov (United States)

    Masin, Jiri; Fiser, Radovan; Linhartova, Irena; Osicka, Radim; Bumba, Ladislav; Hewlett, Erik L; Benz, Roland; Sebo, Peter

    2013-12-01

    A large subgroup of the repeat in toxin (RTX) family of leukotoxins of Gram-negative pathogens consists of pore-forming hemolysins. These can permeabilize mammalian erythrocytes (RBCs) and provoke their colloid osmotic lysis (hemolytic activity). Recently, ATP leakage through pannexin channels and P2X receptor-mediated opening of cellular calcium and potassium channels were implicated in cell permeabilization by pore-forming toxins. In the study described here, we examined the role played by purinergic signaling in the cytolytic action of two RTX toxins that form pores of different sizes. The cytolytic potency of ApxIA hemolysin of Actinobacillus pleuropneumoniae, which forms pores about 2.4 nm wide, was clearly reduced in the presence of P2X7 receptor antagonists or an ATP scavenger, such as pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), Brilliant Blue G, ATP oxidized sodium salt, or hexokinase. In contrast, antagonists of purinergic signaling had no impact on the hemolytic potency of the adenylate cyclase toxin-hemolysin (CyaA) of Bordetella pertussis, which forms pores of 0.6 to 0.8 nm in diameter. Moreover, the conductance of pores formed by ApxIA increased with the toxin concentration, while the conductance of the CyaA single pore units was constant at various toxin concentrations. However, the P2X7 receptor antagonist PPADS inhibited in a concentration-dependent manner the exacerbated hemolytic activity of a CyaA-ΔN489 construct (lacking 489 N-terminal residues of CyaA), which exhibited a strongly enhanced pore-forming propensity (>20-fold) and also formed severalfold larger conductance units in planar lipid bilayers than intact CyaA. These results point to a pore size threshold of purinergic amplification involvement in cell permeabilization by pore-forming RTX toxins.

  1. Genotyping of Actinobacillus actinomycetemcomitans isolated from subgingival plaques of patients with chronic periodontitis%慢性牙周炎患者龈下菌斑中伴放线放线杆菌基因型的分析

    Institute of Scientific and Technical Information of China (English)

    陈莉丽; 吴燕岷; 严杰; 孙伟莲

    2002-01-01

    目的建立龈下菌斑标本中伴放线放线杆菌(Actinobacillus actinomycetemcomitans, Aa)PCR检测方法,了解慢性牙周炎患者不同牙位的龈下菌斑中Aa的感染率及其优势基因型. 方法 61例慢性牙周炎患者每例采取2个不同牙位共122份龈下菌斑标本,采用培养法分离Aa菌株,以PCR或多重PCR检测16S rDNA基因、lktA基因和fap基因,部分扩增产物克隆后测序. 结果在11例患者的11份龈下菌斑标本中分离到Aa菌株.122份龈下菌斑中Aa 16S rDNA、lktA和fap检测阳性率分别为84.4%、75.4%和50.0%.38.8%的患者(19/49)不同牙位龈下菌斑中检出的Aa基因型不一致.Aa有4种基因型,其优势基因型是16S rDNA+/lktA+/fap+,其次为16S rDNA+/lktA+/fap-.部分标本上述3种基因的扩增片段与文献报道核苷酸序列的同源性为93.75%~100%. 结论建立的PCR或多重PCR有较高的敏感性和特异性,适用于龈下菌斑标本中Aa的快速检测.慢性牙周炎患者Aa感染率较高,并存在优势基因型,部分患者可被不同基因型的菌株同时感染.

  2. Isolation and Identification of Porcine Infectious Actinobacillus Pleuropneumonia in Jinzhou Area%锦州地区猪传染性胸膜肺炎放线杆菌的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    隋慧

    2012-01-01

    [Objective] The research aimed to investigate the prevalence of porcine infectious pleuropneumonia in Jinzhou area. [ Method ] Five shares of diseased materials were colleted from some pig farms of Jinzhou area. The pathogens were isolated and identified by the culture test, biochemical test,satellite phenomenon examination,hematolysis test,CAMP test,drug sensitivity test and animal inoculation test. [ Result] The pathogens were Gram - negative short bacillus,with polymorphism and no spore. Five strains of porcine infectious actinobacillus pleuropneumonia were isolated and they all had satellite phenomenon and hematolysis phenomenon. CAMP test results showed that its hemolytic circle was strengthened by Staphylococcus aureus. Five isolated strains showed high sensitivity to cefradine, cephalosporin V and norfloxacin, but they were resistant to amoxycillin,streptomycin,penicillin and aureomycin. The animal inoculation test showed that the pathogen had high pathogenicity to rabbit. [Conclusion] The research could provide basis for the clinical treatment of porcine infectious pleuropneumonia.%[目的]调查锦州猪传染胸膜肺炎的流行情况.[方法]从锦州部分发病猪场采集病料5份,通过细菌的分离培养、生化试验、卫星现象观察,溶血试验、CAMP试验、药敏试验和动物接种试验对致病菌进行分离与鉴定.[结果]该病原菌为革兰氏阴性短小杆菌,无芽孢,具有多形性.分离到5株猪传染性胸膜肺炎放线杆菌,均有卫星现象和溶血现象.CAMP试验结果表明,金黄色葡萄球菌可增强其溶血圈.5株分离菌对头孢拉定、先锋V、氟哌酸高度敏感,而对阿莫西林、链霉素、青霉素、金霉素有耐药性.动物接种试验表明该致病菌对家兔具有较高的致病力.[结论]该研究可为猪传染性胸膜肺炎的临床治疗提供依据.

  3. Effect of metal ions on fermentation and metabolization of Actinobacillus Succinogenes NJ113%金属离子对产琥珀酸放线杆菌NJ113厌氧发酵代谢的影响

    Institute of Scientific and Technical Information of China (English)

    郑晓宇; 李建; 方晓江; 陈可泉; 奚永兰; 姜岷

    2011-01-01

    The effects of adding Mg2+. Mn2+, Co2+ on cell growth and succinic acid production was investigated. The metabolic flux of Actinobacillus succinogenes NJ113 was calculated. It was found that the flux of HMP increased by 445.38%, 176.23 % and 171.67% after adding 6 mmol/L Mg2+, 6 mmol/L Mn2+, 2 mmol/L Co2+ respectively, thus the reducing power was better balanced. The flux of C4 was 57.70%, 15.94% and 2.91% higher respectively, which led to the improvement of succinic acid flux by 62.69%, 18.91% and 5.01%. The key enzyme activity analysis showed that the specific activity of PEP carboxykinase (Pck) reached 568.732 U/mg, 728.049 U/mg and 339.686 U/mg with 6 mmol/L Mg2+, 6 mmol/L Mn2+, 2 mmol/L Co2+ addition respectively. As a result, the concentration of succinc acid was 27.83 g/L, 26.27 g/L, and 23.54 g/L, while the concentration of control was only 22.79 g/L.%考察了培养基中分别添加Mg2+、Mn2+、Co2+3种金属离子对Actinobacilus Succinogenes NJ113菌体生长及产酸的影响,并进行了代谢通量分析。结果表明培养基中分别添加6mmTo1/LMg2+、6mmol/LMnz+、2mmol/LCo2+后流向HMP途径的通量r17比对照组分别提高了445.38%、176.23%和171.67%,使得还原力不足的矛盾得到缓解;流向C4途径的通量r13比对照组分别提高了57.70%、15.94%和2.91%;最终使得流向丁二酸的通量r16比对照组分别提高了62.69%、18.91%和5.01%。此外,关键酶活分析结果显示分别添加Mg2+、Mn2+以及Co2+后,PEP羧化激酶(Pck)比活力由对照组的339.18U/mg分别提高到568.732U/mg、728.049U/mg和339.686U/mg。最终当培养基中分别添加6mmol/LMg2+、6immol/LMn2+、2immol/LCo2+后丁二酸产量分别为27.83 g/L、26.27 g/L和23.54 g/L,比对照的22.79 g/L分别提高22.11%、15.27%以及3.4%。

  4. 牙周炎患者唾液中伴放线放线杆菌的检出状况分析%Prevalence of Actinobacillus actinomycetemcomitans in saliva of different types of periodontitis

    Institute of Scientific and Technical Information of China (English)

    冯向辉; 张立; 孟焕新; 徐莉; 陈智滨; 释栋

    2008-01-01

    Objective To investigate the prevalence of Actinobacillus actinomyeetemcomitans (Aa) in whole saliva of different types of periodontitis and compare the detections of Aa between saliva and pooled subgingival plaque sample, and analyze the relationship between Aa and clinical conditions. Methods Unstimulated whole saliva samples and pooled subgingival samples were collected from 50 aggressive periodontitis (AgP) patients, 48 chronic periodontitis (CP) patients and 25 subjects with no periodontitis, and Aa was detected in these samples by PCR method. Results The prevalence of Aa in whole saliva of AgP patients was significantly higher than in subjects with no periodontitis (32% vs. 4%, P <0. 01) and CP patients (32% vs. 15%, P <0. 05). Aa was also more frequently detected in whole saliva sample than in pooled subgingival sample of AgP patients (32% vs. 16%, P < 0.05). Subjects younger than 30 year sold were more likely to present Aa in whole saliva (OR = 3.23, P < 0. 05) and percentage of sites with bleeding index(BI) ≥3 over 70% was a risk indicator for the presence of Aa in whole saliva. Conclusions The detection of Aa in whole saliva sample of AgP patients was more frequent than in pooled subgingival plaque samples, and also more frequent than in CP patients and subjects with no periodontitis, which suggest that Aa may participate in the initiation and progression of aggressive periodontitis.%目的 检测不同类型牙周炎患者唾液中的伴放线放线杆菌(Actinobacillusactinomycetemcomitans,Aa),探讨唾液和集合龈下菌斑中Aa检出率的差异以及唾液中Aa的存在状况与牙周临床指标的关系. 方法 收集50例侵袭性牙周炎(aggressive periodontitis,AgP)患者、48例慢性牙周炎(chronic periedontitis,CP)患者和25例非牙周炎者的非刺激性全唾液和集合龈下菌斑,应用聚合酶链反应(PcR)技术检测两种样本中的Aa. 结果 Aa在AgP患者唾液中的检出率(32%)显著高于非牙周炎者(4%)

  5. Pharmacokinetics of tildipirosin in porcine plasma, lung tissue, and bronchial fluid and effects of test conditions on in vitro activity against reference strains and field isolates of Actinobacillus pleuropneumoniae.

    Science.gov (United States)

    Rose, M; Menge, M; Bohland, C; Zschiesche, E; Wilhelm, C; Kilp, S; Metz, W; Allan, M; Röpke, R; Nürnberger, M

    2013-04-01

    The pharmacokinetics of tildipirosin (Zuprevo(®) 40 mg/mL solution for injection for pigs), a novel 16-membered-ring macrolide for the treatment for swine respiratory disease (SRD), was investigated in studies collecting blood plasma and postmortem samples of lung tissue and bronchial fluid (BF) from swine. In view of factors influencing the in vitro activity of macrolides, and for the interpretation of tildipirosin pharmacokinetics in relation to minimum inhibitory concentrations (MIC), additional experiments were conducted to study the effects of pH, carbon dioxide-enriched atmosphere, buffers, and serum on tildipirosin MICs for various reference strains and Actinobacillus (A.) pleuropneumoniae field isolates. After single intramuscular (i.m.) injection at 4 mg/kg body weight, maximum plasma concentration (Cmax) was 0.9 μg/mL observed within 23 min (Tmax ). Mean residence time from the time of dosing to the time of last measurable concentration (MRTlast) and terminal half-life (T1/2) both were about 4 days. A dose-response relationship with no significant sex effect is observed for area under the plasma concentration-time curve from time 0 to the last sampling time with a quantifiable drug concentration (AUClast) over the range of doses up to 6 mg/kg. However, linear dose proportionality could not be proven with statistical methods. The time-concentration profile of tildipirosin in BF and lung far exceeded that in blood plasma. In lung, tildipirosin concentrations reached 3.1 μg/g at 2 h, peaked at 4.3 μg/g at day 1, and slowly declined to 0.8 μg/g at day 17. In BF, tildipirosin levels were 14.3, 7.0, and 6.5 μg/g at days 5, 10, and 14. T1/2 in lung was ∼7 days. Tildipirosin is rapidly and extensively distributed to the respiratory tract followed by slow elimination. Culture media pH and carbon dioxide-enriched atmosphere (CO2 -EA) had a marked impact on in vitro activity of tildipirosin in reference strains of various rapidly growing aerobic and

  6. PCR specific for Actinobacillus pleuropneumoniae serotype 3

    DEFF Research Database (Denmark)

    Zhou, L.; Jones, S.C.P.; Angen, Øystein

    2008-01-01

    , but the method has liminations, for example, cross-reactions between serotypes 3, 6, and 8. This study describes the development of a serotype 3-specific PCR, based on the capsule locus, which can be used in a multiplex format with the organism's specific gene apxIV. The PCR test was evaluated on 266 strains...

  7. The T Cell Response to Actinobacillus actinomycetemcomitans

    Science.gov (United States)

    2006-05-01

    disease (11). Other clinical features of the disease include deep pain on mastication, first molar mobility, distolabial migration of maxillary incisors...transformed into one shot chemically competent E. coli TOP -10 (InVitrogen). 40 .tl from each transformation was grown at 37°C overnight on LB plates...5. Peripheral Blood Mononuclear Cells (PBMCs) isolation The 13 ml vacutainer with SST Gel and Clot Activator, or "Tiger Top ," was centrifuged at 3800

  8. The Comparison of Culture Characteristic and Pathogenicity of Actinobacillus pleuropneumoniae △adh with 5b%胸膜肺炎放线杆菌adh基因敲除株与5b野生株的培养特性及致病性比较

    Institute of Scientific and Technical Information of China (English)

    计群; 雷连成; 杨舒心; 翟瑞东; 张庆明; 杨峰; 韩文瑜

    2013-01-01

    三聚体自转运黏附素(trimeric autotransporter adhesin,TAA)是近年来发现的参与猪胸膜肺炎放线杆菌(Actinoba cillus pleuropneumoniae,APP)黏附宿主细胞的重要毒力因子.本研究比较了5b adh基因缺失株(△adh)与5b野生株的生物学特性及其对仔猪的致病性.结果表明,在BHI液体培养基中,△adh生长速度明显高于5b野牛株;△adh在液体培养基中细菌集聚性明显减弱;仔猪感染后临床症状典型,猪肺脏病理组织切片结果表明,△adh致病性弱于野生株.本研究结果证实adh在APP黏附宿主过程中发挥重要作用,为下一步探究APP致病机制奠定基础.%It was found that trimeric autotransporter adhesin (TAA) was the important virulence factor for Actinobacillus pleuropneumuniae(APP) adhere to host in the recent years. In this research, we compared the culture characteristic and the pathogenicity to piglet of Aadh and 5b wild strain. The results showed that the growth rate of Aadh was higher than 5b obviously; the Aadh showed weaker aggregation compared with 5b in liquid culture condition; the piglet showed typical symptoms after the infection, the observation of HE showed that pathogenicity of the Aadh was weaker than 5b wild strain. The result of the comparisons conformed the importance of adh in the progress of APP attacking to host and set basics in the pathogenic research of APP in the next step.

  9. 胸膜肺炎放线杆菌菌影疫苗免疫仔猪前后差异表达基因的鉴定与分析%Identification and analysis of differential expression genes in peripheral blood lymphocytes from piglet immunized by bacterial ghost of Actinobacillus pleuropneumoniae

    Institute of Scientific and Technical Information of China (English)

    杨舒心; 雷连成; 杜崇涛; 王瑜; 谢芳; 韩文瑜

    2011-01-01

    为获得胸膜肺炎放线杆菌(APP)菌影诱导的仔猪淋巴细胞差异表达基因,本研究应用代表性差异分析技术构建APP菌影免疫前后正、反两个外周血淋巴细胞cDNA差减文库,并对文库中的差异基因进行克隆、测序和生物信息学分析.试验结果表明,正向文库中获得11个表达丰度上调的基因,其中7个基因与已知基因具有相似性,4个为未知新基因,经进一步功能注解发现,正向文库功能基因包括免疫信号传导相关蛋白RhoE、防御相关蛋白糖基转移样酶-1、上皮膜蛋白2、白介素-17和肿瘤免疫相关的周期素依赖性蛋白激酶抑制因子3等,这些功能基因表达丰度升高,可能有助于机体建立抗APP的免疫应答.%To screen differential expression genes in peripheral blood lymphocytes induced by ghost of Actinobacillus pleuropneumoniae, the forward and reverse two subtractive cDNA libraries were constructed from the peripheral blood lymphocytes of piglet vaccinated by bacterial ghost of A. pleuropneumoniae using representational difference analysis technique. The analysis identified differentially expressed transcripts. The results indicated that genes related to immunization signal transduction, disease defence related protein, epithelial membrane protein and interleukin-17, tumor immunity related factors were up-regulated after vaccinated, which may increase the immunity response.

  10. 胸膜肺炎放线杆菌血清8型自转运黏附素基因的克隆测序及功能预测分析%Sequencing and functional analysis of Actinobacillus pleuropneumoniae serotype 8 adhesin gene

    Institute of Scientific and Technical Information of China (English)

    王瑜; 雷连成; 陈创夫; 韩文瑜; 谢芳; 周靓; 邢艳苹; 杨舒心; 何伯萍

    2011-01-01

    为研究胸膜肺炎放线杆菌(APP)三聚体自转运黏附素(TAAs)的功能,以GenBank登录的APP血清5b型自转运黏附素基因5'端的3875bp序列设计引物,通过PCR的方法首次获得APP血清8型运黏附素N端的基因序列片段,测序结果与已知血清型的基因序列和氨基酸推导序列分别进行比对,结果表明与血清7型自转运黏附素N端同源性达到93%,氨基酸推导序列同源性达到97%;与血清5b型自转运黏附素N端同源性达到92%,氨基酸推导序列同源达到100%.经软件分析获得的序列含有与细菌的黏附、聚集和侵入密切相关的Hep_Hag基序,应用马克斯-普朗克研究所的在线分析TAAs的基序和蛋白域的软件daTAA,进行预测并证明所得序列为TAAs,并且具有完整的N段头部序列,有重要功能区具有良好的抗原性.比对的结果为寻找研究APP的定植基序和毒力因素提供了重要基础.%Adhesion is an important pathogenic process for the pathogenesis of bacteria. To study the function of Actinobacillus pleuropneumoniae (APP) autotransporter adhesins (TAAs), the sequence encoding N part of APP serotype 8 TAAs was amplified by PCR with the primers designed based on the APP serotype 5b transshipment adhesion element gene of 3,875 bp sequence (5' end CP000569.1). The sequence was analyzed by software SMART and PFAM which indicated that the sequence contained HepHag base domain, which was closely related with the the bacterial adhesion, aggregation and intrusive, and the TAAs was predicted in the APP serotype 8 sequence by the Max Planck institute of on-line and adhesion grain protein domain software daTAA analysis. The sequence analysis results provided a important basis for further study of the APP colonization and virulence factors.

  11. Evaluation of 5 ' nuclease assay for detection of Actinobacillus pleuropneumoniae

    DEFF Research Database (Denmark)

    Angen, Øystein; Jensen, J.; Lavritsen, D. T.

    2001-01-01

    , nonspecific reactions appeared when testing dilutions of DNA templates or pure cultures of A. pleuropneumoniae, as well as when testing tonsil scrapings from specific-pathogen-free herds. The diagnostic sensitivity, as evaluated with 586 tonsil scrapings from animals infected with A. pleuropneumoniae...

  12. Actinobacillus pleuropneumoniae serovar 8 predominates in England and Wales

    OpenAIRE

    Li, Y; Bossé, JT; Williamson, SM; Maskell, DJ; Tucker, AW; Wren, BW; Rycroft, AN; Langford, PR; BRaDP1T Consortium

    2016-01-01

    This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (BBSRC grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1 and BB/G018553/1) and Zoetis (formerly Pfizer Animal Health) awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology (BRaDP1T) Consortium.

  13. An Unusual Occurrence of Actinobacillosis in Heifers and Cows in A Dairy Herd in Tehran suburb-Iran

    Directory of Open Access Journals (Sweden)

    Atyabi, N.,

    2010-07-01

    Full Text Available An unusual occurrence of actinobacillosis was diagnosed in 4 heifers aged 8-15 months and 2 cattle in a dairy herd with 190 Iranian Holstein breed. Anorexia, dysphagia, drooling of normal or foodtinged saliva and presence of warts-like lesions on the dorsal surface of tongue shaft were observed in a 15-month-old heifer without showing protrusion of tongue or presenting woody tongue and no involvement of either sulcus lingualis or tongue base. In addition to tongue, soft tissues of oral cavity and pharyngeal region including lymph nodes, salivary glands and tonsils were contained multiple whitish nodules. Histopathologically, typical pyogranulomas of actinobacillosis contained radiating eosinophilic clubs surrounded by many neutrophils were diagnosed. Actinobacillus lignieresii was isolated from the lesions in pure culture. Clinical examination of other animals revealed the presence of different degrees of granulomatous abscesses in soft tissues including skin around mandibles in at least 3 heifers aged 8-11 months and 2 cattle. Due to 4 recent droughty years feeding the heifers, dry cows and low milk producing cattle by cheap oat and wheat straw mixed with plant awns could be the cause of this event.

  14. Lesões granulomatosas encontradas em bovinos abatidos para consumo

    Directory of Open Access Journals (Sweden)

    Bianca Tessele

    2014-08-01

    Full Text Available Com o objetivo auxiliar profissionais médico-veterinários no reconhecimento das lesões de bovinos encontradas na linha de inspeção de carnes em matadouros frigoríficos, três condições granulomatosas de bovinos foram pesquisadas e suas semelhanças e diferenças avaliadas. Essas três condições granulomatosas foram actinobacilose (causada por Actinobacillus lignieresii, actinomicose (causada por Actinomyces bovis e mastite estafilocócica (causada por Staphylococcus aureus. Em 505 lesões encontradas em bovinos abatidos para consumo humano, 40 eram uma dessas três lesões granulomatosas: 24 eram actinobacilose, 10 eram actinomicose e seis eram mastite estafilocócica. De um modo geral, os aspectos macro e microscópicos dessas três lesões eram bastante semelhantes, mas suas localizações ajudavam a presumir sua etiologia. A. lignieresii afetou tecidos moles, principalmente língua e linfonodos da cabeça; A. bovis afetou o tecido ósseo, principalmente o da mandíbula; e S. aureus teve a glândula mamária como o tecido alvo. Histologicamente, os granulomas resultantes da infecção por qualquer um desses três agentes continham uma estrutura amorfa, eosinofílica, com clavas irradiadas, localizada centralmente; essa estrutura era rodeada por neutrófilos íntegros e degenerados, que, por sua vez, eram cercados por um manto de macrófagos epitelioides e ocasionais células gigantes multinucleadas. Esses mantos de macrófagos eram irregularmente infiltrados por linfócitos e plasmócitos que tendiam a se acumular na periferia da lesão, que era cercada por uma cápsula de tecido conjuntivo. Dependendo da aplicação do método de coloração adequado, o agente etiológico podia ser visto em cada um dos três tipos de lesão granulomatosa. No caso da mastite estafilocócica, cocos intralesionais foram observados tanto nas colorações por HE como nas de Gram, nessa última como cocos gram-positivos. O agente da actinobacilose

  15. A multiplexed immunoassay for detection of antibodies to Actinobacillus pleuropneumoniae (App) in pigs

    DEFF Research Database (Denmark)

    Berger, Sanne Schou; Boas, Ulrik; Andresen, Lars Ole

    2014-01-01

    our diagnostic tools, we are currently developing a novel indirect fluorescent microsphere immunoassay that can facilitate simultaneous detection of antibodies towards multiple App serovars within a single serum sample volume. The multiplex immunoassay is based on Luminex technology (8) and has...

  16. Comparison of high and low virulence serotypes of Actinobacillus pleuropneumoniae by quantitative real-time PCR

    DEFF Research Database (Denmark)

    Schou, Kirstine Klitgaard; Angen, Øystein; Boye, Mette

    of high virulence while serotype 6 strains are normally found to be less pathogenic. To gain an understanding of the differential virulence of serotype 2 and 6, the expression of a panel of Ap genes during infection of porcine epithelial lung cells (SJPL) were examined by quantitative real-time PCR (qPCR...... to be important for early establishment of the bacteria in the host were examined by qPCR. The genes examined were apfA, coding for a subunit of Type IV pili, kdsB coding for a gene involved in lippopolysacceride biosynthesis, and pgaB which is involved in biofilm formation, all three believed to be important...... with respect to host cells adhesion. Also included in the analysis were the capsular gene, cpxB, the RTX toxin genes apxII, and apxIV and the gene exbD, involved in binding of iron from host cells. Finally, three previously validated reference genes, glyA, pykA and tpiA were included for normalization of the qPCR...

  17. Concurrent host-pathogen gene expression in the lungs of pigs challenged with Actinobacillus pleuropneumoniae

    DEFF Research Database (Denmark)

    Brogaard, Louise; Schou, Kirstine Klitgaard; Heegaard, Peter M. H.;

    2015-01-01

    4, CD14, MD2, LBP, MYD88) in response to A. pleuropneumoniae. Significant up-regulation of proinflammatory cytokines such as IL1B, IL6, and IL8 was observed, correlating with protein levels, infection status and histopathological findings. Host genes encoding proteins involved in iron metabolism...

  18. Putative biomarkers for evaluating antibiotic treatment: an experimental model of porcine Actinobacillus pleuropneumoniae infection

    DEFF Research Database (Denmark)

    Lauritzen, B.; Lykkesfeldt, J.; Skaanild, M.T.;

    2003-01-01

    the animals received a single dose of either danofloxacin (2.5 mg/kg) or tiamulin (10 mg/kg). To test the discriminative properties of the biomarkers, the dosage regimens were designed with an expected difference in therapeutic efficacy in favour of danofloxacin. Accordingly, the danofloxacin-treated pigs...... recovered clinically within 24h after treatment, whereas tiamulin-treated animals remained clinically ill until the end of the study, 48 h after treatment. A similar Picture was seen for the biomarkers of infection. During the infection period, plasma C-reactive protein (CRP), interleukin-6 and haptoglobin...... increased, whereas plasma zinc, ascorbic acid and alpha-tocopherol decreased. In the danoffoxacin-treated animals, CRP, interleukin-6, zinc, ascorbic acid and alpha-tocopherol reverted significantly towards normalisation within 24h of treatment. In contrast, signs of normalisation were absent (CRP, zinc...

  19. Differences in iron acquisition from human haemoglobin among strains of Actinobacillus actinomycetemcomitans

    DEFF Research Database (Denmark)

    Hayashida, H.; Poulsen, Knud; Kilian, Mogens

    2002-01-01

    . actinomycetemcomitans strains examined harboured a single genomic sequence with homology to the hgpA gene encoding haemoglobin-binding protein A in Haemophilus influenzae. However, in all three strains belonging to the JP2 clone and in one serotype e strain hgpA was a pseudogene. Seven other strains possessed...

  20. Actinobacillus pleruropneumoniae transcriptome analysis during early infection - coping with a hostile environment

    DEFF Research Database (Denmark)

    Schou, Kirstine Klitgaard; Rundsten, Carsten Friis; Jensen, Tim Kåre

    2011-01-01

    . Methods: The local in vivo genetic response of Ap during the early phase of infection in porcine lungs was detailed using pangenomic microarray analysis. The global transcriptional patterns of Ap serotype 2 and 6 isolated from lung tissue biopsies of 25 experimentally infected pigs were compared at four...

  1. Host Cell Contact-Induced Transcription of the Type IV Fimbria Gene Cluster of Actinobacillus pleuropneumoniae

    NARCIS (Netherlands)

    Boekema, B.K.H.L.; Putten, J.P.M.; Stockhofe-Zurwieden, N.; Smith, H.E.

    2004-01-01

    Type IV pili (Tfp) of gram-negative species share many characteristics, including a common architecture and conserved biogenesis pathway. Much less is known about the regulation of Tfp expression in response to changing environmental conditions. We investigated the diversity of Tfp regulatory system

  2. Comparison of virulence of different Actinobacillus pleuropneumoniae serotypes and biotypes using an aerosol infection model

    DEFF Research Database (Denmark)

    Jacobsen, Mariann Juul; Nielsen, Jens Peter; Nielsen, Ragnhild

    1996-01-01

    . The pigs were sacrificed 24 h after aerosol exposure and lung lesions were evaluated. In pigs exposed to aerosols of suspensions containing 10(4) CFU/ml of serotypes 2, 5b and 6, a number of 5-10 lesions of haemorrhagic necrotizing pneumonia were induced, For the biotype 2 strain the dose creating similar...... lesions was 10(9) CFU/ml. Repeated experiments confirmed these results showing similar virulence of serotypes 2, 5b and 6 whereas the biotype 2 strain proved less virulent, The aerosol infection model allowed a comparison of the number of A. pleuropneumoniae CFU/liter air which were necessary to induce...

  3. An atypical biotype I Actinobacillus pleuropneumoniae serotype 13 is present in North America

    DEFF Research Database (Denmark)

    Perry, Malcolm B.; Angen, Øystein; MacLean, Leann L.

    2012-01-01

    analysis of the capsular polysaccharide (CPS) and lipopolysaccharide (LPS) of a representative strain revealed that the CPS is almost identical to that of the reference strain of serotype 13, having a slightly higher degree of glycose O-acetylation. However, it produces an O-PS within the LPS antigenically...... and structurally identical with that of the reference strain of A. pleuropneumoniae serotype 10. The O-PS was characterized as a homopolymer of 1,2 linked β-d-galactofuranosyl residues, a structure unrelated to that of the O-PS produced by the reference strain of serotype 13. Strains from Canada and United States...

  4. Microevolution and Patterns of Dissemination of the JP2 Clone of Aggregatibacter (Actinobacillus) actinomycetemcomitans

    DEFF Research Database (Denmark)

    Haubek, Dorte; Poulsen, Knud; Kilian, Mogens

    2007-01-01

    belonging to the JP2 clone had a number of point mutations, particularly in the pseudogenes hbpA and tbpA. Characteristic mutations allowed isolates from individuals from the Mediterranean area and from West Africa, including the Cape Verde Islands, to be distinguished. The patterns of mutations indicate...

  5. Evaluation of a multiplex PCR test for simultaneous identification and serotyping of Actinobacillus pleuropneumoniae serotypes 2, 5, and 6

    DEFF Research Database (Denmark)

    Jessing, Stine Graakjær; Angen, Øystein; Inzana, Tomas J.

    2003-01-01

    , and 6 were combined with the already existing species-specific primers used in a PCR test based on the omlA gene. The PCR test was evaluated with serotype reference strains of A. pleuropneumoniae as well as 182 Danish field isolates previously serotyped by latex agglutination or immunodiffusion. For all...... that cross-reacted by the latex agglutination test were of serotype 2, 5, or 6. Determination of the serotype by PCR represents a convenient and specific method for the serotyping of A. pleuropneumoniae in diagnostic laboratories....

  6. Transcriptional Portrait of Actinobacillus pleuropneumoniae during Acute Disease - Potential Strategies for Survival and Persistence in the Host

    DEFF Research Database (Denmark)

    Schou, Kirstine Klitgaard; Rundsten, Carsten Friis; Jensen, Tim Kåre

    2012-01-01

    was monitored during the acute phase of infection in its natural host. Methodology/Principal Findings Bacterial expression profiles of A. pleuropneumoniae isolated from lung lesions of 25 infected pigs were compared in samples taken 6, 12, 24 and 48 hours post experimental challenge. Within 6 hours, focal......, fibrino hemorrhagic lesions could be observed in the pig lungs, indicating that A. pleuropneumoniae had managed to establish itself successfully in the host. We identified 237 differentially regulated genes likely to encode functions required by the bacteria for colonization and survival in the host....... This group was dominated by genes involved in various aspects of energy metabolism, especially anaerobic respiration and carbohydrate metabolism. Remodeling of the bacterial envelope and modifications of posttranslational processing of proteins also appeared to be of importance during early infection...

  7. Transmission of Actinobacillus pleuropneumoniae in pigs under field-like conditions: emphasis on tonsillar colonisation and passively acquired colostral antibodies

    DEFF Research Database (Denmark)

    Vigre, Håkan; Angen, Øystein; Barfod, K.;

    2002-01-01

    consisted of the offspring from five sows originating from a conventional pig herd. The sows were transferred to isolated research facilities before farrowing. A. pleuropneumoniae was detected on the tonsils of all sows. After a nursing period of 3 weeks, the pigs were weaned and reared isolated from other...... the proportion of pigs with detectable levels of colostral antibodies to the different serotypes of A. pleuropneumoniae was declining. Since these two events take place in the same age period, we expect a possible biological association between the level of the passive immunity and the degree of tonsillar...

  8. Effect of tulathromycin on the carrier status of Actinobacillus pleuropneumoniae serotype 2 in the tonsils of pigs

    DEFF Research Database (Denmark)

    Angen, Øystein; Andreasen, M.; Nielsen, E.O.

    2008-01-01

    PCR test on tonsil scrapings and they were divided into three groups. The pigs in group I were treated subcutaneously with 2.5 mg/kg tulathromycin on day 0, the pigs in group 2 were treated with 2.5 mg/kg tulathromycin on days 0 and 4, and the pigs in group 3 were left untreated as controls. The pigs...... were tested by PCR on tonsil scrapings on days 0, 4, 11 and 33, and on day 33 all the animals were euthanased. There were no significant differences between the numbers of PCR-positive animals in the three groups on any of the sampling dates....

  9. DEVELOPMENT AND EVALUATION OF A SELECTIVE AND INDICATIVE MEDIUM FOR ISOLATION OF ACTINOBACILLUS-PLEUROPNEUMONIAE FROM TONSILS

    DEFF Research Database (Denmark)

    Jakobsen, Marianne; Nielsen, Jens

    1995-01-01

    In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A. pleuropn......In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A...

  10. Comparative profiling of the transcriptional response to iron restriction in six serotypes of Actinobacillus pleuropneumoniae with different virulence potential

    DEFF Research Database (Denmark)

    Schou, Kirstine Klitgaard; Friis, Carsten; Angen, Øystein

    2011-01-01

    of virulence genes. We used a pan-genomic microarray to study the transcriptional response to iron restriction in vitro in six serotypes of A. pleuropneumoniae (1, 2, 3, 5b, 6, and 7), representing at least two levels of virulence. Results In total, 45 genes were significantly (p

  11. The pharmacodynamic effect of amoxycillin and danofloxacin against Actinobacillus pleuropneumoniae in an in-vitro pharmacodynamic model

    DEFF Research Database (Denmark)

    Lindecrona, R.H.; Friis, C.; Jensen, N.E.

    1999-01-01

    the experiments, which is consistent with time > Mle as the most important parameter of pharmacodynamic effect of beta-lactam drugs. For danofloxacin maximal bactericidal effect initially was observed at peak concentrations of at least eight times the we. The pharmacodynamic effect was dependent on the peak...

  12. The genetic organisation of the capsule biosynthesis region of Actinobacillus pleuropneumoniae serotypes 1, 6, 7, and 12

    DEFF Research Database (Denmark)

    Jessing, Stine Graakjær; Ahrens, Peter; Inzana, Thomas J.

    2008-01-01

    C of A.pleuropneumoniae serotypes 2, 6, 7, and 8 contained a high degree of homology. At the amino acid level Cps6D revealed a high degree of homology to Cps8D, whereas Cps7D contained a high degree of homology to the Cps2D. The deduced gene product of the partially sequenced cps6E gene showed...

  13. Activity of antibodies against Salmonella dublin, Toxoplasma gondii, or Actinobacillus pleuropneumoniae in sera after treatment with electron beam irradiation or binary ethylenimine

    DEFF Research Database (Denmark)

    Kyvsgaard, N.C.; Lind, Peter; Preuss, T.;

    1996-01-01

    was used as an estimate for the relative posttreatment activity. For a Toxoplasma gondii indirect enzyme-linked immunosorbent assay (ELISA) and agglutination assay as well as for a Salmonella dublin indirect ELISA, the posttreatment activity was more than 89% of the pretreatment activity when the samples...... inactivation, especially when used in indirect ELISA or in the T. gondii agglutination assay....

  14. Intra-unit correlations in seroconversion to Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae at different levels in Danish multi-site pig production facilities

    DEFF Research Database (Denmark)

    Vigre, Håkan; Dohoo, I.R.; Stryhn, H.;

    2004-01-01

    2) and Mycoplasma hyopneumoniae (Mh). Based on the estimated variances, three newly described computational methods (model linearisation, simulation and linear modelling) and the standard method (latent-variable approach) were used to estimate the correlations (intra-class correlation components......, ICCs) between pigs in the same production unit regarding seroconversion. Substantially different values of ICCs were obtained from the four methods. However, ICCs obtained by the simulation and the model linearisation were quite consistent. Data used for estimation were collected from 1161 pigs from...

  15. Construction of genome library of Actinobacillus pleuropneumoniae%猪胸膜肺炎放线杆菌基因组文库的构建

    Institute of Scientific and Technical Information of China (English)

    杨建德; 刘燕霏; 徐军

    2008-01-01

    由猪胸膜肺炎放线杆菌(APP)引起的猪传染性胸膜肺炎是猪最重要的呼吸道传染病之一.为研究该细菌表面蛋白的结构与功能,选取APP代表菌株,提取其基因组DNA,用TSP5091随机消化成3~8 kb的片段并回收,连接到预先消化的λZAPII载体上,经过包装、扩增和滴定后,成功地构建了APP基因组文库.

  16. Prevalencia de periodontite juvenil localizada, generalizada e incipiente e presença de Actinobacillus actinomycetemcomitans em individuos de 15 a 25 anos de idade

    OpenAIRE

    Jose Roberto Cortelli

    2000-01-01

    As periodontites que atingem indivíduos jovens podem ser classificadas em periodontite pré-pubertal, periodontite juvenil localizada ou generalizada e periodontite incipiente. O estudo da prevalência destas patologias apresenta extensa variação nos valores encontrados por diversos autores. O objetivo do presente estudo foi avaliar a condição clínica periodontal de indivíduos entre 15 e 25 anos de idade da região do Vale do Paraíba, estado de São Paulo, nos quais observou-se a prevalência de...

  17. An Actinobacillus pleuropneumoniae PCR typing system based on the apx and omlA genes - evaluation of isolates from lungs and tonsils of pigs

    DEFF Research Database (Denmark)

    Gram, T.; Ahrens, Peter; Andreasen, Morten;

    2000-01-01

    . The PCR typing system was tested on 102 field strains of A. pleuropneumoniae isolated from lungs of diseased pigs. The serotyping results of the investigated field strains were in agreement with the apr and omlA gene patterns found in the reference strains of the bacteria, with the exception of the oml......A gene of five strains of serotype 8. To examine the apx and omlA gene pattern of tonsil isolates, the PCR typing system was tested on a total of 280 A. pleuropneumoniae field strains isolated from tonsils of pigs. Agreement between serotyping and DNA typing was found in 96% of the isolates using the apx...... gene patterns and in 89% of the isolates using the omlA gene. The same serotype specific apx/omlA gene pattern was thus found in the majority of the tonsil isolates and in isolates from diseased lungs. Most of the differences in the omlA gene were found in 18 tonsil isolates of serotype 12. The oml...

  18. Identification of αLβ2, αMβ2, and αXβ2 integrins as receptors for Actinobacillus actinomycetemcomitans leukotoxin

    DEFF Research Database (Denmark)

    Reinholdt, Jesper; Poulsen, Knud; Kilian, Mogens;

    associated with a highly aggressive form of disease in adolescents of African descent (1). An earlier report (2) identified the β2 integrin LFA-1 (αLβ2) as a cell surface receptor for LtxA. Whether the LtxA-reactive site involves αL (CD11a), β2 (CD18), or both of these subunits, is unknown.  Notably...

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-0294 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0294 ref|ZP_04753087.1| hypothetical protein AM305_07508 [Actinobacillus... minor NM305] gb|EER47509.1| hypothetical protein AM305_07508 [Actinobacillus minor NM305] ZP_04753087.1 0.038 31% ...

  20. Inducción experimental de epididimitis en ovinos por inoculación intrauretral con Actinobacillus seminis: estudio bacteriológico, serológico e histopatológico

    OpenAIRE

    Jorge Acosta Dibarrat; Efrén Díaz Aparicio; Beatriz Arellano Reynoso; Víctor Rubén Tenorio Gutiérrez; Jorge Tórtora Pérez

    2006-01-01

    Con el objetivo de inducir la infección experimental de A. seminis, se utilizaron 18 corderos de seis meses de edad: 4 testigos negativos, otros 3 se inocularon con A. seminis vía intraepididimal (IE) y 11 fueron inoculados por vía intrauretral (IU) como sigue: cuatro recibieron factor liberador de gonadotropinas (GnRH) cinco días previos al desafío, cuatro recibieron etilenglicol por vía IE 24 h previas al desafío y tres no recibieron tratamiento previo. De los cuatro testigos negativos, dos...

  1. 21 CFR 522.313a - Ceftiofur crystalline free acid.

    Science.gov (United States)

    2010-04-01

    ... Actinobacillus pleuropneumoniae, Pasteurella multocida, Haemophilus parasuis, and Streptococcus suis. (iii... fever, pneumonia) associated with Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni.... (2) Cattle. The formulation described in paragraph (a)(2) of this section is used as follows:...

  2. 猪胸膜肺炎放线杆菌血清1型信号标签诱导突变体库的构建%CONSTRUCTION AND EVALUATION OF SIGNATURE-TAGGED MUTAGENESIS LIBRARIES OF ACTINOBACILLUS PLEUROP NEUMONIAE SEROTYPE

    Institute of Scientific and Technical Information of China (English)

    赫明雷; 刘慧芳; 刘思国; 司薇; 王春来; 杨金国; 杜艳芬; 王聃

    2009-01-01

    本研究以自杀性质粒pUT携带含有信号标签的Mini-Tn5转座子对猪胸膜肺炎放线杆菌血清1型菌进行转座诱变,构建了带有12对特异性信号标签的Mini-Tn5转座子的pUT自杀质粒,转化到供体菌E.coliβ2155后,利用双亲本滤膜杂交法与受体猪胸膜肺炎放线杆菌血清1型菌(APP1)进行接合转移,构建并优化了接合转移体系.利用抗性和营养缺陷培养平板筛选得到接合突变体,通过抗性通用引物与12个特异标签引物和胸膜肺炎放线杆菌毒素IV(ApxIV)鉴定引物分别对这些突变体进行了PCR鉴定和测序验证.结果表明,经过加入标签的MinI-Tn5转座子可以通过接合转移的方式从供体菌E.COliβ2155中插入到APP1基因组当中,并成功构建了12个含有特异信号标签的重组质粒,获得了APP1的12个转座突变体库,经筛选鉴定后得到561个突变株.这为研究APP1的功能基因和筛选特定突变株提供了必要的基础.

  3. Cultivable bacterial diversity of terrestrial thermal spring of Unkeshwar, India

    Directory of Open Access Journals (Sweden)

    Anupama Prabhakarrao Pathak

    2014-12-01

    Full Text Available Ten different thermotolerent bacteria were isolated from terrestrial thermal spring of Unkeshwar in Nanded district of Maharashtra (India. These isolates were characterized by morphological characters, microscopic features, biochemical pattern and  physiological attributes. These isolates were identified as Bacillus licheniformis (APP7, Bacillus megaterium (APP8, Actinobacillus hominis (APP9, Lysinibacillus sphaericus (APP10, Paenibacillus alvei (APP11, Bacillus simplex (APP12, Actinobacillus seminis (APP13, Pseudomonas fragii (APP14, Staphylococcus cohnii (APP15 and Streptococcus thermophilus (APP16. These isolates belonged to class Firmicutes and Gamma proteobacteria and showed production of biotechnologically important thermostable hydrolytic enzymes such as caseinase, amylase, gelatinase, urease and lipase.

  4. AcEST: BP919111 [AcEST

    Lifescience Database Archive (English)

    Full Text Available ments: (bits) Value sp|Q1L673|VDRB_DANRE Vitamin D3 receptor B OS=Danio rerio GN=vdr... 32 1.5 sp|A6VMD5...E+ Sbjct: 391 EH 392 >sp|A6VMD5|PRIB_ACTSZ Primosomal replication protein n OS=Actinobacillus succinogenes (

  5. Detection of the Host Immune Response to Burkholderia mallei Heat-Shock Proteins GroEL and DnaK in a Glanders Patient and Infected Mice

    Science.gov (United States)

    2007-01-01

    infected animal are the most common methods of acquiring glanders because the organism can be trans- mitted through droplets or saliva . In horses...host cell. A GroEL-like protein in Actinobacillus actinomycetemcomitans, a patho- gen associated with periodontal disease, was found in extracellular

  6. Bacterial endocarditis due to eikenella corrodens: A case report

    Directory of Open Access Journals (Sweden)

    Mahapatra A

    2003-01-01

    Full Text Available Of all the causes of bacterial endocarditis, HACEK group consisting of Haemophilus, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella Kingae are rare causative agents. We report a case of bacterial endocarditis by E. corrodens, which is one of the members of the HACEK group.

  7. Dicty_cDB: VHG877 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available lk Homology vs CSM-cDNA Score E Sequences producing significant alignments: (bits) Value... E Sequences producing significant alignments: (bits) Value N ( BJ440834 ) Dictyo... Sequences producing significant alignments: (bits) Value CP000687_912( CP000687 |pid:none) Actinobacillus p

  8. Arabidopsis CDS blastp result: AK243490 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243490 J100073N23 At2g20690.1 68415.m02429 lumazine-binding family protein SP|P50...854 Riboflavin synthase alpha chain (EC 2.5.1.9) {Actinobacillus pleuropneumoniae}; contains Pfam profile PF00677: Lumazine binding domain 2e-56 ...

  9. Measurement of bacterial gene expression in vivo by laser capture microdissection and quantitative real-time RT-PCR

    DEFF Research Database (Denmark)

    Schou, Kirstine Klitgaard; Jensen, Tim Kåre; Angen, Øystein

    2007-01-01

    Due to the relative small number of bacterial pathogens present in an infected host, exploration of pathogen gene expression in vivo is challenging. This study reports the development of a protocol for quantifying bacterial gene expression in vivo in Actinobacillus pleuropneumoniae using laser ca...

  10. Identifikasi Bakteri Dan Tes Sensitifitas Terhadap Ciprofloxacin Pada Periodontitis Kronis

    OpenAIRE

    Irene Edith Riuwpassa, Dr.drg. M.Si

    2011-01-01

    Berdasarkan hasil penelitian diperoleh distribusi kuman sebagai berikut: Porhyromonas gingivalis(52), Streptococcus(47), Actinobacillus actinomycetemcomitans(31), Pseudomonas(9), Staphylococcus(11), Klebsiela pneumonia(5), Escherichia colli(2), Alkaigines fascialis(2) dan Enternobacter aerogenus(2) Terdapat P.AERUGINOSA YANG RESISTEN TERHADAP CIPROFLOXACIN SEBESAR 50% (4) DARI 8 SAMPEL. Periodontitis adalah tipe umum penyakit periodontal yang disebabkan oleh perluasan radang tahap awal pad...

  11. The concentration of apolipoprotein A-I decreases during experimentally induced acute-phase processes in pigs

    DEFF Research Database (Denmark)

    Carpintero, R.; Pineiro, M.; Andres, M.

    2005-01-01

    In this work, apolipoprotein A-I (ApoA-I) was purified from pig sera. The responses of this protein after sterile inflammation and in animals infected with Actinobacillus pleuropneumoniae or Streptococcus suis were investigated. Decreases in the concentrations of ApoA-I, two to five times lower...

  12. The porcine systemic response to pleuropneumonia studied by transcriptional profiling of liver and tracheobronchial lung lymph nodes using multiplexed mRNA-Seq

    DEFF Research Database (Denmark)

    Hedegaard, Jakob; Schou, Kirstine Klitgaard; Skovgaard, Kerstin

    2010-01-01

    Actinobacillus pleuropneumoniae (Ap) is a gram-negative bacterium that causes porcine pleuropneumonia, which is a widespread, highly contagious and often fatal respiratory disease in swine. A total of 44 pigs were experimentally inoculated with Ap serotype 2 or 6 and samples of liver and tracheob......Actinobacillus pleuropneumoniae (Ap) is a gram-negative bacterium that causes porcine pleuropneumonia, which is a widespread, highly contagious and often fatal respiratory disease in swine. A total of 44 pigs were experimentally inoculated with Ap serotype 2 or 6 and samples of liver...... and tracheobronchial lung lymph nodes were collected 6, 12, 24 and 48 hours after experimental inoculation, as well as from six non-inoculated control pigs. Transcriptional profiles of the liver samples have been generated by preparation of 12-plexed mRNA-Seq libraries followed by sequencing on an Illumina GAIIx (51...

  13. Ischemic Stroke and Septic Shock After Subacute Endocarditis Caused by Haemophilus parainfluenzae: Case Report

    OpenAIRE

    Menegueti, Mayra Goncalves; Machado-Viana, Jaciara; Gaspar, Gilberto Gambero; Nicolini, Edson Antonio; Basile-Filho, Anibal; Auxiliadora-Martins, Maria

    2016-01-01

    Haemophilus parainfluenzae, which belongs to the HACEK (Haemophilus ssp, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella kingae) group, is a rare cause of subacute endocarditis and may lead to ischemic stroke. A 65-year-old female patient previously diagnosed with rheumatic valve disease was submitted to surgical mitral valve repair in 1996. Physical examination did not reveal any murmurs; physical examination of the lungs and abdomen was norma...

  14. Cardiobacterium hominis endocarditis: A case report and review of the literature

    OpenAIRE

    Andrew Walkty

    2005-01-01

    The present case report describes the clinical course of a patient who presented with Cardiobacterium hominis endocarditis. A review of the literature follows the case presentation. C hominis, a fastidious Gram-negative bacillus, is a member of the HACEK group of microorganisms (Haemophilus species, Actinobacillus actinomycetemcomitans, C hominis, Eikenella corrodens and Kingella kingae). Endocarditis caused by C hominis is uncommon and generally follows a subacute course. Patients may presen...

  15. Cardiobacterium hominis-induced acute dacryocystitis and lacrimal abscess

    Directory of Open Access Journals (Sweden)

    Guru Prasad Manderwad

    2014-01-01

    Full Text Available Cardiobacterium hominis is a member of the HACEK (Haemophilus sp., Actinobacillus actinomycetemcomitans, C. hominis, Eikenella corrodens, and Kingella kingae group commonly associated with endocarditits and is normally present in the respiratory tract. We describe the first case of acute dacryocystitis with lacrimal abscess caused by C. hominis along with a brief review of the literature. The patient responded to oral and topical ciprofloxacin after incision and drainage and awaits dacryocystorhinostomy.

  16. Prevalence of Periodontal Pathogens in Dental Plaque of Children

    OpenAIRE

    Gafan, Gavin P.; Lucas, Victoria S.; Roberts, Graham J; Petrie, Aviva; Wilson, Michael; Spratt, David A.

    2004-01-01

    Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Tannerella forsythensis have been implicated as the main etiological agents of periodontal disease. The purpose of this work was to estimate the prevalence of these organisms in plaque from children without gingivitis (group 1; n = 65) and from those with gingivitis (group 2; n = 53). Extracted DNA from plaque was subjected to two rounds of PCR targeting the 16S rRNA gene using both universal primers and species-specific prim...

  17. Antibacterian in vitro determinatíon of Menthostachys mollis (Muña) opposite to oral bacterial stomatological importance

    OpenAIRE

    Díaz L., Karin; Bachiller en Odontología. Facultad de Odontología, Universidad Nacional Mayor de San Marcos, Lima, Perú.; Moromi N., Hilda; Departamento Académico de Ciencias Básicas,Microbiología. Facultad de Odontología, Universidad Nacional Mayor de San Marcos, Lima, Perú.

    2014-01-01

    With the aim to determine the antimícrobial action of the essential oil of Methosthachys mollis (Muña); in design at random one strains standard ATCC of Streptococcus mutans, Lactobacillus sp, Fusobacterium nucleatum, Actinobacillus actinomicetencomitans and Actinomyces sp, to: Amoxicilina (positive witness), essential oil of Methosthachys mollis and, distilled water (negative witness); to measure the halos of antitmicrobial action. For the mentioned bacteria one found, respectively, in Amoxi...

  18. Influencia del estrógeno en la enfermedad periodontal: revisión de literatura

    OpenAIRE

    2015-01-01

    La etiología de la enfermedad periodontal está bien definida, dentro de los agentes etiológicos que la causan podemos citar algunos microorganismos subgingivales como: Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, Actinobacillus actinomycetemcomitans y espiroquetas. La susceptibilidad del huésped a estos agentes bacterianos también tiene un papel importante dentro del progreso y prevalencia de la enfermedad periodontal. Dentro de los factores de riesgo asociados con ...

  19. Aspectos clínicos, radiográficos e microbianos de uma família com expressiva prevalência de doença periodontal = Clinical, radiographics and microbiological aspects of a family with expressive prevalence of periodontal disease

    Directory of Open Access Journals (Sweden)

    Querido, Silvia Maria Rodrigues

    2006-01-01

    Full Text Available A periodontite agressiva é caracterizada por uma rápida perda de inserção conjuntiva e destruição óssea, podendo ocorrer na forma localizada ou generalizada. Na periodontite agressiva localizada ocorre o comprometimento, principalmente, dos dentes incisivos e primeiros molares permanentes. É a forma de doença periodontal mais associada com Actinobacillus actinomycetemcomitans. De modo distinto, a forma generalizada da periodontite agressiva afeta vários dentes em um ou ambos os arcos dentários. O objetivo do presente estudo foi estabelecer através de exames clínicos, radiográficos e microbianos o diagnóstico de uma família com expressiva prevalência de doença periodontal. Foram incluídos 5 indivíduos da mesma família, os quais foram submetidos à exame clínico periodontal e exame radiográfico de todos os dentes. Para a análise microbiológica foram selecionados no mínimo dois dentes para cada indivíduo, e a presença de Actinobacillus actinomycetemcomitans foi determinada pelo método de cultura bacteriana e reação em cadeia da polimerase. Dos indivíduos examinados, dois receberam o diagnóstico de periodontite incipiente, dois de periodontite agressiva localizada e um de periodontite agressiva generalizada. O Actinobacillus actinomycetemcomitans de máxima leucotoxicidade foi detectado em todos os indivíduos. Com base nestes resultados observou- se que os indivíduos apresentavam a mesma característica microbiana e diferenças na expressão e severidade da doença periodontal, sugerindo que outros fatores poderiam estar presentes modificando a manifestação clínica da doença

  20. Eikenella corrodens: Patogénesis y aspectos clínicos

    OpenAIRE

    Rubén Darío Jaramillo; Paola Suárez; Beatriz Barraza; Paulina Lara; Luis Teherán; José Edgardo Escamilla

    2006-01-01

    El ambiente microbiológico oral es único y tiene una dinámica compleja. Se calcula que cerca de 500 especies de bacterias habitan la cavidad oral humana, y alrededor de 22 géneros son los predominantes. Las bacterias que se aíslan con más frecuencia de los sitios infectados de la cavidad oral, y que son también patógenos potenciales, forma un grupo pequeño de microorganismos gramnegativos, entre los que se incluyen los siguientes: Actinobacillus actinomycetemcomitans, Bacteroides forsythus, C...

  1. HELICOBACTER PYLORI EN LA FLORA BACTERIANA ORAL

    OpenAIRE

    Moromi Nakata, Hilda; Departamento Académico Ciencias Básicas Estomatológicas. Facultad de Odontología. UNMSM.

    2014-01-01

    No hay duda de la relación existente entre enfermedades orales con otras enfermedades sistémicas. En tal contexto, las bacterias de la flora bacteriana oral, que alcanzan alrededor de 350 especies, para la mayoría de tales bacterias no se ha demostrado un rol específico, conociéndose sí una clara relación entre los Estreptococos orales (Streptococcus sanguis, Strecoccus mutans, Streptococcus sobri nus) y Actinobacillus actinomycetencomitans, entre otros, con la endocarditis bacteriana; así co...

  2. Investigations of representation of certain bacteria strains in lungs of pigs with pneumonia

    Directory of Open Access Journals (Sweden)

    Žutić Milenko

    2009-01-01

    Full Text Available The objective of the investigations described in this paper was to carry out the identification and to establish the incidence of certain strains of bacteria that take part in the etiopathogenesis of pig infections. The investigations covered a total of 237 pathoanatomically altered lungs of expired pigs. Sampling was done during visits to pig farms, most often in situations when it had been necessary to resolve occurring respiratory infections. The samples were examined in a laboratory for the presence of bacterial causes using standard and commercial methods of microbiological diagnostics. For this purpose, the samples were sown on corresponding nutritive bases (blood agar, MacConkey agar, nutritive agar, BHI agar, Baird Parker agar. For the primary isolation of Actinobacillus pleuropneumoniae and Haemophilus parasuis, agar with 5-10% sheep's blood was used and the culture of the strain Staphylococcus aureus which serves as a source of V factor, and for subcultivation of these causes, chocolate agar was used with PolyVitex. In the isolated bacteria, following investigations of morphological and culture characteristics, biochemical identification was performed using the commercial tests BBL Crystal GP ID Kit, E/N ID Kit, Api 20 Strep and Slidex Staph Plus. From the total of 237 examined lung samples, 13 bacteria strains were isolated from 193 samples (81.43%. Among breeding pigs, 112 lung samples were examined and the presence of bacteria was established in 92 (82.14%, while the presence of bacteria was established in 101 samples (80.8% of 125 examined lung samples from fattening pigs. Two bacteria strains were dominant among the spectrum of lung microorganisms: Pasteurella multocida (32.64% and Actinobacillus pleuropneumoniae (29.02%, or, these two bacteria strains in total accounted for 61.66% of all strains isolated in pure culture. The participation of the other 11 bacteria strains ranged from 0.52-9.84%. Observed according to production

  3. Antimicrobial profile screening of two oils of Copaifera genus

    OpenAIRE

    F.A. Pieri; V.O. Silva; SOUZA, C. de F.; Costa,J.C.M.; L.F. Santos; Moreira,M.A.S.

    2012-01-01

    O objetivo deste estudo foi identificar a atividade inibitória de óleos de copaíba sobre o crescimento dos micro-organismos: Shigella flexneri, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Citrobacter freundi, Staphylococcus aureus, Actinobacillus pleuropneumoniae e Haemophilus parasuis. Foi realizado um teste de difusão em ágar com duas soluções a 10% de óleos de copaíba obtidos de duas diferentes espécies de copaíba (Copaifera officinalis e C. langsdor...

  4. Identification of Dominant Immunogenic Bacteria and Bacterial Proteins in Periodontitis

    DEFF Research Database (Denmark)

    Agerbæk, Mette Rylev; Haubek, Dorte; Birkelund, Svend

    Marginal periodontitis is considered an infectious disease that triggers host inflammatory responses resulting in destruction of the periodontium. A complex biofilm of bacteria is associated with periodontitis. Some species have been identified as putative pathogens such as Porphyromonas gingivalis...... (P.g) and Actinobacillus actinomycetemcomitans (A.a), but the identity of dominate immunogens of these bacteria is poorly elucidated. The aim of the study was to identify dominant immunogenic proteins of P.g and A.a in patients suffering from chronic and aggressive periodontitis by proteomic analysis...... will be able to identify immunodominant proteins and potentially important virulence factors of putative periodontal pathogens....

  5. Impact of CDT Toxin on Human Diseases

    Directory of Open Access Journals (Sweden)

    Tiphanie Faïs

    2016-07-01

    Full Text Available Cytolethal distending toxin (CDT is found in Gram-negative bacteria, especially in certain Proteobacteria such as the Pasteurellaceae family, including Haemophilus ducreyi and Aggregatibacter (Actinobacillus actinomycetemcomitans, in the Enterobacteriaceae family and the Campylobacterales order, including the Campylobacter and Helicobacter species. In vitro and in vivo studies have clearly shown that this toxin has a strong effect on cellular physiology (inflammation, immune response modulation, tissue damage. Some works even suggest a potential involvement of CDT in cancers. In this review, we will discuss these different aspects.

  6. Impact of CDT Toxin on Human Diseases

    Science.gov (United States)

    Faïs, Tiphanie; Delmas, Julien; Serres, Arnaud; Bonnet, Richard; Dalmasso, Guillaume

    2016-01-01

    Cytolethal distending toxin (CDT) is found in Gram-negative bacteria, especially in certain Proteobacteria such as the Pasteurellaceae family, including Haemophilus ducreyi and Aggregatibacter (Actinobacillus) actinomycetemcomitans, in the Enterobacteriaceae family and the Campylobacterales order, including the Campylobacter and Helicobacter species. In vitro and in vivo studies have clearly shown that this toxin has a strong effect on cellular physiology (inflammation, immune response modulation, tissue damage). Some works even suggest a potential involvement of CDT in cancers. In this review, we will discuss these different aspects. PMID:27429000

  7. Impact of CDT Toxin on Human Diseases.

    Science.gov (United States)

    Faïs, Tiphanie; Delmas, Julien; Serres, Arnaud; Bonnet, Richard; Dalmasso, Guillaume

    2016-07-15

    Cytolethal distending toxin (CDT) is found in Gram-negative bacteria, especially in certain Proteobacteria such as the Pasteurellaceae family, including Haemophilus ducreyi and Aggregatibacter (Actinobacillus) actinomycetemcomitans, in the Enterobacteriaceae family and the Campylobacterales order, including the Campylobacter and Helicobacter species. In vitro and in vivo studies have clearly shown that this toxin has a strong effect on cellular physiology (inflammation, immune response modulation, tissue damage). Some works even suggest a potential involvement of CDT in cancers. In this review, we will discuss these different aspects.

  8. Comparrisson of MICs of ceftioufur and other antimicrobial agents against bacterial pathogens of swine from the United States, Canada and Denmark

    DEFF Research Database (Denmark)

    Salmon, S.A.; Watts, J.L.; Case, C.A.;

    1995-01-01

    The MICs of ceftiofur and other antimicrobial agents, tested for comparison, for 515 bacterial isolates of pigs from the United States, Canada, and Denmark with various diseases were compared. The organisms tested included Actinobacillus pleuropneumoniae, Escherichia coli, Pasteurella multocida....../ml). However, this antimicrobial agent was much less active when it was tested against A. pleuropneumoniae, S. cholerae-suis, and E. coli (MIC(90)s, 16.0, >32.0, and >32.0 mu g/ml, respectively). Against the U.S. isolates of A. pleuropneumoniae and P. multocida, tilmicosin was moderately active (MIC(90)s, 4...

  9. Prevalence and distribution of principal periodontal pathogens worldwide

    DEFF Research Database (Denmark)

    Rylev, Mette; Kilian, Mogens

    2008-01-01

    to particular ethnic groups. AIM: This review analyzes to what extent observed differences in periodontal disease prevalence among ethnically or geographically distinct populations may be explained by restricted host adaptation of clones of principal periodontal pathogens. RESULTS: Carriage rates of several...... putative periodontal pathogens and particular subsets of these species vary between ethnic groups. Few of these differences can, with the limited information available, be directly related to differences in periodontal disease prevalence. Asian populations are regularly colonized with Actinobacillus...... are likely to give better insight into the aetiology of periodontal diseases....

  10. Recombinant microorganisms for increased production of organic acids

    Energy Technology Data Exchange (ETDEWEB)

    Yi, Jian; Kleff, Susanne; Guettler, Michael V

    2013-04-30

    Disclosed are recombinant microorganisms for producing organic acids. The recombinant microorganisms express a polypeptide that has the enzymatic activity of an enzyme that is utilized in the pentose phosphate cycle. The recombinant microorganism may include recombinant Actinobacillus succinogenes that has been transformed to express a Zwischenferment (Zwf) gene. The recombinant microorganisms may be useful in fermentation processes for producing organic acids such as succinic acid and lactic acid. Also disclosed are novel plasmids that are useful for transforming microorganisms to produce recombinant microorganisms that express enzymes such as Zwf.

  11. Laminaria digitata as a potential carbon source for succinic acid and bioenergy production in a biorefinery perspective

    DEFF Research Database (Denmark)

    Alvarado-Morales, Merlin; Gunnarsson, Ingólfur Bragi; Fotidis, Ioannis

    2015-01-01

    A novel biorefinery concept utilizing macroalgae Laminaria digitata to produce succinic acid, and direct the process residues for feed and energy production, is investigated in the present study. Enzymatic hydrolysis was performed at high solid loading (25% w v− 1) resulting in solubilization...... of the carbohydrates to soluble sugars, which accumulated in the liquid hydrolysate. The overall sugar recovery in the macroalgae hydrolysate was 78.23%. Actinobacillus succinogenes 130Z was able to ferment macroalgae hydrolysate to succinic acid with a yield of 86.49% (g g− 1 of total sugars) and an overall...

  12. Anti-microbial Activity of Tulsi {Ocimum Sanctum (Linn.)} Extract on a Periodontal Pathogen in Human Dental Plaque: An Invitro Study

    Science.gov (United States)

    Devaraj, C.G.; Agarwal, Payal

    2016-01-01

    Introduction Tulsi is a popular healing herb in Ayurvedic medicine. It is widely used in the treatment of several systemic diseases because of its anti-microbial property. However, studies documenting the effect of Tulsi on oral disease causing organisms are rare. Hence, an attempt was made to determine the effect of Tulsi on a periodontal microorganism in human dental plaque. Aim To determine if Ocimum sanctum (Linn.) has an anti-microbial activity (Minimum Inhibitory Concentration and zone of inhibition) against Actinobacillus actinomycetemcomitans in human dental plaque and to compare the antimicrobial activity of Ocimum sanctum(Linn.) extract with 0.2% chlorhexidine as the positive control and dimethyl sulfoxide as the negative control. Materials and Methods A lab based invitro experimental study design was adopted. Ethanolic extract of Ocimum sanctum (Linn.) was prepared by the cold extraction method. The extract was diluted with an inert solvent, dimethyl sulfoxide, to obtain ten different concentrations (1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%) of extract. Plaque sample was collected from 05 subjects diagnosed with periodontal disease. Isolation of Actinobacillus actinomycetemcomitans from plaque samples was done using Tryptic Soy Serum Bacitracin Vancomycin agar (TSBV) medium. Identification of Actinobacillus actinomycetemcomitans was done based on cultural, microscopic, biochemical characterization and multiple drug resistance patterns. Anti-microbial activity of Ocimum sanctum (Linn.) extract was tested by agar well-diffusion method against 0.2% chlorhexidine as a positive control and dimethyl sulfoxide as a negative control. The zone of inhibition was measured in millimeters using Vernier callipers. Results At the 6% w/v concentration of Ocimum sanctum (Linn.) extract, a zone of inhibition of 22 mm was obtained. This was the widest zone of inhibition observed among all the 10 different concentrations tested. The zone of inhibition for positive control

  13. Infective endocarditis due to Enterobacter cloacae resistant to third- and fourth-generation cephalosporins.

    Science.gov (United States)

    Yoshino, Yusuke; Okugawa, Shu; Kimura, Satoshi; Makita, Eiko; Seo, Kazunori; Koga, Ichiro; Matsunaga, Naohisa; Kitazawa, Takatoshi; Ota, Yasuo

    2015-04-01

    We report the case of using a long-term combination of meropenem and amikacin to treat infective endocarditis caused by Enterobacter cloacae resistant to third- and fourth-generation cephalosporins. Multi-drug resistant Gram-negative bacilli, such as the E. cloacae in our study, may become possible pathogens of infective endocarditis. Our experience with this case indicates that long-term use of a combination of β-lactam and aminoglycosides might represent a suitable management option for future infective endocarditis cases due to non-Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, Kingella spp. (HACEK group) Gram-negative bacilli such as ours.

  14. 伴放线凝聚杆菌感染性心内膜炎一例分析

    Institute of Scientific and Technical Information of China (English)

    刘薇; 云峰; 陈龙; 杨洪波; 王拓; 王晓玲; 姚瑶; 徐珊; 王玉彬; 赖惠英; 陈雪; 陶凤蓉; 艾效曼; 陈东科; 许宏涛; 胡云建

    2014-01-01

    感染性心内膜炎(infective endocarditis,IE)为细菌、真菌和其他微生物(如病毒、立克次体、衣原体、螺旋体等)直接感染而产生心瓣膜或心室壁内膜的炎症。嗜血杆菌属(Haemophilus species)、放线杆菌属(Actinobacillus actinomycetemcomitans)、心杆菌属(Cardiobacterium hominis)、艾肯菌属(Eikenella corrodens)、

  15. Aspectos recentes da patogênese e diagnóstico da pleuropneumonia suína

    Directory of Open Access Journals (Sweden)

    Vaz Clarissa Silveira Luiz

    2004-01-01

    Full Text Available A pleuropneumonia suína, causada por Actinobacillus pleuropneumoniae, é uma doença caracterizada pela apresentação fibrino-hemorrágica com pleurite adesiva. A enfermidade está presente em todos os países produtores de suínos, sendo responsável por prejuízos econômicos elevados. No Brasil e no mundo, diversos grupos vêm conduzindo estudos na busca por um melhor entendimento da doença e de sua epidemiologia. Avanços importantes foram obtidos, entre os quais a caracterização dos fatores de virulência, implicados na apresentação clínica da enfermidade; e a aplicação de novos métodos de diagnóstico. A difusão das técnicas de biologia molecular como ferramenta diagnóstica em Medicina Veterinária tem contribuindo para a identificação de Actinobacillus pleuropneumoniae. Nesta revisão, são abordados os aspectos mais recentes sobre a patogênese e o diagnóstico deste importante patógeno.

  16. Susceptibility of bacteria isolated from pigs to tiamulin and enrofloxacin metabolites

    DEFF Research Database (Denmark)

    Lykkeberg, Anne Kruse; Halling-Sørensen, Bent; Jensen, Lars Bogø

    2007-01-01

    -tiamulin (8 alpha-HTIA), and the ENR metabolites were: ciprofloxacin (CIP) and enrofloxacin N-oxide (ENR-N). Bacteria, all of porcine origin, we're selected as representatives of bacterial infections (Stap4ylococcus hyicus and Actinobacillus pleuropneumoniae), zoonotic bacteria (Campylobacter coli......:Susceptibilities to metabolites of tiamulin (TIA) and enrofloxacin (ENR) were tested using selected bacteria with previously defined minimal inhibitory concentrations,(,MIC). The TIA metabolites tested were: N-deethyl-tiamulin (I)TIA), 2 beta-hydroxy-tiamulin (2 beta-HTIA),and Sammhydroxy......) and indicator bacteria (Escherichia coli and Furthermore the effects of ithese compounds were tested on the microbial community of active sludge to test any negative effect on colony forming units,(CFU). DTIA had a potency of 12.5-50% of the potency of T1A. 2-HTIA:and 8 alpha HTIA had,potenciesless, than 1...

  17. Molecular characterization of the porcine surfactant, pulmonary-associated protein C gene

    DEFF Research Database (Denmark)

    Cirera, S.; Nygård, A.B.; Jensen, H.E.;

    2006-01-01

    The surfactant, pulmonary-associated protein C (SFTPC) is a peptide secreted by the alveolar type II pneumocytes of the lung. We have characterized the porcine SFTPC gene at genomic, transcriptional, and protein levels. The porcine SFTPC is a single-copy gene on pig chromosome 14. Two transcripts...... were found in a newborn pig lung cDNA library: a full-length clone and a clone missing exon 5. cDNA sequence comparison revealed four synonymous and two nonsynonymous substitutions and in-frame insertions at the beginning of exon 5. Comparison of the SFTPC coding region between several mammals showed......-regulated in necrotic lungs of pigs infected with Actinobacillus pleuropneumoniae. Additionally, the protein levels were also decreased or absent in the necrotic tissue....

  18. [Glanders--an eradicable disease--or a threat?].

    Science.gov (United States)

    Pospísil, L

    2001-12-01

    Glanders (malleus), attacking equids and transmissible to humans, does not occur in our geographical area any more, but world-wide eradication has not yet been achieved. Cases of glanders have been reported from India, Iraq, Mongolia and China and in 2001 also from South America. The disease is caused by Burkholderia mallei (earlied known as Bacillus, Pfeiferella, Loefflerella, Malleomyces, Actinobacillus, or Pseudomonas mallei). The continual interest of microbiologists in the causative agents indicates that glanders cannot be regarded as a closed historic episode. Occupational infections of laboratory personnel occurred during World War II and the years thereafter and the last accident was reported in May 2000. Topical problems of glanders include the development of a vaccine and antibiotic therapy tested in experimentally infected subjects.

  19. Pleuritis in slaughter pigs: relations between lung lesions and bacteriology in 10 herds with high pleuritis.

    Science.gov (United States)

    Jirawattanapong, Pichai; Stockhofe-Zurwieden, Norbert; van Leengoed, Leo; Wisselink, Henk; Raymakers, Rudolf; Cruijsen, Toine; van der Peet-Schwering, Carola; Nielen, Mirjam; van Nes, Arie

    2010-02-01

    Pleuritis in slaughter pigs has increased in recent years in the Netherlands. The aim of the present study was to determine what respiratory pathogens were involved in pleuritis. In total, lungs of 968 slaughter pigs from 10 herds with high prevalence of pleuritis were morphologically examined for size, location, and type of lesions. Moreover, histology and bacteriology were performed. Examination of gross lung lesions showed 45% pleuritis, 14% pleuropneumonia and 38% catarrhal pneumonia. Peribronchiolar cuffing was found in 61 of 142 samples. Actinobacillus pleuropneumoniae was cultured from 22 lung samples from four herds. Pasteurella multocida was cultured from 55 lung samples in eight herds. No specific pattern with respect to the causal pathogens was found. In conclusion, no single infectious cause of pleuritis was found. A variety of infectious agents combined with environmental factors should be considered as a cause of pleuritis.

  20. Doxycycline sustained release from brushite cements for the treatment of periodontal diseases.

    Science.gov (United States)

    Tamimi, Faleh; Torres, Jesús; Bettini, Ruggero; Ruggera, Francesca; Rueda, Carmen; López-Ponce, Manuel; Lopez-Cabarcos, Enrique

    2008-06-01

    Doxycycline (DOXY) is a wide spectrum antibiotic used in the treatment of dental, periodontal, and bone infections. Brushite cements are calcium phosphate biomaterials especially interesting for bone regeneration processes. In this work, we describe the preparation of a brushite cement containing DOXY and the drug release from the cement. DOXY solutions were mixed with the cement powder and after a 50% burst release in the first 12 h, a slow and controlled release was achieved over 3.5 days. The release of DOXY hyclate was controlled by both, diffusion and Ca(2+) interaction. Formation of DOXY-Ca(2+) chelates was detected in the cement structure using solid state fluorescence. The brushite cement loaded with DOXY hyclate had antibacterial activity against periodontal pathogens: Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Bacteroides frosthytus. This new biomaterial may be helpful for the treatment of periodontal diseases.

  1. 猪传染性胸膜肺炎的诊治

    Institute of Scientific and Technical Information of China (English)

    田海蓉; 张登祥

    2007-01-01

    猪传染性胸膜肺炎(Porcine Contagious pleuropneu—moniae,PCP)是由胸膜肺炎放线杆菌(Actinobacillus pleurop—neumoniae,APP)引起猪的一种呼吸系统传染病,任何年龄段猪均可发生,经空气或直接接触传播,应激可促使急性暴发,表现为典型的胸膜肺炎、鼻孔分泌物及口腔呕吐物带血。本病对养猪户造成的直接经济损失主要体现在病猪死亡损失和治疗药物费用上。

  2. Thermochemical pretreatments for enhancing succinic acid production from industrial hemp (Cannabis sativa L.)

    DEFF Research Database (Denmark)

    Gunnarsson, Ingólfur Bragi; Kuglarz, Mariusz; Karakashev, Dimitar Borisov

    2015-01-01

    The aim of this study was to develop an efficient thermochemical method for treatment of industrial hemp biomass, in order to increase its bioconversion to succinic acid. Industrial hemp was subjected to various thermochemical pretreatments using 0-3% H2SO4, NaOH or H2O2 at 121-180°C prior...... to enzymatic hydrolysis. The influence of the different pretreatments on hydrolysis and succinic acid production by Actinobacillus succinogenes 130Z was investigated in batch mode, using anaerobic bottles and bioreactors. Enzymatic hydrolysis and fermentation of hemp material pretreated with 3% H2O2 resulted...... in the highest overall sugar yield (73.5%), maximum succinic acid titer (21.9gL-1), as well as the highest succinic acid yield (83%). Results obtained clearly demonstrated the impact of different pretreatments on the bioconversion efficiency of industrial hemp into succinic acid....

  3. Periodontal pathogens in erupting third molars of periodontally healthy subjects.

    Science.gov (United States)

    Rajasuo, A; Sihvonen, O J; Peltola, M; Meurman, J H

    2007-09-01

    The presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and Tannerella forsythensis in bacteriologic samples of 5-7-mm deep mandibular third-molar pericoronal pockets was analysed by polymerase chain reaction, to test the hypothesis that these sites would harbour the bacteria. The patients were periodontally healthy 20-year-old Finnish male conscripts. Sixteen had acute pericoronitis, 28 chronic pericoronitis, and 15 were symptom-free controls. A. actinomycetemcomitans was detected in only 7% of the samples from chronic pericoronitis cases, whereas P. gingivalis was positive in 20% of the symptom-free versus 69% (P = 0.018) of the acute and 57% (P = 0.044) of the chronic cases. The percentages for P. intermedia were 93, 94 and 93%, and for T. forsythensis 47, 63 and 57%, respectively. These results confirm that, apart from A. actinomycetemcomitans, periodontopathogens are common in third-molar sites in periodontally healthy individuals.

  4. Transcriptional profiling at different sites in lungs of pigs during acute bacterial respiratory infection

    DEFF Research Database (Denmark)

    Mortensen, Shila; Skovgaard, Kerstin; Hedegaard, Jakob

    2011-01-01

    The local transcriptional response was studied in different locations of lungs from pigs experimentally infected with the respiratory pathogen Actinobacillus pleuropneumoniae serotype 5B, using porcine cDNA microarrays. This infection gives rise to well-demarcated infection loci in the lung...... of apoptosis and the complement system. Interferon-g was downregulated in both necrotic and bordering areas. Evidence of neutrophil recruitment was seen by the up-regulation of chemotactic factors for neutrophils. In conclusion, we found subsets of genes expressed at different levels in the three selected...... of induced genes as, in unaffected areas a large part of differently expressed genes were involved in systemic reactions to infections, while differently expressed genes in necrotic areas were mainly concerned with homeostasis regulation....

  5. Determination of the antibacterial activity of simvastatin against periodontal pathogens, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans: An in vitro study

    Directory of Open Access Journals (Sweden)

    Shilpa Emani

    2014-01-01

    Full Text Available Context and Objective: Statin treatment, apart from its hypolipidemic action has proven its antimicrobial activity by improving the survival rate of patients with severe systemic bacterial infections. Periodontitis is an inflammatory disorder of tooth supporting structures caused by a group of specific microorganisms. The objective of the present study was to determine the antimicrobial activity of pure simvastatin drug against the primary periodontal pathogens. Materials and Methods: Minimum inhibitory concentration (MIC was determined against Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans using serial dilution method. Results: MIC of simvastatin against P. gingivalis was 2 μg/ml and A. actinomycetemcomitans was found to be <1 μg/ml which requires further dilutions to determine the exact value. Conclusions: Data suggests a potent antimicrobial activity of simvastatin against both A. actinomycetemcomitans and P gingivalis. Hence simvastatin can be prescribed as a dual action drug in patients with both hyperlipidemia and periodontal disease.

  6. Microbial flora in orodental infections

    Directory of Open Access Journals (Sweden)

    Saini S

    2003-01-01

    Full Text Available The present study was carried out to compare the normal aerobic and anaerobic bacterial oral flora with flora from deep seated dental caries, gingivitis and adult periodontitis. All the samples belonging to both the control and study groups yielded microbes. Aerobe / Anaerobe ratio was high in normal flora (1.48 as compared to dental caries (0.9, gingivitis (0.72 and periodontitis (0.56. Ninety seven percent of orodental infections were polymicrobial and three or more microbes were found in 84% cases of study group as compared to 28% in controls. Streptococcus mutans and anaerobic lactobacilli were common in dental caries, Actinomyces and Peptostreptococcus spp. in gingivitis, Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in periodontitis.

  7. Thermochemical pretreatments for enhancing succinic acid production from industrial hemp (Cannabis sativa L.).

    Science.gov (United States)

    Gunnarsson, Ingólfur B; Kuglarz, Mariusz; Karakashev, Dimitar; Angelidaki, Irini

    2015-04-01

    The aim of this study was to develop an efficient thermochemical method for treatment of industrial hemp biomass, in order to increase its bioconversion to succinic acid. Industrial hemp was subjected to various thermochemical pretreatments using 0-3% H2SO4, NaOH or H2O2 at 121-180°C prior to enzymatic hydrolysis. The influence of the different pretreatments on hydrolysis and succinic acid production by Actinobacillus succinogenes 130Z was investigated in batch mode, using anaerobic bottles and bioreactors. Enzymatic hydrolysis and fermentation of hemp material pretreated with 3% H2O2 resulted in the highest overall sugar yield (73.5%), maximum succinic acid titer (21.9 g L(-1)), as well as the highest succinic acid yield (83%). Results obtained clearly demonstrated the impact of different pretreatments on the bioconversion efficiency of industrial hemp into succinic acid.

  8. Occurrence of antimicrobial resistance among bacterial pathogens and indicator bacteria in pigs in different European countries from year 2002 – 2004: the ARBAO-II study

    DEFF Research Database (Denmark)

    Hendriksen, Rene S.; Mevius, Dik J; Schroeter, Andreas

    2008-01-01

    of pathogenic and indicator bacteria from food animals using validated and harmonised methodologies. In this report the first data on the occurrence of antimicrobial resistance among bacteria causing infections in pigs are reported. Methods: Susceptibility data from 17,642 isolates of pathogens and indicator......Background: The project "Antibiotic resistance in bacteria of animal origin - II" (ARBAO-II) was funded by the European Union (FAIR5-QLK2-2002-01146) for the period 2003-05. The aim of this project was to establish a program for the continuous monitoring of antimicrobial susceptibility...... bacteria including Actinobacillus pleuropneumoniae, Streptococcus suis and Escherichia coli isolated from pigs were collected from fifteen European countries in 2002-2004. Results: Data for A. pleuropneumoniae from infected pigs were submitted from five countries. Most of the isolates from Denmark were...

  9. Expression of Matrix Metalloproteinase-9 and -12 in Porcine Lung Infections

    DEFF Research Database (Denmark)

    Bruun, C. S.; Leifsson, P. S.; Johansen, L. K.

    2012-01-01

    Matrix metalloproteinases (MMPs) play a variety of roles during organogenesis, in the immune response and during acute and chronic diseases as well as in tissue remodelling. During the last decade, the pig has become used increasingly as a model for human diseases; however, studies on the express...... expressions were seen in the alveolar epithelium (MMP-9) and alveolar macrophages (MMP-12). These results are in accordance with studies of human lungs....... on the expression of porcine MMPs are limited. In the present study species-specific antibodies were produced to investigate the expression of MMP-9 and MMP-12 immunohistochemically in lungs from pigs infected with Actinobacillus pleuropneumoniae, Pasteurella multocida and Staphylococcus aureus. The immunolabelling...... of lung tissues (one infected and one control pig representing each infection) was evaluated for cellular distribution and intensity, which was scored semiquantitatively. When compared with healthy, non-infected controls, the expression of both MMP-9 and MMP-12 was higher in infected lungs. The highest...

  10. Pulmonary infections in swine induce altered porcine surfactant protein D expression and localization to dendritic cells in bronchial-associated lymphoid tissue

    DEFF Research Database (Denmark)

    Sørensen, C.M.; Holmskov, U.; Aalbæk, B.;

    2005-01-01

    Surfactant protein D (SP-D) is a pattern-recognition molecule of the innate immune system that recognizes various microbial surface-specific carbohydrate and lipid patterns. In vitro data has suggested that this binding may lead to increased microbial association with macrophages and dendritic...... among pSP-D, pathogens, phagocytic cells and dendritic cells. Lung tissue was collected from experimental and natural bronchopneumonias caused by Actinobacillus pleuropneumoniae or Staphylococcus aureus, and from embolic and diffuse interstitial pneumonia, caused by Staph. aureus or Arcanobacterium......SP-D through the specialized M cells overlying (BALT). In conclusion, we have shown that pSP-D expression in the lung surfactant is induced by bacterial infection by an aerogenous route rather than by a haematogenous route, and that the protein interacts specifically with alveolar macrophages...

  11. Phylogeny of the genus Haemophilus as determined by comparison of partial infB sequences

    DEFF Research Database (Denmark)

    Hedegaard, J; Okkels, H; Bruun, B;

    2001-01-01

    A 453 bp fragment of infB, the gene encoding translation initiation factor 2, was sequenced and compared from 66 clinical isolates and type strains of Haemophilus species and related bacteria. Analysis of the partial infB sequences obtained suggested that the human isolates dependent on X and V...... factor, H. influenzae, H. haemolyticus, H. aegyptius and some cryptic genospecies of H. influenzae, were closely related to each other. H. parainfluenzae constituted a heterogeneous group within the boundaries of the genus, whereas H. aphrophilus/paraphrophilus and Actinobacillus actinomycetemcomitans...... were only remotely related to the type species of the genus Haemophilus H. parahaemolyticus and H. paraphrohaemolyticus took up an intermediary position and may not belong in the genus Haemophilus sensu stricto. Ambiguous results were obtained with seven isolates tentatively identified as H. segnis...

  12. Bases farmacomicrobiológicas del tratamiento antibiótico de las enfermedades periodontales y periimplatarias Farmacobiological concepts in the antibiotic treatment of the periodontal diseases

    Directory of Open Access Journals (Sweden)

    J. Liñares

    2003-12-01

    Full Text Available La enfermedad periodontal debe considerarse un proceso infeccioso bacteriano crónico. En su etiología, no hay una única especie bacteriana implicada, sino que podríamos considerarla como una infección polimicrobiana en la que estarían implicados diversos microorganismos. Las bacterias que se han asociado más directamente con la enfermedad periodontal son Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus y Treponema denticola. Los parámetros farmacodinámicos de los antibióticos son muy útiles a la hora de seleccionar pautas posológicas. El aumento de resistencias producido en muchos periodontopatógenos en los últimos años ha relegado a algunos antibióticos a un segundo plano. Entre la gran variedad de antibióticos utilizados, se han obtenido buenas respuestas terapéuticas con amoxicilina/ácido clavulánico, metronidazol, clindamicina, doxiciclina y las combinaciones de metronidazol más amoxicilina y metronidazol más amoxicili-na/ácido clavulánico.Periodontal disease must be considered a chronic bacterial infection. It does not appear to one single bacterial species that is uniquely involved. Rather, periodontal disease seems to be a polymicrobial infection involving several organisms. The bacteria most often associated with periodontal disease are Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus y Treponema denticola. Pharmacodynamics parameters are very useful to select dosing regimens. The increase in prevalence of resistance occurred in some periodontopathogens in the last years has pushed some antibiotics into the background. Positive responses have been reported with amoxicillin/clavulanate, metronidazole, clindamycin, doxycycline and the combination therapy metronidazole plus amoxicillin and metronidazole plus amoxicillin/clavulanate.

  13. Analysis of the presence of pathogens which predict the risk of disease at peri-implant sites through polymerase chain reaction (PCR Análise por reação em cadeia da polimerase (PCR da presença de patógenos preditores de risco em sítios periimplantares

    Directory of Open Access Journals (Sweden)

    Joely Ângela de Oliveira Leitão

    2005-03-01

    Full Text Available The presence of DNA of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in the peri-implant sulcus samples of 19 partially edentulous patients was analyzed by polymerase chain reaction (PCR and related to the depth of the peri-implant sulcus, bleeding on probing, and probable risk of disease. Ten of those patients presented a history of periodontal disease and nine of those did not. The DNA amplification of these pathogens was observed in seven samples, of which four were from patients without history of periodontal disease. The results suggest that even when significant inflammatory signs are absent the qualitative detection may indicate risk of peri-implantitis, requiring more strict postoperative control.A presença dos ADN de Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis e Prevotella intermedia em amostras coletadas de sulco periimplantar de 19 pacientes parcialmente desdentados foi analisada pela reação em cadeia da polimerase (PCR. Dentre esses 19 pacientes, dez apresentavam histórico de doença periodontal e nove não apresentavam antecedentes. Os resultados obtidos nesta análise foram relacionados com a profundidade do sulco periimplantar, o sangramento à sondagem e o provável risco de doença. Constatou-se que houve a amplificação do ADN das bactérias-alvo em sete amostras, sendo quatro de pacientes sem histórico de periodontopatia. Este resultado sugere que mesmo na ausência de sinais inflamatórios significantes, essa detecção qualitativa pode indicar risco de periimplantite, requerendo manutenção pós-operatória mais rigorosa.

  14. Optimization of Fermentation Medium for Succinic Acid Production Using Canna edulis ker Syrup%芭蕉芋糖浆发酵生产丁二酸培养基的优化

    Institute of Scientific and Technical Information of China (English)

    刘宇鹏; 陈万河; 陈梁; 方晨; 白彦兵; 孙志浩

    2011-01-01

    Canna edulis Ker syrup was firstly used to product succinic acid by Actinobacillus succinogenes CGMCC1593. It was found that the total sugar, corn steep liquid and phosphate significantly improved the production of succinic acid. The Response Surface Methodology (RSM) based on a three-factor Box-Behnken design of experiments was used to build an analytical model and finally employed as a constraint in an obvious optimization process. The optimized parameters were total sugar of 89.66 g/L, corn steep liquid of 38.55 g/L, and phosphate of 4.46 g/L. Under those optimization conditions, it was predicted that the the highest concentation of succinic acid recached 62.92 g/L.After optimization, the succinic acid production increased by 22. 7% , and the experimental data basically in line with the predicted values.%以芭蕉芋糖浆为主要原料,利用琥珀酸放线杆菌(Actinobacillus succinogenes)CCMCC1593发酵生产丁二酸.单因素试验表明,培养基中总糖、玉米浆和磷酸盐对发酵有显著影响,利用响应面分析法(RSM)对培养基成分进行优化,并建立了各因素与丁二酸产量之间的数学模型.当各参数取值分别为总糖89.66 g/L,磷酸盐4.46g/L,玉米浆38.55 g/L,丁二酸最大估计值为62.92 g/L.优化后丁二酸产量比优化前提高了22.7 %,与响应面预测极值基本相符.

  15. Antimicrobial proteins of Snail mucus (Achatina fulica against Streptococcus mutans and Aggregatibacter actinomycetemcomitans

    Directory of Open Access Journals (Sweden)

    Herluinus Mafranenda DN

    2014-03-01

    Full Text Available Background: Achasin and mytimacin-AF are proteins of snail mucus (Achatina fulica which have antimicrobial activity. Snail mucus is suspected to have other proteins which have antimicrobial activity against Streptococcus mutans and Aggregatibacter actinomycetemcomitans the oral pathologic bacteria. Purpose: The study were aimed to characterize the proteins of snail mucus (Achatina fulica that have antimicrobial activities to Streptococcus mutans and Actinobacillus actinomycetemcomitans, and to compared the antimicrobial effect of achasin and mytimacin-AF. Methods: The sample of study was the mucus of snails which were taken from Yogyakarta Province. The isolation and characterization of protein were conducted by using SDS-PAGE method, electroelution, and dialysis. Nano drop test was conducted to determine protein concentration. The sensitivity test was conducted by using dilution test, and followed by spectrophotometry and paper disc diffusion tests. Results: The study showed that proteins successfully characterized from snail mucus (Achatina fulica were proteins with molecular weights of 83.67 kDa (achasin, 50.81 kDa, 15 kDa, 11.45 kDa (full amino acid sequence of mytimacin-AF and 9.7 kDa (mytimacin-AF. Based on the dilution test, Achasin had better antimicrobial activities against Streptococcus mutans, while mytimacin-AF had better antimicrobial activities against Aggregatibacter actinomycetemcomitans. But the paper disc diffusion test result showed that Achasin had antimicrobial activities against Streptococcus mutans and Aggregatibacter actinomycetemcomitans, while mytimacin-AF had no antimicrobial activities. Conclusion: The proteins with molecular weights of 50.81 kDa, 15 kDa, 11.45 kDa were considered as new antimicrobial proteins isolated from snail mucus. Achasin, had better antimicrobial activities against Streptococcus mutans, while mytimacin-AF had better antimicrobial activities against Aggregatibacter actinomycetemcomitans

  16. Survey of pleuritis and pulmonary lesions in pigs at abattoir with a focus on the extent of the condition and herd risk factors.

    Science.gov (United States)

    Merialdi, G; Dottori, M; Bonilauri, P; Luppi, A; Gozio, S; Pozzi, P; Spaggiari, B; Martelli, P

    2012-07-01

    Cranioventral pulmonary consolidation (enzootic pneumonia-like lesions) and chronic pleuritis (CP) are common findings in slaughtered pigs. Pleural lesions involving dorsocaudal lobes are suggestive of pleuropneumonia due to Actinobacillus pleuropneumoniae. In this report the results of an abattoir survey of pleuritis and pulmonary lesions in pigs is presented with a focus on herd risk factors. A total of 4889 animals, ranging in age from 9 to 10 months, from 48 batches of pigs belonging to an equal number of herds, were included in the study. Bronchopneumonic lesions suggestive of enzootic pneumonia (EP-like lesions) were detected in 46.4% of the examined lungs. The EP-like lesion average value for all lungs was 1.03 (95% CI 0.98-1.08), ranging from 0.17 to 2.56 among the 48 batches; 47.5% of lungs showed chronic pleuritis. Dorsocaudal pleuritis suggestive of recovered pleuropneumonia (SPES score ≥2) was found in 25.1% of the lungs. The mean SPES (slaughterhouse pleuritis evaluation system) value of the overall 4889 lungs was 0.83 (95% CI 0.78-0.86). The mean SPES value of the batches ranged from 0.04 to 1.87. The mean Actinobacillus pleuropneumoniae index of all studied batches was 0.61 (95% CI 0.51-0.71), ranging from 0 to 1.84. Blood samples were collected from each herd to evaluate antibody titres to Mycoplasma hyopneumoniae, A. pleuropneumoniae, Aujeszky's disease virus, porcine reproductive and respiratory syndrome virus (PRRSV), and swine influenza virus. Herd characteristics were recorded using a questionnaire given to the farmers. A multivariable analysis was conducted to identify risk factors for pleuritis and EP-like lesions. High dorsocaudal pleuritis was associated with A. pleuropneumoniae seroprevalence and history of A. pleuropneumoniae isolation from pneumonic lungs of dead animals. Vaccination of weaners at 3-5 weeks of age against PRRS using a modified live vaccine was associated with a reduction in the percentage of cranioventral pulmonary

  17. 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

  18. The isolation and identification of intestinal bacteria from larval Holotrichia parallela Motschalsky ( Coleoptera: Scarabaeidae)%暗黑鳃金龟甲幼虫肠道细菌分离及鉴定

    Institute of Scientific and Technical Information of China (English)

    朱琳; 刘玉升; 刘宁; 高尚坤; 郑继法

    2012-01-01

    目的 从微生态学角度研究暗黑鳃金龟甲幼虫营养生理活动,探讨其肠道菌群的构成,为其资源开发及生物防治提供理论依据.方法 按传统分离方法,从暗黑鳃金龟甲幼虫肠道环境中分离纯化获得10个细菌菌株,对其菌体形态、染色反应、培养性状、生理生化反应进行了系统研究.结果 研究结果表明,上述10个细菌菌株分别属于鲁氏耶尔森菌(Yersinia.ruckeri)、侧胞芽胞杆菌(Bacillus laterosporus)、坚强芽胞杆菌(Bacillus firmus)、放线杆菌属(Actinobacillus)、飞虫杀雄菌(Arsenophonus nasoniae)、不动杆菌属(Acinetobacter)、气单胞菌(Aeromonas)、沙门菌属(Salmonella)、短芽胞杆菌(Bacillus brevis)、变形菌属(Proteus).结论 通过对暗黑鳃金龟甲幼虫肠道细菌的鉴定,其肠道细菌在培养性状、生理性状、生理生化测定等方面存在较多差异.%Objective To discuss the composition of intestinal flora of Holotrichia parallela Motschalsky and provide theoretical foundation for its exploitation and biological control. Method Ten bacteria strains were isolated from the intestine of the larval Holotrichia parallela Motschalsky by traditional isolation and purification method. Their morphology, staining reactions , cultural characteristics, physiological and biochemical reactions were studied systematically. Result The 10 bacteria strains belong to Yersinia. Ruckeri, Bacillus laterosporus, Bacillus firmus, Actinobacillus, Arsenophonus nasoniae, Acinetobacter, Aeromonas, Salmonella, Bacillus brevis and Proteus. Conclusion The intestinal bacterial flora of the larval Holotrichia parallela Motschalsky have many different characteristics to a certain extent.

  19. Fermentation of succinic acid using cellobiose from cellulose hydrolysates%利用纤维素水解液中的纤维二糖发酵制备丁二酸

    Institute of Scientific and Technical Information of China (English)

    徐蓉; 奚永兰; 张九花; 戴文宇; 万月佳; 陈可泉; 姜岷

    2013-01-01

    Cellobiose was often found in cellulose hydrolysates. In this study, the ability to use cellobiose to produce succinic acid by A.succinogenes NJ113 and using cellobiose prepared from sugarcane bagasse cellulose hydrolysates as a carbon source to produce succinic acid were investigated. A final succinic acid concentration of 23.51 g/L with a yield of 67.17% was achieved from an initial cellobiose concentration of 35 g/L via batch fermentation. In batch fermentation with 18 g/L of cellobiose and 17 g/L of other sugars from sugarcane bagasse cellulose hydrolysates, a succinic acid concentration of 20.00 g/L was obtained, with a yield of 64.73%. This study suggested that A. succinogenes had a strong ability to convert cellobiose into succinic acid and cellobiose from cellulose hydrolysate could be a potential carbon source for economical and efficient succinic acid production by A. succinogenes.%纤维素水解液中通常含有纤维二糖.本文考察了Actinobacillus succinogenes NJ 113利用纤维二糖厌氧发酵生产丁二酸的能力,并利用蔗渣纤维素制备纤维二糖作为碳源用于厌氧发酵生产丁二酸.3L发酵罐厌氧发酵结果显示:以35 g/L纤维二糖作为碳源发酵制备丁二酸,其产量为23.51 g/L,产率达到67.17%;用含有18g/L纤维二糖和17 g/L其它糖类的蔗渣纤维素水解液作为碳源发酵制备丁二酸,丁二酸的产量和产率分别为20.00 g/L和64.73%.因此,Actinobacillus succinogenes NJ 113具有较强的利用纤维二糖生产丁二酸的能力,而且利用废弃的纤维素制备纤维二糖作为碳源高效、经济地发酵制备丁二酸具有可行性.

  20. Ischemic Stroke and Septic Shock After Subacute Endocarditis Caused by Haemophilus parainfluenzae: Case Report

    Science.gov (United States)

    Menegueti, Mayra Goncalves; Machado-Viana, Jaciara; Gaspar, Gilberto Gambero; Nicolini, Edson Antonio; Basile-Filho, Anibal; Auxiliadora-Martins, Maria

    2017-01-01

    Haemophilus parainfluenzae, which belongs to the HACEK (Haemophilus ssp, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, and Kingella kingae) group, is a rare cause of subacute endocarditis and may lead to ischemic stroke. A 65-year-old female patient previously diagnosed with rheumatic valve disease was submitted to surgical mitral valve repair in 1996. Physical examination did not reveal any murmurs; physical examination of the lungs and abdomen was normal. The patient was admitted to hospital with progressive dyspnea, dry cough, and fever. Transesophageal echocardiogram revealed an approximately 8-mm filamentous image with chaotic motion in the ventricular face of the anterior mitral valve leaflet compatible with vegetation. Treatment with ceftriaxone and gentamicin was initiated. Haemophilus parainfluenzae grew in five blood culture samples. Along the hospital stay, the patient’s level of consciousness decreased, and she was diagnosed with ischemic stroke of cardioembolic etiology. The patient developed septic shock refractory to the prescribed treatment and died 12 days after admission. Even though the patient started being treated for endocarditis before the infectious agent was identified, the prompt use of antimicrobials hindered the growth of Haemophilus parainfluenzae and made its isolation difficult. PMID:27924179

  1. Cardiobacterium hominis Endocarditis: A Case Report and Review of the Literature

    Directory of Open Access Journals (Sweden)

    Andrew Walkty

    2005-01-01

    Full Text Available The present case report describes the clinical course of a patient who presented with Cardiobacterium hominis endocarditis. A review of the literature follows the case presentation. C hominis, a fastidious Gram-negative bacillus, is a member of the HACEK group of microorganisms (Haemophilus species, Actinobacillus actinomycetemcomitans, C hominis, Eikenella corrodens and Kingella kingae. Endocarditis caused by C hominis is uncommon and generally follows a subacute course. Patients may present with constitutional symptoms, symptoms related to valvular destruction or symptoms secondary to embolic events. Diagnosis requires identification of the pathogen from blood or vegetation by either culture or molecular techniques. Blood cultures may require prolonged incubation, highlighting the importance of incubating blood cultures for at least two to three weeks in patients with suspected endocarditis. In the past, C hominis was generally sensitive to penicillin. However, reports of beta-lactamase-producing C hominis have appeared in the literature over the past decade. The current recommendation for first-line treatment is a third-generation cephalosporin (ceftriaxone for four weeks (six weeks if a prosthetic valve is in place.

  2. Daya Antibakteri Albumen Telur Ayam Kampung (Gallus Domesticus dan Ayam Kate (Gallus Bantam terhadap Spesies Bakteri Coliform Fekal pada Cangkang Telur

    Directory of Open Access Journals (Sweden)

    Vera Pramesti Wijaya

    2014-06-01

    Full Text Available Penelitian ini bertujuan mengidentifikasi spesies-spesies bakteri koliform fekal yang terdapat pada cangkang telur ayam kampung dan ayam kate dan menganalisis pengaruh albumen telur ayam kampung dan telur ayam kate terhadap penghambatan pertumbuhan spesies-spesies bakteri koliform fekal yang terdapat pada cangkang telur ayam. Penelitian ini merupakan penelitian ekperimen dengan variabel bebas berupa albumen telur ayam kampung dan kate. Variabel terikat berupa zona hambat pertumbuhan bakteri koliform fekal. Pengujian dilakukan dengan metode difusi agar. Pengujian daya antibakteri albumen telur ayam kampung dan ayam kate dilakukan dengan mengukur diameter zona hambat pertumbuhan koloni tiap spesies bakteri koliform fekal pada medium Nutrien Agar. Data pe-nelitian adalah data hasil pengukuran diameter zona hambat pertumbuhan spesies-spesies bakteri koliform fekal. Hasil penelitian dianalisis menggunakan analisis varian tunggal, dan dilanjutkan dengan uji BNT 1%. Hasil dari penelitian adalah: (1 spesies-spesies bakteri koliform fekal yang terdapat pa-da cangkang telur ayam kampung dan telur ayam yaitu Actinobacillus sp., Serratia liquefaciens, Klebsiella ozaenae, dan Escherichia vulneris; dan (2 ada pengaruh perbedaan albumen telur ayam kampung dan telur ayam kate terhadap daya hambat pertumbuhan spesies-spesies bakteri koliform fekal yang terdapat pada cangkang telur ayam.Kata kunci: albumen telur, ayam kampung, ayam kate, daya antibakteri, bakteri koliform fekal

  3. Characterization and antibacterial performance of ZrCN/amorphous carbon coatings deposited on titanium implants

    Energy Technology Data Exchange (ETDEWEB)

    Lai, Chih-Ho [School of Medicine, China Medical University, Taichung, 404 Taiwan (China); Chang, Yin-Yu, E-mail: yinyu@mail2000.com.tw [Department of Mechanical and Computer-Aided Engineering, National Formosa University, Yunlin, Taiwan (China); Huang, Heng-Li [School of Dentistry, China Medical University, Taichung, Taiwan (China); Kao, Ho-Yi [Department of Materials Science and Engineering, Mingdao University, Changhua, Taiwan (China)

    2011-12-30

    Titanium (Ti)-based materials have been used for dental/orthopedic implants due to their excellent biological compatibility, superior mechanical strength and high corrosion resistance. The osseointegration of Ti implants is related to their composition and surface treatment. Better biocompatibility and anti-bacterial performances of Ti implant are beneficial for the osseointegration and for avoiding the infection after implantation surgery. In this study, nanocomposite ZrCN/amorphous carbon (a-C) coatings with different carbon contents were deposited on a bio-grade pure Ti implant material. A cathodic-arc evaporation system with plasma enhanced duct equipment was used for the deposition of ZrCN/a-C coatings. Reactive gas (N{sub 2}) and C{sub 2}H{sub 2} activated by the zirconium plasma in the evaporation process were used to deposit the ZrCN/a-C coatings. To verify the susceptibility of implant surface to bacterial adhesion, Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans), one of the major pathogen frequently found in the dental implant-associated infections, was chosen for in vitro anti-bacterial analyses. In addition, the biocompatibility of human gingival fibroblast (HGF) cells on coatings was also evaluated by a cell proliferation assay. The results suggested that the ZrCN/a-C coatings with carbon content higher than 12.7 at.% can improve antibacterial performance with excellent HGF cell compatibility as well.

  4. The antibacterial activities of aditoprim and its efficacy in the treatment of swine streptococcosis

    Science.gov (United States)

    Cheng, Guyue; Xu, Yamei; Zhu, Xudong; Xie, Shuyu; Wang, Liye; Huang, Lingli; Hao, Haihong; Liu, Zhenli; Pan, Yuanhu; Chen, Dongmei; Wang, Yulian; Yuan, Zonghui

    2017-01-01

    Aditoprim (ADP) has potential use as an antimicrobial agent in animals. However, its pharmacodynamic properties have not been systematically studied yet. In this study, the in vitro antibacterial activities of ADP and its main metabolites were assayed, and the in vivo antibacterial efficacy of ADP for the treatment of swine streptococcosis was evaluated. It was shown that Salmonella and Streptococcus from swine, Escherichia coli and Salmonella from chickens, E. coli, Streptococcus, Mannheimia, Pasteurella from calves, Streptococcus and Mannheimia from sheep, and E. coli, Flavobacterium columnare, Acinetobacter baumannii and Yersinia ruckeri from fishes were highly susceptible to ADP. Haemophilus parasuis from swine, Staphylococcus aureus, Aeromonas punctate, Mycobacterium tuberculosis, Streptococcus agalactiae from fishes, and Klebsiella from calves and sheep showed moderate susceptibility to ADP, whereas E. coli, Actinobacillus pleuropneumonia, Pasteurella, S. aureus, Clostridium perfringens from swine, S. aureus, C. perfringens from chickens, and S. aureus from calves were resistant to ADP. The main metabolites of ADP showed equal activity to that of their parent compound, and the prevention and therapeutic dosages of ADP recommended for swine streptococcosis were 10 and 20~40 mg/kg b.w., respectively. This study firstly showed that ADP had strong antibacterial activity and had potential to be used as a single drug in the treatment of bacterial infectious diseases. PMID:28145487

  5. Comparative cytotoxicity of periodontal bacteria

    Energy Technology Data Exchange (ETDEWEB)

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

    1988-11-01

    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.

  6. Detection of putative periodontal pathogens in non-insulin-dependent diabetes mellitus and non-diabetes mellitus by polymerase chain reaction.

    Science.gov (United States)

    Yuan, K; Chang, C J; Hsu, P C; Sun, H S; Tseng, C C; Wang, J R

    2001-02-01

    It has been assumed that there is a relationship between periodontal diseases and diabetes mellitus, however the putative periodontal microorganisms in non-diabetes mellitus (non-DM) individuals and non-insulin-dependent diabetes mellitus (NIDDM) patients have not been well studied. In this study, the detection rates of 5 putative periodontal pathogens: Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Eikenella corrodens, Treponema denticola, and Candida albicans by polymerase chain reaction (PCR) between NIDDM and non-DM adults were compared. A total of 246 adults were randomly recruited and periodontal parameters including: plaque index (P1I), gingival index (GI), probing depth (PD) and attachment level (AL) were recorded. Subgingival plaque samples were collected by sterile curettes from the most diseased and healthy sites based on PD and AL. The differences in periodontal parameters and microbiological data in healthy and diseased sites between non-DM and NIDDM patients were compared by chi-square analysis. The results showed no significant differences in age, gender, GI, P1I, PD, and prevalence of the 5 microorganisms between the NIDDM and the non-diabetic groups. However, except for A. actinomycetemcomitans, the prevalence of the periodontal microorganisms tested was significantly higher (p albicans may play important roles in the periodontitis of both NIDDM and non-DM individuals, however the etiology of periodontitis in both groups may not be different from each other.

  7. Clinical, laboratory, and immunological studies of a family with a high prevalence of generalized prepubertal and juvenile periodontitis.

    Science.gov (United States)

    López, N J

    1992-05-01

    A study of a consanguineous family with a high prevalence of localized juvenile periodontitis (LJP) and generalized prepubertal periodontitis (GPP) was done over a 7-and-a-half year period. The parents had adult periodontitis, while 2 daughters, aged 13 and 15, had LJP. Furthermore, 2 other daughters, ages 14 and 10, and a son, aged 9, were affected by GPP. Two remaining siblings were not affected. Clinical and radiographic examinations on all family members were done, and chemotaxis for blood neutrophils was assessed. Laboratory tests, immunological examinations, and evaluation for neutrophil functions were done on the GPP patients. Microbiological cultures were performed on 2 of the GPP patients, as well as in the mother. The mother, the 2 LJP patients, and 1 of the unaffected siblings had depressed PMN chemotaxis. The other family members, including the 3 GPP patients, had normal PMN chemotaxis. GPP patients did not have any systemic disease, and evidence of major defects in the immunological functions was not detected. LJP patients were successfully treated with root planing, subgingival curettage, and tetracycline therapy. Intensive periodontal therapy, combined with systemic administration of antibiotics, was not effective in halting periodontal tissue destruction in the 3 GPP patients. Results indicate that the underlying cause of GPP is not always related to leukocyte dysfunction. Since Actinobacillus actinomycetemcomitans was the most frequent pathogen found in subgingival microflora of 2 of the GPP patients, it is assumed that it may play a key role in the etiology of GPP.

  8. Rapid adaptation drives invasion of airway donor microbiota by Pseudomonas after lung transplantation

    Science.gov (United States)

    Beaume, M.; Köhler, T.; Greub, G.; Manuel, O.; Aubert, J-D.; Baerlocher, L.; Farinelli, L.; Buckling, A.; van Delden, C.; Achermann, Rita; Amico, Patrizia; Baumann, Philippe; Beldi, Guido; Benden, Christian; Berger, Christoph; Binet, Isabelle; Bochud, Pierre-Yves; Boely, Elsa; Bucher, Heiner; Bühler, Leo; Carell, Thierry; Catana, Emmanuelle; Chalandon, Yves; Geest, Sabina de; Rougemont, Olivier de; Dickenmann, Michael; Duchosal, Michel; Fehr, Thomas; Ferrari-Lacraz, Sylvie; Garzoni, Christian; Soccal, Paola Gasche; Giostra, Emiliano; Golshayan, Déla; Good, Daniel; Hadaya, Karine; Halter, Jörg; Heim, Dominik; Hess, Christoph; Hillinger, Sven; Hirsch, Hans H.; Hofbauer, Günther; Huynh-Do, Uyen; Immer, Franz; Klaghofer, Richard; Koller, Michael; Laesser, Bettina; Lehmann, Roger; Lovis, Christian; Marti, Hans-Peter; Martin, Pierre Yves; Martinolli, Luca; Meylan, Pascal; Mohacsi, Paul; Morard, Isabelle; Morel, Philippe; Mueller, Ulrike; Mueller, Nicolas J; Mueller-McKenna, Helen; Müller, Antonia; Müller, Thomas; Müllhaupt, Beat; Nadal, David; Pascual, Manuel; Passweg, Jakob; Ziegler, Chantal Piot; Rick, Juliane; Roosnek, Eddy; Rosselet, Anne; Rothlin, Silvia; Ruschitzka, Frank; Schanz, Urs; Schaub, Stefan; Seiler, Christian; Stampf, Susanne; Steiger, Jürg; Stirnimann, Guido; Toso, Christian; Tsinalis, Dimitri; Venetz, Jean-Pierre; Villard, Jean; Wick, Madeleine; Wilhelm, Markus; Yerly, Patrick

    2017-01-01

    In cystic fibrosis (CF) patients, chronic airway infection by Pseudomonas leads to progressive lung destruction ultimately requiring lung transplantation (LT). Following LT, CF-adapted Pseudomonas strains, potentially originating from the sinuses, may seed the allograft leading to infections and reduced allograft survival. We investigated whether CF-adapted Pseudomonas populations invade the donor microbiota and adapt to the non-CF allograft. We collected sequential Pseudomonas isolates and airway samples from a CF-lung transplant recipient during two years, and followed the dynamics of the microbiota and Pseudomonas populations. We show that Pseudomonas invaded the host microbiota within three days post-LT, in association with a reduction in richness and diversity. A dominant mucoid and hypermutator mutL lineage was replaced after 11 days by non-mucoid strains. Despite antibiotic therapy, Pseudomonas dominated the allograft microbiota until day 95. We observed positive selection of pre-LT variants and the appearance of novel mutations. Phenotypic adaptation resulted in increased biofilm formation and swimming motility capacities. Pseudomonas was replaced after 95 days by a microbiota dominated by Actinobacillus. In conclusion, mucoid Pseudomonas adapted to the CF-lung remained able to invade the allograft. Selection of both pre-existing non-mucoid subpopulations and of novel phenotypic traits suggests rapid adaptation of Pseudomonas to the non-CF allograft. PMID:28094327

  9. A microbiological study of Papillon-Lefévre syndrome in two patients

    Science.gov (United States)

    Robertson, K; Drucker, D; James, J; Blinkhorn, A; Hamlet, S; Bird, P

    2001-01-01

    Aim—To analyse the microflora of subgingival plaque from patients with Papillon-Lefévre syndrome (PLS), which is a very rare disease characterised by palmar-plantar hyperkeratosis with precocious periodontal destruction. Methods—Bacterial isolates were identified using a combination of commercial identification kits, traditional laboratory tests, and gas liquid chromatography. Some isolates were also subjected to partial 16S rDNA sequencing. Plaque samples were also assayed for the presence of Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans in a quantitative enzyme linked immunosorbent assay (ELISA) using monoclonal antibodies. Results—The culture results showed that most isolates were capnophilic and facultatively anaerobic species—mainly Capnocytophaga spp and Streptococcus spp. The latter included S constellatus, S oralis, and S sanguis. Other facultative bacteria belonged to the genera gemella, kingella, leuconostoc, and stomatococcus. The aerobic bacteria isolated were species of neisseria and bacillus. Anaerobic species included Prevotella intermedia, P melaninogenica, and P nigrescens, as well as Peptostreptococcus spp. ELISA detected P gingivalis in one patient in all sites sampled, whereas A actinomycetemcomitans was detected in only one site from the other patient. Prevotella intermedia was present in low numbers. Conclusions—Patients with PLS have a very complex subgingival flora including recognised periodontal pathogens. However, no particular periodontopathogen is invariably associated with PLS. Key Words: Papillon-Lefévre syndrome • periodontopathogens PMID:11328836

  10. [Antimicrobial effects of tea tree oil (Melaleuca alternifolia) on oral microorganisms].

    Science.gov (United States)

    Kulik, E; Lenkeit, K; Meyer, J

    2000-01-01

    The essential oil of Melaleuca alternifolia (tea tree oil) exhibits antimicrobial activity against a wide range of Gram-positive and Gram-negative bacteria, yeasts and fungi. In this study the bacteriostatic and bacteriocidal/fungicidal activity of a tea tree oil solution, of a new tea tree oil (Tebodont) and the respective placebo-gel, of a chlorhexidindigluconate-solution and of PlakOut was tested in vitro against ten different oral microorganisms. Minimum inhibitory concentrations were in the range from 0.0293% to 1.25% for the tea tree oil solution and from 0.0082% to 1.25% for the tea tree oil gel. The values for minimum bacteriocidal/fungicidal concentrations were in the range from 0.0521% to 2.5% for the tea tree oil solution and from tea tree oil gel. The most susceptible microorganisms were Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis, whereas Streptococcus mutans and Prevotella intermedia were the least susceptible ones. Both for the chlorhexidindigluconate solution and for PlakOut the values for the minimal inhibitory concentration and for the minimal cidal concentration were between <0.0002% and 0.0125%.

  11. Cytocompatibility and antibacterial properties of zirconia coatings with different silver contents on titanium

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Heng-Li [School of Dentistry, China Medical University, Taichung 404, Taiwan (China); Chang, Yin-Yu, E-mail: yinyu@mail2000.com.tw [Department of Mechanical and Computer-Aided Engineering, National Formosa University, Yunlin 632, Taiwan (China); Chen, Ya-Chi [Department of Materials Science and Engineering, Mingdao University, Changhua 52345, Taiwan (China); Lai, Chih-Ho [School of Medicine, China Medical University, Taichung 404, Taiwan (China); Chen, Michael Y.C. [School of Dentistry, China Medical University, Taichung 404, Taiwan (China); Division of Oral and Maxillofacial Surgery, China Medical University Hospital, Taichung 404, Taiwan (China)

    2013-12-31

    This study used a twin-gun magnetron sputtering system to deposit ZrO{sub 2}-silver (Ag) coatings on biograde pure-titanium implant materials, and the Ag content in the deposited coatings was controlled by the magnetron power. The films were then annealed using rapid thermal annealing at 350 °C for 2 min to induce the nucleation and growth of nanoparticles on the film surface. Staphylococcus aureus (S. aureus) and Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) were used for in vitro antibacterial analyses. The cytocompatibility, mRNA expression, and adhesive morphology of human gingival fibroblast (HGF) cells on the coatings were also determined. The obtained results suggest that ZrO{sub 2}-Ag composite coatings containing less than 10.6 at.% Ag show hydrophobicity, good viability and proliferation of HGF cells, and antibacterial effects on S. aureus and A. actinomycetemcomitans. Moreover, the antibacterial performance of ZrO{sub 2}-Ag coatings is superior to that pure-titanium whilst maintaining biological compatibility. - Highlights: • The annealed ZrO{sub 2}-Ag coatings showed a tetragonal-and-monoclinic structure. • Nanoparticles were well distributed in the annealed ZrO{sub 2}-Ag composite coatings. • The ZrO{sub 2}-Ag coated Ti showed hydrophobic feature. • The ZrO{sub 2}-Ag showed good antibacterial performance. • The ZrO{sub 2}-Ag showed good human gingival fibroblast cell viability.

  12. Drug resistance of bacterial dental biofilm and the potential use of natural compounds as alternative for prevention and treatment.

    Science.gov (United States)

    Kouidhi, Bochra; Al Qurashi, Yasir Mohammed A; Chaieb, Kamel

    2015-03-01

    Oral diseases, such as dental caries and periodontal disease are directly linked with the ability of bacteria to form biofilm. The development of dental caries involves acidogenic and aciduric Gram-positive bacteria colonizing the supragingival biofilm (Streptococcus, Lactobacillus and Actinomycetes). Periodontal diseases have been linked to anaerobic Gram-negative bacteria forming a subgingival plaque (Porphyromonas gingivalis, Actinobacillus, Prevotella and Fusobacterium). Cells embedded in biofilm are up to 1000-fold more resistant to antibiotics compared to their planctonic ones. Several mechanisms have been proposed to explain biofilms drug resistance. Given the increased bacterial resistance to antibiotics currently used in dentistry, a great importance is given to natural compounds for the prevention of oral bacterial growth, adhesion and colonization. Over the past decade, interest in drugs derived from medicinal plants has markedly increased. It has been well documented that medicinal plants and natural compounds confer considerable antibacterial activity against various microorganisms including cariogenic and periodontal pathogens. This paper provides a review of the literature focusing on the studies on (i) biofilm in the oral cavity, (ii) drug resistance of bacterial biofilm and (iii) the potential use of plant extracts, essential oils and natural compounds as biofilm preventive agents in dentistry, involving their origin and their mechanism of biofilm inhibition.

  13. Influencia del estrógeno en la enfermedad periodontal: revisión de literatura

    Directory of Open Access Journals (Sweden)

    V.F.C. Pazmino

    2015-08-01

    Full Text Available La etiología de la enfermedad periodontal está bien definida, dentro de los agentes etiológicos que la causan podemos citar algunos microorganismos subgingivales como: Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, Actinobacillus actinomycetemcomitans y espiroquetas. La susceptibilidad del huésped a estos agentes bacterianos también tiene un papel importante dentro del progreso y prevalencia de la enfermedad periodontal. Dentro de los factores de riesgo asociados con la enfermedad periodontal tenemos: el aumento de edad, presencia de biofilm dental o placa bacteriana patogénica, alteración del estado inmunológico, deficiencia nutricional, el uso de medicamentos como corticoides, género, estrés, tabaco, factores genéticos y condiciones sistémicas, dentro de estas se incluyen alteraciones de neutrófilos, diabetes, embarazo, osteoporosis y hormonales. La enfermedad periodontal puede ser exacerbada por determinados factores sistémicos como la deficiencia o aumento de hormonas como estrógeno y progesterona. La presente revisión de literatura tiene por objeto dilucidar la influencia del estrógeno en la enfermedad periodontal.

  14. Staphylococcus epidermidis Biofilms: Functional Molecules, Relation to Virulence, and Vaccine Potential

    Science.gov (United States)

    Mack, Dietrich; Davies, Angharad P.; Harris, Llinos G.; Knobloch, Johannes K. M.; Rohde, Holger

    Medical device-associated infections, most frequently caused by Staphylococcus epidermidis and Staphylococcus aureus, are of increasing importance in modern medicine. The formation of adherent, multilayered bacterial biofilms is crucial in the pathogenesis of these infections. Polysaccharide intercellular adhesin (PIA), a homoglycan of β-1,6-linked 2-acetamido-2-deoxy-d-glucopyranosyl residues, of which about 15% are non-N-acetylated, is central to biofilm accumulation in staphylococci. It transpires that polysaccharides - structurally very similar to PIA - are also key to biofilm formation in a number of other organisms including the important human pathogens Escherichia coli, Aggregatibacter (Actinobacillus) actinomycetemcomitans, Yersinia pestis, and Bordetella spp. Apparently, synthesis of PIA and related polysaccharides is a general feature important for biofilm formation in diverse bacterial genera. Current knowledge about the structure and biosynthesis of PIA and related polysaccharides is reviewed. Additionally, information on their role in pathogenesis of biomaterial-related and other type of infections and the potential use of PIA and related compounds for prevention of infection is evaluated.

  15. Multi-antibiotic resistant bacteria in frozen food (ready to cook food) of animal origin sold in Dhaka, Bangladesh

    Institute of Scientific and Technical Information of China (English)

    Fouzia Sultana; Kamrunnahar; Hafsa Afroz; Afroz Jahan; Md Fakruddin; Suvamoy Datta

    2014-01-01

    Objective: To investigate the bacterial load and antibiotic resistance pattern of bacterial isolates obtained from (ready to cook) frozen food samples of animal origin in Dhaka, Bangladesh. Methods: A total of 20 samples of frozen ready to cook food of animal origin were purchased from different separate grocery stores in Dhaka, Bangladesh. Bacteria were isolated and identified based on the basis of biochemical properties. Results: A total of 57 isolates has been isolated from 20 samples, of them 35.08% were Gram positive and 64.92% were Gram negative organisms. Highest percentages of isolated organisms were Staphylococcocus spp. (24.56%), Alcaligene spp. (17.54%), Klebshiella spp. (12.28%) and the lowest percentages of organisms were Enterococcus spp., Actinobacillus spp. and Proteus spp. Antibiogram results clearly showed that levofloxacin and imipenem were the most effective drug against the isolates. The less effective antibiotics were chloramphenicol and nalidixic acid and resistance was highest against ciprofloxacin. The most contaminated food was chicken nuggets. Conclusions: This type of frozen food contaminated with multi-antibiotic resistant microorganisms can be potential vehicles for transmitting food-borne diseases.

  16. The Isolation of Pathogenic Microorganisms from Imported Red Wine%进口红葡萄酒中病原微生物的检测

    Institute of Scientific and Technical Information of China (English)

    金娜; 郑文陆; 刘飞兰; 陈琳

    2014-01-01

    采用GB4789.2-2010进行菌落总数的测定,将菌落总数超标的菌落接种至营养肉汤进行增菌,接种选择性平板后挑取可疑菌落按VITEK操作手册对病原微生物进行鉴定。结果表明:从进口红葡萄酒中检出粪肠球菌和尿放线杆菌。因此,检验检疫机构在日常工作中应充分重视对低风险进口食品的检验把关,保证进口产品的质量安全,保护消费者健康。%The sample was studied according to GB4789.2-2010. While aerobic bacterial count exceeded standard of nutritional agar, the bacteria were inoculated into broth to enrich. VITEK Manual was used to identify and validate double strains. The results showed that Enterococcus faecalis and Actinobacillus ureae were simultaneously isolated from imported red wine. So, it is necessary to pay more attention to the supervision of low risk imported foods to ensure the food safety.

  17. Endocardite infecciosa causada por Eikenella corrodens

    Directory of Open Access Journals (Sweden)

    Cardoso Juliano Novaes

    2005-01-01

    Full Text Available Os microorganismos do grupo HACEK (Haemophilus spp, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens e Kingella kingae são responsáveis por 3% dos casos de endocardites. Eles apresentam propriedades clínicas e microbiológicas semelhantes entre si: são bacilos gram-negativos, isolados mais facilmente em meios aeróbicos, suas culturas necessitam de tempo prolongado de incubação para crescimento (média 3,3 dias e podem ser considerados como parte da flora normal do trato respiratório superior e da orofaringe1,2. Algumas características foram identificadas nas endocardites por esses agentes, como o quadro clínico insidioso¹, diagnóstico difícil pela natureza fastidiosa e culturas negativas3,4. A endocardite por Eikenella corrodens foi descrita pela primeira vez em 1972(5 e continua sendo um agente etiológico raro. Relatamos o caso de uma paciente com valva nativa que apresentou endocardite infecciosa causada por Eikenella corrodens.

  18. Exposure of feral swine (Sus scrofa) in the United States to selected pathogens.

    Science.gov (United States)

    Baroch, John A; Gagnon, Carl A; Lacouture, Sonia; Gottschalk, Marcelo

    2015-01-01

    Feral swine (Sus scrofa) are widely distributed in the United States. In 2011 and 2012, serum samples and tonsils were recovered from 162 and 37 feral swine, respectively, in the US to evaluate exposure to important swine endemic pathogens. Antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) were found in 2.5% and 25.3% of tested sera, respectively. Positive serological reactions against Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae have been detected in 19.7% and 69.7% of animals. More than 15% of animals presented antibodies against these 2 pathogens simultaneously. Most animals were also seropositive for Lawsonia intracellularis. Feral swine can also be involved in transmission of zoonotic agents. Almost 50% of animals possessed antibodies against Salmonella. In addition, 94.4% of animals were carriers of Streptococcus suis in their tonsils. In conclusion, feral swine may be considered as a potential reservoir for different endemic diseases in domestic pigs, as well as for important zoonotic agents.

  19. Enhanced succinic acid production from corncob hydrolysate by microbial electrolysis cells.

    Science.gov (United States)

    Zhao, Yan; Cao, Weijia; Wang, Zhen; Zhang, Bowen; Chen, Kequan; Ouyang, Pingkai

    2016-02-01

    In this study, Actinobacillus succinogenes NJ113 microbial electrolysis cells (MECs) were used to enhance the reducing power responsible for succinic acid production from corncob hydrolysate. During corncob hydrolysate fermentation, electric MECs resulted in a 1.31-fold increase in succinic acid production and a 1.33-fold increase in the reducing power compared with those in non-electric MECs. When the hydrolysate was detoxified by combining Ca(OH)2, NaOH, and activated carbon, succinic acid production increased from 3.47 to 6.95 g/l. Using a constant potential of -1.8 V further increased succinic acid production to 7.18 g/l. A total of 18.09 g/l of succinic acid and a yield of 0.60 g/g total sugar were obtained after a 60-h fermentation when NaOH was used as a pH regulator. The improved succinic acid yield from corncob hydrolysate fermentation using A. succinogenes NJ113 in electric MECs demonstrates the great potential of using biomass as a feedstock to cost-effectively produce succinate.

  20. Enhanced antibacterial effect of antibiotics in combination with silver nanoparticles against animal pathogens.

    Science.gov (United States)

    Smekalova, Monika; Aragon, Virginia; Panacek, Ales; Prucek, Robert; Zboril, Radek; Kvitek, Libor

    2016-03-01

    Antibiotic resistant bacteria are a serious health risk in both human and veterinary medicine. Several studies have shown that silver nanoparticles (AgNPs) exert a high level of antibacterial activity against antibiotic resistant strains in humans. The aim of this study was to evaluate the antibacterial effects of a combined therapy of AgNPs and antibiotics against veterinary bacteria that show resistance to antibiotics. A microdilution checkerboard method was used to determine the minimal inhibitory concentrations of both types of antimicrobials, alone and in combination. The fractional inhibitory concentration index was calculated and used to classify observed collective antibacterial activity as synergistic, additive (only the sum of separate effects of drugs), indifferent (no effect) or antagonistic. From the 40 performed tests, seven were synergistic, 17 additive and 16 indifferent. None of the tested combinations showed an antagonistic effect. The majority of synergistic effects were observed for combinations of AgNPs given together with gentamicin, but the highest enhancement of antibacterial activity was found with combined therapy together with penicillin G against Actinobacillus pleuropneumoniae. A. pleuropneumoniae and Pasteurella multocida originally resistant to amoxycillin, gentamicin and colistin were sensitive to these antibiotics when combined with AgNPs. The study shows that AgNPs have potential as adjuvants for the treatment of animal bacterial diseases.

  1. Eikenella corrodens: Patogénesis y aspectos clínicos

    Directory of Open Access Journals (Sweden)

    Rubén Darío Jaramillo

    2006-09-01

    Full Text Available El ambiente microbiológico oral es único y tiene una dinámica compleja. Se calcula que cerca de 500 especies de bacterias habitan la cavidad oral humana, y alrededor de 22 géneros son los predominantes. Las bacterias que se aíslan con más frecuencia de los sitios infectados de la cavidad oral, y que son también patógenos potenciales, forma un grupo pequeño de microorganismos gramnegativos, entre los que se incluyen los siguientes: Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter spp., Capnocytophoga spp., Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia y el grupo-Streptococcus milleri. E. corrodens se reconoce como un microbio patógeno oportunista en la cavidad oral; también, puede causar infecciones extra orales, como un agente infeccioso solo o en combinaciones con diversas bacterias. La presente revisión, con un énfasis en E. corrodens, mostrará varios aspectos de sus características microbiológicas y bioquímicas, también se sistematizan y discuten el conocimiento actual sobre los mecanismos de patogénesis, como los lipopolisacáridos, proteínas externas de membrana, complejo de adesina, pili y otros; infecciones oral y extra oral; factores predisponentes; endocarditis; osteomielitis, infecciones intra-abdominales; diagnóstico y agentes antimicrobianos.

  2. Eikenella corrodens: Patogénesis y aspectos clínicos.

    Directory of Open Access Journals (Sweden)

    Rubén Darío Jaramillo

    2009-11-01

    Full Text Available El ambiente microbiológico oral es único y tiene una dinámica compleja. Se calcula que cerca de 500 especies de bacterias habitan la cavidad oral humana, y alrededor de 22 géneros son los predominantes. Las bacterias que se aíslan con más frecuencia de los sitios infectados de la cavidad oral, y que son también patógenos potenciales, forma un grupo pequeño de microorganismos gramnegativos, entre los que se incluyen los siguientes: Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter spp., Capnocytophoga spp., Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia y el grupo-Streptococcus milleri. E. corrodens se reconoce como un microbio patógeno oportunista en la cavidad oral; también, puede causar infecciones extra orales, como un agente infeccioso solo o en combinaciones con diversas bacterias. La presente revisión, con un énfasis en E. corrodens, mostrará varios aspectos de sus características microbiológicas y bioquímicas, también se sistematizan y discuten el conocimiento actual sobre los mecanismos de patogénesis, como los lipopolisacáridos, proteínas externas de membrana, complejo de adesina, pili y otros; infecciones oral y extra oral; factores predisponentes; endocarditis; osteomielitis, infecciones intra-abdominales; diagnóstico y agentes antimicrobianos.

  3. Isolation and identification of pathogenic bacteria from genital tract of the Arabian mares affected with genital tract infection and antimicrobial sensitivity

    Directory of Open Access Journals (Sweden)

    H. F. AL-Abidy

    2010-01-01

    Full Text Available This study was conducted for isolation and identification of the pathogenic bacteria presented in the genital tract infectionof the Arabian mares, and shows the anti microbial sensitivity. The study included 75 samples taken from infected maressuffering from genital tract infection diagnosed on the basis of case history and clinical signs which included bloody purulentdischarge ranched from yellow to green in colure, fetid oder with congested and oedematous vagina and from some abortioncases, and from mares suffered from tetanus disease symptoms during the period between October 2007 to April 2008 in studfarms breeding mares in Mosul. The samples were collected by swabs from the clitoris, clitorial fossa and the vagina. Isolationof bacteria was performed using aerobic and anaerobic culture techniques. Results of the present study showed a total ofisolation 75% from all samples taken with a high percentage isolation of Clostridium tetani (16.6%, followed by Archanobacterium pyogenes (10.6%, and Pseudomonas aeruginosa (8%, (6.7% for each Enterobacter aerogenes, Klebsiellapneumonia, Streptococcus dysagalactiae subsp equisimilis, and (5.3% for each bacteria Actinobacillus equilli, Streptococcuszooepidemicus, Staphylococcus aureus, then Proteus vulgaris (2.6%, and Escherichia coli (1.3%. The most bacterial isolateswere resistant to amoxicillin (100%, ampicillin (90.9 %, and erythromycin (65.9%, while the most isolates were sensitive tokanamycin (70.4%. It could be concluted that the most important bacteria causing genital tract infection of mares could beClostridium tetani and Archanobacterium pyogenes, Pseudomonas aeruginosa. The most bacterial isolates were resistant toamoxicillin, ampicillin and erythromycin.

  4. Uptake of photosensitizers by bacteria is influenced by the presence of cations

    Science.gov (United States)

    Kishen, A.; George, S.

    2007-05-01

    This investigation studies the influence of cations on photosensitizer uptake by Enterococcus faecalis (gram positive) and Actinobacillus actinomycetemcomitans (gram negative). Methods- The uptake of Methylene blue (MB) and Indocyanine Green (ICG), by bacteria were studied under the influence of divalent cations (CaCl II & MgCl II) and EDTA. Further, E. faecalis cells subjected to trypsinisation and calcium channel blocker (verapamil) were also analysed for MB and ICG uptake inorder to understand the mechanism of photosensitizer uptake. Results- Uptake of ICG was enhanced in the presence of divalent cations in E. faecalis and A. actinomycetemcomitans. Treating cells with EDTA had no significant effect on the photosensitizer uptake, although the highest concentration tested showed an enhancement of uptake. In contrast to ICG, MB showed a decreased uptake by bacterial cells on subjecting them to divalent cations and EDTA. Calcium channel blocker had no significant inhibitory effect on photosensitizers uptake. However, trypsin treatment resulted in significant reduction of ICG uptake. The result suggested that ICG uptake by bacteria is mediated through specific transporter protein while MB is associated with the outer surface structures of bacterial cells.

  5. [Susceptibility of potential periodontopathic bacteria to metronidazole, spiramycin and their combination].

    Science.gov (United States)

    Mouton, C; Dextraze, L; Mayrand, D

    1984-03-01

    A total of 65 bacterial strains originating mostly from subgingival plaque were tested for their susceptibilities to metronidazole, spiramycin, and their combination, ornidazole, erythromycin and tetracycline by means of an agar dilution technique. All agents were active against all anaerobic Gram-negative rods. Bacteroides gingivalis and Fusobacterium nucleatum showed marked susceptibility to metronidazole (MIC less than or equal to 0.06 microgram/ml) whereas 4-64 micrograms/ml were required to inhibit the capnophilic Actinobacillus actinomycetemcomitans and Capnocytophaga. Gram-positive facultatives were resistant to nitro-imidazoles but were inhibited at macrolide concentrations less than or equal to 0.5 microgram/ml. Except for F. nucleatum and Veillonella strains (2 less than or equal to MIC less than or equal to 128 micrograms/ml) macrolides were active against all other anaerobic bacteria tested. At concentrations less than or equal to 2 micrograms/ml the combination of spiramycin and metronidazole (2 : 1) was active against virtually all bacteria tested but our results failed to show a synergistic effect.

  6. PREVALENCE AND ANTIMICROBIAL RESISTANCE ASSESSMENT OF SUBCLINICAL MASTITIS IN MILK SAMPLES FROM SELECTED DAIRY FARMS

    Directory of Open Access Journals (Sweden)

    Murugaiyah Marimuthu

    2014-01-01

    Full Text Available This study was conducted in order to determine the prevalence and bacteriological assessment of subclinical mastitis and antimicrobial resistance of bacterial isolates from dairy cows in different farms around Selangor, Malaysia. A total of 120 milk samples from 3 different farms were randomly collected and tested for subclinical mastitis using California Mastitis Test (CMT, as well as for bacterial culture for isolation, identification and antimicrobial resistance. The most prevalent bacteria was Staphylococcus sp. (55%, followed by Bacillus sp., (21% and Corynebacterium sp., (7%, Yersinia sp. and Neisseria sp. both showed 5% prevalence, other species with prevalence below 5% are Acinetobacter sp., Actinobacillus sp., Vibrio sp., Pseudomonas sp., E.coli, Klebsiella sp. and Chromobacter sp. Selected Staphylococcus sp. showed a mean antimicrobial resistance of 73.3% to Ampicillin, 26.7% to Penicillin, Methicillin and Compound Sulphonamide each, 20% to Oxacillin, Amoxycillin and Cefuroxime, 13.3% to Polymyxin B, Erythromycin, Ceftriaxone and Azithromycin and 6.7% to Streptomycin, Clindamycin, Lincomycin and Tetracycline each. This study indicates the need for urgent and effective control measures to tackle the increase in prevalence of subclinical mastitis and their antimicrobial resistance in the study area.

  7. Comparing the efficacy of hyper-pure chlorine-dioxide with other oral antiseptics on oral pathogen microorganisms and biofilm in vitro.

    Science.gov (United States)

    Herczegh, Anna; Gyurkovics, Milán; Agababyan, Hayk; Ghidán, Agoston; Lohinai, Zsolt

    2013-09-01

    This study examines the antibacterial properties of sodium hypochlorite (NaOCl), chlorhexidine gluconate (CHX), Listerine®, and high purity chlorine dioxide (Solumium, ClO2) on selected common oral pathogen microorganisms and on dental biofilm in vitro. Antimicrobial activity of oral antiseptics was compared to the gold standard phenol. We investigated Streptococcus mutans, Lactobacillus acidophilus, Enterococcus faecalis, Veillonella alcalescens, Eikenella corrodens, Actinobacillus actinomycetemcomitans and Candida albicans as some important representatives of the oral pathogens. Furthermore, we collected dental plaque from the upper first molars of healthy young students. Massive biofilm was formed in vitro and its reduction was measured after treating it with mouthrinses: CHX, Listerine® or hyper pure ClO2. Their biofilm disrupting effect was measured after dissolving the crystal violet stain from biofilm by photometer. The results have showed that hyper pure ClO2 solution is more effective than other currently used disinfectants in case of aerobic bacteria and Candida yeast. In case of anaerobes its efficiency is similar to CHX solution. The biofilm dissolving effect of hyper pure ClO2 is significantly stronger compared to CHX and Listerine® after 5 min treatment. In conclusion, hyper pure ClO2 has a potent disinfectant efficacy on oral pathogenic microorganisms and a powerful biofilm dissolving effect compared to the current antiseptics, therefore high purity ClO2 may be a new promising preventive and therapeutic adjuvant in home oral care and in dental or oral surgery practice.

  8. Characterization and comparison of the bacterial microbiota in different gastrointestinal tract compartments in horses.

    Science.gov (United States)

    Costa, M C; Silva, G; Ramos, R V; Staempfli, H R; Arroyo, L G; Kim, P; Weese, J S

    2015-07-01

    The advance of new sequencing technologies has allowed more comprehensive characterization of complex microbial communities, including the ones inhabiting the intestinal tract. The presence of extreme environmental filters, such as low pH, digestive enzymes and anaerobic conditions along the tract, acts on the selection of unique bacteria in each compartment. The intestinal microbiota has an enormous impact on the maintenance of health. However, data about the bacteria present in the different intestinal compartments of horses are sparse. In this study, high throughput sequencing was used to characterize and compare bacterial profiles from different intestinal compartments of 11 horses scheduled for euthanasia for reasons other than gastrointestinal problems. Marked differences among compartments even at high taxonomic levels were found, with Firmicutes comprising the main bacterial phylum in all compartments. Lactobacillus spp. and Sarcina spp. predominated in the stomach and a marked increase of Streptococcus spp. occurred in the duodenum. Actinobacillus and Clostridium sensu stricto were the most abundant genera in the ileum and '5 genus incertae sedis', a genus from the Subdivision 5 class of the Verrucomicrobia, was the most abundant from the large colon through feces. There was a significant increase in diversity towards the distal gut with similar profiles observed from the cecum through feces at the class level. The bacterial population comprising the equine intestinal tract varies greatly among compartments and fecal samples may be useful as representative of changes occurring in the distal compartments.

  9. New consensus guidelines from the Clinical and Laboratory Standards Institute for antimicrobial susceptibility testing of infrequently isolated or fastidious bacteria.

    Science.gov (United States)

    Jorgensen, James H; Hindler, Janet F

    2007-01-15

    The Clinical and Laboratory Standards Institute (CLSI) recently published a new laboratory guideline for antimicrobial susceptibility testing of infrequently encountered or fastidious bacteria not covered in previous CLSI publications. The organisms include Aeromonas species, Bacillus species, and Vibrio species that may cause infections following environmental exposure. Fastidious organisms that may cause endocarditis or medical device infections include Abiotrophia and Granulicatella species; coryneform bacteria; Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella group gram-negative rods; and the instrinsically vancomycin-resistant gram-positive organisms Erysipelothrix, Lactobacillus, Leuconostoc, and Pediococcus species. Organisms not previously covered in depth in CLSI guidelines include Branhamella catarrhalis, Campylobacter jejuni, Campylobacter coli, Listeria species, and Pasteurella species. Clinically important drug resistance has been reported for each of these organisms. The guidelines provide recommendations for when it may be important to test these organisms, how standard methods may be easily adapted for testing, and appropriate interpretive criteria for results. Communication with infectious diseases clinicians prior to performing such testing is emphasized.

  10. Channel formation by RTX-toxins of pathogenic bacteria: Basis of their biological activity.

    Science.gov (United States)

    Benz, Roland

    2016-03-01

    The pore-forming cytolysins of the RTX-toxin (Repeats in ToXin) family are a relatively small fraction of a steadily increasing family of proteins that contain several functionally important glycine-rich and aspartate containing nonapeptide repeats. These cytolysins produced by a variety of Gram-negative bacteria form ion-permeable channels in erythrocytes and other eukaryotic cells. Hemolytic and cytolytic RTX-toxins represent pathogenicity factors of the toxin-producing bacteria and are very often important key factors in pathogenesis of the bacteria. Channel formation by RTX-toxins lead to the dissipation of ionic gradients and membrane potential across the cytoplasmic membrane of target cells, which results in cell death. Here we discuss channel formation and channel properties of some of the best known RTX-toxins, such as α-hemolysin (HlyA) of Escherichia coli and the uropathogenic EHEC strains, the adenylate cyclase toxin (ACT, CyaA) of Bordetella pertussis and the RTX-toxins (ApxI, ApxII and ApxIII) produced by different strains of Actinobacillus pleuropneumoniae. The channels formed by these RTX-toxins in lipid bilayers share some common properties such as cation selectivity and voltage-dependence. Furthermore the channels are transient and show frequent switching between different ion-conducting states. This article is part of a Special Issue entitled: Pore-Forming Toxins edited by Mauro Dalla Serra and Franco Gambale.

  11. Genetic organisation of the capsule transport gene region from Haemophilus paragallinarum

    Directory of Open Access Journals (Sweden)

    O. De Smidt

    2004-11-01

    Full Text Available The region involved in export of the capsule polysaccharides to the cell surface of Haemophilus paragallinarum was cloned and the genetic organisation determined. Degenerate primers designed from sequence alignment of the capsule transport genes of Haemophilus influenzae, Pasteurella multocida and Actinobacillus pleuropneumoniae were used to amplify a 2.6 kb fragment containing a segment of the H. paragallinarum capsule transport gene locus. This fragment was used as a digoxigenin labelled probe to isolate the complete H. paragallinarum capsule transport gene locus from genomic DNA. The sequence of the cloned DNA was determined and analysis revealed the presence of four genes, each showing high homology with known capsule transport genes. The four genes were designated hctA, B, C and D (for H. paragallinarum capsule transport genes and the predicted products of these genes likely encode an ATP-dependent export system responsible for transport of the capsule polysaccharides to the cell surface, possibly a member of a super family designated ABC (ATP-binding cassette transporters.

  12. Subgingival bacteria in a case of prepubertal periodontitis, before and one year after extractions of the affected primary teeth.

    Science.gov (United States)

    Ram, D; Bimstein, E

    1994-01-01

    The treatment of children with prepubertal periodontitis (PP), may be complicated by the extent of the lesions and the possibility of tetracycline stain of the developing permanent dentition. Therefore, with the purpose of preventing the infection of permanent teeth during the mixed dentition, it has been recommended that the treatment of children with PP, should include the early extraction of the primary teeth affected with alveolar bone loss (ABL). Still, there is little evidence which confirms that extraction of the affected primary teeth do in fact reduce the periodonto-pathogens load of the subgingival plaque. The present study reports values of colony forming units (CFU) of total anaerobic bacteria, Actinobacillus actynomicetemcomitans (Aa) and Porphyromonas gingivalis (Pg) from the subgingival plaque from a child with PP, collected immediately before and 1 year after extractions of the primary teeth affected with ABL. CFU of Aa and Pg developed only from the subgingival plaque collected before the extraction of the primary teeth affected with ABL. These findings suggest that in cases of PP, extraction of the affected primary teeth may reduce the possibility of infection of the periodontum of the permanent teeth during the mixed dentition period.

  13. Microbiological features of Papillon-Lefèvre syndrome periodontitis.

    Science.gov (United States)

    Velazco, C H; Coelho, C; Salazar, F; Contreras, A; Slots, J; Pacheco, J J

    1999-09-01

    Papillon-Lefevre syndrome patients exhibit hyperkeratosis palmo-plantaris and severe periodontitis. The syndrome is an autosomal recessive trait, but the mechanism of periodontal destruction is not known. This report presents the clinical and microbiological features of an 11-year old girl with Papillon-Lefèvre syndrome. Clinical examination included conventional periodontal measurements and radiographic analysis. In samples from 3 deep periodontal lesions, the occurrence of major suspected periodontopathic bacteria was determined by selective and non-selective culture and polymerase chain reaction (PCR) identification, and the presence of cytomegalovirus and Epstein-Barr type 1 virus by a nested-PCR detection method. 10 of 22 available teeth demonstrated severe periodontal breakdown. Major cultivable bacteria included Actinobacillus actinomycetemcomitans (3.4% of total isolates), Prevotella nigrescens (16.4%), Fusobacterium nucleatum (14.3%) and Peptostreptococcus micros (10.6%). A. actinomycetemcomitans, P. nigrescens, Porphyromonas gingivalis and Eikenella corrodens were identified by PCR analysis. The patient's non-affected parents and older brother revealed several periodontal pathogens but not A. actinomycetemcomitans. The viral examination demonstrated cytomegalovirus and Epstein-Barr type 1 virus in the subgingival sample of the Papillon-Lefèvre syndrome patient. The father and brother yielded subgingival cytomegalovirus but not Epstein-Barr type 1 virus. We hypothesize that human herpesviruses in concert with A. actinomycetemcomitans play important rôles in the development of Papillon-Lefèvre syndrome periodontitis.

  14. In vitro sensitivity of oral, gram-negative, facultative bacteria to the bactericidal activity of human neutrophil defensins.

    Science.gov (United States)

    Miyasaki, K T; Bodeau, A L; Ganz, T; Selsted, M E; Lehrer, R I

    1990-12-01

    Neutrophils play a major role in defending the periodontium against infection by oral, gram-negative, facultative bacteria, such as Actinobacillus actinomycetemcomitans, Eikenella corrodens, and Capnocytophaga spp. We examined the sensitivity of these bacteria to a mixture of low-molecular-weight peptides and highly purified individual defensin peptides (HNP-1, HNP-2, and HNP-3) isolated from human neutrophils. Whereas the Capnocytophaga spp. strains were killed significantly by the mixed human neutrophil peptides, the A. actinomycetemcomitans and E. corrodens strains were resistant. Killing was attributable to the defensins. The bactericidal activities of purified defensins HNP-1 and HNP-2 were equal, and both of these activities were greater than HNP-3 activity against strains of Capnocytophaga sputigena and Capnocytophaga gingivalis. The strain of Capnocytophaga ochracea was more sensitive to defensin-mediated bactericidal activity than either C. sputigena or C. gingivalis was. The three human defensins were equipotent in killing C. ochracea. C. ochracea was killed under aerobic and anaerobic conditions and over a broad pH range. Killing was most effective under hypotonic conditions but also occurred at physiologic salt concentrations. We concluded that Capnocytophaga spp. are sensitive to oxygen-independent killing by human defensins. Additional studies will be required to identify other components that may equip human neutrophils to kill A. actinomycetemcomitans, E. corrodens, and other oral gram-negative bacteria.

  15. Th1 and Th2 cytokine profile in patients with early onset periodontitis and their healthy siblings

    Directory of Open Access Journals (Sweden)

    Jiřina Bártová

    2000-01-01

    Full Text Available Early onset periodontitis (EOP is a chronic inflammatory periodontal disease with a strong genetic link affecting individuals aged 17 to 25. In the familial studies we tested the hypothesis about the role of Th1 and Th2 cytokines in the pathogenesis of EOP disease. The study involved 6 individuals with EOP disease and their 6 siblings with healthy periodontium. Actinobacillus actinomycetemcomitans (A. a., a bacterium typical for EOP, was detected in all people studied. Th1 and Th2 cytokine production was measured after in vitro stimulation. Peripheral blood mononuclear cells (PBMC were isolated and cultivated for 24 h and 7 days with PWM, A. a. or Escherichia coli. The levels of IL–4, IFN-gamma, IgA, IgG and IgM were measured by ELISA methods. After in vitro stimulation of PBMC, a significantly higher production of IL–4 and significantly lower production of IFN-gamma were found in the group of patients compared with their healthy siblings. The increased level of IL–4 in patients was in good agreement with an increased level of IgM after stimulation of lymphocytes with E. coli. These results support Seymour’s hypothesis according to which patients with progressive disease primarily activate Th2 lymphocytes while non-susceptible individuals activate Th1 lymphocytes.

  16. Expression of coding (mRNA) and non-coding (microRNA) RNA in lung tissue and blood isolated from pigs suffering from bacterial pleuropneumonia

    DEFF Research Database (Denmark)

    Skovgaard, Kerstin; Schou, Kirstine Klitgaard; Wendt, Karin Tarp

    2010-01-01

    the impact of microRNAs in the development and pathogenesis of lung infections. Expression of microRNA known to be induced by bacterial (i.e., LPS) ligands and thus supposed to play a role in the regulation of antimicrobial defence, were studied in lung tissue and in blood from pigs experimentally infected...... and to a lesser degree miR- 155 in lung tissue of the AP infected animals. MiR-233 was also found to be up regulated in blood based on both microarray and real-time PCR. Mir-233 has been found to be a negative regulator of neutrophil proliferation and activation, and might act to limit the potentially harmful...... with Actinobacillus pleuropneumoniae (AP). Expression differences of mRNA and microRNA were quantified at different time points (6h, 12h, 24h, 48h PI) using reverse transcription quantitative real-time PCR (Rotor-Gene and Fluidigm). Expression profiles of miRNA in blood of seven animals were further studied using mi...

  17. PnuC and the utilization of the nicotinamide riboside analog 3-aminopyridine in Haemophilus influenzae.

    Science.gov (United States)

    Sauer, Elizabeta; Merdanovic, Melisa; Mortimer, Anne Price; Bringmann, Gerhard; Reidl, Joachim

    2004-12-01

    The utilization pathway for the uptake of NAD and nicotinamide riboside was previously characterized for Haemophilus influenzae. We now report on the cellular location, topology, and substrate specificity of PnuC. pnuC of H. influenzae is only distantly related to pnuC of Escherichia coli and Salmonella enterica serovar Typhimurium. When E. coli PnuC was expressed in an H. influenzae pnuC mutant, it was able to take up only nicotinamide riboside and not nicotinamide mononucleotide. Therefore, we postulated that PnuC transporters in general possess specificity for nicotinamide riboside. Earlier studies showed that 3-aminopyridine derivatives (e.g., 3-aminopyridine adenine dinucleotide) are inhibitory for H. influenzae growth. By testing characterized strains with mutations in the NAD utilization pathway, we show that 3-aminopyridine riboside is inhibitory to H. influenzae and is taken up by the NAD-processing and nicotinamide riboside route. 3-Aminopyridine riboside is utilized effectively in a pnuC+ background. In addition, we demonstrate that 3-aminopyridine adenine dinucleotide resynthesis is produced by NadR. 3-Aminopyridine riboside-resistant H. influenzae isolates were characterized, and mutations in nadR could be detected. We also tested other species of the family Pasteurellaceae, Pasteurella multocida and Actinobacillus actinomycetemcomitans, and found that 3-aminopyridine riboside does not act as a growth inhibitor; hence, 3-aminopyridine riboside represents an anti-infective agent with a very narrow host range.

  18. Phylogeny of the genus Haemophilus as determined by comparison of partial infB sequences.

    Science.gov (United States)

    Hedegaard, J; Okkels, H; Bruun, B; Kilian, M; Mortensen, K K; Nørskov-Lauritsen, N

    2001-09-01

    A 453 bp fragment of infB, the gene encoding translation initiation factor 2, was sequenced and compared from 66 clinical isolates and type strains of Haemophilus species and related bacteria. Analysis of the partial infB sequences obtained suggested that the human isolates dependent on X and V factor, H. influenzae, H. haemolyticus, H. aegyptius and some cryptic genospecies of H. influenzae, were closely related to each other. H. parainfluenzae constituted a heterogeneous group within the boundaries of the genus, whereas H. aphrophilus/paraphrophilus and Actinobacillus actinomycetemcomitans were only remotely related to the type species of the genus Haemophilus H. parahaemolyticus and H. paraphrohaemolyticus took up an intermediary position and may not belong in the genus Haemophilus sensu stricto. Ambiguous results were obtained with seven isolates tentatively identified as H. segnis, which fell into two discrete clusters. The delineation of 'Haemophilus sensu stricto' as suggested by infB analysis supports previous results obtained by DNA hybridization, in contrast to the delineation inferred from 16S rRNA sequence comparison.

  19. Antimicrobial susceptibility monitoring of respiratory tract pathogens isolated from diseased cattle and pigs across Europe: the VetPath study.

    Science.gov (United States)

    de Jong, Anno; Thomas, Valérie; Simjee, Shabbir; Moyaert, Hilde; El Garch, Farid; Maher, Kirsty; Morrissey, Ian; Butty, Pascal; Klein, Ulrich; Marion, Hervé; Rigaut, Delphine; Vallé, Michel

    2014-08-06

    VetPath is an ongoing pan-European antibiotic susceptibility monitoring programme collecting pathogens from diseased antimicrobial non-treated cattle, pigs and poultry. In the current study, 1001 isolates from cattle and pig respiratory tract infections were tested for their antimicrobial susceptibilities. Non-replicate lung samples or nasopharyngeal/nasal swabs were collected from animals with acute clinical signs in 11 countries during 2002-2006. Pasteurella multocida and Mannheimia haemolytica from cattle and P. multocida, Actinobacillus pleuropneumoniae and Streptococcus suis from pigs were isolated by standard methods. S. suis was also isolated from meningitis cases. MICs of 16 antibiotics were assessed centrally by broth microdilution following CLSI recommendations. Results were interpreted using CLSI breakpoints where available. P. multocida (231) and M. haemolytica (138) isolates were all susceptible to amoxicillin/clavulanic acid, ceftiofur, enrofloxacin and trimethoprim/sulfamethoxazole. Resistance to florfenicol and spectinomycin was 0.4% and 3.5% in P. multocida, respectively, and absent in M. haemolytica isolates. Tetracycline resistance was 5.7% and 14.6% for P. multocida and M. haemolytica. In pigs, 230 P. multocida, 220 A. pleuropneumoniae and 182 S. suis isolates were recovered. Resistance to amoxicillin/clavulanic acid, ceftiofur, enrofloxacin, florfenicol, tiamulin and tilmicosin was absent or cattle and pigs. Since for approximately half of the antibiotics in this panel no CLSI-defined breakpoints were available, setting of the missing veterinary breakpoints is important.

  20. Erythema Multiforme Associated with Respiratory Disease in a Commercial Breeding Pig Herd.

    Science.gov (United States)

    Papatsiros, Vasileios G; Athanasiou, Labrini V; Psalla, Dimitra; Petridou, Evanthia; Maragkakis, Giorgos G; Papatsas, Ioannis; Arsenakis, Ioannis; Maes, Dominiek

    2015-10-01

    This study describes an erythema multiforme (EM) in breeding sows, after their mixing in the group housing system. Sows at 30-35 days of gestation showed red and raised skin areas, depression, anorexia, fever, respiratory problems, and increased return to estrus. Blood and nasal samples from diseased sows were examined by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay for respiratory pathogens. Hematological and biochemical analyses were performed on the blood samples. From diseased sows, vaginal swabs for microbiological examinations and samples at slaughterhouse for gross and microscopic examinations were collected. Samples from the complete gestation and lactation feed were examined for mycotoxins. All sampled sows were seropositive for Actinobacillus pleuropneumoniae (App) and porcine reproductive and respiratory syndrome virus (PRRSV). No viremia for PRRSV and porcine circovirus type 2 were detected. All nasal samples were positive for Streptococcus suis, one for Swine Influenza Virus and one for App, Hemophilus parasuis, and S. suis. In all vaginal swabs, Escherichia coli and Streptococcus spp. were detected. Diseased sows had moderate leukocytosis, mild anemia, and thrombocytopenia. No mycotoxins were detected in feed. Histopathological examination revealed increased vascularization of the superficial and middle dermis. EM was likely due to illness caused by viral and bacterial infections. This study suggests that stress caused by the sows' mixing might have triggered the problem.

  1. Integration host factor is required for replication of pYGK-derived plasmids in Aggregatibacter actinomycetemcomitans.

    Science.gov (United States)

    Torres-Escobar, Ascención; Juárez-Rodríguez, María D; Demuth, Donald R

    2014-08-01

    In this study, we show that integration host factor protein (IHF) is required for replication of pYGK plasmids in Aggregatibacter actinomycetemcomitans. YGK plasmids were not replicated in A. actinomycetemcomitans strains lacking either the α- or β- subunit of IHF. However, the deletion mutants were complemented, and plasmid replication was restored when the promoter region and gene for either ihfA or ihfB was cloned into pYGK. We also identified two motifs that resemble the consensus IHF-binding site in a 813-bp fragment containing the pYGK origin of replication. Using electrophoretic mobility shift assays, purified IHFα-IHFβ protein complex was shown to bind to probes containing either of these motifs. To our knowledge, this is the first report showing that plasmid replication is IHF-dependent in the family Pasteurellaceae. In addition, using site-direct mutagenesis, the XbaI and KpnI restriction sites in the suicide vector pJT1 were modified to generate plasmid pJT10. The introduction of these new unique sites in pJT10 facilitates the transfer of transcriptional or translational lacZ fusion constructs for the generation of single-copy chromosomal insertion of the reporter construct. Plasmid pJT10 and its derivatives will be useful for genetic studies in Aggregatibacter (Actinobacillus) and probably other genera of Pasteurellaceae, including Haemophilus, Pasteurella, and Mannheimia.

  2. Bacterial endocarditis prophylaxis.

    Science.gov (United States)

    Blanco-Carrión, Andrés

    2004-01-01

    Bacterial endocarditis (BE) is a disease resulting from the association of morphological alterations of the heart and bacteraemia originating from different sources that at times can be indiscernible (infectious endocarditis). It is classified on the basis of the morphological alteration involved, depending on the clinical manifestations and course of illness, which varies according to the causative microorganism and host conditions (for example, it is characteristic in I.V. drug users). The most common microorganisms involved are: Streptococcus viridans (55%), Staphylococcus aureus (30%), Enterococcus (6%) and HACEK bacteria (corresponding to the initials: Haemophilus, Actinobacillus, Cardiobacterium, Eikenella and Kingella), although on occasions it can also be caused by fungi. The oral microbiological flora plays a very important role in the aetiopathogenesis of BE, given that the condition may be of oral or dental origin. This paper will deal with the prevention of said bacteraemia. Prophylaxis will be undertaken using amoxicillin or clindamycin according to action protocols, with special emphasis placed on oral hygiene in patients with structural defects of the heart.

  3. Rapid adaptation drives invasion of airway donor microbiota by Pseudomonas after lung transplantation.

    Science.gov (United States)

    Beaume, M; Köhler, T; Greub, G; Manuel, O; Aubert, J-D; Baerlocher, L; Farinelli, L; Buckling, A; van Delden, C

    2017-01-17

    In cystic fibrosis (CF) patients, chronic airway infection by Pseudomonas leads to progressive lung destruction ultimately requiring lung transplantation (LT). Following LT, CF-adapted Pseudomonas strains, potentially originating from the sinuses, may seed the allograft leading to infections and reduced allograft survival. We investigated whether CF-adapted Pseudomonas populations invade the donor microbiota and adapt to the non-CF allograft. We collected sequential Pseudomonas isolates and airway samples from a CF-lung transplant recipient during two years, and followed the dynamics of the microbiota and Pseudomonas populations. We show that Pseudomonas invaded the host microbiota within three days post-LT, in association with a reduction in richness and diversity. A dominant mucoid and hypermutator mutL lineage was replaced after 11 days by non-mucoid strains. Despite antibiotic therapy, Pseudomonas dominated the allograft microbiota until day 95. We observed positive selection of pre-LT variants and the appearance of novel mutations. Phenotypic adaptation resulted in increased biofilm formation and swimming motility capacities. Pseudomonas was replaced after 95 days by a microbiota dominated by Actinobacillus. In conclusion, mucoid Pseudomonas adapted to the CF-lung remained able to invade the allograft. Selection of both pre-existing non-mucoid subpopulations and of novel phenotypic traits suggests rapid adaptation of Pseudomonas to the non-CF allograft.

  4. High Sensitivity Crosslink Detection Coupled With Integrative Structure Modeling in the Mass Spec Studio.

    Science.gov (United States)

    Sarpe, Vladimir; Rafiei, Atefeh; Hepburn, Morgan; Ostan, Nicholas; Schryvers, Anthony B; Schriemer, David C

    2016-09-01

    The Mass Spec Studio package was designed to support the extraction of hydrogen-deuterium exchange and covalent labeling data for a range of mass spectrometry (MS)-based workflows, to integrate with restraint-driven protein modeling activities. In this report, we present an extension of the underlying Studio framework and provide a plug-in for crosslink (XL) detection. To accommodate flexibility in XL methods and applications, while maintaining efficient data processing, the plug-in employs a peptide library reduction strategy via a presearch of the tandem-MS data. We demonstrate that prescoring linear unmodified peptide tags using a probabilistic approach substantially reduces search space by requiring both crosslinked peptides to generate sparse data attributable to their linear forms. The method demonstrates highly sensitive crosslink peptide identification with a low false positive rate. Integration with a Haddock plug-in provides a resource that can combine multiple sources of data for protein modeling activities. We generated a structural model of porcine transferrin bound to TbpB, a membrane-bound receptor essential for iron acquisition in Actinobacillus pleuropneumoniae Using mutational data and crosslinking restraints, we confirm the mechanism by which TbpB recognizes the iron-loaded form of transferrin, and note the requirement for disparate sources of restraint data for accurate model construction. The software plugin is freely available at www.msstudio.ca.

  5. Enzymatic degradation of collagen-guided tissue regeneration membranes by periodontal bacteria.

    Science.gov (United States)

    Sela, Michael N; Kohavi, David; Krausz, Emanuela; Steinberg, Doron; Rosen, Graciela

    2003-06-01

    Bacterial infection in the vicinity of guided tissue regeneration barrier membranes was shown to have a negative effect on the clinical outcomes of this increasingly used technique. Several oral and specifically periodontal bacteria were shown to adhere to such membranes in vivo and in vitro with a higher affinity to membranes constructed from collagen. The present study examined the role of periodontal bacteria and their enzymes in the degradation of commercially used collagen membranes. Degradation of two collagen membranes [Biomend (Calcitek, Colla-Tec Inc., Plainsboro, NJ) and Bio-Gide (Geistlich Biomaterials, Wolhousen, Switzerland)] labeled by fluorescein isothiocyanate was examined by measuring soluble fluorescence. Porphyromonas gingivalis, Treponema denticola and Actinobacillus actinomycetemcomitans and their enzymes were evaluated. Collagenase from Clostridium hystolyticum was used as a positive control. While whole cells of P. gingivalis were able to degrade both types of membranes, T. denticola could degrade Bio-Gide membranes only and A. actinomycetemcomitans whole cells could degrade none of the membranes. Fractionation of P. gingivalis cells revealed that cell membrane associated proteases were responsible for the degradation of the two collagen membranes. In T. denticola, the purified major phenylalanine protease was found to be responsible for the degradation of Bio-Gide membranes. These results suggest that proteolytic bacterial enzymes may take part in the degradation of collagen barrier membranes used for guided tissue regeneration.

  6. Disease severity associated with presence in subgingival plaque of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Tannerella forsythia, singly or in combination, as detected by nested multiplex PCR.

    Science.gov (United States)

    Ready, D; D'Aiuto, F; Spratt, D A; Suvan, J; Tonetti, M S; Wilson, M

    2008-10-01

    This study used a nested multiplex PCR method to detect three periodontal pathogens in subgingival plaque collected before treatment and at 2 and 6 months posttreatment from 107 patients with severe, generalized periodontitis. The proportions of the patients who harbored these bacteria before periodontal treatment were as follows: Tannerella forsythia, 81%; Porphyromonas gingivalis, 78%; and Aggregatibacter (formerly Actinobacillus) actinomycetemcomitans, 47%. At 2 months posttreatment there was a significant reduction in the numbers of patients harboring P. gingivalis (46%; P < 0.001) or T. forsythia (63%; P = 0.043) but not A. actinomycetemcomitans (50%) compared to pretreatment data. At 6 months posttreatment, significantly fewer patients harbored P. gingivalis (43%; P < 0.001); A. actinomycetemcomitans, (31%; P = 0.025), or T. forsythia (63%; P = 0.030). Interestingly, at baseline and at 2 months posttherapy, subjects who harbored only a single pathogen had a greater level of periodontal disease than subjects who harbored two, or all three, of these periodontal pathogens. These data suggest that a reduction in the number of species present may be associated with an increase in the severity of periodontal diseases.

  7. 以白酒酒糟为原料发酵产丁二酸%Study on fermentative production of succinic acid from spirit-based distillers' grains

    Institute of Scientific and Technical Information of China (English)

    周小兵; 郑璞

    2013-01-01

    Spirit-based ditillers' grains ( SDG) treated with enzyme hydrolysis was used as a stuff for fermentative production of succinic acid by Actinobacillus succinogenes.When the hydrolysis reaction was solely catalyzed by cellulase or amylase under its optimum temperature and pH, the corresponding hydrolysis rates of cellulose in SDG and starch in SDG were 44.04% and 92.26% , respectively.While hydrolysis using cellulase and amylase step by step ( HSS) , the later hydrolysis reaction could be restrained.Otherwise, the yield of reducing sugar in hydrolysis u-sing cellulase and amylase simultaneously was about 150 mg/g SDG, the same as that of HSS.Using hydrolysis and fermentation separated process, 58.4 g/L reducing sugar was obtained, which resulted in 28.8 g/L succinic acid with a yield of 72 mg/g SDG.However, using the procedure that firstly SDG was hydrolyzed with cellulase and then starch in SDG was simultaneously saccharified and fermented, 32 g/L succinic acid was achieved with a yield of 133 mg/g SDG.This study provided a new economical stuff for fermentative production of succinic acid in the future.%以白酒酒糟为原料,经酶法糖化,由Actinobacillus succinogenes发酵生产丁二酸.纤维素酶或糖化酶分别水解白酒糟,在酶反应的最适温度和pH条件下,酒糟中的纤维素和淀粉的水解率分别为44.04%和92.26%,相应还原糖对酒糟的得率分别为110 mg/g和126 mg/g酒糟;但2种酶以分步或同步方式水解白酒糟时,酶水解反应受到产物抑制作用,总还原糖得率仅约为150 mg/g酒糟.采用分步糖化发酵工艺,400g/L白酒糟经两种酶水解后,得到还原糖58.4 g/L,该水解液发酵产丁二酸28.8g/L,丁二酸产率72 mg/g酒糟;而采用先用纤维素酶水解白酒糟,再用糖化酶和A.succinogenes同步糖化发酵的工艺,240 g/L白酒糟产丁二酸浓度为32 g/L,产率133 mg/g酒糟.以白酒酒糟为原料发酵生产丁二酸,利用了废弃物,无需外源添加氮源,

  8. Detection of five periodontal pathogens in coronary atherosclerotic plaque%牙周致病菌在动脉粥样硬化斑块中的检测

    Institute of Scientific and Technical Information of China (English)

    侯秀丽; 梁平; 张源明; 木拉提; 艾斯卡尔; 艾克拜尔; 闰跃斌

    2011-01-01

    目的 检测冠状动脉粥样硬化斑块中的5种特异性牙周致病菌.方法 收集101例行冠状动脉搭桥手术患者的动脉粥样硬化斑块标本,采用Chelex-100法提取冠状动脉粥样硬化斑块中的DNA,并通过PCR分别检测动脉粥样硬化斑块中的牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)、具核梭杆菌(Fusobacterium nucleatum,Fn)、中间普氏菌(Prevotella intermedia,Pi)、福赛斯坦纳菌(Tannerella forsythensis,Tf)5种牙周特异性致病菌.结果 101例患者动脉粥样硬化斑块中牙龈卟啉单胞菌(Pg)的检出率为31%,福赛斯坦纳菌(Tf)为42%,中间普氏菌(Pi)为26%,具核梭杆菌(Fn)为21%,伴放线放线杆菌(Aa)为23%.这几种细菌的PCR产物通过测序,结果与GenBank数据库中的序列进行比对同源性达99%~100%.结论 牙周致病菌在冠心病的发生、发展中可能发挥着一定作用,考虑慢性牙周炎与冠心病之间有一定的相关性.%Objective To detect five periodontal pathogenic bacteria in coronary atherosclerotic plaques.Methods Atherosclerotic plaque specimens were obtained from 101 patients who scheduled for coronary artery bypass graft surgery.The bacteria DNA was obtained from coronary atherosclerotic plaques using the chelex-100 method.The extracted DNA were examined by PCR.Results Within the 101 pieces of atherosclerotic plaque samples Porphyromonas gingivalis( Pg,31% ),Tannerella forsythensis(Tf,42% ),Prevotella intermedia( Pi,26% ),Fusobacterium nucleatum( Fn,21% ),Actinobacillus actinomycetemcomitans( Aa,23% ).PCR products were sequenced and were compared with GenBank sequences,the homology was 99%-100%.Conclusion Periodontitis might affect the development of atherosclerosis and there is a correlation between coronary heart disease and chronic periodontitis.

  9. Estudio microbiológico de la periimplantitis: Presentación de 9 casos clínicos Microbiological study of the peri-implantitis: presentation of 9 clinical case

    Directory of Open Access Journals (Sweden)

    M. Quinteros Borgarello

    2000-12-01

    Full Text Available Objetivos: La periimplantitis es un proceso inflamatorio de los tejidos que rodean a un implante sometido a carga que produce la pérdida de soporte óseo. En este trabajo se muestran los resultados del estudio microbiológico y del antibiograma realizado a partir del exudado y/o del tejido periimplantario de 16 implantes funcionales con periimplantitis de 9 pacientes que fueron tratados en nuestro servicio. Material y método: Se estudiaron 16 periimplantitis observadas en 9 pacientes; de 14 de ellas se obtuvieron muestras con puntas de papel estériles que se introdujeron en la bolsa periimplantaria. En 2 implantes las muestras procedian de tejido de granulación periimplantario. Inmediatamente después de ser obtenidas, se colocaban en placas de agar-chocolate con brain heart infusion para proceder a su cultivo. Resultados: Los especímenes bacterianos aislados fueron: Stomatococcus, Prevotella oralis, Peptostreptococcus y Fusobacterium nucleatum. En 9 muestras fue imposible aislar una bacteria predominante debido a la complejidad de la flora. Respecto al resto de muestras, la bacteria predominante fue Stomatococcus en 3, Prevotella oralis en 1,Peptostreptococcus en 1 y Fusobacterium nucleatum en 2. Los antibiogramas mostraron una mayor sensibilidad a la asociación de la amoxicilina con el ácido clavulánico, comparada con la amoxicilina, el metronidazol o una combinación de estos dos últimos antimicrobianos. Conclusiones: En este trabajo las bacterias asociadas más frecuentemente a la periimplantitis fueron: Stomatococcus, Prevotella oralis, Peptostreptococcus y Fusobacterium nucleatum. En ningún caso se aislaron cepas de Actinobacillus actinomycetemcomitans. Los posibles factores de riesgo asociados a periimplantitis en nuestra serie fueron los implantes recubiertos con hidroxiapatita, implantes de 3,25mm de diámetro y la localización más distal del implante en las prótesis que rehabilitaban extremos libres edéntulos superiores

  10. Identification of the Antibacterial Activity ofMorus rubra Leaves by a High- Throughput Screening Method%以高通量方法筛选红桑叶的抗菌活性并进行活性组分的分离研究

    Institute of Scientific and Technical Information of China (English)

    周英; 黄赤夫

    2007-01-01

    目的 通过一种简单快速的高通量方法筛选具有抗菌活性的植物提取物,并通过此种方法从有活性的粗提物中纯化活性组分.方法 首先建立植物提取物库和抗菌的高通量筛选方法.然后针对转糖链球菌Streptococcus mutans和伴放线放线杆菌Actinobacillus actinomycetemcomitans进行高通量筛选.结果 通过筛选发现红桑叶具有很强的抑菌活性.为了进一步研究其活性,将其粗提物通过HPLC进行分离得到71个组分,再通过同样的高通量筛选方法进行抗菌活性测定,对有活性的组分通过血琼脂法进行验证.实验数据表明红桑叶的一个组分#31对两种细菌有几乎完全的抑制(抑制率>99%),而另外两个组分对伴放线放线杆菌Actinobacillus actinomycetemcomitans有部分抑制.结论 所建立的高通量筛选方法简便快捷,筛选结果稳定可靠,可用于大规模的植物提取物的抗菌活性筛选.红桑叶具有很强的抗菌活性,且可分离得到活性组分,红桑叶具有进一步开发为抗菌药物和保健品的潜力.%Objective This research intended to test the antibacterial activity of a plant extract library using a simple and fast colorimetric detection method and identify those plant extracts with bioactivity, and to further purify those bioactive fractions of the selected plant extracts. Methods We first created a plant extract library and developed an antimicrobial High Throughput Screening (HTS) method. Then, the plant extracts were tested for antibacterial activity against two species of bacteria, S. mutans and A. actinomycestemcomitans. Results The Morus rubra leaf extract was selected because it had the strongest biological activity against the oral bacteria tested. To further study the Morus rubra leaves, the extract was separated into small fractions via high -performance liquid chromatography (HPLC). The fractions were tested against bacteria S. mutans and A. actinomycestemcomitans using HTS

  11. Genomic characterization of Haemophilus parasuis SH0165, a highly virulent strain of serovar 5 prevalent in China.

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    Zhuofei Xu

    Full Text Available Haemophilus parasuis can be either a commensal bacterium of the porcine respiratory tract or an opportunistic pathogen causing Glässer's disease, a severe systemic disease that has led to significant economical losses in the pig industry worldwide. We determined the complete genomic sequence of H. parasuis SH0165, a highly virulent strain of serovar 5, which was isolated from a hog pen in North China. The single circular chromosome was 2,269,156 base pairs in length and contained 2,031 protein-coding genes. Together with the full spectrum of genes detected by the analysis of metabolic pathways, we confirmed that H. parasuis generates ATP via both fermentation and respiration, and possesses an intact TCA cycle for anabolism. In addition to possessing the complete pathway essential for the biosynthesis of heme, this pathogen was also found to be well-equipped with different iron acquisition systems, such as the TonB system and ABC-type transport complexes, to overcome iron limitation during infection and persistence. We identified a number of genes encoding potential virulence factors, such as type IV fimbriae and surface polysaccharides. Analysis of the genome confirmed that H. parasuis is naturally competent, as genes related to DNA uptake are present. A nine-mer DNA uptake signal sequence (ACAAGCGGT, identical to that found in Actinobacillus pleuropneumoniae and Mannheimia haemolytica, followed by similar downstream motifs, was identified in the SH0165 genome. Genomic and phylogenetic comparisons with other Pasteurellaceae species further indicated that H. parasuis was closely related to another swine pathogenic bacteria A. pleuropneumoniae. The comprehensive genetic analysis presented here provides a foundation for future research on the metabolism, natural competence and virulence of H. parasuis.

  12. APPROUSH TO SPECIFIC DIAGNOSTICS OF CAUSATIVE AGENTS OF INFECTIOUS ENDOCARDIDIS

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    Katsapov D.V.

    2015-05-01

    Full Text Available Introduction. Increased level of morbidity of infective endocarditis (IE connected with new risk factors: intravenous drug use, cardiosurgical interventions, hemodialysis brought new clinical forms of the disease. As it shown in a literature main pathogenetic factors of IE are bacteraemia, trauma of endocardium and invasive medical procedures. Very typical pathogens are streptococci and staphylococci. Most typically mitral and aortal valves are affected with spreading of vegetations on surrounding media. Discussion. IE is polyetiologic disease caused by more than 128 microorganisms, and still a challenge for medical professionals. Detection a causative agent is critical for proper specific treatment. In different sources data on percentage of proven cases very according to country and different medical centres reflecting different local epidemiology of IE, diagnostic criteria and protocols. Culture negative infectious endocarditis (CNIE is considered in case of obtaining of three negative results of cultivation of samples on a standard blood agar during 7 days and subculturing. CNIE incidence very form 2% to 33% according to different researches and higher in cases of community acquired infection and reseeding antibacterial treatment. Some of cases of CNIE caused by gram - negative fastidious microorganisms - Haemophilus parainfluenzae, Actinobacillus, Actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens, Kingella kingae, with united in HACEK group according to their properties to colonize oropharynx and requirement in special conditions and duration of incubation. Detection of some intracellular bacteria, such as C. burnetti and Bartonella spp. require immunological methods of detection, histological methods and of PCR. Conclusion. In case of diagnostics of patients with CNIE it is necessary to use a combination of prolonged subculturing of serum, emboli and histologic material on blood agar with microscopy by Warthin

  13. The use of ozonated water and 0.2% chlorhexidine in the treatment of periodontitis patients: A clinical and microbiologic study

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    Kshitish Durga

    2010-01-01

    Full Text Available Background: The development of periodontal disease has been thought to be associated with several restricted members of the oral anaerobic species, such as black-pigmented Porphyromonas species and Actinobacillus actinomycetemcomitans (Aa, in the subgingival environment. Apart from bacteria, certain viruses and fungi that are associated with periodontal disease are also present in the subgingival plaque . Materials and Methods: A randomized, double-blind, crossover split-mouth design was performed. A total of 16 patients suffering from generalized chronic periodontitis were selected for the study. The study period of 18 days was divided into two time-intervals, i.e. baseline (0 days to 7 th day, with a washout period of 4 days followed by a second time interval of 7 days. The use of ozone and chlorhexidine gluconate (CHX irrigation was randomized. Both the patient and the clinician evaluating the clinical parameters were blinded regarding the type of irrigation used. Results: The interpretation of clinical and microbial data is from baseline to 7 th day. A higher percentage of plaque index (12%, gingival index (29% and bleeding index (26% reduction was observed using ozone irrigation as compared to chlorhexidine. The percentile reduction of Aa (25% using ozone was appreciable as compared to no change in Aa occurrence using chlorhexidine. By using O 3 and chlorhexidine, there was no antibacterial effect on Porphyromonas gingivalis (Pg and Tannerella forsythensis. The antifungal effect of ozone from baseline (37% to 7 th day (12.5% was pronounced during the study period, unlike CHX, which did not demonstrate any antifungal effect. Conclusion: Ozone may be considered as an alternative management strategy due to its powerful ability to inactivate microorganisms. Also, there is growing evidence that ozone can be employed as a useful therapeutic agent in both dentistry and medicine.

  14. Diversity of bacterial species in the nasal cavity of sheep in the highlands of Ethiopia and first report of Histophilus somni in the country.

    Science.gov (United States)

    Tesfaye, Biruk; Sisay Tessema, Tesfaye; Tefera, Genene

    2013-06-01

    A study was conducted to isolate bacterial species/pathogens from the nasal cavity of apparently healthy and pneumonic sheep. Nasal swabs were collected aseptically, transported in tryptose soya broth and incubated for 24 h. Then, each swab was streaked onto chocolate and blood agar for culture. Bacterial species were identified following standard bacteriological procedures. Accordingly, a total of 1,556 bacteria were isolated from 960 nasal swabs collected from three different highland areas of Ethiopia, namely Debre Berhan, Asella, and Gimba. In Debre Berhan, 140 Mannheimia haemolytica, 81 Histophilus somni, 57 Staphylococcus species, and 52 Bibersteinia trehalosi were isolated. While from Gimba M. haemolytica, Staphylococcus, Streptococcus, and H. somni were isolated at rates of 25.2, 15.9, 11.4, and 5.9 %, respectively, of the total 647 bacterial species. In Asella from 352 bacterial species isolated, 93 (26.4 %) were M. haemolytica, 48 (13.6 %) were Staphylococcus species, 26 (7.4 %) were B. trehalosi, and 17 (4.8 %) H. somni were recognized. Further identification and characterization using BIOLOG identification system Enterococcus avium and Sphingomonas sanguinis were identified at 100 % probability, while, H. somni and Actinobacillus lignerisii were suggested by the system. The study showed that a variety of bacterial species colonize the nasal cavity of the Ethiopian highland sheep with variable proportion between healthy and pneumonic ones. To our knowledge, this is the first report on isolation of H. somni, an important pathogen in cattle, from the respiratory tract of a ruminant species in the country.

  15. Host-Derived Sialic Acids Are an Important Nutrient Source Required for Optimal Bacterial Fitness In Vivo

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    Nathan D. McDonald

    2016-04-01

    Full Text Available A major challenge facing bacterial intestinal pathogens is competition for nutrient sources with the host microbiota. Vibrio cholerae is an intestinal pathogen that causes cholera, which affects millions each year; however, our knowledge of its nutritional requirements in the intestinal milieu is limited. In this study, we demonstrated that V. cholerae can grow efficiently on intestinal mucus and its component sialic acids and that a tripartite ATP-independent periplasmic SiaPQM strain, transporter-deficient mutant NC1777, was attenuated for colonization using a streptomycin-pretreated adult mouse model. In in vivo competition assays, NC1777 was significantly outcompeted for up to 3 days postinfection. NC1777 was also significantly outcompeted in in vitro competition assays in M9 minimal medium supplemented with intestinal mucus, indicating that sialic acid uptake is essential for fitness. Phylogenetic analyses demonstrated that the ability to utilize sialic acid was distributed among 452 bacterial species from eight phyla. The majority of species belonged to four phyla, Actinobacteria (members of Actinobacillus, Corynebacterium, Mycoplasma, and Streptomyces, Bacteroidetes (mainly Bacteroides, Capnocytophaga, and Prevotella, Firmicutes (members of Streptococcus, Staphylococcus, Clostridium, and Lactobacillus, and Proteobacteria (including Escherichia, Shigella, Salmonella, Citrobacter, Haemophilus, Klebsiella, Pasteurella, Photobacterium, Vibrio, and Yersinia species, mostly commensals and/or pathogens. Overall, our data demonstrate that the ability to take up host-derived sugars and sialic acid specifically allows V. cholerae a competitive advantage in intestinal colonization and that this is a trait that is sporadic in its occurrence and phylogenetic distribution and ancestral in some genera but horizontally acquired in others.

  16. Longitudinal study of respiratory infection patterns of breeding sows in five farrow-to-finish herds.

    Science.gov (United States)

    Fablet, C; Marois, C; Kuntz-Simon, G; Rose, N; Dorenlor, V; Eono, F; Eveno, E; Jolly, J P; Le Devendec, L; Tocqueville, V; Quéguiner, S; Gorin, S; Kobisch, M; Madec, F

    2011-01-27

    A longitudinal study was carried out in five French farrow-to-finish herds differently affected by respiratory diseases to describe the carrying and infection patterns of batches of sows to various respiratory pathogens during gestation and lactation. An entire batch of sows was followed during two successive reproduction cycles. Nasal, tonsillar and oro-pharyngeal swabs and blood samples were taken from each sow 9 and 4 weeks before farrowing and 1 and 4 weeks after farrowing. Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, Pasteurella multocida, Haemophilus parasuis and Streptococcus suis were detected from swab samples using PCR assays. Blood samples were tested for antibodies against M. hyopneumoniae, A. pleuropneumoniae serotypes 1-9-11 and 2, Porcine Circovirus type-2 (PCV-2) and Porcine Reproductive and Respiratory Syndrome virus (PRRSV) by ELISA tests. Antibodies against H(1)N(1), H(1)N(2) and H(3)N(2) Swine Influenza Viruses (SIV) of European lineages were tested by hemagglutination inhibition assay. The results indicated that S. suis is widespread among sows (67.1% of PCR-positive sows). A. pleuropneumoniae, P. multocida, and H. parasuis were detected by PCR in 30.9%, 24.6% and 23.4% of the sows, respectively. Antibodies against M. hyopneumoniae were recovered from more than 55% of the sows in all herds whereas the micro-organism was detected in 2.4% of the sows. Although PCV-2 and SIV infections were highly prevalent, the PRRSV infection patterns ranged from no infection in farms mildly affected by respiratory diseases to active circulation in more severely affected herds. The sow population thus constitutes a reservoir for a continuous circulation of respiratory pathogens and needs to be properly considered in control strategies.

  17. Serum IgG antibody levels to periodontal microbiota are associated with incident Alzheimer disease.

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    James M Noble

    Full Text Available Periodontitis and Alzheimer disease (AD are associated with systemic inflammation. This research studied serum IgG to periodontal microbiota as possible predictors of incident AD.Using a case-cohort study design, 219 subjects (110 incident AD cases and 109 controls without incident cognitive impairment at last follow-up, matched on race-ethnicity, were drawn from the Washington Heights-Inwood Columbia Aging Project (WHICAP, a cohort of longitudinally followed northern Manhattan residents aged >65 years. Mean follow-up was five years (SD 2.6. In baseline sera, serum IgG levels were determined for bacteria known to be positively or negatively associated with periodontitis (Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans Y4, Treponema denticola, Campylobacter rectus, Eubacterium nodatum, and Actinomyces naeslundii genospecies-2. In all analyses, we used antibody threshold levels shown to correlate with presence of moderate-severe periodontitis.Mean age was 72 years (SD 6.9 for controls, and 79 years (SD 4.6 for cases (p640 ng/ml, present in 10% of subjects was associated with increased risk of AD (HR = 2.0, 95%CI: 1.1-3.8. This association was stronger after adjusting for other significant titers (HR = 3.1, 95%CI: 1.5-6.4. In this model, high anti-E. nodatum IgG (>1755 ng/ml; 19% of subjects was associated with lower risk of AD (HR = 0.5, 95%CI: 0.2-0.9.Serum IgG levels to common periodontal microbiota are associated with risk for developing incident AD.

  18. Targeted profiling of oral bacteria in human saliva and in vitro biofilms with quantitative real-time PCR.

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    Price, R R; Viscount, H B; Stanley, M C; Leung, K-P

    2007-01-01

    An in vitro plaque model based on the use of human salivary bacteria and tooth-like surfaces was previously developed for studying the formation of oral biofilm and its use for pre-clinical testing of candidate antimicrobial or antiplaque agents. In this study, a quantitative Taqman PCR assay (QPCR) was developed to compare the bacterial compositions of in vitro biofilms to parent saliva samples, and to determine the relative contributions of different species in the formation of the oral biofilm. In addition, the growth inhibition of saliva-derived plaque was evaluated by chlorhexidine. With this assay, which consisted of primer/probe sets targeting either 16S rDNA sequences present in public databases or cloned ribosomal intergenic spacer region (ISR) sequences, 15 oral bacteria derived from saliva as well as those that were responsible for biofilm formation in an in vitro plaque model were rapidly identified and quantified. Among the target organisms were Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum, Lactobacillus acidophilus, Micromonas micros, Porphyromonas gingivalis, Prevotella intermedia, Streptococcus mutans, Streptococcus sobrinus, Tannerella forsythensis, and Veillonella parvula. Primer and probe sets developed were both sensitive and specific. The relative profiles of a number of bacteria in 45-h-old biofilms were determined and, when compared to saliva samples, it was found that most of the bacteria identified in saliva also populated the in vitro plaque, including some anaerobes. Brief exposure of biofilms to chlorhexidine resulted in significant losses in viability. This new broad spectrum QPCR assay in combination with the in vitro plaque model will be of significant value in the quantitative study of the microbial composition of human saliva, saliva-derived plaque, and pre-clinical evaluation of potential antimicrobial and antiplaque molecules.

  19. Risk factors associated with pleuritis and cranio-ventral pulmonary consolidation in slaughter-aged pigs.

    Science.gov (United States)

    Fraile, L; Alegre, A; López-Jiménez, R; Nofrarías, M; Segalés, J

    2010-06-01

    Examination of lung lesions at the slaughterhouse is a useful tool to estimate the importance of respiratory disease at farm, regional or national level. The objective of the present work was to describe the prevalence of gross lung lesions at slaughter, with a special focus on pleuritis and cranio-ventral pulmonary consolidation, and to identify major risk factors for these lesions. Data from 107 farms involving approximately 11,000 pigs enabled gross lung lesions to be correlated with serology to different swine respiratory pathogens as well as with production system characteristics and vaccination schedules. Pleuritis and cranio-ventral pulmonary consolidation lesions were recorded in 26.8% and 55.7% of slaughter-aged pigs, respectively. Among lungs with pleuritis, 50.1% had lesions compatible with Actinobacillus pleuropneumoniae (App) infection. Antibodies to porcine reproductive and respiratory syndrome (PRRSV), three subtypes (H1N1, H1N2 and H3N2) of swine influenza virus (SIV), App and Mycoplasma hyopneumoniae (Mhyo) were highly prevalent (>82%) in most of the farms. In a multivariable analysis, it was estimated (R(2)=0.40) that the percentage of animals with pleuritis compatible with App infection depended on the existence of an all in-all out by room management system and App and PRRSV herd seroprevalence. Moreover, it was possible to foresee (R(2)=0.59) that cranio-ventral pulmonary consolidation lesions (EP-like lesions) were affected by the type of farm ventilation, the presence of respiratory symptoms during the fattening period and Mhyo and SIV H1N2 herd seroprevalence.

  20. 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.

  1. Molecular Taxonomic Profiling of Bacterial Communities in a Gilthead Seabream (Sparus aurata) Hatchery

    Science.gov (United States)

    Califano, Gianmaria; Castanho, Sara; Soares, Florbela; Ribeiro, Laura; Cox, Cymon J.; Mata, Leonardo; Costa, Rodrigo

    2017-01-01

    As wild fish stocks decline worldwide, land-based fish rearing is likely to be of increasing relevance to feeding future human generations. Little is known about the structure and role of microbial communities in fish aquaculture, particularly at larval developmental stages where the fish microbiome develops and host animals are most susceptible to disease. We employed next-generation sequencing (NGS) of 16S rRNA gene reads amplified from total community DNA to reveal the structure of bacterial communities in a gilthead seabream (Sparus aurata) larviculture system. Early- (2 days after hatching) and late-stage (34 days after hatching) fish larvae presented remarkably divergent bacterial consortia, with the genera Pseudoalteromonas, Marinomonas, Acinetobacter, and Acidocella (besides several unclassified Alphaproteobacteria) dominating the former, and Actinobacillus, Streptococcus, Massilia, Paracoccus, and Pseudomonas being prevalent in the latter. A significant reduction in rearing-water bacterial diversity was observed during the larviculture trial, characterized by higher abundance of the Cryomorphaceae family (Bacteroidetes), known to populate microniches with high organic load, in late-stage rearing water in comparison with early-stage rearing-water. Furthermore, we observed the recruitment, into host tissues, of several bacterial phylotypes—including putative pathogens as well as mutualists—that were detected at negligible densities in rearing-water or in the live feed (i.e., rotifers and artemia). These results suggest that, besides host-driven selective forces, both the live feed and the surrounding rearing environment contribute to shaping the microbiome of farmed gilthead sea-bream larvae, and that a differential establishment of host-associated bacteria takes place during larval development. PMID:28261166

  2. Effects of eliminating pyruvate node pathways and of coexpression of heterogeneous carboxylation enzymes on succinate production by Enterobacter aerogenes.

    Science.gov (United States)

    Tajima, Yoshinori; Yamamoto, Yoko; Fukui, Keita; Nishio, Yousuke; Hashiguchi, Kenichi; Usuda, Yoshihiro; Sode, Koji

    2015-02-01

    Lowering the pH in bacterium-based succinate fermentation is considered a feasible approach to reduce total production costs. Newly isolated Enterobacter aerogenes strain AJ110637, a rapid carbon source assimilator under weakly acidic (pH 5.0) conditions, was selected as a platform for succinate production. Our previous work showed that the ΔadhE/PCK strain, developed from AJ110637 with inactivated ethanol dehydrogenase and introduced Actinobacillus succinogenes phosphoenolpyruvate carboxykinase (PCK), generated succinate as a major product of anaerobic mixed-acid fermentation from glucose under weakly acidic conditions (pH <6.2). To further improve the production of succinate by the ΔadhE/PCK strain, metabolically engineered strains were designed based on the elimination of pathways that produced undesirable products and the introduction of two carboxylation pathways from phosphoenolpyruvate and pyruvate to oxaloacetate. The highest production of succinate was observed with strain ES04/PCK+PYC, which had inactivated ethanol, lactate, acetate, and 2,3-butanediol pathways and coexpressed PCK and Corynebacterium glutamicum pyruvate carboxylase (PYC). This strain produced succinate from glucose with over 70% yield (gram per gram) without any measurable formation of ethanol, lactate, or 2,3-butanediol under weakly acidic conditions. The impact of lowering the pH from 7.0 to 5.5 on succinate production in this strain was evaluated under pH-controlled batch culture conditions and showed that the lower pH decreased the succinate titer but increased its yield. These findings can be applied to identify additional engineering targets to increase succinate production.

  3. High-Throughput Identification of Bacteria and Yeast by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry in Conventional Medical Microbiology Laboratories ▿

    Science.gov (United States)

    van Veen, S. Q.; Claas, E. C. J.; Kuijper, Ed J.

    2010-01-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory. PMID:20053859

  4. 猪传染性胸膜肺炎在保育猪中的流行病学调查%Epidemiological Survey on Nursery Pig of Porcine Contagious Pleuropneu-monia

    Institute of Scientific and Technical Information of China (English)

    谢艳霞; 王重龙; 吴东; 周学利; 沈学怀; 赵瑞宏; 胡晓苗; 侯宏艳; 戴银; 潘孝成; 张丹俊

    2016-01-01

    [Objective]To investigate the infection rates of Actinobacillus pleuropneumoniae on piglets in large and medium scale pig farms. [Method] Total of 272 piglets sera collected from 10 different swine farms were detected for the antibody against APP by using ApxIV-ELISA method. [Result]The re-sults showed that the total positive rates of A.pleuropneumoniae in all detected pig samples were 32.4%. [Conclusion] These results indicated that A.pleurop-neumoniae was very common in part of scale pig farms. The infection rates of A. pleuropneumoniae on piglets in large and medium scale pig farms were high and no obvious regularity. The infection rates of piglets were not related to incidence of disease. The liquidity of the pigs out-side had little effect on the A. pleurop-neumoniae infection rates in farms.%[目的]调查大、中型规模种猪场保育猪胸膜肺炎放射杆菌(APP)感染情况。[方法]用ApxIV-ELISA 试剂盒检测272份保育猪血清。[结果]有88份猪血清样品检测到APP感染抗体阳性,阳性率为32.4%。[结论]APP普遍存在于各猪场,大、中型种猪场保育猪APP 感染抗体阳性率偏高且无明显规律,保育猪APP 的感染率与猪只是否发病无关,场外猪群的流动性对大型猪场内APP的感染率影响不大。

  5. Clinical and microbiological effects of controlled-release locally delivered minocycline in periodontitis.

    Science.gov (United States)

    Jones, A A; Kornman, K S; Newbold, D A; Manwell, M A

    1994-11-01

    The clinical efficacy of minocycline in a subgingival local delivery system was evaluated alone (M) or as an adjunct to scaling and root planing (M + SRP), in comparison to scaling and root planing (SRP) or to no subgingival treatment (NoTx) in adult periodontitis. Fifty-one adult patients with > or = 7 mm periodontal pockets demonstrating the presence by culture of Porphyromonas gingivalis (Pg), Prevotella intermedia (P(i)), or Actinobacillus actinomycetemcomitans (Aa) were randomized into one of the above 4 treatment groups. All sites > or = 5 mm in the most diseased quadrant in each patient received the therapy. Other quadrants were not treated. All patients received standardized oral hygiene instructions at the beginning of the study. At 0, 1, 3 and 6 months following therapy the 7 mm experimental sites were evaluated for selected periodontal pathogens by DNA probe analysis. At these same time points, the plaque index, gingival index, and bleeding on probing were evaluated as well as probing depth and relative clinical attachment level which were assessed by means of an automated probe. Probing depth reduction with M + SRP was significantly greater than all other groups at one month and significantly greater than NoTx and SRP at 3 months. There were no differences in probing depth reduction among groups at 6 months. At 6 months the gain in clinical attachment level was significantly greater for SRP than for either the NoTx or M groups. The prevalence of Pg decreased significantly in the M and M + SRP groups at one month.(ABSTRACT TRUNCATED AT 250 WORDS)

  6. Florfenicol - pharmacodynamic, pharmacokinetics and clinical efficacy of oral formulations in domestic animals: A systematic review

    Directory of Open Access Journals (Sweden)

    Ščuka Leon

    2005-01-01

    Full Text Available Porcine respiratory disease complex (PRDC is a major economic problem for swine producers world-wide. Pharmacodynamic, pharmacokinetics and clinical efficacy of florfenicol oral formulations in domestic animals were evaluated. For this purpose the systematic review and meta-analysis were done. In vitro efficacy of florfenicol showed that this drug is highly effective against most important respiratory pathogens. All these facts are shown in our survey. Three studies in pigs were relevant to include in the meta-analysis, which showed that results in the florfenicol group were better than in comparative control groups in all observed parameters: clinical signs, lung lesions and resolution of Actinobacillus pleuropneumoniae (P<0,001. A second meta-analysis with 7 studies showed that the usage of florfenicol reduces mortality in pig herds with PRDC (P<0.05. Other field trials in pigs using florfenicol oral forms where reviewed. After treatment with florfenicol oral solution there was a significant drop of mortality in both groups of pigs (P<0.01; eg. one using florfenicol oral solution in treating PRDC (n=85 and another mixed pneumoenteric infection (n=54. Analysis of data when using premix in pigs (n=118 also suggests that a medicated premix has a favorable anti-infectious effect on pigs, irrespective of the group of animals or the evolution stage of the disease. Finally, favorable effect of florfenicol in treating swine ileitis was also presented. Regarding their pharmacokinetics, in vitro and clinical efficacy of florfenicol oral forms, they should be considered as a powerful tool for combating complex infections that are frequently met in intensive animal production.

  7. REVIEW paper: mare reproductive loss syndrome.

    Science.gov (United States)

    Sebastian, M M; Bernard, W V; Riddle, T W; Latimer, C R; Fitzgerald, T D; Harrison, L R

    2008-09-01

    An epidemic of early fetal loss (EFL), late fetal loss (LFL), fibrinous pericarditis, and unilateral uveitis which occurred during the spring of 2001, are together now known as the mare reproductive loss syndrome (MRLS). A similar epidemic with less intensity was reported during the same period of time from southern Ohio, West Virginia, and Tennessee. The same syndrome with lesser intensity recurred in 2002. The estimated economic loss from the syndrome in 2001 and 2002 together was approximately $500 million. Both EFL and LFL were characterized by the absence of specific clinical signs in aborting mares. Nonhemolytic Streptococcus spp. and Actinobacillus spp. accounted for 65% of the organisms isolated from fetuses submitted for a postmortem during the MRLS period in 2001 and 2002. The pathologic findings in fetoplacental units of LFL included bronchopneumonia and funisitis, and there were no findings in EFL. Epidemiologic studies conducted in 2001 suggested an association between the presences of eastern tent caterpillars (ETC) in pastures with MRLS. Experimental studies in pregnant mares by exposure to ETC, or administration by stomach tube or with feed material, reproduced EFL and LFL. Similar experimental studies in mouse, rats, and goats with ETC were unsuccessful. Currently, 2 hypotheses are proposed for MRLS. One hypothesis proposes that an ETC-related toxin with secondary opportunistic bacterial invasion of the fetus leads to MRLS. The second hypothesis suggests that a breach of gastrointestinal mucosal integrity by hairs of ETC leads to a bacteremia and results in MRLS. In 2004, a similar equine abortion storm was reported from Australia and caterpillar exposure was identified as a risk factor for the abortion. In 2006, the syndrome was observed in Florida and New Jersey.

  8. Susceptibility of bacteria isolated from pigs to tiamulin and enrofloxacin metabolites.

    Science.gov (United States)

    Lykkeberg, Anne Kruse; Halling-Sørensen, Bent; Jensen, Lars Bogø

    2007-03-31

    Susceptibilities to metabolites of tiamulin (TIA) and enrofloxacin (ENR) were tested using selected bacteria with previously defined minimal inhibitory concentrations (MIC). The TIA metabolites tested were: N-deethyl-tiamulin (DTIA), 2beta-hydroxy-tiamulin (2beta-HTIA) and 8alpha-hydroxy-tiamulin (8alpha-HTIA), and the ENR metabolites were: ciprofloxacin (CIP) and enrofloxacin N-oxide (ENR-N). Bacteria, all of porcine origin, were selected as representatives of bacterial infections (Staphylococcus hyicus and Actinobacillus pleuropneumoniae), zoonotic bacteria (Campylobacter coli) and indicator bacteria (Escherichia coli and enterococci). Furthermore the effects of these compounds were tested on the microbial community of active sludge to test any negative effect on colony forming units (CFU). DTIA had a potency of 12.5-50% of the potency of TIA. 2beta-HTIA and 8alpha-HTIA had potencies less than 1% of the potency of TIA. ENR-N had a potency of 0.75-1.5% of the potency of ENR, while CIP and ENR had similar potencies. Results obtained here indicate that CIP and DTIA could contribute to the selective pressure for upholding antimicrobial resistant bacteria in animals under ENR or TIA treatment. The most potent metabolites CIP and DTIA showed considerable potencies against activated sludge bacteria compared to the parent compounds. EC(50) (microg/ml) for ENR, CIP, TIA and DTIA were 0.018 [95% CI: 0.028-0.149], 0.064 [95% CI: 0.007-0.046], 6.0 [95% CI: 3.6-9.8], and 9.7 [95% CI: 5.8-16.3], respectively. This indicates that the compounds can change the bacterial population in the sludge, and hereby alter the properties of the sludge.

  9. In vitro activity and rodent efficacy of clinafloxacin for bovine and swine respiratory disease.

    Science.gov (United States)

    Sweeney, Michael T; Quesnell, Rebecca; Tiwari, Raksha; Lemay, Mary; Watts, Jeffrey L

    2013-01-01

    Clinafloxacin is a broad-spectrum fluoroquinolone that was originally developed and subsequently abandoned in the late 1990s as a human health antibiotic for respiratory diseases. The purpose of this study was to investigate the activity of clinafloxacin as a possible treatment for respiratory disease in cattle and pigs. Minimum inhibitory concentration (MIC) values were determined using Clinical and Laboratory Standards Institute recommended procedures with recent strains from the Zoetis culture collection. Rodent efficacy was determined in CD-1 mice infected systemically or intranasally with bovine Mannheimia haemolytica or Pasteurella multocida, or swine Actinobacillus pleuropneumoniae, and administered clinafloxacin for determination of ED50 (efficacious dose-50%) values. The MIC90 values for clinafloxacin against bovine P. multocida, M. haemolytica, Histophilus somni, and M. bovis were 0.125, 0.5, 0.125, and 1 μg/ml, respectively, and the MIC90 values against swine P. multocida, A. pleuropneumoniae, S. suis, and M. hyopneumoniae were í0.03, í0.03, 0.125, and í0.008 μg/ml, respectively. Efficacy in mouse models showed average ED50 values of 0.019 mg/kg/dose in the bovine M. haemolytica systemic infection model, 0.55 mg/kg in the bovine P. multocida intranasal lung challenge model, 0.08 mg/kg/dose in the bovine P. multocida systemic infection model, and 0.7 mg/kg/dose in the swine A. pleuropneumoniae systemic infection model. Clinafloxacin shows good in vitro activity and efficacy in mouse models and may be a novel treatment alternative for the treatment of respiratory disease in cattle and pigs.

  10. Monitoring of antimicrobial susceptibility of respiratory tract pathogens isolated from diseased cattle and pigs across Europe, 2009-2012: VetPath results.

    Science.gov (United States)

    El Garch, Farid; de Jong, Anno; Simjee, Shabbir; Moyaert, Hilde; Klein, Ulrich; Ludwig, Carolin; Marion, Hervé; Haag-Diergarten, Silke; Richard-Mazet, Alexandra; Thomas, Valérie; Siegwart, Ed

    2016-10-15

    VetPath is an ongoing pan-European antibiotic susceptibility monitoring programme that collects pathogens from diseased cattle, pigs and poultry. In the current study, 996 isolates from cattle and pig respiratory tract infections were tested for their antimicrobial susceptibilities. Non-replicate lung samples or nasopharyngeal/nasal swabs were collected from animals with acute clinical signs in 10 countries during 2009-2012. Pasteurella multocida, Mannheimia haemolytica and Histophilus somni from cattle and P. multocida, Actinobacillus pleuropneumoniae, Haemophilus parasuis, Bordetella bronchiseptica and Streptococcus suis from pigs were isolated by standard methods. S. suis was also isolated from meningitis cases. MIC values of 16 or 17 antibiotics were assessed centrally by broth microdilution following CLSI standards. Results were interpreted using CLSI breakpoints where available. Cattle isolates were generally highly susceptible to most antibiotics, except to tetracycline (3.0-12.0% resistance). Low levels of resistance (0-4.0%) were observed for the macrolide antibiotics. Resistance to spectinomycin varied from 0 to 6.0%. In pig isolates similar observations were made. Resistance to amoxicillin/clavulanic acid, ceftiofur, enrofloxacin, florfenicol, tulathromycin, tiamulin and tilmicosin was absent or <2%. Trimethoprim/sulfamethoxazole resistance varied from 1.9 to 5.3%, but tetracycline resistance varied from 20.4% in P. multocida to 88.1% in S. suis. For most antibiotics and pathogens the percentage resistance remained unchanged or only increased numerically as compared to that of the period 2002-2006. In conclusion, absence or low resistance to antibiotics with defined clinical breakpoints, except for tetracycline, was observed among the major respiratory tract pathogens recovered from livestock. Comparison of all antibiotics and organisms was hampered since for almost half of the antibiotics no CLSI-defined breakpoints were available.

  11. Plasma concentrations resulting from florfenicol preparations given to pigs in their drinking water.

    Science.gov (United States)

    Gutiérrez, L; Vargas, D; Ocampo, L; Sumano, H; Martinez, R; Tapia, G

    2011-09-01

    Florfenicol administered through the drinking water has been recommended as a metaphylactic antibacterial drug to control outbreaks of respiratory diseases in pigs caused by strains of Actinobacillus pleuropneumoniae and Pasteurella multocida, yet it is difficult to pinpoint in practice when the drug is given metaphylactically or therapeutically. Further, pigs are likely to reject florfenicol-medicated water, and plasma concentrations of the drug are likely to be marginal for diseases caused by Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. The reported minimal inhibitory concentration (MIC) values for these organisms show a breakpoint of 2 to 3 μg/mL. An experiment was conducted during September and October 2009. One hundred twenty healthy crossbred pigs (Landrace-Yorkshire), weighing 23 ± 6.2 kg, were used in this trial. They were randomly assigned to 5 groups, with 3 replicates of 8 animals/group. Two commercial preparations of florfenicol were administered through the drinking water at 2 concentrations (0.01 and 0.015%). Water intake was measured before and after medication, and plasma concentrations of florfenicol were determined by HPLC. Considerable rejection of florfenicol-medicated water was observed. However, plasma florfenicol concentrations were of a range sufficient for a methaphylaxis approach to preventing disease by bacteria, with MIC breakpoints of ≤ 0.25 μg/mL. Decreased efficacy as a metaphylactic medication should be expected for bacteria with MIC >0.25 μg/mL, considering the reported existence of bacteria resistant to florfenicol and the natural resistance of Streptococcus suis or E. coli to this drug.

  12. Severity of Bovine Tuberculosis Is Associated with Co-Infection with Common Pathogens in Wild Boar

    Science.gov (United States)

    Risco, David; Serrano, Emmanuel; Fernández-Llario, Pedro; Cuesta, Jesús M.; Gonçalves, Pilar; García-Jiménez, Waldo L.; Martínez, Remigio; Cerrato, Rosario; Velarde, Roser; Gómez, Luis; Segalés, Joaquím; Hermoso de Mendoza, Javier

    2014-01-01

    Co-infections with parasites or viruses drive tuberculosis dynamics in humans, but little is known about their effects in other non-human hosts. This work aims to investigate the relationship between Mycobacterium bovis infection and other pathogens in wild boar (Sus scrofa), a recognized reservoir of bovine tuberculosis (bTB) in Mediterranean ecosystems. For this purpose, it has been assessed whether contacts with common concomitant pathogens are associated with the development of severe bTB lesions in 165 wild boar from mid-western Spain. The presence of bTB lesions affecting only one anatomic location (cervical lymph nodes), or more severe patterns affecting more than one location (mainly cervical lymph nodes and lungs), was assessed in infected animals. In addition, the existence of contacts with other pathogens such as porcine circovirus type 2 (PCV2), Aujeszky's disease virus (ADV), swine influenza virus, porcine reproductive and respiratory syndrome virus, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, Haemophilus parasuis and Metastrongylus spp, was evaluated by means of serological, microbiological and parasitological techniques. The existence of contacts with a structured community of pathogens in wild boar infected by M. bovis was statistically investigated by null models. Association between this community of pathogens and bTB severity was examined using a Partial Least Squares regression approach. Results showed that adult wild boar infected by M. bovis had contacted with some specific, non-random pathogen combinations. Contact with PCV2, ADV and infection by Metastrongylus spp, was positively correlated to tuberculosis severity. Therefore, measures against these concomitant pathogens such as vaccination or deworming, might be useful in tuberculosis control programmes in the wild boar. However, given the unexpected consequences of altering any community of organisms, further research should evaluate the impact of such measures under

  13. Optimal combinations of acute phase proteins for detecting infectious disease in pigs

    Directory of Open Access Journals (Sweden)

    Heegaard Peter MH

    2011-03-01

    Full Text Available Abstract The acute phase protein (APP response is an early systemic sign of disease, detected as substantial changes in APP serum concentrations and most disease states involving inflammatory reactions give rise to APP responses. To obtain a detailed picture of the general utility of porcine APPs to detect any disease with an inflammatory component seven porcine APPs were analysed in serum sampled at regular intervals in six different experimental challenge groups of pigs, including three bacterial (Actinobacillus pleuropneumoniae, Streptococcus suis, Mycoplasma hyosynoviae, one parasitic (Toxoplasma gondii and one viral (porcine respiratory and reproductive syndrome virus infection and one aseptic inflammation. Immunochemical analyses of seven APPs, four positive (C-reactive protein (CRP, haptoglobin (Hp, pig major acute phase protein (pigMAP and serum amyloid A (SAA and three negative (albumin, transthyretin, and apolipoprotein A1 (apoA1 were performed in the more than 400 serum samples constituting the serum panel. This was followed by advanced statistical treatment of the data using a multi-step procedure which included defining cut-off values and calculating detection probabilities for single APPs and for APP combinations. Combinations of APPs allowed the detection of disease more sensitively than any individual APP and the best three-protein combinations were CRP, apoA1, pigMAP and CRP, apoA1, Hp, respectively, closely followed by the two-protein combinations CRP, pigMAP and apoA1, pigMAP, respectively. For the practical use of such combinations, methodology is described for establishing individual APP threshold values, above which, for any APP in the combination, ongoing infection/inflammation is indicated.

  14. An approach to a patient with infective endocarditis.

    Science.gov (United States)

    Hitzeroth, J; Beckett, N; Ntuli, P

    2016-02-01

    Although infective endocarditis (IE) is relatively uncommon, it remains an important clinical entity with a high in-hospital and 1-year mortality. It is most commonly caused by viridans streptococci. Staphylococcus aureus is responsible for a malignant course of IE and often requires early surgery to eradicate. Other rarer causes are various bacilli, including the HACEK (Haemophilus, Actinobacillus,Cardiobacterium, Eikenella and Kingella spp.) group of organisms and fungi. The clinical presentation varies. Patients may present with a nonspecific illness, valve dysfunction, heart failure (HF) and symptoms due to peripheral embolisation. The diagnosis is traditionally based on the modified Duke criteria and rests mainly on clinical features and to a lesser extent on certain laboratory findings,microbiological assessment and cardiovascular imaging. Identification of the offending micro-organism is not only important from a diagnostic point of view, but also makes targeted antibiotic treatment possible and provides useful prognostic information. A significant proportion of microbiological cultures are negative, frequently owing to the administration of antibiotics prior to appropriate culture.Blood-culture-negative IE poses significant diagnostic and treatment challenges. The course of the disease is frequently complicated, and sequelae include HF, local intracardiac extension of infection (abscess, fistula, pseudoaneurysm), stroke and intracranial haemorrhage due to septic emboli or mycotic aneurysm formation as well as renal injury. Management includes prolonged intravenous antibiotics and consideration for early surgery with removal of infective tissue and valve replacement in patients who have poor prognostic features or complications. Antibiotic administration for at-risk patients to prevent bacteraemia during specific procedures (particularly dental) is recommended to prevent IE. The patient population who would benefit from antibiotic prophylaxis has become

  15. Detection of periodontopathogenic bacteria in pregnant women by traditional anaerobic culture method and by a commercial molecular genetic method.

    Science.gov (United States)

    Urbán, Edit; Terhes, Gabriella; Radnai, Márta; Gorzó, István; Nagy, Elisabeth

    2010-06-01

    To culture facultative and strict anaerobic bacteria is a well-established method for analyzing subgingival plaque samples. Micro-IDent and micro-IDent Plus (HAIN Lifescience GmbH, Nehren, Germany) tests are two commercially available rapid PCR-based methods for the identification and quantification of putative periodontopathogen bacteria. In this study, we compared these commercial PCR-based hybridization methods with conventional anaerobic culture technique. A total of 36 subgingival plaque samples were collected from periodontal pockets of pregnant women with chronic localized periodontitis. Aliquots of these samples were evaluated with species-specific probes provided by micro-IDent and micro-IDent Plus tests simultaneously, and from the same samples anaerobic and capnophylic bacteria were cultured on selective media. The overall agreement between both methods was excellent for Eubacterium nodatum, Tannerella forsythia and Porphyromonas gingivalis (97-92%), fair for Capnocytophaga sp, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Prevotella intermedia (91-89%) and poor for Fusobacterium nucleatum, Parvimonas micra (Micromonas micros), and Campylobacter rectus (86-78%). Discrepancies in the results may be explained by inability of culture method to distinguish between closely related taxa (e.i P. intermedia/Prevotella. nigrescens), and problems of keeping periodontopathogen bacteria viable, which is required for successful detection by standard culture method. Nucleic acid-based methods may replace cultivation method as frequently used methods in microbiological diagnosis of progressive periodontitis, thus micro-IDent and micro-IDent Plus tests can be recommended where culture of periodontopathogenic bacteria is not performed in routine microbiology laboratories to analyze subgingival plaque samples.

  16. Association of swine influenza H1N1 pandemic virus (SIV-H1N1p) with porcine respiratory disease complex in sows from commercial pig farms in Colombia.

    Science.gov (United States)

    Jiménez, Luisa Fernanda Mancipe; Ramírez Nieto, Gloria; Alfonso, Victor Vera; Correa, Jairo Jaime

    2014-08-01

    Porcine respiratory disease complex (PRDC) is a serious health problem that mainly affects growing and finishing pigs. PRDC is caused by a combination of viral and bacterial agents, such as porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), Mycoplasma hyopneumoniae (Myh), Actinobacillus pleuropneumoniae (APP), Pasteurella multocida and Porcine circovirus 2 (PCV2). To characterize the specific role of swine influenza virus in PRDC presentation in Colombia, 11 farms from three major production regions in Colombia were examined in this study. Nasal swabs, bronchial lavage and lung tissue samples were obtained from animals displaying symptoms compatible with SIV. Isolation of SIV was performed in 9-day embryonated chicken eggs or Madin-Darby Canine Kidney (MDCK) cells. Positive isolates, identified via the hemagglutination inhibition test, were further analyzed using PCR. Overall, 7 of the 11 farms were positive for SIV. Notably, sequencing of the gene encoding the hemagglutinin (HA) protein led to grouping of strains into circulating viruses identified during the human outbreak of 2009, classified as pandemic H1N1-2009. Serum samples from 198 gilts and multiparous sows between 2008 and 2009 were obtained to determine antibody presence of APP, Myh, PCV2 and PRRSV in both SIV-H1N1p-negative and -positive farms, but higher levels were recorded for SIV-H1N1p-positive farms. Odds ratio (OR) and P values revealed statistically significant differences (p<0.05) in PRDC presentation in gilts and multiparous sows of farms positive for SIV-H1N1p. Our findings indicate that positive farms have increased risk of PRDC presentation, in particular, PCV2, APP and Myh.

  17. Putative periodontopathogens in "diseased" and "non-diseased" persons exhibiting poor oral hygiene.

    Science.gov (United States)

    Dahlén, G; Manji, F; Baelum, V; Fejerskov, O

    1992-01-01

    The aim of the study was to assess the occurrence of some putative periodonto-pathogens in "test" and "control" sites in "diseased" and "non-diseased" persons, respectively, from an adult rural Kenyan population exhibiting poor oral hygiene and widespread loss of attachment (LA). 14 persons (less than 35 years) were assigned to a "diseased" category on the basis of at least 4 sites with LA greater than or equal to 4 mm; at least 5 mm LA and a pocket greater than or equal to 4 mm interproximally in a lower incisor ("test" site): and less than 2 mm LA and no pocket greater than or equal to 4 mm distal to a lower canine or mesial to a lower first premolar ("control" site). Age-matched "non-diseased" persons were identified on the basis of no sites with LA greater than 2 mm and no pockets greater than or equal to 4 mm associated with LA. Paperpoint samples from test and control sites as well as a scraping sample from the dorsum of tongue were examined for presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Bacteroides intermedius, B. melaninogenicus group, Capnocytophaga, Selenomonas spp., and Wolinella recta. P. gingivalis was found in 79% of test sites and 36% of control sites in "diseased" persons, and in 18% and 35% of test and control sites, respectively, in "non-diseased" persons. "No other bacterial group discriminated significantly between test and control sites or between diseased and non-diseased subjects. The surprisingly high occurrence of P. gingivalis in non-diseased subjects, both subgingivally and on tongue, indicates that deep periodontal pockets are not prerequisite ecological environments for P. gingivalis establishment.

  18. Association between selected oral pathogens and gastric precancerous lesions.

    Directory of Open Access Journals (Sweden)

    Christian R Salazar

    Full Text Available We examined whether colonization of selected oral pathogens is associated with gastric precancerous lesions in a cross-sectional study. A total of 119 participants were included, of which 37 were cases of chronic atrophic gastritis, intestinal metaplasia, or dysplasia. An oral examination was performed to measure periodontal indices. Plaque and saliva samples were tested with real-time quantitative PCR for DNA levels of pathogens related to periodontal disease (Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, Actinobacillus actinomycetemcomitans and dental caries (Streptococcus mutans and S. sobrinus. There were no consistent associations between DNA levels of selected bacterial species and gastric precancerous lesions, although an elevated but non-significant odds ratio (OR for gastric precancerous lesions was observed in relation to increasing colonization of A. actinomycetemcomitans (OR = 1.36 for one standard deviation increase, 95% Confidence Interval = 0.87-2.12, P. gingivalis (OR = 1.12, 0.67-1.88 and T. denticola (OR = 1.34, 0.83-2.12 measured in plaque. To assess the influence of specific long-term infection, stratified analyses by levels of periodontal indices were conducted. A. actinomycetemcomitans was significantly associated with gastric precancerous lesions (OR = 2.51, 1.13-5.56 among those with ≥ median of percent tooth sites with PD ≥ 3 mm, compared with no association among those below the median (OR = 0.86, 0.43-1.72. A significantly stronger relationship was observed between the cumulative bacterial burden score of periodontal disease-related pathogens and gastric precancerous lesions among those with higher versus lower levels of periodontal disease indices (p-values for interactions: 0.03-0.06. Among individuals with periodontal disease, high levels of colonization of periodontal pathogens are associated with an increased risk of gastric precancerous lesions.

  19. Antibacterial activity and cell compatibility of TiZrN, TiZrCN, and TiZr-amorphous carbon coatings

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Heng-Li [School of Dentistry, China Medical University, Taichung 404, Taiwan (China); Department of Bioinformatics and Medical Engineering, Asia University, Taichung 41354, Taiwan (China); Chang, Yin-Yu, E-mail: yinyu@nfu.edu.tw [Department of Mechanical and Computer-aided Engineering, National Formosa University, Yunlin 632, Taiwan (China); Liu, Jia-Xu [Department of Mechanical and Computer-aided Engineering, National Formosa University, Yunlin 632, Taiwan (China); Tsai, Ming-Tzu [Department of Biomedical Engineering, Hungkuang University, Taichung 433, Taiwan (China); Lai, Chih-Ho [Department of Microbiology and Immunology, Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan, Taiwan (China)

    2015-12-01

    A cathodic-arc evaporation system with plasma-enhanced duct equipment was used to deposit TiZrN, TiZrCN, and TiZr/a-C coatings. Reactive gases (N{sub 2} and C{sub 2}H{sub 2}) activated by the Ti and Zr plasma in the evaporation process was used to deposit the TiZrCN and TiZr/a-C coatings with different C and nitrogen contents. The crystalline structures and bonding states of coatings were analyzed by X-ray diffraction and X-ray photoelectron spectroscopy. The microbial activity of the coatings was evaluated against Staphylococcus aureus (Gram-positive bacteria) and Actinobacillus actinomycetemcomitans (Gram-negative bacteria) by in vitro antibacterial analysis using a fluorescence staining method employing SYTO9 and a bacterial-viability test on an agar plate. The cell compatibility and morphology related to CCD-966SK cell-line human skin fibroblast cells on the coated samples were also determined using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, reverse-transcriptase-polymerase chain reaction, and scanning electron microscopy. The results suggest that the TiZrCN coatings not only possess better antibacterial performance than TiZrN and TiZr/a-C coatings but also maintain good compatibility with human skin fibroblast cells. - Highlights: • TiZrN, TiZrCN, and TiZr/a-C coatings were deposited using cathodic arc evaporation. • The TiZrCN showed a composite structure containing TiN, ZrN, and a-C. • The TiZrCN-coated Ti showed the least hydrophobicity among the samples. • The TiZrCN-coated Ti showed good human skin fibroblast cell viability. • The TiZrCN-coated Ti exhibited good antibacterial performance.

  20. 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.

  1. High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization-time of flight mass spectrometry in conventional medical microbiology laboratories.

    Science.gov (United States)

    van Veen, S Q; Claas, E C J; Kuijper, Ed J

    2010-03-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory.

  2. Preparation of saccharification liquid for succinic acid by fermentation from corncob sulfuric acid hydrolysis%稀酸水解玉米芯制备丁二酸

    Institute of Scientific and Technical Information of China (English)

    姚嘉旻; 姜岷; 吴昊; 陈可泉

    2010-01-01

    利用正交设计得到稀H2SO4水解玉米芯制备混合糖液的优化工艺:玉米芯料液比1:5(质量体积比),物料粒径250~380 μm、H2SO4用量3%(体积分数)、水解温度126 ℃、反应时间2.5 h.此工艺条件下的总糖收率达90%,总糖质量浓度为60 g/L,发酵抑制物糠醛含量为0.87 g/L,5羟甲基糠醛含量为0.68 g/L.在此基础上利用活性炭吸附和Ca(OH)2中和对玉米芯混合糖液进行脱毒及脱盐处理,SO2-4脱除率达96%,色素脱除率为96%,糠醛、5羟甲基糠醛及多酚类物质脱除率均高于50%.处理后的玉米芯多组分糖液作为产琥珀酸放线杆菌(Actinobacillus succinogenes)NJ113的发酵C源,当培养基中初始总糖质量浓度为50 g/L时,丁二酸收率为61.68%,丁二酸质量浓度为30.8 g/L;初始总糖质量浓度为70 g/L时,丁二酸收率仍可达50%以上,丁二酸质量浓度为35.2 g/L.发酵实验表明,将经过脱毒脱盐处理的玉米芯多组分糖液替代葡萄糖作为C源发酵制备丁二酸具有可行性.

  3. Relationship between herpesviruses and periodontal pathogenic bacteria in subgingival plaque%侵袭性牙周炎患者龈下菌斑中的疱疹病毒与牙周致病菌

    Institute of Scientific and Technical Information of China (English)

    丁芳; 冯向辉; 孟焕新; 赵亦兵; 张立; 路瑞芳; 陈智滨

    2008-01-01

    目的:检测侵袭性牙周炎(aggressive periodontitis, AgP)患者龈下菌斑中的人巨细胞病毒(human cytomegalovirus, HCMV)、EB病毒1型(Epstein-Barr virus.1,EBV一1),探讨疱疹病毒与牙周致病菌及牙周临床指标间的关系.方法:应用巢式PCR技术,检测89名AgP患者和31名健康对照者龈下菌斑中的2种疱疹病毒:人巨细胞病毒(HCMV)、EB病毒1型(EBV-1),同时应用16S rRNA为基础的PCR技术检测8种牙周可疑致病菌,包括伴放线放线杆菌(Actinobacillus actinomycetemcomitans, Aa)、牙龈卟啉单胞菌(Porphyromonas gingivalis, Pg)、福赛类杆菌(Bacteroides forsythus, Bf;现已更名为Tannerella forsythensis, Tf)、中间普氏菌(Prevotella intermedia, Pi)、直形弯曲菌(Campylobacter rectus, Cr)、具核梭杆菌(Fusobacterium nucleatum, Fn)、牙密螺旋体(Treponema denticola, Td)、变黑普氏菌(Prevotella nigrescens, Pn).结果:89名AgP患者的HCMV检出率为43.8%,显著高于健康对照者(12.9%),差异有统计学意义(P0.05).检出6~8种牙周致病菌的AgP患者其HCMV和EBV-1检出率均明显高于仅检出3~5种细菌的AgP患者的病毒检出率(P<0.05).结论:HCMV、EBV-1在AgP患者龈下的检出率较健康对照高;且在检出牙周致病菌种类数较多的样本中,疱疹病毒的检出率高,提示牙周感染HCMV和EBV-1与侵袭性牙周炎相关,疱疹病毒与牙周致病菌在侵袭性牙周炎的发病中可能起着协同作用.

  4. Preparation, characterization and pharmacokinetics of doxycycline hydrochloride and florfenicol polyvinylpyrroliddone microparticle entrapped with hydroxypropyl-β-cyclodextrin inclusion complexes suspension.

    Science.gov (United States)

    Li, Xianqiang; Xie, Shuyu; Pan, Yuanhu; Qu, Wei; Tao, Yanfei; Chen, Dongmei; Huang, Lingli; Liu, Zhenli; Wang, Yulian; Yuan, Zonghui

    2016-05-01

    In order to effectively control the bacterial pneumonia in pigs, doxycycline hydrochloride (DoxHcl) and florfenicol (FF) microparticle suspension together with inclusion complexes was prepared by using hydroxypropyl-β-cyclodextrin (HP-β-CD) as host molecules, polyvinylpyrroliddone (PVP) as polymer carriers and hydroxypropyl methyl cellulose (HPMC) as suspending agents. In vitro antibacterial activity, properties, stability and pharmacokinetics of the suspension were studied. The results demonstrated that DoxHcl and FF had a synergistic or additive antibacterial activity against Streptococcus suis, Actinobacillus pleuropneumoniae and Haemophilus parasuis. The size, polydispersity index and zeta potential of microparticles were 1.46 ± 0.06 μm, 0.30 ± 0.02 and 1.53 ± 0.04 mV, respectively. The encapsulation efficiency (EE) of DoxHcl and FF was 45.28% ± 3.30% and 89.69% ± 2.71%, respectively. The re-dispersed time and sedimentation rate of the suspension were 1 min and 1. The suspension went through the 9-gage needle smoothly with withdrawal volume of 9.12 ± 0.87 mL/min. The suspension showed good stability when stored away from light, no irritation at the injection site and sustained release in PBS buffer. After intramuscular administration to pig, DoxHcl and FF could maintain over 0.15 μg/mL for 72 h. Compared to the control injection, the suspension increased the elimination half-life (T½ke) as well as mean residence time (MRT) of DoxHcl from 5.73 to 9.77 h and from 12.02 to 18.81 h, and those of FF from 12.02 to 26.19 h and from 12.02 to 28.16 h, respectively. The suspension increased the bioavailability of DoxHcl and FF by 1.74 and 1.13-fold, respectively. These results suggest that the compound suspension is a promising formulation for pig pneumonia therapy.

  5. 屠宰生猪胸膜炎的病原检测%Detection of Pleurisy Pathogens in Slaughter Swine

    Institute of Scientific and Technical Information of China (English)

    邱渊皓; 魏江华; 任娟娟; 唐攀; 刘万华; 武宁; 李涛; 伍成奇; 王晶钰

    2014-01-01

    Different degree of pleuritis lesions could usually be found in slaughter pigs.These pleural in-flammatory lesions are often persistent due to pneumonia.The study investigated pleuritis in slaughter pigs in order to determine the prevalence of pleuritis and identify respiratory pathogens involved in pleuro-pneumonia-infected lungs.Two hundreds of lungs from each herd were evaluated using the Slaughterhouse Pleuritis Evaluation System (SPES)from 9 slaughter herds,and the pathogenic bacteria in pleuropneumo-nia-affected lungs were detected.In the present study,52.1% of lungs were found to have different degree of pleuritis.24.0% of the lungs had a SPES score >1,and 26 Actinobacillus pleuropneumonia strains,9 Haemophilus parasuis strains,11 Pasteurella multocida strains,and 15 Streptococcus suis strains were i-dentified in pleuropneumonia-affected lungs.%在屠宰场内,经常可以看到屠宰生猪存在不同程度的胸膜炎病变。这些胸膜上的炎症往往由胸膜肺炎病变持续感染发展而来。试验从宏观角度上统计屠宰生猪出现胸膜炎病变的发生情况,并对胸膜肺炎病变肺脏进行病原菌的分离鉴定。采用屠宰场胸膜炎评估系统(SPES),选取某集约化生猪养殖场的9个畜群,每个畜群随机抽取200份肺脏样品,进行胸膜炎的评价,并随后检测胸膜肺炎的病原菌。其中不同程度的胸膜炎病例占调查肺脏总数的52.1%。打分 SPES>1的病变肺脏比例为24.0%。从胸膜肺炎病变肺脏中分离出胸膜肺炎放线杆菌26株,副猪嗜血杆菌9株,多杀性巴氏杆菌11株和猪链球菌15株。

  6. Establishmen and Application of PCR Method for Detection of Cattle Eperythrozoonosis%肉牛附红细胞体PCR检测方法的建立与初步应用

    Institute of Scientific and Technical Information of China (English)

    许兰娇; 陶清松; 黄志海; 瞿明仁; 何后军; 万根

    2013-01-01

    In order to establish a diagnostic method of molecular biology that can rapidly and accurately detect cattle Eperythrozoon to survey and prevent and control zoonoses, a pair of specific primers were designed according to the published sequence data 16s ribosomal RNA gene of beef cattle Eperythrozoon from Genbank to amplify genomic DNA of cattle Eperythrozoon. The result showed that the targeted gene fragment was 412 bp in length and 98% identical to the published data. The genomic DNA of Escherichia coli, Toxo-plasm and Actinobacillus pleuropneumoniae was amplified by the method, the result was negative. The sensitivity of the assay was 1.23 ng/μL of DNA. It suggested that the method is a rapid, accurate, specific and sensitive diagnostic method for cattle Eperythrozoon.%为建立一个能快速准确的检测出肉牛附红细胞体的分子生物学诊断方法,及时监控好人畜共患病.试验根据Genbank中发表的肉牛附红细胞体16SrRNA序列设计一对特异性引物,通过PCR方法,对肉牛附红细胞体基因组DNA进行扩增.结果表明:扩增出了一段412 bp的DNA片段,与Genbank中发表的肉牛附红细胞体序列同源性为98%.将建立的PCR方法对大肠杆菌、弓形体、胸膜肺炎放线杆菌的DNA进行检测为阴性,检测肉牛附红细胞体DNA的最低浓度为1.23 ng/μL.该方法快速、准确、特异性好、灵敏度高,为肉牛附红细胞体病的临床诊断提供了新的方法.

  7. Occurrence of antimicrobial resistance among bacterial pathogens and indicator bacteria in pigs in different European countries from year 2002 – 2004: the ARBAO-II study

    Directory of Open Access Journals (Sweden)

    Hendriksen Rene S

    2008-06-01

    Full Text Available Abstract Background The project "Antibiotic resistance in bacteria of animal origin – II" (ARBAO-II was funded by the European Union (FAIR5-QLK2-2002-01146 for the period 2003–05. The aim of this project was to establish a program for the continuous monitoring of antimicrobial susceptibility of pathogenic and indicator bacteria from food animals using validated and harmonised methodologies. In this report the first data on the occurrence of antimicrobial resistance among bacteria causing infections in pigs are reported. Methods Susceptibility data from 17,642 isolates of pathogens and indicator bacteria including Actinobacillus pleuropneumoniae, Streptococcus suis and Escherichia coli isolated from pigs were collected from fifteen European countries in 2002–2004. Results Data for A. pleuropneumoniae from infected pigs were submitted from five countries. Most of the isolates from Denmark were susceptible to all drugs tested with the exceptions of a low frequency of resistance to tetracycline and trimethoprim – sulphonamide. Data for S. suis were obtained from six countries. In general, a high level of resistance to tetracycline (48.0 – 92.0% and erythromycin (29.1 – 75.0% was observed in all countries whereas the level of resistance to ciprofloxacin and penicillin differed between the reporting countries. Isolates from England (and Wales, France and The Netherlands were all susceptible to penicillin. In contrast the proportion of strains resistant to ciprofloxacin ranged from 12.6 to 79.0% (2004 and to penicillin from 8.1 – 13.0% (2004 in Poland and Portugal. Data for E. coli from infected and healthy pigs were obtained from eleven countries. The data reveal a high level of resistance to tetracyclines, streptomycin and ampicillin among infected pigs whereas in healthy pigs the frequency of resistance was lower. Conclusion Bacterial resistance to some antimicrobials was frequent with different levels of resistance being observed to

  8. The use of genomic DNA fingerprinting in studies of the epidemiology of bacteria in periodontitis.

    Science.gov (United States)

    Genco, R J; Loos, B G

    1991-07-01

    Recent studies of microbial epidemiology emphasizing the genetic organization and distribution of organisms associated with orofacial infections have led to new insights into the possible origins of pathogenicity. Studies into genetic heterogeneity, acquisition and transmission of these organisms have been markedly advanced by the utilization of the powerful technique of genomic DNA fingerprinting. Characteristic fingerprints for each bacterial isolate can be produced by cleavage of high molecular weight genomic DNA by restriction endonucleases. It is assumed that each DNA fingerprint represents a clonal type. In this report, we review and analyze studies of the epidemiology of bacteria associated with orofacial infections with an emphasis on periodontal disease. Studies of nontypable (NT) Haemophilus influenzae associated with recurrent otitis media illustrate the utility of this technique. DNA fingerprinting clearly demonstrates genetic heterogeneity of NT H. influenzae isolates, and clonality of infection of any individual. Furthermore, DNA fingerprinting has shown that the same clonal type is seen in siblings concurrently suffering from otitis media, suggesting horizontal transmission within the family. Studies of mutans Streptococci also show extensive genetic heterogeneity and show vertical transmission of a predominant clonal type only from mother to infant, but not from father to infant. Studies of Actinobacillus actinomycetemcomitans show considerable genetic heterogeneity among monkey isolates. Thus far, three clonal types have been reported with DNA fingerprinting among isolates from periodontal patients, but additional genetic heterogeneity can be found using specific DNA fragments as probes in hybridization experiments. Intrafamilial transmission of A. actinomycetemcomitans has been demonstrated. Porphyromonas (Bacteroides) gingivalis shows extensive genetic heterogeneity and case reports suggest clonal infection of any one individual. In contrast

  9. Detection of periodontal pathogens in the patients with aortic aneurysm

    Institute of Scientific and Technical Information of China (English)

    Ding Fang; Lyu Yalin; Han Xiao; Zhang Hai; Liu Dongyu; Hei Wei; Liu Yinhua

    2014-01-01

    Background The occurrence and development of aortic aneurysm (AA) are associated with infection.Some researchers have detected the DNA of periodontal pathogens in AA samples in certain populations.However,it has not been done in Chinese population.The objective of this study was to evaluate the prevalence of periodontal pathogens in oral tissue samples and aneurysm samples of AA patients.Methods Eighty-nine subjects with AA and 59 subjects without AA were examined.Periodontal clinical parameters were evaluated.Unstimulated saliva and subgingival plaque somples were collected from all subjects.Twenty-six dissected AA samples were obtained.Evidence of eight periodontal pathogens including Porphyromonas gingivalis (Pg),Actinobacillus actinomycetemcomitans (Aa),Prevotella intermedia (Pi),Tannerella forsythensis (Tf),Treponema denticola (Td),Campylobacter rectus (Cr),Fusobacterium nucleatum (Fn),and Prevotella nigrescens (Pn) was ascertained in all samples by 16S rRNA-based polymerase chain reaction (PCR) assay.Results The periodontal indexes including plaque index (PLI),probing depth (PD),bleeding index (BI),and clinical attachment loss (CAL),of the six Ramfjord index teeth were significantly higher in the AA group than those in the control group (P <0.01).Eight periodontal pathogens in subgingival plaque samples were more frequently detected in the AA group than in control group.The difference in prevalence between the groups was significant for six (out of eight) periodontal pathogens assayed (Pg,Pi,Fn,Pn,Tf,and Td,P <0.01).Additionally,all eight periodontal pathogens were more frequently detected in saliva samples of the AA group than in those of the control group,again with six (out of eight) (Pg,Pi,Fn,Cr,Tf,and Td) displaying significant differences in prevalence between the two groups (P <0.01).Out of 26 aneurysm samples examined,Pg,Pi,Fn,Crand Tfwere detected in 6 (23.1%),2 (7.7%),3 (11.5%),1 (3.8%),2 (7.7%),respectively,and Aa,Pn,and Td were not

  10. Antibacterial Effect of Aloe Vera Gel against Oral Pathogens: An In-vitro Study

    Science.gov (United States)

    Jain, Supreet; Rathod, Nirav; Sur, Jaideep; Laheji, Afshan; Gupta, Naveen; Agrawal, Priyanka; Prasad, Swati

    2016-01-01

    Introduction Natural herbal remedies have shown promising anti-microbial property and fewer side effects compared to synthetic anti-microbial therapy. Aloe Vera is a medicinal plant used for management of various infections since ancient times as it has anti-inflammatory, anti-microbial, and immune-boosting properties. Aim The aim of the present study was to determine the anti-microbial and inhibitory activities of various concentration of Aloe Vera Gel (AVG) against oral pathogenic bacteria. Materials and Methods Subgingival calculus and aspiration of periapical abscess and periodontal abscess was done in 20 patients and the sample transferred to thioglycolate broth, which was incubated in Mutans Sanguis agar, blood agar and cultured in anaerobic gas chamber. The colonies formed were identified further by gram staining methods and biochemical fermentation tests (IMViC). Each isolated colony of identified bacteria were cultured separately in Muller-Hilton broth and incubated at 37°C for 24 hours. Anti-microbial activity of the AVG was tested by the disc diffusion method and minimum inhibitory concentration was determined by broth micro-dilution method. Result Various staining and biochemical tests confirmed that the sample contained Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans), Clostridium bacilli (C. bacilli), Streptcoccus mutans (S. mutans) and Staphlococcus aureus (Staph. aureus). AVG showed anti-bacterial property at 100% and 50% concentration (‘t’ value = 7.504, p-value <0.001). At lower concentration there was no effect against the bacteria. At 100% AVG concentration, zone of inhibition measured was 6.9mm in A. actinomycetemcomitans, 6.3mm in C. bacilli, 6.8mm in S. mutans and 6.6mm in Staph. aureus. The standard drugs were also used to compare anti-bacterial property of AVG. Result showed that higher concentration (100%, 50%) of AVG has comparable zone of inhibition with Ofloxacin (5mcg) and Ciprofloxacin (30mcg). Conclusion AVG at

  11. Cleavage of host cytokeratin-6 by lysine-specific gingipain induces gingival inflammation in periodontitis patients.

    Directory of Open Access Journals (Sweden)

    Salunya Tancharoen

    Full Text Available Lysine-specific gingipain (Kgp is a virulence factor secreted from Porphyromonas gingivalis (P. gingivalis, a major etiological bacterium of periodontal disease. Keratin intermediate filaments maintain the structural integrity of gingival epithelial cells, but are targeted by Kgp to produce a novel cytokeratin 6 fragment (K6F. We investigated the release of K6F and its induction of cytokine secretion.K6F present in the gingival crevicular fluid of periodontal disease patients and in gingipain-treated rat gingival epithelial cell culture supernatants was measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer-based rapid quantitative peptide analysis using BLOTCHIP. K6F in gingival tissues was immunostained, and cytokeratin 6 protein was analyzed by immunofluorescence staining and flow cytometry. Activation of MAPK in gingival epithelial cells was evaluated by immunoblotting. ELISA was used to measure K6F and the cytokines release induced by K6F. Human gingival fibroblast migration was assessed using a Matrigel invasion chamber assay.We identified K6F, corresponding to the C-terminus region of human cytokeratin 6 (amino acids 359-378, in the gingival crevicular fluid of periodontal disease patients and in the supernatant from gingival epithelial cells cultured with Kgp. K6F antigen was distributed from the basal to the spinous epithelial layers in gingivae from periodontal disease patients. Cytokeratin 6 on gingival epithelial cells was degraded by Kgp, but not by Arg-gingipain, P. gingivalis lipopolysaccharide or Actinobacillus actinomycetemcomitans lipopolysaccharide. K6F, but not a scrambled K6F peptide, induced human gingival fibroblast migration and secretion of interleukin (IL-6, IL-8 and monocyte chemoattractant protein-1. These effects of K6F were mediated by activation of p38 MAPK and Jun N-terminal kinase, but not p42/44 MAPK or p-Akt.Kgp degrades gingival epithelial cell cytokeratin 6 to K6F that, on

  12. Antimicrobial Constituents of Artemisia afra Jacq. ex Willd. against Periodontal Pathogens

    Directory of Open Access Journals (Sweden)

    Garland More

    2012-01-01

    Full Text Available The phytochemical investigation of an ethanol extract of Artemisia afra led to the isolation of six known compounds, acacetin (1, 12α,4α-dihydroxybishopsolicepolide (2, scopoletin (3, α-amyrin (4, phytol (5, and a pentacyclic triterpenoid betulinic acid (6. The compounds were evaluated for antimicrobial activity against Gram positive (Actinomyces naeslundii, Actinomyces israelii, and Streptococcus mutans, Gram negative bacteria (Prevotella intermedia, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans previously known as Actinobacillus actinomycetemcomitans, and Candida albicans. The crude extract of A. afra inhibited the growth of all tested microbial species at concentration range of 1.6 mg/mL to 25 mg/mL. The compounds 1–6 also showed activity range at 1.0 mg/mL to 0.25 mg/mL. Three best compounds (scopoletin, betulinic acid, and acacetin which showed good antimicrobial activity were selected for further studies. Cytotoxicity of extract and compounds was determined using the XTT cell proliferation kit. The antioxidant activity of the extract and compounds was done using the DPPH scavenging method. The extract showed good antioxidant activity with an IC50 value of 22.2 μg/mL. Scopoletin had a strong transformation of the DPPH radical into its reduced form, with an IC50 value of 1.24 μg/mL which was significant to that of vitamin C (1.22 μg/mL. Acacetin and betulinic acid exhibited a decreased scavenging activity with the IC50 of 2.39 and 2.42 μg/mL, respectively. The extract and compounds showed moderate toxicity on McCoy fibroblast cell line and scopoletin was relatively nontoxic with an IC50 value of 132.5 μg/mL. Acacetin and betulinic acid also showed a smooth trend of non-toxic effects with IC50 values of 35.44 and 30.96 μg/mL. The obtained results in this study confirm the use of A. afra in the treatment of microbial infections.

  13. 慢性牙周炎患者的龈下微生物与吸烟状况%Subgingival microflora and smoking status in chronic periodontitis

    Institute of Scientific and Technical Information of China (English)

    徐莉; B·G·Loos; 等

    2002-01-01

    目的评价慢性牙周炎患者的吸烟状况与龈下牙周致病微生物的百分比.方法 112例慢性牙周炎患者,根据吸烟状况分为:①重度吸烟组>10支/天(n=32);②轻度吸烟组≤10支/天(n=18);③戒烟者组(n=24);④非吸烟组(n=38).观察者口内每个象限,选取探诊最深的1或2个位点, 纸捻法取龈下菌斑,厌氧培养;并测量该位点的探诊深度(Probe Depth, PD)、附着丧失(Attachment Loss,AL)和探诊后出血(Bleeding of probe,BOP).结果①取样位点临床指标的均值为PD:6.3mm、AL:6.5mm及BOP:89%,各组间无显著性差异.②组间菌落形成单位和龈下微生物伴放线放线杆菌(Actinobacillus actinomycetemcomitans,A.a)、牙龈卟啉单胞菌(Porphyromanas gingivalis,P.g)、中间普氏菌(Prevotella intermedia,P.i)、核梭杆菌(Fusobacterium nucleatum,F.n) 和微消化球菌(Peptostreptococcus micros,P.m)的百分比均无差异.③方差分析显示仅轻度吸烟组福赛类杆菌(Bacteroides forsythus,B.f)的百分比略高于其它组(P<0.04).结论慢性牙周炎患者,探诊深度和附着丧失相似的位点,吸烟组、戒烟组和非吸烟组的龈下牙周致病微生物百分比(B.f除外)无显著差异.

  14. A taxonomic study of the genus Haemophilus, with the proposal of a new species.

    Science.gov (United States)

    Kilian, M

    1976-03-01

    is uncertain and its possible relationship to Actinobacillus actinomycetemcomitans requires further study. The positive correlation found between the ecology of the strains studied and their affiliation with the different taxa is discussed.

  15. 中华慈姑中具有抗菌活性的对映-玫瑰烷和对映-贝壳杉烷二萜%Antibacterial ent-Rosane and ent-Kaurane Diterpenoids from Sagittaria trifolia var. sinensis

    Institute of Scientific and Technical Information of China (English)

    刘雪婷; 施瑶; 梁敬钰; 闵知大

    2009-01-01

    AIM: To investigate the antibacterial components from Sagittaria trifolia vat. sinensis. METHODS: The separation was performed over silica gel and Sephadex LH-20 column. The structures of the obtained compounds were determined on the basis of NMR data and chemical methods. RESULTS: Ten diterpenoids were isolated. Sagittine H (1), sclareol (2), and 19-β-L-3'-acetoxyarabinofuranosyl- ent-kaur-16-ene-19-oate (3) were evaluated for their abilities to inhibit the growth of three oral pathogens, Streptococcus mutans ATCC 25175, Actinomyces naeslundiis ATCC 12104, and Actinobacillus actinomycetemeomitans ATCC 43717, using minimum inhibitory concentration (MIC) assays. CONCLUSIONS: sagittine H (1) is a new ent-rosane glycoside and demonstrated MIC of 62.5 μg·mL-1 against both S. mutans and A. naeslundiis. Compound 3 exhibits strong activity against S. mutans and A. naeslundiis with MIC of 15.6 μg·mL-1 for both.%目的:研究中华慈姑中的抗菌活性成分.方法:通过各种硅胶柱层析和凝胶柱层析进行分离和纯化.分离得到的化合物通过一维和二维的核磁技术以及化学方法进行结构解析.结果:从中华慈姑中分离得到10个化合物,并测试了化合物1-3抗3种口腔细菌的活性.结论:化合物1是一个新的对映-玫瑰烷二萜苷,化合物3显示了很强的抗两种口腔细菌(Streptococus mutans和Actinomyces naeslundiis)的活性,MIC均为15.6 μg·mL-1.化合物1的活性较弱,对上述两种口腔细菌的MIC均为62.5μg-mL-1.

  16. 1株鸽源鸡杆菌复合群3的分离和鉴定%Isolation and identification of one pigeon-origin Gallibacterium genomo sp3

    Institute of Scientific and Technical Information of China (English)

    杨晓林; 陈英; 余松城; 邹年莉; 吴瑞婷; 夏静; 牛婷; 王富妍; 黄勇

    2014-01-01

    .This disease had caused great economic losses to the pigeon farm.In order to propose a proper treatment program for this disease,representative samples were collected from disease pigeons for bacteria isolation and virus detection. The bacteria were isolated by blood agar and were identified by following test:morphological observation, biochemical identification,animal experiment,antibiotic sensitive experiment,1 6S rRNA gene sequence analysis. Animal experiments were conducted by infecting mice and pigeons to observe its pathogenicity.For the virus detection,the nucleotide sequences of infectious bronchitis virus (IBV),avian influenza virus subtype (H 9 N2) and Newcastle disease virus (NDV) were detected by RT-PCR. The results showed that NDV was detected from the lung and trachea of some sick pigeons,and one bacterial strain was isolated from the livers and lungs of sick pigeons and named as SC01.The SC01 strain was gram-negative (G-),short rod-shaped bacteria.It was pathogenic to mice,and the median lethal dose to mice was 1.26 × 10 9 CFU/mL.Biochemical test showed that it can ferment most sugars and alcohols.Sensitive drug screen revealed that the SC01 strain was sensitive to ceftriaxone sodium,cefoperazone sodium,florfenicol,cephalexin,cefazolin,and resistant to neomycin,streptomycin,erythromycin,sulfafurazole,etc.The 1 6S rRNA gene sequence of the SC01 strain was blasted in GenBank,and the results showed that the nucleotide sequence homologies of SC01 and Gallibacterium ,Pasteurella , Haemophilus ,Actinobacillus were relatively high. The 1 6S rRNA nucleotide sequence homologies of SC01 and four G.genomo sp.3 strains were 98.5% 99.5%,and the highest homology (99.5%) came from the comparison between the sequences of SC01 and EU423996.Phylogenetic analysis showed that SC01 strain and nine Gallibacterium strains formed an independent big branch in phylogenetic tree.The SC01 strain and G. genomo sp. 3 fell into the same small branch. Meanwhile, Pasteurella , Haemophilus and

  17. 长春市某小学7~12岁儿童牙周致病菌分布状态调查%Distribution of periodontal pathogens in dental plaque samples from 7 to 12-years-old children of Changchun Ziqiang primary school

    Institute of Scientific and Technical Information of China (English)

    倪雪岩; 伊田博; 鈴木基之; 吕亚林; 魏秀峰; 井上美津子

    2010-01-01

    目的 应用聚合酶链反应(PCR)法对儿童口腔内牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)分布状态进行检测,探讨检出结果与牙周临床指标之间的关系.方法 选取长春市自强小学151名7至12岁儿童为研究对象,选择右上颌中切牙唇面和右上颌第一磨牙颊面为被检部位,取龈上菌斑、记录探诊出血(bleeding on probing,BOP)、探诊深度(probing depth,PD)、牙龈指数(gingival index,GI),应用PCR法对两菌种进行检测.结果 ①儿童龈上菌斑中Pg、Aa检出率为27.6%、54.3%;②6颊面Pg、Aa的检出率(40.0%、57.9%)均高于1 唇面(15.5%、50.7%),Pg检出率差异有统计学意义(P<0.01),且与BOP、PD、GI呈正相关;③Pg检出率随年龄增长呈逐渐增高趋势,Aa检出率在11~12岁组最高,其次为7~8岁组和9~10岁组;④BOP阳性部位Pg、Aa检出率(38.3%、65.4%)均高于BOP阴性部位(23.2%、50.5%),P<0.05.在BOP阳性部位,随PD加深Pg检出率逐渐增高,特别是在PD≥4mm时,Pg检出率明显增高(P<0.05),显示Pg检出率与BOP阳性、PD增加呈正相关.结论 7~12岁儿童龈上菌斑中高频度分布着Pg、Aa;上颌前牙区与磨牙区菌丛构成不同,Pg在磨牙区定植更早;两菌种检出率随年龄增长而增加,且与牙周临床指标密切相关,儿童早期采取牙周病的预防措施是非常必要的.%Objective To detect the presence of Porphyromonas gingivalis (Pg) and Actinobacillus actinomycetemcomitans(Aa) using polymerase chain reaction (PCR) in the oral plaque samples from children and investigate the relationship between bacteria and clinical parameters. Methods A total of 151 children aged 7 to 12 years were selected from Changchun Ziqiang primary schooL The supragingival plaque sample was collected from the mesiobuccal and labial surfaces of the right maxillary central incisor and the right maxillary first molar. Extracted DNA from plaque

  18. Detection of putative periodontal pathogens in the subgingival plaque of type Ⅱ prediabetes children with gingivitis%Ⅱ型糖尿病前期伴牙龈炎患儿牙周可疑致病菌的检测

    Institute of Scientific and Technical Information of China (English)

    蔡梁婧; 邰佰霞; 王永兰; 郭宏磊; 朱志颖

    2012-01-01

    目的:检测Ⅱ型糖尿病前期伴牙龈炎患儿龈下菌斑中的牙周可疑致病菌,指导牙周病的治疗和预防.方法:以6~14岁的儿童作为研究对象,按照血糖水平和牙龈炎症状况分为3组:糖尿病前期伴牙龈炎组(实验组)、代谢正常伴牙龈炎组(对照组一)、代谢正常而牙周健康组(对照组二),每组各30例.以牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)、具核梭杆菌(Fusobacterium nucleatum,Fn),中间普氏菌(Prevotella intermedia,Pi),福塞坦氏菌(Tannerella forsythia,Tf)为目标菌,应用以16SrRNA为基础的聚合酶链反应(polynerase chain reaction,PCR)技术对龈下菌群进行检测.分析5种菌的相对含量与牙周临床指标和血糖水平的相关性.结果:5种可疑致病菌的检出率实验组明显高于对照组二(P<0.05);实验组虽高于对照组一,但无统计学差异(P>0.05).Pg的相对含量与牙周探诊深度(Probing depth,PD)、龈沟出血指数(Sulcus bleeding index,SBI)以及空腹血糖(Fasting blood-glucose,FBG)存在相关性(P<0.05);Pi的相对含量与PD、SBI、FPG、口服葡萄糖耐量实验2h血糖(OGTT2hBG)存在相关性(P<0.05);Aa和Fn的相对含量与SBI、PD存在相关性(P<0.05);Tf的相对含量与SBI、PD、FBG存在相关性(P<0.05).结论:Ⅱ型糖尿病前期伴牙龈炎者牙周可疑致病菌的检出率明显高于正常对照组,虽与单纯牙龈炎组相比,其数量无明显差异,但应更加注意血糖水平和菌斑控制.%AIM; To detect putative periodontal pathogens in the subgingival plaque of type II prediabetes children with gingivitis. METHODS : Six to 14 years old type II prediabetes children with gingivitis (experimental group) , children with only gingivitis ( control) and healthy children ( normal control) were included in this study, with 30 subjects in each group. Subgingival plaque was taken for the detection of

  19. Periodontopathogens in the saliva and subgingival dental plaque of a group of mothers Periodontopatógenos na saliva e placa subgengival de um grupo de mães

    Directory of Open Access Journals (Sweden)

    Odila Pereira da Silva Rosa

    2002-12-01

    Full Text Available The aim of this study was to assess the periodontal condition and the presence of putative periodontal pathogens in 30 Brazilian mothers, aging 21-40 years (28.4 ± 4.49 years, and in their children, aging 5-6 years, since mothers can be a source of pathogens and, thus, influence their children's bacteriological and clinical condition. Besides assessing the plaque index (PI, gingival index (GI and pocket probing depth (PD, the survey analyzed four subgingival dental plaque samples from mothers and children, as well as a sample of stimulated saliva from mothers. Those samples were analyzed by means of the slot immunoblot (SIB technique, in order to determine the presence of Actinobacillus actinomycetemcomitans (Aa, Prevotella nigrescens (Pn, Porphyromonas gingivalis (Pg and Treponema denticola (Td. The mean values and standard deviations of the evaluated clinical variables for mothers and children were, respectively: 1.86 ± 0.67 and 1.64 ± 0.68 for PI, and 1.24 ± 0.67 and 0.82 ± 0.37, for GI. Only for mothers, the total PD was 1.81 ± 0.69 mm, and the PD of four sites was 4.03 ± 1.40 mm. The Wilcoxon test revealed significant difference (p Procurou-se avaliar a condição periodontal e a presença de periodontopatógenos em 30 mães brasileiras, com idades entre 21-40 anos (28,4 ± 4,49 anos e seus filhos, com 5-6 anos de idade, considerando que elas possam ser fonte de transmissão para seus filhos e influenciar suas condições clínicas e bacteriológicas. Além de determinar o índice de placa (IP, índice gengival (IG de mães e filhos, e a profundidade de sondagem periodontal (PS, apenas das mães, avaliaram-se quatro amostras de placa dental subgengival de mães e filhos e uma amostra de saliva total estimulada das mães para a presença de Actinobacillus actinomycetemcomitans (Aa, Prevotella nigrescens (Pn, Porphyromonas gingivalis (Pg e Treponema denticola (Td, pela técnica de "slot immunoblot" (SIB. As médias e desvios

  20. 模拟高原缺氧环境对家兔慢性牙周炎龈下菌群种类分布的影响%Distribution of subgingival microbial category in periodontitis rabbits under simulated high altitude hypoxia condition

    Institute of Scientific and Technical Information of China (English)

    罗丽; 张纲; 祝金香; 谭颖徽; 高钰琪

    2012-01-01

    目的 模拟高原缺氧环境,构建兔牙周炎动物模型,研究其龈下菌斑中的主要牙周致病菌与平原组的差异.方法 选取清洁级家兔40只完全随机设计分成4组:平原实验组、平原对照组、高原实验组、高原对照组各10只,实验组采用正畸结扎丝结扎双下颌前牙,并给予高糖饮食;高原组置模拟海拔5000m的低压氧舱,每天缺氧23 h,持续饲养8周.8周后提取龈下菌斑基因组DNA,用PCR扩增细菌16S rDNA片段方法检测标本中6种牙周炎致病菌包括牙龈卟啉单胞菌(Dorhyromonas gingivalis,Pg)、福赛斯坦纳菌(Bacterides forsythus,Bf)、伴放线放线杆菌(Actinobacillus actinomycetemcomitans,Aa)、中间普氏菌(Prevotella intermedia,Pi)、具核梭杆菌(Fusobacterium nucleatum,Fn)和齿垢密螺旋体(Trepone ma denticola,Td)的检出率.结果 与平原实验组相比,高原实验组的牙龈出血指数、菌斑指数和牙周袋深度均有统计学差异(P<0.05).牙周可疑致病菌Pg、Bf Aa、Pi、Fn和Td在高原实验组和平原实验组都可检出,其中高原实验组Fn的检出率80%高于平原实验组20%(P<0.05);平原实验组Pi的检出率70%高于高原实验组15% (P <0.05).结论 高原低氧环境是牙周炎加重的主要原因,Fn与高原牙周炎病变程度加重有一定关系.%Objective To investigate the difference of main pathogenic bacteria in subgingival plaque of periodontitis rabbits under normal oxygen and simulated high altitude hypoxia conditions. Methods A total of 40 rabbits were randomly divided into 4 groups including a normal oxygen periodontitis group, a normal control group, a hypoxia periodontitis group and a hypoxia control group (n = 10). Rabbits of the periodontitis groups were given ligation of anterior teeth by steel wire and high-carbohydrate diet, while rabbits of the hypoxia groups were raised in a low pressure oxygen cabin simulating high altitude hypoxia condition (5 000 m) , 23 h

  1. Endocardite infecciosa por Haemophilus aphrophilus: relato de caso Infective endocarditis due to Haemophilus aphrophilus: a case report

    Directory of Open Access Journals (Sweden)

    Ricardo M. Pereira

    2008-04-01

    Full Text Available OBJETIVO: Descrever o caso de uma criança com endocardite infecciosa causada por Haemophilus aphrophilus. DESCRIÇÃO: Menino com febre e calafrios há 20 dias. À internação, apresentava-se febril, descorado e sem sinais de instabilidade hemodinâmica; à ausculta cardíaca, tinha sopro holosistólico em foco mitral. Os exames laboratoriais identificaram anemia (hemoglobina = 9,14 g/dL, leucócitos totais de 11.920 mm³, plaquetas de 250.000 mm³, velocidade de sedimentação das hemácias e proteína C reativa elevadas. O ecocardiograma revelou imagem em válvula mitral, sugestiva de vegetação. Com a hipótese de endocardite, foi iniciada antibioticoterapia com penicilina cristalina (200.000 UI/kg/dia associada à gentamicina (4 mg/kg/dia. No terceiro dia de tratamento, foi identificado Haemophilus aphrophilus em hemoculturas, sendo então trocado o esquema antibiótico para ceftriaxona (100 mg/kg/dia. No 20º dia de internação, encontrava-se pálido, mas sem febre e sem outras queixas. Os exames mostravam hemoglobina = 7,0 g/dL, leucócitos = 2.190 mm³, plaquetas = 98.000 mm³, razão normatizada internacional = 1,95 e R = 1,89. Foi feita hipótese de reação adversa ao ceftriaxona, que foi substituído por ciprofloxacina, 20 mg/kg/dia, até completar 6 semanas de tratamento. Após 72 horas da troca, houve normalização dos exames. Durante seguimento ambulatorial, apresentou insuficiência mitral grave, sendo submetido a troca de válvula por prótese metálica 9 meses após quadro agudo. Há 3 anos encontra-se bem, em acompanhamento ambulatorial. COMENTÁRIOS: É rara a identificação de agentes do grupo HACEK (Haemophilus ssp, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens e Kingella kingae em crianças com endocardite infecciosa. O caso apresentado, sem fatores de risco relacionados a esses agentes, reafirma a necessidade de tentar sempre identificar o agente etiológico das endocardites para

  2. 牙周致病菌密度感应信号系统luxS基因的检测%Detection of luxS gene of quorum sensing signal system in periodontal pathogen

    Institute of Scientific and Technical Information of China (English)

    雷朝锋; 杨禾; 孙昌娟; 苗棣; 徐屹

    2009-01-01

    目的 检测牙周可疑致病菌密度感应信号系统luxS基因,了解其在牙周致病菌中的分布.方法 选取牙龈卟啉单胞菌、伴放线放线杆菌、具核梭杆菌的模式株、参考株及临床分离株作为研究对象,提取DNA,通过聚合酶链反应(PCR)、电泳鉴定和DNA测序,并利用GenBank数据库的Blast检测以上细菌luxS基因的存在情况.结果 电泳鉴定存在目的 条带,测序和Blast检测表明牙龈卟啉单胞菌PCR产物与目的 基因有高度一致性(均为99%以上),具核梭杆菌测序结果 与GenBank数据库的基因相同,伴放线放线杆菌电泳鉴定结果 显示存在目的 条带(750 bp),与参考条带大小一致.结论 本实验引物设计合理,能较好地扩增出牙龈卟啉单胞菌、具核梭杆菌、伴放线放线杆菌各实验菌株的luxS基因,为进一步研究luxS基因的功能奠定了基础.%Objective To detect the presence and distribution of luxS gene in quorum sensing signal system in the periodontal pathogens.Methods The total DNA of Porphyromonas gingivalis(Pg),Fusobacterium nucleatum(Fn),Actinobacillus acitinomycetimcomtans(Aa)were extracted.The presence of luxS was detected by polymerase chain reaction(PCR).The products of PCR were detected by electrophoresis.sequenced and identified by a Blast search of the GenBank database. Results Electrophoresis,sequencing and Blast searching indicated that the PCR products of Pg were highly consistent with the luxS gene in GenBank.The sequencing result of Fn was also identified with the target gene.The PCR product of Aa was the same as reference through electrophoresis.Conclusions Pg,Fn,Aa contain luxS gene.Further studies may be required to investigate the functions of luxS in the periodontal pathogens.

  3. 山西省输变电工程水土保持低扰动工程技术%Low Disturbance Engineering Technology of Soil and Water Conservation in Power Transfer and Transformation Construction Project in Shanxi Province

    Institute of Scientific and Technical Information of China (English)

    王露露; 孙中峰; 朱清科

    2013-01-01

    Power transfer and transformation projects have the large span.So their natural conditions,social conditions,economic conditions and the source of soil and water loss are complex.Moreover,the projects temporarily cover large area and disturb a wide range.To explore the low disturbance engineering technology in soil and water conservation of power transmission construction and to help optimize the design of both the main work and soil and water conservation measures,eighteen 500 kV power transfer and transformation projects in Shanxi Province were selected as research samples.By comprising the extent of disturbances and the amount of soil and water losses caused by the traditional main work and soil and water conservation engineering and the low disturbance ones,it was found that the low disturbance engineering technology,including unequal legs and basis transmission tower,haulage methods of aerial cableway and actinobacillus without touching the ground,can significantly reduce the ranges of disturbance and the amount of soil and water loss.In the design of soiland water conservation measures,if the suitable surface soil stripping technology can be used based on the feature of soil and water loss and the soil condition in different control areas and temporary covering technology can be used in some temporary land,the secondary disturbance as well as the investments of the soil and water conservation engineering can be reduced.It was suggested that the low disturbance engineering technology which can reduce soil and water loss should be given priority in the design of main works as well as soil and water conservation measures of power transfer and transformation projects.%输变电工程跨度大,自然、社会经济条件及水土流失来源复杂多变,且临时占地多,扰动范围大.为探索输变电工程水土保持低扰动工程技术,为输变电主体工程及水土保持措施优化设计提供参考依据,以山西省18项500 kV输变电工程为研

  4. Antibacterial activity of synthetic antimicrobial decapeptide against oral bacteria%人工合成抗菌肽对口腔细菌抗菌性能的初步研究

    Institute of Scientific and Technical Information of China (English)

    刘奕; 费伟; 王丽娜; 董广艳; 吴红崑

    2014-01-01

    Objective This study aims to evaluate the antimicrobial activity of decapeptide, a novel antimicrobial peptide, against several major cariogenic and periodontopathogenic bacteria in vitro. Methods In this study, we investigated the antimicrobial activity of decapeptide against Streptococcus mutans (S.mutans), Streptococcus sobrinus, Lactobacillus acidophilus, Streptococcus sanguis, Streptococcus gordonii, Actinomyces viscosus, Actinomyces naeslundii, Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, and Saccharomyces albicans in vitro using the agar diffusion method and broth dilution method. Furthermore, a time–kill kinetic study of decapeptide against S. mutans was performed. Results The results showed that decapeptide exhibited antimicrobial activity against various oral bacteria and fungi. The minimum inhibitory concentration (MIC) of main cariogenic bacteria ranged from 62.5 μg·mL-1 to 125 μg·mL-1, and the MIC of periodontopathogenic bacteria tested ranged from 250 μg·mL-1 to 1 000 μg·mL-1. Among the bacteria tested, decapeptide had a strong inhibitory effect on cariogenic S.mutans. Results of the time–kill kinetic studies showed that decapeptide reduced the viable counts of S. mutans by more than one order of magnitude after 20 min of incubation, and thoroughly killed S.mutans after 30 min. No viable cells could be detected after 24 h of incubation. Conclusion This study suggest that decapeptide might have potential clinical application in treating dental caries by killing S.mutans within dental plaque.%目的:  评价人工合成抗菌肽(十肽)对口腔常见感染性疾病主要致病菌的抑菌活性。方法  采用琼脂扩散法及液体稀释法体外评价十肽对变异链球菌、表兄链球菌、嗜酸乳杆菌、血链球菌、格氏链球菌、黏性放线菌、内氏放线菌、牙龈卟啉单胞菌、中间普雷沃菌、具核梭杆菌、伴放线嗜

  5. Effect of periodontal surgery on the subgingival periodontal microbiota in artificial class Ⅲ furcation defects: an experimental study in monkeys%牙周手术治疗对猴Ⅲ度根分叉病变模型龈下菌斑中牙周致病微生物的影响

    Institute of Scientific and Technical Information of China (English)

    朱卫东; 侯建霞; 刘凯宁; 唐晓琳; 孟焕新

    2012-01-01

    Objective: This study was to longitudinally evaluate the change of prevalence of five perio-dontal putative pathogens in the subgingival plaque of artificial class IE furcation defects at the three time-points , including before the establishment of furcation defects, before and 6 months after periodontal surgery. Methods; Eighteen chronic infected class Ⅲ FI defects were created at the mandibular first molars , second molars and second premolars of three adult male Macaca fascicularis. The samples of subgingival plaque were obtained from the subgingival area of furcation defects in buccal and lingual sites before the establishment of furcation defects, before and 6 months after periodontal surgery. 36 samples were obtained at one time-points. Five periodontal putative pathogens, including Porphyromonas gingivalis (Pg), Tannerella forsythensis (Tf), Treponema dinticola (Td), Actinobacillus actinomycetemcomitans (Aa) and Fusobacterium nucleatum (Fn) , were detected with 16SrRNA based PCR. Results; 1. The prevalence of Pg, Tf, Td and Fn was gradually increased, from 58. 3% to 69.4% to 88. 9% , 47,2% to 69.4% to 83. 3%, 13.9% to 36. 1% to 61.1% (P<0.01), and 69. 4% to 91. 7% to91.7%(P<0.05) , respectively during the experimental period. The prevalence of Fn was higher than Pg, Tf and Td. The prevalence of Aa was the lowest and no obvious difference among the three samplings ( from 25. 9% to 13. 9% to 33. 3% ) was detected. 2. The prevalence of more than 3 species simultaneously detected was increased from 38. 9% to61.1% to 83.3% (P <0.01). The red complex (Pg + Tf + Td) was detected from 8. 3% to 27. 8% to 44.4% (P <0. 01) at the different time point. 3. The combined detection frequency of red complex in the inflammatory sites (87. 5% ) , which were histologically defined as inflammatory cells infiltrated in furcation area 6 months post-surgery, and the same sites pre-surgery (62. 5% ) was more than that in pre-creation of furcation defects (P <0. 01). But there

  6. Effects of smoking on gingival sulcus of pathogenic microorganisms in postoperative patients with chronic periodontitis%吸烟对慢性牙周炎患者术后龈沟内致病微生物的影响

    Institute of Scientific and Technical Information of China (English)

    李泽慧; 姜涵; 徐隽; 李刚

    2015-01-01

    Objective To investigate the effects of smoking on gingival sulcus of pathogenic microorganisms in postoperative patients with chronic periodontitis. Methods According to the smoking status, 60 cases in our hospital from January 2012 to December 2012 with chronic periodontitis were divided into smoking group (n=30) and non- smoking group (n=30).Both groups were treated with modified Widman flap surgery. The following items were set on before operation and 4,8 and 12 weeks after operation, respectively. ①The microorganisms on gingival sulcus such as Actinobacillus actinomycetemcomitans (Aa),Porphyromonas gingivalis (Pg),Forsyth Tanner Rand (Tf), Prevotella intermedia (Pi),Fusobacterium nucleatum (Fn) and Prevotella nigrescens (Pn) were detected by polymerase chain reaction (PCR). ②The clinical indicators of plaque index (PLI),sulcus bleeding index (SBI),probing pocket depth (PPD) were detected.The variations of microbial and clinical index on gingival sulcus were observe and recorded. Results The numerical of PLI, SBI, PPD of two groups were reduced compared before and after operation.The numerical of PLI,SBI,PPD of two groups were reduced significantly in 4 weeks,and the differences were statistically significant ( <0.01).PLI,SBI,PPD of two groups were increased comparison of 8 and 12 weeks after operation with 4 weeks after operation. In the treatment of 4,8 and 12 weeks after operation, the degree of changes in PLI, SBI and PPD were compared between smoking group and non - smoking,the differences (especially two weeks after operation) were statistically significant ( <0.05).Pg,Tf and Pi were the pathogen and they had higher detection rate compared to Aa,Fn and Pn.The site detection rates of Aa,Pg,Tf,Pi,Fn and Pn in 4 weeks after treatment were significantly lower than that before treatment, and the differences were statistically significant ( <0.05).The site detection rates of Pg,Tf and Pi in 8 and 12 weeks after treatment were significantly lower than that

  7. 致农民肺嗜热吸水链霉菌cDNA文库的构建及体外表达%Construction of cDNA library and antigen expression of Streptomyces Thermohydroscopicus in vitro

    Institute of Scientific and Technical Information of China (English)

    凌媛; 王玲玲; 刘朔; 马列; 王群; 张丽娇; 何晓雨; 赵明静; 王笑歌

    2013-01-01

    positive clones with the average length of 495 bp which contained 347 unique genes.The homology analysis revealed that 2 potential antigen-specific genes were highly allied with outer membrane lipoprotein (51 %) and transferring-binding protein B (42%) from Actinobacillus pleuropneumoniae serotype (APP).The molecular weights of the two expression products were 63kDa and 30kDa.Conclusions The cDNA library of Streptomyces Thermohydroscopicus harbors all EST sequences and most likely contains the majority of the virulence genes.It not only provides the basis for elucidating the etiology of farm's lung disease,but also facilitates the development of serological diagnosis and vaccine for farm's lung disease.

  8. Distribution of pathogens causing infections in elderly patients with periodontal disease and related factors%老年牙周病患者感染病原菌的分布与相关因素分析

    Institute of Scientific and Technical Information of China (English)

    彭澜; 刘中林; 王晖; 谢红帼; 严加林

    2016-01-01

    OBJECTIVE To explore the distribution of pathogens causing infections in elderly patients with perio-dontal disease ,analyze the related factors ,and observe the effect of use of antibiotics .METHODS A total of 72 elderly patients with periodontal disease who were treated in the hospital from Jan 2013 to Jan 2014 were recruited as the study objects and assigned as the observation group ,then the pathogens were detected ,the patients were treated with antibiotics on the basis of the detection results ,and the therapeutic effect was observed .Meanwhile , 96 healthy elderly people who received physical examination were chosen as the control group .The baseline data of the two groups of patients were investigated ,the distribution of the pathogens causing the infections was ob-served ,and the related factors for the infections were analyzed .RESULTS There was no significant difference in the age between the observation group and the control group;there was significant difference in the oral hygiene habit ,complication with more than two kinds of underlying diseases ,smoking ,education level ,negative emotion , or complication of diabetic mellitus between the two groups (P<0 .05) .Totally 111 strains of pathogens were iso-lated from 72 patients ,of which 25 .2% were Actinobacillus actinomy ,19 .8% were Porphyromonas gingivalis , and 15 .3% were Capnocytophaga sputigena .The clearance rate of pathogens was 91 .9% by tinidazole ,94 .6%by ornidazole , higher than 70 .3% by metronidazole , and there was significant difference ( P < 0 .05 ) . CONCLUSION There are a variety of factors for the infections in the elderly patients with periodontal disease .It is an effective way to develop good health habit ,regularly carry out the periodontal examination ,and treat the perio-dontal disease in a timely manner so as to avoid the loss of teeth and ensure the oral health .%目的:探讨老年牙周病患者感染病原菌的相关因素及其抗菌药物

  9. Analysis of relevant factors causing periodontal infections to elderly patients and use of antibiotics%老年患者牙周感染病原菌的相关因素分析及抗菌药物应用

    Institute of Scientific and Technical Information of China (English)

    俞丹; 陈舜岳; 王姝; 卢宇

    2014-01-01

    OBJECTIVE To analyze the relevant factors causing periodontal infections to elderly patients and to evaluate the therapeutic effect of antibiotics .METHODS Totally 69 cases of elderly outpatients with periodontal disease from Mar 2012 to Mar 2013 were selected for pathogen detection and were included into study group ;at the same time ,100 cases of old patients examined to be healthy at the same period were included into control group;the basic data of patients were investigated by the self-designed questionnaire and after giving symptomatic antimicrobial treatment ,the therapeutic effects were evaluated .RESULTS Poor oral hygiene habits ,low education levels ,smoking ,2 or more complicated underlying diseases ,complicated diabetes ,and negative emotions were the main factors for the elderly periodontal disease patients suffering pathogen infection ,and there was significant difference (P<0 .05) .Among the 69 cases of infected patients ,a total of 102 strains of anaerobic bacteria were isolated ,and the top three were Actinobacillus actinomycetes (26 .47% ) ,Porphyromonas gingivalis (20 .59% ) and Capnocytophaga sputigena (14 .71% );the patients were given metronidazole ,tinidazole and ornidazole for treatment with the pathogen clearance rates of 78 .26% , 91 .30% and 95 .65% respectively ;the pathogen clearance rates of tinidazole and ornidazole were higher than that of metronidazole ,and there was significant difference between the groups (P< 0 .05) .CONCLUSIONS The incidence of periodontal disease in the elderly patients is high and there are many factors causing pathogen infection ,so it is required to go for periodontal examinations at regular intervals ,pay more attention to oral hygiene and to timely treat periodontal diseases ,which are effective ways to maintain oral health and avoid tooth loss .%目的:对老年患者牙周感染病原菌的相关因素进行分析,并对抗菌药物治疗效果进行评价。方法选择2012年3月-2013年3

  10. Is periodontal disease a public health issue in Colombia?

    Directory of Open Access Journals (Sweden)

    Jorge Hernán Ramírez

    2007-09-01

    Full Text Available Oral health inequalities are a good indicator of socioeconomic contrasts in a country. This could be explained because people living in low socioeconomic strata receive an insufficient and inadequate education in oral health, lack the economic resources to visit a dentist, and frequently cannot acquire basic oral hygiene products. Furthermore, the limited economic resources designated in public health programs for oral diseases prevention and treatment are based in the misconception of separating oral health from general health and the idea of dental treatment as «esthetic» without recognizing the negative impact on people quality of life secondary to dental pain and tooth loss1. In 2003, the World Health Organization (WHO emphasized the need to promote oral health and published a guidance document for every nation to define their goals in oral health indicators by the year 20202. Periodontal disease is highly prevalent worldwide and is also a major cause of tooth loss. Gingivitis, the mildest form of periodontal disease is caused by the biofilm (bacterial plaque accumulating in the tooth surface adjacent to the gingival tissue (gums. Periodontitis, the more severe type of periodontal disease is characterized by the destruction of connective tissue and dental bone support after an inflammatory host response secondary to infection by periodontal bacteria (Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Treponema denticola, Tannarella forsythensis, among others3. Untreated periodontitis may eventually cause tooth loss. In accordance with the Third National Survey of Oral Health (ENSAB III 50.2% of people living in Colombia have some degree of periodontal disease. Among those, 17.7% of the cases are classified from moderate to severe periodontitis4. Periodontitis causes an inflammatory response with elevation of multiple acute phase reactants like fibrinogen and C-Reactive Protein (CRP5. In diabetic patients periodontitis is

  11. ¿Se debe considerar a la enfermedad periodontal un problema de salud pública en Colombia?

    Directory of Open Access Journals (Sweden)

    Jorge Hernán Ramírez

    2007-09-01

    Full Text Available Las desigualdades en salud oral se han considerado como un fiel reflejo de los contrastes socioeconómicos de un país. Esto se explica porque los estratos más bajos reciben una inadecuada e insuficiente educación en el cuidado de la salud oral, carecen de los recursos suficientes para acceder a una consulta odontológica y con frecuencia no pueden adquirir los artículos básicos de higiene oral. Además, los escasos recursos asignados en programas de salud pública para la prevención y el tratamiento de las enfermedades orales se basan en el concepto erróneo de separar la salud oral de la salud general y considerar los tratamientos odontológicos como «estéticos» sin tener en cuenta que el dolor dental y la pérdida dentaria disminuyen de modo significativo la calidad de vida de las personas1. En 2003 la Organización Mundial de la Salud (OMS enfatizó la necesidad de promover la salud oral en la población mundial y publicó un documento guía para que cada país pueda definir sus metas de mejoría en los indicadores de salud oral para el año 20202.La enfermedad periodontal es una de las entidades más comunes a nivel mundial y una de las principales causas de pérdida dentaria. La gingivitis, la forma más leve de enfermedad periodontal, es ocasionada por el biofilm (placa bacteriana que se acumula en la superficie dentaria adyacente al tejido gingival (encía. La forma más severa de enfermedad periodontal es la periodontitis, secundaria como respuesta inflamatoria a la infección por gérmenes periodontales diversos (Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Treponema denticola, Tannerella forsythensis, entre otros caracterizada por la destrucción del tejido conectivo y del soporte óseo para dar lugar a la pérdida dentaria3. La Tercera Encuesta de Salud Bucal (ENSAB III que se hizo en Colombia, comunicó para todas las formas de enfermedad periodontal (gingivitis y periodontitis una prevalencia de 50.2% de