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Sample records for aeromonas caviae co-cultured

  1. Aeromonas caviae septicemia in immunocompetent gastrointestinal carriers

    Directory of Open Access Journals (Sweden)

    M. Dwivedi

    2008-12-01

    Full Text Available Aeromonas caviae strains have been isolated from blood and stool cultures of three immunocompetent patients, residents of Northern India, who presented with community acquired septicemia without any recent history of diarrhea. Cell culture infectivity test performed on Hep-2 cells have shown substantial degree of invasiveness in the isolated strains. This case unleashes a possibility of asymptomatic gastrointestinal carriage of such strains of A. caviae in a very large population of India, as several areas of India have very high rates of Aeromonas induced acute diarrhea/gastroenteritis (up to 13%. It needs to be appraised further in India as well as other countries having high rates of Aeromonas induced acute diarrhea/gastroenteritis.

  2. Aeromonas caviae septicemia in immunocompetent gastrointestinal carriers

    Directory of Open Access Journals (Sweden)

    M. Dwivedi

    Full Text Available Aeromonas caviae strains have been isolated from blood and stool cultures of three immunocompetent patients, residents of Northern India, who presented with community acquired septicemia without any recent history of diarrhea. Cell culture infectivity test performed on Hep-2 cells have shown substantial degree of invasiveness in the isolated strains. This case unleashes a possibility of asymptomatic gastrointestinal carriage of such strains of A. caviae in a very large population of India, as several areas of India have very high rates of Aeromonas induced acute diarrhea/gastroenteritis (up to 13%. It needs to be appraised further in India as well as other countries having high rates of Aeromonas induced acute diarrhea/gastroenteritis.

  3. Identification of hydroxy fatty acids in Aeromonas hydrophila, Aeromonas sobria, and Aeromonas caviae.

    OpenAIRE

    1985-01-01

    Gas-liquid chromatographic analysis of fatty acid methyl esters obtained from clinical isolates of Aeromonas hydrophila, A. sobria, and A. caviae revealed the presence of two hydroxy fatty acid species, 3-OH 12:0 and 3-OH 14:0.

  4. Aeromonas caviae strain induces Th1 cytokine response in mouse intestinal tract

    Science.gov (United States)

    Aeromonas caviae has been associated with human gastrointestinal disease. Strains of this species typically lack virulence factors (VFs) such as enterotoxins and hemolysins that are produced by other human pathogens of the Aeromonas genus,. Microarray profiling of...

  5. Draft Genome Sequence of Marine-Derived Aeromonas caviae CHZ306, a Potential Chitinase Producer Strain

    Science.gov (United States)

    Zimpel, Cristina Kraemer; Guimaraes, Ana Marcia Sa; Pessoa, Adalberto; Rivera, Irma Nelly Gutierrez

    2016-01-01

    We report here a draft genome sequence of Aeromonas caviae CHZ306, a marine-derived bacterium with the ability to hydrolyze chitin and express high levels of chitinases. The assembly resulted in 65 scaffolds with approximately 4.78 Mb. Genomic analysis revealed different genes encoding chitin-degrading enzymes that can be used for chitin derivative production. PMID:27856589

  6. DIARRHEA OUTBREAK IN PERNAMBUCO, BRAZIL, ASSOCIATED WITH A HEAT-STABLE CYTOTOXIC ENTEROTOXIN PRODUCED BY Aeromonas caviae.

    Science.gov (United States)

    Lopes, Ana Carolina Amaral; Martins, Luciano Moura; Gatti, Maria Silvia Viccari; Falavina Dos Reis, Cristhiane Moura; Hofer, Ernesto; Yano, Tomomasa

    2015-01-01

    In the present study enterotoxic and cytotoxic activities of twenty Aeromonas caviae strains were examined. They originated from fecal specimens of patients with acute diarrhea during an outbreak in Brazil in 2004. Culture supernatants of fourteen strains (70%) caused fluid accumulation in rabbit ileal intestinal loops and in suckling mice assays, and also showed a cytotoxic activity in Vero and Caco-2 cells. The enterotoxic and cytotoxic factors were heat-stable after culture supernatants treatment at 100 ºC. The results revealed that A. caviae strains produce a putative diarrheagenic virulence factor, a heat-stable cytotoxic enterotoxin that could be linked to the diarrhea outbreak that took place in Brazil.

  7. Characterisation of potential virulence markers in Aeromonas caviae isolated from polluted and unpolluted aquatic environments in Morocco

    DEFF Research Database (Denmark)

    Imziln, Boujama; Krovacek, Karel; Baloda, Suraj B.

    1998-01-01

    .406) than those isolated from raw sewage (PVI = 0.175) or those from river waters (PVI = 0.09). These results suggest that stabilisation pond systems used for sewage purification under arid climate conditions in Marrakech, may have selected potentially enteropathogenic A. caviae strains.......A total of 100 Aeromonas caviae strains isolated from river waters (38 isolates), raw sewage (30 isolates) and effluents of stabilisation ponds (i.e. treated sewage; 32 isolates) in Marrakech, Morocco, were tested for the presence of putative virulence factors to delineate differences, if any......, in their enteropathogenic potential in relation to the source of isolation. A number of A. caviae isolates were able to elaborate at least one of the tested virulence factors. Of the 100 strains tested, 27 were capable of producing haemolysins, 19 produced cytotoxin, 24 produced cytotonic toxin, while 18 of the isolates...

  8. Identification of clinical aeromonas species by rpoB and gyrB sequencing and development of a multiplex PCR method for detection of Aeromonas hydrophila, A. caviae, A. veronii, and A. media.

    Science.gov (United States)

    Persson, Søren; Al-Shuweli, Suzan; Yapici, Seval; Jensen, Joan N; Olsen, Katharina E P

    2015-02-01

    Conventional identification of Aeromonas species based on biochemical methods is challenged by the heterogeneous nature of the species. Here, we present a new multiplex PCR method directed toward the gyrB and rpoB genes that identifies four Aeromonas species, A. hydrophila, A. media, A. veronii, and A. caviae, and we describe the application of this method on a Danish strain collection.

  9. Cloning, sequencing and expression of the gene encoding the extracellular metalloprotease of Aeromonas caviae.

    Science.gov (United States)

    Kawakami, K; Toma, C; Honma, Y

    2000-01-01

    A gene (apk) encoding the extracellular protease of Aeromonas caviae Ae6 has been cloned and sequenced. For cloning the gene, the DNA genomic library was screened using skim milk LB agar. One clone harboring plasmid pKK3 was selected for sequencing. Nucleotide sequencing of the 3.5 kb region of pKK3 revealed a single open reading frame (ORF) of 1,785 bp encoding 595 amino acids. The deduced polypeptide contained a putative 16-amino acid signal peptide followed by a large propeptide. The N-terminal amino acid sequence of purified recombinant protein (APK) was consistent with the DNA sequence. This result suggested a mature protein of 412 amino acids with a molecular mass of 44 kDa. However, the molecular mass of purified recombinant APK revealed 34 kDa by SDS-PAGE, suggesting that further processing at the C-terminal region took place. The 2 motifs of zinc binding sites deduced are highly conserved in the APK as well as in other zinc metalloproteases including Vibrio proteolyticus neutral protease, Emp V from Vibrio vulnificus, HA/P from Vibrio cholerae, and Pseudomonas aeruginosa elastase. Proteolytic activity was inhibited by EDTA, Zincov, 1,10-phenanthroline and tetraethylenepentamine while unaffected by the other inhibitors tested. The protease showed maximum activity at pH 7.0 and was inactivated by heating at 80 C for 15 min. These results together suggest that APK belongs to the thermolysin family of metalloendopeptidases.

  10. An Aeromonas caviae Genomic Island Is Required for both O-Antigen Lipopolysaccharide Biosynthesis and Flagellin Glycosylation ▿

    OpenAIRE

    Tabei, S. Mohammed B.; Hitchen, Paul G.; Day-Williams, Michaela J.; Merino, Susana; Vart, Richard; Pang, Poh-Choo; Horsburgh, Gavin J.; Viches, Silvia; Wilhelms, Markus; Tomás, Juan M.; Dell, Anne; Shaw, Jonathan G

    2009-01-01

    Aeromonas caviae Sch3N possesses a small genomic island that is involved in both flagellin glycosylation and lipopolysaccharide (LPS) O-antigen biosynthesis. This island appears to have been laterally acquired as it is flanked by insertion element-like sequences and has a much lower G+C content than the average aeromonad G+C content. Most of the gene products encoded by the island are orthologues of proteins that have been shown to be involved in pseudaminic acid biosynthesis and flagellin gl...

  11. PARTICIPACION DEL COMPLEJO PIRUVATO DESHIDROGENASA DE AEROMONAS CAVIAE ST EN RESISTENCIA TELURITO DE POSTASIO (K2TEO3)

    OpenAIRE

    2008-01-01

    El telurito de potasio (K2TeO3) es altamente tóxico para la mayoría de los seres vivos. Aunque se desconoce el mecanismo de toxicidad especifico, recientemente se ha demostrado que estaría dado, al menos en parte, por el establecimiento de un estrés oxidativo. Aeromonas caviae ST, el microorganismo utilizado en este estudio es altamente resistente a K2TeO3 y extractos crudos de esta bacteria catalizan la reducción de K2TeO3 in vitro. Se obtuvo fracciones con actividad telurito reductasa qu...

  12. Aeromonas caviae alters the cytosolic and mitochondrial creatine kinase activities in experimentally infected silver catfish: Impairment on renal bioenergetics.

    Science.gov (United States)

    Baldissera, Matheus D; Souza, Carine F; Júnior, Guerino B; Verdi, Camila Marina; Moreira, Karen L S; da Rocha, Maria Izabel U M; da Veiga, Marcelo L; Santos, Roberto C V; Vizzotto, Bruno S; Baldisserotto, Bernardo

    2017-09-01

    Cytosolic and mitochondrial creatine kinases (CK), through the creatine kinase-phosphocreatine (CK/PCr) system, provide a temporal and spatial energy buffer to maintain cellular energy homeostasis. However, the effects of bacterial infections on the kidney remain poorly understood and are limited only to histopathological analyses. Thus, the aim of this study was to investigate the involvement of cytosolic and mitochondrial CK activities in renal energetic homeostasis in silver catfish experimentally infected with Aeromonas caviae. Cytosolic CK activity decreased in infected animals, while mitochondrial CK activity increased compared to uninfected animals. Moreover, the activity of the sodium-potassium pump (Na(+), K(+)-ATPase) decreased in infected animals compared to uninfected animals. Based on this evidence, it can be concluded that the inhibition of cytosolic CK activity by A. caviae causes an impairment on renal energy homeostasis through the depletion of adenosine triphosphate (ATP) levels. This contributes to the inhibition of Na(+), K(+)-ATPase activity, although the mitochondrial CK activity acted in an attempt to restore the cytosolic ATP levels through a feedback mechanism. In summary, A. caviae infection causes a severe energetic imbalance in infected silver catfish, which may contribute to disease pathogenesis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Draft Genome Sequence of Aeromonas caviae CH129, a Marine-Derived Bacterium Isolated from the Coast of São Paulo State, Brazil

    Science.gov (United States)

    Alfonso Vargas, Nadia Catalina; Zimpel, Cristina Kraemer; Pessoa, Adalberto; Rivera, Irma Nelly Gutierrez

    2016-01-01

    We report here the draft genome sequence of Aeromonas caviae CH129, a marine-derived bacterium isolated from the coast of São Paulo state, Brazil. Genomic analysis revealed genes encoding enzymes involved in binding, transport, and chitin metabolism and different virulence-associated factors. PMID:27908996

  14. Emergence of CTX-M-3, TEM-1 and a new plasmid-mediated MOX-4 AmpC in a multiresistant Aeromonas caviae isolate from a patient with pneumonia.

    Science.gov (United States)

    Ye, Ying; Xu, Xi-Hai; Li, Jia-Bin

    2010-07-01

    Aeromonas species rarely cause pulmonary infection. We report, for what is believed to be the first time, a case of severe pneumonia in a cancer patient caused by Aeromonas caviae. Detailed microbiological investigation revealed that this isolate carried three beta-lactamase-encoding genes (encoding MOX-4, CTX-M-3 and TEM-1) conferring resistance to all beta-lactams but imipenem. The beta-lactamase with a pI of 9.0 was transferred by conjugation and associated with a 7.3 kb plasmid, as demonstrated by Southern blot hybridization. Analysis of the nucleotide and amino acid sequences showed a new ampC gene that was closely related to those encoding the MOX-1, MOX-2 and MOX-3 beta-lactamases. This new plasmid-mediated AmpC beta-lactamase from China was named MOX-4. This is believed to be the first report of MOX-4, CTX-M-3 and TEM-1 beta-lactamases in a multiresistant A. caviae.

  15. Distribution of Aeromonas Species in the Intestinal Tracts of River Fish

    OpenAIRE

    1995-01-01

    Aeromonas isolates were obtained from fish intestines, water, and sediments from an urban river and identified by the DNA-DNA microplate hybridization method. The isolates were Aeromonas veronii (22%), Aeromonas caviae (18%), Aeromonas hydrophila (13%), Aeromonas sobria (8%), Aeromonas jandaei (7%), and other Aeromonas spp. (33%). Aeromonas species occurred at high densities with high incidences, regardless of season. The results strongly suggest that aeromonads are indigenous in fish intesti...

  16. Indirect Enzyme-Linked Immunosorbent Assay (ELISA) for Rapid Detection of Aeromonas caviae%豚鼠气单胞菌快速检测间接ELISA法的建立

    Institute of Scientific and Technical Information of China (English)

    吴斌

    2006-01-01

    应用从患败血症的鳗鲡中分离、纯化出的豚鼠气单胞菌(Aeromonas caviae)菌株AB40511NA1,制备了兔抗血清,建立了快速检测豚鼠气单胞菌的间接ELISA方法.研究结果显示:AB40511NA1兔抗血清的效价为1:16000.该法对AB40511NA1最低检测量为6×104 cfu·mL-1;与水产养殖动物主要致病菌嗜水气单胞菌(A. hydrophila)、温和气单胞菌(A. sobria)、鳗弧菌(V. anguillarum)、溶藻弧菌(V.alginolyticus)、副溶血弧菌(V. parahaemolyticus)及非O1-群霍乱弧菌(Non-O1 Vibrio cholerae)均无交叉反应,特异性强;能快速、准确、方便地检测出豚鼠气单胞菌.

  17. Comparative in vitro activities of selected antimicrobial agents against Aeromonas species and Plesiomonas shigelloides.

    OpenAIRE

    1985-01-01

    The in vitro antimicrobial susceptibilities of fecal isolates of Aeromonas caviae, Aeromonas hydrophila, Aeromonas sobria, and Plesiomonas shigelloides were tested by agar dilution. This in vitro study suggested that ciprofloxacin, enoxacin, norfloxacin, tetracycline, and trimethoprim-sulfamethoxazole might be effective oral agents for treatment of diarrhea involving Aeromonas spp. or P. shigelloides.

  18. MONITORING THE EFFECTIVENESS OF UV DISINFECTION OF AEROMONAS SPP. USING SELECTIVE AND NON-SELECTIVE MEDIA

    Science.gov (United States)

    This research was initiated to determine the sensitivity of Aeromonas spp. to ultraviolet (UV) disinfection. Aeromonas hydrophila is a contaminant listed on the USEPA's 1998 CCL. Three different Aeromonas spp. (A. hydrophila, A. sobria and A. caviae) were tested using membrane fi...

  19. CHARACTERIZATION OF AEROMONAS VIRULENCE USING AN IMMUNOCOMPROMISED MOUSE MODEL

    Science.gov (United States)

    An immunocompromised mouse model was used to characterize Aeromonas strains for their ability to cause opportunistic, extraintestinal infections. A total of 34 isolates of Aeromonas (A. hydrophila [n = 12]), A. veronii biotype sobria [n = 7], A. caviae [n = 4], A. enchelia [n = 4...

  20. Inhibitory effects of sixteen kinds of Chinese herbal medicine and their compounds on pathogenic Aeromonas caviae from cultured Andrias davidianus in vitro%十六种中草药及其复方对大鲵致病性豚鼠气单胞菌的体外抑制作用

    Institute of Scientific and Technical Information of China (English)

    凌空; 王涵; 丁诗华; 吴兴镇

    2015-01-01

    To study the inhibitory effects of sixteen kinds of Chinese herbal medicine and their compounds on pathogenic Aeromonas caviae from cultured Andrias davidianus in vitro, the minimum inhibitory concentration (MIC) and the mini-mum bactericidal concentration (MBC) of single drug and the compounds on pathogenic A.caviae from A.davidianus were measured with agar double dilution method.Then sixteen kinds of chinese herbal medicine were mixed one by one based on chessboard experimental method to get one hundred and twenty kinds of compounds of Chinese herbal medicine .The results showed that sixteen kinds of Chinese herbal medicine had antibacterial effect at certain extent on the pathogenic bacteri -a.Terminalia chebula Retz and Rhus chinensis Mil had the best antibacterial activity, followed by Pomegranate Rind, Rheum palmatum L, Sophora japonica L, Rubus idaeus, Scutellaria baicalensis, Rosa laevigata, Cistanche deserticola and Syringa Linn, Radix Cyathulae, Paeonia veitchii, Anemarrhena asphodeloides, DivaricateSaposhnikovia Root, Musk-root Semiaquilegia Root and Momordica grosvenori had the worst antibacterial activity.The study indicated that the rational use of Chinese herbal medicine could increase the inhibitory effects of the drug on A.caviae from A.davidianus in vitro drug Antibacterial activity.%应用二倍稀释法测定了诃子、五倍子、石榴皮等16种中草药及其复方对大鲵(Andrias davidianus)致病性豚鼠气单胞菌(Aeromonas caviae)的最小抑菌浓度和最小杀菌浓度,应用棋盘法设计16种中草药组合的120种双联复方和10种中草药组合的120种三联复方,评估了复方的体外抑菌作用。结果表明:16种中草药对大鲵致病性豚鼠气单胞菌均有一定程度的抑菌作用,抑菌作用强弱依次为诃子、五倍子、石榴皮、大黄、槐角、覆盆子、黄芩、金樱子、肉苁蓉、丁香、川牛膝、赤芍、知母、防风、天葵子及罗汉果;在120个中

  1. Hemolytic Activity and Siderophore Production in Different Aeromonas Species Isolated from Fish

    OpenAIRE

    Santos, Jesús A.; González, César J.; Otero, Andrés; García-López, María-Luisa

    1999-01-01

    The hemolytic activity and siderophore production of several strains of motile aeromonads were determined. The hemolytic activity of Aeromonas caviae and Aeromonas eucrenophila was enhanced after trypsinization of the samples. The enhancement of hemolysis was observed in strains that carried an aerolysin-like gene, detected by a PCR procedure. Siderophore production was demonstrated in all but one strain of Aeromonas jandaei. No apparent relationship was observed between the presence of plasm...

  2. A prospective study on Aeromonas in outpatients with diarrhea in the central region of Rio Grande do Sul State

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    Karoline de Campos Prediger

    2012-09-01

    Full Text Available Aeromonas spp. were identified in five (2,7% of 182 diarrheal stool cultures, A. caviae was predominant, resistant mainly to ampicillin and cephalotin. This is the first study showing the presence of Aeromonas spp. in diarrheal stools of outpatients in the central region of Rio Grande do Sul State, Brazil.

  3. Clinical implications of species identification in monomicrobial Aeromonas bacteremia.

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    Chi-Jung Wu

    Full Text Available Advances in Aeromonas taxonomy have led to the reclassification of aeromonads. Hereon, we aimed to re-evaluate the characteristics of Aeromonas bacteremia, including those of a novel species, Aeromonas dhakensis.A retrospective study of monomicrobial Aeromonas bacteremia at a medical center in southern Taiwan from 2004-2011 was conducted. Species identification was based on rpoB sequencing. Of bacteremia of 153 eligible patients, A. veronii (50 isolates, 32.7%, A. dhakensis (48, 31.4%, A. caviae (43, 28.1%, and A. hydrophila (10, 6.5% were the principal causative species. A. dhakensis and A. veronii bacteremia were mainly community-acquired and presented as primary bacteremia, spontaneous bacterial peritonitis, or skin and soft-tissue infection, whereas A. caviae was associated with hospital-onset bacteremia. The distribution of the AmpC β-lactamase and metallo-β-lactamase genes was species-specific: bla(AQU-1, bla(MOX, or bla(CepH was present in A. dhakensis, A. caviae, or A. hydrophila, respectively, and bla(CphA was present in A. veronii, A. dhakensis, and A. hydrophila. The cefotaxime resistance rates of the A. caviae, A. dhakensis, and A. hydrophila isolates were higher than that of A. veronii (39.5%%, 25.0%, and 30% vs. 2%, respectively. A. dhakensis bacteremia was linked to the highest 14-day sepsis-related mortality rate, followed by A. hydrophila, A. veronii, and A. caviae bacteremia (25.5%, 22.2%, 14.0%, and 4.7%, respectively; P = 0.048. Multivariate analysis revealed that A. dhakensis bacteremia, active malignancies, and a Pitt bacteremia score ≥ 4 was an independent mortality risk factor.Characteristics of Aeromonas bacteremia vary between species. A. dhakensis prevalence and its associated poor outcomes suggest it an important human pathogen.

  4. Correlation of the Suicide Phenomenon in Aeromonas Species with Virulence and Enteropathogenicity

    OpenAIRE

    1989-01-01

    Certain strains of mesophilic aeromonads (Aeromonas hydrophila, A. sorbria, and A. caviae), when grown in broth containing 0.5% glucose, undergo growth inhibition concomitant with acetate accumulation. Because these strains are nonviable after 24 h, this phenomenon is termed suicide. We investigated suicidal strains of Aeromonas species as a means of understanding animal virulence and enteropathogenicity. To assess virulence, batches of five white mice were inoculated intraperitoneally with 1...

  5. Genotypic and phenotypic identification of Aeromonas species and CphA-mediated carbapenem resistance in Queensland, Australia.

    Science.gov (United States)

    Sinclair, Holly A; Heney, Claire; Sidjabat, Hanna E; George, Narelle M; Bergh, Haakon; Anuj, Snehal N; Nimmo, Graeme R; Paterson, David L

    2016-05-01

    Infection caused by Aeromonas spp. ranges from superficial wound infection to life-threatening septicemia. Carbapenem resistance due to metallo-beta-lactamase, CphA encoded by the cphA gene, is a significant problem. This study defines Aeromonas spp. causing clinical disease in Queensland, Australia. Phenotypic tests for carbapenemase detection were assessed. One hundred Aeromonas isolates from blood (22), wound (46), sterile sites (11), stool (18), eye (2), and sputum (1) were characterized by rpoB and gyrB sequencing. Meropenem susceptibility by VITEK2, disk diffusion, and E-test MIC were determined. Carbapenemase production was assessed by Carba NP test and cphA by PCR. Gene sequencing identified isolates as Aeromonas dhakensis (39), Aeromonas veronii (21), Aeromonas hydrophila (20), Aeromonas caviae (14), Aeromonas jandaei (4), Aeromonas bestiarum (1), and Aeromonas sanarellii (1). Disk diffusion and E-test failed to detect resistance in isolates with presence of cphA. Carba NP was performed with 97.4% sensitivity and 95.7% specificity. Carbapenem resistance gene cphA was detected in A. veronii (21; 100%), A. hydrophila (18; 90%), A. dhakensis (34; 87.2%), A. jandaei (3; 75%), and A. bestiarum (1; 100%) but not A. caviae. We found that A. dhakensis was the predominant species, a previously unrecognized pathogen in this region.

  6. Isolation and antibiotic sensitivity of Aeromonas from children with diarrhea

    Directory of Open Access Journals (Sweden)

    Meiyanti Meiyanti

    2016-02-01

    Full Text Available Aeromonas species are gram-negative, motile, facultative anaerobic, rod shaped, oxidase positive bacteria of the recently assigned family Aeromonadaceae. The significance of Aeromonas species as causative agent of human diarrhoea has recently been established. The aim of the present study was to investigate the distribution, and antibiotic sensitivity of Aeromonas in nonhospitalized children with diarrhea.One hundred and seventeen rectal swabs from children with diarhhea were cultured for isolation of Aeromonas organisms as the etiological agents. In addition to Aeromonas, other enteric pathogens were also isolated. Overall, the isolates of enteric pathogens amounted to 36.8%, consisting of Salmonella, Shigella, Aeromonas, and Vibrio. Aeromonas was only found in 5.1% of cultures, with a ratio of A. caviae and A. hydrophila of 2:1, while Salmonella made up the majority of causative organisms with an isolation frequency of 18.8%, followed by Shigella with 11.1%. In this study no isolates of Vibrio cholerae O1 were found as etiological agents of diarrhea; however, V. cholerae non-O1 and V. parahaemolyticus were found in small numbers (<1%. All isolates of Aeromonas were resistant to ampicillin and tetracycline, but sensitive to ciprofloxacin and ceftriaxone, as were the other enteric pathogens. Although the frequency of isolation of these enteric pathogens was higher than for Vibrio spp., their role in infective diarrhea was less clearcut in comparison with Salmonella and Shigella.

  7. Biliary tract infections caused by Aeromonas species.

    Science.gov (United States)

    Chao, C M; Lai, C C; Tang, H J; Ko, W C; Hsueh, P-R

    2013-02-01

    This study investigated the clinical and microbiological characteristics of patients with Aeromonas infections of the biliary tract. Patients with bile cultures positive for Aeromonas species during the period July 2004 to December 2011 were identified from a computerized database of a hospital in Taiwan. Patients with Aeromonas infections of the biliary tract were further identified. During the study period, a total of 1,142 isolates of Aeromonas species were obtained from 750 patients. Of those patients, 91 (12.1 %) had Aeromonas infections of the biliary tract. The annual incidence (episodes per 10,000 patient-days) of biliary tract infections caused by all Aeromonas species was 0.31 in 2007, 0.12 in 2010, and 0.27 in 2011. A. hydrophila was the most common species isolated (n = 41, 45.1 %), followed by A. caviae (n = 30, 33.0 %), A. veronii biovar sobria (n = 15, 16.5 %), and A. veronii biovar veronii (n = 5, 5.5 %). The majority of patients (n = 77, 84.6 %) had polymicrobial infections. Hepatobiliary stones (n = 50, 54.9 %) and hepatobiliary cancer (n = 38, 41.8 %) were the most common underlying diseases, followed by diabetes mellitus (n = 29, 31.9 %) and liver cirrhosis (n = 7, 7.7 %). The in-hospital mortality rate was 8.8 %. Infection-related mortality was associated with underlying immunocompromised condition (p = 0.044) and use of mechanical ventilation (p = 0.004), but was not associated with inappropriate antibiotic usage or concomitant bacteremia (n = 8, 8.8 %). In conclusion, biliary tract infections caused by Aeromonas species are not uncommon and can develop in both immunocompromised and immunocompetent patients; however, patients with underlying hepatobiliary diseases are particularly susceptible to these infections.

  8. Incidence and virulence characteristics of Aeromonas spp. in fish

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    Ashraf M. Abd-El-Malek

    2017-01-01

    Full Text Available Aim: This study was conducted to evaluate the presence of Aeromonas spp. in raw and ready-to-eat (RTE fish commonly consumed in Assiut city, Egypt, and to determine virulence factors due to they play a key role in their pathogenicity. Materials and Methods: A total of 125 samples of raw and RTE fish samples were taken from different fish markets and fish restaurants in Assiut Governorate and screened for the presence of Aeromonas spp. by enrichment on tryptic soy broth then incubated at 30°C for 24 h. Plating unto the sterile Petri dishes containing Aeromonas agar base to which Aeromonas selective supplement was added. The plates were incubated at 37°C for 24 h. Presumptive Aeromonas colonies were biochemically confirmed and analyzed for pathogenicity by hemolysin production, protease, and lipase detection. Results: The results indicated that raw fish were contaminated with Aeromonas spp. (40% in wild and 36% in cultured Nile tilapia. Regarding RTE, Aeromonas spp. could be isolated with the percentage of 16%, 28% and 20% in fried Bolti, grilled Bolti and fried Bayad, respectively. Out of 35 isolates obtained, 22 were categorized as Aeromonas hydrophila, 12 were classified as Aeromonas sobria and Aeromonas caviae were found in only one isolate. The virulence factors of Aeromonas spp. were detected and the results showed that all isolates produced of hemolysin (91.4%, protease (77.1%, and lipase enzyme (17.1%. Conclusion: This study indicates that the presence of A. hydrophila with virulence potential in fresh and RTE fish may be a major threat to public health.

  9. 南方鲇(Stlurusmeridionalis)豚鼠气单胞茵溶血素的分离、纯化与致病性研究%ISOLATION, PURIFICATION AND PATHOGENICITY OF THE HAEMOLYSIN PRODUCED BY AEROMONAS CAVIAE FROM SILURUS MERIDIONALIS CHEN

    Institute of Scientific and Technical Information of China (English)

    汪开毓; 肖丹; 贺扬; 陈德芳; 耿毅; 刘天强; 黄冠军

    2012-01-01

    采用硫酸铵盐析、DEAE.SephadexA50凝胶层析法对豚鼠气单胞菌的溶血素进行分离、纯化。获得分子量为32.2kDa的单一多肽溶血素,测定溶血价为2^5。将该溶血素倍比稀释后梯度接种南方鲇,研究其对南;h-鲇的致病性和组织病理损伤情况。结果表明:豚鼠气单胞菌溶血素具有较强的毒力,对南方鲇的半数致死剂量(LD,0)为0.29mg蛋白/k体重;病鱼表现为体表褪色,腹部膨大,肝、脾、肾肿大,肝、脾淤血、出血明显;胃肠道内充有大量粘液,黏膜充血、出血;组织病理学表现为骨骼肌水肿、变性,肝细胞变性、坏死,血窦扩张淤血,肾小,管上皮细胞变性、坏死,脾脏淋巴细胞减少,胃肠道黏膜上皮变性、坏死,脱落。%The haemolysin produced by Aeromonas caviae was salted out by ammonium sulphate precipitation and purified by chromatography on Sephadex A50. It was estimated to a homogeneous polypeptide of 32.2kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Haemolysis assay showed that its haemolytic activity was 25. To investigate its pathogenicity, double dilution haemolysin was intraperitoneally injected into Silurus meridionalis Chen. The resuits showed that the purified haemolysin was toxic towards S. meridionalis, with a half-lethal dose (LDso) of 0.29mg/kg (protein/weight). Fish injected with this haemolysin exhibited symptoms of color fading, abdominal intumescence, congestion and enlargement of liver and spleen, renomegaly, gastrointestinal inflammation. Skeletal muscle appeared swollen and disruption with red stained sarcoplasm. Liver corroded to cell death and necrosis, and were congested with few sinusoid. Degeneration and necrosis was found in the renal cells, which could extend to some histolysis. Renal interstitium showed a reduced incidence of lymphocytes with numerous inflammatory cells infiltration. Spleen lymphocytes were reduction

  10. Molecular characterization of fluoroquinolone-resistant Aeromonas spp. isolated from imported shrimp.

    Science.gov (United States)

    Shakir, Zakiya; Khan, Saeed; Sung, Kidon; Khare, Sangeeta; Khan, Ashraf; Steele, Roger; Nawaz, Mohamed

    2012-11-01

    Sixty-three nalidixic acid-resistant Aeromonas sp. isolates were obtained from imported shrimp. Phylogenetic analysis of gyrB sequences indicated that 18 were A. enteropelogenes, 26 were A. caviae, and 19 were A. sobria. Double missense mutations in the quinolone resistance-determining region (QRDR) of gyrA at codon 83 (Ser→Val/Ile) and codon 92 (Leu→Met) coupled with a point mutation of parC at codon 80 (Ser→Ile/Phe) conferred high levels of quinolone resistance in the isolates. A majority of A. enteropelogenes and A. caviae strains harbored toxin genes, whereas only a few A. sobria strains harbored these genes. The fluoroquinolone-resistant Aeromonas spp. exhibited higher cytotoxicity than fluoroquinolone-sensitive, virulent Aeromonas spp. to rat epithelial cells.

  11. Reassessment of the Enteropathogenicity of Mesophilic Aeromonas Species

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    Teunis, Peter; Figueras, Maria J.

    2016-01-01

    Cases of Aeromonas diarrhea have been described all over the world. The genus Aeromonas includes ca. 30 species, of which 10 have been isolated in association with gastroenteritis. The dominating species that account for ca. 96% of the identified strains are Aeromonas caviae, A. veronii, A. dhakensis, and A. hydrophila. However, the role of Aeromonas as a true enteropathogen has been questioned on the basis of the lack of outbreaks, the non-fulfillment of Koch’s postulates and the low numbers of acute illnesses in the only existing human challenge study. In the present study we reassess the enteropathogenicity of Aeromonas using dose response models for microbial infection and acute illness. The analysis uses the data from the human challenge study and additional data from selected outbreak investigations where the numbers exposed and the dose were reported, allowing their inclusion as “natural experiments”. In the challenge study several cases of asymptomatic shedding were found (26.3%, 15/57), however, only 3.5% (2/57) of those challenged with Aeromonas developed acute enteric symptoms (i.e., diarrhea). The “natural experiments” showed a much higher risk of illness associated with exposure to Aeromonas, even at moderate to low doses. The median dose required for 1% illness risk, was ~1.4 × 104 times higher in the challenge study (1.24 × 104 cfu) compared to natural exposure events (0.9 cfu). The dose response assessment presented in this study shows that the combined challenge and outbreak data are consistent with high infectivity of Aeromonas, and a wide range of susceptibility to acute enteric illness. To illustrate the outcomes, we simulate the risk associated with concentrations of Aeromonas found in different water and food matrices, indicating the disease burden potentially associated with these bacteria. In conclusion this study showed that Aeromonas is highly infectious, and that human susceptibility to illness may be high, similar to

  12. AISLADA DE Cavia porcellus

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    H. Jurado-Gámez

    2014-01-01

    Full Text Available Se evaluó el efectoin vitrodeLactobacillus plantarumsobreYersinia pseudotuberculosisaislada de cuyes (Cavia porcellus e identificada bioquímica y molecularmente. Se deter-minó la viabilidad deL. plantaruma diferentes pH (3.25, 4.5, 7.6 y 8.2, sales biliaresbovinas (3, 4 y 5%, bilis bovina (0.5, 1 y 2% y temperatura (38 y 45°C. Se realizóantibiograma a ambas bacterias con Cefalotina, Cefepima, Ciprofloxacina, Dicloxacilina,Enrofloxacina, Gentamicina, Penicilina y Trimetropim-Sulfametoxasol. Se determina-ron los péptidos deL. plantarummediante HPLC y la cinética de crecimiento en dossustratos (MRS y Pro, midiéndose: biomasa, pH, consumo de azúcar total y ácidoláctico. Se observó un mejor crecimiento bajo pH 3.25, sales biliares 5%, bilis bovina2% y temperatura de 45°C con valores de 3.0·1012, 2.3·1012, 8.6·1011y 3.0·1012UFC/ml, respectivamente.L. plantarumy el sobrenadante inhibieron aY. pseudotuberculosis,con halos de 3 y 5 mm. Se observó resistencia deL. plantarumyY. pseudotuberculosisaPenicilina y Dicloxacilina, respectivamente, con sensibilidad de ambas cepas a los demásantibióticos. Se evidenciaron los péptidos VAL-TIR-VAL y TIR-GLI-GLI-FA-METen el sobrenadante deL. plantarum. Durante la fase logarítmica para los medios MRSy Pro se alcanzaron valores de 6.75·1012y 1.20·1012UFC/ml (biomasa, 6.96 y 5.49(pH, y 0.65 y 0.76%, (ácido láctico, respectivamente. La comparación de los mediosmediante un diseño de bloques al azar (P> 0.05, mostró similar comportamiento.L.plantarumdemostró su potencial probiótico en condicionesin vitro.

  13. Effectiveness of radiation processing in elimination of Aeromonas from food

    Energy Technology Data Exchange (ETDEWEB)

    Nagar, Vandan [Food Technology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Bandekar, Jayant R., E-mail: jrb@barc.gov.i [Food Technology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)

    2011-08-15

    Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Contaminated water, sprouts, vegetables, seafood and food of animal origin have been considered to be the important sources of Aeromonas infection. In the present study, radiation sensitivity of indigenous strains of Aeromonas spp. from different food samples was evaluated. The decimal reduction dose (D{sub 10}) values of different Aeromonas isolates in saline at 0-4 {sup o}C were in the range of 0.031-0.046 kGy. The mixed sprouts, chicken and fish samples were inoculated with a cocktail of five most resistant isolates (A. salmonicida Y567, A. caviae A85, A. jandaei A514A, A. hydrophila CECT 839{sup T} and A. veronii Y47) and exposed to {gamma} radiation to study the effectiveness of radiation treatment in elimination of Aeromonas. D{sub 10} values of Aeromonas cocktail in mixed sprouts, chicken and fish samples were found to be 0.081{+-}0.001, 0.089{+-}0.003 and 0.091{+-}0.003 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in complete elimination of 10{sup 5} CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples. No recovery of Aeromonas was observed in the 1.5 kGy treated samples stored at 4 {sup o}C up to 12 (mixed sprouts) and 7 days (chicken and fish samples), even after enrichment and selective plating. This study demonstrates that a 1.5 kGy dose of irradiation treatment could result in complete elimination of 10{sup 5} CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples.

  14. Effectiveness of radiation processing in elimination of Aeromonas from food

    Science.gov (United States)

    Nagar, Vandan; Bandekar, Jayant R.

    2011-08-01

    Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Contaminated water, sprouts, vegetables, seafood and food of animal origin have been considered to be the important sources of Aeromonas infection. In the present study, radiation sensitivity of indigenous strains of Aeromonas spp. from different food samples was evaluated. The decimal reduction dose (D10) values of different Aeromonas isolates in saline at 0-4 °C were in the range of 0.031-0.046 kGy. The mixed sprouts, chicken and fish samples were inoculated with a cocktail of five most resistant isolates (A. salmonicida Y567, A. caviae A85, A. jandaei A514A, A. hydrophila CECT 839T and A. veronii Y47) and exposed to γ radiation to study the effectiveness of radiation treatment in elimination of Aeromonas. D10 values of Aeromonas cocktail in mixed sprouts, chicken and fish samples were found to be 0.081±0.001, 0.089±0.003 and 0.091±0.003 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in complete elimination of 105 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples. No recovery of Aeromonas was observed in the 1.5 kGy treated samples stored at 4 °C up to 12 (mixed sprouts) and 7 days (chicken and fish samples), even after enrichment and selective plating. This study demonstrates that a 1.5 kGy dose of irradiation treatment could result in complete elimination of 105 CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples.

  15. Skin and soft-tissue infections caused by Aeromonas species.

    Science.gov (United States)

    Chao, C M; Lai, C C; Tang, H J; Ko, W C; Hsueh, P-R

    2013-04-01

    This study investigated the clinical characteristics of patients with skin and soft-tissue infections (SSTIs) due to Aeromonas species. Patients with SSTIs caused by Aeromonas species during the period from January 2009 to December 2011 were identified from a computerized database of a regional hospital in southern Taiwan. The medical records of these patients were retrospectively reviewed. A total of 129 patients with SSTIs due to Aeromonas species were identified. A. hydrophila (n = 77, 59.7 %) was the most common pathogen, followed by A. veronii biovar sobria (n = 22, 17.1 %), A. veronii biovar veronii (n = 20, 15.5 %), A. caviae (n = 9, 7.0 %), and A. schubertii (n = 1, 0.8 %). The most common isolates obtained from patients with polymicrobial infections were Klebsiella species (n = 33), followed by Enterococcus spp. (n = 24), Enterobacter spp. (n = 21), Escherichia coli (n = 17), Staphylococcus spp. (n = 17), Streptococcus spp. (n = 17), and Acinetobacter spp. (n = 15). Liver cirrhosis and concomitant bacteremia were more common among patients with monomicrobial Aeromonas SSTIs than among patients with polymicrobial SSTIs. Nine (7 %) patients required limb amputations. The in-hospital mortality rate was 1.6 %. In conclusion, Aeromonas species should be considered as important causative pathogens of SSTIs, and most infections are polymicrobial. In addition, the clinical presentation differs markedly between patients with monomicrobial and those with polymicrobial Aeromonas SSTIs.

  16. Virulence diversity among bacteremic Aeromonas isolates: ex vivo, animal, and clinical evidences.

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    Po-Lin Chen

    Full Text Available BACKGROUND: The objective of this study was to compare virulence among different Aeromonas species causing bloodstream infections. METHODOLOGY/PRINCIPAL FINDINGS: Nine of four species of Aeromonas blood isolates, including A. dhakensis, A. hydrophila, A. veronii and A. caviae were randomly selected for analysis. The species was identified by the DNA sequence matching of rpoD. Clinically, the patients with A. dhakensis bacteremia had a higher sepsis-related mortality rate than those with other species (37.5% vs. 0%, P = 0.028. Virulence of different Aeromonas species were tested in C. elegans, mouse fibroblast C2C12 cell line and BALB/c mice models. C. elegans fed with A. dhakensis and A. caviae had the lowest and highest survival rates compared with other species, respectively (all P values <0.0001. A. dhakensis isolates also exhibited more cytotoxicity in C2C12 cell line (all P values <0.0001. Fourteen-day survival rate of mice intramuscularly inoculated with A. dhakensis was lower than that of other species (all P values <0.0001. Hemolytic activity and several virulence factor genes were rarely detected in the A. caviae isolates. CONCLUSIONS/SIGNIFICANCE: Clinical data, ex vivo experiments, and animal studies suggest there is virulence variation among clinically important Aeromonas species.

  17. Chironomid egg masses harbour the clinical species Aeromonas taiwanensis and Aeromonas sanarellii.

    Science.gov (United States)

    Beaz-Hidalgo, Roxana; Shakèd, Tamar; Laviad, Sivan; Halpern, Malka; Figueras, María J

    2012-12-01

    Bacteria of the genus Aeromonas are found worldwide in aquatic environments and may produce human infections. In 2010, two new clinical species, Aeromonas sanarellii and Aeromonas taiwanensis, were described on the basis of one strain recovered from wounds of hospitalized patients in Taiwan. So far, only four environmental isolates of A. sanarellii and one of A. taiwanensis have been recorded from waste water in Portugal and an additional clinical strain of A. taiwanensis from the faeces of a patient with diarrhoea in Israel. In the present study, strains belonging to these two species were identified from chironomid egg masses from the same area in Israel by sequencing the rpoD gene. This represents a new environmental habitat for these novel species. The first data on the virulence genes and antibiotic susceptibility are provided. The isolates of these two new species possess multiple virulence genes and are sensitive to amikacin, aztreonam, cefepime, cefoxatime, ceftazidime, ciprofloxacin, gentamicin, piperacillin-tazobactam, tigecycline, tobramycin, trimethoprim-sulfamethoxazole and imipenem. The key phenotypic tests for the differentiation of these new species from their closest relative Aeromonas caviae included the utilization of citrate, growth at 45 °C in sheep blood agar and acid production of cellobiose.

  18. Aeromonas in Arab countries: 1995-2014.

    Science.gov (United States)

    Ghenghesh, Khalifa Sifaw; Rahouma, Amal; Zorgani, Abdulaziz; Tawil, Khaled; Al Tomi, Abdurazzaq; Franka, Ezzadin

    2015-10-01

    The aim of this review is to provide information on the prevalence, clinical syndromes, and antimicrobial resistance and therapy of Aeromonas spp. infections in Arab countries. The data were obtained by an English language literature search from 1995 to 2014 of Medline and PubMed for papers using the search terms "Aeromonas+name of Arab country (i.e. Algeria, Egypt, etc.)". Additional data were obtained from a Google search using the aforementioned terms. The organisms have been reported from diarrheal children, patients with cholera-like diarrhea, an outbreak of acute gastroenteritis and from different types of animals, foods and water source in several Arab countries in the Middle East and North Africa with predominance of A. hydrophila, A. caviae and A. sobria. Using molecular techniques few studies reported genes encoding several toxins from aeromonads isolated from different sources. Among the antimicrobials examined in the present review third generation cephalosporins, fluoroquinolones and aminoglycosides showed excellent activity and can be employed in the treatment of Aeromonas-associated human infections in Arabic countries. Whenever possible, treatment should be guided by the susceptibility testing results of the isolated organism. In the future, studies employing molecular testing methods are required to provide data on circulating genospecies and their modes of transmission in the community, and on their mechanisms of resistance to antimicrobials. Microbiology laboratories and research centers are encouraged to look for these organisms in clinical, food and water sources to attain a better understanding of the public health risks from these organisms in Arab countries.

  19. Aeromonas spp. isolated from oysters (Crassostrea rhizophorea from a natural oyster bed, Ceará, Brazil Aeromonas spp. isoladas de ostras (Crassostrea rhizophorea coletadas em um criadouro natural, Ceará, Brazil

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    Norma S. Evangelista-Barreto

    2006-06-01

    Full Text Available Between April and October 2002, thirty fortnightly collections of oysters (Crassostrea rhizophorea from a natural oyster bed at the Cocó River estuary in the Sabiaguaba region (Fortaleza, Ceará, Brazil were carried out, aiming to isolate Aeromonas spp. strains. Oyster samples were submitted to the direct plating (DP and the presence/absence (P/A methods. Aeromonas were identified in 15 (50% samples analyzed by the DP method and in 13 (43% analyzed by the P/A method. A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii and Aeromonas sp. were isolated. The predominant species was A. veronii (both biovars, which was identified in 13 (43% samples, followed by A. media in 11 (37% and A. caviae in seven (23%. From the 59 strains identified, 28 (48% presented resistance to at least one of the eight antibiotics tested.Foram realizadas 30 coletas quinzenais, entre abril e outubro de 2002, de ostras (Crassostrea rhizophorea de um criadouro natural, no estuário do rio Cocó (Fortaleza/Ceará/Brasil, objetivando-se isolar cepas de Aeromonas spp. As amostras de ostras foram submetidas aos métodos de plaqueamento direto (PD e presença/ausência (P/A. Foram identificadas Aeromonas em 15 (50% amostras analisadas pelo método PD e em 13 (43% pelo método P/A. Foram isoladas: A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii e Aeromonas sp. A espécie predominate foi A. veronii (ambos biovars, identificada em 13 (43% amostras, seguida de A. media em 11 (37% e A. caviae em 7 (23%. Das 59 cepas identificadas, 28 (48% apresentaram resistência a pelo menos um, dos oitos antibióticos testados.

  20. Pneumonia caused by Aeromonas species in Taiwan, 2004-2011.

    Science.gov (United States)

    Chao, C M; Lai, C C; Tsai, H Y; Wu, C J; Tang, H J; Ko, W C; Hsueh, P-R

    2013-08-01

    We investigated the clinical characteristics of patients with pneumonia caused by Aeromonas species. Patients with pneumonia caused by Aeromonas species during the period 2004 to 2011 were identified from a computerized database of a regional hospital in southern Taiwan. The medical records of these patients were retrospectively reviewed. Of the 84 patients with pneumonia due to Aeromonas species, possible Aeromonas pneumonia was diagnosed in 58 patients, probable Aeromonas pneumonia was diagnosed in 18 patients, and pneumonia due to Aeromonas was conclusively diagnosed in 8 patients. Most of the cases of Aeromonas pneumonia developed in men and in patients of advanced age. A. hydrophila (n = 50, 59.5 %) was the most common pathogen, followed by A. caviae (n = 24, 28.6 %), A. veronii biovar sobria (n = 7, 8.3 %), and A. veronii biovar veronii (n = 3, 3.6 %). Cancer (n = 37, 44.0 %) was the most common underlying disease, followed by diabetes mellitus (n = 27, 32.1 %). Drowning-associated pneumonia developed in 6 (7.1 %) patients. Of 47 patients who were admitted to the intensive care ward, 42 patients developed acute respiratory failure and 24 of those patients died. The overall in-hospital mortality rate was significantly associated with liver cirrhosis, cancer, initial presentation of shock, and usage of mechanical ventilation. In conclusion, Aeromonas species should be considered as one of the causative pathogens of severe pneumonia, especially in immunocompromised patients, and should be recognized as a cause of drowning-associated pneumonia. Cirrhosis, cancer, and shock as the initial presenting symptom are associated with poor outcome.

  1. Ocorrência de bactérias do gênero Aeromonas em queijo-de-minas frescal artesanal Occurrence of the genus Aeromonas in minas frescal cheese

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    C.C.C. Bulhões

    2002-06-01

    Full Text Available Foram analisadas 160 amostras de queijo-de-minas frescal artesanal, adquiridas no comércio varejista dos municípios de Poços de Caldas - MG e Jaboticabal - SP, a fim de verificar a ocorrência de bactérias do gênero Aeromonas no produto. Oitenta e duas (51,2% encontravam-se contaminadas pelos microrganismos, com populações que variavam de 5,0×10³ a 4,0×10(5 UFC/grama. Foram identificadas as espécies Aeromonas hydrophila, Aeromonas caviae, Aeromonas schubertii, além de cepas consideradas atípicas. Os resultados evidenciam que bactérias do gênero Aeromonas podem ser veiculadas através do queijo tipo minas frescal artesanal e devem servir de alerta aos serviços de saúde pública.One hundred and sixty samples of homemade minas frescal cheese, obtained at the retail trade in the municipalities of Poços de Caldas-MG and Jaboticabal-SP, were analyzed for occurrence of the genus Aeromonas. Eighty two samples (51.2% were contaminated by these microorganisms with populations that ranged from 5.0 x 10³ to 4.0 x 10(5 CFU/g. The species Aeromonas hydrophila, Aeromonas caviae, Aeromonas schubertii, were identified, along with atypical strains. The results show that Aeromonas bacteria genus can be transmitted through the homemade minas frescal cheese and they should serve of alert to the public health services.

  2. Phylogenetic diversity of Aeromonas from "alheira," a traditional Portuguese meat product.

    Science.gov (United States)

    Fontes, M C; Martins, C; Martínez-Murcia, A J; Saavedra, M J

    2012-08-01

    "Alheira" is a traditional smoked meat sausage produced in the north of Portugal, representing an important economic resource for the region. This meat product has been subjected to research studies with the aim of detecting the presence of common foodborne pathogens, but, to our knowledge, isolation of emerging foodborne Aeromonas from alheira has never been previously described. Present work attempts to evaluate the Aeromonas species diversity of 84 isolates of Aeromonas spp. collected from 32 alheira samples. All presumptive Aeromonas isolates were subjected to genotyping by enterobacterial repetitive intergenic consensus-polymerase chain reaction analysis. The isolates presenting a different pattern were subjected to gyrB gene sequencing for species classification, and the species A. hydrophila, A. salmonicida, A. caviae, A. media, and A. allosaccharophila were identified. The Aeromonas species diversity found has not been previously described in any other meat product evaluated in previous studies. It is also important to highlight the presence of A. hydrophila and A. caviae because they were previously associated with illness in humans, including gastroenteritis.

  3. Bactérias do gênero Aeromonas em abatedouro de frangos Contamination by Aeromonas spp. in poultry slaughterhouse

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    F.N. Costa

    2002-10-01

    Full Text Available Analisaram-se 200 amostras de diferentes produtos e locais do fluxograma de abate de frangos com o objetivo de identificar os pontos de contaminação da carne de frango por Aeromonas. Das 25 amostras analisadas para cada um dos oito pontos estudados foram isoladas Aeromonas spp. em nove (36% amostras de penas, em 14 (56% de fezes, em 18 (72% de carcaças não evisceradas, evisceradas e resfriadas e em 20 (80% de água do pré-resfriamento. Não foram isoladas Aeromonas spp. da água de abastecimento da indústria e da água do tanque de escaldagem. Foram identificadas as espécies, Aeromonas hydrophila em 39 (15,2% amostras, A. sobria em 69 (26,9%, A. caviae em 87 (34%, A. veronii em 18 (7%, A. schubertii em três (1,2%, A. trota em duas (0,8% e A. jandaei em uma (0,4%. O resultado sugere que independente do controle higiênico-sanitário adotado na indústria, as carcaças de frangos podem se contaminar já a partir de sua obtenção, determinando o aparecimento de Aeromonas em carcaças resfriadas e prontas para a comercialização.Two hundred samples of poultry meat were analyzed in order to evaluate contamination by Aeromonas spp. in eight sites of a slaughterhouse, with 25 samples obtained in each site. Aeromonas spp. was found in nine (36.0% samples of feathers, 14 (56.0% samples of stools, 18 (72.0% carcasses after pre-evisceration, post-evisceration, and cooling, as well as in 20 (80.0% samples of chilling water. The microorganism was not found in samples from either the supplying water or the tank for scalding water. Aeromonahydrophila was identified in 39 (15.2% samples, whereas A. sobria, A. caviae, A. veronii, A. schubertii, A. trota, and A. jandaei were identified in, respectively, 69 (26.9%, 87 (34.0%, 18 (7.0%, three (1.2%, two (0.8%, and one (0.4% samples. In addition, bacteria dissemination may occur during slaughtering and regardless the hygienic-sanitary control followed by the industry, poultry carcasses may be contaminated

  4. Aeromonas dhakensis, an increasingly recognized human pathogen

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    Po Lin eChen

    2016-05-01

    Full Text Available Aeromonas dhakensis was first isolated from children with diarrhea in Dhaka, Bangladesh and described in 2002. In the past decade, increasing evidence indicate this species is widely distributed in the environment and can cause a variety of infections both in human and animals, especially in coastal areas. A. dhakensis is often misidentified as A. hydrophila, A. veronii or A. caviae by commercial phenotypic tests in the clinical laboratory. Correct identification relies on molecular methods. Increasingly used matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS may be able to identify Aeromonas specie rapidly and accurately. A. dhakensis has shown its potent virulence in different animal models and clinical infections. Although several virulence factors had been reported, no single mechanism is conclusive. Characteristically A. dhakensis is the principal species causing soft tissue infection and bacteremia, especially among patients with liver cirrhosis or malignancy. Of note, A. dhakensis bacteremia is more lethal than bacteremia due to other Aeromonas species. The role of this species in gastroenteritis remains controversial. Third generation cephalosporins and carbapenems should be used cautiously in the treatment of severe A. dhakensis infection due to the presence of AmpC β-lactamase and metallo-β-lactamase genes, and optimal regimens may be cefepime or fluoroquinolones. Studies of bacterial virulence factors and associated host responses may provide the chance to understand the heterogeneous virulence between species. The hypothesis A. dhakensis with varied geographic prevalence and enhanced virulence that compared to other Aeromonas species warrants more investigations.

  5. Aeromonas species isolated from PINTADO fish (Pseudoplatystoma sp: virulence factors and drug susceptibility

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    Vera Lúcia Mores Rall

    1998-09-01

    Full Text Available Aeromonas has been described as an emergent foodborne pathogen of increasing importance. In this study, we report that 48% of 50 Pintado fish samples collected at the retail market of São Paulo city were positive for Aeromonas sp, as detected by the direct plating method. When the presence/absence method was used, the positivity was 42%. A. caviae was the most frequent species, followed by A. hydrophila and A. sobria. Production of cytotoxic enterotoxin, observed in suckling mouse assay, was detected in 67% of A. sobria strains, in 60% of A. hydrophila strains and in 40% of A. caviae strains. In vitro tests, performed with HEp-2 cells, showed that 88% of A. hydrophila, 27% of A. sobria and 13% of A. caviae strains were positive for this toxin. The in vivo production of cytotonic enterotoxin, tested after heating the filtrates at 56ºC for 20 minutes, was detected in 17% of A. sobria, in 10% of A. caviae and in none of A. hydrophila strains in vivo. All analyzed strains did not alter HEp-2 cells. 20% and 16% of A. sobria and A. caviae isolates, respectively, presented capacity to adhere to HEp-2 cells. In counterpart, invasion of HEp-2 cells was not observed in any isolate. The Aeromonas isolates were sensitive to the majority of the antimicrobiol agents tested.Bactérias do gênero Aeromonas têm sido descritas como patógenos emergentes de importância crescente em alimentos. Neste estudo, relatamos que 48% das amostras de peixe "Pintado" coletado no comércio de São Paulo, foram positivas para Aeromonas sp quando isoladas pelo método de plaqueamento direto. Quando o método Presença/Ausência foi utilizado, a porcentagem de positividade foi de 42%. A. caviae foi a espécie mais freqüente, seguida por A. hydrophila e A. sobria. Produção de enterotoxina citotóxica, determinada em camundongos recém-nascidos, foi observada em 67% das cepas de A. sobria, em 60% das de A. hydrophila e em 40% das de A. caviae. No teste in vitro em c

  6. AEROMONAS SPP BACTEREMIA OF RAINBOW TROUT FRY (ONCORHYNCHUS MYKISS: BIOCHEMICAL CHARACTERIZATION OF THE CAUSATIVE ORGANISM AND ANTIMICROBIAL SUSCEPTIBILITY

    Directory of Open Access Journals (Sweden)

    Damir Kapetanović

    2004-09-01

    Full Text Available Aeromonas hydrophila and other members of Aeromonas genus are ubiquitus in aquatic environment and make part of normal bacterial flora of rainbow trout. Aeromonas spp. infections are worldwide registered. Here we present our experience and knowledge on Aeromonas bacteremia, which causes mortality of rainbow trout fry. Rainbow trout fry, 7 month old, started to die in November 2003. Fish samples (17 samples of dead and moribund fish were delivered to the Laboratory for aquaculture. With Api 20 NE tests Aeromonas hydrophila / caviae type I was identified with an average probability of 99.9 % (one test against, as well as Aeromonas hydrophila / caviae type II with an average probability of 99.5 % (one test against from liver, spleen, kidney, intestines and damaged eye. All of isolated and identified samples were tested for antibiotic susceptibility by disc diffusion method. The test showed that specimens were most sensitive on flumequin, and relatively less sensitive on chloramphenicol and enrofloxacin. Therapy was successfully applied with Flubactin®.

  7. Virulence and antimicrobial susceptibility of clinical and environmental strains of Aeromonas spp. from northeastern Brazil.

    Science.gov (United States)

    Castelo-Branco, Débora de Souza Collares Maia; Guedes, Glaucia Morgana de Melo; Brilhante, Raimunda Sâmia Nogueira; Rocha, Marcos Fábio Gadelha; Sidrim, José Júlio Costa; Moreira, José Luciano Bezerra; Cordeiro, Rossana de Aguiar; Sales, Jamille Alencar; Riello, Giovanna Barbosa; de Alencar, Lucas Pereira; Paiva, Manoel de Araújo Neto; Vasconcelos, David Caldas; de Menezes, Isis Sousa Bezerra; de Ponte, Yago Brito; Sampaio, Célia Maria de Souza; Monteiro, André Jalles; Bandeira, Tereza de Jesus Pinheiro Gomes

    2015-08-01

    The aims of the present study were to isolate and identify clinical and environmental strains of Aeromonas spp. by means of biochemical tests and the automated method VITEK 2 and to investigate the presence of the virulence genes cytotoxic enterotoxin (act), hemolysin (asa-1), and type III secretion system (ascV), and also the in vitro antimicrobial susceptibility of the strains. From the clinical isolates, 19 Aeromonas hydrophila, 3 Aeromonas veronii bv. sobria, and 1 Aeromonas caviae were identified, while from the environmental strains, 11 A. hydrophila, 22 A. veronii bv. sobria, 1 A. veronii bv. veronii, and 1 A. caviae were recovered. The gene act was detected in 69.5% of clinical isolates, asa-1 in 8.6%, and ascV in 34.7%. In the environmental strains, the detection rates were 51.4%, 45.7%, and 54.2% for the genes act, asa-1, and ascV, respectively. Resistance to amoxicillin-clavulanate and piperacillin-tazobactam was observed in 15 and 3 clinical strains, respectively, and resistance to ceftazidime, meropenem, imipenem, ciprofloxacin, and trimethoprim-sulfamethoxazole was observed in 1 strain for each drug. Resistance to amoxicillin-clavulanate and piperacillin-tazobactam was detected in 17 and 1 environmental strain, respectively. Higher resistance percentages were observed in clinical strains, but environmental strains also showed this phenomenon and presented a higher detection rate of virulence genes. Thus, it is important to monitor the antimicrobial susceptibility and pathogenic potential of the environmental isolates.

  8. Molecular characterization of Aeromonas species isolated from farmed eels (Anguilla japonica).

    Science.gov (United States)

    Yi, Seung-Won; You, Myung-Jo; Cho, Ho-Seong; Lee, Chang-Seop; Kwon, Joong-Ki; Shin, Gee-Wook

    2013-05-31

    Seventy Aeromonas strains were identified by phylogenetic analysis using housekeeping genes (gyrB and rpoD) in order to investigate etiological agents for aeromoniasis in farmed eels (Anguilla japonica). The phylogenetic analysis showed that Aeromonas aquariorum (n=22, 31.4%) was the predominant species among the investigated eel strains, followed by Aeromonas caviae (n=16, 22.9%), A. veronii (n=13, 18.6%), A. hydrophila (n=12, 17.1%), A. jandaei (n=4, 5.7%), A. media (n=2, 2.9%), and A. trota (n=1, 1.4%). The potential virulence of the present strains was estimated by performing PCR assays using the following seven virulence genes: cytotoxic enterotoxin (act), two cytotonic enterotoxins (alt and ast), glycerophospholipid:cholesterol acyltransferase (gcaT), DNase (exu), lipase (lip), and flagellin (fla). The detection rates of act, alt, ast, gcaT, exu, lip, and fla among all 70 strains were 91.4%, 55.7%, 27.1%, 97.1%, 95.7%, 100%, and 98.6%, respectively. In genotyping of enterotoxin genes, act(+)/alt(+)/ast(+), act(+)/alt(+)/ast(-), and act(+)/alt(-)/ast(-) genotypes were prevalent in A. hydrophila (8/12 strains), A. aquariorum (13/22 strains), and A. caviae (14/16 strains), respectively, suggesting a high heterogeneity among Aeromonas species. In this study, A. aquariorum, which has been an unrecorded species in Korea, can be an etiological agent for aeromoniasis of eel.

  9. Population dynamics and antimicrobial susceptibility of Aeromonas spp. along a salinity gradient in an urban estuary in Northeastern Brazil.

    Science.gov (United States)

    Silva, Camila Magalhães; Evangelista-Barreto, Norma Suely; Vieira, Regine Helena Silva Dos Fernandes; Mendonça, Kamila Vieira; de Sousa, Oscarina Viana

    2014-12-15

    The main objective of this study was to quantify population and identify culturable species of Aeromonas in sediment and surface water collected along a salinity gradient in an urban estuary in Northeastern Brazil. Thirty sediment samples and 30 water samples were collected from 3 sampling locations (A, B and C) between October 2007 and April 2008. The Aeromonas count was 10-7050CFU/mL (A), 25-38,500CFU/mL (B) andAeromonas caviae, A. sobria, A. trota, A. salmonicida and A. allosaccharophila) were identified among 41 isolates. All strains were sensitive to chloramphenicol and ceftriaxone, whereas 33 (80, 4%) strains were resistant to at least 2 of the 9 antibiotics tested. Resistance to erythromycin was mostly plasmidial. In conclusion, due to pollution, the Cocó River is contaminated by pathogenic strains of Aeromonas spp. with a high incidence of antibacterial resistance, posing a serious risk to human health.

  10. AQU-1, a chromosomal class C β-lactamase, among clinical Aeromonas dhakensis isolates: distribution and clinical significance.

    Science.gov (United States)

    Wu, Chi-Jung; Wang, Hsuan-Chen; Chen, Po-Lin; Chang, Ming-Chung; Sunny Sun, H; Chou, Pei-Hsin; Ko, Wen-Chien

    2013-11-01

    Aeromonas dhakensis, a recently described Aeromonas sp. formerly called Aeromonas aquariorum, is associated with human infections. In this study, a chromosomal gene, blaAQU-1, was identified in A. dhakensis AAK1 that constitutes a 1143-bp open reading frame and is 87% identical to the gene encoding CepH in Aeromonas hydrophila. An Escherichia coli TOP10 cell transformant harbouring blaAQU-1 was resistant to cefotaxime but not to cefepime. mRNA expression of blaAQU-1 in the cefotaxime-resistant mutant strain AAK1m was 70-fold higher than in the wild strain AAK1. In all 16 A. dhakensis isolates (the major species of 51 consecutive Aeromonas blood isolates collected from June 1999 to June 2001) as well as in A. aquariorum MDC47(T) and A. hydrophila subsp. dhakensis LMG 19562(T), but not in the reference strains or clinical isolates of other A. hydrophila subspecies, Aeromonas caviae, Aeromonas veronii or Aeromonas enteropelogenes, blaAQU-1-related genes were detected by PCR. Overall, 13 (81%) of the 16 A. dhakensis blood isolates exhibited either cefotaxime resistance or the in vitro emergence of derepressed cefotaxime-resistant mutants. In vivo selection of an A. dhakensis resistant mutant was noted in a burn patient undergoing cefotaxime monotherapy. These observations suggest that AQU-1 is a chromosomal cephalosporinase in A. dhakensis. Cefotaxime monotherapy for severe A. dhakensis infections should be used cautiously.

  11. Envolvimento de Aeromonas em surto de doença diarréica aguda em São Bento do Una, Pernambuco Aeromonas associated with an acute diarrhea outbreak in São Bento do Una, Pernambuco

    Directory of Open Access Journals (Sweden)

    Ernesto Hofer

    2006-04-01

    Full Text Available No primeiro semestre de 2004, ocorreu um surto de diarréia em São Bento do Una, Pernambuco, registrando-se 2.170 casos. Nas 582 coproculturas realizadas, 145 (25% revelaram um enteropatógeno bacteriano, destacando 114 casos (19,5% com a participação de Aeromonas, representadas por Aeromonas caviae (57/9,8%, Aeromonas veronii biovar sobria (23/3,9%, Aeromonas veronii biovar veronii (15/2,6% e outras espécies (19/3,2%. Nos 31 episódios restantes (5,3%, foram detectados: V. cholerae O1 Ogawa toxigênico (18/3,1%, Salmonella spp (8/1,4%, Shigella spp (3/0,5% e Vibrio cholerae não O1/não O139 (2/0,3%.An acute diarrhea outbreak, with 2170 cases, was described during January to July, 2004, in São Bento do Una, Pernambuco. 582 stools were examined and an enteric pathogen was recovered in 25% (145 patients. Aeromonas species were the most frequent (114-19.5% and the main isolates were Aeromonas caviae (57-9.8%, Aeromonas veronii biovar sobria (23-3.9%, Aeromonas veronii biovar veronii (15-2.6% and other species (19-3.2%. The other isolated enteropathogens were Vibrio cholerae O1-Ogawa toxigenic (18-3.1%, Salmonella spp (8-1.4%, Shigella spp (3-0.5% and Vibrio cholerae non-O1/non-O139 (2-0.3%.

  12. Prevalence of fur mites (Chirodiscoides caviae) in pet guinea pigs (Cavia porcellus) in southern Italy.

    Science.gov (United States)

    d'Ovidio, Dario; Santoro, Domenico

    2014-04-01

    Chirodiscoides caviae is the most common fur mite affecting guinea pigs; infestation is generally asymptomatic. No studies have been published on the prevalence of such mites in guinea pigs in southern Italy. We sought to evaluate the prevalence and the clinical signs of C. caviae infestation in guinea pigs in southern Italy. Clinical records of guinea pigs evaluated from August 2012 to July 2013 were retrospectively searched. In this retrospective matched case-control study, records of guinea pigs with evidence of C. caviae infestation were selected. The prevalence of C. caviae infestation was evaluated and exposure variables were assessed among guinea pigs with and without infestation using stepwise conditional logistic regression. Guinea pigs seen during the same time period, but without a diagnosis of C. caviae, were included as control animals. The prevalence of C. caviae was 32% (42 of 131); 66.6% of affected guinea pigs (28 of 42) originated from pet shops, whereas 28% (14 of 42) were privately owned. Thirty-one guinea pigs (73.8%) were asymptomatic, whereas 11 (26.1%) showed clinical signs (pruritus, alopecia, erythema and scaling). The most frequently affected area was the lumbosacral region (38 of 42). Guinea pigs in pet shops were more likely to be affected by C. caviae than owned guinea pigs (odds ratio, 5.12; 95% confidence interval, 2.32-11.29; P < 0.0001). The results of this study indicate a high prevalence of C. caviae infestation in guinea pigs in southern Italy. Chirodiscoides mites should be sought in guinea pigs, particularly in animals coming from pet shops. © 2014 ESVD and ACVD.

  13. The immunocross-reaction between pathogenic Aeromonas hydrophila isolated from Siberian sturgeon Acipenser baerii and other three Aeromonas strains%西伯利亚鲟嗜水气单胞菌与其它3株气单胞菌间的免疫交叉反应

    Institute of Scientific and Technical Information of China (English)

    姜有声; 杨倩; 杨先乐; 吕利群; 邱军强

    2012-01-01

    The immunological characteristics of four Aeromonas strains like pathogenic bacteria strain Xl(Aeromonas hydrophlia) isolated from Siberian sturgeon (Acipenser baerii) suffering with bacterial septicaemia, pathogenic bacteria strain XL2-T strain (Aeromonas caviae ) isolated from the sturgeon septicemia, pathogenic strain Wl (Aeromonas sobria) and non-pathogenic M3 strain (Aeromonas hydrophlia) , were detected through the cross-reactions between four Aeromonas strains and antiserums by indirect immunofluoresence assay. The results showed that the pathogenic XI strain, XL2 -T strain and Wl strain had strong cross reactions, and weak cross reactions were found on non-pathogenic M3 strain with other three pathogenic strains.%采用间接免疫荧光技术分析了西伯利亚鲟细菌性败血症致病菌嗜水气单胞菌(Aeromonas hydrophlia)X1菌株、豚鼠气单胞菌(Aeromonas caviae)XL2-T菌株、致病性温和气单胞菌(Aeromonas sobria)W1菌株与无致病性嗜水气单胞菌(Aeromonas hydrophlia)M3菌株等水产养殖主要病原菌与抗血清之间的免疫交叉反应.结果显示具有致病性的同属菌株X1菌株、XL2-T菌株、W1菌株交叉反应程度较大,说明这3株菌表面存在较多相同抗原决定簇.而无致病性菌株M3与其他3株致病性菌株免疫交叉反应程度较小.

  14. Aeromonas species in stabilization ponds in the arid region of Marrakesh, Morocco, and relation to fecal-pollution and climatic factors.

    Science.gov (United States)

    Boussaid, A; Baleux, B; Hassani, L; Lesne, J

    1991-12-01

    During the period 12 July 1985 to 23 December 1987, water samples were collected in two-week intervals for estimates ofAeromonas species in a waste treatment system located in the arid region of Marrakech, Morocco. Fecal coliforms, temperature, and chemical oxygen demand were measured simultaneously withAeromonas species densities. Statistical methods were utilized to analyze the significance of average differences and temporal patterns ofAeromonas species numbers.Removal ofAeromonas in the whole system did not exceed 1.14 log.Aeromonas densities showed significantly higher resistance to the treatment process when compared with fecal coliforms; however, abundance of the two groups presented a similar seasonal change. The highest numbers occurred during the cold months, while the lowest appeared in the warm months. Statistical time-series analyses of the densities data showed the seasonal and cyclic distribution ofAeromonas in this treatment plant.These temporal changes were simultaneously observed in all the stations investigated and were negatively correlated with water temperature values.Aeromonas populations were dominated byA. caviae andA. hydrophila in the inlet samples. These two species were rapidly eliminated in the treatment plant. The temporal distribution ofA. caviae was similar to the change in densities ofAeromonas and fecal coliforms. The seasonal fluctuations of abundance ofAeromonas were probably related to this species, which dominated in the winter samples but dropped during the summer. Meanwhile,A. sobria dominated all the final effluent samples. This greater survival ofA. sobria and its known pathogenicity may limit the re-use of treated water for irrigation of fodder plants.

  15. Molecular detection of the Aeromonas virulence aerolysin gene in retail meats from different animal sources in Egypt.

    Science.gov (United States)

    Osman, Kamelia; Aly, Magdy; Kheader, Afaf; Mabrok, Khaled

    2012-05-01

    Meat commonly contain the same Aeromonas spp. which occur in human diarrhoeal and non-diarrhoeal faecal samples. Motile Aeromonas were isolated from 5.6% of total 302 samples. The distribution of the isolates were 5.9 and 5.2% in fresh and frozen samples, respectively. Of the 302 samples taken of the four animal meat species investigated, the genus Aeromonas were isolated in 12.3% of the fresh samples collected from buffalo meat, in 6.5% of the samples collected from sheep meat and 14.0% from the samples collected from the cattle frozen meat samples. The camel meat did not reveal any Aeromonas isolates. Aeromonas hydrophila was isolated as the most prevalent species with 6.8%, followed by Aeromonas caviae with 2.7% and Aeromonas sobria with 2.1% from the total meat samples. Aerolysin toxin gene (aerA) was detected in 3/17 isolates of A. hydrophila isolated from contaminated meat. Infection due to bacterial pathogen with such virulent factor through contact with contaminated meat while handling them, poses health hazards to humans.

  16. Multidrug-resistant (MDR) Aeromonas recovered from the metropolitan area of Valencia (Spain): diseases spectrum and prevalence in the environment.

    Science.gov (United States)

    Esteve, C; Alcaide, E; Giménez, M J

    2015-01-01

    Aeromonas infections are rare in Europe and often related to traveller's diarrhoea. A total of 185 Aeromonas isolates from river water, fish and clinical sources, recovered during a 1-year period, were used to investigate the disease spectrum and impact of multidrug-resistant (MDR) strains. They were all identified by biochemical tests and 25% of them were also identified by sequencing of the 16S rRNA gene. The minimum inhibitory concentrations (MICs) of 21 antimicrobials were determined for all isolates by broth microdilution/E-strips methods, and susceptibility was assessed according to the Clinical and Laboratory Standards Institute (CLSI). Strains pathogenicity was determined by using Swiss Webster mice as the animal model. Aeromonas diseases had an incidence of around 20 cases/million inhabitants in the metropolitan area of Valencia (Spain). Acute gastroenteritis in children with no history of travel abroad was the main pathology. These cases were related to A. caviae, A. veronii biovar sobria, A. hydrophila and A. dhakensis. A significant incidence of A. caviae in humans was found, while the other species were equally present in clinical and environmental origins. A. jandaei, A. bestiarum and A. media had mainly an environmental distribution. The prevalence of MDR Aeromonas was maximal in clinical samples, and resistance phenotypes were significantly related to this source. 7.2% of environmental Aeromonas was resistant to at least five drugs; most of them were moderately virulent for mice and, in addition, belonged to clinically significant species. The present study demonstrates a diseases spectrum similar to that reported in tropical countries, and also that pathogenic and heavily MDR Aeromonas are present in environmental reservoirs. MDR Aeromonas from any source analysed were susceptible to aztreonam, netilmicin, cefotaxime, ceftazidime, cefepime and fluoroquinolones.

  17. A molecular study on the prevalence and virulence potential of Aeromonas spp. recovered from patients suffering from diarrhea in Israel.

    Directory of Open Access Journals (Sweden)

    Yigal Senderovich

    Full Text Available BACKGROUND: Species of the genus Aeromonas are native inhabitants of aquatic environments and have recently been considered emerging human pathogens. Although the gastrointestinal tract is by far the most common anatomic site from which aeromonads are recovered, their role as etiologic agents of bacterial diarrhea is still disputed. Aeromonas-associated diarrhea is a phenomenon occurring worldwide; however, the exact prevalence of Aeromonas infections on a global scale is unknown. METHODOLOGY/PRINCIPAL FINDINGS: The prevalence and virulence potential of Aeromonas in patients suffering from diarrhea in Israel was studied using molecular methods. 1,033 diarrheal stools were sampled between April and September 2010 and Aeromonas species were identified in 17 (∼2% patients by sequencing the rpoD gene. Aeromonas species identity and abundance was: A. caviae (65%, A. veronii (29% and Aeromonas taiwanensis (6%. This is the first clinical record of A. taiwanensis as a diarrheal causative since its recent discovery from a wound infection in a patient in Taiwan. Most of the patients (77% from which Aeromonas species were isolated were negative for any other pathogens. The patients ranged from 1 to 92 years in age. Aeromonas isolates were found to possess different virulence-associated genes: ahpB (88%, pla/lip/lipH3/apl-1 (71%, act/hlyA/aerA (35%, alt (18%, ast (6%, fla (65%, lafA (41%, TTSS ascV (12%, TTSS ascF-ascG (12%, TTSS-dependent ADP-ribosylating toxins aexU (41% and aexT (6% in various combinations. Most of the identified strains were resistant to beta-lactam antibiotics but susceptible to third-generation cephalosporin antibiotics. CONCLUSIONS: Aeromonas may be a causative agent of diarrhea in patients in Israel and therefore should be included in routine bacteriological screenings.

  18. SELEKSI KANDIDAT PROBIOTIK ANTI AEROMONAS HYDROPHILA UNTUK PENGENDALIAN PENYAKIT IKAN AIR TAWAR

    Directory of Open Access Journals (Sweden)

    Angela Mariana Lusiastuti

    2012-08-01

    Full Text Available Secara dasar ada tiga model kerja probiotik yaitu menekan populasi mikroba melalui kompetisi dengan memproduksi senyawa anti-mikroba atau melalui kompetisi nutrisi dan tempat pelekatan di dinding usus, mengubah keseimbangan metabolisme mikroba dengan meningkatkan atau menurunkan aktivitas enzim dan menstimulasi imunitas dengan meningkatkan antibodi dan aktivitas makrofag. Tujuan penelitian ini adalah memperoleh mikroba yang berpotensi sebagai kandidat probiotik untuk dapat diaplikasikan di dalam menekan atau menghambat bakteri patogen sebagai langkah pengendalian penyakit pada ikan air tawar. Penelitian ini terdiri atas dua tahap, tahap pertama adalah tahap seleksi yang dimulai dari koleksi dan isolasi bakteri kandidat probiotik dan tahap kedua adalah tahap pengujian bakteri kandidat probiotik secara in vitro menggunakan metode daya hambat dan menguji daya patogenisitas bakteri. Total isolat bakteri yang diperoleh adalah 72 isolat, dan hanya enam isolat terpilih sebagai kandidat probiotik anti Aeromonas hydrophila. Probiotik anti A. hydrophila adalah sebagai berikut: Chromobacterium lividum, Alcaligenes faecalis, Aeromonas caviae, Bacteriodes serpens, Bacillus firmus, Aeromonas caviae = A. hydrophila Sub sp. Anaerogenes.

  19. Taxonomía y epidemiología del género aeromonas

    OpenAIRE

    Alperi Vega, Anabel

    2009-01-01

    Durante la presente tesis doctoral se ha establecido la presencia de variabilidad interoperónica en el gen ARNr 16S de Aeromonas y observado que ésta afectaba a la taxonomía del género, limitando la identificación de A. caviae, A. media y A. veronii. La secuenciación del gen rpoD permitió identificar las cepas con variabilidad interoperónica a nivel de especie y reconocer 5 nuevas especies del género: A. fluvialis, A. taiwanensis, A. sanarelii, A. piscicola y A. rivuli. En colaboración con d...

  20. Prevalence and diversity of Aeromonas and Vibrio spp. in coastal waters of Southern Italy

    DEFF Research Database (Denmark)

    Dumontet, S.; Krovacek, K.; Svenson, S.B.

    2000-01-01

    and A. caviae. Furthermore, the antibiotic resistance patterns of isolated bacteria showed the presence of a number of resistant strains among the isolates. In order to discriminate the isolates on the basis of their biochemical profiles and/or antibiotic resistance patterns, cluster analysis...... was carried out which showed that no unique assay could fully discern these isolates. However, the best discrimination resulted from complete pattern profile based on both biochemical profiles and antibiotic resistance patterns.......% of samples were positive for Vibrio spp. It was interesting to note that 38% of the positive stations for both Aeromonas and Vibrio spp. showed a fecal coliform contamination of water at bacterial indicators (such as fecal...

  1. Occurrence, molecular characterization, and antimicrobial susceptibility of Aeromonas spp. in marine species of shrimps cultured at inland low salinity ponds.

    Science.gov (United States)

    Yano, Yutaka; Hamano, Kaoru; Tsutsui, Isao; Aue-Umneoy, Dusit; Ban, Masatoshi; Satomi, Masataka

    2015-05-01

    We aimed to document the risk of Aeromonas spp. in marine shrimp species cultured in inland low salinity ponds in Thailand. In 14 of 18 shrimp samples retrieved from inland grow-up ponds, Aeromonas spp. were detected at ranges from 4667 to 1,500,000 CFU/g body weight. The phylogenetic tree constructed with the gyrB and cpn60 concatenated sequences indicated that the 87 isolates consisted of Aeromonas veronii (70%), Aeromonas aquariorum (18%), Aeromonas caviae (7%), Aeromonas jandaei (2%), and Aeromonas schubertii (2%). The potential virulence of the isolates was examined by phenotypic and PCR assays. Hemolytic activity and the extracellular activity of lipase, DNase, and gelatinase were observed in most isolates (94-99%). PCR revealed the presence of 9 genes related to virulence in the 87 isolates: act (75%), aer (74%), alt (30%), ast (1%), ascV (34%), aexT (24%), fla (92%), ela (34%), and lip (24%). The susceptibility profiles to 14 antimicrobial agents of isolates were typical for the genus, but resistance to cefotaxime, a third-generation cephalosporin, and imipenem were found in two A. aquariorum and in three A. veronii isolates, respectively. These resistances were confirmed by determining minimum inhibitory concentrations. Our results indicate that the microbiological risk posed by Aeromonas should be considered for marine shrimp species that are cultured in low-salinity ponds. These shrimps may also be a vehicle for the transfer of different genotypes of Aeromonas and antibiotic-resistant determinants to regions worldwide through trade.

  2. Occurrence of motile Aeromonas in municipal drinking water and distribution of genes encoding virulence factors.

    Science.gov (United States)

    Pablos, Manuel; Rodríguez-Calleja, Jose M; Santos, Jesús A; Otero, Andrés; García-López, María-Luisa

    2009-10-31

    Aeromonas-associated cases of gastroenteritis are generally considered waterborne. The purpose of this study was to evaluate the potential microbiological risk associated with the presence of these bacteria in public drinking water. Over a period of one year, 132 drinking-water samples were monitored in León (NW of Spain, 137,000 inhabitants) for mandatory drinking-water standards and the occurrence of Aeromonas spp. Samples were taken at the municipal water treatment plant, one storage facility, and two public artesian drinking-water fountains. Because of low numbers of coliforms or Clostridium perfringens, the non-compliance rate with microbial standards was 3.8% whereas the percentage of positive samples for motile mesophilic Aeromonas was 26.5%. For all but two samples, Aeromonas was recovered between October and early March when the temperature was below 14 degrees C and the residual chlorine ranged from 0.21 to 0.72 mg/l. An apparent relationship was observed between rainfall and the incidence of Aeromonas. The 35 selected Aeromonas isolates were identified as A. caviae and A. media. The alt and laf genes were present in all isolates, the aerA gene was present in six isolates, and the four remaining genes investigated (hlyA, ast, stx1 and stx2) were absent. The combinations of putative virulence genes were: aerA(-)/hlyA(-)/alt(+)/ast(-)/laf(+)/stx1(-)/stx2(-) (82.9%) and aerA(+)/hlyA(-)/alt(+)/ast(-)/laf(+)/stx1(-)/stx2(-) (17.1%). None of the isolates bore plasmids. As Aeromonas strains harbouring two or more virulence-associated genes have the potential to cause disease by direct transmission via drinking water or by water use in food preparation, it would be advisable to control excessive numbers of these bacteria in drinking-water supplies.

  3. Chlorinated and ultraviolet radiation -treated reclaimed irrigation water is the source of Aeromonas found in vegetables used for human consumption.

    Science.gov (United States)

    Latif-Eugenín, Fadua; Beaz-Hidalgo, Roxana; Silvera-Simón, Carolina; Fernandez-Cassi, Xavi; Figueras, María J

    2017-04-01

    Wastewater is increasingly being recognized as a key water resource, and reclaimed water (or treated wastewater) is used for irrigating vegetables destined for human consumption. The aim of the present study was to determine the diversity and prevalence of Aeromonas spp. both in reclaimed water used for irrigation and in the three types of vegetables irrigated with that water. Seven of the 11 (63.6%) samples of reclaimed water and all samples of vegetables were positive for the presence of Aeromonas. A total of 216 Aeromonas isolates were genotyped and corresponded to 132 different strains that after identification by sequencing the rpoD gene belonged to 10 different species. The prevalence of the species varied depending on the type of sample. In the secondary treated reclaimed water A. caviae and A. media dominated (91.4%) while A. salmonicida, A. media, A. allosaccharophila and A. popoffii represented 74.0% of the strains in the irrigation water. In vegetables, A. caviae (75.0%) was the most common species, among which a strain isolated from lettuce had the same genotype (ERIC pattern) as a strain recovered from the irrigation water. Furthermore, the same genotype of the species A. sanarellii was recovered from parsley and tomatoes demonstrating that irrigation water was the source of contamination and confirming the risk for public health. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Molecular characterization of Aeromonas spp. and Vibrio cholerae O1 isolated during a diarrhea outbreak

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    Carina Lucena Mendes-Marques

    2012-12-01

    Full Text Available This work aimed to assess pathogenic potential and clonal relatedness of Aeromonas sp. and Vibrio cholerae isolates recovered during a diarrhea outbreak in Brazil. Clinical and environmental isolates were investigated for the presence of known pathogenic genes and clonal relatedness was assessed by intergenic spacer region (ISR 16S-23S amplification. Four Aeromonas genes (lip, exu, gcat, flaA/B were found at high overall frequency in both clinical and environmental isolates although the lip gene was specifically absent from selected species. A fifth gene, aerA, was rarely found in A. caviae, the most abundant species. The ISR profile revealed high heterogeneity among the Aeromonas isolates and no correlation with species identification. In contrast, in all the V. cholerae isolates the four genes investigated (ctxA, tcpA, zot and ace were amplified and revealed homogeneous ISR and RAPD profiles. Although Aeromonas isolates were the major enteric pathogen recovered, their ISR profiles are not compatible with a unique cause for the diarrhea events, while the clonal relationship clearly implicates V. cholerae in those cases from which it was isolated. These results reinforce the need for a better definition of the role of aeromonads in diarrhea and whether they benefit from co-infection with V. cholerae.

  5. CHLORINE DISINFECTION OF AEROMONAS

    Science.gov (United States)

    The bacterial genus Aeromonas is currently listed on the USEPA's Candidate Contaminant List (CCL). Resistance to chemical disinfection is an essential aspect regarding all microbial groups listed on the CCL. This study was designed to determine the inactivation kinetics of Aeromo...

  6. Distribution of 13 virulence genes among clinical and environmental Aeromonas spp. in Western Australia.

    Science.gov (United States)

    Aravena-Román, M; Inglis, T J J; Riley, T V; Chang, B J

    2014-11-01

    We evaluated the pathogenic potential of 98 clinical and 31 environmental Aeromonas isolates by detecting the presence of 13 virulence genes using a polymerase chain reaction (PCR)-based method. The majority (96 %) of the strains contained at least one of the virulence genes. The overall distribution was aerA/haem (77 %), alt (53 %), lafA (51 %), ast (39 %), flaA (32 %), aspA (29 %), vasH (26 %), ascV (16 %) and aexT (13 %). No amplification products were detected for the genes encoding a bundle-forming pilus (BfpA and BfpG) or a Shiga-like toxin (stx-1 and stx-2). Five or more virulence genes were detected in 42 % of environmental and 24 % of clinical isolates. Among the major species, 48 % of A. hydrophila and 42 % of A. dhakensis isolates harboured five or more virulence genes compared with 19 % in A. veronii bv. sobria and none in A. caviae isolates. Our results suggest that, in Western Australia, strains of A. dhakensis and A. hydrophila are potentially more virulent than those of A. veronii bv. sobria and A. caviae, although the pathogenic potential of Aeromonas spp. is probably strain- rather than species-dependent.

  7. Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia.

    Science.gov (United States)

    Khor, Wei Ching; Puah, Suat Moi; Tan, Jin Ai Mary Anne; Puthucheary, S D; Chua, Kek Heng

    2015-01-01

    Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.

  8. Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia.

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    Wei Ching Khor

    Full Text Available Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT and RNA polymerase sigma-factor (rpoD genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.

  9. Molecular identification of 4 clinical Aeromonas isolates by gyrB and rpoB gene sequencing%4株气单胞菌临床分离株基于gyrB和rpoB基因序列的分子鉴定

    Institute of Scientific and Technical Information of China (English)

    韦小瑜; 游旅; 田克诚; 朱琳; 张仁俊; 吉光辉; 唐光鹏; 王定明

    2016-01-01

    目的:对4株气单胞菌临床分离株进行菌种的分子鉴定,为气单胞菌的种型鉴定提供病原学依据。方法对4株系统生化鉴定为气单胞菌的菌株,提取核酸作为模板,分别使用gyrB、rpoB基因进行扩增及产物测序,将所得序列在GenBank中进行同源性搜索,与GenBank中已经公布的同源性较高的气单胞菌序列比较构建系统发育树并进行分析。结果4株气单胞菌gyrB基因序列经同源性搜索,与多株嗜水气单胞菌和豚鼠气单胞菌同源性为99%,rpoB基因序列则与多株豚鼠气胞菌同源性为99%~100%,对gyrB基因和rpoB基因序列分别进行系统发育树分析显示,4株气单胞菌菌株在gyrB基因系统发育树中与多株嗜水气单胞菌和豚鼠气单胞菌聚为一簇,而在rpoB基因系统发育树中与多株豚鼠气单胞菌聚在同一分枝内。结论4株气单胞菌经rpoB基因测序分析明确鉴定为豚鼠气单胞菌,gyrB基因测序分析不能有效鉴定其种型。%Objective To identify the gene species of 4 clinical Aeromonas isolates and to provide the etiologic basis for the identification of Aeromonas species. Methods The isolates were identified as Aeromonas by conventional biochemical methods. The DNA of Aeromonas isolates was extracted as the template of PCR. The gyrB and rpoB gene were amplified and sequenced. Two sequence homology searches were conducted on GenBank and selected closely homology strains. Two molecular phylogenetic trees were constructed for analyze. Results The sequence homology of gyrB gene was 99% among Aeromonas isolates、 Aeromonas hydrophila and Aeromonas caviae strains,99%~100%(rpoB) among Aeromonas isolates and Aeromonas caviae strains. The phylogenetic tree of gyrB gene showed that 4 Aeromonas isolates、Aeromonas caviae strains and Aeromonas hydrophila strains were clustered in the same group. While 4 Aeromonas isolates fell into the same branch with Aeromonas caviae strains

  10. Efficacy of doramectin in Trixacarus caviae infestation in guinea pigs (Cavia porcellus).

    Science.gov (United States)

    Singh, Shanker K; Dimri, Umesh; Ahmed, Quazi Shahir; Sayedda, Kauser; Singh, Krishna Veer

    2013-04-01

    The present study was intended to evaluate the efficacy of doramectin against seven naturally Trixacarus caviae infested male guinea pigs. Multiple skin scrapings of all the seven guinea pigs were found microscopically positive for T. caviae mites. Clinically these animals revealed, more or less denuded, very red often thickened, and crustated cutaneous lesions restricted at the sacral region and back. Doramectin 1 % (w/v) was administered intramuscularly at a dose rate of 400 μg/kg once weekly, which resulted in profound improvements in clinical conditions within 14 days after the first doramectin application. It took almost 28 days for the cutaneous lesions to disappear and to witness partial hair coat regrowth. Two moderately infested guinea pigs required only single injection of doramectin to achieve complete parasitological cure, while remaining five (one moderately infested and four severely infested) guinea pigs required two injections of doramectin to achieve complete parasitological cure. No adverse effects were revealed by any of the doramectin treated guinea pigs during the study period. Thus, it can be concluded from the present study that guinea pigs naturally infested by T. caviae mites can be cured safely using two doses of doramectin once in a week.

  11. Two membrane filter media (mADA/0129 and mSA/0129 agars) for enumeration of motile Aeromonas in seawater.

    Science.gov (United States)

    Alonso, J L; Garay, E

    1989-10-01

    Sewage-contaminated natural seawater was analysed for the presence of motile Aeromonas by two membrane filtration procedures. Incubation of membranes on two modified media (mADA (0/129) and mSA (0/129) were compared. The specificity of the two media was high, 95.8% on mADA (0/129) and 94.8% on mSA (0129). The most frequent species identified were A. caviae, followed by A. hydrophila and A. sobria. Motile Aeromonas counts were high in both media (greater than 10(4)/100 ml) and no significant differences were observed between them. The two membrane filtration procedures allowed rapid quantitative recovery of motile Aeromonas from seawater in the presence of very large numbers of competing microflora.

  12. Occurrence of potential pathogenic Aeromonas species in tropical seafood, aquafarms and mangroves off Cochin coast in South India

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    Alphonsa Vijaya Joseph

    2013-12-01

    Full Text Available Background: The genus Aeromonas include gram-negative, motile, facultative anaerobic, rod shaped and oxidase positive bacteria comprising several species, associated with the aquatic environment. Aeromonas species have been implicated in human pathogenesis and are linked with gastroenteritis, muscle infections, septicemia, and skin diseases. In fish they are renowned as enteric pathogens causing haemorrhagic septicemia, fin rot, soft tissue rot and furunculosis resulting in major die-offs and fish kills. Aim: This study reports the occurrence of potential pathogenic Aeromonas sp. in tropical seafood (Squids, Prawns and Mussels, aquafarms and mangroves of Cochin, Kerala, South India. Materials and Methods :Tropical seafood (Squid, Prawn and Mussel, sediment and water samples from aquafarms and associated mangroves were screened for Aeromonas contamination. The isolates were identified by 16S rDNA sequence analysis and subjected to morphological and biochemical characterization. Haemolytic assay was used for determining pathogenicity of the organisms. Antibiotic susceptibility against 12 antibiotics were performed and the MAR index was calculated. Results: A total of 134 isolates were recovered from the samples of which 15 were identified as Aeromonas species by 16S rDNA sequence analysis and were assigned to 5 species namely, A. hydrophila, A. enteropelogenes, A. caviae, A. punctataand A. aquarorium. Morphological, biochemical and phylogenetic analyses revealed relatedness and variability among the strains. All the isolates were haemolytic on blood agar indicating their pathogenicity. The isolates exhibited varying degrees of resistance to vancomycin (86.66%, ampicillin (46.66%, nalidixic acid (20%, tetracycline (6.66%, co-trimaxozole (6.66% and rifampicin (6.66% and were susceptible to antibiotics like gentamycin, streptomycin, trimethoprim, azithromycin, cefixime and chloramphenicol. 20% of Aeromonas sp. showed MAR index > 0.2 indicative of the

  13. Aeromonas associated diarrhoeal disease in south Brazil: prevalence, virulence factors and antimicrobial resistance Aeromonas associadas a diarréias no sul do Brasil: prevalência, fatores de virulência, e resistência a antibiótico

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    Ivani M.F. Guerra

    2007-12-01

    Full Text Available Aeromonas were isolated from 27 (6.6% of 408 patients admitted with acute gastroenteritis in two hospitals at Rio Grande do Sul, Brazil. Isolates were classified as A. hydrophila (51.8%, A. caviae (40.8%, and A. veronii biotype sobria (7.4%. The highest prevalence of Aeromonas associated infections occurred in lactants and children. Virulence genes (aerA -aerolysin/hemolysin, ahpA -serine-protease, satA - glycerophospholipid-cholesterol acyltransferase, lipA -lipase, and ahyB -elastase and virulence factors (hemolytic, proteolitic, lipolitic activities, and biofilm formation were identified in most A. hydrophila and A. veronii biotype sobria isolates, with lower frequencies on A. caviae. All Aeromonas isolates were resistant to ampicillin, ticarcillin/clavulanic acid, cephalotin, and cephazolin, and most of them (>70% exhibited resistance to imipenem, carbenicillin, amoxillin/sulbactan, and piperacillin. Multiple-resistance, more than four antibiotics, was evidenced in 29.6% of the isolates. The most efficient antibiotics were the quinolones (ciprofloxacin and norfloxacin, and the aminoglycosides (amikacin and netilmicin.Aeromonas foram isoladas de 27 (6.6% dos 408 pacientes admitidos com gastroenterite aguda em dois hospitais do Rio Grande do Sul, Brasil. Os isolados foram classificados com A. hydrophila (51.8%, A. caviae (40.8%, e A. veronii biotype sobria (7.4%. A maior prevalência de Aeromonas ocorreu em lactantes e crianças. Genes (aerA -aerolisina/hemolisina, ahpA -serina-protease, satA - glicerofosfolipidio-colesterol aciltransferase, lipA -lipase, e ahyB -elastase e factores (atividade hemolítica, proteolítica, lipolítica, e formação de biofilme de virulência foram identificados na maioria dos isolados de A. hydrophila e A. veronii biotype sobria, com freqüências menores em A. caviae. Todos os isolados de Aeromonas apresentaram resistência a ampicilina, ticarcilina/ácido clavulânico, cefalotina e cefazolina, e a maior parte

  14. Biological control of Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) using Aeromonas phage PAS-1.

    Science.gov (United States)

    Kim, J H; Choresca, C H; Shin, S P; Han, J E; Jun, J W; Park, S C

    2015-02-01

    The potential control efficacy of Aeromonas phage PAS-1 was evaluated against Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) model in this study. The phage was co-cultured with the virulent A. salmonicida subsp. salmonicida strain AS05 that possesses the type III secretion system (TTSS) ascV gene, and efficient bacteriolytic activity was observed against the bacteria. The administration of PAS-1 in rainbow trout demonstrated that the phage was cleared from the fish within 200 h post-administration, and a temporal neutralizing activity against the phage was detected in the sera of phage-administrated fish. The administration of PAS-1 (multiplicity of infection: 10 000) in A. salmonicida subsp. salmonicida infected rainbow trout model showed notable protective effects, with increased survival rates and mean times to death. These results demonstrated that Aeromonas phage PAS-1 could be considered as an alternative biological control agent against A. salmonicida subsp. salmonicida infections in rainbow trout culture.

  15. Clinical manifestations of bacteremia caused by Aeromonas species in southern Taiwan.

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    Hung-Jen Tang

    Full Text Available This study is conducted to investigate the clinical characteristics of patients with bacteremia caused by Aeromonas species.Patients with bacteremia caused by Aeromonas species during the period 2009 to 2013 were identified from a computerized database of a regional hospital in southern Taiwan. The medical records of these patients were retrospectively reviewed.A total of 91 patients with bacteremia due to Aeromonas species were identified. In addition to 16 (17.6% primary bacteremia, the most common source of secondary infection is peritonitis (n = 27, 29.7%, followed by biliary tract infection (n = 18, 19.8%, and SSTI (n = 12, 13.2%, pneumonia (n = 9, 9.9%, catheter-related bloodstream infection (n =  5, 5.5%, and genitourinary tract infection (n = 4, 4.4%. A. hydrophila (n = 35, 38.5% was the most common pathogen, followed by A. veronii biovar sobria (n = 31, 34.1%, A. caviae (n = 14, 15.4%, and A. veronii biovar veronii (n = 9, 9.9%. Forty-three (47.3% patients were classified as healthcare-associated infections (HCAI causes by Aeromonas species, and patients with HCAI were more likely to have cancer, and receive immunosuppressant than patients with community-acquired bacteremia. The overall outcomes, including rate of ICU admission, acute respiratory failure, and mortality were 33.3%, 28.6%, and 23.1%, respectively. Multivariate analysis showed that the in-hospital day mortality was significantly associated only with underlying cancer (P <.001, and initial shock (P <.001.Aeromonas species should be considered one of the causative pathogens of healthcare-associated bacteremia, especially in immunocompromised patients. In addition, it can be associated with high fatality. Cancer and initial shock were the poor prognostic factors.

  16. Aeromonas spp.: an emerging pathogen?

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    Andrea Bartolini

    2015-12-01

    Full Text Available The aim of this study is to identify and monitor the presence of Aeromonas spp. strains in stool cultures. We analyzed 5564 stool cultures from September 2012 to August 2013. Sixty-three patients were positive for Aeromonas spp. The most frequent symptoms were: diarrhea (46.0% and abdominal pain (12.7%. Pediatric subjects were 28. Samples’ microscopic examination showed leukocytes in 38.1% of cases. It is still controversial whether Aeromonas are responsible for human gastroenteritis, but their presence in faecies of symptomatic patients supports their etiologic role. We propose search for toxins by polymerase chain reaction to identify strains that require an antibiotic therapy.

  17. Quinolone resistant Aeromonas spp. as carriers and potential tracers of acquired antibiotic resistance in hospital and municipal wastewater.

    Science.gov (United States)

    Varela, Ana Rita; Nunes, Olga C; Manaia, Célia M

    2016-01-15

    Members of the genus Aeromonas are recognized carriers of antibiotic resistance in aquatic environments. However, their importance on the spread of resistance from hospital effluents to the environment is poorly understood. Quinolone resistant Aeromonas spp. (n = 112) isolated from hospital effluent (HE) and from raw (RWW) and treated wastewater (TWW) of the receiving urban wastewater treatment plant (UWTP) were characterized. Species identification and genetic intraspecies diversity were assessed based on the 16S rRNA, cpn60 and gyrB genes sequence analysis. The antibiotic resistance phenotypes and genotypes (qnrA, qnrB, qnrC, qnrD, qnrS, qnrVC; qepA; oqxAB; aac(6′)-Ib-cr; blaOXA; incU) were analyzed in function of the origin and taxonomic group. Most isolates belonged to the species Aeromonas caviae and Aeromonas hydrophila (50% and 41%, respectively). The quinolone and the beta-lactamase resistance genes aac(6′)-Ib-cr and blaOXA, including gene blaOXA-101, identified for the first time in Aeromonas spp., were detected in 58% and 56% of the isolates, respectively, with identical prevalence in HE and UWTP wastewater. In contrast, the gene qnrS2 was observed mainly in isolates from the UWTP (51%) and rarely in HE isolates (3%), suggesting that its origin is not the clinical setting. Bacterial groups and genes that allow the identification of major routes of antibiotic resistance dissemination are valuable tools to control this problem. In this study, it was concluded that members of the genus Aeromonas harboring the genes aac(6′)-Ib-cr and blaOXA are relevant tracers of antibiotic resistance dissemination in wastewater habitats, while those yielding the gene qnrS2 allow the traceability from non-clinical sources.

  18. Characterization of Aeromonas strains isolated from Indian foods using rpoD gene sequencing and whole cell protein analysis.

    Science.gov (United States)

    Nagar, Vandan; Shashidhar, Ravindranath; Bandekar, Jayant R

    2013-04-01

    Aeromonas are responsible for causing gastroenteritis and extra-intestinal infections in humans. Twenty-two Aeromonas strains isolated from different food sources were re-identified up to species level using rpoD gene sequence analysis. Biochemical tests and 16S rRNA gene sequencing were insufficient to identify Aeromonas till species level. However, incorporation of additional biochemical tests lead to correct identification of 95.5 % strains up to species level. The 16S rRNA gene sequencing was useful to identify Aeromonas isolates at the genus level only. Sequences of the rpoD gene showed greater discriminatory power than 16S rRNA gene and provided conclusive discrimination of the strains for which the phenotypic species identification was uncertain. All these 22 strains were accurately identified up to species level by rpoD gene as A. salmonicida (6), A. veronii bv. veronii (4), A. caviae (3), A. hydrophila (2), A. veronii bv. sobria (2), A. jandaei (1), A. trota (1), A. sobria (1), A. allosaccharophila (1) and A. bivalvium (1). All these strains were also characterized using whole cell protein (WCP) analysis by gradient SDS-PAGE and showed different whole cell protein (WCP) profile [22-28 polypeptide bands (~10 to >97 kDa)], indicating high genetic diversity. The present work emphasizes the use of molecular methods such as rpoD gene sequencing along with comprehensive biochemical tests for the rapid and accurate identification of Aeromonas isolates till species level. The WCP profile can be subsequently used to characterize Aeromonas isolates below species level.

  19. Biochemical characteristics, serogroups, and virulence factors of aeromonas species isolated from cases of diarrhoea and domestic water samples in Chennai

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    Alavandi S

    2003-01-01

    Full Text Available PURPOSE: The objective of the present study was to delineate the differences between the clinical and environmental Aeromonas species with respect to their biochemical characteristics, serogrouping and virulence factors, in order to find a phenotypic marker of enteropathogenicity. METHODS: A total of 55 Aeromonas spp. inclusive of 19 isolates from cases of diarrhoea, and 36 from water samples comprising, 10 isolates of A. hydrophila, 21 isolates each of A. sobria, and A. caviae, two isolates of A. jandaei and one isolate of A. veronii were subjected to analysis of their biochemical characteristics, serogrouping, and virulence factors. RESULTS: Among the differences recorded in the biochemical characteristics in the three major species, the most striking characteristic was fermentation of lactose, which was observed in all the 11 A. caviae isolates recovered from water samples. None of the 10 clinical isolates of A. caviae tested fermented lactose. The clinical Aeromonas isolates belonged to seven typable serogroups, O:13, O:14, O:16, O:21, O:27, O:32 and O:35. The environmental isolates belonged to eight different serogroups, such as, O:3, O:11, O:14, O:16, O:18, O:28, O:64 and O:78 and were predominated by serotypes O:18 and O:64. Among the virulence factors tested, 89% of the environmental isolates produced b haemolysin, while only 62.3% of clinical isolates were able to do so. There was no significant difference between the clinical and environmental aeromonads with respect to their enterotoxigenicity in suckling mice in vivo, cytotoxicity in vitro in Vero cell monolayers, and ability to produce siderophores. CONCLUSION: Efforts to delineate the differences between the clinical and environmental Aeromonas spp. did not reveal significant difference between them. However, difference was observed with respect to their ability to produce b haemolysin, wherein, higher percentage of environmental isolates was haemolytic. The results also suggest

  20. Streptococcus caviae sp. nov., isolated from Guinea pig faecal samples

    NARCIS (Netherlands)

    Palakawong Na Ayudthaya, Susakul; Hilderink, Loes J.; Oost, van der John; Vos, de Willem M.; Plugge, Caroline M.

    2017-01-01

    A novel cellobiose-degrading and lactate-producing bacterium, strain Cavy grass 6T, was isolated from faecal samples of guinea pigs (Cavia porcellus). Cells of the strain were ovalshaped, non-motile, non-spore-forming, Gram-stain-positive and facultatively anaerobic. The strain gr at 2

  1. Leishmaniose cutânea em cobaias (Cavia porcellus Cutaneous leishmaniasis in guinea pigs (Cavia porcellus

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    Roselene Ecco

    2000-06-01

    Full Text Available São descritos três casos de leishmaniose cutânea em cobaias (Cavia porcellus machos de dois anos de idade que eram criadas como animais de estimação. As lesões macroscópicas eram semelhantes em duas cobaias e consistiam de nódulos cutâneos ulcerados no aspecto dorsal do pavilhão auricular de ambas as orelhas. Na outra cobaia, a lesão consistia de engrossamentos irregulares na pele do escroto que apareceram após uma orquiectomia realizada dois meses antes. A histopatologia era semelhante nos três casos e consistia de acentuada dermatite granulomatosa, com ulceração e numerosas formas protozoárias amastigotas no citoplasma de macrófagos, sugestivas de Leishmania sp. A microscopia eletrônica confirmou o diagnóstico de leishmaniose.Three cases of cutaneous leishmaniasis in three two-year-old male guinea pigs (Cavia porcellus raised as pets are described. Gross lesions were similar in two of the guinea pigs and consisted of raised and ulcerated cutaneous nodules in the dorsal aspect of the pinna of both ears. In another guinea pig the lesion consisted of irregular thickenning in the skin of the scrotum which followed an orchiectomy performed two months before. Histopathology was similar in the three cases and consisted of severe diffuse granulomatous dermatitis with ulceration and numerous intrahistiocytic protozoal amastigotes resembling Leishmania sp. Electron microscopy confirmed the diagnosis of leishmaniasis.

  2. Influence of food system conditions on N-acyl-L-homoserine lactones production by Aeromonas spp.

    Science.gov (United States)

    Medina-Martínez, M S; Uyttendaele, M; Demolder, V; Debevere, J

    2006-12-01

    Eleven of 13 Aeromonas strains were shown to produce AHLs. Results of TLC showed that N-butanoyl-L-homoserine lactone (C4-HSL) was the main AHL produced in LB medium at 30 degrees C. The influence of different carbon sources, temperature, pH values and salt concentrations on AHL production was determined in eight A. hydrophila and one A. caviae strain. Additionally a quantitative study of C4-HSL production by A. hydrophila strain 519 under different conditions was performed. Positive results were found in the AHL induction assay for some Aeromonas strains in cultures in LB agar incubated at 12 degrees C after 72-96 h. The induction of the sensor strains by Aeromonas spp. occurred in LB medium supplemented with all carbon sources in a concentration of 0.5%. The production of C4-HSL by A. hydrophila 519 was found until 3.5% (w/v) of NaCl. For pHs close to the neutrality the C4-HSL production by A. hydrophila was evident after 24-48 h of incubation. A. hydrophila 519 produced C4-HSL under anaerobic conditions. Also, the AHL production by Aeromonas strains was studied in simulate agar of shrimp, fish and some vegetables. The production of AHLs was evident by almost all the test strains in shrimp simulated agar. In fish agar only for one of three fish species tested, positive results were found. Induction assay in vegetables simulated agar showed principally negative results, probably because of the presence of inhibitory compounds in these vegetables.

  3. Distribution and phenotypic and genotypic detection of a metallo-β-lactamase, CphA, among bacteraemic Aeromonas isolates.

    Science.gov (United States)

    Wu, Chi-Jung; Chen, Po-Lin; Wu, Jiunn-Jong; Yan, Jing-Jou; Lee, Chin-Chi; Lee, Hsin-Chun; Lee, Nan-Yao; Chang, Chia-Ming; Lin, Yu-Tzu; Chiu, Yen-Cheng; Ko, Wen-Chien

    2012-05-01

    The objectives of the study were to investigate the distribution of cphA-related genes (cphA) encoding a CphA metallo-β-lactamase (MBL) among 51 consecutive Aeromonas blood isolates and to compare different phenotypic methods for detecting CphA. The presence of cphA was detected by PCR. Four phenotypic methods, the imipenem-EDTA combined disc test, imipenem-EDTA MBL Etest, agar dilution test and modified Hodge test (MHT), were used to detect imipenem susceptibility and MBL production. The results showed that 35 (69%) blood isolates had cphA. All (100%) of 16 Aeromonas aquariorum isolates and 12 Aeromonas veronii isolates, and 4 (80%) of 5 Aeromonas hydrophila isolates, carried cphA, but none of 15 Aeromonas caviae isolates did. With the standard inocula, irrespective of the presence or absence of cphA, all but one (50, 98%) isolates were susceptible to imipenem tested by disc diffusion, Etest and agar dilution (10(4) c.f.u. spot inocula), and did not exhibit MBL production by the imipenem-EDTA combined disc test and MBL Etest. By the agar dilution test using large inocula (10(7) c.f.u.), 34 (97%) of 35 cphA(+) isolates had imipenem MICs of ≥16 µg ml(-1), higher than the susceptible breakpoint (4 µg ml(-1)), and demonstrated positive results for the MHT, while one cphA(+) and all 17 cphA(-) isolates had imipenem MICs of ≤4 µg ml(-1). In conclusion, the distribution of cphA among aeromonads is species-specific, found in A. aquariorum, A. veronii and A. hydrophila, and the MHT may be a phenotypic screening test for CphA production.

  4. Mycetoma caused by Nocardia caviae in the first Brazilian patient.

    Science.gov (United States)

    Magalhães, Geraldo Magela; Oliveira, Sílvia Cristina de Castro; Soares, Ana Cláudia de Brito; Machado-Pinto, Jackson; de Resende, Maria Aparecida

    2010-01-01

    Mycetoma is a chronic subcutaneous mycosis caused by exogenous fungi or actinomycetes. This infection has a progressive course and shows a typical clinical characteristic of tumefaction, draining sinuses, and grains. Infection initiation is related to local trauma and can spread to muscle, underlying bone, and adjacent organs. Nocardia brasiliensis is the most frequent actinomycete isolated, while N. caviae is a rare agent. We present a case of mycetoma in a 37-year-old African-American man on the right hand. The infection had been apparent for four years prior to the consultation. When the infection did not respond to antibiotic therapy, the patient was referred to the Dermatology department. Routine laboratory studies were normal. X-ray examination of the hand showed an osteolytic lesion on the hand bones. On skin biopsy culture, on Sabouraud Dextrose Agar at 28 degrees C, a colony was isolated which was further identified as N. caviae by biochemical and hydrolysis testing. The patient was treated with oral trimethoprim/sulfamethoxazole (TMP/SMZ) 160/800 mg twice a day for 10 months. Four months after the beginning of the therapy, the subject exhibited clinical improvement and functional recovery of the hand. Five-year follow-up X-ray examination of the hand showed no osteolytic lesion on the hand bones. We report the first mycetoma case caused by N. caviae in our country with an unusual location on the hand. The patient showed clinical improvement with oral TMP/SMZ.

  5. Functional genomic characterization of virulence factors from necrotizing fasciitis-causing strains of Aeromonas hydrophila.

    Science.gov (United States)

    Grim, Christopher J; Kozlova, Elena V; Ponnusamy, Duraisamy; Fitts, Eric C; Sha, Jian; Kirtley, Michelle L; van Lier, Christina J; Tiner, Bethany L; Erova, Tatiana E; Joseph, Sandeep J; Read, Timothy D; Shak, Joshua R; Joseph, Sam W; Singletary, Ed; Felland, Tracy; Baze, Wallace B; Horneman, Amy J; Chopra, Ashok K

    2014-07-01

    The genomes of 10 Aeromonas isolates identified and designated Aeromonas hydrophila WI, Riv3, and NF1 to NF4; A. dhakensis SSU; A. jandaei Riv2; and A. caviae NM22 and NM33 were sequenced and annotated. Isolates NF1 to NF4 were from a patient with necrotizing fasciitis (NF). Two environmental isolates (Riv2 and -3) were from the river water from which the NF patient acquired the infection. While isolates NF2 to NF4 were clonal, NF1 was genetically distinct. Outside the conserved core genomes of these 10 isolates, several unique genomic features were identified. The most virulent strains possessed one of the following four virulence factors or a combination of them: cytotoxic enterotoxin, exotoxin A, and type 3 and 6 secretion system effectors AexU and Hcp. In a septicemic-mouse model, SSU, NF1, and Riv2 were the most virulent, while NF2 was moderately virulent. These data correlated with high motility and biofilm formation by the former three isolates. Conversely, in a mouse model of intramuscular infection, NF2 was much more virulent than NF1. Isolates NF2, SSU, and Riv2 disseminated in high numbers from the muscular tissue to the visceral organs of mice, while NF1 reached the liver and spleen in relatively lower numbers on the basis of colony counting and tracking of bioluminescent strains in real time by in vivo imaging. Histopathologically, degeneration of myofibers with significant infiltration of polymorphonuclear cells due to the highly virulent strains was noted. Functional genomic analysis provided data that allowed us to correlate the highly infectious nature of Aeromonas pathotypes belonging to several different species with virulence signatures and their potential ability to cause NF.

  6. Ribosomal multi-operon diversity: an original perspective on the genus Aeromonas.

    Directory of Open Access Journals (Sweden)

    Frédéric Roger

    Full Text Available 16S rRNA gene (rrs is considered of low taxonomic interest in the genus Aeromonas. Here, 195 Aeromonas strains belonging to populations structured by multilocus phylogeny were studied using an original approach that considered Ribosomal Multi-Operon Diversity. This approach associated pulsed-field gel electrophoresis (PFGE to assess rrn operon number and distribution across the chromosome and PCR-temporal temperature gel electrophoresis (TTGE to assess rrs V3 region heterogeneity. Aeromonads harbored 8 to 11 rrn operons, 10 operons being observed in more than 92% of the strains. Intraspecific variability was low or nul except for A. salmonicida and A. aquariorum suggesting that large chromosomic rearrangements might occur in these two species while being extremely rarely encountered in the evolution of other taxa. rrn operon number at 8 as well as PFGE patterns were shown valuable for taxonomic purpose allowing resolution of species complexes. PCR-TTGE revealed a high rate of strains (41.5% displaying intragenomic rrs heterogeneity. Strains isolated from human samples more frequently displayed intragenomic heterogeneity than strains recovered from non-human and environmental specimens. Intraspecific variability ranged from 0 to 76.5% of the strains. The observation of species-specific TTGE bands, the recovery of identical V3 regions in different species and the variability of intragenomic heterogeneity (1-13 divergent nucleotides supported the occurrence of mutations and horizontal transfer in aeromonad rrs evolution. Altogether, the presence of a high number of rrn operon, the high proportion of strains harboring divergent rrs V3 region and the previously demonstrated high level of genetic diversity argued in favor of highly adaptative capabilities of aeromonads. Outstanding features observed for A. caviae supported the ongoing process of adaptation to a specialized niche represented by the gut, previously hypothesized. 16S rRNA gene is an

  7. Prevalence, virulence and antimicrobial resistance patterns of Aeromonas spp. isolated from children with diarrhea

    Science.gov (United States)

    Soltan Dallal, Mohammad Mehdi; Mazaheri Nezhad Fard, Ramin; Kavan Talkhabi, Morteza; Aghaiyan, Leyla; Salehipour, Zohre

    2016-01-01

    Background Aeromonas spp. cause various intestinal and extraintestinal diseases. These bacteria are usually isolated from fecal samples, especially in children under five years old. The aim of this study was to assess the prevalence of Aeromonas spp. and their antimicrobial resistance profile in children with diarrhea referred to the Children Medical Center in Tehran, between 2013 and 2014. Methods A total number of 391 stool samples were collected from children with ages between 1 day and 14 years old, with diarrhea (acute or chronic), referred to the Children Hospital, Tehran, Iran, between 2013 and 2014. Samples were enriched in alkaline peptone water broth for 24 hours at 37 °C and then cultured. Suspicious colonies were analyzed through biochemical tests. Furthermore, antimicrobial susceptibility tests were carried out for the isolates. Isolates were further studied for act, ast, alt, aerA and hlyA virulence genes using polymerase chain reaction. Results In total, 12 isolates (3.1%) were identified as Aeromonas spp.; all were confirmed using the API-20E test. Of these isolates, five A. caviae (42%), four A. veronii (33%) and three A. hydrophila (25%) were identified in cases with gastroenteritis. Second to ampicillin (which was included in the growth medium used), the highest rate of antimicrobial resistance was seen against nalidixic acid and trimethoprim-sulfamethoxazole (5 isolates each, 41.6%) and the lowest rate of antimicrobial resistance was seen against gentamicin, amikacin and cefepime (none of the isolates). Results included 76.4% act, 64.7% ast, 71.5% alt, 83.3% aerA and 11.7% hlyA genes. Conclusion Aeromonas spp. are important due to their role in diarrhea in children; therefore, isolation and identification of these fecal pathogens should seriously be considered in medical laboratories. Since virulence genes play a significant role in gastroenteritis symptoms caused by these bacteria, Aeromonas species that include virulence genes are potentially

  8. Relation of chironomids with Aeromonas species

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    Sivan eLaviad

    2016-05-01

    Full Text Available Chironomids (Diptera: Chironomidae, also known as non-biting midges, are one of the most abundant groups of insects in aquatic habitats. They undergo a complete metamorphosis of four life stages of which three are aquatic (egg, larva, pupa, and the adult emerges into the air. Chironomids serve as a natural reservoir of Aeromonas and Vibrio cholerae species. Here we review existing knowledge about the mutual relations between Aeromonas species and chironomids. Using 454-pyrosequencing of the 16S rRNA gene, we found that the prevalence of Aeromonas species in the insects’ egg masses and larvae was 1.6% and 3.3% of the insects’ endogenous microbiota, respectively. Aeromonas abundance per egg mass remained stable during a six-month period of bacterial monitoring. Different Aeromonas species were isolated and some demonstrated the ability to degrade the insect’s egg masses and to prevent eggs hatching. Chitinase was identified as the enzyme responsible for the egg mass degradation. Different Aeromonas species isolated from chironomids demonstrated the potential to protect their host from toxic metals. Aeromonas is a causative agent of fish infections. Fish are frequently recorded as feeding on chironomids. Thus, fish might be infected with Aeromonas species via chironomid consumption. Aeromonas strains are also responsible for causing gastroenteritis and wound infections in humans. Different virulence genes were identified in Aeromonas species isolated from chironomids. Chironomids may infest drinking water reservoirs, hence be the source of pathogenic Aeromonas strains in drinking water. Chironomids and Aeromonas species have a complicated mutual relationship.

  9. Prevalence, Pathogenesis, Antibiotic Susceptibility Profiles, and In-vitro Activity of Selected Medicinal Plants Against Aeromonas Isolates from Stool Samples of Patients in the Venda Region of South Africa

    Science.gov (United States)

    Obi, C.L.; Ramalivhana, J.; Samie, A.; Igumbor, E.O.

    2007-01-01

    The prevalence, pathogenic indices, such as haemolytic and haemagglutinating activities, antibiograms, and in-vitro activities of local medicinal plants against Aeromonas isolates in Vhembe district of Limpopo province, South Africa, were studied using standard microbiological methods. In total, 309 diarrhoeic stool samples were collected from patients attending five health centres in the region during December 2004–May 2005. Aeromonas species were identified using the API 20E system. The haemagglutinating and haemolytic activities of isolates on human, sheep, pig and chicken red blood cells were investigated. Antibiotic susceptibility profiles of the isolates to several antibiotics and in-vitro activity of local medicinal plants were also ascertained using previously-reported schemes. Results showed that 104 (33.6%) of the 309 samples were positive for Aeromonas species, of which 89 (85.6%) were Aeromonas hydrophila, 12 (11.5%) A. sobria, and three (2.9%) A. caviae. All strains of A. hydrophila and A. caviae produced haemolysis on sheep blood, while eight of the 12 A. sobria strains were haemolytic on sheep blood. The haemolytic activities of the isolates were variable on other red blood cells tested. High level of resistance was observed to amoxicillin and ampicillin, followed by cefuroxime (79%), chloramphenicol (74%), and erythromycin (65%). The carbapenems were the most active drugs with only 7% resistance to meropenem and 11% to imipenem. About 12% of the isolates were resistant to ciprofloxacin. The extracts of three of seven medicinal plants tested showed inhibitory activity against all Aeromonas isolates; these included acetone and hexane extracts of Pterocarpus angolensis, Syzygium cordatum, and Zornia milneana. The results suggest a high prevalence of Aeromonas species in the region. The isolates demonstrated multiple resistant profiles to different antibiotics tested. Some local medicinal plants were inhibitory to Aeromonas isolates, indicating a

  10. [Aeromonas spp. and Plesiomonas shigelloides in bivalves, mud, and water from the Gulf of Nicoya, Costa Rica].

    Science.gov (United States)

    Rodríguez, E; Antillón, F

    1989-06-01

    Bivalves, mud, and surface water were collected at three different sites of the Gulf of Nicoya, Costa Rica, in search of Aeromonas spp. and Plesiomonas shigelloides. For their isolation, these bacteria were enriched in alkaline peptone water and streaked on MacConkey agar and on brilliant green bile inositol agar. This was followed by the biochemical tests necessary for their identification. Thirty-five strains of A. hydrophila, 58 of A. caviae, 43 of A. sobria, and 7 of P. shigelloides were isolated. None of these predominated nor was there any indication of a seasonal distribution along the 15 month's duration of the study. Seven strains of A. hydrophila and two of A. sobria showed the biochemical characteristics associated with toxin production (positive Voges-Proskauer and lysine decarboxylase tests). These species are widely distributed in the gulf and there is risk of contracting an infection while bathing or when eating raw bivalves from this area.

  11. Chlamydial development is blocked in host cells transfected with Chlamydophila caviae incA.

    Science.gov (United States)

    Alzhanov, Damir; Barnes, Jennifer; Hruby, Dennis E; Rockey, Daniel D

    2004-07-01

    Chlamydiae produce a set of proteins, termed Inc proteins, that are localized to the inclusion membrane and exposed to the host cell cytosol. Little information exists regarding the interaction of Inc proteins with the eukaryotic cell. To examine these interactions, Vaccinia virus vectors and mammalian plasmid-based systems were used to express inc genes in mammalian cells. Cells transfected with plasmids expressing Chlamydophila caviae incA were not productively infected by C. caviae. Expression of C. caviae incA also reduced inclusion formation by Chlamydia trachomatis, but not to the degree seen for C. caviae. Chlamydia trachomatis incA did not block development of either C. trachomatis or C. caviae. Deletion mutagenesis was used to demonstrate that plasmids encoding either the amino or carboxy-terminal regions of the protein, as well as the changing of a single amino acid within IncA (serine 17) could not block C. caviae infection. Immunoblot analysis of truncated IncA in a Vaccinia virus system provided evidence that serine 17 of C. caviae IncA is a target for phosphorylation. These experiments provide insight into the interaction of Inc proteins with the host cell and introduce a model system where these interactions can be explored further.

  12. Chlamydial development is blocked in host cells transfected with Chlamydophila caviae incA

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    Barnes Jennifer

    2004-07-01

    Full Text Available Abstract Background Chlamydiae produce a set of proteins, termed Inc proteins, that are localized to the inclusion membrane and exposed to the host cell cytosol. Little information exists regarding the interaction of Inc proteins with the eukaryotic cell. To examine these interactions, Vaccinia virus vectors and mammalian plasmid-based systems were used to express inc genes in mammalian cells. Results Cells transfected with plasmids expressing Chlamydophila caviae incA were not productively infected by C. caviae. Expression of C. caviae incA also reduced inclusion formation by Chlamydia trachomatis, but not to the degree seen for C. caviae. Chlamydia trachomatis incA did not block development of either C. trachomatis or C. caviae. Deletion mutagenesis was used to demonstrate that plasmids encoding either the amino or carboxy-terminal regions of the protein, as well as the changing of a single amino acid within IncA (serine 17 could not block C. caviae infection. Immunoblot analysis of truncated IncA in a Vaccinia virus system provided evidence that serine 17 of C. caviae IncA is a target for phosphorylation. Conclusions These experiments provide insight into the interaction of Inc proteins with the host cell and introduce a model system where these interactions can be explored further.

  13. [Application of cell co-culture techniques in medical studies].

    Science.gov (United States)

    Luo, Yun; Sun, Gui-Bo; Qin, Meng; Yao, Fan; Sun, Xiao-Bo

    2012-11-01

    As the cell co-culture techniques can better imitate an in vivo environment, it is helpful in observing the interactions among cells and between cells and the culture environment, exploring the effect mechanisms of drugs and their possible targets and filling the gaps between the mono-layer cell culture and the whole animal experiments. In recently years, they has attracted much more attention from the medical sector, and thus becoming one of research hotspots in drug research and development and bio-pharmaceutical fields. The cell co-culture techniques, including direct and indirect methods, are mainly used for studying pathological basis, new-type treatment methods and drug activity screening. Existing cell co-culture techniques are used for more pharmacological studies on single drug and less studies on interaction of combined drugs, such as collaborative compatibility and attenuation and synergistic effect among traditional Chinese medicines (TCMs). In line with the action characteristics of multi-component and multi-target, the cell co-culture techniques provide certain reference value for future studies on the effect and mechanism of combined TCMs on organisms as well as new methods for studies on TCMs and their compounds. This essay summarizes cell co-culture methods and their application and look into the future of their application in studies on TCMs and compounds.

  14. Virulent Aeromonas hydrophila in channel catfish

    Science.gov (United States)

    In this study, we investigated factors that predisposed catfish to motile aeromonas septicemia (MAS) caused by virulent Aeromonas hydrophila (vAh). Our results revealed that wounding on fish body surface was a prerequisite for vAh infection and disease development. A reproducible waterborne challeng...

  15. Aeromonas hydrophila and Aeromonas veronii predominate among potentially pathogenic ciprofloxacin- and tetracycline-resistant aeromonas isolates from Lake Erie.

    Science.gov (United States)

    Skwor, Troy; Shinko, Jasmine; Augustyniak, Alexander; Gee, Christopher; Andraso, Greg

    2014-02-01

    Members of the genus Aeromonas are ubiquitous in nature and have increasingly been implicated in numerous diseases of humans and other animal taxa. Although some species of aeromonads are human pathogens, their presence, density, and relative abundance are rarely considered in assessing water quality. The objectives of this study were to identify Aeromonas species within Lake Erie, determine their antibiotic resistance patterns, and assess their potential pathogenicity. Aeromonas strains were isolated from Lake Erie water by use of Aeromonas selective agar with and without tetracycline and ciprofloxacin. All isolates were analyzed for hemolytic ability and cytotoxicity against human epithelial cells and were identified to the species level by using 16S rRNA gene restriction fragment length polymorphisms and phylogenetic analysis based on gyrB gene sequences. A molecular virulence profile was identified for each isolate, using multiplex PCR analysis of six virulence genes. We demonstrated that Aeromonas comprised 16% of all culturable bacteria from Lake Erie. Among 119 Aeromonas isolates, six species were identified, though only two species (Aeromonas hydrophila and A. veronii) predominated among tetracycline- and ciprofloxacin-resistant isolates. Additionally, both of these species demonstrated pathogenic phenotypes in vitro. Virulence gene profiles demonstrated a high prevalence of aerolysin and serine protease genes among A. hydrophila and A. veronii isolates, a genetic profile which corresponded with pathogenic phenotypes. Together, our findings demonstrate increased antibiotic resistance among potentially pathogenic strains of aeromonads, illustrating an emerging potential health concern.

  16. The involvement of tetA and tetE tetracycline resistance genes in plasmid and chromosomal resistance of Aeromonas in Brazilian strains

    Directory of Open Access Journals (Sweden)

    Ilana Teruszkin Balassiano

    2007-11-01

    Full Text Available This study analyzed the involvement of tetA and tetE genes in the tetracycline resistance of 16 strains of genus Aeromonas, isolated from clinical and food sources. Polymerase chain reactions revealed that 37.5% of the samples were positive for tetA, and also 37.5% were tetE positive. One isolate was positive for both genes. Only the isolate A. caviae 5.2 had its resistance associated to the presence of a plasmid, pSS2. The molecular characterization of pSS2 involved the construction of its restriction map and the determination of its size. The digestion of pSS2 with HindIII originated two fragments (A and B that were cloned separately into the pUC18 vector. The tetA gene was shown to be located on the HindIII-A fragment by PCR. After transforming a tetracycline-sensitive strain with pSS2, the transformants expressed the resistance phenotype and harbored a plasmid whose size was identical to that of pSS2. The results confirmed the association between pSS2 and the tetracycline resistance phenotype, and suggest a feasible dissemination of tetA and tetE among strains of Aeromonas. This study suggests the spreading tetA and tetE genes in Aeromonas in Brazil and describes a resistance plasmid that probably contributes to the dissemination of the resistance.

  17. Microfluidics co-culture systems for studying tooth innervation

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    Pierfrancesco ePagella

    2014-08-01

    Full Text Available Innervation plays a key role in the development and homeostasis of organs and tissues of the orofacial complex. Among these structures, teeth are peculiar organs as they are not innervated until later stages of development. Furthermore, the implication of neurons in tooth initiation, morphogenesis and differentiation is still controversial. Co-cultures constitute a valuable method to investigate and manipulate the interactions of nerve fibres with their target organs in a controlled and isolated environment. Conventional co-cultures between neurons and their target tissues have already been performed, but these cultures do not offer optimal conditions that are closely mimicking the in vivo situation. Indeed, specific cell populations require different culture media in order to preserve their physiological properties. In this study we evaluate the usefulness of a microfluidics system for co-culturing mouse trigeminal ganglia and developing teeth. This device allows the application of specific media for the appropriate development of both neuronal and dental tissues. The results show that mouse trigeminal ganglia and teeth survive for long culture periods in this microfluidics system, and that teeth maintain the attractive or repulsive effect on trigeminal neurites that has been observed in vivo. Neurites are repealed when co-cultured with embryonic tooth germs, while postnatal teeth exert an attractive effect to trigeminal ganglia-derived neurons.In conclusion, microfluidics system devices provide a valuable tool for studying the behaviour of neurons during the development of orofacial tissues and organs, faithfully imitating the in vivo situation.

  18. Presumptive and Definitive Identification of Aeromonas from infected Ornamental Gold fish (Carassius auratus auratus)

    OpenAIRE

    2013-01-01

    The paper deals with the presumptive identification of Aeromonas, definitive identification of two species of Aeromonas i.e. Aeromonas hydrophila, Aeromonas salmonicida in various organs of Gold fish infected with red disease.

  19. Estimation of Value-at-Risk for Energy Commodities via CAViaR Model

    Science.gov (United States)

    Xiliang, Zhao; Xi, Zhu

    This paper uses the Conditional Autoregressive Value at Risk model (CAViaR) proposed by Engle and Manganelli (2004) to evaluate the value-at-risk for daily spot prices of Brent crude oil and West Texas Intermediate crude oil covering the period May 21th, 1987 to Novermber 18th, 2008. Then the accuracy of the estimates of CAViaR model, Normal-GARCH, and GED-GARCH was compared. The results show that all the methods do good job for the low confidence level (95%), and GED-GARCH is the best for spot WTI price, Normal-GARCH and Adaptive-CAViaR are the best for spot Brent price. However, for the high confidence level (99%), Normal-GARCH do a good job for spot WTI, GED-GARCH and four kind of CAViaR specifications do well for spot Brent price. Normal-GARCH does badly for spot Brent price. The result seems suggest that CAViaR do well as well as GED-GARCH since CAViaR directly model the quantile autoregression, but it does not outperform GED-GARCH although it does outperform Normal-GARCH.

  20. Helminths of the guinea pig, Cavia porcellus (Linnaeus, in Brazil

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    Roberto Magalhães Pinto

    2002-07-01

    Full Text Available Worm burdens were evaluated and compared in two groups of the guinea pig, Cavia poreellus (Linnaeus, 1758: animals of the first group were conventionally maintained in an institutional animal house and those of the second group were openly kept in pet shops in Brazil. Animals from both sources were infected only with the nematode Paraspidodera uncinata (Rudolphi, 1819 Travassos, 1914 (10% of prevalence in guinea pigs from lhe institutional facility and 40% in those animals from the pet shop. Other helminth samples recovered from Brazilian guinea pigs during 52 years and that are deposited in the Helminthological Collection of the Oswaldo Cruz Institute (CHIOC were also analyzed. Paraspidodera uncinata and the cestode Monoecocestus parcitesticulatus Rêgo, 1960 were identified in these samples.

  1. antimicrobial susceptibility pattern of aeromonas hydrophila among ...

    African Journals Online (AJOL)

    DR. AMINU

    immediately leaving the treatment plant may contain between 0-102 ... Diseases associated with Aeromonas are intestinal ... major symptoms (diarrhea, vomiting and fever) and duration of ... cholera (Ramamurthy et al., 1992), all identified A.

  2. PHENOTYPING, VIRULENCE CHARACTERISTICS OF AEROMONAS SPECIES AND THE EFFECTS OF ESSENTIAL PLANT OILS AS ANTIMICROBIAL AGENTS AGAINST PATHOGENIC ISOLATES FROM DIFFERENT SOURCES

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    Ahmed M.A. Mansour

    2014-01-01

    Full Text Available Aeromonas species are increasingly recognized as enteric pathogens. Faecal samples from 20 cow, 45 sheep; 60 goat and 60 camels were examined for the presence of Aeromonas species, which was also sought in the available drinking water (55 well water and 52 drinking chlorinated tap water were also examined. Aeromonas species was isolated more frequently from goats (21.7% than from other animal groups sampled and isolated more frequently from well water (38.2% than chlorinated supplies (23.0%. A. hydrophilia was the most dominant species isolated from different kinds of samples (13.4%. Whereas A. sobria and A. caviae were isolated in much lower rates 4.7 and 2.1% respectively. There was significant association between the isolation of Aeromonas species from all animal faeces and its presence in drinking water. All isolated strains were examined for the characteristics that are reputed to have roles in pathogenicity. The data reported in this study indicates that the distributions of virulence factors, that regulate the pathogenicity of Aeromonads, are different in clinical and enviromental samples. Aeromonas isolates exhibited multi-drug resistanc amoxicillin, carbenicillin and ampicillin. The most potent antibiotics against Aeromonas species isolated in this study were ceftriaxone, ceftazidime, cefotaxime, cefepime. Essential oils have been tested for in vitro and in vivo antimicrobial activity. Clove, Olive and Peppermint oil exhibited a wide spectrum of antimicrobial activity against all strains used in this study, showed a zone of inhibition ranging from 10.00±0.8 to 14.82±0.41 mm in diameter. Minimum Inhibitory Concentration (MIC for selected oils ranged from 12.8 to 25.6 mg mL-1. Treatment of mice with essential oil for 15 days led to enhance antibody levels in all treated groups and significant clearance of A. hydrophilia from animals. The treated animals had

  3. Aeromonas spp. e Plesiomonas shigelloides isoladas a partir de mexilhões (Perna perna in natura e pré-cozidos no Rio de Janeiro, RJ Aeromonas spp. and Plesiomonas shigelloides isolated from in natura and precooked mussels (Perna perna in Rio de Janeiro, RJ, Brazil

    Directory of Open Access Journals (Sweden)

    Christiane Soares Pereira

    2004-12-01

    Full Text Available O ecossistema aquático é o habitat de mexilhões (Perna perna, animais filtradores que refletem a qualidade ambiental através de análise microbiológica de sua carne. No presente trabalho avaliou-se a presença de patógenos emergentes (Aeromonas hydrophila e Plesiomonas shigelloides, em mexilhões in natura e pré-cozidos coletados por pescadores da Estação Experimental de Cultivo de Mexilhões situada em Jurujuba, Niterói, Rio de Janeiro. Foram analisadas 86 amostras de mexilhões (43 in natura e 43 pré-cozidos as quais foram submetidas a enriquecimento em Água Peptonada Alcalina (APA acrescida de 1 e 3% de Cloreto de Sódio (NaCl e em solução Salina de Butterfield, incubadas a 37ºC por 24 horas. Em seguida, foram semeadas em Ágar Seletivo para Pseudomonas-Aeromonas (GSP, Ágar Tiossulfato Citrato Bile Sacarose (TCBS e Ágar Inositol Bile Verde Brilhante (IBB. A análise geral dos resultados permitiu a identificação de Areomonas spp e Plesiomonas shigelloides em 86% das amostras de mexilhões in natura e pré-cozidas avaliadas. A posterior caracterização bioquímica permitiu a identificação das espécies Aeromonas media (37,10%, A. hydrophila (15,50%, A. caviae (14,80%, A. veronii biogrupo veronii (11,60%, Aeromonas sp. (7,36%, A. sobria (4,20%, A. trota (4,20%, A. schubertii (1,31%, A. jandaei (1,31%, A. veronii biogrupo sobria (0,52% e Plesiomonas shigelloides (2,10%. A relevância epidemiológica desses microrganismos em casos de gastrenterite humana, após consumo de mexilhões crus ou parcialmente cozidos, revela a importância de alertar as autoridades de Saúde Pública no Brasil, sobre a presença desses patógenos na cadeia alimentar e seus riscos para a saúde humana.The aquatic ecosystem is the habitat of mussels, filtrating animals that reflect the ambient quality through microbiological analysis. In the present investigation, we evaluated the presence of emergent pathogens (Aeromonas hydrophila and Plesiomonas

  4. Experimental induction of motile Aeromonas septicemia in channel catfish by waterborne challenge with virulent Aeromonas hydrophila

    Science.gov (United States)

    Motile Aeromonas septicemia (MAS), caused by virulent clonal isolates of Aeromonas hydrophila (vAh), is emerging as a major disease in catfish aquaculture in the Southeastern United States. Predisposing conditions leading to vAh infection in catfish were however largely unknown. The objective of thi...

  5. Infection and disease progress of motile Aeromonas septicemia caused by virulent Aeromonas hydrophila in channel catfish

    Science.gov (United States)

    Motile Aeromonas septicemia (MAS), caused by virulent clonal isolates of Aeromonas hydrophila (vAh), is emerging as a major disease in channel catfish (Ictalurus punctatus) aquaculture in the Southeastern United States. Predisposing conditions leading to vAh infection in catfish were however largely...

  6. Aeromonas flagella and colonisation mechanisms.

    Science.gov (United States)

    Lowry, Rebecca; Balboa, Sabela; Parker, Jennifer L; Shaw, Jonathan G

    2014-01-01

    Aeromonas species are inhabitants of aquatic environments and are able to cause disease in humans and fish among other animals. In aquaculture, they are responsible for the economically important diseases of furunculosis and motile Aeromonas septicaemia (MAS). Whereas gastroenteritis and wound infections are the major human diseases associated with the genus. As they inhabit and survive in diverse environments, aeromonads possess a wide range of colonisation factors. The motile species are able to swim in liquid environments through the action of a single polar flagellum, the flagellin subunits of which are glycosylated; although essential for function the biological role of glycan addition is yet to be determined. Approximately 60% of aeromonads possess a second lateral flagella system that is expressed in viscous environments for swarming over surfaces; both flagellar systems have been shown to be important in the initial colonisation of surfaces. Subsequently, other non-flagellar colonisation factors are employed; these can be both filamentous and non-filamentous. The aeromonads possess a number of fimbrial systems with the bundle-forming MSHA type IV pilus system, having a major role in human cell adherence. Furthermore, a series of outer-membrane proteins have also been implicated in the aeromonad adhesion process. A number of strains are also capable of cell invasion and that maybe linked with the more invasive diseases of bacteraemia or wound infections. These strains employ cell surface factors that allow the colonisation of these niches that protect them from the host's immune system such as S-layers, capsules or particular lipopolysaccharides.

  7. Interlaboratorium vergelijking van het onderzoek naar Aeromonas spp. in drinkwater

    NARCIS (Netherlands)

    Havelaar AH; During M; Versteegh JFM

    1986-01-01

    Door middel van onderzoek van zes kunstmatig besmette gesimuleerde monsters drinkwater ( 4 Aeromonas spp., 1 Klebsiella oxytoca, 1 Pseudomonas aeruginosa) werd een vergelijking gemaakt van de resultaten van Aeromonas onderzoek in 14 laboratoria. De tellingen in de deelnemende laboratoria vertoonde

  8. Epidemiology of Chlamydophila caviae-like Chlamydia Isolated from Urethra and Uterine Cervix

    Directory of Open Access Journals (Sweden)

    Kariyama,Reiko

    2010-02-01

    Full Text Available In 2000, chlamydial strains OK133 and OK135 were isolated from 2 female patients with cervicitis. These strains were unresponsive to commercially available PCR and LCR test kits for the diagnosis of Chlamydia trachomatis infection, and their phenotypic characteristics were very similar. The OK135 nucleotide sequence in MOMP-VD2 gene closely resembled that of Chlamydophila caviae GPIC. A similar strain was isolated in 2003 from a male patient OKM2 with urethritis, from which the strain SC10-6 was cloned by the plaque purification method. The nucleotide sequence of the entire MOMP gene of SC10-6 was exactly the same as that of OK135. Thus, the strains OK135 and SC10-6, together with OK133, have been called C. caviae-like Chlamydia. We designed primers for nested PCR assay, the product of which showed a single-band 311-bp fragment, to detect C. caviae-like Chlamydia. Of swab specimens obtained from 202 patients from 2003 to 2006 (119 male and 83 female patients, 18 specimens (8.9% from 14 male and 4 female patients were positive, suggesting that C. caviae-like Chlamydia infection is rather common. Thus far, it has not been determined whether C. caviae-like Chlamydia is pathogenic for humans.

  9. Epidemiology of Chlamydophila caviae-like Chlamydia isolated from urethra and uterine cervix.

    Science.gov (United States)

    Murao, Wataru; Wada, Koichiro; Matsumoto, Akira; Fujiwara, Michihisa; Fukushi, Hideto; Kishimoto, Toshio; Monden, Koichi; Kariyama, Reiko; Kumon, Hiromi

    2010-02-01

    In 2000, chlamydial strains OK133 and OK135 were isolated from 2 female patients with cervicitis. These strains were unresponsive to commercially available PCR and LCR test kits for the diagnosis of Chlamydia trachomatis infection, and their phenotypic characteristics were very similar. The OK135 nucleotide sequence in MOMP-VD2 gene closely resembled that of Chlamydophila caviae GPIC. A similar strain was isolated in 2003 from a male patient OKM2 with urethritis, from which the strain SC10-6 was cloned by the plaque purification method. The nucleotide sequence of the entire MOMP gene of SC10-6 was exactly the same as that of OK135. Thus, the strains OK135 and SC10-6, together with OK133, have been called C. caviae-like Chlamydia. We designed primers for nested PCR assay, the product of which showed a single-band 311-bp fragment, to detect C. caviae-like Chlamydia. Of swab specimens obtained from 202 patients from 2003 to 2006 (119 male and 83 female patients), 18 specimens (8.9%) from 14 male and 4 female patients were positive, suggesting that C. caviae-like Chlamydia infection is rather common. Thus far, it has not been determined whether C. caviae-like Chlamydia is pathogenic for humans.

  10. Characterization of mercury-resistant clinical Aeromonas species

    Directory of Open Access Journals (Sweden)

    Abigail Pérez-Valdespino

    2013-12-01

    Full Text Available Mercury-resistant Aeromonas strains isolated from diarrhea were studied. Resistance occurs via mercuric ion reduction but merA and merR genes were only detected in some strains using PCR and dot hybridization. Results indicate a high variability in mer operons in Aeromonas. To our knowledge, this is the first report of mercury-resistant clinical Aeromonas strains.

  11. Characterization of mercury-resistant clinical Aeromonas species.

    Science.gov (United States)

    Pérez-Valdespino, Abigail; Celestino-Mancera, Martin; Villegas-Rodríguez, Viridiana Lorena; Curiel-Quesada, Everardo

    2013-12-01

    Mercury-resistant Aeromonas strains isolated from diarrhea were studied. Resistance occurs via mercuric ion reduction but merA and merR genes were only detected in some strains using PCR and dot hybridization. Results indicate a high variability in mer operons in Aeromonas. To our knowledge, this is the first report of mercury-resistant clinical Aeromonas strains.

  12. High prevalence of blaCTX-M group genes in Aeromonas dhakensis isolated from aquaculture fish species in South Korea.

    Science.gov (United States)

    Yi, Seung-Won; Chung, Tae-Ho; Joh, Seong-Joon; Park, Chul; Park, Byoung-Yong; Shin, Gee-Wook

    2014-12-01

    The prevalence of resistant genes against β-lactams in 119 Aeromonas strains was determined. A large number (99.2%) of the present fish strains were resistant to one or more β- lactams including ceftiofur, amoxicillin-clavulanic acid, ampicillin, piperacillin and cefpodoxime. Among antibiotic resistance phenotypes, the simultaneous resistance to all β-lactams occurred in 25.2% (n=30) of all strains, which consisted of 18 strains of A. dhakensis, 8 strains of A. caviae, 2 strains of A. hydrophila and only one strain of A. veronii. For exploring genetic background of the antibiotic resistances, multiple PCR assays were subjected to detect β-lactamase-encoding genes, bla(TEM), bla(OXA-B) and bla(CTX-M). In the results, the bla(TEM-1) gene was harbored in all strains, whereas only 3 strains harbored bla(OXA) gene. In the case of bla(CTX-M) gene, the gene was detected in 21.0% (25 out of 119) of all strains, which countered with 80% (20 out of 25) of A. dhakensis, 8% (2 out of 25) of A. caviae and 12% (3 out of 25) of A. hydrophila. In addition, most of the bla(CTX-M) positive strains showed simultaneous resistance to all β-lactams (18 out of 30 strains). In sequence analysis for bla(CTX-M) genes detected, they were CTX-M group 1-encoding genes including bla(CTX-M-33) from 3 eel strains of A. dhakensis. Therefore, A. dhakensis obtained from cultured fish could represent a reservoir for spreading genes encoding CTX-M group 1 enzymes and hence should be carefully monitored, especially for its potential risk to public health.

  13. Prolactin family of the guinea pig, Cavia porcellus.

    Science.gov (United States)

    Alam, S M Khorshed; Konno, Toshihiro; Rumi, M A Karim; Dong, Yafeng; Weiner, Carl P; Soares, Michael J

    2010-08-01

    Prolactin (PRL) is a multifunctional hormone with prominent roles in regulating growth and reproduction. The guinea pig (Cavia porcellus) has been extensively used in endocrine and reproduction research. Thus far, the PRL cDNA and protein have not been isolated from the guinea pig. In the present study, we used information derived from the public guinea pig genome database as a tool for identifying guinea pig PRL and PRL-related proteins. Guinea pig PRL exhibits prominent nucleotide and amino acid sequence differences when compared with PRLs of other eutherian mammals. In contrast, guinea pig GH is highly conserved. Expression of PRL and GH in the guinea pig is prominent in the anterior pituitary, similar to known expression patterns of PRL and GH for other species. Two additional guinea pig cDNAs were identified and termed PRL-related proteins (PRLRP1, PRLRP2). They exhibited a more distant relationship to PRL and their expression was restricted to the placenta. Recombinant guinea pig PRL protein was generated and shown to be biologically active in the PRL-responsive Nb2 lymphoma cell bioassay. In contrast, recombinant guinea pig PRLRP1 protein did not exhibit PRL-like bioactivity. In summary, we have developed a new set of research tools for investigating the biology of the PRL family in an important animal model, the guinea pig.

  14. Development of duplex-PCR for identification of Aeromonas species

    Directory of Open Access Journals (Sweden)

    Carina Lucena Mendes-Marques

    2013-10-01

    Full Text Available Introduction The number of reports of intestinal infections caused by Aeromonas spp. has increased significantly in recent years. In most clinical laboratories, identification of these bacteria is carried out by general phenotypic tests that sometimes do not accurately differentiate Aeromonas and Vibrio. Methods A duplex-polymerase chain reaction (PCR was developed directed to 2 targets identifying Aeromonas spp. pathogenic to humans. Results The duplex-PCR results were reproducible and specific for Aeromonas spp. pathogenic to humans. Conclusions This method will allow differentiation between Vibrio and Aeromonas spp. in patients with in cholera-like symptoms and can also be used in water quality monitoring.

  15. Development of duplex-PCR for identification of Aeromonas species

    Directory of Open Access Journals (Sweden)

    Carina Lucena Mendes-Marques

    2013-05-01

    Full Text Available Introduction The number of reports of intestinal infections caused by Aeromonas spp. has increased significantly in recent years. In most clinical laboratories, identification of these bacteria is carried out by general phenotypic tests that sometimes do not accurately differentiate Aeromonas and Vibrio. Methods A duplex-polymerase chain reaction (PCR was developed directed to 2 targets identifying Aeromonas spp. pathogenic to humans. Results The duplex-PCR results were reproducible and specific for Aeromonas spp. pathogenic to humans. Conclusions This method will allow differentiation between Vibrio and Aeromonas spp. in patients with in cholera-like symptoms and can also be used in water quality monitoring.

  16. Aeromonas phages encode tRNAs for their overused codons.

    Science.gov (United States)

    Prabhakaran, Ramanandan; Chithambaram, Shivapriya; Xia, Xuhua

    2014-01-01

    The GC-rich bacterial species, Aeromonas salmonicida, is parasitised by both GC-rich phages (Aeromonas phages - phiAS7 and vB_AsaM-56) and GC-poor phages (Aeromonas phages - 25, 31, 44RR2.8t, 65, Aes508, phiAS4 and phiAS5). Both the GC-rich Aeromonas phage phiAS7 and Aeromonas phage vB_AsaM-56 have nearly identical codon usage bias as their host. While all the remaining seven GC-poor Aeromonas phages differ dramatically in codon usage from their GC-rich host. Here, we investigated whether tRNA encoded in the genome of Aeromonas phages facilitate the translation of phage proteins. We found that tRNAs encoded in the phage genome correspond to synonymous codons overused in the phage genes but not in the host genes.

  17. Herida antebraquial infectada por Aeromonas hydrophila

    OpenAIRE

    1995-01-01

    Se presenta un caso de celulitis por Aeromonas hydrophila, en un paciente de 23 años, tras una herida antebraquial producida en el medio acuático. La celulitis se resolvió mediante desbridamiento agresivo y quimioterapia. En los controles posteriores no se han apreciado déficits funcionales tras la infección. A case of Aeromonas hidrophila cellulitis developped in a 23-year-old patient, after aquatic environment forearm wound is presented. His cellulitis was effectively resolve...

  18. Paraspidodera uncinata (Nematoda, Lauroiinae as parasite of Cavia magna and Cavia aperea (Rodentia, Caviidae in southern Brazil

    Directory of Open Access Journals (Sweden)

    Ana Gabriela da Silva Rocha

    2015-05-01

    Full Text Available Out of the 5 species of Guinea pig registered for Brazil, 4 (Cavia fulgida, C. porcellus, C. aperea, and C. magna have already been reported as hosts of Paraspidodera uncinata. The rodent species C. magna and C. aperea are small-sized mammals, with terrestrial habits, which occur from southern Brazil to eastern Uruguay. Guinea pig specimens donated for research were necropsied for the analysis of endoparasites, and the nematodes found were identified as P. uncinata. The microhabitats of this species were the small and large intestines in C. magna and the large intestine in C. aperea. The prevalence of P. uncinata in the hosts was 60% in C. magna (n = 5 and 14% in C. aperea (n = 7. The record of P. uncinata in different Guinea pig species allows inferring that the foraging mode is similar in the different locations where they occur in southern Brazil, because, perhaps, the infection of hosts has occurred through the ingestion of vegetables contaminated with P. uncinata eggs. This study contributes to knowledge on the helminth fauna of C. magna and C. aperea in southern Brazil.

  19. Aeromonas spp. simultaneously harbouring bla(CTX-M-15), bla(SHV-12), bla(PER-1) and bla(FOX-2), in wild-growing Mediterranean mussel (Mytilus galloprovincialis) from Adriatic Sea, Croatia.

    Science.gov (United States)

    Maravić, Ana; Skočibušić, Mirjana; Samanić, Ivica; Fredotović, Zeljana; Cvjetan, Svjetlana; Jutronić, Marinka; Puizina, Jasna

    2013-09-02

    Aeromonas species are becoming renowned as emerging pathogens by increasingly giving rise to a wide spectrum of food and waterborne infections in humans. Another worrisome feature of aeromonads is the growing frequency of antibiotic resistance as a consequence of their prominent diversity in terms of resistance determinants. This study aimed at determining the antimicrobial resistance pattern, prevalence and characterization of acquired β-lactamases, including extended-spectrum-β-lactamases (ESBLs) and AmpC cephalosporinases, as well as the presence of class 1 and 2 integrons, in Aeromonas isolates from wild-growing Mediterranean mussel (Mytilus galloprovincialis) of the eastern coast of Adriatic Sea, Croatia. Isolates were tested for susceptibility to 16 antibiotics and β-lactam/β-lactamase inhibitor combinations. Cephalosporin-resistant isolates were further screened by PCR for genes encoding AmpC (bla(FOX), bla(CMY), bla(MOX), bla(LAT), bla(BIL), bla(DHA), bla(ACC), bla(MIR), bla(ACT)), ESBLs (bla(TEM), bla(SHV), bla(CTX-M), bla(PER), bla(VEB), bla(GES/IBC), bla(OXA)) and integrases (intI1, intI2, intI3). Location of bla genes was characterized by plasmid DNA fingerprinting and Southern blot hybridization. Plasmids carrying ESBL genes were investigated for transferability by conjugation and PCR-based replicon typed. Out of 147 Aeromonas isolates recovered, 30 (20%) demonstrated multiple resistance profile, with co-resistance most frequently detected against penicillins, piperacillin/sulbactam and tetracycline. ESBL-encoding genes were detected in 21 (13 Aeromonas caviae and 8 Aeromonas hydrophila) isolates, with bla(CTX-M-15) gene identified in 19 and bla(SHV-12) in 12 isolates. Among them, 10 isolates simultaneously harboured bla(CTX-M-15) and bla(SHV-12), while 3 isolates additionally carried an AmpC β-lactamase bla(FOX-2) gene. bla(PER-1) gene was identified in a single isolate also harbouring the bla(CTX-M-15) gene. While bla(SHV-12) was chromosomally

  20. Identification of Aeromonas hydrophila Genes Preferentially Expressed after Phagocytosis by Tetrahymena and Involvement of Methionine Sulfoxide Reductases

    Science.gov (United States)

    Pang, Maoda; Lin, Xiaoqin; Liu, Jin; Guo, Changming; Gao, Shanshan; Du, Hechao; Lu, Chengping; Liu, Yongjie

    2016-01-01

    Free-living protozoa affect the survival and virulence evolution of pathogens in the environment. In this study, we explored the fate of Aeromonas hydrophila when co-cultured with the bacteriovorous ciliate Tetrahymena thermophila and investigated bacterial gene expression associated with the co-culture. Virulent A. hydrophila strains were found to have ability to evade digestion in the vacuoles of this protozoan. In A. hydrophila, a total of 116 genes were identified as up-regulated following co-culture with T. thermophila by selective capture of transcribed sequences (SCOTS) and comparative dot-blot analysis. A large proportion of these genes (42/116) play a role in metabolism, and some of the genes have previously been characterized as required for bacterial survival and replication within macrophages. Then, we inactivated the genes encoding methionine sulfoxide reductases, msrA, and msrB, in A. hydrophila. Compared to the wild-type, the mutants ΔmsrA and ΔmsrAB displayed significantly reduced resistance to predation by T. thermophila, and 50% lethal dose (LD50) determinations in zebrafish demonstrated that both mutants were highly attenuated. This study forms a solid foundation for the study of mechanisms and implications of bacterial defenses. PMID:28083518

  1. Learning and personality types are related in cavies (Cavia aperea).

    Science.gov (United States)

    Guenther, Anja; Brust, Vera; Dersen, Mona; Trillmich, Fritz

    2014-02-01

    The evolution and maintenance of consistent individual differences, so called animal personalities, have attracted much research interest over the past decades. Variation along common personality traits, such as boldness or exploration, is often associated with risk-reward trade-offs. Individuals that are bolder and hence take more risks may be more successful in acquiring resources over the short term. Cautious individuals taking fewer risks may, on the other hand, live longer, but may also gather fewer resources over the short term. According to recent theory, individual differences in personality may be functionally related to individual differences in cognitive performance (i.e., the way in which individuals acquire or use information). Individual differences in the acquisition speed of cognitively challenging tasks are often associated with a speed-accuracy trade-off. Accuracy can be improved by investing more time in the decision-making process or, conversely, decisions can be made more quickly at the cost of making more mistakes. Hence, the speed-accuracy trade-off often involves a risk-reward trade-off. We tested whether 3 personality traits, boldness, activity, and aggressiveness, are correlated with individual learning, associative learning speed, and behavioral flexibility as assessed by reversal learning in wild cavies (Cavia aperea). We found strong positive relationships between all personality traits and learning speed, whereas flexibility was negatively associated with aggressiveness. Our results support the hypothesis that performance reflects individual differences in personality in a predictable way. PsycINFO Database Record (c) 2014 APA, all rights reserved.

  2. Quorum Sensing Regulation in Aeromonas hydrophila

    DEFF Research Database (Denmark)

    Garde, Christian; Bjarnsholt, Thomas; Givskov, Michael;

    2010-01-01

    We present detailed results on the C4-HSL-mediated quorum sensing (QS) regulatory system of the opportunistic Gram-negative bacterium Aeromonas hydrophila. This bacterium contains a particularly simple QS system that allows for a detailed modeling of kinetics. In a model system (i...

  3. Multilocus phylogenetic analysis of the genus Aeromonas.

    Science.gov (United States)

    Martinez-Murcia, Antonio J; Monera, Arturo; Saavedra, M Jose; Oncina, Remedios; Lopez-Alvarez, Monserrate; Lara, Erica; Figueras, M Jose

    2011-05-01

    A broad multilocus phylogenetic analysis (MLPA) of the representative diversity of a genus offers the opportunity to incorporate concatenated inter-species phylogenies into bacterial systematics. Recent analyses based on single housekeeping genes have provided coherent phylogenies of Aeromonas. However, to date, a multi-gene phylogenetic analysis has never been tackled. In the present study, the intra- and inter-species phylogenetic relationships of 115 strains representing all Aeromonas species described to date were investigated by MLPA. The study included the independent analysis of seven single gene fragments (gyrB, rpoD, recA, dnaJ, gyrA, dnaX, and atpD), and the tree resulting from the concatenated 4705 bp sequence. The phylogenies obtained were consistent with each other, and clustering agreed with the Aeromonas taxonomy recognized to date. The highest clustering robustness was found for the concatenated tree (i.e. all Aeromonas species split into 100% bootstrap clusters). Both possible chronometric distortions and poor resolution encountered when using single-gene analysis were buffered in the concatenated MLPA tree. However, reliable phylogenetic species delineation required an MLPA including several "bona fide" strains representing all described species.

  4. Aeromonas spp.: ubiquitous or specialized bugs?

    Science.gov (United States)

    Martino, Maria Elena; Fasolato, Luca; Montemurro, Filomena; Novelli, Enrico; Cardazzo, Barbara

    2014-04-01

    The genus Aeromonas comprises ubiquitous bacteria that are known to play several roles in the environment. These bacteria were first described as fish pathogens, but their presence was documented in other reservoirs, such as animals and humans. Today, these bacteria are described as emerging pathogens, but their effective role in human pathogenicity is still controversial. In addition, their taxonomy is heavily debated, as species distinction is often difficult to achieve. To study the interspecies relationships and to investigate their connection with the environment, a multilocus sequence typing scheme previously developed for Aeromonas spp. was applied to 258 strains, and the genetic data were analysed by population software. Sampling was a fundamental step, including several of the main sources of Aeromonas: fish, food products and human cases of disease. The objective was to characterize the isolates and to find potential associations among them according to the following: species, sharing of virulence factors, source and adaptation to a specific habitat. The strains were characterized and demonstrated exceptionally high nucleotide variability in the Aeromonas genus. Among the sampled sources, different species distributions were found, highlighting the occurrence of adaptation processes towards specific habitats.

  5. Aeromonas chitinase degrades chironomid egg masses.

    Science.gov (United States)

    Laviad, Sivan; Golan, Amnon; Shaked, Tamar; Vaizel-Ohayon, Dalit; Halpern, Malka; Pick, Elah

    2016-02-01

    Chironomids are freshwater insects that undergo a complete metamorphosis of four life stages. Chironomid egg masses can be degraded by Vibrio cholerae and some Aeromonas species. Egg mass degradation by V. cholerae requires haemagglutinin protease activity. Our aim was to identify the egg mass degrading (EMD) factor secreted by Aeromonas dhkanesis 3K1C15. Following the hypothesis that the EMD factor of A. dhkanesis is also a protease, secreted proteases were screened, but none of them proved to have the same properties as the EMD factor. Using conventional protein purification methods, we found that the active fraction included chitinases. We further confirmed chitin as a building block of the egg masses. Interestingly, by supplementing bacterial growth media with chitin, we observed unexpected EMD factor activity in Aeromonas isolates that initially were not able to degrade egg masses. Accordingly, we concluded that although strain 3K1C15 secretes chitinases constitutively, most Aeromonas strains secrete chitinases inductively. Induction of chitinases in nature presumably occurs when bacteria are attached to the egg mass habitat, in which chitin is abundant. Considering that chitinases are highly conserved across bacteria phyla, we assume that the role of this enzyme in the bacteria-insect interplay could be wider than is currently thought.

  6. Chlamydia caviae infection alters abundance but not composition of the guinea pig vaginal microbiota.

    Science.gov (United States)

    Neuendorf, Elizabeth; Gajer, Pawel; Bowlin, Anne K; Marques, Patricia X; Ma, Bing; Yang, Hongqiu; Fu, Li; Humphrys, Michael S; Forney, Larry J; Myers, Garry S A; Bavoil, Patrik M; Rank, Roger G; Ravel, Jacques

    2015-06-01

    In humans, the vaginal microbiota is thought to be the first line of defense again pathogens including Chlamydia trachomatis. The guinea pig has been extensively used as a model to study chlamydial infection because it shares anatomical and physiological similarities with humans, such as a squamous vaginal epithelium as well as some of the long-term outcomes caused by chlamydial infection. In this study, we aimed to evaluate the guinea pig-C. caviae model of genital infection as a surrogate for studying the role of the vaginal microbiota in the early steps of C. trachomatis infection in humans. We used culture-independent molecular methods to characterize the relative and absolute abundance of bacterial phylotypes in the guinea pig vaginal microbiota in animals non-infected, mock-infected or infected by C. caviae. We showed that the guinea pig and human vaginal microbiotas are of different bacterial composition and abundance. Chlamydia caviae infection had a profound effect on the absolute abundance of bacterial phylotypes but not on the composition of the guinea pig vaginal microbiota. Our findings compromise the validity of the guinea pig-C. caviae model to study the role of the vaginal microbiota during the early steps of sexually transmitted infection.

  7. Do young guinea pigs (Cavia porcellus) develop an attachment to inanimate objects?

    NARCIS (Netherlands)

    Janzen, MID; Timmermans, PJA; Kruijt, JP; Vossen, JMH

    1999-01-01

    Filial imprinting has been studied extensively in precocial birds. In these studies, inanimate objects were used as imprinting objects. Although attachment to the parents is common in mammals, experiments with inanimate objects are rare and mostly restricted to guinea pigs (Cavia porcellus). The res

  8. MONITORING FOR AEROMONAS SPECIES AFTER TREATMENT WITH COMMON DRINKING WATER DISINFECTANTS

    Science.gov (United States)

    The sensitivity of Aeromonas spp. To free chlorine, chloramine and ultraviolet (UV) disinfection was determined. Aeromonas hydrophila is a contaminant listed on the USEPA's 1998 Contaminant Candidate List (CCL). Experiments using free chlorine indicated that the Aeromonas spp. ...

  9. MONITORING FOR AEROMONAS SPECIES AFTER TREATMENT WITH COMMON DRINKING WATER DISINFECTANTS

    Science.gov (United States)

    The sensitivity of Aeromonas spp. To free chlorine, chloramine and ultraviolet (UV) disinfection was determined. Aeromonas hydrophila is a contaminant listed on the USEPA's 1998 Contaminant Candidate List (CCL). Experiments using free chlorine indicated that the Aeromonas spp. ...

  10. Determinación de factores de virulencia en cepas de Aeromonas spp., aisladas a partir de pescado

    Directory of Open Access Journals (Sweden)

    William Suárez Q.

    2012-04-01

    Full Text Available Objetivo. Investigar la incidencia de cinco marcadores fenotípicos de virulencia en cepas de Aeromonas aisladas a partir de muestras de pescado expendido en Pamplona, Colombia. Materiales y métodos. Se utilizaron 47 cepas identificadas previamente. Se evaluaron: actividad hemolítica en agar sangre, suplementado con 5% de eritrocitos de cordero y agar sangre suplementado con 5% eritrocitos de sangre humana; actividad proteolítica en agar Mueller-Hinton suplementado al 10% (p/v con leche descremada, actividad lipolítica en agar tributirina; actividad desoxirribonucleasas en agar DNAsa. Resultados. Se encontró que las cepas de A. hydrophila, A. veronii GH 8, A. jandaei, A. veronii GH 10 y A. eucrenophila, demostraron capacidad hemolítica, proteolítica, lipolítica y nucleasa. Todas las cepas de A. popoffii fueron β-hemolíticas en agar sangre humana, proteolíticas y con actividad DNAsa. Las cepas de A. caviae, coincidieron en ser hemolíticas y lipolitícas, mientras que la cepa de A. schubertii, manifestó la presencia de actividad hemolítica y DNAsa. Conclusiones. La frecuencia de los factores de virulencia en las cepas estudiadas fue: el 87% demostraron producción de nucleasas; el 83% fueron β-hemolíticas sobre eritrocitos humanos; el 68% expresaron producción de lipasas, el 63% fueron proteolíticas y el 53% resultaron ser hemolíticas sobre eritrocitos de cordero, indicando estos datos el posible potencial patógeno de las cepas. Estos resultados mostraron que el pescado comercializado en Pamplona, puede ser una fuente importante de especies de Aeromonas que expresan factores asociados a la virulencia para el hombre.

  11. Re-identification of Aeromonas isolates from rainbow trout and incidence of class 1 integron and β-lactamase genes.

    Science.gov (United States)

    Vega-Sánchez, Vicente; Latif-Eugenín, Fadua; Soriano-Vargas, Edgardo; Beaz-Hidalgo, Roxana; Figueras, María José; Aguilera-Arreola, Ma Guadalupe; Castro-Escarpulli, Graciela

    2014-08-27

    Forty-eight Aeromonas isolates from rainbow trout previously identified by the 16S rDNA-RFLP technique were re-identified using 2 housekeeping genes (gyrB and rpoD). After sequencing the prevalences of the species were A. veronii (29.2%), A. bestiarum (20.8%), A. hydrophila (16.7%), A. sobria (10.4%), A. media (8.3%), A. popoffii (6.2%), A. allosaccharophila (2.1%), A. caviae (2.1%), A. salmonicida (2.1%) and one isolate (2.1%) belongs to a candidate new species "Aeromonas lusitana". Coincident identification results to the 16S rDNA-RFLP technique were only obtained for 68.8% of the isolates. PCR amplification of the enterobacterial repetitive intergenic consensus (ERIC-PCR) indicated that the 48 isolates belonged to 33 different ERIC genotypes. Several genotypes were isolated from different farms and organs in the same fish, indicating a systemic dissemination of the bacteria. The presence of genes (blaIMP, blaCphA/IMIS, blaTEM, blaSHV and intI1) that encode extended-spectrum beta-lactamases (ESBLs), metallo-beta-lactamases (MBLs) and class 1 integrons were studied by PCR. Only 39.6% (19/48) of the strains showed the presence of one or more resistance genes. The gene blaCphA/IMIS was detected in 29.2% of the isolates, followed by the intI1 (6.2%) and blaSHV (4.2%) genes. The variable region of class 1 integrons of the 3 positive isolates was sequenced revealing the presence of the gene cassette aadA1 (aminoglycoside transferase) that plays a role in streptomycin/spectinomycin resistance.

  12. Sanitary evaluation of domestic water supply facilities with storage tanks and detection of Aeromonas, enteric and related bacteria in domestic water facilities in Okinawa Prefecture of Japan.

    Science.gov (United States)

    Miyagi, Kazufumi; Sano, Kouichi; Hirai, Itaru

    2017-08-01

    To provide for temporary restrictions of the public water supply system, storage tanks are commonly installed in the domestic water systems of houses and apartment buildings in Okinawa Prefecture of Japan. To learn more about the sanitary condition and management of these water supply facilities with storage tanks (hereafter called "storage tank water systems") and the extent of bacterial contamination of water from these facilities, we investigated their usage and the existence of Aeromonas, enteric and related bacteria. Verbal interviews concerning the use and management of the storage tank water systems were carried out in each randomly sampled household. A total of 54 water samples were collected for bacteriological and physicochemical examinations. Conventional methods were used for total viable count, fecal coliforms, identification of bacteria such as Aeromonas, Enterobacteriaceae and non-fermentative Gram-negative rods (NF-GNR), and measurement of residual chlorine. On Aeromonas species, tests for putative virulence factor and an identification using 16S rRNA and rpoB genes were also performed. Water from the water storage systems was reported to be consumed directly without boiling in 22 of the 54 houses (40.7%). 31 of the sampled houses had installed water storage tanks of more than 1 cubic meter (m(3)) per inhabitant, and in 21 of the sampled houses, the tank had never been cleaned. In all samples, the total viable count and fecal coliforms did not exceed quality levels prescribed by Japanese waterworks law. Although the quantity of bacteria detected was not high, 23 NF-GNR, 14 Enterobacteriaceae and 5 Aeromonas were isolated in 42.6%, 7.4% and 3.7% of samples respectively. One isolated A. hydrophila and four A. caviae possessed various putative virulence factors, especially A. hydrophila which had diverse putative pathogenic genes such as aer, hlyA, act, alt, ast, ser, and dam. Many bacteria were isolated when the concentration of residual chlorine

  13. Microalgae harvesting via co-culture with filamentous fungus

    Science.gov (United States)

    Gultom, Sarman Oktovianus

    Microalgae harvesting is a labor- and energy-intensive process. For instance, classical harvesting technologies such as chemical addition and mechanical separation are economically prohibiting for biofuel production. Newer approaches to harvest microalgae have been developed in order to decrease costs. Among these new methods, fungal co-pelletization seems to be a promising technology. By co-culturing filamentous fungi with microalgae, it is possible to form pellets, which can easily be separated. In this study, different parameters for the cultivation of filamentous fungus (Aspergillus niger) and microalgae (Chlorella vulgaris) to efficiently form cell pellets were evaluated under heterotrophic and phototrophic conditions, including organic carbon source (glucose, glycerol and sodium acetate) concentration, pH, initial concentration of fungal spores, initial concentration of microalgal cells, concentration of ionic strength (Calcium and Magnesium) and concentration of salinity (NaCl). In addition, zeta-potential measurements were carried out in order to get a better understanding of the mechanism of attraction. It was found that 2 g/L of glucose, a fungus to microalgae ratio of 1:300, and uncontrolled pH (around 7) are the best culturing conditions for co-pelletization. Under these conditions, it was possible to achieve a high harvesting performance (>90%). In addition, it was observed that most pellets formed in the co-culture were spherical with an average diameter of 3.5 mm and in concentrations of about 5 pellets per mL of culture media. Under phototrophic conditions, co-pelletization required the addition of glucose as organic carbon source to sustain the growth of fungi and to allow the harvesting of microalgae. Zeta-potential measurements indicated that (i) both microalgae and fungi have low zeta-potential values regardless of the pH on the bulk (i.e. <-10 mV) (ii) fungi can have a positive electric charge at low pH (ie. pH=3). These values suggest that it

  14. Prevalence and antimicrobial resistance analysis of Aeromonas in Beijing%北京地区气单胞菌的流行状况及耐药性分析

    Institute of Scientific and Technical Information of China (English)

    陈素明; 鲍春梅; 崔恩博; 郭桐生; 张鞠玲; 王欢; 曲芬

    2011-01-01

    Objective To enhance the knowledge of distribution feature and antibiotic resistance in Aeromonas species isolated from infected patients on pastients in order to offer the scientific gist for clinical analysing pathogenetic condition and adopting antibiotics treatment. Methods Bacteria were isolated and identified from samples in infected patients with clinical manifestation. Furthermore the sensitivity of Aeromonas to antimicrobial agents was measured by K-B according to CLSI and the distribution feature and antibiotic resistance of various Aeromonas species were analyzed. Results Aeromonas infected happened in enteron frequently, following up blood and ascites. Among 280 cases of Aeromonas, 87.86% occurred in summer and autumn. Aeromonas sobria was the most common species (40%) and Aeromonas hydrophila was secondary (28.22%). Others were Aeromonas caviae (20.36%), Aeromonas veronni (5.71%) and few species. The resistant rate to antimicrobiai of Aeromonas species isolated from enteron was higher in ampicillin (87.9%), trimethoprim-sulfamethoxazole (61.3%), ceftriaxone (29.4%),cefmetazole (28.6%). The resistant rate to ceftriaxone, cefmetazole, ciprofloxacin of Aeromonas hydrophila and Aeromonas caviae was overtopping that of Aeromonas sobria. The resistance rate to ceftriaxone, levofloxacin, and trimethoprim-sulfamethoxazole of Aeromonas species isolated enteron was higher than that of blood and ascites.Conclusion Aeromonas sobria and Aeromonas hydrophila are majority species and occur in summer and autumn frequently in Beijing. The different resistant rate to some antimicrobial exist in various species and infected site so patients should be treated differently according to the susceptibility test.%目的 了解临床分离气单胞菌感染的分布特点及耐药性状况,为临床评估病情及合理选择抗菌药物提供客观依据.方法 有感染临床表现患者的标本进行培养、分离和鉴定,同时用纸片扩散法测定

  15. Occurrence of bacteriophages infecting Aeromonas, Enterobacter, and Klebsiella in water and association with contamination sources in Thailand.

    Science.gov (United States)

    Wangkahad, Bencharong; Bosup, Suchada; Mongkolsuk, Skorn; Sirikanchana, Kwanrawee

    2015-06-01

    The co-residence of bacteriophages and their bacterial hosts in humans, animals, and environmental sources directed the use of bacteriophages to track the origins of the pathogenic bacteria that can be found in contaminated water. The objective of this study was to enumerate bacteriophages of Aeromonas caviae (AecaKS148), Enterobacter sp. (EnspKS513), and Klebsiella pneumoniae (KlpnKS648) in water and evaluate their association with contamination sources (human vs. animals). Bacterial host strains were isolated from untreated wastewater in Bangkok, Thailand. A double-layer agar technique was used to detect bacteriophages. All three bacteriophages were detected in polluted canal samples, with likely contamination from human wastewater, whereas none was found in non-polluted river samples. AecaKS148 was found to be associated with human fecal sources, while EnspKS513 and KlpnKS648 seemed to be equally prevalent in both human and animal fecal sources. Both bacteriophages were also present in polluted canals that could receive contamination from other fecal sources or the environment. In conclusion, all three bacteriophages were successfully monitored in Bangkok, Thailand. This study provided an example of bacteriophages for potential use as source identifiers of pathogen contamination. The results from this study will assist in controlling sources of pathogen contamination, especially in developing countries.

  16. Occurrence and antibiotic resistance of mesophilic Aeromonas in three riverine freshwaters of Marrakech, Morocco.

    Science.gov (United States)

    Imziln, B

    2001-12-01

    In order to evaluate the impact of pollution and sewage on the occurrence and antibiotic resistance of mesophilic aeromonads in riverine freshwaters of Marrakech, samples were collected from three rivers (Oukaimeden, Ourika, and Tensift) upstream and downstream from the principal bordering villages. During a 2-year study, indicators of pollution increased dramatically in the downstream waters. Bacterial indicators (faecal coliforms and faecal streptococci) correlated with mesophilic aeromonads only in heavily polluted waters. In low and moderately polluted sources, densities of mesophilic aeromonads were independent of water quality indicators and did not correlate statistically with faecal indicators. Average counts of Aeromonas in low and heavily polluted waters were 2.5 x 10(3) and 2.1 x 10(6) colony forming units per 100 ml, respectively. The biochemical identification of 841 isolates indicated a predominance of A. caviae in heavily and moderately polluted water and sediment. A. hydrophila was dominant only in low polluted waters and when the temperature was below 12 degrees C. High densities of A. sobria were found in low, moderately polluted, or cleaned waters and when the water temperature was above 18 degrees C. All selected isolates (total = 841) were tested for antibiotic susceptibility against 21 antibiotics. Antibiotic resistance frequencies recorded were: ampicillin and amoxicillin, 100%; novobiocin, 96%; cefalotin, 81%; colistin, 72%; sulfamethoxazole, 40%; cefamandole, 37%; polymyxin B, 23%; trimethoprim, 17%; erythromycin, 15%; streptomycin, 8%; amoxicillin-clavulanate, 5%. Resistance to cefotaxime, kanamycin, gentamycin, chloramphenicol, tetracycline, oxytetracycline, nalidixic acid, rifampicin, or trimethoprim-sulfameth-oxazole was found to be polluted samples. Since no correlation between mesophilic aeromonads and conventional faecal pollution indicators was observed in low or moderately polluted waters, and since these freshwaters are used for

  17. Resistance of Salmonds Against Aeromonas Salmonicida

    DEFF Research Database (Denmark)

    Holten-Andersen, Lars; Dalsgaard, Inger; Buchmann, Kurt

    2011-01-01

    Furunculosis, caused by Aeromonas salmonicida, continues to be a health problem for the growing salmonid aquaculture. Despite effective vaccination programs regular outbreaks occur at Danish trout farms calling for repeated antibiotic treatment. We hypothesized that a difference in natural...... susceptibility to this disease might exist between Baltic salmon and the widely used rainbow trout. Hence, a cohabitation challenge model was applied to investigate the relative susceptibility to infection with Aeromonas salmonicida in rainbow trout and Baltic salmon. The course of infection was monitored daily...... over a 30-day period post challenge and the results were summarized in mortality curves. A. salmonicida was recovered from mortalities during the entire test period. At day 30 the survival was 6.2 % and 34.0 % for rainbow trout and Baltic salmon, respectively. Significant differences in susceptibility...

  18. Aeromonas Hydrophila: A Re-Emerging Pathogen

    Digital Repository Service at National Institute of Oceanography (India)

    Lakshmanaperumalsamy, P.; Thayumanavan, T.; Subashkumar, R.

    been initiated by their importance as fish pathogens or as a potential pathogen of humans and have, therefore, tended to focus on commercial and sport fisheries and recreational waters. Mesophilic aeromonads are halotolerant and are associated... role of species of A. hydrophila as a cause of human infections. During monsoon season, increased incidence of A. hydrophila was reported (Vivekanandhan et al., 2004). The much higher incidence of Aeromonas sp. in foods purchased at retail outlets...

  19. Aeromonas hydrophila infection in Xenopus laevis.

    Science.gov (United States)

    Hubbard, G B

    1981-06-01

    Aeromonas hydrophila caused severe disease in a group of Xenopus laevis within 3 weeks of receipt. The primary clinical signs were marked pallor of the skin, petechiation, lethargy, anorexia, and edema. The duration of the outbreak was approximately 45 days during which time 21 frogs became sick and 18 died, despite tetracycline therapy. Pale skin, subcutaneous edema and hemorrhages, ascites, and pale livers were seen at necropsy. Aeromonas hydrophila was isolated from skin, and the same organism was isolated in pure culture from skeletal muscle. Tetracycline treatment via stomach tube was effective if given early in the course of the disease. The outbreak was controlled by removing sick frogs, feeding twice per week, changing the water several hours after feeding, and maintaining the frogs where the ambient temperature was 22 degrees C or lower. The pallor of the skin and general malaise were produced experimentally by crowding normal frogs, changing the water infrequently, and increasing ambient temperatures. Mild disease was reproduced experimentally by subcutaneous injection of Aeromonas hydrophila into apparently healthy frogs.

  20. Modular co-culture engineering, a new approach for metabolic engineering.

    Science.gov (United States)

    Zhang, Haoran; Wang, Xiaonan

    2016-09-01

    With the development of metabolic engineering, employment of a selected microbial host for accommodation of a designed biosynthetic pathway to produce a target compound has achieved tremendous success in the past several decades. Yet, increasing requirements for sophisticated microbial biosynthesis call for establishment and application of more advanced metabolic engineering methodologies. Recently, important progress has been made towards employing more than one engineered microbial strains to constitute synthetic co-cultures and modularizing the biosynthetic labor between the co-culture members in order to improve bioproduction performance. This emerging approach, referred to as modular co-culture engineering in this review, presents a valuable opportunity for expanding the scope of the broad field of metabolic engineering. We highlight representative research accomplishments using this approach, especially those utilizing metabolic engineering tools for microbial co-culture manipulation. Key benefits and major challenges associated with modular co-culture engineering are also presented and discussed.

  1. Triple co-culture cell model as an in vitro model for oral particulate vaccine systems

    DEFF Research Database (Denmark)

    Nielsen, Line Hagner; De Rossi, C.; Lehr, C-M.

    A triple co-culture cell model of Caco-2 cells, dendritic cells and macrophages (Figure 1) has previously been developed for studying intestinal permeability in a state of inflammation [1],[2]. The aim of this study was to investigate the applicability of this cell model for testing...... the model antigen ovalbumin was spray dried to obtain a particulate vaccine model system for testing in the cell model. The precursors were shown to form cubosomes when dispersed in aqueous medium, and was therefore used as the vaccine formulation for testing on the co-cultures. After 11 days, the TEER...... values of the co-cultures were found to be 860-1340 Ω∙cm2; the formulations were incubated with the co-cultures at this time point. From confocal microscopy images, it was observed that the THP-1 cells (macrophages) migrated into the overlying Caco-2 cell monolayer when the co-cultures were incubated...

  2. Aeromonas aquatica sp. nov., Aeromonas finlandiensis sp. nov. and Aeromonas lacus sp. nov. isolated from Finnish waters associated with cyanobacterial blooms.

    Science.gov (United States)

    Beaz-Hidalgo, R; Latif-Eugenín, F; Hossain, M J; Berg, K; Niemi, R M; Rapala, J; Lyra, C; Liles, M R; Figueras, M J

    2015-05-01

    Three groups of Aeromonas strains isolated from Finland lakes experiencing cyanobacterial blooms could not be assigned to any known species of this genus on the basis of 16S rRNA and rpoD gene sequences. The Multilocus Phylogenetic Analysis (MLPA) of the concatenated sequence of seven genes (gyrB, rpoD, recA, dnaJ, gyrA, dnaX and atpD; 4093bp) showed that the three groups of strains did not cluster with any known Aeromonas spp. and formed three independent lineages. This was confirmed by performing the analysis with their closest relatives using 15 genes (the latter 7 and cpn60, dnaK, gltA, mdh, radA, rpoB, tsf, zipA; 8751bp). Furthermore, ANI results between the genomes of the type strains of the three potential new species and those of their close relatives were all Aeromonas species on the basis of several phenotypic characters. This polyphasic study revealed that the 3 groups of strains represent 3 novel Aeromonas species for which the names Aeromonas aquatica sp. nov. (type strain AE235T=CECT 8025T=LMG 26712T), Aeromonas finlandiensis sp. nov. (type strain 4287DT=CECT 8028T=LMG 26709T) and Aeromonas lacus sp. nov. (type strain AE122T=CECT 8024T=LMG 26710T) are proposed.

  3. 西伯利亚鲟致病性嗜水气单胞菌外膜蛋白及其抗原性分析%Antigenicity of outer membrane proteins of pathogenic Aeromonas hydrophila isolated from Siberian sturgeon Acipenser baerii

    Institute of Scientific and Technical Information of China (English)

    姜有声; 杨倩; 李圆圆; 杨先乐; 吕利群; 邱军强

    2012-01-01

    A pathogenic strain Aeromonas hydrophila was isolated from Siberian sturgeon ( Acipenser baerii ) suffering with bacterial septicaemia. In this experiment, OMPs of four strains of Aeromonas were obtained by Sarkosyl method combining super centrifugal purification. SDS-PAGE showed that the strain had 14 membrane proteins with molecular weights of from 26 kDa to 120 kDa. OMPs of the pathogencial and unpathogencial Aeromonas hydrophila, Aeromonas caviae and Aeromonas sobria were analyzed and compared by using SDS-PAGE. The results showed that molecular weights focused on from 26 kDa to 120 kDa. After SDS-PAGE, in the Western-blotting with rabbit antiserum against Aeromonas hydrophila isolated from Siberian sturgeon, only 6 OMPs of the strain with molecular weights at 75 kDa, 52 kDa,43 kDa,40 kDa,34 kDa and 28kDa were detected. It showed that the 6 OMPs had some antigenicity. And the other 3 strains of Aeromonas all had positive reaction. The band of 28kDa and 34kDa had visible positive reaction in all 4 Aeromonas strains, the band of 43kDa and 75 kDa had positive reaction in some Aeromonas strains.%采用十二烷基肌氨酸钠(Sarkosyl)法提取西伯利亚鲟嗜水气单胞菌(Aeromonas hydrophila)外膜蛋白,电泳显示所提取的主要外膜蛋白分子量为26~120 kDa;为比较该菌株与气单胞菌菌属其他细菌外膜蛋白组分及抗原性异同,以致病性豚鼠气单胞菌(A.caviae)、温和气单胞菌(A.sobria)和无致病力的嗜水气单胞菌为对照,电泳图谱显示4种气单胞菌外膜蛋白的分子量主要集中在26~120 kDa之间;利用抗西伯利亚鲟嗜水气单胞菌血清的免疫印迹试验表明该菌株外膜蛋白中分子量为75 kDa、52 kDa、43 kDa、40 kDa、34 kDa、28 kDa的蛋白条带呈现阳性反应,其他3种气单胞菌外膜蛋白中均有与该抗血清反应的条带,且分子量为28 kDa、34 kDa的反应条带为4株菌共有;43 kDa与75 kDa反应条带为部分菌株共有.为进一步筛选和

  4. Complete genome sequence of Aeromonas hydrophila AL06-06

    Science.gov (United States)

    Aeromonas hydrophila occurs in freshwater environments and infects fish and mammals. In this work, we report the complete genome sequence of Aeromonas hydrophila AL06-06, which was isolated from diseased goldfish and is being used for comparative genomic studies with A. hydrophila strains causing ba...

  5. Draft genome sequence of Aeromonas hydrophila TN97-08

    Science.gov (United States)

    Aeromonas hydrophila is an opportunistic Gram-negative species causing disease in fish and mammals. The genus Aeromonas affects a variety of aquatic organisms and lives in diverse aquatic ecosystems (1). There are 39 A. hydrophila genomes currently available in GenBank. In the current study, we repo...

  6. OVERVIEW: DISINFECTION OF HELICOBACTER PYLORI AND AEROMONAS SPECIES

    Science.gov (United States)

    Helicobacter pylori and Aeromonas hydrophila are contaminants listed on the USEPA's 1998 Contaminant Candidate List (CCL).The sensitivity of H. pylori to chlorine and of Aeromonas spp. to inactivation by free chlorine, chloramine and ultraviolet (UV) was examined. Selective and...

  7. Chitin degradation and metabolism by virulent Aeromonas hydropila

    Science.gov (United States)

    Aeromonas hydrophila is the causal agent of motile Aeromonas septicemia (MAS) in catfish and other warm-water fishes. Severe outbreak of MAS caused by virulent A. hydrophila (vAh) was reported in 2009 in the Southeastern United States; the disease has since resulted in loss of millions of pounds of ...

  8. Validation of a Behavioral Ethogram for Assessing Postoperative Pain in Guinea Pigs (Cavia porcellus)

    OpenAIRE

    Dunbar, Misha L; David, Emily M; Aline, Marian R; Lofgren, Jennifer L.

    2016-01-01

    Although guinea pigs (Cavia porcellus) have been used in research for more than a century and remain the most prevalent USDA-covered species, little has been elucidated regarding the recognition of clinical pain or analgesic efficacy in this species. We sought to assess pain in guinea pigs by using newer, clinically relevant methods that have been validated in other rodent species: the behavioral ethogram and cageside proxy indicator. In this study, 10 male guinea pigs underwent electronic vo...

  9. Isolation of Aeromonas spp. from Food Products: Emerging Aeromonas Infections and Their Significance in Public Health.

    Science.gov (United States)

    Alhazmi, Mohammad Ismail

    2015-01-01

    Aeromonas spp. are opportunistic pathogens causing a broad spectrum of human illnesses like gastroenteritis, chronic diarrhea, wound infections, peritonitis, urinary tract infections, and septicemia. Their ability to grow in foods stored in a refrigerator poses a substantial threat for human consumption. We investigated the prevalence of Aeromonas from commercial food products across Riyadh, Saudi Arabia. A total of 250 samples were randomly collected and processed for the isolation and identification of Aeromonas by morphological and biochemical means and for their identification by PCR. A total of 102 strains of Aeromonas were isolated, including 47% from raw meat samples, 34% from raw fish samples, and 18.6% from milk and dairy products; 56.8% were identified as A. hydrophila and 43.1% as A. sobria. Antibiotic susceptibility tests done revealed 100% sensitivity to chloramphenicol, colistin, ciprofloxacin, and nitrofurantoin. 16S rDNA PCR revealed the presence of the 953 bp fragment in all the strains. The present investigation suggested the occurrences of A. sobria and A. hydrophila in human consumable stored and refrigerated foods.

  10. Isolation and identification of Aeromonas and analysis for drug resistance from diarrhea samples in Shanghai Pudong New Area%浦东新区腹泻样本中气单胞菌的分离鉴定及耐药性分析

    Institute of Scientific and Technical Information of China (English)

    王闻卿; 赵冰; 詹铭

    2011-01-01

    Objective: To find out the incidence of Aeromonas infection and the drug resistance from diarrhea cases in Shanghai Pudong New Area. Methods: Detection Aeromonas in feces specimens from 1245 diarrhea samples in 2 monitoring sites; the positive strains were tested for sensitivity to 10 kinds of antibiotics by K - B method. Results: 93 stains(7.47% ) of Aeromonas were isolated, including 26 strains of Aeromonas sobria,63 strains of Aeromonas hydrophila and 4 strains of Aeromonas caviae. The detection rate of infection during June to October revealed higher prevelence rate. Antimicrobial susceptibility tests showed more than 80% strains were susceptible to Gentamicin, Ciprofloxacin, Amikacin, Cefotaxime while resistant to Ampicillin, Trimethoprim - sulfamethoxazole, Cefalotin, Nalidixic acid at deferent levels. Conclusion: Infection rate of Aeromonas varied in season, having close relationship with diarrhea in summer and autumn. Continuous Monitoring to Aeromonas and its drug resistance has guiding significance for prevention and treatment of diarrhea disease.%目的:了解上海浦东新区感染性腹泻病人气单胞菌的感染及耐药情况,为相关防控策略的制定提供科学依据.方法:对2010年上海市浦东新区2个监测点,1245件急性腹泻样本进行气单胞菌检测,阳性菌进行药物敏感试验.结果:从1245件标本中检出气单胞菌93株,阳性率为7.47%,其中嗜水气单胞63株,温和气单胞26株,豚鼠气单胞4株.6月-8月份为高峰期.药敏试验结果:80%以上菌株对庆大,环丙沙星,阿米卡星、头孢噻肟敏感,对氨苄西林,复方新诺明、四环素、头孢噻吩、萘啶酸不同程度耐药.结论:气单胞菌感染存在季节性差异,与夏秋季腹泻关系密切.持续对气单胞菌及其耐药性的监测对细菌性腹泻病的预防与治疗具有指导意义.

  11. Insight into the mobilome of Aeromonas strains

    Directory of Open Access Journals (Sweden)

    Marta ePiotrowska

    2015-05-01

    Full Text Available The mobilome is a pool of genes located within mobile genetic elements (MGE, such as plasmids, IS elements, transposons, genomic/pathogenicity islands, and integron-associated gene cassettes. These genes are often referred to as flexible and may encode virulence factors, toxic compounds as well as resistance to antibiotics. The phenomenon of MGE transfer between bacteria, known as horizontal gene transfer (HGT, is well documented. The genes present on MGE are subject to continuous processes of evolution and environmental changes, largely induced or significantly accelerated by man. For bacteria, the only chance of survival in an environment contaminated with toxic chemicals, heavy metals and antibiotics is the acquisition of genes providing the ability to survive in such conditions. The process of acquiring and spreading antibiotic resistance genes (ARG is of particular significance, as it is important for the health of humans and animals. Therefore, it is important to thoroughly study the mobilome of Aeromonas spp. that is widely distributed in various environments, causing many diseases in fishes and humans. This review discusses the recently published information on MGE prevalent in Aeromonas spp. with special emphasis on plasmids belonging to different incompatibility groups, i.e., IncA/C, IncU, IncQ, IncF, IncI and ColE-type. The vast majority of plasmids carry a number of different transposons (Tn3, Tn21, Tn1213, Tn1721, Tn4401, the 1st, 2nd or 3rd class of integrons, IS elements (e.g. IS26, ISPa12, ISPa13, ISKpn8, ISKpn6 and encode determinants such as antibiotic and mercury resistance genes, as well as virulence factors. Although the actual role of Aeromonas spp. as a human pathogen remains controversial, species of this genus may pose a serious risk to human health. This is due to the considerable potential of their mobilome, particularly in terms of antibiotic resistance and the possibility of the horizontal transfer of resistance

  12. AEROMONAS SALMONICIDA INFECTION IN VACCINATED RAINBOW TROUT

    DEFF Research Database (Denmark)

    Chettri, Jiwan Kumar; Skov, Jakob; Mohammad, Rezkar Jaafar

    In vivo testing of any candidate vaccine is influenced by the choice of challenge method and the external environmental conditions. In the present study, a comparative challenge study was performed to evaluate the efficacy of different vaccines against the bacterial pathogen Aeromonas salmonicida...... in saltwater condition produced less mortality compared to freshwater, probably due to the growth inhibition of A. salmonicida in saline condition which was also verified by in vitro assay. One of the experimental vaccines emulsified in water in oil adjuvant showed a protection comparable...... effects due to oil adjuvants, and the importance of choice of challenge methods used will be discussed....

  13. Depot-dependent effects of adipose tissue explants on co-cultured hepatocytes

    DEFF Research Database (Denmark)

    Du, Zhen-Yu; Ma, Tao; Lock, Erik-Jan;

    2011-01-01

    We have developed an in vitro hepatocyte-adipose tissue explant (ATE) co-culture model enabling examination of the effect of visceral and subcutaneous adipose tissues on primary rat hepatocytes. Initial analyses of inflammatory marker genes were performed in fractionated epididymal or inguinal...... levels of IL-6, TNF-a and PGE(2) in the media from inguinal ATEs co-cultured with primary rat hepatocytes were higher than that in the media from epididymal ATEs co-cultured with hepatocytes, although the significant difference was only seen in PGE(2). Lipolysis, measured as glycerol release, was similar...... in the ATEs isolated from inguinal and epididymal adipose tissues when cultured alone, but the glycerol release was higher in the ATEs isolated from epididymal than from inguinal adipose tissue when co-cultured with hepatocytes. Compared to epididymal ATEs, the ATEs from inguinal adipose tissue elicited...

  14. Production of probiotic fresh white cheese using co-culture with Streptococcus thermophilus

    OpenAIRE

    Oktay Yerlikaya; Elif Ozer

    2014-01-01

    In this research, the probiotic Streptococcus thermophilus was inoculated into milk as co-culture to produce probiotic cheese. The effects of using Streptococcus thermophilus with other probiotic bacteria on cheese composition, and microbiological viability during 28 days of storage were investigated. Sensorial properties were determined only at 1st and 28th days of storage. The results showed that the use of Streptococcus thermophilus as co-culture in probiotic cheese production did not affe...

  15. Streamlining Gene Expression Analysis: Integration of Co-Culture and mRNA Purification

    OpenAIRE

    Berry, Scott M; Singh, Chandresh; Lang, Jessica D.; Strotman, Lindsay N.; Alarid, Elaine T.; Beebe, David J.

    2014-01-01

    Co-culture of multiple cell types within a single device enables the study of paracrine signaling events. However, extracting gene expression endpoints from co-culture experiments is laborious, due in part to pre-PCR processing of the sample (i.e., post-culture cell sorting, nucleic acid purification). Also, significant loss of nucleic acid may occur during these steps, especially with microfluidic cell culture where lysate volumes are small and difficult to access. Here, we describe an integ...

  16. Generation of a patterned co-culture system composed of adherent cells and immobilized nonadherent cells.

    Science.gov (United States)

    Yamazoe, Hironori; Ichikawa, Takashi; Hagihara, Yoshihisa; Iwasaki, Yasuhiko

    2016-02-01

    Patterned co-culture is a promising technique used for fundamental investigation of cell-cell communication and tissue engineering approaches. However, conventional methods are inapplicable to nonadherent cells. In this study, we aimed to establish a patterned co-culture system composed of adherent and nonadherent cells. Nonadherent cells were immobilized on a substrate using a cell membrane anchoring reagent conjugated to a protein, in order to incorporate them into the co-culture system. Cross-linked albumin film, which has unique surface properties capable of regulating protein adsorption, was used to control their spatial localization. The utility of our approach was demonstrated through the fabrication of a patterned co-culture consisting of micropatterned neuroblastoma cells surrounded by immobilized myeloid cells. Furthermore, we also created a co-culture system composed of cancer cells and immobilized monocytes. We observed that monocytes enhanced the drug sensitivity of cancer cells and its influence was limited to cancer cells located near the monocytes. Therefore, the incorporation of nonadherent cells into a patterned co-culture system is useful for creating culture systems containing immune cells, as well as investigating the influence of these immune cells on cancer drug sensitivity. Various methods have been proposed for creating patterned co-culture systems, in which multiple cell types are attached to a substrate with a desired pattern. However, conventional methods, including our previous report published in Acta Biomaterialia (2010, 6, 526-533), are unsuitable for nonadherent cells. Here, we developed a novel method that incorporates nonadherent cells into the co-culture system, which allows us to precisely manipulate and study microenvironments containing nonadherent and adherent cells. Using this technique, we demonstrated that monocytes (nonadherent cells) could enhance the drug sensitivity of cancer cells and that their influence had a

  17. Decolorization of synthetic brilliant green carpet industry dye through fungal co-culture technology.

    Science.gov (United States)

    Kumari, Simpal; Naraian, Ram

    2016-09-15

    Aim of the present study was to evaluate the efficiency of fungal co-culture for the decolorization of synthetic brilliant green carpet industry dye. For this purpose two lignocellulolytic fungi Pleurotus florida (PF) and Rhizoctonia solani (RS) were employed. The study includes determination of enzyme profiles (laccase and peroxidase), dye decolorization efficiency of co-culture and crude enzyme extracts. Both fungi produced laccase and Mn peroxidase and successfully decolorized solutions of different concentrations (2.0, 4.0, 6.0, & 8.0(w/v) of dye. The co-culture resulted highest 98.54% dye decolorization at 2% (w/v) of dye as compared to monocultures (82.12% with PF and 68.89% with RS) during 12 days of submerged fermentation. The lower levels of dyes were rapidly decolorized, while higher levels in slow order as 87.67% decolorization of 8% dye. The promising achievement of the study was remarkable decolorizing efficiency of co-culture over monocultures. The direct treatment of the mono and co-culture enzyme extracts to dye also influenced remarkable. The highest enzymatic decolorization was through combined (PF and RS) extracts, while lesser by monoculture extracts. Based on the observations and potentiality of co-culture technology; further it can be exploited for the bioremediation of areas contaminated with hazardous environmental pollutants including textile and other industry effluents.

  18. Advances in tissue engineering through stem cell-based co-culture.

    Science.gov (United States)

    Paschos, Nikolaos K; Brown, Wendy E; Eswaramoorthy, Rajalakshmanan; Hu, Jerry C; Athanasiou, Kyriacos A

    2015-05-01

    Stem cells are the future in tissue engineering and regeneration. In a co-culture, stem cells not only provide a target cell source with multipotent differentiation capacity, but can also act as assisting cells that promote tissue homeostasis, metabolism, growth and repair. Their incorporation into co-culture systems seems to be important in the creation of complex tissues or organs. In this review, critical aspects of stem cell use in co-culture systems are discussed. Direct and indirect co-culture methodologies used in tissue engineering are described, along with various characteristics of cellular interactions in these systems. Direct cell-cell contact, cell-extracellular matrix interaction and signalling via soluble factors are presented. The advantages of stem cell co-culture strategies and their applications in tissue engineering and regenerative medicine are portrayed through specific examples for several tissues, including orthopaedic soft tissues, bone, heart, vasculature, lung, kidney, liver and nerve. A concise review of the progress and the lessons learned are provided, with a focus on recent developments and their implications. It is hoped that knowledge developed from one tissue can be translated to other tissues. Finally, we address challenges in tissue engineering and regenerative medicine that can potentially be overcome via employing strategies for stem cell co-culture use. Copyright © 2014 John Wiley & Sons, Ltd.

  19. Differential media for quantitative recovery of waterborne Aeromonas hydrophila.

    Science.gov (United States)

    Handfield, M; Simard, P; Letarte, R

    1996-09-01

    Because of the ubiquity of Aeromonas spp., their prevalence in drinking water, and the increasing number of reports on Aeromonas sp.-related infections, a standard method for routine and quantitative recovery had to be defined. On the basis of a survey of 10 media for recovery analysis and subsequent differentiation assays in mixed cultures, we conclude that ampicillin-dextrin agar performed the best for the recovery of Aeromonas spp. in drinking water and the differentiation by simple criteria of that genus from other common waterborne bacteria.

  20. Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Fan, Ping, E-mail: fanpinggoodluck@163.com [Department of Rheumatism and Immunity, The First Affiliated Hospital Xi' an Jiaotong University School of Medicine, Xi' an, Shaanxi 710061 (China); He, Lan; Pu, Dan; Lv, Xiaohong; Zhou, Wenxu; Sun, Yining; Hu, Nan [Department of Rheumatism and Immunity, The First Affiliated Hospital Xi' an Jiaotong University School of Medicine, Xi' an, Shaanxi 710061 (China)

    2011-01-21

    Research highlights: {yields} The proliferation of dramatic increased by co-cultured with Sertoli cells. {yields} VEGF receptor-2 expression of ECs was up-regulated by co-cultured with Sertoli cells. {yields} The MHC expression of ECs induced by INF-{gamma} and IL-6, IL-8 and sICAM induced by TNF-{alpha} decreased respectively after co-cultured with Sertoli cells. {yields} ECs co-cultured with Sertoli cells also didn't increase the stimulation index of spleen lymphocytes. -- Abstract: The major problem of the application of endothelial cells (ECs) in transplantation is the lack of proliferation and their immunogenicity. In this study, we co-cultured ECs with Sertoli cells to monitor whether Sertoli cells can influence the proliferation and immunogenicity of co-cultured ECs. Sertoli cells were isolated from adult testicular tissue. ECs were divided into the control group and the experimental group, which included three sub-groups co-cultured with 1 x 10{sup 3}, 1 x 10{sup 4} or 1 x 10{sup 5} cell/ml of Sertoli cells. The growth and proliferation of ECs were observed microscopically, and the expression of vascular endothelial growth factor (VEGF) receptor-2 (KDR) was examined by Western blotting. In another experiment, ECs were divided into the control group, the single culture group and the co-culture group with the optimal concentration of Sertoli cells. After INF-{gamma} and TNF-{alpha} were added to the culture medium, MHC II antigen expression was detected by immunofluorescence staining and western blotting; interleukin (IL)-6, IL-8 and soluble intercellular adhesion molecule (sICAM) were measured in the culture medium by ELISA. We demonstrated that 1 x 10{sup 4} cell/ml Sertoli cells promoted the proliferation of co-cultured ECs more dramatically than that in other groups (P < 0.05). Western blotting showed that 1 x 10{sup 4} cell/ml of the Sertoli cells was most effective in the up-regulation of KDR expression in the co-cultured ECs (P < 0.05). Sertoli

  1. Studies concerning the antigen affinities of Aeromonas punctata strains

    National Research Council Canada - National Science Library

    Krug, S

    1982-01-01

    On the basis of 90 determined Aeromonas punctata strains from different fish species serological cross reactions were carried out with the aid of agglutination and precipitation to determine antigen...

  2. Histopathological findings in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with 3 different Aeromonas species.

    Science.gov (United States)

    Zepeda-Velázquez, Andrea Paloma; Vega-Sánchez, Vicente; Salgado-Miranda, Celene; Soriano-Vargas, Edgardo

    2015-07-01

    This study describes the macroscopic and microscopic lesions in farmed rainbow trout (Oncorhynchus mykiss) naturally infected with genetically identified Aeromonas salmonicida, A. hydrophila, and A. veronii species. The genus Aeromonas includes bacteria that naturally inhabit both waterways and organisms. At least 27 Aeromonas species have been identified to date, some of which can cause significant economic losses in aquaculture. As up to 68.8% of Aeromonas isolates may be misidentified in routine biochemical and phenotypic tests, however, reported cases of Aeromonas infection in fish may be wrongly identified. Our findings confirmed that the 3 Aeromonas species studied are associated with septicemia and dermal lesions in rainbow trout.

  3. Post-traumatic osteomyelitis due to aeromonas species

    Directory of Open Access Journals (Sweden)

    Gunasekaran L

    2009-01-01

    Full Text Available We report a case of Aeromonas osteomyelitis due to injury in a sewage worker. He presented with cellulitis of the left foot. Radiographs showed evidence of osteomyelitis involving the head and neck of the fifth metatarsal. Aeromonas species was isolated from the tissue and swab from the foot. The head and neck of the fifth metatarsal were excised and the patient improved on 4 weeks of intravenous meropenem followed by 4 weeks of oral clindamycin and ciprofloxacin.

  4. Emerging Aeromonas Species Infections and Their Significance in Public Health

    OpenAIRE

    Igbinosa, Isoken H.; Ehimario U. Igumbor; Farhad Aghdasi; Mvuyo Tom; Anthony I. Okoh

    2012-01-01

    Aeromonas species are ubiquitous bacteria in terrestrial and aquatic milieus. They are becoming renowned as enteric pathogens of serious public health concern as they acquire a number of virulence determinants that are linked with human diseases, such as gastroenteritis, soft-tissue, muscle infections, septicemia, and skin diseases. Proper sanitary procedures are essential in the prevention of the spread of Aeromonas infections. Oral fluid electrolyte substitution is employed in the preventio...

  5. Plasmid-cured Chlamydia caviae activates TLR2-dependent signaling and retains virulence in the guinea pig model of genital tract infection.

    Directory of Open Access Journals (Sweden)

    Lauren C Frazer

    Full Text Available Loss of the conserved "cryptic" plasmid from C. trachomatis and C. muridarum is pleiotropic, resulting in reduced innate inflammatory activation via TLR2, glycogen accumulation and infectivity. The more genetically distant C. caviae GPIC is a natural pathogen of guinea pigs and induces upper genital tract pathology when inoculated intravaginally, modeling human disease. To examine the contribution of pCpGP1 to C. caviae pathogenesis, a cured derivative of GPIC, strain CC13, was derived and evaluated in vitro and in vivo. Transcriptional profiling of CC13 revealed only partial conservation of previously identified plasmid-responsive chromosomal loci (PRCL in C. caviae. However, 2-deoxyglucose (2DG treatment of GPIC and CC13 resulted in reduced transcription of all identified PRCL, including glgA, indicating the presence of a plasmid-independent glucose response in this species. In contrast to plasmid-cured C. muridarum and C. trachomatis, plasmid-cured C. caviae strain CC13 signaled via TLR2 in vitro and elicited cytokine production in vivo similar to wild-type C. caviae. Furthermore, inflammatory pathology induced by infection of guinea pigs with CC13 was similar to that induced by GPIC, although we observed more rapid resolution of CC13 infection in estrogen-treated guinea pigs. These data indicate that either the plasmid is not involved in expression or regulation of virulence in C. caviae or that redundant effectors prevent these phenotypic changes from being observed in C. caviae plasmid-cured strains.

  6. Aeromonas hydrophila Sepsis Mimicking Vibrio vulnificus Infection.

    Science.gov (United States)

    Park, Se Young; Nam, Hyun Min; Park, Kun; Park, Seok Don

    2011-09-01

    Aeromonas hydrophila is a facultatively anaerobic, asporogenous gram-negative rod that has often been regarded as an opportunistic pathogen in hosts with impairment of a local or general defense mechanism. A 68-year-old alcoholic woman presented with shock and gangrene on the right arm. At first, her clinical presentations were severe painful erythematous swelling that worsened within a few hours with development of gangrene, edema, and blisters. Bullous fluid and blood cultures yielded A. hydrophila. Histopathological findings of sections obtained from the vesicle revealed subepidermal vesicles; necrosis of the epidermis, papillary dermis, and subcutaneous fat; and massive hemorrhage in the subcutis. Despite all efforts to save the patient, she died 8 hours after admission. Clinical features of A. hydrophila sepsis resemble those of Vibrio vulnificus sepsis. Therefore, in addition to the case report, we compared the cultural, biochemical, and morphological differences between A. hydrophila and V. vulnificus for facilitation of early and accurate identification of the causative agent.

  7. Investigation of Interspecies Interactions within Marine Micromonosporaceae Using an Improved Co-Culture Approach

    Directory of Open Access Journals (Sweden)

    Navid Adnani

    2015-09-01

    Full Text Available With respect to bacterial natural products, a significant outcome of the genomic era was that the biosynthetic potential in many microorganisms surpassed the number of compounds isolated under standard laboratory growth conditions, particularly among certain members in the phylum Actinobacteria. Our group, as well as others, investigated interspecies interactions, via co-culture, as a technique to coax bacteria to produce novel natural products. While co-culture provides new opportunities, challenges exist and questions surrounding these methods remain unanswered. In marine bacteria, for example, how prevalent are interspecies interactions and how commonly do interactions result in novel natural products? In an attempt to begin to answer basic questions surrounding co-culture of marine microorganisms, we have tested both antibiotic activity-based and LC/MS-based methods to evaluate Micromonosporaceae secondary metabolite production in co-culture. Overall, our investigation of 65 Micromonosporaceae led to the identification of 12 Micromonosporaceae across three genera that produced unique metabolites in co-culture. Our results suggest that interspecies interactions were prevalent between marine Micromonosporaceae and marine mycolic acid-containing bacteria. Furthermore, our approach highlights a sensitive and rapid method for investigating interspecies interactions in search of novel antibiotics, secondary metabolites, and genes.

  8. Co-culture systems and technologies: taking synthetic biology to the next level.

    Science.gov (United States)

    Goers, Lisa; Freemont, Paul; Polizzi, Karen M

    2014-07-06

    Co-culture techniques find myriad applications in biology for studying natural or synthetic interactions between cell populations. Such techniques are of great importance in synthetic biology, as multi-species cell consortia and other natural or synthetic ecology systems are widely seen to hold enormous potential for foundational research as well as novel industrial, medical and environmental applications with many proof-of-principle studies in recent years. What is needed for co-cultures to fulfil their potential? Cell-cell interactions in co-cultures are strongly influenced by the extracellular environment, which is determined by the experimental set-up, which therefore needs to be given careful consideration. An overview of existing experimental and theoretical co-culture set-ups in synthetic biology and adjacent fields is given here, and challenges and opportunities involved in such experiments are discussed. Greater focus on foundational technology developments for co-cultures is needed for many synthetic biology systems to realize their potential in both applications and answering biological questions.

  9. "Co-culture Engineering" for Enhanced Phytoremediation of Metal Contaminated Soils

    Institute of Scientific and Technical Information of China (English)

    NI Cai-Ying; SHI Ji-Yan; LUO Yong-Ming; CHEN Ying-Xu

    2004-01-01

    A co-culture of two plant materials, Astragalus sinicus L., a leguminous plant with concomitant nodules, and Elsholtzia splendens Naki-a Cu accumulator, along with treatments of a chelating agent (EDTA), root excretions (citric acid), and a control with E. splendens only were used to compare the mobility of heavy metals in chelating agents with a co-culture and to determine the potential for co-culture phytoremediation in heavy metal contaminated soils. The root uptake for Cu, Zn, and Pb in all treatments was significantly greater (P < 0.05) than that of the control treatment. However with translocation in the shoots, only Cu, Zn, and Pb in plants grown with the EDTA treatment and Zn in plants cocropped with the A. sinicus treatment increased significantly (P < 0.05). In addition, when a co-culture in soils with heavy and moderate contamination was compared, for roots in moderately contaminated soils only Zn concentration was significantly less (P < 0.05) than that of heavily contaminated soils, however, Cu, Zn, and Pb concentrations of shoots were all significantly lower (P < 0.05). Overall, this "co-culture engineering" could be as effective as or even more effective than chelating agents, thereby preventing plant metal toxicity and metal leaching in soils as was usually observed in chelate-enhanced phytoremediation.

  10. Potential of fungal co-culturing for accelerated biodegradation of petroleum hydrocarbons in soil.

    Science.gov (United States)

    Yanto, Dede Heri Yuli; Tachibana, Sanro

    2014-08-15

    The potential of fungal co-culture of the filamentous Pestalotiopsis sp. NG007 with four different basidiomycetes--Trametes versicolor U97, Pleurotus ostreatus PL1, Cerena sp. F0607, and Polyporus sp. S133--for accelerating biodegradation of petroleum hydrocarbons (PHCs) was studied using three different physicochemical characteristic PHCs in soil. All the combinations showed a mutual intermingling mycelial interaction on the agar plates. However, only NG007/S133 (50/50) exhibited an optimum growth rate and enzymatic activities that supported the degradation of asphalt in soil. The co-culture also degraded all fractions at even higher concentrations of the different PHCs. In addition, asphaltene, which is a difficult fraction for a single microorganism to degrade, was markedly degraded by the co-culture, which indicated that the simultaneous biodegradation of aliphatic, aromatic, resin, and asphaltene fractions had occurred in the co-culture. An examination of in-vitro degradation by the crude enzymes and the retrieval fungal culture from the soil after the experiment confirmed the accelerated biodegradation due to enhanced enzyme activities in the co-culture. The addition of piperonyl butoxide or AgNO3 inhibited biodegradation by 81-99%, which demonstrated the important role of P450 monooxygenases and/or dioxygenases in the initial degradation of the aliphatic and aromatic fractions in PHCs.

  11. Garlic exerts allelopathic effects on pepper physiology in a hydroponic co-culture system.

    Science.gov (United States)

    Ding, Haiyan; Cheng, Zhihui; Liu, Menglong; Hayat, Sikandar; Feng, Han

    2016-05-15

    A hydroponic co-culture system was adopted to determine the allelopathic potential of garlic on the growth of pepper plants. Different numbers of garlic plants (0, 2, 4, 8 and 12) were hydroponically co-cultured with two pepper plants to investigate allelopathic effects on the growth attributes and antioxidative defense system of the test pepper plants. The responses of the pepper plants depended on the number of garlic plants included in the co-culture system, indicating an association of pepper growth with the garlic root exudate concentration. When grown at a pepper/garlic ratio of 1:1 or 1:2, the pepper plant height, chlorophyll content, and peroxidase (POD), catalase (CAT) and phenylalanine ammonia-lyase (PAL) activities were significantly increased after 30 days of co-culture; in contrast, reduction in methane dicarboxylic aldehyde (MDA) content was observed. However, when the pepper/garlic ratio was 1:4 or higher, these morphological indices and protective enzyme activities were significantly inhibited, whereas MDA levels in the pepper leaves were significantly increased due to severe membrane lipid peroxidation. The results indicate that although low concentrations of garlic root exudates appear to induce protective enzyme systems and promote pepper growth, high concentrations have deleterious effects. These findings suggest that further investigations should optimize the co-culture pepper/garlic ratio to reduce continuous cropping obstacles in pepper production.

  12. Application of cell co-culture system to study fat and muscle cells.

    Science.gov (United States)

    Pandurangan, Muthuraman; Hwang, Inho

    2014-09-01

    Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.

  13. Garlic exerts allelopathic effects on pepper physiology in a hydroponic co-culture system

    Directory of Open Access Journals (Sweden)

    Haiyan Ding

    2016-05-01

    Full Text Available A hydroponic co-culture system was adopted to determine the allelopathic potential of garlic on the growth of pepper plants. Different numbers of garlic plants (0, 2, 4, 8 and 12 were hydroponically co-cultured with two pepper plants to investigate allelopathic effects on the growth attributes and antioxidative defense system of the test pepper plants. The responses of the pepper plants depended on the number of garlic plants included in the co-culture system, indicating an association of pepper growth with the garlic root exudate concentration. When grown at a pepper/garlic ratio of 1:1 or 1:2, the pepper plant height, chlorophyll content, and peroxidase (POD, catalase (CAT and phenylalanine ammonia-lyase (PAL activities were significantly increased after 30 days of co-culture; in contrast, reduction in methane dicarboxylic aldehyde (MDA content was observed. However, when the pepper/garlic ratio was 1:4 or higher, these morphological indices and protective enzyme activities were significantly inhibited, whereas MDA levels in the pepper leaves were significantly increased due to severe membrane lipid peroxidation. The results indicate that although low concentrations of garlic root exudates appear to induce protective enzyme systems and promote pepper growth, high concentrations have deleterious effects. These findings suggest that further investigations should optimize the co-culture pepper/garlic ratio to reduce continuous cropping obstacles in pepper production.

  14. Peritonitis bacteriana espontánea y bacteriemia por Aeromonas hydrophila Spontaneous bacterial peritonitis and bacteraemia due to Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    J. J. García-Irure

    2003-12-01

    Full Text Available La peritonitis bacteriana es una complicación frecuente en los pacientes cirróticos. De los gérmenes etiológicos, Aeromonas es muy poco frecuente. Presentamos un caso clínico de un paciente cirrótico con peritonitis y bacteriemia por Aeromonas hydrophila. Este microorganismo es un bacilo gramnegativo de la familia Vibrionaceae. En el ser humano produce gastroenteritis con mucha frecuencia. De forma excepcional puede dar origen a infecciones extraintestinales, sobre todo en pacientes inmunodeprimidos. La peritonitis por Aeromonas se ha descrito en asociación a peritonitis bacteriana espontánea en pacientes cirróticos, en pacientes en diálisis peritoneal y en casos de peritonitis secundaria a perforación intestinal.Bacterial peritonitis is a frequent complication in cirrhotic patients. Amongst the aetiological germs, Aeromonas is very infrequent. We present the clinical case of a cirrhotic patient with peritonitis and bacteraemia due to Aeromonas hydrophila. This micro-organism is a Gram-negative bacillus of the Vibrionaceae family. It very frequently produces gastroenteritis in the human being. Exceptionally it can give rise to extraintestinal infections, especially in immunodepressed patients. Peritonitis due to Aeromonas has been described in association with spontaneous bacterial peritonitis in cirrhotic patients, in patients on peritoneal dialysis and in cases of peritonitis secondary to intestinal perforation.

  15. Co-culture of astrocytes with neurons from injured brain A time-dependent dichotomy

    Institute of Scientific and Technical Information of China (English)

    Xiaojing Xu; Min Wang; Jing Liu; Jingya Lv; Yanan Hu; Huanxiang Zhang

    2011-01-01

    As supportive cells for neuronal growth and development, much effort has been devoted to the role of astrocytes in the normal state. However, the effect of the astrocytes after injury remains elusive. In the present study, neurons isolated from the subventricular zone of injured neonatal rat brains were co-cultured with astrocytes. After 6 days, these astrocytes showed a mature neuron-like appearance and the number of survivingneurons, primary dendrites and total branches was significantly higher than those at 3 days. The neurons began to shrink at 9 days after co-culture with shorter and thinner processes and the number of primary dendrites and total branches was significantly reduced. These experimental findings indicate that astrocytes in the injured brain promote the development of neurons in the early stages of co-culture while these cells reversely inhibit neuronal growth and development at the later states.

  16. A comparative analysis of the intestinal metagenomes present in guinea pigs (Cavia porcellus) and humans (Homo sapiens)

    DEFF Research Database (Denmark)

    Hildebrand, Falk; Ebersbach, Tine; Nielsen, Henrik Bjørn

    2012-01-01

    Background: Guinea pig (Cavia porcellus) is an important model for human intestinal research. We have characterized the faecal microbiota of 60 guinea pigs using Illumina shotgun metagenomics, and used this data to compile a gene catalogue of its prevalent microbiota. Subsequently, we compared th...

  17. Draft Genome Sequence of Streptococcus caviae Strain Cavy grass 6T, Isolated from Domesticated Guinea Pig Fecal Samples

    Science.gov (United States)

    Palakawong Na Ayudthaya, Susakul; Marshall, Ian P. G.; Schreiber, Lars

    2017-01-01

    ABSTRACT Streptococcus caviae strain Cavy grass 6T, isolated from fecal samples of pet guinea pigs, can metabolize a range of plant mono- and disaccharides, as well as polymeric carbohydrates. Here, we report the draft genome sequence of this strain, which comprises 2.11 Mb. PMID:28360157

  18. Detection of Chlamydophila caviae and Streptococcus equi subsp. zooepidemicus in horses with signs of rhinitis and conjunctivitis.

    Science.gov (United States)

    Gaede, Wolfgang; Reckling, Karl-Friedrich; Schliephake, Anette; Missal, Dirk; Hotzel, Helmut; Sachse, Konrad

    2010-05-19

    At a stud farm of Trakehner horses, all 33 foals of a birth cohort developed conjunctivitis and serous to muco-purulent rhinitis, and 7 older horses showed recurrent signs of conjunctivitis. Examination of nasal and conjunctival swabs by bacterial and cell culture, as well as real-time PCR, ArrayTube microarray analysis and DNA sequencing led to the identification of Chlamydophila (C.) caviae (first description in horses) and Streptococcus (S.) equi subsp. zooepidemicus. We presume a synergistic effect associated with these two agents by hypothesising that primary lesions were set by C. caviae and subsequently aggravated by Streptococcus equi subsp. zooepidemicus. Indications supporting this assumption include (i) the conjunctivitis caused by mono-infection with C. caviae, (ii) recurrent clinical symptoms in the affected animals, and (iii) the absence of a sustained clinical effect of antibiotic therapy with trimethoprim-sulfonamide, enrofloxacin and amoxicillin. The detection of C. caviae in horses raises questions about the significance and natural host range of this agent.

  19. Ectoparasites of the critically endangered insular cavy, Cavia intermedia (Rodentia: Caviidae, southern Brazil

    Directory of Open Access Journals (Sweden)

    André Luis Regolin

    2015-04-01

    Full Text Available Cavia intermedia is a rodent species critically endangered and is found only on a 10 hectare island off the southern Brazilian coast. To identify the ectoparasites of C. intermedia, 27 specimens (14 males and 13 females, representing approximately 65% of the estimated total population, were captured and examined. A total of 1336 chewing lice of two species were collected: Gliricola lindolphoi (Amblycera: Gyropidae and Trimenopon hispidum (Amblycera: Trimenoponidae. In addition, chiggers Arisocerus hertigi (Acari: Trombiculidae and Eutrombicula sp. (Acari: Trombiculidae were collected from the ears of all captured animals. This low species richness compared to those for other Cavia species is expected for island mammals. Although the results presented here are not conclusive about the relationship between C. intermedia and ectoparasites, this low species richness found might be reflected in a low level of investment by the hosts in the basal immune defense, since investments in white blood cell production by mammals are influenced by the diversity of parasites in the environment. Additionally, considering that it might result in host vulnerability to other parasites that might be introduced through exotic or migratory host species, the monitoring of C. intermedia, including parasitological and immunological assessments, is recommended as a key component of conservation efforts.

  20. Le cobaye Cavia porcellus L., comme animal de boucherie au Cameroun

    Directory of Open Access Journals (Sweden)

    Ngou Ngoupayou, JD.

    1994-01-01

    Full Text Available Guinea Pig Cavia porcellus L. As A Meat Producing Animal In Cameroon. Guinea pig Cavia porcellus farming for meat production remains a marginalised activity in Cameroon in spite of the advantages this specie offers. With the view to promote its production, a national countrywide survey was carried out in order to evaluate the production systems and constraints. Traditional guinea pig farming appears to be a secondary household activity undertaken by small farmers basically women. The extensive production system which reveals no management practices, integrates very well in the agricultural systems (small livestock, food crops and natural forages production of the western highlands and southern forest zones of Cameroon. Guinea pig productivity remains low due to many constraints such as predation, uncontrolled breeding, inbreeding, poor feeding, negative selection and lack of veterinary care. Nevertheless, guinea pig farming plays an important role in the well being of low income village dwellers as food security, cash savings and socio-cultural values within the populations of South Cameroon. The promotion of its production requires an awareness of animal scientists, development authorities, as well as on station research if improved raising conditions.

  1. Fibroblasts Influence Survival and Therapeutic Response in a 3D Co-Culture Model.

    Science.gov (United States)

    Majety, Meher; Pradel, Leon P; Gies, Manuela; Ries, Carola H

    2015-01-01

    In recent years, evidence has indicated that the tumor microenvironment (TME) plays a significant role in tumor progression. Fibroblasts represent an abundant cell population in the TME and produce several growth factors and cytokines. Fibroblasts generate a suitable niche for tumor cell survival and metastasis under the influence of interactions between fibroblasts and tumor cells. Investigating these interactions requires suitable experimental systems to understand the cross-talk involved. Most in vitro experimental systems use 2D cell culture and trans-well assays to study these interactions even though these paradigms poorly represent the tumor, in which direct cell-cell contacts in 3D spaces naturally occur. Investigating these interactions in vivo is of limited value due to problems regarding the challenges caused by the species-specificity of many molecules. Thus, it is essential to use in vitro models in which human fibroblasts are co-cultured with tumor cells to understand their interactions. Here, we developed a 3D co-culture model that enables direct cell-cell contacts between pancreatic, breast and or lung tumor cells and human fibroblasts/ or tumor-associated fibroblasts (TAFs). We found that co-culturing with fibroblasts/TAFs increases the proliferation in of several types of cancer cells. We also observed that co-culture induces differential expression of soluble factors in a cancer type-specific manner. Treatment with blocking antibodies against selected factors or their receptors resulted in the inhibition of cancer cell proliferation in the co-cultures. Using our co-culture model, we further revealed that TAFs can influence the response to therapeutic agents in vitro. We suggest that this model can be reliably used as a tool to investigate the interactions between a tumor and the TME.

  2. Depot-dependent effects of adipose tissue explants on co-cultured hepatocytes.

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    Zhen-Yu Du

    Full Text Available We have developed an in vitro hepatocyte-adipose tissue explant (ATE co-culture model enabling examination of the effect of visceral and subcutaneous adipose tissues on primary rat hepatocytes. Initial analyses of inflammatory marker genes were performed in fractionated epididymal or inguinal adipose tissues. Expressions of inflammation related genes (IL-6, TNF-α, COX-2 were higher in the inguinal than the epididymal ATE. Similarly, expressions of marker genes of macrophage and monocyte (MPEG-1, CD68, F4/80, CD64 were higher in the stromal vascular fraction (SVF isolated from inguinal ATE than that from epididymal ATE. However, expressions of lipolysis related genes (ATGL, HSL, perilipin-1 were higher in the epididymal adipocytes than inguinal adipocytes. Moreover, secretion of IL-6 and PGE(2 was higher from inguinal ATEs than from epididymal ATEs. There was a trend that the total levels of IL-6, TNF-α and PGE(2 in the media from inguinal ATEs co-cultured with primary rat hepatocytes were higher than that in the media from epididymal ATEs co-cultured with hepatocytes, although the significant difference was only seen in PGE(2. Lipolysis, measured as glycerol release, was similar in the ATEs isolated from inguinal and epididymal adipose tissues when cultured alone, but the glycerol release was higher in the ATEs isolated from epididymal than from inguinal adipose tissue when co-cultured with hepatocytes. Compared to epididymal ATEs, the ATEs from inguinal adipose tissue elicited a stronger cytotoxic response and higher level of insulin resistance in the co-cultured hepatocytes. In conclusion, our results reveal depot-dependent effects of ATEs on co-cultured primary hepatocytes, which in part may be related to a more pronounced infiltration of stromal vascular cells (SVCs, particularly macrophages, in inguinal adipose tissue resulting in stronger responses in terms of hepatotoxicity and insulin-resistance.

  3. Response of a co-culture model of epithelial cells and gingival fibroblasts to zoledronic acid

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    Fernanda Gonçalves BASSO

    Full Text Available Abstract Osteonecrosis of the jaw is an adverse effect of bisphosphonates. While the etiopathogenesis of this condition has been investigated, the interactions and effects of bisphosphonates on oral mucosa cells remain unclear. It is hypothesized that cell culture models, such as co-culture or three-dimensional cell culture models, can provide valuable insight. Therefore, the aim of this study was to evaluate the effects of zoledronic acid (ZA on epithelial cells and gingival fibroblasts in a co-culture model. Briefly, epithelial cells were seeded on transwell inserts and gingival fibroblasts were seeded in the lower well of 24-well plates. The latter were treated with ZA (5 μM for 24 or 48 h. Cell viability and synthesis of the inflammatory chemokine, CCL2, were subsequently assessed. Data were subjected to statistical analysis with a 5% significance level. In the presence of ZA, the epithelial cells exhibited significant toxicity in both cell culture models and at both time points. However, greater cytotoxicity was observed in the co-culture model. Greater viability for the gingival fibroblasts was also associated with the co-culture model, and ZA-mediated toxicity was observed for the 48 h time point. ZA promoted a significant increase in CCL2 synthesis in both sets of cells, with greater CCL2 synthesis detected in the gingival fibroblasts. However, this effect was diminished in the co-culture model. Taken together, these results confirm the specific response patterns of the cells seeded in the co-culture model and also demonstrate the protective mechanism that is mediated by epithelial/mesenchymal cell interactions upon exposure to ZA.

  4. Response of a co-culture model of epithelial cells and gingival fibroblasts to zoledronic acid.

    Science.gov (United States)

    Basso, Fernanda Gonçalves; Soares, Diana Gabriela; Pansani, Taisa Nogueira; Turrioni, Ana Paula Silveira; Scheffel, Débora Lopes; Hebling, Josimeri; Costa, Carlos Alberto de Souza

    2016-11-28

    Osteonecrosis of the jaw is an adverse effect of bisphosphonates. While the etiopathogenesis of this condition has been investigated, the interactions and effects of bisphosphonates on oral mucosa cells remain unclear. It is hypothesized that cell culture models, such as co-culture or three-dimensional cell culture models, can provide valuable insight. Therefore, the aim of this study was to evaluate the effects of zoledronic acid (ZA) on epithelial cells and gingival fibroblasts in a co-culture model. Briefly, epithelial cells were seeded on transwell inserts and gingival fibroblasts were seeded in the lower well of 24-well plates. The latter were treated with ZA (5 μM) for 24 or 48 h. Cell viability and synthesis of the inflammatory chemokine, CCL2, were subsequently assessed. Data were subjected to statistical analysis with a 5% significance level. In the presence of ZA, the epithelial cells exhibited significant toxicity in both cell culture models and at both time points. However, greater cytotoxicity was observed in the co-culture model. Greater viability for the gingival fibroblasts was also associated with the co-culture model, and ZA-mediated toxicity was observed for the 48 h time point. ZA promoted a significant increase in CCL2 synthesis in both sets of cells, with greater CCL2 synthesis detected in the gingival fibroblasts. However, this effect was diminished in the co-culture model. Taken together, these results confirm the specific response patterns of the cells seeded in the co-culture model and also demonstrate the protective mechanism that is mediated by epithelial/mesenchymal cell interactions upon exposure to ZA.

  5. Label-free imaging to study phenotypic behavioural traits of cells in complex co-cultures

    Science.gov (United States)

    Suman, Rakesh; Smith, Gabrielle; Hazel, Kathryn E. A.; Kasprowicz, Richard; Coles, Mark; O'Toole, Peter; Chawla, Sangeeta

    2016-02-01

    Time-lapse imaging is a fundamental tool for studying cellular behaviours, however studies of primary cells in complex co-culture environments often requires fluorescent labelling and significant light exposure that can perturb their natural function over time. Here, we describe ptychographic phase imaging that permits prolonged label-free time-lapse imaging of microglia in the presence of neurons and astrocytes, which better resembles in vivo microenvironments. We demonstrate the use of ptychography as an assay to study the phenotypic behaviour of microglial cells in primary neuronal co-cultures through the addition of cyclosporine A, a potent immune-modulator.

  6. Co-culture of Gastric Organoids and Immortalized Stomach Mesenchymal Cells.

    Science.gov (United States)

    Bertaux-Skeirik, Nina; Centeno, Jomaris; Feng, Rui; Schumacher, Michael A; Shivdasani, Ramesh A; Zavros, Yana

    2016-01-01

    Three-dimensional primary epithelial-derived gastric organoids have recently been established as an important tool to study gastric development, physiology, and disease. Specifically, mouse-derived fundic gastric organoids (mFGOs) co-cultured with Immortalized Stomach Mesenchymal Cells (ISMCs) reflect expression patterns of mature fundic cell types seen in vivo, thus allowing for long-term in vitro studies of gastric epithelial cell physiology, regeneration, and bacterial-host interactions. Here, we describe the development and culture of mFGOs, co-cultured with ISMCs.

  7. Genética poblacional de cobayas de Colombia, Cavia spp. (Rodentia: Caviidae con marcadores moleculares RAPD

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    Héctor Aníbal Campos

    2008-09-01

    Full Text Available En el presente estudio, mostramos los primeros resultados moleculares de formas colombianas de Cavia. Claramente, la población silvestre de C. anolaimae fue genéticamente diferenciada de la forma doméstica, C. porcellus, tal como ha sido demostrado por otros autores utilizando resultados morfométricos, osteológicos y cariotípicos. Ambas especies mostraron un considerable nivel de diversidad genética, aunque el segundo taxon mostró niveles mayores de esta diversidad. Los niveles de heterogeneidad genética también fueron mayores entre las poblaciones de C. porcellus (F ST = 0.254 que entre las poblaciones de C. anolaimae (F ST = 0.118. Esos niveles significativos de heterogeneidad genética, y los consiguientes bajos niveles de flujo génico, fueron discutidos comparativamente con los resultados por otros autores analizando otros marcadores moleculares (citocromo-b mitocondrial. Los resultados aquí mostrados son coherentes con un complejo proceso de domesticación en Cavia porcellus.Population genetics of Colombian Guinea Pigs, Cavia spp. (Rodentia: Caviidae with RAPD molecular markers. The genus Cavia occurs in South America, mainly in grasslands.. We collected blood samples from 97 individuals in six field populations and analyzed them with RAPD molecular markers. One wild type (C. anolaimae was differentiated from the domestic form (C. porcellus, in agreement with other authors who used morphological, osteological and karyotipic results. Genetic diversity was considerable in both species, but higher in C. porcellus. The levels of genetic heterogeneity were also higher among the populations of C. porcellus (F ST = 0.254 than among the populations of C. anolaimae (F ST = 0.118. These significant levels of genetic heterogeneity, and the low levels of gene flow, were consistent with a complex domestication process for Cavia porcellus. Rev. Biol. Trop. 56 (3: 1481-1501. Epub 2008 September 30.

  8. Interlaboratorium vergelijking van het onderzoek naar Aeromonas spp. in drinkwater (tweede onderzoek)

    NARCIS (Netherlands)

    Versteegh JFM; During M; Heisterkamp SH; Havelaar AH

    1988-01-01

    Door middel van kwantitatief onderzoek naar Aeromonas-bacterien in drie verschillende monsters water (De Bilt, De Meent en De Laak) werd een vergelijking gemaakt van de resultaten van Aeromonas onderzoek in 15 laboratoria in het kader van het deelproject: "Aeromonas in drinkwater". Varian

  9. Aeromonas australiensis sp. nov., isolated from irrigation water.

    Science.gov (United States)

    Aravena-Román, Max; Beaz-Hidalgo, Roxana; Inglis, Timothy J J; Riley, Thomas V; Martínez-Murcia, Antonio J; Chang, Barbara J; Figueras, Maria Jose

    2013-06-01

    A Gram-negative, facultatively anaerobic bacillus, designated strain 266(T), was isolated from an irrigation water system in the south-west of Western Australia. Analysis of the 16S rRNA gene sequence confirmed that strain 266(T) belonged to the genus Aeromonas, with the nearest species being Aeromonas fluvialis (99.6% similarity to the type strain, with 6 nucleotide differences) followed by Aeromonas veronii and Aeromonas allosaccharophila (both 99.5%). Analysis of gyrB and rpoD sequences suggested that strain 266(T) formed a phylogenetic line independent of other species in the genus. This was confirmed using the concatenated sequences of six housekeeping genes (gyrB, rpoD, recA, dnaJ, gyrA and dnaX) that also indicated that A. veronii and A. allosaccharophila were the nearest relatives. DNA-DNA reassociation experiments and phenotypic analysis further supported the conclusion that strain 266(T) represents a novel species, for which the name Aeromonas australiensis sp. nov. is proposed, with type strain 266(T) (=CECT 8023(T) =LMG 26707(T)). [corrected].

  10. Enhancement of protective immunity in European eel(Anguilla anguilla)against Aeromonas hydrophila and Aeromonas sobria by a recombinant Aeromonas outer membrane protein

    Institute of Scientific and Technical Information of China (English)

    Ruizhang Guan; Jing Xiong; Wenshu Huang; Songlin Guo

    2011-01-01

    To develop a vaccine,which can simultaneously prevent the diseases caused by various pathogenic bacteria in fish,we try to find a conserved outer membrane protein(OMP)antigen from different bacterial pathogens.In this study,an OMP fragment of 747 by(named as Omp-G),whichwas highly conserved in seven Aeromonas OMP sequences from the NCBI database,was amplified by PCR from one Aeromonas sobria strain(1110)and two Aeromonas hydrophila strains(1327 and B33)with the designed specific primers.The sequence was cloned into pGEX-2T(6 × His-tag)vector,expressed in Escherichia coli system,and then the recombinant protein(named as rOmp-G)was purified with nickel chelating affinity chromatography.The purified romp-G showed a good immunogenicity in rabbits and well-conserved characteristics in these three pathogens by enzyme-linked immunosorbed assay.Furthermore,the rOmp-G also showed good immunogenicity in eels(Anguilla anguilla)for eliciting significantly increased specific antibodies(P<0.01),and providing higher protection efficiencies(P<0.05)after the pathogens challenge.The values of the relative percent survival in eels were 70% and 50% for two A.hydrophila strain challenge,and 75% for A.sobria strain challenge.This is the first report of a potential vaccination in eels that simultaneously provide protectiveness against different Aeromonas pathogens with a conserved partial OMP.

  11. Co-culture engineering for microbial biosynthesis of 3-amino-benzoic acid in Escherichia coli.

    Science.gov (United States)

    Zhang, Haoran; Stephanopoulos, Gregory

    2016-07-01

    3-amino-benzoic acid (3AB) is an important building block molecule for production of a wide range of important compounds such as natural products with various biological activities. In the present study, we established a microbial biosynthetic system for de novo 3AB production from the simple substrate glucose. First, the active 3AB biosynthetic pathway was reconstituted in the bacterium Escherichia coli, which resulted in the production of 1.5 mg/L 3AB. In an effort to improve the production, an E. coli-E. coli co-culture system was engineered to modularize the biosynthetic pathway between an upstream strain and an downstream strain. Specifically, the upstream biosynthetic module was contained in a fixed E. coli strain, whereas a series of E. coli strains were engineered to accommodate the downstream biosynthetic module and screened for optimal production performance. The best co-culture system was found to improve 3AB production by 15 fold, compared to the mono-culture approach. Further engineering of the co-culture system resulted in biosynthesis of 48 mg/L 3AB. Our results demonstrate co-culture engineering can be a powerful new approach in the broad field of metabolic engineering.

  12. Study of chondrogenic potential of stem cells in co-culture with chondrons

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    Parisa Nikpou

    2016-06-01

    Full Text Available Objective(s: Three-dimensional biomimetic scaffolds have widespread applications in biomedical tissue engineering due to similarity of their nanofibrous architecture to native extracellular matrix. Co-culture system has stimulatory effect on chondrogenesis of adult mesenchymal stem cells. This work presents a co-culture strategy using human articular chondrons and adipose-derived stem cells (ASCs from infrapatellar fat pad (IPFP for cartilage tissue production. Materials and Methods: Isolated stem cells were characterized by flowcytometry. Electrospun and polycaprolactone (PCL scaffolds (900 nm fiber diameter was obtained from Bon Yakhteh (Tehran- Iran and human infrapatellar fat pad-derived stem cells (IPFP-ASCs were seeded on them. IPFP- ASCs on scaffolds were co-cultured with articular chondrons using transwell. After 21 day, chondrogenic differentiation of stem cell was evaluated by determining the genes expression of collagen2, aggrecan and Indian hedgehog using real- time RT-PCR. Results: Genes expression of collagen2, aggrecan by IPFP-ASCs did not alter significantly in comparison with control group. Howevers, expression of Indian hedgehog decreased significantly compared to control group (P˂ 0.05. Conclusion: These findings indicate that chondrons obtained from osteoarthritic articular cartilage did not stimulate chondrogenic differentiation of IPFP-ASCs in co-culture.

  13. Boron nitride nanotube-mediated stimulation of cell co-culture on micro-engineered hydrogels.

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    Leonardo Ricotti

    Full Text Available In this paper, we describe the effects of the combination of topographical, mechanical, chemical and intracellular electrical stimuli on a co-culture of fibroblasts and skeletal muscle cells. The co-culture was anisotropically grown onto an engineered micro-grooved (10 µm-wide grooves polyacrylamide substrate, showing a precisely tuned Young's modulus (∼ 14 kPa and a small thickness (∼ 12 µm. We enhanced the co-culture properties through intracellular stimulation produced by piezoelectric nanostructures (i.e., boron nitride nanotubes activated by ultrasounds, thus exploiting the ability of boron nitride nanotubes to convert outer mechanical waves (such as ultrasounds in intracellular electrical stimuli, by exploiting the direct piezoelectric effect. We demonstrated that nanotubes were internalized by muscle cells and localized in both early and late endosomes, while they were not internalized by the underneath fibroblast layer. Muscle cell differentiation benefited from the synergic combination of topographical, mechanical, chemical and nanoparticle-based stimuli, showing good myotube development and alignment towards a preferential direction, as well as high expression of genes encoding key proteins for muscle contraction (i.e., actin and myosin. We also clarified the possible role of fibroblasts in this process, highlighting their response to the above mentioned physical stimuli in terms of gene expression and cytokine production. Finally, calcium imaging-based experiments demonstrated a higher functionality of the stimulated co-cultures.

  14. Co-culture systems-based strategies for articular cartilage tissue engineering.

    Science.gov (United States)

    Zhang, Yu; Liu, Shuyun; Guo, Weimin; Wang, Mingjie; Hao, Chunxiang; Gao, Shuang; Zhang, Xueliang; Li, Xu; Chen, Mingxue; Li, Penghao; Peng, Jiang; Lu, Shibi; Guo, Quanyi

    2017-05-26

    Cartilage engineering facilitates repair and regeneration of damaged cartilage using engineered tissue that restores the functional properties of the impaired joint. The seed cells used most frequently in tissue engineering, are chondrocytes and mesenchymal stem cells. Seed cells activity plays a key role in the regeneration of functional cartilage tissue. However, seed cells undergo undesirable changes after in vitro processing procedures, such as degeneration of cartilage cells and induced hypertrophy of mesenchymal stem cells, which hinder cartilage tissue engineering. Compared to monoculture, which does not mimic the in vivo cellular environment, co-culture technology provides a more realistic microenvironment in terms of various physical, chemical and biological factors. Co-culture technology is used in cartilage tissue engineering to overcome obstacles related to the degeneration of seed cells, and shows promise for cartilage regeneration and repair. In this review, we focus first on existing co-culture systems for cartilage tissue engineering and related fields, and discuss the conditions and mechanisms thereof. This is followed by methods for optimizing seed cell co-culture conditions to generate functional neo-cartilage tissue, which will lead to a new era in cartilage tissue engineering. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  15. Hydrogel microfluidic co-culture device for photothermal therapy and cancer migration.

    Science.gov (United States)

    Lee, Jong Min; Seo, Hye In; Bae, Jun Hyuk; Chung, Bong Geun

    2017-02-07

    We developed the photo-crosslinkable hydrogel microfluidic co-culture device to study photothermal therapy and cancer cell migration. To culture MCF7 human breast carcinoma cells and metastatic U87MG human glioblastoma in the microfluidic device, we used 10 w/v% gelatin methacrylate (GelMA) hydrogels as a semi-permeable physical barrier. We demonstrated the effect of gold nanorod on photothermal therapy of cancer cells in the microfluidic co-culture device. Interestingly, we observed that metastatic U87MG human glioblastoma largely migrated toward vascular endothelial growth factor (VEGF)-treated GelMA hydrogel-embedding microchannels. The main advantage of this hydrogel microfluidic co-culture device is to simultaneously analyze the physiological migration behaviors of two cancer cells with different physiochemical motilities and study gold nanorod-mediated photothermal therapy effect. Therefore, this hydrogel microfluidic co-culture device could be a potentially powerful tool for photothermal therapy and cancer cell migration applications. This article is protected by copyright. All rights reserved.

  16. Comparative proteomic analysis of experimental evolution of the Bacillus cereus-Ketogulonicigenium vulgare co-culture.

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    Qian Ma

    Full Text Available The microbial co-culture system composing of Ketogulonicigenium vulgare and Bacillus cereus was widely adopted in industry for the production of 2-keto-gulonic acid (2-KGA, the precursor of vitamin C. We found serial subcultivation of the co-culture could enhance the yield of 2-KGA by 16% in comparison to that of the ancestral co-culture. To elucidate the evolutionary dynamics and interaction mechanisms of the two microbes, we performed iTRAQ-based quantitative proteomic analyses of the pure cultures of K. vulgare, B. cereus and their co-culture during serial subcultivation. Hierarchy cluster analyses of the proteomic data showed that the expression level of a number of crucial proteins associated with sorbose conversion and oligopeptide transport was significantly enhanced by the experimental evolution. In particular, the expression level of sorbose/sorbosone dehydrogenase was enhanced in the evolved K. vulgare, while the expression level of InhA and the transport efficiency of oligopeptides were increased in the evolved B. cereus. The decreased sporulating protein expression and increased peptide transporter expression observed in evolved B. cereus, together with the increased amino acids synthesis in evolved K. vulgare suggested that serial subcultivation result in enhanced synergistic cooperation between K. vulgare and B. cereus, enabling an increased production of 2-KGA.

  17. Evaluation of Medicinal Plant Hepatotoxicity in Co-cultures of Hepatocytes and Monocytes

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    Bashar Saad

    2006-01-01

    Full Text Available Non-parenchymal cells might play an important role in the modulation of xenobiotic metabolism in liver and its pharmacological and toxicological consequences. Therefore, the role of cell-to-cell interactions in herbal induced liver toxicity was investigated in monocultures of cells from the human hepatocyte cell line (HepG2 and in co-cultures of cells from the HepG2 cell line and cells from the human monocyte cell line (THP1. Cells were treated with various concentrations (1–500 µg ml−1 of extracts of Pistacia palaestina, Juglans regia and Quercus ithaburensis for 24 h. Extracts from Cleome droserifolia, a known toxic plant, were taken as positive control. In the co-culture system, toxic effects were observed after exposure to extracts of Pistacia palestina and C. droserifolia. These two extracts significantly reduced by cell viability as measured the MTT test and the LDH assay. Whereas in hepatocyte cultures, only extracts of C. droserifolia were found to affect the cell viability. The production levels of albumin from hepatocytes were not affected by treatment with plant extracts in both culture systems. It seems that the observed reduction in cell viability after exposure to extracts of P. palestina in co-cultures but not in monocultures is a result of monocyte-derived factors. The use of liver cell co-cultures is therefore a useful approach to investigate the influence of intercellular communication on xenobiotic metabolism in liver.

  18. Effects of Tibolone Metabolites on Human Endometrial Cell Lines in Co-Culture

    Science.gov (United States)

    Barbier, Claire; Kloosterboer, Helenius J.; Kaufman, David G.

    2010-01-01

    In human endometrium, cell proliferation is regulated by ovarian steroids through heterotypic interactions between stromal and epithelial cells populating this tissue. We tested the proliferative effects of tibolone and its metabolites using endometrial co-cultures that mimic the normal proliferative response to hormones. We found that both the Δ4-tibolone metabolite and the pure progestin ORG2058 counteract estradiol-driven epithelial cell proliferation. Surprisingly, the estrogen receptor binding 3-hydroxyl-metabolites of tibolone also counteracted estradiol-driven proliferation. Inhibition of proliferation by 3β-OH-tibolone was abrogated by low doses of the progesterone receptor antagonist mifepristone, This suggests that 3β-OH-tibolone is converted to a progestagenic metabolite. We found that the stromal cells used in the co-cultures express high levels of the ketosteroid dehydrogenase, AKR1C2, which is able to oxidize 3β-OH-tibolone back to tibolone. Thus the unexpected progestagenic effect of 3β-OH-tibolone in these co-cultures may be due to metabolic activity present in the stromal cells of the co-cultures. PMID:18212357

  19. Boosting dark fermentation with co-cultures of extreme thermophiles for biohythane production from garden waste.

    Science.gov (United States)

    Abreu, Angela A; Tavares, Fábio; Alves, Maria Madalena; Pereira, Maria Alcina

    2016-11-01

    Proof of principle of biohythane and potential energy production from garden waste (GW) is demonstrated in this study in a two-step process coupling dark fermentation and anaerobic digestion. The synergistic effect of using co-cultures of extreme thermophiles to intensify biohydrogen dark fermentation is demonstrated using xylose, cellobiose and GW. Co-culture of Caldicellulosiruptor saccharolyticus and Thermotoga maritima showed higher hydrogen production yields from xylose (2.7±0.1molmol(-1) total sugar) and cellobiose (4.8±0.3molmol(-1) total sugar) compared to individual cultures. Co-culture of extreme thermophiles C. saccharolyticus and Caldicellulosiruptor bescii increased synergistically the hydrogen production yield from GW (98.3±6.9Lkg(-1) (VS)) compared to individual cultures and co-culture of T. maritima and C. saccharolyticus. The biochemical methane potential of the fermentation end-products was 322±10Lkg(-1) (CODt). Biohythane, a biogas enriched with 15% hydrogen could be obtained from GW, yielding a potential energy generation of 22.2MJkg(-1) (VS).

  20. Functional 3-D cardiac co-culture model using bioactive chitosan nanofiber scaffolds.

    Science.gov (United States)

    Hussain, Ali; Collins, George; Yip, Derek; Cho, Cheul H

    2013-02-01

    The in vitro generation of a three-dimensional (3-D) myocardial tissue-like construct employing cells, biomaterials, and biomolecules is a promising strategy in cardiac tissue regeneration, drug testing, and tissue engineering applications. Despite significant progress in this field, current cardiac tissue models are not yet able to stably maintain functional characteristics of cardiomyocytes for long-term culture and therapeutic purposes. The objective of this study was to fabricate bioactive 3-D chitosan nanofiber scaffolds using an electrospinning technique and exploring its potential for long-term cardiac function in the 3-D co-culture model. Chitosan is a natural polysaccharide biomaterial that is biocompatible, biodegradable, non-toxic, and cost effective. Electrospun chitosan was utilized to provide structural scaffolding characterized by scale and architectural resemblance to the extracellular matrix (ECM) in vivo. The chitosan fibers were coated with fibronectin via adsorption in order to enhance cellular adhesion to the fibers and migration into the interfibrous milieu. Ventricular cardiomyocytes were harvested from neonatal rats and studied in various culture conditions (i.e., mono- and co-cultures) for their viability and function. Cellular morphology and functionality were examined using immunofluorescent staining for alpha-sarcomeric actin (SM-actin) and gap junction protein, Connexin-43 (Cx43). Scanning electron microscopy (SEM) and light microscopy were used to investigate cellular morphology, spatial organization, and contractions. Calcium indicator was used to monitor calcium ion flux of beating cardiomyocytes. The results demonstrate that the chitosan nanofibers retained their cylindrical morphology in long-term cell cultures and exhibited good cellular attachment and spreading in the presence of adhesion molecule, fibronectin. Cardiomyocyte mono-cultures resulted in loss of cardiomyocyte polarity and islands of non-coherent contractions. However

  1. Co-culture of 3D tumor spheroids with fibroblasts as a model for epithelial–mesenchymal transition in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sun-Ah, E-mail: j.sarah.k@gmail.com [Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Lee, Eun Kyung, E-mail: leeek@catholic.ac.kr [Department of Biochemistry, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Cancer Evolution Research Center, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Kuh, Hyo-Jeong, E-mail: hkuh@catholic.ac.kr [Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Cancer Evolution Research Center, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of)

    2015-07-15

    Epithelial–mesenchymal transition (EMT) acts as a facilitator of metastatic dissemination in the invasive margin of malignant tumors where active tumor–stromal crosstalks take place. Co-cultures of cancer cells with cancer-associated fibroblasts (CAFs) are often used as in vitro models of EMT. We established a tumor–fibroblast proximity co-culture using HT-29 tumor spheroids (TSs) with CCD-18co fibroblasts. When co-cultured with TSs, CCD-18co appeared activated, and proliferative activity as well as cell migration increased. Expression of fibronectin increased whereas laminin and type I collagen decreased in TSs co-cultured with fibroblasts compared to TSs alone, closely resembling the margin of in vivo xenograft tissue. Active TGFβ1 in culture media significantly increased in TS co-cultures but not in 2D co-cultures of cancer cells–fibroblasts, indicating that 3D context-associated factors from TSs may be crucial to crosstalks between cancer cells and fibroblasts. We also observed in TSs co-cultured with fibroblasts increased expression of α-SMA, EGFR and CTGF; reduced expression of membranous β-catenin and E-cadherin, together suggesting an EMT-like changes similar to a marginal region of xenograft tissue in vivo. Overall, our in vitro TS–fibroblast proximity co-culture mimics the EMT-state of the invasive margin of in vivo tumors in early metastasis. - Highlights: • An adjacent co-culture of tumor spheroids and fibroblasts is presented as EMT model. • Activation of fibroblasts and increased cell migration were shown in co-culture. • Expression of EMT-related factors in co-culture was similar to that in tumor tissue. • Crosstalk between spheroids and fibroblasts was demonstrated by secretome analysis.

  2. Identificação e resistência a antimicrobianos de espécies de Aeromonas móveis isoladas de peixes e ambientes aquáticos Identification and antimicrobial resistance of motile Aeromonas isolated from fish and aquatic environment

    Directory of Open Access Journals (Sweden)

    Daniela Hirsch

    2006-12-01

    Full Text Available Com o objetivo de se verificar a diversidade de espécies de Aeromonas móveis e seu perfil de susceptibilidade a antimicrobianos em pisciculturas comerciais, foram selecionadas oito tilapiculturas localizadas na região do Alto Rio Grande, Minas Gerais. De cada propriedade foram coletadas três amostras de peixes em estádio de pré-abate (vivos e saudáveis, uma amostra de água do tanque e uma amostra da água de abastecimento do sistema. De cada peixe foram coletadas amostras de lavado superficial e do parênquima renal. Diluições seriadas adequadas de cada amostra foram plaqueadas em TSA-ampicilina (10 mg/l e as amostras de rim em Ágar Sangue de cavalo a 5%. A partir de colônias isoladas positivas para o teste da oxidase foram realizados testes para identificação do gênero (testes presuntivos e das espécies de Aeromonas (testes bioquímicos. O perfil de antibiograma foi obtido pelo teste de difusão de discos de antibióticos em Ágar Mueller Hinton. Foram obtidos 75 isolados diferenciados em nove espécies de Aeromonas: A. jandaei, A. hydrophila, A. trota, A. caviae, A. sobria, A. eucrenophila, A. veronii bt veronii, A. schubertii, A. media, além de amostras classificadas como Aeromonas atípicas. Do total isolado, oito amostras foram provenientes da superfície corpórea de peixes, 14 da água de abastecimento e 53 da água do tanque. Não houve isolamento a partir dos espécimes de parênquima renal. Em relação ao perfil de resistência, 93% dos isolados foram resistentes à eritromicina, 36% à tetraciclina, 13% ao ác. nalidíxico, 9% à gentamicina, 8% à nitrofurantoína, 8% à canamicina, 5% à norfloxacina, 4% ao cloranfenicol e 3% às sulfonamidas. Dentre os isolados analisados, 43% apresentaram índice de múltipla resistência a antimicrobianos (MAR igual ou superior a 22%, ou seja, resistência a dois ou mais drogas das nove testadas. Os dados apontam para um risco iminente, tanto pelo isolamento de amostras

  3. Co-cultured production of lignin-modifying enzymes with white-rot fungi.

    Science.gov (United States)

    Qi-He, Chen; Krügener, Sven; Hirth, Thomas; Rupp, Steffen; Zibek, Susanne

    2011-09-01

    Co-cultivation was a potential strategy in lignocellulolytic biodegradation with producing high activity enzymes due to their synergistic action. The objective of this study was to investigate the rarely understood effects of co-culturing of two white-rot fungi on lignin-modifying enzymes (LMEs) production. Six species, Bjerkandera adusta, Phlebia radiata, Pleurotus ostreatus, Dichomitus squalens, Hypoxylon fragiforme and Pleurotus eryngii, were cultured in pairs to study the production of LMEs. The paired hyphal interaction observed showed that P. eryngii is not suitable for co-growth. The use of agar plates containing dye RBBR showed elevated decolourisation at the confrontation zone between mycelia. Laccase was significantly stimulated only in the co-culture of P. radiata with D. squalens under submerged cultivation; the highest value was measured after 4 days of incubation (120 U mg(-1)). The improved productions of MnP and LiP were simultaneously observed at the co-culture of P. ostreatus and P. radiata (MnP = 800 nkat L(-1) after 4 days of incubation; LiP = 60 nkat L(-1) after 7 days of incubation), though it was not a good producer of laccase. P. ostreatus appeared to possess specific potential to be used in co-cultured production of LMEs. The phenotype of LMEs production was not only dependent on the species used but also regulated by different nutritions available in the culture medium. The present data will provide evidence for illustrating the regulatory roles of C/N on LMEs production under the co-cultures' circumstances.

  4. Co-culture with Sertoli cells promotes proliferation and migration of umbilical cord mesenchymal stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Fenxi, E-mail: fxzhang0824@gmail.com [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Hong, Yan; Liang, Wenmei [Department of Histology and Embryology, Guiyang Medical University, Guizhou 550004, People' s Republic of China (China); Ren, Tongming [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Jing, Suhua [ICU Center, The Third Hospital of Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Lin, Juntang [Stem Cell Center, Xinxiang Medical University, Henan 453003, People' s Republic of China (China)

    2012-10-12

    Highlights: Black-Right-Pointing-Pointer Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). Black-Right-Pointing-Pointer Presence of SCs dramatically increased proliferation and migration of UCMSCs. Black-Right-Pointing-Pointer Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of 'nurse' cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.

  5. Co-culture: A quick approach for isolation of street rabies virus in murine neuroblastoma cells

    Directory of Open Access Journals (Sweden)

    A. Sasikalaveni

    2015-05-01

    Full Text Available Background: Laboratory detection of rabies in most cases is based on detection of the antigen by fluorescent antibody test, however, in weak positive cases confirmative laboratory diagnosis depends on widely accepted mouse inoculation test. Cell lines like neuroblastoma have been used to isolate the virus with greater success not only to target for diagnosis, but also for molecular studies that determine the epidemiology of the circulating street rabies strains and in studies that look at the efficiency of the developed monoclonal antibodies to neutralize the different rabies strains. Due to the recent issues in obtaining ethical permission for mouse experimentation, and also the passages required in the cell lines to isolate the virus, we report herewith a co-culture protocol using the murine neuroblastoma (MNA cells, which enable quicker isolation of street rabies virus with minimum passages. Objective: This study is not to have an alternative diagnostic assay, but an approach to produce sufficient amount of rabies virus in minimum passages by a co-culture approach in MNA cells. Materials and Methods: The MNA cells are co-cultured by topping the normal cells with infected cells every 48 h and the infectivity was followed up by performing direct fluorescent-antibody test. Results: The co-culture approach results in 100% infectivity and hence the use of live mouse for experimentation could be avoided. Conclusion: Co-culture method provides an alternative for the situations with limited sample volume and for the quicker isolation of virus which warrants the wild type strains without much modification.

  6. Dynamic three-dimensional micropatterned cell co-cultures within photocurable and chemically degradable hydrogels.

    Science.gov (United States)

    Sugiura, Shinji; Cha, Jae Min; Yanagawa, Fumiki; Zorlutuna, Pinar; Bae, Hojae; Khademhosseini, Ali

    2016-08-01

    In this paper we report on the development of dynamically controlled three-dimensional (3D) micropatterned cellular co-cultures within photocurable and chemically degradable hydrogels. Specifically, we generated dynamic co-cultures of micropatterned murine embryonic stem (mES) cells with human hepatocellular carcinoma (HepG2) cells within 3D hydrogels. HepG2 cells were used due to their ability to direct the differentiation of mES cells through secreted paracrine factors. To generate dynamic co-cultures, mES cells were first encapsulated within micropatterned photocurable poly(ethylene glycol) (PEG) hydrogels. These micropatterned cell-laden PEG hydrogels were subsequently surrounded by calcium alginate (Ca-Alg) hydrogels containing HepG2 cells. After 4 days, the co-culture step was halted by exposing the system to sodium citrate solution, which removed the alginate gels and the encapsulated HepG2 cells. The encapsulated mES cells were then maintained in the resulting cultures for 16 days and cardiac differentiation was analysed. We observed that the mES cells that were exposed to HepG2 cells in the co-cultures generated cells with higher expression of cardiac genes and proteins, as well as increased spontaneous beating. Due to its ability to control the 3D microenvironment of cells in a spatially and temporally regulated manner, the method presented in this study is useful for a range of cell-culture applications related to tissue engineering and regenerative medicine. Copyright © 2013 John Wiley & Sons, Ltd.

  7. The impact of quorum sensing on the virulence of Aeromonas hydrophila and Aeromonas salmonicida towards burbot (Lota lota L.) larvae.

    Science.gov (United States)

    Natrah, F M I; Alam, Md Iftakharul; Pawar, Sushant; Harzevili, A Shiri; Nevejan, Nancy; Boon, Nico; Sorgeloos, Patrick; Bossier, Peter; Defoirdt, Tom

    2012-09-14

    In this study, the link between quorum sensing in Aeromonas spp. and its virulence towards burbot (Lota lota) was investigated. High mortality occurred in burbot juveniles challenged with Aeromonas salmonicida HN-00, but not in juveniles challenged with Aeromonas hydrophila AH-1N. Meanwhile, both A. hydrophila AH-1N and A. salmonicida HN-00 were virulent towards larvae. The effect of quorum sensing on the virulence of A. hydrophila AH-1N towards burbot larvae was further investigated using quorum sensing mutants (N-(butyryl)-L-homoserine lactone production and receptor mutants). Challenge with these mutants resulted in higher survival of burbot larvae when compared to challenge with the wild type, and the addition of the signal molecule N-butyryl-L-homoserine lactone restored the virulence of the quorum sensing production mutant. Moreover, quorum sensing inhibitors protected the burbot larvae from both Aeromonas strains. Finally, the freshwater micro-algae Chlorella saccharophila and Chlamydomonas reinhardtii, which are able to interfere with quorum sensing, also protected burbot from the pathogens. However, QS interference was unlikely to be the only mechanism. This study revealed that the virulence of Aeromonas spp. towards burbot is regulated by quorum sensing and that quorum sensing inhibitors and micro-algae are promising biocontrol agents.

  8. Reclassification of Aeromonas hydrophila subsp. dhakensis Huys et al. 2002 and Aeromonas aquariorum Martínez-Murcia et al. 2008 as Aeromonas dhakensis sp. nov. comb nov. and emendation of the species Aeromonas hydrophila.

    Science.gov (United States)

    Beaz-Hidalgo, Roxana; Martínez-Murcia, Antonio; Figueras, Maria José

    2013-05-01

    Previous studies indicate that Aeromonas aquariorum and Aeromonas hydrophila subsp. dhakensis are the same taxon and suggest that they should be synonymized. Using a polyphasic approach, the phenotypic and phylogenetic relationship of A. aquariorum with the 3 defined A. hydrophila subspecies (i.e. dhakensis, hydrophila, ranae) was investigated. Phylogenetic trees derived from the 16S rRNA, rpoD or gyrB genes and a multilocus phylogenetic analysis (with the concatenated sequences of gyrB, rpoD, recA, dnaJ and gyrA) confirmed that both A. aquariorum and A. hydrophila subsp. dhakensis are a unique taxon, different from the other A. hydrophila subspecies, corroborating the phenotypic and DNA-DNA hybridization (DDH) results. A formal synonymization of A. aquariorum and A. hydrophila subsp. dhakensis and a reclassification of both as Aeromonas dhakensis sp. nov. comb nov. is therefore proposed.

  9. Susceptibility of Aeromonas Hydophila Isolates to Antimicrobial Drugs

    Directory of Open Access Journals (Sweden)

    Igor Stojanov

    2010-05-01

    Full Text Available Aeromonas hydrophila is a microorganism widely distributed in nature: in water, soil, food. It is also part of the normal bacterial flora of many animals. As an opportune microorganism it is a secondary biological agent that contributes to the occurrence of a fish disease and its deterioration. Frequently, its presence is an indication of bad zoohygiene and zootechnical conditions in fish ponds. Reduced quality and quantity of feed, mechanical injuries, parasitosis, seasonal oscillation in temperature present some of the factors that produce favorable conditions for bacterial proliferation of aeromonas in fish ponds, so clinical symptoms of the disease occur. Aeromonas is almost always present in clinical isolates and may be unjustly accused for bad health of fish. Antibiotic therapy is applied even when the clinical findings are clear, what certainly effects the susceptibility to chemotherapeutics. The subject of our work was bacteriological examination of the material obtained from the carps with the observed skin changes and the carps without these changes. Also, antimicrobial susceptibility of Aeromonas hydrophila was tested. The aim of this research was to determined the presence of Aeromonas hydrophilia in the carp ponds and to test antibiotic susceptibility. The material consisted of the samples from the fish ponds where the carps were with and without changed skin. The method the isolation of Aeromonas hydrophila was used. The diffusion disk technique was used for testing antibiotic susceptibility. The isolates were tested for their susceptibility to Florephenikol, Flumequine, Olaqindox and Oxitetracycline. The obtained results point that antimicrobial susceptibility was the same regardless of the origin of the samples, i.e. the resistance was the same for both groups of samples (the strains isolated from the fish with skin changes and the strains from fish without changes on skin. The strains were highly resistant: 35% were resistant to

  10. A defined co-culture of Geobacter sulfurreducens and Escherichia coli in a membrane-less microbial fuel cell.

    Science.gov (United States)

    Bourdakos, Nicholas; Marsili, Enrico; Mahadevan, Radhakrishnan

    2014-04-01

    Wastewater-fed microbial fuel cells (MFCs) are a promising technology to treat low-organic carbon wastewater and recover part of the chemical energy in wastewater as electrical power. However, the interactions between electrochemically active and fermentative microorganisms cannot be easily studied in wastewater-fed MFCs because of their complex microbial communities. Defined co-culture MFCs provide a detailed understanding of such interactions. In this study, we characterize the extracellular metabolites in laboratory-scale membrane-less MFCs inoculated with Geobacter sulfurreducens and Escherichia coli co-culture and compare them with pure culture MFCs. G. sulfurreducens MFCs are sparged to maintain anaerobic conditions, while co-culture MFCs rely on E. coli for oxygen removal. G. sulfurreducens MFCs have a power output of 128 mW m(-2) , compared to 63 mW m(-2) from the co-culture MFCs. Analysis of metabolites shows that succinate production in co-culture MFCs decreases current production by G. sulfurreducens and that the removal of succinate is responsible for the increased current density in the late co-culture MFCs. Interestingly, pH adjustment is not required for co-culture MFCs but a base addition is necessary for E. coli MFCs and cultures in vials. Our results show that defined co-culture MFCs provide clear insights into metabolic interactions among bacteria while maintaining a low operational complexity.

  11. Development of melanocye-keratinocyte co-culture model for controls and vitiligo to assess regulators of pigmentation and melanocytes

    Directory of Open Access Journals (Sweden)

    Ravinder Kumar

    2012-01-01

    Full Text Available Background: There is a need to develop an in vitro skin models which can be used as alternative system for research and testing pharmacological products in place of laboratory animals. Therefore to study the biology and pathophysiology of pigmentation and vitiligo, reliable in vitro skin pigmentation models are required. Aim: In this study, we used primary cultured melanocytes and keratinocytes to prepare the skin co-culture model in control and vitiligo patients. Methods: The skin grafts were taken from control and patients of vitiligo. In vitro co-culture was prepared after culturing primary melanocytes and keratinocytes. Co- cultures were treated with melanogenic stimulators and inhibitors and after that tyrosinase assay, MTT assay and melanin content assay were performed. Results: Melanocytes and keratinocytes were successfully cultured from control and vitiligo patients and after that co-culture models were prepared. After treatment of co-culture model with melanogenic stimulator we found that tyrosinase activity, cell proliferation and melanin content increased whereas after treatment with melanogenic inhibitor, tyrosinase activity, cell proliferation and melanin content decreased. We also found some differences in the control co-culture model and vitiligo co-culture model. Conclusion: We successfully constructed in vitro co-culture pigmentation model for control and vitiligo patients using primary cultured melanocytes and keratinocytes. The use of primary melanocytes and keratinocytes is more appropriate over the use of transformed cells. The only limitation of these models is that these can be used for screening small numbers of compounds.

  12. Influence of co-culture on osteogenesis and angiogenesis of bone marrow mesenchymal stem cells and aortic endothelial cells.

    Science.gov (United States)

    Gurel Pekozer, Gorke; Torun Kose, Gamze; Hasirci, Vasif

    2016-11-01

    Co-culture of bone forming cells and endothelial cells to induce pre-vascularization is one of the strategies used to solve the insufficient vascularization problem in bone tissue engineering attempts. In the study, primary cells isolated from 2 different tissues of the same animal, rat bone marrow stem cells (RBMSCs) and rat aortic endothelial cells (RAECs) were co-cultured to study the effects of co-culturing on both osteogenesis and angiogenesis. The formation of tube like structure in 2D culture was observed for the first time in the literature by the co-culture of primary cells from the same animal and also osteogenesis and angiogenesis were investigated at the same time by using this co-culture system. Co-cultured cells mineralized and formed microvasculature beginning from 14days of incubation. After 28days of incubation in the osteogenic medium, expression of osteogenic genes in co-cultures was significantly upregulated compared to RBMSCs cultured alone. These results suggest that the co-culture of endothelial cells with mesenchymal stem cells induces both osteogenesis and angiogenesis.

  13. 基于CAViaR模型的银行间质押回购利率风险研究

    Institute of Scientific and Technical Information of China (English)

    张俊; 王晓莹

    2016-01-01

    Value at Risk is widely used in commercial bank interest rate risk management, but domestic commercial banks are still confined to use parameter method (the family of GARCH) and the historical simulation method to calculate the Value at Risk at present. Based on the overnight repo rate as the research object, this paper study the pledged repo operation law and wave characteristics of interest rate risk by constructing CAViaR risk measurement model. The results found that, CAViaR model can better depict the risk prediction effect of pledged repo rate risk. Based on the back test results found that, AS model is the optimal model in estimating China pledged repo rate risk. In addition, based on the overnight repo rate from 2006 to 2014 in VaR value analysis, this paper confirms the CAViaR model can better fitting of monetary market rate changes and the size of interest rate risk.%“在险价值”管理方法在商业银行利率风险管理中被广泛应用,但目前国内商业银行在计算“在险价值”时仍局限于参数方法(GARCH族模型)和历史模拟法。以隔夜质押回购利率作为研究对象,构建基于CAViaR的质押回购利率风险计量模型,研究质押回购利率风险的运行规律和波动特征。研究结果发现,CAViaR模型的风险预测效果能够较好地刻画质押回购利率的利率风险。基于后测检验结果发现,AS模型在估计我国质押回购利率风险时表现最优。此外,基于2006—2014年隔夜回购利率VaR值变化趋势的分析,证实了CAViaR模型能够较好地拟合货币市场利率变化情况及其利率风险的大小。

  14. Characterization of fetal growth by repeated ultrasound measurements in the wild guinea pig (Cavia aperea).

    Science.gov (United States)

    Schumann, K; Guenther, A; Göritz, F; Jewgenow, K

    2014-08-01

    Fetal growth during pregnancy has previously been studied in the domesticated guinea pig (Cavia aperea f. porcellus) after dissecting pregnant females, but there are no studies describing the fetal growth in their wild progenitor, the wild guinea pig (C aperea). In this study, 50 pregnancies of wild guinea pig sows were investigated using modern ultrasound technique. The two most common fetal growth parameters (biparietal diameter [BPD] and crown-rump-length [CRL]) and uterine position were measured. Data revealed similar fetal growth patterns in the wild guinea pig and domesticated guinea pig in the investigated gestation period, although they differ in reproductive milestones such as gestation length (average duration of pregnancy 68 days), average birth weight, and litter mass. In this study, pregnancy lasted on average 60.2 days with a variance of less than a day (0.96 days). The measured fetal growth parameters are strongly correlated with each (R = 0.91; P guinea pig.

  15. A new karyotype for the genus Cavia from a southern island of Brazil (Rodentia - Caviidae

    Directory of Open Access Journals (Sweden)

    A. Gava

    1998-03-01

    Full Text Available Intraspecific karyotype variation in mammal species is very common and often caused by centromeric fusion of acrocentric chromosomes. We describe here a new karyotype 2n = 62 (FN = 112 for the genus Cavia from the Moleques do Sul Islands, of the southern coast of Brazil. We analyzed two male and four female karyotypes that had twenty-four biarmed pairs and six pairs of acrocentric chromosomes. The sexual pair consisted of a metacentric X-chromosome and a large acrocentric Y. C-bands were found in the centromeric and pericentromeric regions of almost all chromosomes, except for some small biarmed and acrocentric ones. Nucleolus organizer regions appeared in two biarmed chromosomes, and G-banding patterns were also seen.RESUMO A variação cariotípica nas espécies de mamíferos é bastante comum e geralmente causada pela fusão de cromossomos acrocêntricos. Foi descrito neste trabalho um novo cariótipo, com 2n = 62 e FN = 112, para o gênero Cavia proveniente das ilhas Moleques do Sul, da costa sul do Brasil. Foram analisados os cariótipos de dois machos e quatro fêmeas que possuiam 24 pares de cromossomos com dois braços e seis pares de acrocêntricos. O par sexual era constituído por um cromossomo X metacêntrico grande e um Y acrocêntrico. As bandas C estavam localizadas nas regiões centroméricas e pericentroméricas da maioria dos cromossomos, com exceção de alguns acrocêntricos e os cromossomos de dois braços menores. As regiões organizadoras de nucléolo ocorreram em dois cromossomos com dois braços e o padrão de bandamento G foi também apresentado.

  16. Phosphoproteomic analysis of the Chlamydia caviae elementary body and reticulate body forms.

    Science.gov (United States)

    Fisher, Derek J; Adams, Nancy E; Maurelli, Anthony T

    2015-08-01

    Chlamydia are Gram-negative, obligate intracellular bacteria responsible for significant diseases in humans and economically important domestic animals. These pathogens undergo a unique biphasic developmental cycle transitioning between the environmentally stable elementary body (EB) and the replicative intracellular reticulate body (RB), a conversion that appears to require extensive regulation of protein synthesis and function. However, Chlamydia possess a limited number of canonical mechanisms of transcriptional regulation. Ser/Thr/Tyr phosphorylation of proteins in bacteria has been increasingly recognized as an important mechanism of post-translational control of protein function. We utilized 2D gel electrophoresis coupled with phosphoprotein staining and MALDI-TOF/TOF analysis to map the phosphoproteome of the EB and RB forms of Chlamydia caviae. Forty-two non-redundant phosphorylated proteins were identified (some proteins were present in multiple locations within the gels). Thirty-four phosphorylated proteins were identified in EBs, including proteins found in central metabolism and protein synthesis, Chlamydia-specific hypothetical proteins and virulence-related proteins. Eleven phosphorylated proteins were identified in RBs, mostly involved in protein synthesis and folding and a single virulence-related protein. Only three phosphoproteins were found in both EB and RB phosphoproteomes. Collectively, 41 of 42 C. caviae phosphoproteins were present across Chlamydia species, consistent with the existence of a conserved chlamydial phosphoproteome. The abundance of stage-specific phosphoproteins suggests that protein phosphorylation may play a role in regulating the function of developmental-stage-specific proteins and/or may function in concert with other factors in directing EB-RB transitions.

  17. Caracterização de Aeromonas spp isoladas de neonatos hospitalizados Characterization of Aeromonas spp isolates from newborns hospitalized

    OpenAIRE

    2008-01-01

    Aeromonas spp é reconhecida como patogênica para o homem após o consumo de água e alimentos contaminados. Na presente investigação, foram avaliadas 2.323 amostras de swabs retais de neonatos hospitalizados no Rio de Janeiro objetivando o isolamento de Aeromonas. As amostras foram coletadas e enviadas ao Laboratório de Referência Nacional de Cólera e outras enteroinfecções bacterianas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz. Os swabs foram submetidos ao enriquecimento em água peptonada ...

  18. Diarrea del viajero asociada a Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    M. Martín Delgado

    2001-06-01

    Full Text Available Aeromonas hydrophila es una bacteria que se encuentra frecuentemente en aguas dulces y salobres. Desde hace varios años se conoce que determinadas cepas de la especie pueden causar enfermedad en pescados y en anfibios, existiendo cierta controversia sobre su patogeneicidad en humanos. No obstante, puede dar lugar a infección en personas, que la adquieren bien a través de heridas abiertas o bien por ingestión de alimentos o agua contaminados. Sin embargo, en prácticamente la totalidad de las publicaciones se hace referencia a casos aislados más que a brotes producidos por esta bacteria. Generalmente la infección por A. hydrophila da lugar a gastroenteritis en personas sanas, pudiendo generar una septicemia en individuos inmunodeprimidos.En 1998 se produjo una alarma en el sector hotelero por un incremento considerable de los casos de gastroenteritis entre los turistas que acudían durante sus vacaciones a determinadas zonas costeras de la isla de Tenerife.Tras tener conocimiento de la situación, la Dirección General de Salud Pública inició una investigación epidemiológica con el objetivo de conocer la magnitud real del problema detectado e identificar los factores asociados a la misma, para poner en marcha las medidas correctoras y de control necesarias. El análisis inicial permitió determinar que se trataba de un proceso de gran magnitud en su extensión aunque de carácter leve, que afectaba únicamente a los turistas en una zona costera en concreto, compatible por lo tanto con la diarrea del viajero, planteándose un origen probablemente hídrico.

  19. Zooarqueología y tafonomía de Cavia aperea en el humedal del Paraná inferior

    Directory of Open Access Journals (Sweden)

    Alejandro Acosta

    2005-12-01

    Full Text Available Cavia aperea, comúnmente denominado cuis pampeano, es un pequeño roedor gregario de amplia representación en varios depósitos arqueológicos del humedal del Paraná inferior. El objetivo de este trabajo es discutir en qué medida su depósito se relaciona con factores naturales y/o antrópicos. Para ello, se evalúan distintos aspectos ecológicos y etológicos de este taxón con el fin de establecer sus posibles implicancias tafonómicas y conductuales como potencial recurso. Se estudiaron dos muestras de Cavia aperea; para el análisis se consideró su abundancia taxonómica y anatómica y diferentes aspectos tafonómicos. A su vez, su posible incorporación a la dieta es abordada desde una perspectiva teórica que la articula con la subsistencia y movilidad de las poblaciones prehispánicas del sector, cronológicamente ubicadas entre los 1.700 y 700 años AP.Cavia aperea, commonly named cuis, is a small gregarious rodent widely represented in several archaeological deposits of the lower Paraná wetlands. The aim of this paper is to discuss its presence in deposits related to natural and/or anthropic factors. Different ecological and ethological aspects of the cuis are evaluated with the purpose of establishing its taphonomic and behavioral implications as a consumed resource. Taxonomic and anatomical abundance and several taphonomic aspects were considered for the analysis of two samples of Cavia aperea. Moreover, the possible contribution of cuis to diet is linked theoretically to pre-Hispanic groups' subsistence and mobility systems (1700-700 BP.

  20. Dopaminergic differentiation of human neural stem cells mediated by co-cultured rat striatal brain slices

    DEFF Research Database (Denmark)

    Anwar, Mohammad Raffaqat; Andreasen, Christian Maaløv; Lippert, Solvej Kølvraa

    2008-01-01

    Properly committed neural stem cells constitute a promising source of cells for transplantation in Parkinson's disease, but a protocol for controlled dopaminergic differentiation is not yet available. To establish a setting for identification of secreted neural compounds promoting dopaminergic...... differentiation, we co-cultured cells from a human neural forebrain-derived stem cell line (hNS1) with rat striatal brain slices. In brief, coronal slices of neonatal rat striatum were cultured on semiporous membrane inserts placed in six-well trays overlying monolayers of hNS1 cells. After 12 days of co......-culture, large numbers of tyrosine hydroxylase (TH)-immunoreactive, catecholaminergic cells could be found underneath individual striatal slices. Cell counting revealed that up to 25.3% (average 16.1%) of the total number of cells in these areas were TH-positive, contrasting a few TH-positive cells (

  1. A novel anaerobic co-culture system for bio-hydrogen production from sugarcane bagasse.

    Science.gov (United States)

    Cheng, Jingrong; Zhu, Mingjun

    2013-09-01

    A novel co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense with pretreated sugarcane bagasse (SCB) under mild alkali conditions for bio-hydrogen production was established, exhibiting a cost-effective and synergetic advantage in bio-hydrogen production over monoculture of C. thermocellum or T. aotearoense with untreated SCB. The optimized pretreatment conditions were established to be 3% NaOH, and a liquid to solid ratio of 25:1 at 80°C for 3h. A final hydrogen production of 50.05±1.51 mmol/L was achieved with 40 g/L pretreated SCB at 55°C. The established co-culture system provides a novel consolidated bio-processing strategy for bioconversion of SCB to bio-hydrogen.

  2. Depot-dependent effects of adipose tissue explants on co-cultured hepatocytes

    DEFF Research Database (Denmark)

    Du, Zhen-Yu; Ma, Tao; Lock, Erik-Jan

    2011-01-01

    than that from epididymal ATE. However, expressions of lipolysis related genes (ATGL, HSL, perilipin-1) were higher in the epididymal adipocytes than inguinal adipocytes. Moreover, secretion of IL-6 and PGE(2) was higher from inguinal ATEs than from epididymal ATEs. There was a trend that the total...... a stronger cytotoxic response and higher level of insulin resistance in the co-cultured hepatocytes. In conclusion, our results reveal depot-dependent effects of ATEs on co-cultured primary hepatocytes, which in part may be related to a more pronounced infiltration of stromal vascular cells (SVCs......), particularly macrophages, in inguinal adipose tissue resulting in stronger responses in terms of hepatotoxicity and insulin-resistance....

  3. Infecção natural por Listeria monocytogenes em cobaios Cavia porcellus Natural infection by Listeria monocytogenes in guinea pigs (Cavia porcellus

    Directory of Open Access Journals (Sweden)

    Hugo Henrique Ferreira

    2011-04-01

    Full Text Available Listeriose foi identificada em quatro porcos-da-índia (Cavia porcellus enviados para diagnóstico postmortem. Dois cobaios (1 e 2, apresentaram especialmente nódulos esbranquiçados multifocais no fígado e ceco, alterações que corresponderam microscopicamente a múltiplos focos necróticos associados com estruturas bacterianas basofílicas, degeneração gordurosa difusa e infiltrado linfocitário periportal. Lesões similares estavam presentes no intestino delgado, ceco e baço desses dois cobaios. Os outros cobaios (3 e 4 apresentavam exclusivamente alterações pulmonares caracterizadas por coloração avermelhada difusa e áreas esbranquiçadas multifocais na superfície pleural associadas histologicamente com infiltrado neutrofílico multifocal acentuado nos lúmens alveolar e bronquiolar, além de edema interseptal e alveolar, trombos vasculares e numerosos macrófagos alveolares (pneumonia supurativa. Em seções histológicas coradas pelo Brown-Hopps, foram identificadas estruturas bacilares Gram-positivas. A avaliação imuno-histoquímica para anticorpos antiListeria monocytogenes revelou marcação fortemente positiva nos focos necróticos do fígado, ceco, baço, útero, estômago e linfonodos mesentéricos dos dois cobaios com listeriose sistêmica. Nos cobaios com pneumonia supurativa, observou-se intensa marcação nos lúmens alveolares. L. monocytogenes foi isolada de amostras de fígado e de casca de arroz utilizada como cama dos cobaios. Sugere-se que as lesões pulmonares foram consequentes à aspiração de partículas da cama de casca de arroz contaminada com L. monocytogenes.Listeriosis was identified in four guinea pigs (Cavia porcellus received for post mortem evaluation. Two animals showed multifocal white nodules in the liver and cecum. These changes corresponded microscopically to multiple necrotic foci associated with basophilic bacterial structures, diffuse lipid vacuolation, and periportal lymphocytic

  4. Production of Angkak Through Co-Culture of Monascus Purpureus and MONASCUS RUBER

    OpenAIRE

    Bibhu Prasad Panda; Saleem Javed; Mohd Ali

    2010-01-01

    Angkak (red mold rice, red yeast rice, Chinese red rice) is a traditional Chinese medicine produced by solid-state fermentation of cooked non-glutinous rice with Monascus species. The secondary metabolite of Monascus species, monacolin K /lovastatin, has been proven to lower blood lipid levels. In this study, a co-culture of Monascus purpureus MTCC 369 and Monascus ruber MTCC 1880 was used for angkak production. Four medium parameters screened by Plackett-Burman design were optimized by respo...

  5. Genome Sequence of Hypervirulent Aeromonas hydrophila Strain HZAUAH

    Science.gov (United States)

    Teng, Lin; Deng, Limei; Dong, Xingxing; Wei, Shun; Li, Jinquan

    2017-01-01

    ABSTRACT Aeromonas hydrophila, a zoonotic bacterium found in an expansive range of aquatic ecosystems, has been reported to cause severe diseases in fish, amphibians, reptiles, and mammals, including humans. Herein, we report the draft genome of the hypervirulent A. hydrophila strain HZAUAH isolated from a crucian in China. PMID:28302770

  6. Oral Challenge with Aeromonas in Protein-Malnourished Mice

    Science.gov (United States)

    1994-01-01

    precipitaion after boiling. SRBC. sheep diarrhoea (RITARD) model (25) have red blood cells (hmolysis): Yt. adrenal cell culture; CHO. Chinese hamster...did not seem to affect the susceptibility of mice to deficient substitute for colostrum was not available. Aeromonas spp. infection, but there may be

  7. Aeromonas hydrophila disturbs water and electrolyte transport in ...

    African Journals Online (AJOL)

    SERVER

    2008-02-19

    Feb 19, 2008 ... The sacs were exposed to bacteria suspension (108 cells/ ml) at 25°C for 2 h. Our results ... pathogenic, causing fish diseases. Several studies have shown that Aeromonas affects both fish and sea fruit. (Paniagua et al., 1990) ...

  8. Aeromonas spp. in het Nederlandse drinkwater; een orienterend onderzoek

    NARCIS (Netherlands)

    Versteegh JFM; During M; Havelaar AH; Koot W

    1986-01-01

    In dit rapport worden uitvoering en resultaten van een orienterend onderzoek naar het voorkomen van Aeromonas spp. in het Nederlandse drinkwater in 1985 besproken. Het bleek dat in 71% van de onderzochte monsters (rein water "af pompstation" en water uit het distributienet) in 100 ml gee

  9. SENSITIVITY OF DIFFERENT AEROMONAS SPECIES TO COPPER AND SILVER

    Science.gov (United States)

    Aeromonas bacteria are common flora in surface and ground waters and are considered to be human pathogens. They can also be found in municipally treated drinking water, likely as a component of biofilms, as found in distribution system pipes and point of use water filters. It ...

  10. Characterization of atypical Aeromonas salmonicida by different methods

    DEFF Research Database (Denmark)

    Austin, B.; Austin, D.A.; Dalsgaard, Inger;

    1998-01-01

    Fifty two isolates of atypical Aeromonas salmonicida, recovered from a wide range of hosts and geographical locations, were heterogeneous in terms of molecular and phenotypic characteristics, and represented taxa which could not be accommodated by the current classification of four subspecies...

  11. Production and properties of lipase of Aeromonas sobria.

    Science.gov (United States)

    Takahashi, Eizo; Ito, Hidenobu; Kobayashi, Hidetomo; Yamanaka, Hiroyasu; Takeda, Yoshifumi; Balakrish Nair, Gopinath; Arimoto, Sakae; Negishi, Tomoe; Okamoto, Keinosuke

    2012-05-01

    Aeromonas have been isolated from a wide variety of aquatic environments. However the number of Aeromonas in sea water is extremely small compared to that in fresh water. In in vitro culture, Aeromonas can grow in mediums containing NaCl at a concentration of 3.0%, this concentration corresponding to that of sea water. It is unclear why the number of Aeromonas is low in sea water. Exoproteins of bacteria are thought to be important for bacterial growth and survival in the environment. Previously, the present authors have shown that mediums containing 3.0% NaCl suppress production of two proteases, serine protease and metalloprotease. In this experiment, other exoproteins whose production is influenced by the amount of NaCl in the medium were analyzed. A protein whose production is repressed in medium containing 3.0% NaCl was found and purified. Biological assay of the purified protein showed that it degrades tributyrin and hydrolyzes para-nitrophenyl-fatty acylesters. These results show that the protein is a lipase. Subsequently, the nucleotide sequence of the gene encoding the lipase was determined and the amount of mRNA of the lipase gene in the cells measured. It was found that transcription of the gene is not inhibited by NaCl in the medium. This result indicates that the lipase might be synthesized, but the folding process to become an active structure does not progress smoothly in a medium containing 3.0% NaCl.

  12. Semipermeable Capsules Wrapping a Multifunctional and Self-regulated Co-culture Microenvironment for Osteogenic Differentiation.

    Science.gov (United States)

    Correia, Clara R; Pirraco, Rogério P; Cerqueira, Mariana T; Marques, Alexandra P; Reis, Rui L; Mano, João F

    2016-02-24

    A new concept of semipermeable reservoirs containing co-cultures of cells and supporting microparticles is presented, inspired by the multi-phenotypic cellular environment of bone. Based on the deconstruction of the "stem cell niche", the developed capsules are designed to drive a self-regulated osteogenesis. PLLA microparticles functionalized with collagen I, and a co-culture of adipose stem (ASCs) and endothelial (ECs) cells are immobilized in spherical liquified capsules. The capsules are coated with multilayers of poly(L-lysine), alginate, and chitosan nano-assembled through layer-by-layer. Capsules encapsulating ASCs alone or in a co-culture with ECs are cultured in endothelial medium with or without osteogenic differentiation factors. Results show that osteogenesis is enhanced by the co-encapsulation, which occurs even in the absence of differentiation factors. These findings are supported by an increased ALP activity and matrix mineralization, osteopontin detection, and the up regulation of BMP-2, RUNX2 and BSP. The liquified co-capsules also act as a VEGF and BMP-2 cytokines release system. The proposed liquified capsules might be a valuable injectable self-regulated system for bone regeneration employing highly translational cell sources.

  13. Enhanced Hydrogen Production by Co-cultures of Hydrogenase and Nitrogenase in Escherichia coli.

    Science.gov (United States)

    Lee, Hyun Jeong; Sekhon, Simranjeet Singh; Kim, Young Su; Park, Ju-Yong; Kim, Yang-Hoon; Min, Jiho

    2016-03-01

    Rhodobacter sphaeroides is a bacterium that can produce hydrogen by interaction with hydrogenase and nitrogenase. We report a hydrogen production system using co-cultivation of hydrogenase in liquid medium and immobilized nitrogenase in Escherichia coli. The recombinant plasmid has been constructed to analyze the effect of hydrogen production on the expression of hupSL hydrogenase and nifHDK nitrogenase isolated from R. sphaeroides. All recombinant E. coli strains were cultured anaerobically, and cells for nitrogenase were immobilized in agar gel, whereas cells for hydrogenase were supplemented on the nitrogenase agar gel. The hupSL hydrogenase has been observed to enhance hydrogen production and hydrogenase activity under co-culture with nifHDK nitrogenase. The maximum hydrogen production has been obtained at an agar gel concentration and a cell concentration for co-culture of 2 % and 6.4 × 10(8) CFU. Thus, co-culture of hupSL hydrogenase and nifHDK nitrogenase provides a promising route for enhancing the hydrogen production and hydrogenase activity.

  14. Flow Cytometry Sorting to Separate Viable Giant Viruses from Amoeba Co-culture Supernatants

    Science.gov (United States)

    Khalil, Jacques Y. B.; Langlois, Thierry; Andreani, Julien; Sorraing, Jean-Marc; Raoult, Didier; Camoin, Laurence; La Scola, Bernard

    2017-01-01

    Flow cytometry has contributed to virology but has faced many drawbacks concerning detection limits, due to the small size of viral particles. Nonetheless, giant viruses changed many concepts in the world of viruses, as a result of their size and hence opened up the possibility of using flow cytometry to study them. Recently, we developed a high throughput isolation of viruses using flow cytometry and protozoa co-culture. Consequently, isolating a viral mixture in the same sample became more common. Nevertheless, when one virus multiplies faster than others in the mixture, it is impossible to obtain a pure culture of the minority population. Here, we describe a robust sorting system, which can separate viable giant virus mixtures from supernatants. We tested three flow cytometry sorters by sorting artificial mixtures. Purity control was assessed by electron microscopy and molecular biology. As proof of concept, we applied the sorting system to a co-culture supernatant taken from a sample containing a viral mixture that we couldn't separate using end point dilution. In addition to isolating the quick-growing Mimivirus, we sorted and re-cultured a new, slow-growing virus, which we named “Cedratvirus.” The sorting assay presented in this paper is a powerful and versatile tool for separating viral populations from amoeba co-cultures and adding value to the new field of flow virometry. PMID:28111619

  15. Co-culture with microglia promotes neural stem cells differentiation into astrocytes

    Institute of Scientific and Technical Information of China (English)

    GU Feng; WANG Juan; FU Li; MA Yong-jie

    2011-01-01

    Background Neural stem cells (NSCs) are a self-renewing and multipotent population of the central nervous system (CNS),which are active during development and maintain homeostasis and tissue integrity throughout life.Microglias are an immune cell population resident in the CNS,which have crucial physiological functions in the developing and adult CNS.This study aimed to investigate that whether microglia co-cultured with NSCs could promote astrogliogenesis from NSCs.Methods Microglia and NSCs were co-cultured in 24-well insert plates.NSCs were plated in the bottom of the well and microglia in the insert.Fluorescent staining,Western blotting and RT-PCR were used to determine the effect of microglia on NSCs differentiation.Results Co-culture of microglia and NSCs promoted astrogliogenesis from NSCs.Several key genes,such as Notch 1,Notch 2,Notch 3,Hes 5,and NRSFwera downregulated,while the critical genes Id1 and Id2 were upregulated.BMP2 and FGF2 were upregulated.Conclusion Microglias act as a regulator of NSCs astrogliogenesis.

  16. Effects of hydrogen peroxide in a keratinocyte-fibroblast co-culture model of wound healing.

    Science.gov (United States)

    Loo, Alvin Eng Kiat; Halliwell, Barry

    2012-06-29

    Recently, there has been renewed interest in the role of reactive oxygen species (ROS), especially H(2)O(2), in wound healing. We previously showed that H(2)O(2) stimulates healing in a keratinocyte scratch wound model. In this paper, we used a more complex and physiologically relevant model that involves co-culturing primary keratinocytes and fibroblasts. We found that the two main cell types within the skin have different sensitivities to H(2)O(2) and to the widely used "antioxidant"N-acetyl-l-cysteine (NAC). Keratinocytes were very resistant to the toxicity of H(2)O(2) (250 and 500 μM) or NAC (5 mM). However, the viability of fibroblasts was decreased by both compounds. Using the co-culture model, we also found that H(2)O(2) increases re-epithelialization while NAC retards it. Our data further illustrate the possible role of ROS in wound healing and the co-culture model should be useful for screening agents that may influence the wound healing process. Copyright © 2012 Elsevier Inc. All rights reserved.

  17. Semipermeable Capsules Wrapping a Multifunctional and Self-regulated Co-culture Microenvironment for Osteogenic Differentiation

    Science.gov (United States)

    Correia, Clara R.; Pirraco, Rogério P.; Cerqueira, Mariana T.; Marques, Alexandra P.; Reis, Rui L.; Mano, João F.

    2016-02-01

    A new concept of semipermeable reservoirs containing co-cultures of cells and supporting microparticles is presented, inspired by the multi-phenotypic cellular environment of bone. Based on the deconstruction of the “stem cell niche”, the developed capsules are designed to drive a self-regulated osteogenesis. PLLA microparticles functionalized with collagen I, and a co-culture of adipose stem (ASCs) and endothelial (ECs) cells are immobilized in spherical liquified capsules. The capsules are coated with multilayers of poly(L-lysine), alginate, and chitosan nano-assembled through layer-by-layer. Capsules encapsulating ASCs alone or in a co-culture with ECs are cultured in endothelial medium with or without osteogenic differentiation factors. Results show that osteogenesis is enhanced by the co-encapsulation, which occurs even in the absence of differentiation factors. These findings are supported by an increased ALP activity and matrix mineralization, osteopontin detection, and the up regulation of BMP-2, RUNX2 and BSP. The liquified co-capsules also act as a VEGF and BMP-2 cytokines release system. The proposed liquified capsules might be a valuable injectable self-regulated system for bone regeneration employing highly translational cell sources.

  18. (13)C-metabolic flux analysis of co-cultures: A novel approach.

    Science.gov (United States)

    Gebreselassie, Nikodimos A; Antoniewicz, Maciek R

    2015-09-01

    In this work, we present a novel approach for performing (13)C metabolic flux analysis ((13)C-MFA) of co-culture systems. We demonstrate for the first time that it is possible to determine metabolic flux distributions in multiple species simultaneously without the need for physical separation of cells or proteins, or overexpression of species-specific products. Instead, metabolic fluxes for each species in a co-culture are estimated directly from isotopic labeling of total biomass obtained using conventional mass spectrometry approaches such as GC-MS. In addition to determining metabolic fluxes, this approach estimates the relative population size of each species in a mixed culture and inter-species metabolite exchange. As such, it enables detailed studies of microbial communities including species dynamics and interactions between community members. The methodology is experimentally validated here using a co-culture of two E. coli knockout strains. Taken together, this work greatly extends the scope of (13)C-MFA to a large number of multi-cellular systems that are of significant importance in biotechnology and medicine.

  19. Streamlining gene expression analysis: integration of co-culture and mRNA purification.

    Science.gov (United States)

    Berry, Scott M; Singh, Chandresh; Lang, Jessica D; Strotman, Lindsay N; Alarid, Elaine T; Beebe, David J

    2014-02-01

    Co-culture of multiple cell types within a single device enables the study of paracrine signaling events. However, extracting gene expression endpoints from co-culture experiments is laborious, due in part to pre-PCR processing of the sample (i.e., post-culture cell sorting and nucleic acid purification). Also, a significant loss of nucleic acid may occur during these steps, especially with microfluidic cell culture where lysate volumes are small and difficult to access. Here, we describe an integrated platform for performing microfluidic cell culture and extraction of mRNA for gene expression analysis. This platform was able to recover 30-fold more mRNA than a similar, non-integrated system. Additionally, using a breast cancer/bone marrow stroma co-culture, we recapitulated stromal-dependent, estrogen-independent growth of the breast cancer cells, coincident with transcriptional changes. We anticipate that this platform will be used for streamlined analysis of paracrine signaling events as well as for screening potential drugs and/or patient samples.

  20. Co-culturing Effects of Coexisting Bacteria on Wood Degradation by Trametes versicolor.

    Science.gov (United States)

    Kamei, Ichiro

    2017-01-01

    White-rot fungi are the main decomposers of wood cell-wall polymer in forest ecosystems. Little is known, however, about the interactions between white-rot fungi and other coexisting microorganisms in decayed wood. A white-rot fungus, Trametes versicolor strain TN6F, was isolated from a fruit body, and 44 strains of coexisting cultivable bacteria were isolated from its substrate, natural white rot-decayed wood. The effects of these bacteria on fungal growth were examined by an in vitro confrontation growth assay. Among the isolates, nine bacterial strains inhibited the growth of strain TN6F, while 35 strains did not affect the growth of TN6F. However, when co-cultured with strain TN6F on wood powder, many bacterial strains promoted the weight loss of the substrate. A subsequent chemical composition analysis showed that co-culturing accelerated delignification. Higher laccase activity was detected when strain TN6F was co-cultured on wood powder medium with bacterial strains TN6W-26 or TN6W-27. These results indicate that some bacterial strains might promote wood degradation.

  1. The improved PCR of the fstA (ferric siderophore receptor) gene differentiates the fish pathogen Aeromonas salmonicida from other Aeromonas species.

    Science.gov (United States)

    Beaz-Hidalgo, Roxana; Latif-Eugenín, Fadua; Figueras, María José

    2013-10-25

    The members of the genus Aeromonas are autochthonous of aquatic ecosystems and several species have been associated to septicaemia, ulcerative and haemorrhagic diseases in fish, causing significant mortality in both wild and farmed, freshwater and marine fish species. The species Aeromonas salmonicida is generally recognized as the most important fish pathogen responsible for epidemic outbreaks of furunculosis in salmonids, also being able to produce infections in other cultured fish such as turbot, halibut, sea bream or goldfish. New species, i.e. Aeromonas aquariorum, Aeromonas tecta and Aeromonas piscicola, have recently been discovered and isolated from diseased fish. The species A. piscicola and Aeromonas bestiarum are practically impossible to differentiate phenotypically and genetically (when using the 16S rRNA gene) from each other and from A. salmonicida. In the present study, two previously described PCR protocols, based on the fstA and gyrB genes, for the specific detection of A. salmonicida were re-evaluated with the type strains of all Aeromonas species and with a set of A. piscicola and A. bestiarum strains. Contrary to what had been published previously it was demonstrated that the gyrB-PCR is not specific for A. salmonicida because of cross-reactions with other Aeromonas species. However, in agreement with previous results, A. salmonicida was detected on the basis of the fstA-PCR, for which an improved protocol was proposed.

  2. Spirulina chitosan gel induction on healing process of Cavia cobaya post extraction socket

    Directory of Open Access Journals (Sweden)

    Rostiny Rostiny

    2014-03-01

    Full Text Available Background: Prominent residual ridge is necessary to gain retention and stabilility for succesful prosthodontic treatment such as removable, fixed or implant. Spirulina is a natural substance that can help tissue healing and chitosan also a natural substance that reported to have the ability to help bone remodelling. The combination gel of spirulina and chitosan could be considered as an alternative material to maintain residual ridge height after tooth extraction. Purpose: The aim of study was to examine the effect of combination gel of Spirulina and chitosan on healing process of Cavia cobaya post tooth extraction socket by counting the amount of osteoclast, osteoblast and colagen as an indicator. Methods: Twenty eight cavia cobaya were divided into 4 groups. Insisive mandible extraction was done and the sockets were filled with 3% CMCNa for control groups, 3% spirulina chitosan 200 mg for group 1, 6% spirulina chitosan 200 mg for group 2, 12% spirulina chitosan 200 mg for group 3. After 30 days, histopathology examination was done by using microscope to count the amount of osteoclast, osteoblast and collagen. Results: Data was analyzed by using Anova and Tukey HSD. For osteoclast, there was no significant different between every groups, while for osteoblast and collagen there was significant different between groups. The results showed that induction of combination gel spirulina chitosan was able to accumulate collagen fiber and resulting faster wound healing. Conclusion: Combination 12% gel spirulina chitosan 200 mg could be used as an alternative material for better bone remodeling after tooth extraction.Latar belakang: Residual ridge yang prominen sangat dibutuhkan untuk mendapatkan retensi dan stabilitas untuk menunjang keberhasilan perawatan di bidang prostodonsia seperti pada kasus removable, fixed atau implant. Tindakan pencabutan gigi dapat merusak jaringan periodontal, sementum dan tulang alveolar yang mengakibatkan resorbsi ridge

  3. Interactions between airway epithelial cells and dendritic cells during viral infections using an in vitro co-culture model

    Science.gov (United States)

    Rationale: Historically, single cell culture models have been limited in pathological and physiological relevance. A co-culture model of dendritic cells (DCs) and differentiated human airway epithelial cells was developed to examine potential interactions between these two cell t...

  4. Biodegradation of crude oil by a defined co-culture of indigenous bacterial consortium and exogenous Bacillus subtilis.

    Science.gov (United States)

    Tao, Kaiyun; Liu, Xiaoyan; Chen, Xueping; Hu, Xiaoxin; Cao, Liya; Yuan, Xiaoyu

    2017-01-01

    The aim of this work was to study biodegradation of crude oil by defined co-cultures of indigenous bacterial consortium and exogenous Bacillus subtilis. Through residual oil analysis, it is apparent that the defined co-culture displayed a degradation ratio (85.01%) superior to indigenous bacterial consortium (71.32%) after 7days of incubation when ratio of inoculation size of indigenous bacterial consortium and Bacillus subtilis was 2:1. Long-chain n-alkanes could be degraded markedly by Bacillus subtilis. Result analysis of the bacterial community showed that a decrease in bacterial diversity in the defined co-culture and the enrichment of Burkholderiales order (98.1%) degrading hydrocarbons. The research results revealed that the promising potential of the defined co-culture for application to degradation of crude oil.

  5. Antibiotic resistance in Aeromonas upstream and downstream of a water resource recovery facility.

    Science.gov (United States)

    Cisar, Cindy R; Henderson, Samantha K; Askew, Maegan L; Risenhoover, Hollie G; McAndrews, Chrystle R; Kennedy, S Dawn; Paine, C Sue

    2014-09-01

    Aeromonas strains isolated from sediments upstream and downstream of a water resource recovery facility (WRRF) over a two-year time period were tested for susceptibility to 13 antibiotics. Incidence of resistance to antibiotics, antibiotic resistance phenotypes, and diversity (based on resistance phenotypes) were compared in the two populations. At the beginning of the study, the upstream and downstream Aeromonas populations were different for incidence of antibiotic resistance (p resistance phenotypes (p antibiotic resistance in Aeromonas in stream sediments fluctuates considerably over time and (2) suggest that WRRF effluent does not, when examined over the long- term, affect antibiotic resistance in Aeromonas in downstream sediment.

  6. Incidence of Aeromonas bacteremia in Southern Taiwan: vibrio and Salmonella bacteremia as comparators.

    Science.gov (United States)

    Wu, Chi-Jung; Chen, Po-Lin; Tang, Hung-Jen; Chen, Hung-Mo; Tseng, Fan-Chen; Shih, Hsin-I; Hung, Yuan-Pin; Chung, Chih-Huan; Ko, Wen-Chien

    2014-04-01

    The aim of the investigation was to describe the incidence of Aeromonas bacteremia in a city with a population of about 1.87 million inhabitants, located in southern Taiwan, between 2008 and 2010. Such data were compared with the incidences of Vibrio and Salmonella bacteremia in the same period and the incidence of Aeromonas bacteremia in other countries in the literature. The data revealed the average annual incidences of bacteremia due to Aeromonas, Vibrio, and Salmonella species were 76, 38, and 103 cases/million inhabitants, respectively. The incidence of Aeromonas bacteremia was higher than those in Western countries.

  7. Interaction of Aeromonas strains with lactic acid bacteria via Caco-2 cells.

    Science.gov (United States)

    Hatje, E; Neuman, C; Katouli, M

    2014-01-01

    The genus Aeromonas includes some species that have now been identified as human pathogens of significant medical importance. We investigated the ability of 13 selected Aeromonas strains belonging to nine species isolated from clinical cases (n = 5), environmental waters (n = 5), and fish (n = 3) to adhere to and translocate Caco-2 cells in the absence and presence of two lactic acid bacteria (LAB), i.e., Lactobacillus acidophilus and Bifidobacterium breve. Aeromonas isolates were also assessed for their cytotoxicity, the presence of virulence genes, and hemolysin production. Among the clinical isolates, one strain of Aeromonas veronii biovar veronii and two strains of Aeromonas hydrophila carried cytotoxin (act), heat-labile toxin (alt), hemolysin (hlyA), and aerolysin (aerA) genes, were cytotoxic to Vero cells, produced hemolysin, and showed higher adherence to Caco-2 cells. In contrast, this was seen in only one environmental strain, a strain of A. veronii biovar sobria. When Aeromonas strains were coinoculated with LAB onto Caco-2 cells, their level of adhesion was reduced. However, their rate of translocation in the presence of LAB increased and was significantly (P Aeromonas and LAB strains could have a detrimental effect on the Caco-2 cells, allowing the Aeromonas to translocate more readily, or the presence of the LAB stimulated the Aeromonas strains to produce more toxins and/or increase their translocation rate.

  8. Hyaline cartilage tissue is formed through the co-culture of passaged human chondrocytes and primary bovine chondrocytes.

    Science.gov (United States)

    Taylor, Drew W; Ahmed, Nazish; Hayes, Anthony J; Ferguson, Peter; Gross, Allan E; Caterson, Bruce; Kandel, Rita A

    2012-08-01

    To circumvent the problem of a sufficient number of cells for cartilage engineering, the authors previously developed a two-stage culture system to redifferentiate monolayer culture-expanded dedifferentiated human articular chondrocytes by co-culture with primary bovine chondrocytes (bP0). The aim of this study was to analyze the composition of the cartilage tissue formed in stage 1 and compare it with bP0 grown alone to determine the optimal length of the co-culture stage of the system. Biochemical data show that extracellular matrix accumulation was evident after 2 weeks of co-culture, which was 1 week behind the bP0 control culture. By 3 to 4 weeks, the amounts of accumulated proteoglycans and collagens were comparable. Expression of chondrogenic genes, Sox 9, aggrecan, and collagen type II, was also at similar levels by week 3 of culture. Immunohistochemical staining of both co-culture and control tissues showed accumulation of type II collagen, aggrecan, biglycan, decorin, and chondroitin sulfate in appropriate zonal distributions. These data indicate that co-cultured cells form cartilaginous tissue that starts to resemble that formed by bP0 after 3 weeks, suggesting that the optimal time to terminate the co-culture stage, isolate the now redifferentiated cells, and start stage 2 is just after 3 weeks.

  9. Comparative study of nickel resistance of pure culture and co-culture of Acidithiobacillus thiooxidans and Leptospirillum ferriphilum.

    Science.gov (United States)

    Xu, Ying; Yin, Huaqun; Jiang, Huidan; Liang, Yili; Guo, Xue; Ma, Liyuan; Xiao, Yunhua; Liu, Xueduan

    2013-09-01

    The effect of Ni²⁺ on the growth and functional gene expression of the pure culture and co-culture of Acidithiobacillus thiooxidans and Leptospirillum ferriphilum has been studied. Compared with the pure culture, the co-culture showed a stronger sulfur and ferrous ion oxidation activity. At 100 mM, A. thiooxidans in co-culture grew faster and had 48 h shorter lag phases. The cell number of A. thiooxidans in co-culture was about 5 times higher than that in pure culture. The existence of A. thiooxidans in co-culture activated the expression of some metal resistance genes in L. ferriphilum at least 16 h in advance. A. thiooxidans in co-culture tends to chose more efficient pathways to transport nickel ion, ensuring the export of heavy metal was faster and more effective than that in pure culture. All the data indicated that there were synergetic interactions between iron- and sulfur-oxidizing bacteria under the stress of Ni²⁺.

  10. Co-culture with periodontal ligament stem cells enhances osteogenic gene expression in de-differentiated fat cells.

    Science.gov (United States)

    Tansriratanawong, Kallapat; Tamaki, Yuichi; Ishikawa, Hiroshi; Sato, Soh

    2014-10-01

    In recent decades, de-differentiated fat cells (DFAT cells) have emerged in regenerative medicine because of their trans-differentiation capability and the fact that their characteristics are similar to bone marrow mesenchymal stem cells. Even so, there is no evidence to support the osteogenic induction using DFAT cells in periodontal regeneration and also the co-culture system. Consequently, this study sought to evaluate the DFAT cells co-culture with periodontal ligament stem cells (PDLSCs) in vitro in terms of gene expression by comparing runt-related transcription factor 2 (RUNX2) and Peroxisome proliferator-activated receptor gamma 2 (PPARγ2) genes. We isolated DFAT cells from mature adipocytes and compared proliferation with PDLSCs. After co-culture with PDLSCs, we analyzed transcriptional activity implying by DNA methylation in all adipogenic gene promoters using combined bisulfite restriction analysis. We compared gene expression in RUNX2 gene with the PPARγ2 gene using quantitative RT-PCR. After being sub-cultured, DFAT cells demonstrated morphology similar to fibroblast-like cells. At the same time, PDLSCs established all stem cell characteristics. Interestingly, the co-culture system attenuated proliferation while enhancing osteogenic gene expression in RUNX2 gene. Using the co-culture system, DFAT cells could trans-differentiate into osteogenic lineage enhancing, but conversely, their adipogenic characteristic diminished. Therefore, DFAT cells and the co-culture system might be a novel cell-based therapy for promoting osteogenic differentiation in periodontal regeneration.

  11. Effects of hydroxybenzyl alcohols on melanogenesis in melanocyte-keratinocyte co-culture and monolayer culture of melanocytes.

    Science.gov (United States)

    Liu, Szu-Hsiu; Chu, I-Ming; Pan, I-Horng

    2008-08-01

    In mammalian skin, melanocyte proliferation and melanogenesis can be stimulated by keratinocytes, fibroblasts and other regulatory factors. To determine whether hydroxybenzyl alcohols (HBAs) show more inhibitory in melanocytes cultured alone or in melanocytes co-cultured with keratinocytes, we developed a murine melanocyte-keratinocyte co-culture model to investigate the pigmentation regulators in company with other melanogenic inhibitors and stimulators. It was found that the effects of HBAs and melanogenic factors were more evident in melanocytes co-cultured with keratinocytes. Keratinocytes may play a synergistic role in melanocyte melanogenesis and influence the pigment production. The tests in the co-culture model also imply that the inhibitory effects of HBAs on melanogenesis are due to the direct inhibition of melanosomal tyrosinase activity. HBAs showed a low cytotoxicity. The eventual results proved that HBAs are promising and safe agents for skin whitening in melanocyte alone and in co-culture systems. The co-culture model provides a more physiologically realistic condition to study the interaction between melanocytes and keratinocytes, which enables a reliable screening system for depigmenting compounds.

  12. A three-dimensional co-culture model of the aortic valve using magnetic levitation.

    Science.gov (United States)

    Tseng, Hubert; Balaoing, Liezl R; Grigoryan, Bagrat; Raphael, Robert M; Killian, T C; Souza, Glauco R; Grande-Allen, K Jane

    2014-01-01

    The aortic valve consists of valvular interstitial cells (VICs) and endothelial cells (VECs). While these cells are understood to work synergistically to maintain leaflet structure and valvular function, few co-culture models of these cell types exist. In this study, aortic valve co-cultures (AVCCs) were assembled using magnetic levitation and cultured for 3 days. Immunohistochemistry and quantitative reverse-transcriptase polymerase chain reaction were used to assess the maintenance of cellular phenotype and function, and the formation of extracellular matrix. AVCCs stained positive for CD31 and α-smooth muscle actin (αSMA), demonstrating that the phenotype was maintained. Functional markers endothelial nitric oxide synthase (eNOS), von Willebrand factor (VWF) and prolyl-4-hydroxylase were present. Extracellular matrix components collagen type I, laminin and fibronectin also stained positive, with reduced gene expression of these proteins in three dimensions compared to two dimensions. Genes for collagen type I, lysyl oxidase and αSMA were expressed less in AVCCs than in 2-D cultures, indicating that VICs are quiescent. Co-localization of CD31 and αSMA in the AVCCs suggests that endothelial-mesenchymal transdifferentiation might be occurring. Differences in VWF and eNOS in VECs cultured in two and three dimensions also suggests that the AVCCs possibly have anti-thrombotic potential. Overall, a co-culture model of the aortic valve was designed, and serves as a basis for future experiments to understand heart valve biology. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  13. Enhanced Keratinocyte Proliferation and Migration in Co-culture with Fibroblasts

    Science.gov (United States)

    Wang, Zhenxiang; Wang, Ying; Farhangfar, Farhang; Zimmer, Monica; Zhang, Yongxin

    2012-01-01

    Wound healing is primarily controlled by the proliferation and migration of keratinocytes and fibroblasts as well as the complex interactions between these two cell types. To investigate the interactions between keratinocytes and fibroblasts and the effects of direct cell-to-cell contact on the proliferation and migration of keratinocytes, keratinocytes and fibroblasts were stained with different fluorescence dyes and co-cultured with or without transwells. During the early stage (first 5 days) of the culture, the keratinocytes in contact with fibroblasts proliferated significantly faster than those not in contact with fibroblasts, but in the late stage (11th to 15th day), keratinocyte growth slowed down in all cultures unless EGF was added. In addition, keratinocyte migration was enhanced in co-cultures with fibroblasts in direct contact, but not in the transwells. Furthermore, the effects of the fibroblasts on keratinocyte migration and growth at early culture stage correlated with heparin-binding EGF-like growth factor (HB-EGF), IL-1α and TGF-β1 levels in the cultures where the cells were grown in direct contact. These effects were inhibited by anti-HB-EGF, anti-IL-1α and anti-TGF-β1 antibodies and anti-HB-EGF showed the greatest inhibition. Co-culture of keratinocytes and IL-1α and TGF-β1 siRNA-transfected fibroblasts exhibited a significant reduction in HB-EGF production and keratinocyte proliferation. These results suggest that contact with fibroblasts stimulates the migration and proliferation of keratinocytes during wound healing, and that HB-EGF plays a central role in this process and can be up-regulated by IL-1α and TGF-β1, which also regulate keratinocyte proliferation differently during the early and late stage. PMID:22911722

  14. Enhanced keratinocyte proliferation and migration in co-culture with fibroblasts.

    Directory of Open Access Journals (Sweden)

    Zhenxiang Wang

    Full Text Available Wound healing is primarily controlled by the proliferation and migration of keratinocytes and fibroblasts as well as the complex interactions between these two cell types. To investigate the interactions between keratinocytes and fibroblasts and the effects of direct cell-to-cell contact on the proliferation and migration of keratinocytes, keratinocytes and fibroblasts were stained with different fluorescence dyes and co-cultured with or without transwells. During the early stage (first 5 days of the culture, the keratinocytes in contact with fibroblasts proliferated significantly faster than those not in contact with fibroblasts, but in the late stage (11(th to 15(th day, keratinocyte growth slowed down in all cultures unless EGF was added. In addition, keratinocyte migration was enhanced in co-cultures with fibroblasts in direct contact, but not in the transwells. Furthermore, the effects of the fibroblasts on keratinocyte migration and growth at early culture stage correlated with heparin-binding EGF-like growth factor (HB-EGF, IL-1α and TGF-β1 levels in the cultures where the cells were grown in direct contact. These effects were inhibited by anti-HB-EGF, anti-IL-1α and anti-TGF-β1 antibodies and anti-HB-EGF showed the greatest inhibition. Co-culture of keratinocytes and IL-1α and TGF-β1 siRNA-transfected fibroblasts exhibited a significant reduction in HB-EGF production and keratinocyte proliferation. These results suggest that contact with fibroblasts stimulates the migration and proliferation of keratinocytes during wound healing, and that HB-EGF plays a central role in this process and can be up-regulated by IL-1α and TGF-β1, which also regulate keratinocyte proliferation differently during the early and late stage.

  15. Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture

    Directory of Open Access Journals (Sweden)

    Restu Syamsul Hadi

    2015-12-01

    Full Text Available BACKGROUND Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC. METHODS In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4 expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1 proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. RESULTS Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05. Calculated viable cell numbers increased 1.8 and 5.1- fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity CONCLUSIONS Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.

  16. Allogeneic human dermal fibroblasts are viable in peripheral blood mononuclear co-culture

    Directory of Open Access Journals (Sweden)

    Restu Syamsul Hadi

    2014-08-01

    Full Text Available Background Transplanted allogeneic dermal fibroblasts retain stem cell subpopulations, and are easily isolated, expanded and stored using standard techniques. Their potential for regenerative therapy of chronic wounds should be evaluated. The aim of this study was to determine allogeneic fibroblast viability in the presence of peripheral blood mononuclear cells (PBMC. Methods In this experimental study, fibroblasts were isolated from foreskin explants, expanded in the presence of serum, and stored using slow-freezing. We used one intervention group of allogeneic fibroblasts co-cultured with PBMC and 2 control groups of separate fibroblast and PBMC cultures.Fibroblasts were characterized by their collagen secretion and octamer-binding transcription factor 4 (OCT4 expression. Viability was evaluated using water soluble tetrazolium-1 (WST-1 proliferation assay. Absorbances were measured at 450 nm. Data analysis was performed by student’s paired t-test. Results Dermal fibroblasts were shown to secrete collagen, express OCT4, be recoverable after cryopreservation, and become attached to the culture dish in a co-culture with PBMC. Co-cultured and control fibroblasts had no significantly different cell viabilities (p>0.05. Calculated viable cell numbers increased 1.8 and 5.1-fold, respectively, at days 2 and 4 in vitro. Both groups showed comparable doubling times at days 2 and 4 in vitro. PBMC did not interfere with allogeneic fibroblast viability and proliferative capacity Conclusions Allogeneic fibroblasts remain viable and proliferate in the presence of host PBMC. Future research should evaluate allogeneic human dermal fibroblast competency in clinical settings. Dermal fibroblasts are a potential source for cell therapy in chronic wound management.

  17. Generation of co-culture spheroids as vascularisation units for bone tissue engineering

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    R Walser

    2013-11-01

    Full Text Available Cell spheroids represent attractive building units for bone tissue engineering, because they provide a three-dimensional environment with intensive direct cell-cell contacts. Moreover, they allow for co-culture of both osteoblasts and vessel-forming cells, which may markedly increase their survival and vascularisation after transplantation. To test this hypothesis, we generated co-culture spheroids by aggregating different combinations of primary human osteoblasts (HOB, human dermal microvascular endothelial cells (HDMEC and normal human dermal fibroblasts (NHDF using the liquid overlay technique. Mono-culture spheroids consisting either of HOB or HDMEC served as controls. After in vitro characterisation, the different spheroids were transplanted into dorsal skinfold chambers of CD1 nu/nu mice to study in vivo their viability and vascularisation over a 2-week observation period by means of repetitive intravital fluorescence microscopy and immunohistochemistry. In vitro, co-culture spheroids containing HDMEC rapidly formed dense tubular vessel-like networks within 72 h and exhibited a significantly decreased rate of apoptotic cell death when compared to mono-culture HDMEC spheroids. After transplantation, these networks interconnected to the host microvasculature by external inosculation. Of interest, this process was most pronounced in HOB-HDMEC spheroids and could not further be improved by the addition of NHDF. Accordingly, HOB-HDMEC spheroids were larger when compared to the other spheroid types. These findings indicate that HOB-HDMEC spheroids exhibit excellent properties to preserve viability and to promote proliferation and vascularisation. Therefore, they may be used as functional vascularisation units in bone tissue engineering for the seeding of scaffolds or for the vitalisation of non-healing large bone defects.

  18. Atrazine degradation by fungal co-culture enzyme extracts under different soil conditions.

    Science.gov (United States)

    Chan-Cupul, Wilberth; Heredia-Abarca, Gabriela; Rodríguez-Vázquez, Refugio

    2016-01-01

    This investigation was undertaken to determine the atrazine degradation by fungal enzyme extracts (FEEs) in a clay-loam soil microcosm contaminated at field application rate (5 μg g(-1)) and to study the influence of different soil microcosm conditions, including the effect of soil sterilization, water holding capacity, soil pH and type of FEEs used in atrazine degradation through a 2(4) factorial experimental design. The Trametes maxima-Paecilomyces carneus co-culture extract contained more laccase activity and hydrogen peroxide (H2O2) content (laccase = 18956.0 U mg protein(-1), H2O2 = 6.2 mg L(-1)) than the T. maxima monoculture extract (laccase = 12866.7 U mg protein(-1), H2O2 = 4.0 mg L(-1)). Both extracts were able to degrade atrazine at 100%; however, the T. maxima monoculture extract (0.32 h) achieved a lower half-degradation time than its co-culture with P. carneus (1.2 h). The FEE type (p = 0.03) and soil pH (p = 0.01) significantly affected atrazine degradation. The best degradation rate was achieved by the T. maxima monoculture extract in an acid soil (pH = 4.86). This study demonstrated that both the monoculture extracts of the native strain T. maxima and its co-culture with P. carneus can efficiently and quickly degrade atrazine in clay-loam soils.

  19. Allelopathy in a leguminous mangrove plant, Derris indica: protoplast co-culture bioassay and rotenone effect.

    Science.gov (United States)

    Inoue, Aya; Mori, Daisuke; Minagawa, Reiko; Fujii, Yoshiharu; Sasamoto, Hamako

    2015-05-01

    To investigate allelopathic activity of a leguminous mangrove plant, Derris indica, the 'Protoplasts Co-culture Method' for bioassay of allelopathy was developed using suspension culture. A suspension culture was induced from immature seed and sub-cultured in Murashige and Skoog's (MS) basal medium containing 10 μM each of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA). The protoplasts were isolated using the separate wells method with 2% each of Cellulase RS, Driselase 20 and Macerozyme R10 in 0.4 M mannitol solution. Protoplast cultures of D. indica revealed that high concentrations of cytokinins, BA and thidiazuron, were effective for cell divisions. The co-cultures of D. indica protoplasts with recipient lettuce protoplasts using 96 multi-well culture plates were performed in MS basal medium containing 0.4 M mannitol solution and 1 μM 2,4-D and 0.1 μM BA. The protoplast density of D. indica used in co-culturing varied from 6 x 10(3) - 10(5) / mL. Very strong inhibitory allelopathic effects of D. indica protoplasts on lettuce protoplast growth were found. A similar strong inhibitory allelopathic activity of dried young leaves on lettuce seedling growth was also observed by using the sandwich method. Rotenone, which is a component of Derris root, dissolved in DMSO, was highly inhibitory on the growth of lettuce protoplasts in culture and this could be one of the causes of the strong allelopathic activity of D. indica.

  20. L’immunogenicità nella cavia e nel cavallo di due formulazioni di un vaccino inattivato e adiuvato per la peste equina

    Directory of Open Access Journals (Sweden)

    Gaetano Federico Ronchi

    2012-03-01

    Full Text Available L’efficacia di due vaccini monovalenti, inattivati e adiuvati per il controllo della Peste Equina, allestiti con i sierotipi 5 e 9, è stata saggiata su cavia per selezionare la formulazione con le migliori capacità immunogene. Nella formulazione dei vaccini sono state prese in considerazione: la risposta immunitaria evocata nella cavia e le proprietà infiammatorie di due diversi tipi di adiuvanti precedentemente saggiati nella specie di destino del vaccino.Il vaccino allestito con il sierotipo 9, saggiato in uno studio pilota su cavallo, si è dimostrato capace fin dalla prima somministrazione di stimolare la produzione di anticorpi neutralizzanti. La risposta anticorpale evocata ha subito un marcato rialzo dopo la somministrazione della dose di richiamo, effettuata dopo 28 giorni, perdurando per almeno 10 mesi. La cavia sembra essere un utile modello di laboratorio per la valutazione delle proprietà antigeniche dei vaccini contro la peste equina.

  1. A 3D co-culture microtissue model of the human placenta for nanotoxicity assessment

    DEFF Research Database (Denmark)

    Muoth, Carina; Wichser, Adrian; Monopoli, Marco;

    2016-01-01

    and functionality of the placental tissue. The effects of NPs on the human placenta are not well studied or understood, and predictive in vitro placenta models to achieve mechanistic insights on NP-placenta interactions are essentially lacking. Using the scaffold-free hanging drop technology, we developed a well-organized...... and highly reproducible 3D co-culture microtissue (MT) model consisting of a core of placental fibroblasts surrounded by a trophoblast cell layer, which resembles the structure of the in vivo placental tissue. We could show that secretion levels of human chorionic gonadotropin (hCG) were significantly higher...

  2. Measuring Contagion of Subprime Crisis Based on MVMQ-CAViaR Method

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    Wuyi Ye

    2014-01-01

    Full Text Available The analysis of financial contagion is a topical issue in international finance and portfolio management. In this paper, we investigate whether the global financial crisis originating from American subprime crisis spreads to China, Japan, UK, France, and Germany. Firstly, multivariate conditional autoregressive value at risk (MV-CAViaR models are applied to the whole sample to analyze the variation of market risk among these countries. By dividing the sampling period into three important subperiods (precrisis period, crisis period, and recovery period, we examine the changes of the dependence structure of risk during each period. Comparing with the situations in precrisis period, if the estimated coefficients become significant or market risk increases during the crisis, it implies the existence of contagion from the angle of coefficient significance or risk. The findings show that the concerned coefficients are significant or the market risks of the tested countries increase during the crisis except for China. The results imply that there is contagion from the US to all other countries, except for China. Furthermore, the changes of the market risk are found to be consistent with market events and media reports during that period.

  3. Study on postpartum estrus of guinea pigs (Cavia cobaya using Anredera cordifolia leaf extract

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    D. Wijayanti

    2017-04-01

    Full Text Available Aim: The purpose of this study was to determine the postpartum estrus cycle of guinea pigs (Cavia cobaya using Anredera cordifolia leaf extract. Materials and Methods: Materials used were 8 males and 8 females of C. cobaya with body weight ranged 400-450 g. Mating ratio applied was 1:1. Treatments given were 0, 10, 50 and 90 mg of A. cordifolia leaf extract/head, designated as T0, T1, T2 and T3, respectively. Two females were subjected for each treatment. A. cordifolia leaf extract was administered orally from 10 days prepartum to 10 days postpartum. Observation of mating behavior, vulva morphology, and vaginal smear preparation was done in the afternoon for 10 days subsequence postpartum. Data were analyzed by univariate method and descriptively. Results: The results showed that the addition of A. cordifolia leaf extract 50 mg orally could accelerate the time of postpartum estrus based on the average frequency of mating behavior consisting of behavioral approach, allow the buck to sniffing her, mating positions, standing heat, lordosis position, and copulation. During estrus, vulva morphology was red color, had much mucus and no thin membrane covering vagina. There were a lot of superficial cells on vagina. Conclusion: The best treatment to accelerate occurring postpartum estrus was the addition of A. cordifolia leaf extract as many as 50 mg/head weight (T2 orally.

  4. Anatomy of the heart of Guinea pig fetuses in late gestation (Cavia porcellus [Linnaeus, 1758

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    Mariangela de Toledo Barbino

    2011-09-01

    Full Text Available This study has been developed in order to describe the anatomy of the heart of Guinea pig (Cavia porcellus [Linnaeu,1758] fetuses in late gestation and to obtain anatomical characteristics that can support and help the understanding of the physiology of fetal circulation. Fetuses have been collected from three female in late gestation, which were described and photographed macroscopically, trhough light microscopy in HE and scanning electronic microscopy. The fetal heart is divided into four cavities, it is surrounded by the pericardium, and it has three layers: epicardium, myocardium, and endocardium. Foramen ovale has a great importance in the fetal circulation, but it could not be visualized in sections carried out in this study. Through these results we conclude that the hearts of Guinea pig fetuses differ from those of other species in structures such as the cranial vena cava, which is not divided into left and right, and that the walls thickness of left and right ventricles does not differ significantly.

  5. Anatomy of the liver of Guinea pig fetuses in late gestation (Cavia porcellus [Linnaeus, 1758

    Directory of Open Access Journals (Sweden)

    Mariangela de Toledo Barbino

    2011-09-01

    Full Text Available To describe the anatomy of the liver of Guinea pig (Cavia porcellus [Linnaeus, 1758] fetuses in late gestation and to obtain anatomical characteristics that can support and help the understanding of the physiology of fetal circulation. Three fetuses of Guinea pig in late gestation have been used, which were dissected and had their livers removed. These were analized, described, and photographed macroscopically and microscopically through light microscopy in HE and scanning electronic microscopy. Macroscopically, the fetuses livers have a reddish brown color and their division into lobes is clearly seen, as well as the presence of the gallbladder. The liver is divided into left lateral lobe, left medial lobe, right lateral lobe, right medial lobe, quadrate lobe, and caudate lobe with caudate and papillary processes. Through light microscopy, highly vascularized tissue is observed, and the left portion of the liver receive blood from the placenta through the umbilical vein, and the right portion is irrigated by the portal vein. The structures found on the liver of Guinea pig fetuses in late gestation are anatomically similar to those of other mammalian species.

  6. Investigation Into the Humaneness of Slaughter Methods for Guinea Pigs (Cavia porcelus) in the Andean Region.

    Science.gov (United States)

    Limon, Georgina; Gonzales-Gustavson, Eloy A; Gibson, Troy J

    2016-01-01

    Guinea pigs (Cavia porcelus) are an important source of nonhuman animal protein in the Andean region of South America. Specific guidelines regarding the welfare of guinea pigs before and during slaughter have yet to be developed. This study critically assessed the humaneness of 4 different stunning/slaughter methods for guinea pigs: cervical neck dislocation (n = 60), electrical head-only stunning (n = 83), carbon dioxide (CO2) stunning (n = 21), and penetrating captive bolt (n = 10). Following cervical neck dislocation, 97% of guinea pigs had at least 1 behavioral or cranial/spinal response. Six percent of guinea pigs were classified as mis-stunned after electrical stunning, and 1% were classified as mis-stunned after captive bolt. Increased respiratory effort was observed during CO2 stunning. Apart from this finding, there were no other obvious behavioral responses that could be associated with suffering. Of the methods assessed, captive bolt was deemed the most humane, effective, and practical method of stunning guinea pigs. Cervical neck dislocation should not be recommended as a slaughter method for guinea pigs.

  7. Temperature Preference in IAF Hairless and Hartley Guinea Pigs (Cavia porcellus).

    Science.gov (United States)

    Kleven, Gale A; Joshi, Prianca

    2016-03-01

    The Hairless strain of guinea pigs (Cavia porcellus) is the result of a spontaneous recessive mutation first identified at the Institute Armand Frappier (IAF) in 1978. Despite the longstanding availability of this strain, little is known about its thermoregulatory behavior. The aim of this study was to determine temperature preference in Hartley and Hairless guinea pigs by observing each strain in a ring-shaped apparatus containing a nonlinear temperature gradient. Temperatures were maintained by separately controlled heating mats lining the apparatus. Set point temperatures ranged from 24 to 38 °C. Guinea pigs (Hartley female, Hairless female, and Hairless male guinea pigs; n = 8 each group) were placed either singly or in pairs at 1 of the 8 randomized starting points within the apparatus. Subjects were observed for 30 min and coded for location within the temperature gradient by both frequency and duration. When placed singly in the apparatus, all 3 groups spent more time in the 30 °C zones. However, when placed as pairs with a cagemate, Hartley female guinea pigs spent more time in the cooler range of temperatures from 24 to 30 °C, whereas Hairless guinea pigs preferred a range of 30 to 38 °C. These results confirm a temperature preference of 30 ± 2 °C for both Hartley and Hairless guinea pigs when singly housed. However, data from the paired housing condition suggest that context plays an important role in thermoregulatory behavior.

  8. Congenital malformations caused by Stryphnodendron fissuratum (Leg. Mimosoideae) in guinea pigs (Cavia porcellus).

    Science.gov (United States)

    Macedo, Josenaldo S; Rocha, Brena P; Colodel, Edson M; Freitas, Sílvio H; Dória, Renata G S; Riet-Correa, Franklin; Evêncio-Neto, Joaquim; Mendonça, Fábio S

    2015-11-01

    The aim of this study was to evaluate the toxicity of Stryphnodendron fissuratum pods in guinea pigs (Cavia porcellus) and test the hypothesis that this plant has teratogenic effects. Thus, sixteen guinea pigs were randomly divided into four groups of four animals each. Groups 10, 20 and 40 consisted of guinea pigs that received commercial food that contained crushed pods of S. fissuratum at concentrations of 10, 20 and 40 g/kg, respectively, during the period of organogenesis. Control group consisted of guinea pigs under the same management conditions that did not receive crushed pods of S. fissuratum in their food. In all experimental groups, the main clinical signs of poisoning consisted of anorexia, prostration, absence of vocalizations, alopecia, diarrhea, and abortions within the adult guinea pigs. Those that did not abort gave birth to weak, malnourished pups, some of which had fetal malformations. The main teratogenic changes consisted of eventration, arthrogryposis, amelia of the forelimbs, anophthalmia, microphthalmia, anotia and agnathia. The reductions in the number of offspring and the malformations observed in the experimental groups suggest that S. fissuratum affects fetal development and is teratogenic.

  9. Detection of Aeromonas hydrophila Using Fiber Optic Microchannel Sensor

    Directory of Open Access Journals (Sweden)

    Samla Gauri

    2017-01-01

    Full Text Available This research focuses on the detection of Aeromonas hydrophila using fiber optic microchannel biosensor. Microchannel was fabricated by photolithography method. The fiber optic was chosen as signal transmitting medium and light absorption characteristic of different microorganisms was investigated for possible detection. Experimental results showed that Aeromonas hydrophila can be detected at the region of UV-Vis spectra between 352 nm and 354 nm which was comparable to measurement provided by UV spectrophotometer and also theoretical calculation by Beer-Lambert Absorption Law. The entire detection can be done in less than 10 minutes using a total volume of 3 μL only. This result promises good potential of this fiber optic microchannel sensor as a reliable, portable, and disposable sensor.

  10. Evolutionary Roots and Diversification of the Genus Aeromonas.

    Science.gov (United States)

    Sanglas, Ariadna; Albarral, Vicenta; Farfán, Maribel; Lorén, J G; Fusté, M C

    2017-01-01

    Despite the importance of diversification rates in the study of prokaryote evolution, they have not been quantitatively assessed for the majority of microorganism taxa. The investigation of evolutionary patterns in prokaryotes constitutes a challenge due to a very scarce fossil record, limited morphological differentiation and frequently complex taxonomic relationships, which make even species recognition difficult. Although the speciation models and speciation rates in eukaryotes have traditionally been established by analyzing the fossil record data, this is frequently incomplete, and not always available. More recently, several methods based on molecular sequence data have been developed to estimate speciation and extinction rates from phylogenies reconstructed from contemporary taxa. In this work, we determined the divergence time and temporal diversification of the genus Aeromonas by applying these methods widely used with eukaryotic taxa. Our analysis involved 150 Aeromonas strains using the concatenated sequences of two housekeeping genes (approximately 2,000 bp). Dating and diversification model analyses were performed using two different approaches: obtaining the consensus sequence from the concatenated sequences corresponding to all the strains belonging to the same species, or generating the species tree from multiple alignments of each gene. We used BEAST to perform a Bayesian analysis to estimate both the phylogeny and the divergence times. A global molecular clock cannot be assumed for any gene. From the chronograms obtained, we carried out a diversification analysis using several approaches. The results suggest that the genus Aeromonas began to diverge approximately 250 millions of years (Ma) ago. All methods used to determine Aeromonas diversification gave similar results, suggesting that the speciation process in this bacterial genus followed a rate-constant (Yule) diversification model, although there is a small probability that a slight

  11. Aeromonas spp.: evaluation of genomic diversity and biofilm forming ability

    OpenAIRE

    Craveiro, Sara Sofia Pereira, 1986-

    2013-01-01

    Tese de mestrado. Biologia (Microbiologia Aplicada). Universidade de Lisboa, Faculdade de Ciências, 2013 Aeromonas spp. are ubiquitous bacteria widely distributed among aquatic environments. Their persistence in water distribution systems is related to their ability to form biofilms, even in the presence of residual chlorine. Once in distribution water systems, aeromonads can contaminate drinking water, food processing facilities and food products. Moreover, members of this genus are known...

  12. Clinical and biochemical significance of toxin production by Aeromonas hydrophila.

    Science.gov (United States)

    Kindschuh, M; Pickering, L K; Cleary, T G; Ruiz-Palacios, G

    1987-05-01

    Production of cytotoxin and enterotoxin by Aeromonas strains obtained from stools of 50 children in Mexico and Texas and from blood of 9 children with sepsis was determined. Results were correlated with clinical features of infected children as well as with biochemical traits of Aeromonas strains. Cytotoxin was produced by 40 of 42 Aeromonas strains (95%) isolated from stools of children with diarrhea, by all 8 isolates from stools of well children, and by all 9 isolates from children with sepsis. There was no difference in the quantities (amount of cytotoxin per milligram of protein required to kill 50% of the cells) of cytotoxin produced and in clinical manifestations among the groups. None of the isolates produced a toxin that could be neutralized by antiserum raised against Shiga toxin produced by Shigella dysenteriae 1 60R. Heat-labile-like enterotoxin (LT) was produced by 26 of 42 stool isolates (62%), while only 1 of the 42 isolates (2%) produced enterotoxinlike activity in suckling mice; 65% of the cytotoxin-producing strains also produced an LT-like material. All strains from blood produced LT-like material, and 2 of 6 (33%) produced activity in suckling mice. All strains produced hemolysin; 37 of 57 (65%) were Voges-Proskauer positive; 27 of 57 (47%) were lysine decarboxylase positive by API 20E strips, none were positive for lysine decarboxylose production by lysin-iron agar slants at 24 h, but 17 of 54 (31%) were positive at 48 h. There was no correlation between biochemical reactions and enterotoxin or cytotoxin production. There appears to be no correlation between toxin production by Aeromonas spp. and gastroenteritis.

  13. Evolutionary Roots and Diversification of the Genus Aeromonas

    Science.gov (United States)

    Sanglas, Ariadna; Albarral, Vicenta; Farfán, Maribel; Lorén, J. G.; Fusté, M. C.

    2017-01-01

    Despite the importance of diversification rates in the study of prokaryote evolution, they have not been quantitatively assessed for the majority of microorganism taxa. The investigation of evolutionary patterns in prokaryotes constitutes a challenge due to a very scarce fossil record, limited morphological differentiation and frequently complex taxonomic relationships, which make even species recognition difficult. Although the speciation models and speciation rates in eukaryotes have traditionally been established by analyzing the fossil record data, this is frequently incomplete, and not always available. More recently, several methods based on molecular sequence data have been developed to estimate speciation and extinction rates from phylogenies reconstructed from contemporary taxa. In this work, we determined the divergence time and temporal diversification of the genus Aeromonas by applying these methods widely used with eukaryotic taxa. Our analysis involved 150 Aeromonas strains using the concatenated sequences of two housekeeping genes (approximately 2,000 bp). Dating and diversification model analyses were performed using two different approaches: obtaining the consensus sequence from the concatenated sequences corresponding to all the strains belonging to the same species, or generating the species tree from multiple alignments of each gene. We used BEAST to perform a Bayesian analysis to estimate both the phylogeny and the divergence times. A global molecular clock cannot be assumed for any gene. From the chronograms obtained, we carried out a diversification analysis using several approaches. The results suggest that the genus Aeromonas began to diverge approximately 250 millions of years (Ma) ago. All methods used to determine Aeromonas diversification gave similar results, suggesting that the speciation process in this bacterial genus followed a rate-constant (Yule) diversification model, although there is a small probability that a slight

  14. Screening and Characterization of a Bioflocculant Produced by Aeromonas sp.

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To isolate the bioflocculant-producing bacteria from activated sludge and investigate the flocculating characteristics of the newly isolated bioflocculant. Methods Bacteria were screened from activated sludge samples to isolate bioflocculant-producing bacteria. Flocculating activity was used as a measure of the flocculating capability of the bioflocculant.Results A novel bioflocculant-producing bacterium was isolated, which was identified to belong to genus Aeromonas and named as Aeromonas sp. N11. Flocculating activity increased in the presence of K+, Na+, or Ca2+. The highest flocculating activities for kaolin suspension were obtained in acidic pH ranges, and optimum pHs for it were 3.0, 4.0, and 5.0 with 1 mmol/L K+, Ca+, and Na+ present, respectively. The highest flocculating activities for soil suspension were observed at pH 8.0. The bioflocculant had a good flocculating activity and could achieve a flocculating activity of 92.4% for kaolin suspension at a Conclusion The bioflocculant produced by Aeromonas sp. N11 has strong flocculating activity and high stability, which affords high possibility of its practical use.

  15. Osteogenic and osteoclastogenic differentiation of co-cultured cells in polylactic acid-nanohydroxyapatite fiber scaffolds.

    Science.gov (United States)

    Morelli, Sabrina; Salerno, Simona; Holopainen, Jani; Ritala, Mikko; De Bartolo, Loredana

    2015-06-20

    The design of bone substitutes involves the creation of a microenvironment supporting molecular cross-talk between cells and scaffolds during tissue formation and remodelling. Bone remodelling process includes the cooperation of bone-building cells and bone-resorbing cells. In this paper we developed polylactic acid (PLA) and composite PLA-nanohydroxyapatite (nHA) scaffolds with 20 and 50wt.% of nHA by electrospinning technique to be used in bone tissue engineering. The developed scaffolds have different fiber diameter, porosity with interconnected pores and mechanical properties. Taking cues from the bone environment features we investigated the differentiation of human mesenchymal stem cells (hMSCs) from bone marrow in osteoblasts and the osteoclastogenesis in the developed scaffolds in homotypic and in co-culture up to 46 days. PLA and composite PLA-nHA scaffolds induced osteogenic and osteoclastogenic differentiation. Both osteoblasts and osteoclasts displayed high expression of specific markers (osteopontin, osteocalcin, RANK, RANKL) and functions such as secretion of ALP, cathepsin K and TRAP activity on composite scaffolds especially on PLA-nHA containing 20wt.% of nHA. The heterotypic interactions between osteoblasts and osteoclasts co-cultured in the developed scaffolds triggered their functional differentiation and activation.

  16. [A protocol for primary dissociated astrocyte and neuron co-culture].

    Science.gov (United States)

    Shi, Ying; Zhou, Mi; Jiang, Min

    2013-02-25

    Cultured primary hippocampal neurons are ideal tool for investigating the subcellular localization and trafficking of neuronal proteins. The aim of the present study was to establish a method to co-culture hippocampal neurons and cortical astrocytes, which would guarantee well conditions of neurons. Newborn Sprague-Dawley (SD) rats were sacrificed by decapitation. The cortex of cerebrum was cut into pieces, and the cortical tissue was digested with trypsin. The liquid suspension of single cells was planted onto a 25 cm² culture flask. On the fourth day of culture, the tissue cells except astrocytes were removed by intensive agitation of culture flask. Purified astrocytes were allowed to grow continuously until they reached most area of flask. At this time point, we replaced the culture media with neuronal cell media containing cytarabine, and planted the primary culture of rat hippocampal neurons onto the feed layer of cortical astrocytes. The microscopic observation results showed that, the astrocytes evenly grew without obvious boundaries between each other, and exhibited good purity. The co-cultured hippocampal neurons were in good condition, developed intertwined network of axons and dendrites, lived for a long time, and could tolerate gene transfection. Above all, this method is relatively simple from a technical point of view, yet provides healthy and reliable neuronal culture.

  17. Preliminary analysis of cellular sociology of co-cultured glioma initiating cells and macrophages in vitro

    Institute of Scientific and Technical Information of China (English)

    Mingxia Zhang; Xingliang Dai; Xiaonan Li; Qiang Huang; Jun Dong; Junjie Chen; Lin Wang; Xiaoyan Ji; Lin Yang; Yujing Sheng; Hairui Liu; Haiyang Wang; Aidong Wang

    2016-01-01

    Objective:Real-time monitoring of cytokine secretion at the single immunocyte level, based on the concept of immune cells, sociology has been recently reported. However, the relationships between glioma-initiating cells (GICs) and host immune cells and their mutual interactions in the tumor microenvironment have not been directly observed and remain unclear. Methods:The dual fluorescence tracing technique was applied to label the co-cultured GICs and host macrophages (Mø), and the interactions between the two types of cells were observed using a live cell imaging system. Fusion cells in the co-culture system were monocloned and proliferated in vitro and their social interactions were observed and recorded. Results:Using real-time dynamic observation of target cells, 6 types of intercellular conjunction microtubes were found to function in the transfer of intercellular information between GICs and Mø;GICs and host Mø can fuse into hybrid cells after several rounds of mutual interactions, and then these fusion cells fused with each other;Fusion cells generated offspring cells through symmetrical and asymmetrical division or underwent apoptosis. A“cell in cell” phenomenon was observed in the fusion cells, which was often followed by cell release, namely entosis. Conclusions:Preliminary studies revealed the patterns of cell conjunction via microtubes between GICs and host Mø and the processes of cell fusion, division, and entosis. The results revealed malignant transformation of host Mø, induced by GICs, suggesting complex social relationships among tumor-immune cells in gliomas.

  18. Functional genomic study of the environmentally important Desulfovibrio /Methanococcus syntrophic co-culture.

    Science.gov (United States)

    Mukhopadhyay, A.

    2008-12-01

    The use of microbe-oriented bioremediation for ameliorating extensive environmental pollution has fostered fundamental and applied studies of environmentally relevant microorganisms such as Desulfovibrio vulgaris, Shewanella oneidensis and Geobacter metallireducens.. Concurrently, there has been an increasing appreciation that the physiology of these organisms in pure culture is not necessarily representative of its activities in the environment. To enable a better understanding of microbial physiology under more environmentally relevant conditions, the syntrophic growth between the sulfate reducing bacterium, D. vulgaris and the hydrogenotrophic methanogen, Methanococcus maripaludis serves as an ideal system for laboratory studies. Cell wide analyses using transcript, proteomics and metabolite analysis have been widely used to understand cellular activity at a molecular level. Using D. vulgaris and M. maripaludis arrays, and the iTRAQ proteomics method, we studied the physiology of the D. vulgaris / M. maripaludis syntrophic co- cultures. The results from this study allowed us to identify differences in cellular response in mono-culture vs. co-culture growth for both D. vulgaris and M. maripaludis.

  19. Biomimetic injectable HUVEC-adipocytes/collagen/alginate microsphere co-cultures for adipose tissue engineering.

    Science.gov (United States)

    Yao, Rui; Zhang, Renji; Lin, Feng; Luan, Jie

    2013-05-01

    Engineering adipose tissue that has the ability to engraft and establish a vascular supply is a laudable goal that has broad clinical relevance, particularly for tissue reconstruction. In this article, we developed novel microtissues from surface-coated adipocyte/collagen/alginate microspheres and human umbilical vein endothelial cells (HUVECs) co-cultures that resembled the components and structure of natural adipose tissue. Firstly, collagen/alginate hydrogel microspheres embedded with viable adipocytes were obtained to mimic fat lobules. Secondly, collagen fibrils were allowed to self-assemble on the surface of the microspheres to mimic collagen fibrils surrounding the fat lobules in the natural adipose tissue and facilitate HUVEC attachment and co-cultures formation. Thirdly, the channels formed by the gap among the microspheres served as the room for in vitro prevascularization and in vivo blood vessel development. The endothelial cell layer outside the microspheres was a starting point of rapid vascular ingrowth. Adipose tissue formation was analyzed for 12 weeks at 4-week intervals by subcutaneous injection into the head of node mice. The vasculature in the regenerated tissue showed functional anastomosis with host blood vessels. Long-term stability of volume and weight of the injection was observed, indicating that the vasculature formed within the constructs benefited the formation, maturity, and maintenance of adipose tissue. This study provides a microsurgical method for adipose regeneration and construction of biomimetic model for drug screening studies.

  20. Developing a thermophilic hydrogen-producing co-culture for efficient utilization of mixed sugars

    Energy Technology Data Exchange (ETDEWEB)

    Zeidan, Ahmad A.; Van Niel, Ed W.J. [Department of Applied Microbiology, Lund University, P.O. Box 124, SE-221 00 Lund (Sweden)

    2009-05-15

    Previous studies on the extreme thermophile Caldicellulosiruptor saccharolyticus revealed that the organism produces high yields of hydrogen on glucose and xylose, the major components of lignocellulosic hydrolysates. Preliminary experiments on mixed sugar substrates, however, indicated that xylose was preferred over glucose. The sugar preference of some other extreme thermophiles, including Caldicellulosiruptor owensensis, Caldicellulosiruptor kristjanssonii and newly enriched, thermophilic compost sludge microflora, was investigated in an attempt to find complementary organisms to C. saccharolyticus for rapid and efficient utilization of lignocellulosic sugars. The behavior of C. owensensis and C. kristjanssonii appeared to be similar to that of C. saccharolyticus, either in pure cultures or in co-cultures with the latter. Co-culturing C. saccharolyticus with the enriched compost microflora resulted in fast, simultaneous consumption of both glucose and xylose in the medium with a relatively high specific hydrogen production rate, 40 mmol (gCDW){sup -1} h{sup -1}, and high volumetric productivity, 22.5 mmol l{sup -1} h{sup -1}. (author)

  1. Production of angkak through co-culture of Monascus purpureus and Monascus ruber

    Directory of Open Access Journals (Sweden)

    Bibhu Prasad Panda

    2010-10-01

    Full Text Available Angkak (red mold rice, red yeast rice, Chinese red rice is a traditional Chinese medicine produced by solid-state fermentation of cooked non-glutinous rice with Monascus species. The secondary metabolite of Monascus species, monacolin K /lovastatin, has been proven to lower blood lipid levels. In this study, a co-culture of Monascus purpureus MTCC 369 and Monascus ruber MTCC 1880 was used for angkak production. Four medium parameters screened by Plackett-Burman design were optimized by response surface methodology for highest lovastatin production in angkak during solid-state fermentation by the co-culture. Maximum lovastatin production of 2.84 mg g-1 was predicted in solid medium containing 20 g rice and 40 ml liquid nutrients medium (malt extract 9.68 g l-1, dextrose 38.90 g l-1, MnSO4.H2O 1.96 g l-1, and MgSO4.7H2O 0.730 g l-1 by point prediction tool of Design Expert 7.1 software (Statease Inc. USA.

  2. Comparative genomic study of gastric epithelial cells co-cultured with Helicobacter pylori

    Institute of Scientific and Technical Information of China (English)

    Fen Wang; Li-Dan Luo; Jian-Hua Pan; Li-Hua Huang; Hong-Wei Lv; Qin Guo; Can-Xia Xu

    2012-01-01

    AIM:To identify genes potentially involved in Helicobacter pylori (H.pylori)-induced gastric carcinogenesis.METHODS:GES-1 cells were co-cultured with H.pylori strains isolated from patients with gastric carcinoma (GC,n =10) or chronic gastritis (CG,n =10) for in vitro proliferation and apoptosis assays to identify the most and least virulent strains.These two strains were cagA-genotyped and used for further in vivo carcinogenic virulence assays by infecting Mongolian gerbils for 52 wk,respectively; a broth free of H.pylori was lavaged as control.Genomic profiles of GES-1 cells cocultured with the most and least virulent strains were determined by microarray analysis.The most differentially expressed genes were further verified using quantitative real-time polymerase chain reaction in GES-1 cells infected with the most and least virulent strains,and by immunohistochemistry in H.pylori positive CG,precancerous diseases,and GC biopsy specimens in an independent experiment.RESULTS:GC-derived H.pylori strains induced a potent proliferative effect in GES-1 cells in co-culture,whereas CG-derived strains did not.The most (from a GC patient) and least (from a CG patient) virulent strains were cagA-positive and negative,respectively.At week 52,CG,atrophy,metaplasia,dysplasia,and GC were observed in 90.0%,80.0%,80.0%,90%,and 60.0%,respectively,of the animals lavaged with the most virulent strain.However,only mild CG was observed in 90% of the animals lavaged with the least virulent strain.On microarray analysis,800 differentially expressed genes (49 up-and 751 down-regulated),involving those associated with cell cycle regulation,cell apoptosis,cytoskeleton,immune response,and substance and energy metabolisms,were identified in cells co-cultured with the most virulent strain as compared with those co-cultured with the least virulent strain.The six most differentially expressed genes (with a betweenness centrality of 0.1-0.2) were identified among the significant

  3. Comparison of fibre digestion and digesta retention time between nutrias (Myocaster coypus) and guinea-pigs (Cavia porcellus).

    Science.gov (United States)

    Sakaguchi, E; Nabata, A

    1992-11-01

    1. Digestibilities of feed, and transit and retention time of fluid and particle digesta marker measured in nutrias (Myocaster coypus) and guinea-pigs (Cavia porcellus) fed on a diet containing 50% alfalfa. 2. The digestibility of fibre was higher in the nutria, along with the longer retention time of digesta. 3. The liquid and particle marker were similarly excreted, suggesting no separation mechanism in the gastrointestinal tract of both the animals. 4. The apparent digestibility of protein in the nutria was superior to the guinea-pig and other small hindgut fermenters, suggesting that the contribution of coprophagy on protein nutrition of nutrias is significant.

  4. EVALUACIÓN DE DOS GENOTIPOS DE CUYES (Cavia porcellus) ALIMENTADOS CON CONCENTRADO Y EXCLUSIÓN DE FORRAJE VERDE.

    OpenAIRE

    Camino M., Javier; Departamento de Nutrición, Facultad de Zootecnia, Universidad Nacional Agraria La Molina, Lima.; Hidalgo L., Víctor; Departamento de Nutrición, Facultad de Zootecnia, Universidad Nacional Agraria La Molina, Lima.

    2014-01-01

    El estudio tuvo como objetivo evaluar los parámetros productivos y el porcentaje de grasa en la carcasa de dos genotipos de cuyes (Cavia porcellus) alimentados por nueve semanas con dos tipos de dieta. Se utilizó el diseño estadístico completamente al azar con arreglo factorial 2x2, teniendo como factores dos genotipos (Cieneguilla-UNALM y Perú-INIA) y dos tipos de dietas (dieta 1: alimento balanceado, forraje verde y agua; dieta 2: alimento balanceado más vitamina C y agua), y se evaluó el p...

  5. Diarrea del viajero asociada a Aeromonas hydrophila Traveller's diarrohea caused by Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    M. L. Pita Toledo

    2001-06-01

    Full Text Available Aeromonas hydrophila es una bacteria que se encuentra frecuentemente en aguas dulces y salobres. Desde hace varios años se conoce que determinadas cepas de la especie pueden causar enfermedad en pescados y en anfibios, existiendo cierta controversia sobre su patogeneicidad en humanos. No obstante, puede dar lugar a infección en personas, que la adquieren bien a través de heridas abiertas o bien por ingestión de alimentos o agua contaminados. Sin embargo, en prácticamente la totalidad de las publicaciones se hace referencia a casos aislados más que a brotes producidos por esta bacteria. Generalmente la infección por A. hydrophila da lugar a gastroenteritis en personas sanas, pudiendo generar una septicemia en individuos inmunodeprimidos.En 1998 se produjo una alarma en el sector hotelero por un incremento considerable de los casos de gastroenteritis entre los turistas que acudían durante sus vacaciones a determinadas zonas costeras de la isla de Tenerife.Tras tener conocimiento de la situación, la Dirección General de Salud Pública inició una investigación epidemiológica con el objetivo de conocer la magnitud real del problema detectado e identificar los factores asociados a la misma, para poner en marcha las medidas correctoras y de control necesarias. El análisis inicial permitió determinar que se trataba de un proceso de gran magnitud en su extensión aunque de carácter leve, que afectaba únicamente a los turistas en una zona costera en concreto, compatible por lo tanto con la diarrea del viajero, planteándose un origen probablemente hídrico.Aeromonas hydrophila is a bacteria frequently found in fresh water and saline water. Certain strains of the species have been known to cause disease in fish and amphibians for several years, although some controversy exists about its ability to cause illness in humans. Nevertheless, either through open wounds or contaminated food or water, it can give rise to infection. However, almost

  6. Aeromonas hydrophila urinary tract infection in pregnancy- Case report and literature review

    Directory of Open Access Journals (Sweden)

    Latha Ragunathan

    2012-03-01

    Full Text Available A case of a pregnant woman without previous or concomitant disease, who developed an Aeromonas hydrophila urinarytract infection (UTI at 12 weeks gestation, is reported. A brief review of the literature on the association and incidenceof Aeromonas spp in urinary infections and also in association with pregnancy is presented. J Microbiol Infect Dis 2012;2(1: 26-28

  7. Chitin degradation and utilization by virulent Aeromonas hydrophila strain ML10-51K

    Science.gov (United States)

    Virulent Aeromonas hydrophila (vAh) is one of the most important bacterial pathogens that causes persistent outbreaks of motile Aeromonas septicemia (MAS) in warm-water fishes. Among factors associated with MAS outbreaks, the survivability of this pathogen in aquatic environments is of great concern...

  8. Complete genome sequence of channel catfish epidemic isolate Aeromonas hydrophila ML09-119

    Science.gov (United States)

    Aeromonas hydrophila is a Gram-negative, rod-shaped, mesophilic bacteria that infects both aquatic poikilothermic animals and mammals, including humans. Here, we present the complete genome sequence of Aeromonas hydrophila ML-09-119, which represents a clonal group of A. hydrophila isolates causing ...

  9. Classification of a hypervirulent Aeromonas hydrophila pathotype responsible for epidemic outbreaks in warm-water fishes

    Science.gov (United States)

    Lineages of hypervirulent Aeromonas hydrophila (vAh) are the cause of persistent outbreaks of motile Aeromonas septicemia in warm-water fishes worldwide. Over the last decade, this virulent lineage of A. hydrophila has resulted in annual losses of millions of tons of farmed carp and catfish in the P...

  10. Fermentative hydrogen production from glucose and starch using pure strains and artificial co-cultures ofClostridium spp.

    Directory of Open Access Journals (Sweden)

    Masset Julien

    2012-05-01

    Full Text Available Abstract Background Pure bacterial strains give better yields when producing H2 than mixed, natural communities. However the main drawback with the pure cultures is the need to perform the fermentations under sterile conditions. Therefore, H2 production using artificial co-cultures, composed of well characterized strains, is one of the directions currently undertaken in the field of biohydrogen research. Results Four pure Clostridium cultures, including C. butyricum CWBI1009, C. pasteurianum DSM525, C. beijerinckii DSM1820 and C. felsineum DSM749, and three different co-cultures composed of (1 C. pasteurianum and C. felsineum, (2 C. butyricum and C. felsineum, (3 C. butyricum and C. pasteurianum, were grown in 20 L batch bioreactors. In the first part of the study a strategy composed of three-culture sequences was developed to determine the optimal pH for H2 production (sequence 1; and the H2-producing potential of each pure strain and co-culture, during glucose (sequence 2 and starch (sequence 3 fermentations at the optimal pH. The best H2 yields were obtained for starch fermentations, and the highest yield of 2.91 mol H2/ mol hexose was reported for C. butyricum. By contrast, the biogas production rates were higher for glucose fermentations and the highest value of 1.5 L biogas/ h was observed for the co-culture (1. In general co-cultures produced H2 at higher rates than the pure Clostridium cultures, without negatively affecting the H2 yields. Interestingly, all the Clostridium strains and co-cultures were shown to utilize lactate (present in a starch-containing medium, and C. beijerinckii was able to re-consume formate producing additional H2. In the second part of the study the co-culture (3 was used to produce H2 during 13 days of glucose fermentation in a sequencing batch reactor (SBR. In addition, the species dynamics, as monitored by qPCR (quantitative real-time PCR, showed a stable coexistence of C. pasteurianum and C

  11. Discovery of novel xylosides in co-culture of basidiomycetes Trametes versicolor and Ganoderma applanatum by integrated metabolomics and bioinformatics

    Science.gov (United States)

    Yao, Lu; Zhu, Li-Ping; Xu, Xiao-Yan; Tan, Ling-Ling; Sadilek, Martin; Fan, Huan; Hu, Bo; Shen, Xiao-Ting; Yang, Jie; Qiao, Bin; Yang, Song

    2016-01-01

    Transcriptomic analysis of cultured fungi suggests that many genes for secondary metabolite synthesis are presumably silent under standard laboratory condition. In order to investigate the expression of silent genes in symbiotic systems, 136 fungi-fungi symbiotic systems were built up by co-culturing seventeen basidiomycetes, among which the co-culture of Trametes versicolor and Ganoderma applanatum demonstrated the strongest coloration of confrontation zones. Metabolomics study of this co-culture discovered that sixty-two features were either newly synthesized or highly produced in the co-culture compared with individual cultures. Molecular network analysis highlighted a subnetwork including two novel xylosides (compounds 2 and 3). Compound 2 was further identified as N-(4-methoxyphenyl)formamide 2-O-β-D-xyloside and was revealed to have the potential to enhance the cell viability of human immortalized bronchial epithelial cell line of Beas-2B. Moreover, bioinformatics and transcriptional analysis of T. versicolor revealed a potential candidate gene (GI: 636605689) encoding xylosyltransferases for xylosylation. Additionally, 3-phenyllactic acid and orsellinic acid were detected for the first time in G. applanatum, which may be ascribed to response against T.versicolor stress. In general, the described co-culture platform provides a powerful tool to discover novel metabolites and help gain insights into the mechanism of silent gene activation in fungal defense. PMID:27616058

  12. β-Carotene from Yeasts Enhances Laccase Production of Pleurotus eryngii var. ferulae in Co-culture

    Directory of Open Access Journals (Sweden)

    Chaolin Guo

    2017-06-01

    Full Text Available Laccase is widely used in several industrial applications and co-culture is a common method for enhancing laccase production in submerged fermentation. In this study, the co-culture of four yeasts with Pleurotus eryngii var. ferulae was found to enhance laccase production. An analysis of sterilization temperatures and extraction conditions revealed that the stimulatory compound in yeasts was temperature-sensitive, and that it was fat-soluble. An LC-MS analysis revealed that the possible stimulatory compound for laccase production in the four yeast extracts was β-carotene. Moreover, the addition of 4 mg β-carotene to 150 mL of P. eryngii var. ferulae culture broth improved laccase production by 2.2-fold compared with the control (i.e., a monoculture, and was similar to laccase production in co-culture. In addition, the enhanced laccase production was accompanied by an increase of lac gene transcription, which was 6.2-time higher than the control on the fifth day. Therefore, it was concluded that β-carotene from the co-cultured yeasts enhanced laccase production in P. eryngii var. ferulae, and strains that produce β-carotene could be selected to enhance fungal laccase production in a co-culture. Alternatively, β-carotene or crude extracts of β-carotene could be used to induce high laccase production in large scale.

  13. Expanding the Diet for DIET: Electron Donors Supporting Direct Interspecies Electron Transfer (DIET in Defined Co-Cultures

    Directory of Open Access Journals (Sweden)

    Li-YIng eWang

    2016-03-01

    Full Text Available Direct interspecies electron transfer (DIET has been recognized as an alternative to interspecies H2 transfer as a mechanism for syntrophic growth, but previous studies on DIET with defined co-cultures have only documented DIET with ethanol as the electron donor in the absence of conductive materials. Co-cultures of Geobacter metallireducens and Geobacter sulfurreducens metabolized propanol, butanol, propionate, and butyrate with the reduction of fumarate to succinate. G. metallireducens utilized each of these substrates whereas only electrons available from DIET supported G. sulfurreducens respiration. A co-culture of G. metallireducens and a strain of G. sulfurreducens that could not metabolize acetate oxidized acetate with fumarate as the electron acceptor, demonstrating that acetate can also be syntrophically metabolized via DIET. A co-culture of G. metallireducens and Methanosaeta harundinacea previously shown to syntrophically convert ethanol to methane via DIET metabolized propanol or butanol as the sole electron donor, but not propionate or butyrate. The stoichiometric accumulation of propionate or butyrate in the propanol- or butanol-fed cultures demonstrated that M. harundinaceae could conserve energy to support growth solely from electrons derived from DIET. Co-cultures of G. metallireducens and Methanosarcina barkeri could also incompletely metabolize propanol and butanol and did not metabolize propionate or butyrate as sole electron donors. These results expand the range of substrates that are known to be syntrophically metabolized through DIET, but suggest that claims of propionate and butyrate metabolism via DIET in mixed microbial communities warrant further validation.

  14. Experimental and computational optimization of an Escherichia coli co-culture for the efficient production of flavonoids.

    Science.gov (United States)

    Jones, J Andrew; Vernacchio, Victoria R; Sinkoe, Andrew L; Collins, Shannon M; Ibrahim, Mohammad H A; Lachance, Daniel M; Hahn, Juergen; Koffas, Mattheos A G

    2016-05-01

    Metabolic engineering and synthetic biology have enabled the use of microbial production platforms for the renewable production of many high-value natural products. Titers and yields, however, are often too low to result in commercially viable processes. Microbial co-cultures have the ability to distribute metabolic burden and allow for modular specific optimization in a way that is not possible through traditional monoculture fermentation methods. Here, we present an Escherichia coli co-culture for the efficient production of flavonoids in vivo, resulting in a 970-fold improvement in titer of flavan-3-ols over previously published monoculture production. To accomplish this improvement in titer, factors such as strain compatibility, carbon source, temperature, induction point, and inoculation ratio were initially optimized. The development of an empirical scaled-Gaussian model based on the initial optimization data was then implemented to predict the optimum point for the system. Experimental verification of the model predictions resulted in a 65% improvement in titer, to 40.7±0.1mg/L flavan-3-ols, over the previous optimum. Overall, this study demonstrates the first application of the co-culture production of flavonoids, the most in-depth co-culture optimization to date, and the first application of empirical systems modeling for improvement of titers from a co-culture system.

  15. Discovery of novel xylosides in co-culture of basidiomycetes Trametes versicolor and Ganoderma applanatum by integrated metabolomics and bioinformatics

    Science.gov (United States)

    Yao, Lu; Zhu, Li-Ping; Xu, Xiao-Yan; Tan, Ling-Ling; Sadilek, Martin; Fan, Huan; Hu, Bo; Shen, Xiao-Ting; Yang, Jie; Qiao, Bin; Yang, Song

    2016-09-01

    Transcriptomic analysis of cultured fungi suggests that many genes for secondary metabolite synthesis are presumably silent under standard laboratory condition. In order to investigate the expression of silent genes in symbiotic systems, 136 fungi-fungi symbiotic systems were built up by co-culturing seventeen basidiomycetes, among which the co-culture of Trametes versicolor and Ganoderma applanatum demonstrated the strongest coloration of confrontation zones. Metabolomics study of this co-culture discovered that sixty-two features were either newly synthesized or highly produced in the co-culture compared with individual cultures. Molecular network analysis highlighted a subnetwork including two novel xylosides (compounds 2 and 3). Compound 2 was further identified as N-(4-methoxyphenyl)formamide 2-O-β-D-xyloside and was revealed to have the potential to enhance the cell viability of human immortalized bronchial epithelial cell line of Beas-2B. Moreover, bioinformatics and transcriptional analysis of T. versicolor revealed a potential candidate gene (GI: 636605689) encoding xylosyltransferases for xylosylation. Additionally, 3-phenyllactic acid and orsellinic acid were detected for the first time in G. applanatum, which may be ascribed to response against T.versicolor stress. In general, the described co-culture platform provides a powerful tool to discover novel metabolites and help gain insights into the mechanism of silent gene activation in fungal defense.

  16. Supportive angiogenic and osteogenic differentiation of mesenchymal stromal cells and endothelial cells in monolayer and co-cultures

    Institute of Scientific and Technical Information of China (English)

    Florian Böhrnsen; Henning Schliephake

    2016-01-01

    Sites of implantation with compromised biology may be unable to achieve the required level of angiogenic and osteogenic regeneration. The specific function and contribution of different cell types to the formation of prevascularized, osteogenic networks in co-culture remains unclear. To determine how bone marrow-derived mesenchymal stromal cells (BMSCs) and endothelial cells (ECs) contribute to cellular proangiogenic differentiation, we analysed the differentiation of BMSCs and ECs in standardized monolayer, Transwell and co-cultures. BMSCs were derived from the iliac bone marrow of five patients, characterized and differentiated in standardized monolayers, permeable Transwells and co-cultures with human umbilical vein ECs (HUVECs). The expression levels of CD31, von Willebrand factor, osteonectin (ON) and Runx2 were assessed by quantitative reverse transcriptase polymerase chain reaction. The protein expression of alkaline phosphatase, ON and CD31 was demonstrated via histochemical and immunofluorescence analysis. The results showed that BMSCs and HUVECs were able to retain their lineage-specific osteogenic and angiogenic differentiation in direct and indirect co-cultures. In addition, BMSCs demonstrated a supportive expression of angiogenic function in co-culture, while HUVEC was able to improve the expression of osteogenic marker molecules in BMSCs.

  17. Bilaminar co-culture of primary rat cortical neurons and glia.

    Science.gov (United States)

    Shimizu, Saori; Abt, Anna; Meucci, Olimpia

    2011-11-12

    This video will guide you through the process of culturing rat cortical neurons in the presence of a glial feeder layer, a system known as a bilaminar or co-culture model. This system is suitable for a variety of experimental needs requiring either a glass or plastic growth substrate and can also be used for culture of other types of neurons. Rat cortical neurons obtained from the late embryonic stage (E17) are plated on glass coverslips or tissue culture dishes facing a feeder layer of glia grown on dishes or plastic coverslips (known as Thermanox), respectively. The choice between the two configurations depends on the specific experimental technique used, which may require, or not, that neurons are grown on glass (e.g. calcium imaging versus Western blot). The glial feeder layer, an astroglia-enriched secondary culture of mixed glia, is separately prepared from the cortices of newborn rat pups (P2-4) prior to the neuronal dissection. A major advantage of this culture system as compared to a culture of neurons only is the support of neuronal growth, survival, and differentiation provided by trophic factors secreted from the glial feeder layer, which more accurately resembles the brain environment in vivo. Furthermore, the co-culture can be used to study neuronal-glial interactions(1). At the same time, glia contamination in the neuronal layer is prevented by different means (low density culture, addition of mitotic inhibitors, lack of serum and use of optimized culture medium) leading to a virtually pure neuronal layer, comparable to other established methods(1-3). Neurons can be easily separated from the glial layer at any time during culture and used for different experimental applications ranging from electrophysiology(4), cellular and molecular biology(5-8), biochemistry(5), imaging and microscopy(4,6,7,9,10). The primary neurons extend axons and dendrites to form functional synapses(11), a process which is not observed in neuronal cell lines, although some

  18. Aeromonas detection and characterization using genus-specific PCR and single-strand conformation polymorphism (SSCP).

    Science.gov (United States)

    Delamare, Ana Paula Longaray; Lucena, Roberto Francisco; Thomazi, Guilherme; Ferrarini, Shana; Zacaria, Jucimar; Echeverrigaray, Sergio

    2012-10-01

    Based on sequence alignment, oligonucleotide primers targeting the Aeromonas extracellular lipase gene were developed for PCR detection of member of the genus. A pair of primers designed for conserved regions of the gene amplified a 276 bp sequence in all Aeromonas species and tested strains, but did not have a positive result with other Gram-positive and Gram-negative bacteria, showing high specificity and sensitivity. Selective enrichment in alkaline peptone water, followed by centrifugation, and direct usage of cells suspension as template, detected initial populations of 10 c.f.u. ml⁻¹. Single-strand conformation polymorphism analysis of the PCR products allowed the characterization of Aeromonas strains with a high discriminatory power (Simpson's index = 0.988). The method presented here provides a useful tool for the rapid detection of Aeromonas and the characterization of Aeromonas isolates.

  19. A First Generation Comparative Chromosome Map between Guinea Pig (Cavia porcellus) and Humans.

    Science.gov (United States)

    Romanenko, Svetlana A; Perelman, Polina L; Trifonov, Vladimir A; Serdyukova, Natalia A; Li, Tangliang; Fu, Beiyuan; O'Brien, Patricia C M; Ng, Bee L; Nie, Wenhui; Liehr, Thomas; Stanyon, Roscoe; Graphodatsky, Alexander S; Yang, Fengtang

    2015-01-01

    The domesticated guinea pig, Cavia porcellus (Hystricomorpha, Rodentia), is an important laboratory species and a model for a number of human diseases. Nevertheless, genomic tools for this species are lacking; even its karyotype is poorly characterized. The guinea pig belongs to Hystricomorpha, a widespread and important group of rodents; so far the chromosomes of guinea pigs have not been compared with that of other hystricomorph species or with any other mammals. We generated full sets of chromosome-specific painting probes for the guinea pig by flow sorting and microdissection, and for the first time, mapped the chromosomal homologies between guinea pig and human by reciprocal chromosome painting. Our data demonstrate that the guinea pig karyotype has undergone extensive rearrangements: 78 synteny-conserved human autosomal segments were delimited in the guinea pig genome. The high rate of genome evolution in the guinea pig may explain why the HSA7/16 and HSA16/19 associations presumed ancestral for eutherians and the three syntenic associations (HSA1/10, 3/19, and 9/11) considered ancestral for rodents were not found in C. porcellus. The comparative chromosome map presented here is a starting point for further development of physical and genetic maps of the guinea pig as well as an aid for genome assembly assignment to specific chromosomes. Furthermore, the comparative mapping will allow a transfer of gene map data from other species. The probes developed here provide a genomic toolkit, which will make the guinea pig a key species to unravel the evolutionary biology of the Hystricomorph rodents.

  20. Female novelty and the courtship behavior of male guinea pigs (Cavia porcellus).

    Science.gov (United States)

    Cohn, D W H; Tokumaru, R S; Ades, C

    2004-06-01

    In several rodent species, an increase or recovery of sexual behavior can be observed when sexually satiated males are placed in contact with a novel mate. In order to assess the influence of female novelty on the courtship behavior of guinea pigs (Cavia porcellus), four adult males were observed during four daily 15-min sessions while interacting with the same pregnant female (same-female sessions). A new female was presented during the fifth session (switched-female session). The duration of behavioral categories was obtained from videotape records using an observational software. From the first to the second session, all males decreased the time allocated to investigating (sniffing and licking), following, and mounting the female, and that response did not recover by the end of the same-female sessions. No similar decreasing tendencies were detected in the circling or rumba categories. A marked increase of investigating occurred in all males from the last same-female session (8.1, 11.9, 15.1 and 17.3 percent session time) to the switched-female one (16.4, 18.4, 37.1 and 28.9 percent session time, respectively). Increases in following and circling were recorded in three of four males, and full-blown recovery of mounting in one male. No consistent changes in the females' responses to males (following or attacking) were observed throughout testing. These results are consistent with the hypothesis that guinea pig males recognize individual females and that courtship responses may suffer a habituation/recovery process controlled by mate novelty.

  1. Female novelty and the courtship behavior of male guinea pigs (Cavia porcellus

    Directory of Open Access Journals (Sweden)

    Cohn D.W.H.

    2004-01-01

    Full Text Available In several rodent species, an increase or recovery of sexual behavior can be observed when sexually satiated males are placed in contact with a novel mate. In order to assess the influence of female novelty on the courtship behavior of guinea pigs (Cavia porcellus, four adult males were observed during four daily 15-min sessions while interacting with the same pregnant female (same-female sessions. A new female was presented during the fifth session (switched-female session. The duration of behavioral categories was obtained from videotape records using an observational software. From the first to the second session, all males decreased the time allocated to investigating (sniffing and licking, following, and mounting the female, and that response did not recover by the end of the same-female sessions. No similar decreasing tendencies were detected in the circling or rumba categories. A marked increase of investigating occurred in all males from the last same-female session (8.1, 11.9, 15.1 and 17.3 percent session time to the switched-female one (16.4, 18.4, 37.1 and 28.9 percent session time, respectively. Increases in following and circling were recorded in three of four males, and full-blown recovery of mounting in one male. No consistent changes in the females' responses to males (following or attacking were observed throughout testing. These results are consistent with the hypothesis that guinea pig males recognize individual females and that courtship responses may suffer a habituation/recovery process controlled by mate novelty.

  2. Validation of a Behavioral Ethogram for Assessing Postoperative Pain in Guinea Pigs (Cavia porcellus).

    Science.gov (United States)

    Dunbar, Misha L; David, Emily M; Aline, Marian R; Lofgren, Jennifer L

    2016-01-01

    Although guinea pigs (Cavia porcellus) have been used in research for more than a century and remain the most prevalent USDA-covered species, little has been elucidated regarding the recognition of clinical pain or analgesic efficacy in this species. We sought to assess pain in guinea pigs by using newer, clinically relevant methods that have been validated in other rodent species: the behavioral ethogram and cageside proxy indicator. In this study, 10 male guinea pigs underwent electronic von Frey testing of nociception, remote videorecording of behavior, and cageside assessment by using time-to-consumption (TTC) of a preferred treat test. These assessments were performed across 2 conditions (anesthesia only and castration surgery under anesthesia) at 3 time points (2, 8, and 24 h after the event). The anesthesia only condition served to control for the nonpainful but potentially distressing components of the surgical experience. Compared with those after anesthesia only conditions, subtle body movements were increased and nociceptive thresholds were decreased at 2 and 8 h after surgery. At 24 h, neither subtle body movement behaviors nor nociceptive thresholds differed between the 2 conditions. In contrast, TTC scores did not differ between the anesthesia only and surgery conditions at any time point, underscoring the challenge of identifying pain in this species through cageside evaluation. By comparing ethogram scores with measures of nociception, we validated select behaviors as pain-specific. Therefore, our novel ethogram allowed us to assess postoperative pain and may further serve as a platform for future analgesia efficacy studies in guinea pigs.

  3. Trixacarus caviae Fain, Howell & Hyatt 1972 (Acari: Sarcoptidae) as a cause of mange in guinea-pigs and papular urticaria in man

    NARCIS (Netherlands)

    Dorrestein, G.M.; Bronswijk, J.E.M.H. van

    1979-01-01

    The clinical symptoms of infestation with Trixacarus caviae in guinea-pigs are described. In general large areas of thickened, denuded skin, with a heavy secondary bacterial infection, and sometimes neurological signs, are observed in mangy cases. Antiparasitic therapy was successful with one or sev

  4. Trixacarus caviae Fain, Howell & Hyatt 1972 (Acari: Sarcoptidae) as a cause of mange in guinea-pigs and papular urticaria in man

    NARCIS (Netherlands)

    Dorrestein, G.M.; Bronswijk, J.E.M.H. van

    1979-01-01

    The clinical symptoms of infestation with Trixacarus caviae in guinea-pigs are described. In general large areas of thickened, denuded skin, with a heavy secondary bacterial infection, and sometimes neurological signs, are observed in mangy cases. Antiparasitic therapy was successful with one or sev

  5. Generalized Additive Mixed-Models for Pharmacology Using Integrated Discrete Multiple Organ Co-Culture

    Science.gov (United States)

    Ingersoll, Thomas; Cole, Stephanie; Madren-Whalley, Janna; Booker, Lamont; Dorsey, Russell; Li, Albert; Salem, Harry

    2016-01-01

    Integrated Discrete Multiple Organ Co-culture (IDMOC) is emerging as an in-vitro alternative to in-vivo animal models for pharmacology studies. IDMOC allows dose-response relationships to be investigated at the tissue and organoid levels, yet, these relationships often exhibit responses that are far more complex than the binary responses often measured in whole animals. To accommodate departure from binary endpoints, IDMOC requires an expansion of analytic techniques beyond simple linear probit and logistic models familiar in toxicology. IDMOC dose-responses may be measured at continuous scales, exhibit significant non-linearity such as local maxima or minima, and may include non-independent measures. Generalized additive mixed-modeling (GAMM) provides an alternative description of dose-response that relaxes assumptions of independence and linearity. We compared GAMMs to traditional linear models for describing dose-response in IDMOC pharmacology studies. PMID:27110941

  6. In Vitro Co-Culture Models of Breast Cancer Metastatic Progression towards Bone

    Directory of Open Access Journals (Sweden)

    Chiara Arrigoni

    2016-08-01

    Full Text Available Advanced breast cancer frequently metastasizes to bone through a multistep process involving the detachment of cells from the primary tumor, their intravasation into the bloodstream, adhesion to the endothelium and extravasation into the bone, culminating with the establishment of a vicious cycle causing extensive bone lysis. In recent years, the crosstalk between tumor cells and secondary organs microenvironment is gaining much attention, being indicated as a crucial aspect in all metastatic steps. To investigate the complex interrelation between the tumor and the microenvironment, both in vitro and in vivo models have been exploited. In vitro models have some advantages over in vivo, mainly the possibility to thoroughly dissect in controlled conditions and with only human cells the cellular and molecular mechanisms underlying the metastatic progression. In this article we will review the main results deriving from in vitro co-culture models, describing mechanisms activated in the crosstalk between breast cancer and bone cells which drive the different metastatic steps.

  7. In Vitro Co-Culture Models of Breast Cancer Metastatic Progression towards Bone

    Science.gov (United States)

    Arrigoni, Chiara; Bersini, Simone; Gilardi, Mara; Moretti, Matteo

    2016-01-01

    Advanced breast cancer frequently metastasizes to bone through a multistep process involving the detachment of cells from the primary tumor, their intravasation into the bloodstream, adhesion to the endothelium and extravasation into the bone, culminating with the establishment of a vicious cycle causing extensive bone lysis. In recent years, the crosstalk between tumor cells and secondary organs microenvironment is gaining much attention, being indicated as a crucial aspect in all metastatic steps. To investigate the complex interrelation between the tumor and the microenvironment, both in vitro and in vivo models have been exploited. In vitro models have some advantages over in vivo, mainly the possibility to thoroughly dissect in controlled conditions and with only human cells the cellular and molecular mechanisms underlying the metastatic progression. In this article we will review the main results deriving from in vitro co-culture models, describing mechanisms activated in the crosstalk between breast cancer and bone cells which drive the different metastatic steps. PMID:27571063

  8. Generalized Additive Mixed-Models for Pharmacology Using Integrated Discrete Multiple Organ Co-Culture.

    Directory of Open Access Journals (Sweden)

    Thomas Ingersoll

    Full Text Available Integrated Discrete Multiple Organ Co-culture (IDMOC is emerging as an in-vitro alternative to in-vivo animal models for pharmacology studies. IDMOC allows dose-response relationships to be investigated at the tissue and organoid levels, yet, these relationships often exhibit responses that are far more complex than the binary responses often measured in whole animals. To accommodate departure from binary endpoints, IDMOC requires an expansion of analytic techniques beyond simple linear probit and logistic models familiar in toxicology. IDMOC dose-responses may be measured at continuous scales, exhibit significant non-linearity such as local maxima or minima, and may include non-independent measures. Generalized additive mixed-modeling (GAMM provides an alternative description of dose-response that relaxes assumptions of independence and linearity. We compared GAMMs to traditional linear models for describing dose-response in IDMOC pharmacology studies.

  9. Differentiation of rat embryonic neural stem cells promoted by co-cultured Schwann cells

    Institute of Scientific and Technical Information of China (English)

    万虹; 安沂华; 张泽舜; 张亚卓; 王忠诚

    2003-01-01

    Objective To explore the factors which induce differentiation of embryonic neural stem cells. Methods Rat embryonic neural stem cells were co-cultured with newborn rat Schwann cells in serum-free medium. The phenotype and specific-markers including tubulin-β, glial fibrillary acidic protein (GFAP) and galactorcerebroside (GalC), were domonstrated by phase contrast microscopy and double immunofluorescence staining. Results Overall, 80%±5% of neural stem cells protruded several elongated processes and expressed tubulin-β antigen at high levels, while 20±3% of them protruded several short processes and were GalC or GFAP positive. Conclusion The factors secreted by Schwann cells could induce rat embryonic neural stem cell to differentiate.

  10. The regulation of bone turnover in ameloblastoma using an organotypic in vitro co-culture model

    Science.gov (United States)

    Eriksson, Tuula M; Day, Richard M; Fedele, Stefano; Salih, Vehid M

    2016-01-01

    Ameloblastoma is a rare, odontogenic neoplasm with benign histopathology, but extensive, local infiltrative capacity through the bone tissue it originates in. While the mechanisms of ameloblastoma invasion through the bone and bone absorption are largely unknown, recent investigations have indicated a role of the osteoprotegerin/receptor activator of nuclear factor kappa-B ligand regulatory mechanisms. Here, we present results obtained using a novel in vitro organotypic tumour model, which we have developed using tissue engineering techniques. Using this model, we analysed the expression of genes involved in bone turnover and detected a 700-fold increase in receptor activator of nuclear factor kappa-B ligand levels in the co-culture models with ameloblastoma cells cultured with bone cells. The model described here can be used for gene expression studies, as a basis for drug testing or for a more tailored platform for testing of the behaviour of different ameloblastoma tumours in vitro.

  11. Isolation of mammary epithelial cells from three-dimensional mixed-cell spheroid co-culture.

    Science.gov (United States)

    Xu, Kun; Buchsbaum, Rachel J

    2012-04-30

    -dimensional cultures of mixed cell populations (co-cultures)(16-22). With continued co-culture the cells form spheroids with the fibroblasts clustering in the interior and the epithelial cells largely on the exterior of the spheroids and forming multi-cellular projections into the matrix. Manipulation of the fibroblasts that leads to altered epithelial cell invasiveness can be readily quantified by changes in numbers and length of epithelial projections(23). Furthermore, we have devised a method for isolating epithelial cells out of three-dimensional co-culture that facilitates analysis of the effects of fibroblast exposure on epithelial behavior. We have found that the effects of co-culture persist for weeks after epithelial cell isolation, permitting ample time to perform multiple assays. This method is adaptable to cells of varying malignant potential and requires no specialized equipment. This technique allows for rapid evaluation of in vitro cell models under multiple conditions, and the corresponding results can be compared to in vivo animal tissue models as well as human tissue samples.

  12. Comparing three methods of co-culture of retinal pigment epithelium with progenitor cells derived human embryonic stem cells

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    Noushin Amirpour

    2013-01-01

    The adherent cells were morphologically examined using phase contrast microscopy and characterized by immunofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR Results: Evaluation of immunostaining showed that hESC, highly (>80% can be directed to the RPs fate. Upon co-culture of RPCs with RPE sheet using insert for 2 weeks or by the cell-to-cell contact, these cells differentiated to neural retina and expressed photoreceptor-specific markers. However, in direct co-culture, some mature photoreceptor markers like arrestin expressed in compare with indirect co-culture. Conclusions: The expression of late photoreceptor marker could be improved when RPE cells seeded on RPCs in compare with the use of insert.

  13. Liquid chromatography-mass spectrometry for metabolic footprinting of co-cultures of lactic and propionic acid bacteria

    DEFF Research Database (Denmark)

    Honore, Anders H.; Thorsen, Michael; Skov, Thomas

    2013-01-01

    Co-cultures of specific lactic and propionic acid bacteria have been shown to have an antagonistic effect against yeast and moulds in dairy systems. In studies of these co-cultures by bioassay-guided fractionation and analysis, numerous compounds have been reported to inhibit yeast and moulds...... could be a potent approach to elucidation of the mechanism. The purpose of this review is to discuss the two pre-requisites for such a study-the compound classes expected in the co-cultures, and on the basis of these, the most suitable analytical technique(s). Ultrahigh-performance liquid chromatography....... The strategy recommended in this study does not include all compound classes, and suggestions for supplementary methods are listed....

  14. Butanol production from alkali-pretreated rice straw by co-culture of Clostridium thermocellum and Clostridium saccharoperbutylacetonicum.

    Science.gov (United States)

    Kiyoshi, Keiji; Furukawa, Masataka; Seyama, Tomoko; Kadokura, Toshimori; Nakazato, Atsumi; Nakayama, Shunichi

    2015-06-01

    The co-culture of cellulolytic Clostridium thermocellum NBRC 103400 and butanol-producing Clostridium saccharoperbutylacetonicum strain N1-4 produced 5.5 g/L of butanol from 40 g/L of delignified rice straw pretreated with 1% (wt/vol) NaOH. The addition of cellulase (100 U/g biomass) in a co-culture system significantly increased butanol production to 6.9 g/L using 40 g/L of delignified rice straw. Compared to the control, this increase in butanol production was attributed to the enhancement of exoglucanase activity on lignocellulose degradation in experimental samples. The results showed that the co-culture system in conjunction with enhanced exoglucanase activity resulted in cost-effective butanol production from delignified rice straw. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Development of DARPP-32-positive parts of fetal pig ganglionic eminence and ventral mesencephalon in organotypic slice co-cultures

    DEFF Research Database (Denmark)

    Dall, Annette Møller; Rasmussen, Jens Zimmer

    2006-01-01

    Neurons from the fetal pig dopaminergic ventral mesencephalon (VM) and basal ganglia anlage (the ganglionic eminence) were co-cultured as organotypic slice cultures to study the development of the two interconnected brain areas. During a short developmental period (E35-E42), a groove separates th...... esterase (AChE) and were the preferred target areas for TH-positive fibers from the co-cultured VM....... (TH)-positive, dopaminergic fibers from co-cultured slices of the ventral mesencephalon. DARPP-32 expression was more extensive and dense in cultures of the lateral part of the striatal anlage than the medial part. The DARPP-32-positive areas moreover overlapped with areas rich in acetylcholine...

  16. The Effects of Neuregulin on Synapse Formation in Primary Muscle/Nerve Co-Cultures

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    Jessica Walsh

    2007-06-01

    Full Text Available In development, motor neurons innervate maturing myotubes to form the neuromuscular junction (NMJ. During this process, the nerve terminal secretes a protein called neuregulin (NRG1. NRG1 acts as a signal which increases the number of acetylcholine receptors (AChRs on the postsynaptic membrane. NRG1 binds to receptors on the surface of the muscle, known as erbB receptors. The binding of NRG1 causes the erbB receptor to auto-phosphorylate (Fu, 1999. As a result, there is an increase in transcription of the gene for AchRs, integral membrane proteins that respond to the binding of the neurotransmitter acetylcholine. In order to study the effects of NRG1 on early stages of synapse formation, we prepared co-cultures of dissociated muscle cells from postnatal day 1 (P1 mice and neural tube explants from embryonic day 11 (E11 mice. Silicone chambers were created as a system for growing dissociated muscle cells and neuronal explants in co-culture (Loeb, 1999. ErbB inhibitor (PD 158780 was added to chambers prior to the formation of the NMJ. After one week the tissue was fixed and stained to visualize the synapses. Based on the results of two experiments, the chambers that were not treated with the inhibitor had an average of 10 times more AchR (+ contacts. Therefore, at the time point studied, it appears that NRG1 signaling through the erbB receptor tyrosine kinases is necessary for the formation of AchR (+ receptor contacts at the motor terminal in the post-synaptic membrane.

  17. Porphyromonas gingivalis displays a competitive advantage over Aggregatibacter actinomycetemcomitans in co-cultured biofilm.

    Science.gov (United States)

    Takasaki, K; Fujise, O; Miura, M; Hamachi, T; Maeda, K

    2013-06-01

    Biofilm formation occurs through the events of cooperative growth and competitive survival among multiple species. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans are important periodontal pathogens. The aim of this study was to demonstrate competitive or cooperative interactions between these two species in co-cultured biofilm. P. gingivalis strains and gingipain mutants were cultured with or without A. actinomycetemcomitans. Biofilms formed on glass surfaces were analyzed by crystal violet staining and colony counting. Preformed A. actinomycetemcomitans biofilms were treated with P. gingivalis culture supernatants. Growth and proteolytic activities of gingipains were also determined. Monocultured P. gingivalis strains exhibited a range of biofilm-formation abilities and proteolytic activities. The ATCC33277 strain, noted for its high biofilm-formation ability and proteolytic activity, was found to be dominant in biofilm co-cultured with A. actinomycetemcomitans. In a time-resolved assay, A. actinomycetemcomitans was primarily the dominant colonizer on a glass surface and subsequently detached in the presence of increasing numbers of ATCC33277. Detachment of preformed A. actinomycetemcomitans biofilm was observed by incubation with culture supernatants from highly proteolytic strains. These results suggest that P. gingivalis possesses a competitive advantage over A. actinomycetemcomitans. As the required biofilm-formation abilities and proteolytic activities vary among P. gingivalis strains, the diversity of the competitive advantage is likely to affect disease recurrence during periodontal maintenance. © 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Using a co-culture microsystem for cell migration under fluid shear stress.

    Science.gov (United States)

    Yeh, Chia-Hsien; Tsai, Shen-Hsing; Wu, Li-Wha; Lin, Yu-Cheng

    2011-08-01

    We have successfully developed a microsystem to co-cultivate two types of cells with a minimum defined gap of 50 μm, and to quantitatively study the impact of fluid shear stress on the mutual influence of cell migration velocity and distance. We used the hydrostatic pressure to seed two different cells, endothelial cells (ECs) and smooth muscle cells (SMCs), on opposite sides of various gap sizes (500 μm, 200 μm, 100 μm, and 50 μm). After cultivating the cells for 12 h and peeling the co-culture microchip from the culture dish, we studied the impacts of gap size on the migration of either cell type in the absence or presence of fluid shear stress (7 dyne cm(-2) and 12 dyne cm(-2)) influence. We found that both gap size and shear stress have profound influence on cell migration. Smaller gap sizes (100 μm and 50 μm) significantly enhanced cell migration, suggesting a requirement of an effective concentration of released factor(s) by either cell type in the gap region. Flow-induced shear stress delayed the migration onset of either cell type in a dose-dependent manner regardless of the gap size. Moreover, shear stress-induced decrease of cell migration becomes evident when the gap size was 500 μm. We have developed a co-culture microsystem for two kinds of cells and overcome the conventional difficulties in observation and mixed culture, and it would have more application for bio-manipulation and tissue repair engineering.

  19. Electrolytic valving isolation of cell co-culture microenvironment with controlled cell pairing ratios.

    Science.gov (United States)

    Chen, Yu-Chih; Ingram, Patrick; Yoon, Euisik

    2014-12-21

    Cancer-stromal interaction is a critical process in tumorigenesis. Conventional dish-based co-culture assays simply mix two cell types in the same dish; thus, they are deficient in controlling cell locations and precisely tracking single cell behavior from heterogeneous cell populations. Microfluidic technology can provide a good spatial-temporal control of microenvironments, but the control has been typically realized by using external pumps, making long-term cultures cumbersome and bulky. In this work, we have presented a cell-cell interaction microfluidic platform that can accurately control the co-culture microenvironment by using a novel electrolytic cell isolation scheme without using any valves or pneumatic pumps. The proposed microfluidic platform can also precisely control the number of interacting cells and pairing ratios to emulate cancer niches. More than 80% of the chambers captured the desired number of cells. The duration of cell isolation can be adjusted by electrolytic bubble generation and removal. We have verified that the electrolytic process has a negligible effect on cell viability and proliferation in our platform. To the best of our knowledge, this work is the first attempt to incorporate electrolytic bubble generation as a cell isolation method in microfluidics. For proof of feasibility, we have performed cell-cell interaction assays between prostate cancer (PC3) cells and myoblast (C2C12) cells. The preliminary results demonstrated the potential of using electrolysis for micro-environmental control during cell culture. Also, the ratio controlled cell-cell interaction assays were successfully performed which showed that the cell pairing ratios of PC3 to C2C12 affected the proliferation rate of myoblast cells due to increased secretion of growth factors from prostate cancer cells.

  20. Inference of Interactions in Cyanobacterial-Heterotrophic Co-Cultures via Transcriptome Sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Beliaev, Alex S.; Romine, Margaret F.; Serres, Margaret; Bernstein, Hans C.; Linggi, Bryan E.; Markillie, Lye Meng; Isern, Nancy G.; Chrisler, William B.; Kucek, Leo A.; Hill, Eric A.; Pinchuk, Grigoriy; Bryant, Donald A.; Wiley, H. S.; Fredrickson, Jim K.; Konopka, Allan

    2014-04-29

    We employed deep sequencing technology to identify transcriptional adaptation of the euryhaline unicellular cyanobacterium Synechococcus sp. PCC 7002 and the marine facultative aerobe Shewanella putrefaciens W3-18-1 to growth in a co-culture and infer the effect of carbon flux distributions on photoautotroph-heterotroph interactions. The overall transcriptome response of both organisms to co-cultivation was shaped by their respective physiologies and growth constraints. Carbon limitation resulted in the expansion of metabolic capacities which was manifested through the transcriptional upregulation of transport and catabolic pathways. While growth coupling occurred via lactate oxidation or secretion of photosynthetically fixed carbon, there was evidence of specific metabolic interactions between the two organisms. On one hand, the production and excretion of specific amino acids (methionine and alanine) by the cyanobacterium correlated with the putative downregulation of the corresponding biosynthetic machinery of Shewanella W3-18-1. On the other hand, the broad and consistent decrease of mRNA levels for many Fe-regulated Synechococcus 7002 genes during co-cultivation suggested increased Fe availability as well as more facile and energy-efficient mechanisms for Fe acquisition by the cyanobacterium. Furthermore, evidence pointed at potentially novel interactions between oxygenic photoautotrophs and heterotrophs related to the oxidative stress response as transcriptional patterns suggested that Synechococcus 7002 rather than Shewanella W3-18-1 provided scavenging functions for reactive oxygen species under co-culture conditions. This study provides an initial insight into the complexity of photoautotrophic-heterotrophic interactions and brings new perspectives of their role in the robustness and stability of the association.

  1. Expression profile of germ stem cell-specific genes in human spermatogonial stem cells after co culture with sertoli cells

    Directory of Open Access Journals (Sweden)

    Maria Zahiri

    2014-05-01

    Full Text Available Background: Human spermatogonial stem cells (SSCs, are the foundation of spermatogenesis. Because of low number and lack of significant marker in human SSCs, studying their characteristics, could provide better understanding about the biology of male fertility. This study was designed to examine the effects of in vitro co-culture with sertoli cells on SSC colonization and germ cells specific gene expression of human spermatogonial stem cells. Material and Methods: Testicular cells were isolated from testis biopsies by using two step enzymatic digestion and differential plating. two culture system were designed: co-culture with patient Sertoli cells and culture of SSC without co-culture(as control group. The number and diameter of colonies were evaluated during 3 weeks of culture. The expression of alpha 6 integrin, beta1 integrin and PLZF, as germ stem cell specific markers, was assessed using quantitative RT-PCR. Statistical analysis was performed using one way ANOVA in SPSS vesion 16 software with 95% Confidence interval . Result: Our results were showed that the number and diameter of colonies increased significantly in co-culture with sertoli cells (P<0.05. The expression profile of genes in 2nd and 3rd weeks of culture revealed that there is significant higher expression of germ stem cell markers in our co-culture group versus control group. Conclusion: Based on the optimal effects of sertoli cells on spermatogonial stem cells, co culture of the human SSCs with the feeder layer sertoli may be used as a suitable method for the enrichment of human spermatogonial stem cells.

  2. Development of an Insert Co-culture System of Two Cellular Types in the Absence of Cell-Cell Contact.

    Science.gov (United States)

    Renaud, Justine; Martinoli, Maria-Grazia

    2016-07-17

    The role of secreted soluble factors in the modification of cellular responses is a recurrent theme in the study of all tissues and systems. In an attempt to make straightforward the very complex relationships between the several cellular subtypes that compose multicellular organisms, in vitro techniques have been developed to help researchers acquire a detailed understanding of single cell populations. One of these techniques uses inserts with a permeable membrane allowing secreted soluble factors to diffuse. Thus, a population of cells grown in inserts can be co-cultured in a well or dish containing a different cell type for evaluating cellular changes following paracrine signaling in the absence of cell-cell contact. Such insert co-culture systems offer various advantages over other co-culture techniques, namely bidirectional signaling, conserved cell polarity and population-specific detection of cellular changes. In addition to being utilized in the field of inflammation, cancer, angiogenesis and differentiation, these co-culture systems are of prime importance in the study of the intricate relationships that exist between the different cellular subtypes present in the central nervous system, particularly in the context of neuroinflammation. This article offers general methodological guidelines in order to set up an experiment in order to evaluating cellular changes mediated by secreted soluble factors using an insert co-culture system. Moreover, a specific protocol to measure the neuroinflammatory effects of cytokines secreted by lipopolysaccharide-activated N9 microglia on neuronal PC12 cells will be detailed, offering a concrete understanding of insert co-culture methodology.

  3. Contributing factors in the improvement of cellulosic H2 production in Clostridium thermocellum/Thermoanaerobacterium co-cultures.

    Science.gov (United States)

    Wang, Mingyu; Zhao, Qi; Li, Ling; Niu, Kangle; Li, Yi; Wang, Fangzhong; Jiang, Baojie; Liu, Kuimei; Jiang, Yi; Fang, Xu

    2016-10-01

    Lignocellulosic biohydrogen is a promising renewable energy source that could be a potential alternative to the unsustainable fossil fuel-based energy. Biohydrogen production could be performed by Clostridium thermocellum that is the fastest known cellulose-degrading bacterium. Previous investigations have shown that the co-culture of C. thermocellum JN4 and a non-cellulolytic bacterium Thermoanaerobacterium thermosaccharolyticum GD17 produces more hydrogen than the C. thermocellum JN4 mono-culture, but the mechanism of this improvement is unknown. In this work, we carried out genomic and evolutionary analysis of hydrogenase-coding genes in C. thermocellum and T. thermosaccharolyticum, identifying one Ech-type [NiFe] hydrogenase complex in each species, and, respectively, five and four monomeric or multimeric [FeFe] hydrogenases in the two species. Further transcriptional analysis showed hydrogenase-coding genes in C. thermocellum are regulated by carbon sources, while hydrogenase-coding genes in T. thermosaccharolyticum are not. However, comparison between transcriptional abundance of hydrogenase-coding genes in mono- and co-cultures showed the co-culturing condition leads to transcriptional changes of hydrogenase-coding genes in T. thermosaccharolyticum but not C. thermocellum. Further metabolic analysis showed T. thermosaccharolyticum produces H2 at a rate 4-12-fold higher than C. thermocellum. These findings lead to the suggestion that the improvement of H2 production in the co-culture over mono-culture should be attributed to changes in T. thermosaccharolyticum but not C. thermocellum. Further suggestions can be made that C. thermocellum and T. thermosaccharolyticum perform highly specialized tasks in the co-culture, and optimization of the co-culture for more lignocellulosic biohydrogen production should be focused on the improvement of the non-cellulolytic bacterium.

  4. Effects of pyridoxine on rat testes by means of Sertoli-germ cell co-culture system in vitro

    Institute of Scientific and Technical Information of China (English)

    Huang Houjin

    2001-01-01

    Objective To investigate the effects of pyridoxine on rat testis in vitro. Method an in vitro systen of Sertoligem cell co-culture was applied, the toxic effects of pyridoxine at different concentrations an exposed duration were olserved. Results The detachment of germ cells from sertoli cells showed marked dose-response and time response relafionships with the exposure of pyridoxine. Meanwhile, the characteristic of loosing and ratracting skeletun in the Sertoli cells was found. Conclusions The effects induced by pyridoxine in vitro may reflect damage to Sertoli cells, and testicular cells co-culture could be of value for the study of underlying mechanisms of toxic effects of pyridoxine on rat testis.

  5. Genome sequencing and annotation of Aeromonas sp. HZM

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    Patric Chua

    2015-09-01

    Full Text Available We report the draft genome sequence of Aeromonas sp. strain HZM, isolated from tropical peat swamp forest soil. The draft genome size is 4,451,364 bp with a G + C content of 61.7% and contains 10 rRNA sequences (eight copies of 5S rRNA genes, single copy of 16S and 23S rRNA each. The genome sequence can be accessed at DDBJ/EMBL/GenBank under the accession no. JEMQ00000000.

  6. Effect of solar irradiation on extracellular enzymes of Aeromonas proteolytica

    Science.gov (United States)

    Foster, B. G.

    1973-01-01

    The bacterium Aeromonas proteolytica was selected for studying the effects of solar irradiation on extracellular enzymes because it produces an endopeptidase that is capable of degrading proteins and a hemolysin that is active in lysing human erythrocytes. Possible alterations in the rate of enzyme production in response to the test conditions are currently underway and are not available for this preliminary report. Completed viability studies are indicative that little difference exists among the survival curves derived for cells exposed to various components of ultraviolet irradiation in space.

  7. Multifocal Aeromonas Osteomyelitis in a Child with Leukemia

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    Dimitrios Doganis

    2016-01-01

    Full Text Available Aeromonas hydrophila is a Gram negative organism causing both intestinal and extraintestinal disease. The case of a 14-year-old girl with underlying immunodeficiency and leukemia who developed systemic A. hydrophila infection is described in this report. While in deep bone marrow aplasia she developed fever, severe pain in the lower extremities, and swelling of the left femur. Blood culture showed Escherichia coli and A. hydrophila whereas pus culture from the soft tissue swelling showed the presence of A. hydrophila. Imaging studies showed diffuse osteolytic lesions. Patient received 5 months of intravenous and oral antibiotics and she improved clinically whereas the radiology findings persisted.

  8. Use of Aeromonas as a process indicator during swine carcass dressing and cutting

    Science.gov (United States)

    Palumbo, Samuel A.; Yu, Linda S. L.

    1999-01-01

    Using starch ampicillin agar, qualitative and quantitative determinations of Aeromonas spp. were made at several sites during swine carcass dressing and cutting. Aeromonas spp. were observed at all sites surveyed. Levels increased during shackling and passage through the first and middle polisher/washers, and significantly decreased during the singeing steps. Passage through the final polisher/washer caused a small increase in levels in Aeromonas spp. and these levels then remained constant during the rest of the carcass dressing operation. Aeromonas spp. were also isolated from the room where the carcasses were cut into wholesale cuts and cuts for further processing. Presumptive Aeromonas spp. cultures isolated from the different sites were confirmed as belonging to the genus Aeromonas and then speciated using the biochemical scheme of Joseph and Carnahan; 81% of the cultures were identified at A. hydrophila. Since most isolates were A. hydrophila, determination of the origin of isolates from different sites in the processing plant must await utilizing molecular biotyping techniques on the cultures. These results indicate the Aeromonas spp. occurs extensively in the swine dressing environment and thus represents a possible public health hazard and potential spoilage concern. Changes in cleaning and sanitizing of equipment may be necessary during swine carcass dressing and cutting to guard against this pathogen.

  9. Detection of antibiotic resistance, virulence gene determinants and biofilm formation in Aeromonas species isolated from cattle.

    Science.gov (United States)

    Igbinosa, Isoken H; Igbinosa, Etinosa O; Okoh, Anthony I

    2015-11-01

    This study aimed to assess the antibiogram of Aeromonas strains recovered from cattle faeces and the potential pathogenic status of the isolates. The antibiogram of the Aeromonas isolates demonstrated total resistance to clindamycin oxacillin, trimethoprim, novobiocin and ticarcillin. However, Aeromonas strains were sensitive to cefotaxime, oxytetracycline and tobramycin. The Aeromonas strains from Lovedale and Fort Cox farms were found to possess some virulence genes. The percentage distribution was aer 71.4%, ast 35.7%, fla 60.7%, lip 35.7% and hlyA 25% for Lovedale farm and aer 63.1%, alt 10.5%, ast 55.2%, fla 78.9%, lip 21% and hlyA 35.9% for Fort Cox farm. Class 1 integron was present in 27% of Aeromonas isolates; the bla TEM gene was present in 34.8%, while the blaP1 class A β-lactamase gene was detected in 12.1% of the isolates. Approximately 86% of the isolates formed a biofilm on microtitre plates. The presence of multiple antibiotic resistance and virulence genes in Aeromonas isolates from cattle faeces reveals the pathogenic and infectious importance of these isolates and is of great significance to public health. The possession of a biofilm-forming capability by such isolates may lead to difficulty during the management of infection related to Aeromonas species.

  10. Estereologia das glândulas bulbouretrais do coelho (Oryctolagus cuniculus e da cobaia (Cavia porcellus

    Directory of Open Access Journals (Sweden)

    Bélgica Vásquez

    2014-12-01

    Full Text Available As glândulas bulbouretrais (GBU no coelho (Oryctolagus cuniculus e na cobaia (Cavia porcellus desempenham um papel importante na fisiologia reprodutiva. No entanto, seus aspectos histológico e estereológico são escassos. Assim, o objetivo desta pesquisa foi comparar características estereológicas entre as GBU do coelho e da cobaia como um primeiro passo para a compreensão das variáveis morfométricas que participam nos processos reprodutivos. Foram utilizados 5 coelhos e 5 cobaias adultos machos, saudáveis, obtidos do Biotério da Universidade de La Frontera, Temuco, Chile. A região pélvica foi dissecada isolando-se a GBU de cada animal. Determinou-se o peso e o volume de cada glândula. Estas foram fixadas em formalina tamponada durante 24 horas e incluídas em paraplast. Cortes seriados de 4 μm de espessura foram corados com HE para análise estereológica. A média de células glandulares na GBU do coelho foi 19,50 x 10(5mm³ (DP 2,35 e da cobaia 10,57 x 10(5mm³ (DP 2,07 e a porcentagem média de tecido glandular foi de 25,52% (DP 2,20 e 17,20% (DP 3,33, respectivamente. Todos os parâmetros estereológicos comparados tiveram uma diferença estatisticamente significativa (p<0,0001. Estas diferenças poderiam ser explicadas porque há maior proximidade celular do epitélio secretor, menor diâmetro do lúmen dos ácinos e da relação núcleo citoplasma na GBU do coelho. Assim, os ácinos da GBU apresentam maior quantidade de células por mm³ do que na GBU do coelho. Estes parâmetros podem ser influenciados por fatores hormonais, etários, sazonais e ambientais, entre outros. Considerar as características morfológicas da GBU nesses animais poderia condicionar o êxito da reprodução por parte do macho.

  11. Permeability of PEGylated Immunoarsonoliposomes Through In Vitro Blood Brain Barrier-Medulloblastoma Co-culture Models for Brain Tumor Therapy

    NARCIS (Netherlands)

    Al-Shehri, A.; Favretto, M.E.; Ioannou, P.V.; Romero, I.A.; Couraud, P.O.; Weksler, B.B.; Parker, T.L.; Kallinteri, P.

    2015-01-01

    PURPOSE: Owing to restricted access of pharmacological agents into the brain due to blood brain barrier (BBB) there is a need: 1. to develop a more representative 3-D-co-culture model of tumor-BBB interaction to investigate drug and nanoparticle transport into the brain for diagnostic and therapeuti

  12. Interplay between human microglia and neural stem/progenitor cells in an allogeneic co-culture model.

    Science.gov (United States)

    Liu, Jia; Hjorth, Erik; Zhu, Mingqin; Calzarossa, Cinzia; Samuelsson, Eva-Britt; Schultzberg, Marianne; Åkesson, Elisabet

    2013-11-01

    Experimental neural cell therapies, including donor neural stem/progenitor cells (NPCs) have been reported to offer beneficial effects on the recovery after an injury and to counteract inflammatory and degenerative processes in the central nervous system (CNS). The interplay between donor neural cells and the host CNS still to a large degree remains unclear, in particular in human allogeneic conditions. Here, we focused our studies on the interaction of human NPCs and microglia utilizing a co-culture model. In co-cultures, both NPCs and microglia showed increased survival and proliferation compared with mono-cultures. In the presence of microglia, a larger subpopulation of NPCs expressed the progenitor cell marker nestin, whereas a smaller group of NPCs expressed the neural markers polysialylated neural cell adhesion molecule, A2B5 and glial fibrillary acidic protein compared with NPC mono-cultures. Microglia thus hindered differentiation of NPCs. The presence of human NPCs increased microglial phagocytosis of latex beads. Furthermore, we observed that the expression of CD200 molecules on NPCs and the CD200 receptor protein on microglia was enhanced in co-cultures, whereas the release of transforming growth factor-β was increased suggesting anti-inflammatory features of the co-cultures. To conclude, the interplay between human allogeneic NPCs and microglia, significantly affected their respective proliferation and phenotype. Neural cell therapy including human donor NPCs may in addition to offering cell replacement, modulate host microglial phenotypes and functions to benefit neuroprotection and repair.

  13. Improving vascularization of engineered bone through the generation of pro-angiogenic effects in co-culture systems.

    Science.gov (United States)

    Unger, Ronald E; Dohle, Eva; Kirkpatrick, C James

    2015-11-01

    One of the major problems with bone tissue engineering is the development of a rapid vascularization after implantation to supply the growing osteoblast cells with the nutrients to grow and survive as well as to remove waste products. It has been demonstrated that capillary-like structures produced in vitro will anastomose rapidly after implantation and become functioning blood vessels. For this reason, in recent years many studies have examined a variety of human osteoblast and endothelial cell co-culture systems in order to distribute osteoblasts on all parts of the bone scaffold and at the same time provide conditions for the endothelial cells to migrate to form a network of capillary-like structures throughout the osteoblast-colonized scaffold. The movement and proliferation of endothelial cells to form capillary-like structures is known as angiogenesis and is dependent on a variety of pro-angiogenic factors. This review summarizes human 2- and 3-D co-culture models to date, the types and origins of cells used in the co-cultures and the proangiogenic factors that have been identified in the co-culture models.

  14. Co-Culture of Early Embryo with Human Decidual Stromal Cells in vitro by Improvement of Early Embryo Development

    Institute of Scientific and Technical Information of China (English)

    YAN Jie; ZHU Guijin; LIU Jianxin; AI Jihui

    2000-01-01

    An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM+0.4%bovine serum albumin (BSA) and 163 embryos cultured in MEM+15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73% developed to the morula stage and 67.21% cavitated to blastocysts with 59.74 % hatching, as compared with 61.34% to morula stage, 48.47% to blastocysts and none hatching in the controls,respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.

  15. Multi-strain co-cultures surpass blends for broad spectrum biological control of maladies of potatoes in storage

    Science.gov (United States)

    Pseudomonas fluorescens strains S11:P:12, P22:Y:05, and S22:T:04 have been documented to suppress four important problems in potato storages — dry rot, late blight, pink rot, and sprouting. This research investigates the efficacy and consistency of strain mixtures produced by co-culturing strains t...

  16. Impact of Co-Culturing with Fractionated Carbon-Ion-Irradiated Cancer Cells on Bystander Normal Cells and Their Progeny.

    Science.gov (United States)

    Autsavapromporn, Narongchai; Liu, Cuihua; Konishi, Teruaki

    2017-09-01

    The purpose of this study was to compare the biological effects of fractionated doses versus a single dose of high-LET carbon ions in bystander normal cells, and determine the effect on their progeny using the layered tissue co-culture system. Briefly, confluent human glioblastoma (T98G) cells received a single dose of 6 Gy or three daily doses of 2 Gy carbon ions, which were then seeded on top of an insert with bystander normal skin fibroblasts (NB1RGB) growing underneath. Cells were co-cultured for 6 h or allowed to grow for 20 population doublings, then harvested and assayed for different end points. A single dose of carbon ions resulted in less damage in bystander normal NB1RGB cells than the fractionated doses. In contrast, the progeny of bystander NB1RGB cells co-cultured with T98G cells exposed to fractionated doses showed less damage than progeny from bystander cells co-cultured with single dose glioblastoma cells. Furthermore, inhibition of gap junction communication demonstrated its involvement in the stressful effects in bystander cells and their progeny. These results indicate that dose fractionation reduced the late effect of carbon-ion exposure in the progeny of bystander cells compared to the effect in the initial bystander cells.

  17. Co-culturing Chlorella minutissima with Escherichia coli can increase neutral lipid production and improve biodiesel quality.

    Science.gov (United States)

    Higgins, Brendan T; Labavitch, John M; VanderGheynst, Jean S

    2015-09-01

    Lipid productivity and fatty acid composition are important metrics for the production of high quality biodiesel from algae. Our previous results showed that co-culturing the green alga Chlorella minutissima with Escherichia coli under high-substrate mixotrophic conditions enhanced both culture growth and crude lipid content. To investigate further, we analyzed neutral lipid content and fatty acid content and composition of axenic cultures and co-cultures produced under autotrophic and mixotrophic conditions. We found that co-culturing C. minutissima with E. coli under high substrate conditions (10 g/L) increased neutral lipid content 1.9- to 3.1-fold and fatty acid content 1.5- to 2.6-fold compared to equivalent axenic C. minutissima cultures. These same co-cultures also exhibited a significant fatty acid shift away from trienoic and toward monoenoic fatty acids thereby improving the quality of the synthesized fatty acids for biodiesel production. Further investigation suggested that E. coli facilitates substrate uptake by the algae and that the resulting growth enhancement induces a nitrogen-limited condition. Enhanced carbon uptake coupled with nitrogen limitation is the likely cause of the observed neutral lipid accumulation and fatty acid profile changes.

  18. [AEROMONAS BACTERIA ISOLATED FROM BITHYNIIDAE MOLLUSKS AND THEIR HABITATS: SPECIES COMPOSITION AND BIOLOGICAL PROPERTIES. COMMUNICATION 1].

    Science.gov (United States)

    Stepanova, T F; Bukharin, O V; Kataeva, L V; Perunova, N B; Karpukhina, N F

    2015-01-01

    The purpose of this investigation was to study the species composition and biological properties of Aeromonas bacteria isolated from Bithyniidae mollusks and their habitat (a water reservoir). The Bithyniidae mollusks and water from their habitat were the material to be studied. A total of 176 Aeromonas strains were isolated from the mollusks and water. A. veronii, A. hydrophila, and A. ichthiosmia were most common in the mollusks and A. veronii and A. ichthiosmia were in the water. All the strains isolated had hemolytic activity and no lysozyme or plasma coagulase activity. The magnitude of lecithinase and antilysozymic activities and biofilm formation of the Aeromonas bacteria varied with the isolation source of their strains.

  19. Aeromonas salmonicida bacteremia associated with chronic well water consumption in a patient with diabetes

    Directory of Open Access Journals (Sweden)

    Christine Ann Moore

    2017-01-01

    Full Text Available Aeromonas salmonicida is associated with superficial skin infections in fish. Its virulence factors allow colonization of water including surface water such as salt water, beaches, and fresh water wells. Moreover, it is possible for immunocompromised patients to develop invasive disease after chronic exposure to Aeromonas spp. through contaminated water. While there are reports of Aeromonas spp. bacteremia following water ingestion, there have been no reports of A. salmonicida bacteremia from water consumption. We report the first case of A. salmonicida bacteremia in a patient with diabetes due to chronic consumption of well water.

  20. Lippia alba essential oil promotes survival of silver catfish (Rhamdia quelen) infected with Aeromonas sp.

    OpenAIRE

    FERNANDO J. SUTILI; Cunha,Mauro A.; Rosangela E. Ziech; Krewer,Carina C.; CARLA C. ZEPPENFELD; Heldwein, Clarissa G.; Gressler, Leticia T.; Berta M. Heinzmann; Agueda C. de Vargas; BERNARDO BALDISSEROTTO

    2015-01-01

    In vitro and in vivo activity of the Lippia alba essential oil (EO) against Aeromonas sp. was evaluated. In the in vitro assay the minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of EO for Aeromonas cells were determined using the microdilution method. Twenty five strains of Aeromonas sp. isolated from infected fish obtained from local fish farms were used. MIC and MBC values were 2862 and 5998 µg mL-1 for L. alba EO and 0.5 and 1.2 µg mL-1 for ...

  1. Necrotizing Fasciitis Secondary to Aeromonas Infection Presenting with Septic Shock

    Directory of Open Access Journals (Sweden)

    Nikhil Bhatia

    2017-01-01

    Full Text Available This report describes a case of necrotizing fasciitis presenting with septic shock due to an Aeromonas infection. The patient cut his foot while mowing the lawn and then spent time in a pool with black mold. He began feeling ill and developed swelling and a quarter-sized black area on his right lower extremity. Despite being hemodynamically unstable with systolic blood pressure in the low 70s, the patient was transferred to our facility from outside hospital 100 miles away. Upon arriving to facility, the patient appeared to be septic and the infected area of skin had grown. Irrigation and debridement were performed and appropriate antibiotic therapy was given; however, the patient subsequently died on hospital day 8. On review of the literature, cases of necrotizing fasciitis due to Aeromonas infection have been treated successfully with the aforementioned therapy; however, there is high mortality associated with these infections, many times related to a delayed diagnosis. Our patient also had multiple poor prognostic factors including hepatic dysfunction and immunosuppression.

  2. Incidence and toxigenicity of Aeromonas hydrophila in seafood.

    Science.gov (United States)

    Tsai, G J; Chen, T H

    1996-08-01

    Three selective media, Oxoid Aeromonas agar (OA), blood ampicillin agar (BA) and starch ampicillin agar (SA) were used to evaluate the presence of Aeromonas hydrophila in 66 samples of oyster, shrimp, fish and surimi products. Oyster had the highest incidence, with 50% positive, whilst no A. hydrophila was found in the surimi. Of the three selective media, BA displayed the highest recovery rate of A. hydrophila from seafood. Forty-eight isolates from this survey were tested for their capability to produce hemolysin and cytotoxin. Hemolysin was produced by 79.2% of the isolates and cytotoxin was produced by 91.7% of the isolates in brain heart infusion broth. One of the toxin-producing isolates from oyster, strain 8-169, was further tested for growth and toxin production in oyster, shrimp and fish at various temperatures. This particular isolate grew best and had highest toxin production in oyster. Hemolysin and cytotoxin were produced earlier at 28 degrees C than at 37 degrees C, and titers of hemolysin were also higher at 28 degrees C. At 5 degrees C, it was able to grow and produce hemolysin in oyster.

  3. Colonization of streptomycin-treated mice by Aeromonas species.

    Science.gov (United States)

    Sanderson, K; Ghazali, F M; Kirov, S M

    1996-03-01

    Streptomycin-treated adult mice were investigated as a possible model for studying the enteropathogenicity of Aeromonas species. C57BL mice pre-treated with streptomycin (5.0 g/L drinking water, 48 hours) received a single intragastric dose (10(10) bacteria /10.5 mL) of one of six well-characterized, toxin-producing, human diarrhoeal isolates of A. veronii biovar sobria (n = 3) or A. hydrophila (n = 3). Their faeces were examined for Aeromonas for 10 days post-challenge. All strains colonized the antibiotic-treated mice. Colonization did not occur in mice which did not receive streptomycin. Strains of A. hydrophila were recovered in greater numbers than strains of A. veronii biovar sobria, and colonized ( > or = 10(3) cfu/g of faeces) a greater proportion of mice at day 10. Strains of the latter species, however, were more adherent in cell line assays used as models of intestinal adhesion. A. hydrophila strains localized in the large intestine and appeared not to be cell associated. This study, therefore, points to species-related differences in intestinal colonization mechanisms. The streptomycin-treated adult mouse model may prove useful for further investigation of some of these mechanisms. Diarrhoeal symptoms were, however, not produced in this model.

  4. Effects of the probiotic Enterococcus faecium NCIMB 10415 on selected lactic acid bacteria and enterobacteria in co-culture.

    Science.gov (United States)

    Starke, I C; Zentek, J; Vahjen, W

    2015-01-01

    Enterococcus faecium NCIMB 10415 is used as a probiotic for piglets and has been shown to modify the porcine intestinal microbiota. However, the mode of action of this probiotic modification is still unclear. One possible explanation is the direct growth inhibiting or stimulating effect of the probiotic on other indigenous bacteria. Therefore, the aim of the present study was to examine the growth interactions of the probiotic with different indigenous porcine bacteria in vitro. Reference strains were cultivated with the probiotic E. faecium strain NCIMB10415 (SF68) in a checkerboard assay with 102 to 105 cells/ml inoculum per strain. Growth kinetics were recorded for 8 h and used to determine specific growth of the co-cultures. Additionally, total DNA was extracted from the co-cultures at the end of the incubation to verify which strain in the co-culture was affected. Co-cultivation with eight Enterococcus spp. tester strains showed strain-specific growth differences. Three of four E. faecium strains were not influenced by the probiotic strain. PCR results showed reduced growth of the probiotic strain in co-culture with E. faecium DSM 6177. Three of four Enterococcus faecalis strains showed reduced specific growth in co-culture with the probiotic strain. However, E. faecalis DSM 20478 impaired growth of the probiotic E. faecium strain. The growth of Lactobacillus johnsonii DSM 10533 and Lactobacillus reuteri DSM 20016 was enhanced in co-culture with the probiotic strain, but co-cultivations with Lactobacillus mucosae DSM13345 or Lactobacillus amylovorus DSM10533 showed no differences. Co-cultures with the probiotic E. faecium showed no impact on the growth rate of four different enterobacterial reference strains (2 strains of Salmonella enterica and 2 strains of Escherichia coli), but PCR results showed reduced cell numbers for a pathogenic E. coli isolate at higher concentration of the probiotic strain. As the in vitro effect of the probiotic E. faecium on

  5. Defining conditions for the co-culture of Caco-2 and HT29-MTX cells using Taguchi design.

    Science.gov (United States)

    Chen, Xiu-Min; Elisia, Ingrid; Kitts, David D

    2010-01-01

    The co-culture of Caco-2 and HT29 cells for testing intestinal drug and nutrient transport and metabolism provides the presence of both absorptive and goblet cells, both of which have different culture requirements for optimal growth and function. The research on the co-culture of Caco-2 and HT29 cells is very limited in respect to refining specific conditions that reduce intra- and inter-laboratory variations. In the present study we reported conditions that enable reproducible results to be obtained for drug permeability using in vitro co-culture of Caco-2 and HT29-MTX based on Taguchi experimental design. The selection of four factors that specified cell culture conditions, namely culture medium, seeding time, seeding density, and Caco-2:HT29-MTX ratio on TEER value and individual permeability coefficients of propranolol, ketoprofen and furosemide was established. Based on the selected conditions for co-culture, we also confirmed the functionality of the final chosen culture condition using nitric oxide as an indicator of intestinal inflammation. Choice of cell culture time and culture medium represented two of the most important factors that affected TEER values and the permeability coefficients of the model drugs. On the other hand, the seeding density and the Caco-2:HT29-MTX ratio exerted no significant influence on TEER values and the drug permeability coefficients. No absolute optimal cell culture condition could be obtained for all drugs; however subsequent confirmation experiments concluded that excellent precision for TEER values and drug permeability coefficients was obtained from the two operators using the following combination of conditions, namely an initial seeding density of 1 x 10(5) Caco-2 and HT29-MTX cells/cm(2) at a ratio of 9:1, followed by a 21day culture time in MEM medium. Finally, functionality of the co-culture model system using the above selected in vitro conditions resulted in comparable nitric oxide synthesis to that of a Caco-2

  6. A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity

    Science.gov (United States)

    Anderl, Janet L; Redpath, Stella; Ball, Andrew J

    2009-01-01

    High Content Analysis (HCA) assays combine cells and detection reagents with automated imaging and powerful image analysis algorithms, allowing measurement of multiple cellular phenotypes within a single assay. In this study, we utilized HCA to develop a novel assay for neurotoxicity. Neurotoxicity assessment represents an important part of drug safety evaluation, as well as being a significant focus of environmental protection efforts. Additionally, neurotoxicity is also a well-accepted in vitro marker of the development of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Recently, the application of HCA to neuronal screening has been reported. By labeling neuronal cells with βIII-tubulin, HCA assays can provide high-throughput, non-subjective, quantitative measurements of parameters such as neuronal number, neurite count and neurite length, all of which can indicate neurotoxic effects. However, the role of astrocytes remains unexplored in these models. Astrocytes have an integral role in the maintenance of central nervous system (CNS) homeostasis, and are associated with both neuroprotection and neurodegradation when they are activated in response to toxic substances or disease states. GFAP is an intermediate filament protein expressed predominantly in the astrocytes of the CNS. Astrocytic activation (gliosis) leads to the upregulation of GFAP, commonly accompanied by astrocyte proliferation and hypertrophy. This process of reactive gliosis has been proposed as an early marker of damage to the nervous system. The traditional method for GFAP quantitation is by immunoassay. This approach is limited by an inability to provide information on cellular localization, morphology and cell number. We determined that HCA could be used to overcome these limitations and to simultaneously measure multiple features associated with gliosis - changes in GFAP expression, astrocyte hypertrophy, and astrocyte proliferation - within a single assay. In co-culture

  7. A neuronal and astrocyte co-culture assay for high content analysis of neurotoxicity.

    Science.gov (United States)

    Anderl, Janet L; Redpath, Stella; Ball, Andrew J

    2009-05-05

    High Content Analysis (HCA) assays combine cells and detection reagents with automated imaging and powerful image analysis algorithms, allowing measurement of multiple cellular phenotypes within a single assay. In this study, we utilized HCA to develop a novel assay for neurotoxicity. Neurotoxicity assessment represents an important part of drug safety evaluation, as well as being a significant focus of environmental protection efforts. Additionally, neurotoxicity is also a well-accepted in vitro marker of the development of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Recently, the application of HCA to neuronal screening has been reported. By labeling neuronal cells with betaIII-tubulin, HCA assays can provide high-throughput, non-subjective, quantitative measurements of parameters such as neuronal number, neurite count and neurite length, all of which can indicate neurotoxic effects. However, the role of astrocytes remains unexplored in these models. Astrocytes have an integral role in the maintenance of central nervous system (CNS) homeostasis, and are associated with both neuroprotection and neurodegradation when they are activated in response to toxic substances or disease states. GFAP is an intermediate filament protein expressed predominantly in the astrocytes of the CNS. Astrocytic activation (gliosis) leads to the upregulation of GFAP, commonly accompanied by astrocyte proliferation and hypertrophy. This process of reactive gliosis has been proposed as an early marker of damage to the nervous system. The traditional method for GFAP quantitation is by immunoassay. This approach is limited by an inability to provide information on cellular localization, morphology and cell number. We determined that HCA could be used to overcome these limitations and to simultaneously measure multiple features associated with gliosis - changes in GFAP expression, astrocyte hypertrophy, and astrocyte proliferation - within a single assay. In co-culture

  8. Skin equivalent tissue-engineered construct: co-cultured fibroblasts/ keratinocytes on 3D matrices of sericin hope cocoons.

    Science.gov (United States)

    Nayak, Sunita; Dey, Sancharika; Kundu, Subhas C

    2013-01-01

    The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from "Sericin Hope" silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-β, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-α, IL-1β and nitric oxide) production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair.

  9. Neural cell co-culture induced differentiation of bone marrow mesenchymal stem cells into neuronal-like cells

    Institute of Scientific and Technical Information of China (English)

    Nailong Yang; Lili Xu; Fen Yang

    2008-01-01

    BACKGROUND: It has been previously demonstrated that the neural cell microenvironment has the ability to induce differentiation of bone marrow mesenchymal stem cells (BMSCs) into the neural cells.OBJECTIVE: To establish a co-culture system of human BMSCs and neural cells, and to observe effects of this co-culture system on differentiation of human BMSCs into neural cells.DESIGN, TIME AND SETTING: A comparative observation experiment, performed at the Center Laboratory of the Affiliated Hospital of Medical College Qingdao University from October 2006 to December 2007.MATERIALS: Neural cells were obtained from human fetal brain tissue. BMSCs were harvested from female patients that underwent autonomous stem cell transplantation.METHODS: BMSCs in the co-culture group consisted of BMSCs and third passage neural cells. BMSCs in the control group were solely cultured in vitro.MAIN OUTCOME MEASURES: Morphological changes of BMSCs were observed, and expression of the neuronal specific marker, neuron-specific enolase (NSE), was analyzed by immunofluorescence staining after4-5-day co-culture.RESULTS: The number of neural cells in the co-culture group increased and the cells spread on the culture bottle surface. Radial dendrite formed and connected with each other. NSE-immunoreactive cells were also detected. The positive ratio of NSE-positive cells reached (32.7±11.5)%, with morphological characteristics similar to neuronal cells. Human BMSCs did not express NSE in the control group.CONCLUSION: The microenvironment provided by neurons induced differentiation of BMSCs into neuronal-like cells.

  10. In vitro co-cultures of Pinus pinaster with Bursaphelenchus xylophilus: a biotechnological approach to study pine wilt disease.

    Science.gov (United States)

    Faria, Jorge M S; Sena, Inês; Vieira da Silva, Inês; Ribeiro, Bruno; Barbosa, Pedro; Ascensão, Lia; Bennett, Richard N; Mota, Manuel; Figueiredo, A Cristina

    2015-06-01

    Co-cultures of Pinus pinaster with Bursaphelenchus xylophilus were established as a biotechnological tool to evaluate the effect of nematotoxics addition in a host/parasite culture system. The pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD), was detected for the first time in Europe in 1999 spreading throughout the pine forests in Portugal and recently in Spain. Plant in vitro cultures may be a useful experimental system to investigate the plant/nematode relationships in loco, thus avoiding the difficulties of field assays. In this study, Pinus pinaster in vitro cultures were established and compared to in vivo 1 year-old plantlets by analyzing shoot structure and volatiles production. In vitro co-cultures were established with the PWN and the effect of the phytoparasite on in vitro shoot structure, water content and volatiles production was evaluated. In vitro shoots showed similar structure and volatiles production to in vivo maritime pine plantlets. The first macroscopic symptoms of PWD were observed about 4 weeks after in vitro co-culture establishment. Nematode population in the culture medium increased and PWNs were detected in gaps of the callus tissue and in cavities developed from the degradation of cambial cells. In terms of volatiles main components, plantlets, P. pinaster cultures, and P. pinaster with B. xylophilus co-cultures were all β- and α-pinene rich. Co-cultures may be an easy-to-handle biotechnological approach to study this pathology, envisioning the understanding of and finding ways to restrain this highly devastating nematode.

  11. Changes of Proliferation and Apoptosis of K562 Cells after Co-culture with Leukemia Bone Marrow Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Katja Karjalainen; Carlos E Bueso-Ramos; Hagop M Kantarjian

    2014-01-01

    Objective:To compare the changes of proliferation and apoptosis of K562 cells after co-culture with human leukemia bone marrow mesenchymal stem cells (LMSC). Methods: The prepared cells were randomly divided into SCG group, SCG+0%FBS group, SCG+0%FBS group and CCG+0%FBS group. Cell counting kit-8 (CCK-8) analytic approach was adopted to detect the optical density (OD) of K562 cells in SCG and CCG groups, and the conditions of K562 cell proliferation under different cultured circumstances were compared. Flow cytometer (FCM) was used to detect the changes of K562 cell cycle after co-culture with LMSC, Annexin V/polyimide (PI) lfuorescence labeling method to detect the changes of K562 cell apoptosis after co-culture with LMSC and serum starvation. Results:After co-culture with LMSC, the proliferation of K562 cells was markedly inhibited, and OD in CCG group was conspicuously lower than that in SCG group. Flow cytometer (FCM) detection on cell cycles demonstrated that after co-culture with LMSC, the proportion of cells in gap phases 0~1 (G0~G1) went up notably, whereas that in phase S went down obviously. Besides, the proportion of cells in phases G2~M was on the rise. K562 cell apoptosis in CCG+0%FBS group was more than in SCG+10%FBS group, and less than in SCG+0%FBS group, indicating LMSC had the function of resisting leukemia cell apoptosis. Conclusion: LMSC exerts the effect of inhibiting the proliferation by blocking K562 cell cycles in phases G0~G1, and inhibiting K562 cell apoptosis induced by serum starvation.

  12. Changes of Proliferation and Apoptosis of K562 Cells after Co-culture with Leukemia Bone Marrow Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Katja Karjalainen

    2014-06-01

    Full Text Available Objective: To compare the changes of proliferation and apoptosis of K562 cells after co-culture with human leukemia bone marrow mesenchymal stem cells (LMSC. Methods: The prepared cells were randomly divided into SCG group, SCG + 0%FBS group, SCG + 0%FBS group and CCG + 0%FBS group. Cell counting kit-8 (CCK-8 analytic approach was adopted to detect the optical density (OD of K562 cells in SCG and CCG groups, and the conditions of K562 cell proliferation under different cultured circumstances were compared. Flow cytometer (FCM was used to detect the changes of K562 cell cycle after co-culture with LMSC, Annexin V/polyimide (PI fluorescence labeling method to detect the changes of K562 cell apoptosis after co-culture with LMSC and serum starvation. Results: After co-culture with LMSC, the proliferation of K562 cells was markedly inhibited, and OD in CCG group was conspicuously lower than that in SCG group. Flow cytometer (FCM detection on cell cycles demonstrated that after co-culture with LMSC, the proportion of cells in gap phases 0 - 1 (G0 - G1 went up notably, whereas that in phase S went down obviously. Besides, the proportion of cells in phases G2 - M was on the rise. K562 cell apoptosis in CCG + 0%FBS group was more than in SCG + 10%FBS group, and less than in SCG + 0%FBS group, indicating LMSC had the function of resisting leukemia cell apoptosis. Conclusion: LMSC exerts the effect of inhibiting the proliferation by blocking K562 cell cycles in phases G0 - G1, and inhibiting K562 cell apoptosis induced by serum starvation.

  13. Astrocytes Enhance Streptococcus suis-Glial Cell Interaction in Primary Astrocyte-Microglial Cell Co-Cultures.

    Science.gov (United States)

    Seele, Jana; Nau, Roland; Prajeeth, Chittappen K; Stangel, Martin; Valentin-Weigand, Peter; Seitz, Maren

    2016-06-13

    Streptococcus (S.) suis infections are the most common cause of meningitis in pigs. Moreover, S. suis is a zoonotic pathogen, which can lead to meningitis in humans, mainly in adults. We assume that glial cells may play a crucial role in host-pathogen interactions during S. suis infection of the central nervous system. Glial cells are considered to possess important functions during inflammation and injury of the brain in bacterial meningitis. In the present study, we established primary astrocyte-microglial cell co-cultures to investigate interactions of S. suis with glial cells. For this purpose, microglial cells and astrocytes were isolated from new-born mouse brains and characterized by flow cytometry, followed by the establishment of astrocyte and microglial cell mono-cultures as well as astrocyte-microglial cell co-cultures. In addition, we prepared microglial cell mono-cultures co-incubated with uninfected astrocyte mono-culture supernatants and astrocyte mono-cultures co-incubated with uninfected microglial cell mono-culture supernatants. After infection of the different cell cultures with S. suis, bacteria-cell association was mainly observed with microglial cells and most prominently with a non-encapsulated mutant of S. suis. A time-dependent induction of NO release was found only in the co-cultures and after co-incubation of microglial cells with uninfected supernatants of astrocyte mono-cultures mainly after infection with the capsular mutant. Only moderate cytotoxic effects were found in co-cultured glial cells after infection with S. suis. Taken together, astrocytes and astrocyte supernatants increased interaction of microglial cells with S. suis. Astrocyte-microglial cell co-cultures are suitable to study S. suis infections and bacteria-cell association as well as NO release by microglial cells was enhanced in the presence of astrocytes.

  14. A bladder cancer microenvironment simulation system based on a microfluidic co-culture model.

    Science.gov (United States)

    Liu, Peng-fei; Cao, Yan-wei; Zhang, Shu-dong; Zhao, Yang; Liu, Xiao-guang; Shi, Hao-qing; Hu, Ke-yao; Zhu, Guan-qun; Ma, Bo; Niu, Hai-tao

    2015-11-10

    A tumor microenvironment may promote tumor metastasis and progression through the dynamic interplay between neoplastic cells and stromal cells. In this work, the most representative and significant stromal cells, fibroblasts, endothelial cells, and macrophages were used as vital component elements and combined with bladder cancer cells to construct a bladder cancer microenvironment simulation system. This is the first report to explore bladder cancer microenvironments based on 4 types of cells co-cultured simultaneously. This simulation system comprises perfusion equipment, matrigel channel units, a medium channel and four indirect contact culture chambers, allowing four types of cells to simultaneously interact through soluble biological factors and metabolites. With this system, bladder cancer cells (T24) with a tendency to form a 'reticular' structure under 3 dimensional culture conditions were observed in real time. The microenvironment characteristics of paracrine interactions and cell motility were successfully simulated in this system. The phenotype change process in stromal cells was successfully reproduced in this system by testing the macrophage effector molecule Arg-1. Arg-1 was highly expressed in the simulated tumor microenvironment group. To develop "precision medicine" in bladder cancer therapy, bladder cancer cells were treated with different clinical 'neo-adjuvant' chemotherapy schemes in this system, and their sensitivity differences were fully reflected. This work provides a preliminary foundation for neo-adjuvant chemotherapy in bladder cancer, a theoretical foundation for tumor microenvironment simulation and promotes individual therapy in bladder cancer patients.

  15. Microalgae Harvest through Fungal Pelletization—Co-Culture of Chlorella vulgaris and Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Sarman Oktovianus Gultom

    2014-07-01

    Full Text Available Microalgae harvesting is a labor- and energy-intensive process and new approaches to harvesting microalgae need to be developed in order to decrease the costs. In this study; co-cultivatation of filamentous fungus (Aspergillus niger and microalgae (Chlorella vulgaris to form cell pellets was evaluated under different conditions, including organic carbon source (glucose; glycerol; and sodium acetate concentration; initial concentration of fungal spores and microalgal cells and light. Results showed that 2 g/L of glucose with a 1:300 ratio of fungi to microalgae provided the best culturing conditions for the process to reach >90% of cell harvest efficiency. The results also showed that an organic carbon source was required to sustain the growth of fungi and form the cell pellets. The microalgae/fungi co-cultures at mixotrophic conditions obtained much higher total biomass than pure cultures of each individual strains; indicating the symbiotic relationship between two strains. This can benefit the microbial biofuel production in terms of cell harvest and biomass production.

  16. Cell-friendly inverse opal-like hydrogels for a spatially separated co-culture system.

    Science.gov (United States)

    Kim, Jaeyun; Bencherif, Sidi A; Li, Weiwei Aileen; Mooney, David J

    2014-09-01

    Three-dimensional macroporous scaffolds have extensively been studied for cell-based tissue engineering but their use is mostly limited to mechanical support for cell adhesion and growth on the surface of macropores. Here, a templated fabrication method is described to prepare cell-friendly inverse opal-like hydrogels (IOHs) allowing both cell encapsulation within the hydrogel matrix and cell seeding on the surface of macropores. Ionically crosslinked alginate microbeads and photocrosslinkable biocompatible polymers are used as a sacrificial template and as a matrix, respectively. The alginate microbeads are easily removed by a chelating agent, with minimal toxicity for the encapsulated cells during template removal. The outer surface of macropores in IOHs can also provide a space for cell adherence. The cells encapsulated or attached in IOHs are able to remain viable and to proliferate over time. The elastic modulus and cell-adhesion properties of IOHs can be easily controlled and tuned. Finally, it is demonstrated that IOH can be used to co-culture two distinct cell populations in different spatial positions. This cell-friendly IOH system provides a 3D scaffold for organizing different cell types in a controllable microenvironment to investigate biological processes such as stem cell niches or tumor microenvironments.

  17. Three-dimensional in vitro co-culture model of breast tumor using magnetic levitation.

    Science.gov (United States)

    Jaganathan, Hamsa; Gage, Jacob; Leonard, Fransisca; Srinivasan, Srimeenakshi; Souza, Glauco R; Dave, Bhuvanesh; Godin, Biana

    2014-10-01

    In this study, we investigate a novel in vitro model to mimic heterogeneous breast tumors without the use of a scaffold while allowing for cell-cell and tumor-fibroblast interactions. Previous studies have shown that magnetic levitation system under conventional culturing conditions results in the formation of three-dimensional (3D) structures, closely resembling in vivo tissues (fat tissue, vasculature, etc.). Three-dimensional heterogeneous tumor models for breast cancer were designed to effectively model the influences of the tumor microenvironment on drug efficiency. Various breast cancer cells were co-cultured with fibroblasts and then magnetically levitated. Size and cell density of the resulting tumors were measured. The model was phenotypically compared to in vivo tumors and examined for the presence of ECM proteins. Lastly, the effects of tumor stroma in the 3D in vitro model on drug transport and efficiency were assessed. Our data suggest that the proposed 3D in vitro breast tumor is advantageous due to the ability to: (1) form large-sized (millimeter in diameter) breast tumor models within 24 h; (2) control tumor cell composition and density; (3) accurately mimic the in vivo tumor microenvironment; and (4) test drug efficiency in an in vitro model that is comparable to in vivo tumors.

  18. Clostridium strain co-cultures for biohydrogen production enhancement from condensed molasses fermentation solubles

    Energy Technology Data Exchange (ETDEWEB)

    Hsiao, Chin-Lang; Wu, Jou-Hsien; Lin, Chiu-Yue [BioHydrogen Lab, Graduate Institute of Civil and Hydraulic Engineering, Feng Chia University, Taichung (China); Chang, Jui-Jen [Genomics Research Center, Academia Sinica, Taipei (China); Department of Life Sciences, National Chung Hsing University, Taichung (China); Chin, Wei-Chih; Wen, Fu-Shyan; Huang, Chieh-Chen [Department of Life Sciences, National Chung Hsing University, Taichung (China); Chen, Chin-Chao [Environmental Resources Laboratory, Department of Landscape Architecture, Chungchou Institute of Technology, Changhwa (China)

    2009-09-15

    An anaerobic continuous-flow hydrogen fermentor was operated at a hydraulic retention time of 8 h using condensed molasses fermentation solubles (CMS) substrate of 40 g-COD/L. Serum bottles were used for seed micro-flora cultivation and batch hydrogen fermentation tests (CMS substrate concentrations of 10-160 g-COD/L). Three hydrogen-producing bacterial strains Clostridium sporosphaeroides F52, Clostridium tyrobutyricum F4 and Clostridium pasteurianum F40 were isolated from the seed fermentor and used as the seeding microbes in single and mixed-culture cultivations for determining their hydrogen productivity. These strains possessed specific hydrogenase genes that could be detected from CMS-fed hydrogen fermentors and were major hydrogen producers. C. pasteurianum F40 was the dominant strain with a high hydrogen production rate while C. sporosphaeroides F52 may play a main role in degrading carbohydrate and glutamate. These strains could be co-cultivated as a symbiotic mixed-culture process to enhance hydrogen productivity. C. pasteurianum F40 or C. tyrobutyricum F4 co-culture with the glutamate-utilizing strain C. sporosphaeroides F52 efficiently enhanced hydrogen production by 12-220% depending on the substrate CMS concentrations. (author)

  19. Immunochemistry of a keratinocyte-fibroblast co-culture model for reconstruction of human skin.

    Science.gov (United States)

    Fleischmajer, R; MacDonald, E D; Contard, P; Perlish, J S

    1993-09-01

    Our purpose was to determine differentiation markers of an in vitro co-culture model in which fibroblasts grown in a three-dimensional nylon mesh were recombined with human keratinocytes. The cultures were kept for 5 weeks and then processed for electron microscopy and immunochemistry. The specimens revealed an epidermis, a basal lamina, an anchoring zone, and a dermis. Epidermal differentiation was confirmed by the presence of K10-keratin, trichohyalin, and filaggrin. The basal lamina contained Type IV collagen, laminin, nidogen, and heparan sulfate. Type IV collagen, laminin, and nidogen were also noted in the extracellular matrix. Type VI collagen was present in the anchoring zone and also gave a reticulated pattern in the rest of the dermis. There was a heavy signal for tenascin and fibronectin throughout the dermis. Osteonectin was restricted to the epidermis and dermal fibroblasts. Fibrillin stained at the anchoring zone and dermis but elastin and vitronectin were negative, suggesting early formation of elastic fibrils. Collagen fibrils stained for Types I, III, and V, as well as the amino propeptide of Types I and III procollagen, suggesting newly synthesized collagen. Decorin was present throughout the dermis. The model described appears suitable for in vitro reconstruction of the skin and may be useful to study the development of various supramolecular skin structures.

  20. Scanning electron microscopical observation of an osteoblast/osteoclast co-culture on micropatterned orthopaedic ceramics

    Directory of Open Access Journals (Sweden)

    Mansur Halai

    2014-09-01

    Full Text Available In biomaterial engineering, the surface of an implant can influence cell differentiation, adhesion and affinity towards the implant. On contact with an implant, bone marrow–derived mesenchymal stromal cells demonstrate differentiation towards bone forming osteoblasts, which can improve osteointegration. The process of micropatterning has been shown to improve osteointegration in polymers, but there are few reports surrounding ceramics. The purpose of this study was to establish a co-culture of bone marrow–derived mesenchymal stromal cells with osteoclast progenitor cells and to observe the response to micropatterned zirconia toughened alumina ceramics with 30 µm diameter pits. The aim was to establish whether the pits were specifically bioactive towards osteogenesis or were generally bioactive and would also stimulate osteoclastogenesis that could potentially lead to osteolysis. We demonstrate specific bioactivity of micropatterns towards osteogenesis, with more nodule formation and less osteoclastogenesis compared to planar controls. In addition, we found that that macrophage and osteoclast-like cells did not interact with the pits and formed fewer full-size osteoclast-like cells on the pitted surfaces. This may have a role when designing ceramic orthopaedic implants.

  1. Indole production provides limited benefit to Escherichia coli during co-culture with Enterococcus faecalis.

    Science.gov (United States)

    Pringle, Shelly L; Palmer, Kelli L; McLean, Robert J C

    2017-01-01

    Escherichia coli lives in the gastrointestinal tract and elsewhere, where it coexists within a mixed population. Indole production enables E. coli to grow with other gram-negative bacteria as indole inhibits N-acyl-homoserine lactone (AHL) quorum regulation. We investigated whether E. coli indole production enhanced competition with gram-positive Enterococcus faecalis, wherein quorum signaling is mediated by small peptides. During planktonic co-culture with E. faecalis, the fitness and population density of E. coli tnaA mutants (unable to produce indole) equaled or surpassed that of E. coli wt. During biofilm growth, the fitness of both populations of E. coli stabilized around 100 %, whereas the fitness of E. faecalis declined over time to 85-90 %, suggesting that biofilm and planktonic populations have different competition strategies. Media supplementation with indole removed the competitive advantage of E. coli tnaA in planktonic populations but enhanced it in biofilm populations. E. coli wt and tnaA showed similar growth in Luria-Bertani (LB) broth. However, E. coli growth was inhibited in the presence of filter-sterilized spent LB from E. faecalis, with inhibition being enhanced by indole. Similarly, there was also an inhibition of E. faecalis growth by proteinaceous components (likely bacteriocins) from spent culture media from both E. coli strains. We conclude that E. coli indole production is not a universal competition strategy, but rather works against gram-negative, AHL-producing bacteria.

  2. A rapid co-culture stamping device for studying intercellular communication

    Science.gov (United States)

    Hassanzadeh-Barforoushi, Amin; Shemesh, Jonathan; Farbehi, Nona; Asadnia, Mohsen; Yeoh, Guan Heng; Harvey, Richard P.; Nordon, Robert E.; Warkiani, Majid Ebrahimi

    2016-10-01

    Regulation of tissue development and repair depends on communication between neighbouring cells. Recent advances in cell micro-contact printing and microfluidics have facilitated the in-vitro study of homotypic and heterotypic cell-cell interaction. Nonetheless, these techniques are still complicated to perform and as a result, are seldom used by biologists. We report here development of a temporarily sealed microfluidic stamping device which utilizes a novel valve design for patterning two adherent cell lines with well-defined interlacing configurations to study cell-cell interactions. We demonstrate post-stamping cell viability of >95%, the stamping of multiple adherent cell types, and the ability to control the seeded cell density. We also show viability, proliferation and migration of cultured cells, enabling analysis of co-culture boundary conditions on cell fate. We also developed an in-vitro model of endothelial and cardiac stem cell interactions, which are thought to regulate coronary repair after myocardial injury. The stamp is fabricated using microfabrication techniques, is operated with a lab pipettor and uses very low reagent volumes of 20 μl with cell injection efficiency of >70%. This easy-to-use device provides a general strategy for micro-patterning of multiple cell types and will be important for studying cell-cell interactions in a multitude of applications.

  3. Effect of co-culture and nutrients supplementation on bioremediation of crude petroleum sludge

    Energy Technology Data Exchange (ETDEWEB)

    Devi, Mamilla Prathima; Reddy, Motakatla Venkateswar; Sarma, Ponnapalli Nageswara; Mohan, Srinivasula Reddy Venkata [Bioengineering and Environmental Centre, Indian Institute of Chemical Technology, Hyderabad (India); Juwarkar, Asha [Environmental Biotechnology Division, National Environmental Engineering Research Institute, Nehru Marg, Nagpur (India)

    2011-10-15

    Ex-situ bioremediation of real-field crude petroleum sludge was evaluated to elucidate the role of co-culture (bioaugmentation) and external nutrients supplementation (biostimulation) under anaerobic microenvironment. Maximum removal of total petroleum hydrocarbons (TPH) was observed by integrating biostimulation with bioaugmentation (R5, 44.01%) followed by bioaugmentation alone (R4, 34.47%), co-substrate supplemented operations [R6, 23.36%; R3, 16.5%; R2, 9.88%] and control (R1, 4.36%). Aromatics fraction showed higher degradation in all the conditions studied. Fate of six selected polycyclic aromatic hydrocarbons (PAHs) was evaluated during bioremediation. Among these, four ring PAHs compounds showed good degradation by integration of biostimulation with bioaugmentation (R5) while bioaugmentation alone (R4) documented good degradation of three ring PAHs. Lower ring PAHs compounds showed good degradation with the application of biostimulation (R6). Fluorescent in situ hybridization (FISH) detected the presence of known PAHs degrading microorganisms viz., Bacillus, Pseudomonas, Acido bacteria, Sulphur reducing bacteria Firmicutes, etc. Application of biostimulation and bioaugmentation strategies alone or in combinations documented noticeable influence on the degradation of petroleum sludge. (Copyright copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  4. Regeneration of human-ear-shaped cartilage by co-culturing human microtia chondrocytes with BMSCs.

    Science.gov (United States)

    Zhang, Lu; He, Aijuan; Yin, Zongqi; Yu, Zheyuan; Luo, Xusong; Liu, Wei; Zhang, Wenjie; Cao, Yilin; Liu, Yu; Zhou, Guangdong

    2014-06-01

    Previously, we had addressed the issues of shape control/maintenance of in vitro engineered human-ear-shaped cartilage. Thus, lack of applicable cell source had become a major concern that blocks clinical translation of this technology. Autologous microtia chondrocytes (MCs) and bone marrow stromal cells (BMSCs) were both promising chondrogenic cells that did not involve obvious donor site morbidity. However, limited cell availability of MCs and ectopic ossification of chondrogenically induced BMSCs in subcutaneous environment greatly restricted their applications in external ear reconstruction. The current study demonstrated that MCs possessed strong proliferation ability but accompanied with rapid loss of chondrogenic ability during passage, indicating a poor feasibility to engineer the entire ear using expanded MCs. Fortunately, the co-transplantation results of MCs and BMSCs (25% MCs and 75% BMSCs) demonstrated a strong chondroinductive ability of MCs to promote stable ectopic chondrogenesis of BMSCs in subcutaneous environment. Moreover, cell labeling demonstrated that BMSCs could transform into chondrocyte-like cells under the chondrogenic niche provided by co-cultured MCs. Most importantly, a human-ear-shaped cartilaginous tissue with delicate structure and proper elasticity was successfully constructed by seeding the mixed cells (MCs and BMSCs) into the pre-shaped biodegradable ear-scaffold followed by 12 weeks of subcutaneous implantation in nude mouse. These results may provide a promising strategy to construct stable ectopic cartilage with MCs and stem cells (BMSCs) for autologous external ear reconstruction.

  5. A comparative study of clinical Aeromonas dhakensis and Aeromonas hydrophila isolates in southern Taiwan: A. dhakensis is more predominant and virulent.

    Science.gov (United States)

    Chen, P-L; Wu, C-J; Chen, C-S; Tsai, P-J; Tang, H-J; Ko, W-C

    2014-07-01

    Aeromonas dhakensis, often phenotypically identified as Aeromonas hydrophila, is an important human pathogen. The present study aimed to compare the clinical and biological features of A. dhakensis and A. hydrophila isolates from human wounds. A total of 80 Aeromonas wound isolates collected between January 2004 and April 2011 were analysed. The species was identified by the DNA sequence matching of rpoD and gyrB (or rpoB if necessary). Most of the Aeromonas isolates were identified as A. dhakensis (37, 46.3%), and 13 (16.3%) as A. hydrophila. Both species alone can cause severe skin and soft-tissue infections. More A. dhakensis isolates were found in wounds exposed to environmental water (32.4% vs 0%, p 0.042). More biofilm formation was noted among A. dhakensis isolates (mean optical density at 570 nm, 1.23 ± 0.09 vs 0.78 ± 0.21, p 0.03). The MICs of ceftriaxone, imipenem and gentamicin for A. dhakensis isolates were higher (p Aeromonas wound isolates, and more virulent than A. hydrophila.

  6. Aeromonas hydrophila meningitis and fulminant sepsis in preterm newborn: A case report and review of literature

    Directory of Open Access Journals (Sweden)

    A Kali

    2016-01-01

    Full Text Available Neonatal meningitis is a lethal infection occurring in the 1st month of life. The risk of developing permanent neurological sequels is high among the neonates who survive. Bacterial pathogens are commonly associated with this condition. Aeromonas is a Gram-negative bacteria of aquatic habitat. Although isolation of Aeromonas species from neonates with blood stream infection is infrequently reported, neonatal meningitis caused by Aeromonas is exceedingly rare. We present a case of fulminant sepsis and meningitis caused by Aeromonas hydrophila in a preterm newborn male. The bacteria was isolated in culture from blood and cerebrospinal fluid. In spite of targeted antibiotics and supportive therapy, the baby failed to respond and died on the 12th day of life.

  7. Draft genome sequences of three Aeromonas hybrophila isolates from catfish and tilapia

    Science.gov (United States)

    Aeromonas hydrophila is a Gram-negative bacteria that is particularly adapted to freshwater environments and can cause severe infections in fish and humans. Here we report the draft genomes of three A. hydrophila catfish and tilapia isolates....

  8. HOST GENE CELL RESEARCH FOR DETERMINING VIRULENCE OF AEROMONAS SPP. COLLECTED FROM ENVIRONMENTAL SAMPLES

    Science.gov (United States)

    The United States Environmental Protection Agency (USEPA) is interested in assessing health risks associated with emerging or potential waterborne pathogens. To this end, the Agency has established a Candidate Contaminant List (CCL) that includes Aeromonas hydrophila an...

  9. IDENTIFICATION AND CHARACTERIZATION OF AEROMONAS ISOLATES FROM DRINKING WATER DISTRIBUTION SYSTEMS

    Science.gov (United States)

    Members of the bacterial genus Aeromonas are commonly isolated from both fresh and salt waters worldwide and some are believed to cause infections in humans, including gastroenteritis and wound infections. Currently, aeromonads are on the United States Environmental Protection A...

  10. Glia and epilepsy: experimental investigation of antiepileptic drugs in an astroglia/microglia co-culture model of inflammation.

    Science.gov (United States)

    Dambach, Hannes; Hinkerohe, Daniel; Prochnow, Nora; Stienen, Martin N; Moinfar, Zahra; Haase, Claus G; Hufnagel, Andreas; Faustmann, Pedro M

    2014-01-01

    The contribution of glial cells, mainly astrocytes and microglia, to the pathophysiology of epilepsy is increasingly appreciated. Glia play a pivotal role in the initiation and maintenance of the central nervous system (CNS) immune response and neuronal metabolic and trophic supply. Recent clinical and experimental evidence suggests a direct relationship between epileptic activity and CNS inflammation, which is characterized by accumulation, activation, and proliferation of microglia and astrocytes. Concomitant glia-mediated mechanisms of action of several antiepileptic drugs (AEDs) have been proposed. However, their direct effects on glial cells have been rarely investigated. We aimed to investigate the effect of commonly used AEDs on glial viability, the gap junctional network, the microglial activation, and cytokine expression in an in vitro astroglia/microglia co-culture model. Primary astrocytic cultures were prepared from brains of postnatal (P0-P2) Wistar rats and co-cultured with a physiologic amount of 5%, as well as 30% microglia in order to mimic inflammatory conditions. Co-cultures were treated with valproic acid (VPA), carbamazepine (CBZ), phenytoin (PHE), and gabapentin (GBT). Viability and proliferation were measured using the tetrazolium (MTT) assay. The microglial activation state was determined by immunocytochemical labeling. The astroglial connexin 43 (Cx43) expression was measured by Western blot analysis. The transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) cytokine levels were measured by the quantitative sandwich enzyme immunosorbent assay (ELISA). Astrocytes, co-cultured with 5% microglia (M5 co-cultures), showed a dose-dependent, significant reduction in glial viability after incubation with PHE and CBZ. Furthermore, VPA led to highly significant microglial activation at all doses examined. The antiinflammatory cytokine TGF-β1 release was induced by high doses of GBT and PHE. Astrocytes co-cultured with 30

  11. Anti-biogram and resistogram profiling of Aeromonas species isolated from Malaysian aquatic sources

    OpenAIRE

    Olumide Adedokun Odeyemi; Asmat Ahmad

    2013-01-01

    Objective: To investigate antibiotics and heavy metals resistance profile of Aeromonas species isolated from Kuala Gula and Kuala Sepetang, Perak. Methods: Isolated Aeromonas species were subjected to 12 antibiotics: penicillin G 10 units, tetracycline 30 µg, novobiocin 5 µg, nitrofurantoin 100 µg, ampicillin 10 µg, chloramphenicol 10 µg, fluconazole 25 µg, gentamicin 10 µg, streptomycin 10 µg and 25 µg, kanamycin 30 µg, and vancomycin 30 µg and 5 heavy metals: MnSO4.H2O, Cr(NO...

  12. Fibroblasts weaken the anti-tumor effect of gefitinib on co-cultured non-small cell lung cancer cells

    Institute of Scientific and Technical Information of China (English)

    Yong Xiao; Wang Peiqin; Jiang Tao; Yu Wenchen; Shang Yan; Han Yiping; Zhang Pingping

    2014-01-01

    Background Non-small cell lung cancer (NSCLC) is the most common lung malignancy worldwide.The metastatic potential of NSCLC cells has been shown to be associated with the tumor microenvironment,which consists of tumor cells,stroma,blood vessels,immune infiltrates and the extracellular matrix.Fibroblasts can produce numerous extraceilular matrix molecules and growth factors.Gefitinib has been evaluated as a first-line treatment in selected patients,and it has shown favorable efficacy especially in NSCLC,but it is not effective for everyone.Methods In this study,we examined the antitumor activity of gefitinib on lung fibroblasts co-cultured of lung cancer cells.A series of co-culture experiments that employed cell counting kit-8 (CCK8),transwells,real-time polymerase chain reaction (RT-PCR) and Western blotting with HFL-1 fibroblasts and A549 human lung carcinoma cells were performed to learn more about tumor cell proliferation,migration and invasion; and to determine any change of epithelial mesenchymal transition (EMT)-associated tumor markers vimentin,matrix metallopro-teinase 2 (MMP2) and chemotaxis cytokines receptor 4 (CXCR4) mRNA levels.Results A549 cell proliferation in the presence of HFL-1 cells was not significantly increased compared with A549 cells alone,but A549 cell spheroid body formation was increased after co-culture,and treatment with gefitinib increased further.Our study also revealed that fibroblasts attenuated the lung cancer cell inhibition ratio of migration and invasion after gefitinib treatment in vitro.To further study this mechanism,RT-PCR analysis showed that vimentin,MMP2 and CXCR4 mRNA levels were more highly expressed in the lung cancer cells after co-culture,but did not obviously decrease compared with the control cells following gefitinib treatment.This suggests the mechanism by which fibroblasts attenuate gefitinib-induced expression of EMT-associated tumor markers.Finally,our results demonstrated that co-culture with A549 lung

  13. Isolation and Seroprevalence of Aeromonas spp. Among Common Food Animals Slaughtered in Nagpur, Central India.

    Science.gov (United States)

    Gowda, Tanuja K G M; Reddy, Vishwanatha R A P; Devleesschauwer, Brecht; Zade, Nandkishor N; Chaudhari, Sandeep P; Khan, Waqar A; Shinde, Shilpa V; Patil, Archana R

    2015-07-01

    Aeromonads are ubiquitous foodborne pathogens with a global distribution. Animal-origin foods and contaminated animals are the main sources of Aeromonas infection to humans. So far little is known about the occurrence of Aeromonas spp. in food-producing animals in India. The present study was conducted to determine the prevalence and seroprevalence of Aeromonas species from 50 each of meat, blood, and sera samples collected from cattle, buffaloes, goats, and pigs slaughtered in and around Nagpur, Central India. Alkaline peptone water and ampicillin dextrin agar were used to isolate Aeromonas spp. An indirect enzyme-linked immunosorbent assay (ELISA) was standardized by use of whole-cell antigen (WC) and outer membrane protein (OMP) of Aeromonas hydrophila (MTCC 646). Aeromonads were isolated from 44 (22%) of the meat samples, and 1 (0.5%) from the blood samples. Seroprevalence by indirect ELISA-based WC antigen was estimated as 68% in cattle, 44% in buffaloes, 60% in goats, and 30% in pigs. OMP-based ELISA yielded a seroprevalence of 56%, 48%, 52%, and 22% in cattle, buffaloes, goats, and pigs, respectively. The results revealed that OMP-based ELISA and WC-based ELISA were in agreement with one another. Isolation along with high seropositivity demonstrates the presence of foodborne Aeromonas spp. in the Nagpur city of Central India.

  14. High frequency of coinfecting enteropathogens in Aeromonas-associated diarrhea of hospitalized Peruvian infants.

    Science.gov (United States)

    Pazzaglia, G; Sack, R B; Salazar, E; Yi, A; Chea, E; Leon-Barua, R; Guerrero, C E; Palomino, J

    1991-06-01

    Rectal swabs from 391 infants less than 18 months of age who were hospitalized with acute diarrhea and from 138 similarly aged healthy infants were examined for the etiologic agents of diarrhea. Aeromonas spp. were recovered from 205 of 391 (52.4%) diarrheic patients, whereas they were recovered from 12 of 138 (8.7%) controls (P less than 10(-11). Among the 205 Aeromonas-positive diarrheic patients, 118 (57.6%) were found to be coinfected with other common enteropathogens. Of the 164 Aeromonas-positive initial diarrheic specimens, 82 (50.0%) had one or more other enteropathogens present; 30 patients were coinfected with rotavirus, 20 with enterotoxigenic Escherichia coli, 16 with Campylobacter spp., 14 with Shigella spp., 13 with enteropathogenic E. coli, 4 with Vibrio spp., 1 with Salmonella spp., and 1 with Plesiomonas spp. of Aeromonas strains from cases compared with that from controls supports an etiologic role for this organism. However, frequent concomitant infections with other well-recognized enteropathogens and a lack of disease correlation with common Aeromonas phenotypes suggest that only a subset of Aeromonas strains may be diarrhea causing and that such strains may be common to several of the existing species.

  15. Aeromonas hydrophila Septicemia in Acute Hand Injury: A Case Report

    Directory of Open Access Journals (Sweden)

    Chin-Chiang Yang

    2004-07-01

    Full Text Available Aeromonas hydrophila is an uncommon and frequently overlooked cause of skin and soft-tissue infection. Rapid onset of cellulitis and bullae formation in the setting of soft-tissue trauma in connection with exposure to fresh water should alert the clinician to the possibility of infection by this organism. We report a case of severe necrotizing fasciitis due to A. hydrophila infection involving the hand and arm, complicated by septicemia within 30 hours of injury. In addition to appropriate antibiotic therapy and general supportive care, treatment involves early surgical exploration for compartment decompression and aggressive debridement of all necrotic tissue followed by skin grafting for delayed wound closure. There was good functional outcome after more than 1 year of follow-up. Early awareness of A. hydrophila infection, as well as of infection by Vibrio, is necessary in preventing crippling deformities of the hand in post-traumatic wound infection with a history of exposure to fresh water.

  16. Isolation of Aeromonas hydrophila from the American alligator, Alligator mississippiensis

    Energy Technology Data Exchange (ETDEWEB)

    Gorden, R.W. (Univ. of Southern Colorado, Pueblo); Hazen, T.C.; Esch, G.W.; Fliermans, C.B.

    1979-04-01

    Aeromonas hydrophila was isolated from the internal organs of nine adult alligators, Alligator mississippiensis, which died without apparent cause, suggesting the bacterium may have been a factor. One hundred and twenty-three alligators ranging in age from six months to over 10 years were captured from five locations in the southeastern United States and sampled for A. hydrophila. The bacterium was isolated from the oral cavity of 85% of the animals, on the external jaw area from over 50% and from 70% of the internal tissue samples. A. hydrophila is ubiquitous with alligators in their natural habitats, but apparently does not cause clinical disease. However, stress factors such as trapping, handling, and warm water tempertures may be conducive to the rapid proliferation of the bacteria, thereby facilitating disease.

  17. Mortality of therapeutic fish Garra rufa caused by Aeromonas sobria

    Institute of Scientific and Technical Information of China (English)

    Juraj Majtn; Jaroslaverny; Alena Ofkan; Peter Tak; Milan Koznek

    2012-01-01

    Objective:To investigate a case of mass mortality of Garra rufa (G. rufa) from a fish hatchery farm in Slovakia. Methods: Causative bacterial agent was swabbing out of affected fish skin area and subsequently identified using commercial test system. Antibiotic susceptibility was determined by the disk diffusion method. Results: Infected G. rufa was characterized by abnormal swimming behaviour, bleeding of skin lesions and local haemorrhages. Despite of using recommended aquatic antibiotic treatment no improvement was achieved and Aeromonas sobria (A. sobria) was identified as a causative agent of fish mortality. Due to massive fish mortality, antibiotic susceptibility of pure isolated culture of A. sobria was evaluated employing eight antibiotics against human infections. A. sobria was resistant only against one antibiotic, namely ampicilin. Conclusions: These results indicate that A. sobria can act as a primary pathogen of G. rufa and may be a potential risk factor for immunodeficient or immunoincompetent patients during the ichthyotherapy.

  18. Astroglia overexpressing heme oxygenase-1 predispose co-cultured PC12 cells to oxidative injury.

    Science.gov (United States)

    Song, Linyang; Song, Wei; Schipper, Hyman M

    2007-08-01

    The mechanisms responsible for the progressive degeneration of dopaminergic neurons and pathologic iron deposition in the substantia nigra pars compacta of patients with Parkinson's disease (PD) remain unclear. Heme oxygenase-1 (HO-1), the rate-limiting enzyme in the oxidative degradation of heme to ferrous iron, carbon monoxide, and biliverdin, is upregulated in affected PD astroglia and may contribute to abnormal mitochondrial iron sequestration in these cells. To determine whether glial HO-1 hyper-expression is toxic to neuronal compartments, we co-cultured dopaminergic PC12 cells atop monolayers of human (h) HO-1 transfected, sham-transfected, or non-transfected primary rat astroglia. We observed that PC12 cells grown atop hHO-1 transfected astrocytes, but not the astroglia themselves, were significantly more susceptible to dopamine (1 microM) + H(2)O(2) (1 microM)-induced death (assessed by nuclear ethidium monoazide bromide staining and anti-tyrosine hydroxylase immunofluorescence microscopy) relative to control preparations. In the experimental group, PC12 cell death was attenuated significantly by the administration of the HO inhibitor, SnMP (1.5 microM), the antioxidant, ascorbate (200 microM), or the iron chelators, deferoxamine (400 microM), and phenanthroline (100 microM). Exposure to conditioned media derived from HO-1 transfected astrocytes also augmented PC12 cell killing in response to dopamine (1 microM) + H(2)O(2) (1 microM) relative to control media. In PD brain, overexpression of HO-1 in nigral astroglia and accompanying iron liberation may facilitate the bioactivation of dopamine to neurotoxic free radical intermediates and predispose nearby neuronal constituents to oxidative damage. (c) 2007 Wiley-Liss, Inc.

  19. Mesenchymal stem cells enhance GABAergic transmission in co-cultured hippocampal neurons.

    Science.gov (United States)

    Mauri, Mario; Lentini, Daniela; Gravati, Marta; Foudah, Dana; Biella, Gerardo; Costa, Barbara; Toselli, Mauro; Parenti, Marco; Coco, Silvia

    2012-04-01

    Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent stem cells endowed with neurotrophic potential combined with immunological properties, making them a promising therapeutic tool for neurodegenerative disorders. However, the mechanisms through which MSCs promote the neurological recovery following injury or inflammation are still largely unknown, although cell replacement and paracrine mechanisms have been hypothesized. In order to find out what are the mechanisms of the trophic action of MSCs, as compared to glial cells, on CNS neurons, we set up a co-culture system where rat MSCs (or cortical astrocytes) were used as a feeding layer for hippocampal neurons without any direct contact between the two cell types. The analysis of hippocampal synaptogenesis, synaptic vesicle recycling and electrical activity show that MSCs were capable to support morphological and functional neuronal differentiation. The proliferation of hippocampal glial cells induced by the release of bioactive substance(s) from MSCs was necessary for neuronal survival. Furthermore, MSCs selectively increased hippocampal GABAergic pre-synapses. This effect was paralleled with a higher expression of the potassium/chloride KCC2 co-transporter and increased frequency and amplitude of mIPSCs and sIPSCs. The enhancement of GABA synapses was impaired by the treatment with K252a, a Trk/neurotrophin receptor blocker, and by TrkB receptor bodies hence suggesting the involvement of BDNF as a mediator of such effects. The results obtained here indicate that MSC-secreted factors induce glial-dependent neuronal survival and trigger an augmented GABAergic transmission in hippocampal cultures, highlighting a new effect by which MSCs could promote CNS repair. Our results suggest that MSCs may be useful in those neurological disorders characterized by an impairment of excitation versus inhibition balance.

  20. Irradiation Can Selectively Kill Tumor Cells while Preserving Erythrocyte Viability in a Co-Culture System.

    Directory of Open Access Journals (Sweden)

    Ming Gong

    Full Text Available An understanding of how to safely apply intraoperative blood salvage (IBS in cancer surgery has not yet been obtained. Here, we investigated the optimal dose of 137Cs gamma-ray irradiation for killing human hepatocarcinoma (HepG2, gastrocarcinoma (SGC7901, and colonic carcinoma (SW620 tumor cells while preserving co-cultured erythrocytes obtained from 14 healthy adult volunteers. HepG2, SGC7901, or SW620 cells were mixed into the aliquots of erythrocytes. After the mixed cells were treated with 137Cs gamma-ray irradiation (30, 50, and 100 Gy, tumor cells and erythrocytes were separated by density gradient centrifugation in Percoll with a density of 1.063 g/ml. The viability, clonogenicity, DNA synthesis, tumorigenicity, and apoptosis of the tumor cells were determined by MTT assay, plate colony formation, 5-ethynyl-2'-deoxyuridine (EdU incorporation, subcutaneous xenograft implantation into immunocompromised mice, and annexin V/7-AAD staining, respectively. The ATP concentration, 2,3-DPG level, free Hb concentration, osmotic fragility, membrane phosphatidylserine externalization, blood gas variables, reactive oxygen species levels, and superoxide dismutase levels in erythrocytes were analyzed. We found that 137Cs gamma-ray irradiation at 50 Gy effectively inhibited the viability, proliferation, and tumorigenicity of HepG2, SGC7901, and SW620 cells without markedly damaging the oxygen-carrying ability or membrane integrity or increasing the oxidative stress of erythrocytes in vitro. These results demonstrated that 50 Gy irradiation in a standard 137Cs blood irradiator might be a safe and effective method of inactivating HepG2, SGC7901, and SW620 cells mixed with erythrocytes, which might help to safely allow IBS in cancer surgery.

  1. Construction of an in vitro primary lung co-culture platform derived from New Zealand white rabbits

    Energy Technology Data Exchange (ETDEWEB)

    Powell, Joshua D.; Hess, Becky M.; Hutchison, Janine R.; Straub, Tim M.

    2015-05-01

    We report the construction of an in vitro three dimensional (3D) co-culture platform consisting of differentiated lung epithelial cells and monocytes from New Zealand white rabbits. Rabbit lung epithelial cells were successfully grown at air-liquid interface, produced mucus, and expressed both sialic acid alpha-2,3 and alpha-2,6. Blood-derived CD14+ monocytes were deposited above the epithelial layer resulting in the differentiation of a subset of monocytes into CD11c+ cells within the co-culture. These proof-of-concept findings provide a convenient means to comparatively study in vitro versus in vivo rabbit lung responses as they relate to inhalation or lung-challenge studies.

  2. The use of 90% Aloe vera freeze drying as the modulator of collagen density in extraction socket of incicivus Cavia cobaya

    Directory of Open Access Journals (Sweden)

    Ester Arijani

    2008-06-01

    Full Text Available Wound healing is basically a complex process in which cellular and matrix act in concern to re-establish the integrity of injury tissues. This process can be simplified to be healing process consisted of haemostatic, inflammation, cell proliferation and tissue remodeling. The aimed of this research was to know the influence of freeze drying 90% Aloe vera application as mandible collagen density modulator in extraction socket of incisive Cavia cobaya. This research was done using Post Test Only Control Groups Design and Cavia cobaya as the sample. Six samples of each control group and 90% Aloe vera group applied to test each collagen density for three days and seven days. Then, the data was analyzed statistically using Mann Whitney with 5% significance rate. The result of the study indicates that administering 90% Aloe vera can accelerate the growth of collagen density in healing process of extraction socket. The conclusion is 90% Aloe vera can modulate the density of collageneous fiber in socket of extraction incicivus tooth wound of Cavia cobaya.

  3. STUDIES ON THE BACTERIOPHAGE OF D'HERELLE : VI. ON THE VIRULENCE OF THE OVERGROWTH IN THE LYSED CULTURES OF BACILLUS PESTIS CAVIAE (M. T. II).

    Science.gov (United States)

    Bronfenbrenner, J; Muckenfuss, R S; Korb, C

    1926-10-31

    Resistants isolated from the overgrowth of cultures of B. pestis caviae (M. T. II) lysed by various strains of specific bacteriophage proved to be avirulent when administered to mice by feeding, or by intraperitoneal injection. These cultures remained resistant to the action of bacteriophage so long as they were carried on agar. When transferred to broth, however, one group of resistants, namely, those isolated by means of "weak" phages, became susceptible to lysis after five to seven daily passages. The other group of resistants, isolated from the cultures lysed by one of the "strong" phages, failed to become susceptible to lysis even after nearly 200 passages in broth. Simultaneously with the recovery of susceptibility, the cultures of the first group regained a degree of virulence comparable to that of the parent culture of B. pestis caviae. The cultures of the second group of resistants have failed thus far to recover virulence (10 months after isolation). The latter cultures, apart from lack of both virulence and susceptibility to lysis, are identical with the parent culture of B. pestis caviae, as indicated by biochemical and antigenic properties. Our findings offer evidence in favor of the view that resistant strains result from selection among variants already existing in the parent culture and do not arise through the inheritance of specific immunity properties produced by the action of phage.

  4. Oxidative damage to DNA by diesel exhaust particle exposure in co-cultures of human lung epithelial cells and macrophages

    DEFF Research Database (Denmark)

    Jantzen, Kim; Roursgaard, Martin; Madsen, Claus Desler

    2012-01-01

    -DNA glycosylase or oxoguanine DNA glycosylase (hOGG1) sensitive sites, in mono-cultures of A549 or THP-1a and co-cultures of A549 and THP-1a cells. The strongest genotoxic effects were observed in A549 mono-cultures and SRM2975 was more potent than SRM1650b. The ROS production only increased in cells exposed...

  5. Co-culture of vascular endothelial cells and smooth muscle cells by hyaluronic acid micro-pattern on titanium surface

    Science.gov (United States)

    Li, Jingan; Li, Guicai; Zhang, Kun; Liao, Yuzhen; Yang, Ping; Maitz, Manfred F.; Huang, Nan

    2013-05-01

    Micro-patterning as an effective bio-modification technique is increasingly used in the development of biomaterials with superior mechanical and biological properties. However, as of now, little is known about the simultaneous regulation of endothelial cells (EC) and smooth muscle cells (SMC) by cardiovascular implants. In this study, a co-culture system of EC and SMC was built on titanium surface by the high molecular weight hyaluronic acid (HMW-HA) micro-pattern. Firstly, the micro-pattern sample with a geometry of 25 μm wide HMW-HA ridges, and 25 μm alkali-activated Ti grooves was prepared by microtransfer molding (μTM) for regulating SMC morphology. Secondly, hyaluronidase was used to decompose high molecular weight hyaluronic acid into low molecular weight hyaluronic acid which could promote EC adhesion. Finally, the morphology of the adherent EC was elongated by the SMC micro-pattern. The surface morphology of the patterned Ti was imaged by SEM. The existence of high molecular weight hyaluronic acid on the modified Ti surface was demonstrated by FTIR. The SMC micro-pattern and EC/SMC co-culture system were characterized by immunofluorescence microscopy. The nitric oxide release test and cell retention calculation were used to evaluate EC function on inhibiting hyperplasia and cell shedding, respectively. The results indicate that EC in EC/SMC co-culture system displayed a higher NO release and cell retention compared with EC cultured alone. It can be suggested that the EC/SMC co-culture system possessed superiority to EC cultured alone in inhibiting hyperplasia and cell shedding at least in a short time of 24 h.

  6. Development of melanocye-keratinocyte co-culture model for controls and vitiligo to assess regulators of pigmentation and melanocytes

    OpenAIRE

    2012-01-01

    Background: There is a need to develop an in vitro skin models which can be used as alternative system for research and testing pharmacological products in place of laboratory animals. Therefore to study the biology and pathophysiology of pigmentation and vitiligo, reliable in vitro skin pigmentation models are required. Aim: In this study, we used primary cultured melanocytes and keratinocytes to prepare the skin co-culture model in control and vitiligo patients. Methods: The skin grafts wer...

  7. Selective sensitiveness of mesenchymal stem cells to shock waves leads to anticancer effect in human cancer cell co-cultures.

    Science.gov (United States)

    Foglietta, Federica; Duchi, Serena; Canaparo, Roberto; Varchi, Greta; Lucarelli, Enrico; Dozza, Barbara; Serpe, Loredana

    2017-03-15

    Mesenchymal stem cells (MSC) possess the distinctive feature of homing in on and engrafting into the tumor stroma making their therapeutic applications in cancer treatment very promising. Research into new effectors and external stimuli, which can selectively trigger the release of cytotoxic species from MSC toward the cancer cells, significantly raises their potential. Shock waves (SW) have recently gained recognition for their ability to induce specific biological effects, such as the local generation of cytotoxic reactive oxygen species (ROS) in a non-invasive and tunable manner. We thus investigate whether MSC are able to generate ROS and, in turn, affect cancer cell growth when in co-culture with human glioblastoma (U87) or osteosarcoma (U2OS) cells and exposed to SW. MSC were found to be the cell line that was most sensitive to SW treatment as shown by SW-induced ROS production and cytotoxicity. Notably, U87 and U2OS cancer cell growth was unaffected by SW exposure. However, significant decreases in cancer cell growth, 1.8 fold for U87 and 2.3 fold for U2OS, were observed 24h after the SW treatment of MSC co-cultures with cancer cells. The ROS production induced in MSC by SW exposure was then responsible for lipid peroxidation and cell death in U87 and U2OS cells co-cultured with MSC. This experiment highlights the unique ability of MSC to generate ROS upon SW treatment and induce the cell death of co-cultured cancer cells. SW might therefore be proposed as an innovative tool for MSC-mediated cancer treatment. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Optimization of Chlorella vulgaris and bioflocculant-producing bacteria co-culture: enhancing microalgae harvesting and lipid content.

    Science.gov (United States)

    Wang, Y; Yang, Y; Ma, F; Xuan, L; Xu, Y; Huo, H; Zhou, D; Dong, S

    2015-05-01

    Microalgae are a sustainable bioresource, and the biofuel they produce is widely considered to be an alternative to limited natural fuel resources. However, microalgae harvesting is a bottleneck in the development of technology. Axenic Chlorella vulgaris microalgae exhibit poor harvesting, as expressed by a flocculation efficiency of 0·2%. This work optimized the co-culture conditions of C. vulgaris and bioflocculant-producing bacteria in synthetic wastewater using response surface methodology (RSM), thus aiming to enhance C. vulgaris harvesting and lipid content. Three significant process variables- inoculation ratio of bacteria and microalgae, initial glucose concentration, and co-culture time- were proposed in the RSM model. F-values (3·98/8·46) and R(2) values (0·7817/0·8711) both indicated a reasonable prediction by the RSM model. The results showed that C. vulgaris harvesting efficiency reached 45·0-50·0%, and the lipid content was over 21·0% when co-cultured with bioflocculant-producing bacteria under the optimized culture conditions of inoculation ratio of bacteria and microalgae of 0·20-0·25, initial glucose concentration of microalgae harvesting and cost-effective microalgal bioproducts, and confirmed the promising prospect of introducing bioflocculant-producing bacteria into microalgae bioenergy production. This work optimized the co-culture conditions of microalgae (C. vulgaris) and bioflocculant-producing bacteria (F2, Rhizobium radiobacter) in synthetic wastewater using response surface methodology, aiming to enhance C. vulgaris harvesting and lipid produced content. Bioflocculant-producing microbes are environmentally friendly functional materials. They avoid the negative effects of traditional chemical flocculants. This work provided new insights into microalgae harvesting and cost-effective production of microalgal bioproducts, and confirmed the promising prospect of introducing bioflocculant-producing bacteria into microalgae

  9. Retinal pigment epithelial cells upregulate expression of complement factors after co-culture with activated T cells

    DEFF Research Database (Denmark)

    Juel, Helene Bæk; Kaestel, Charlotte; Folkersen, Lasse

    2011-01-01

    In this study we examined the effect of T cell-derived cytokines on retinal pigment epithelial (RPE) cells with respect to expression of complement components. We used an in vitro co-culture system in which CD3/CD28-activated human T cells were separated from the human RPE cell line (ARPE-19) by ...... of inflammatory ocular diseases such as uveitis and age-related macular degeneration. --------------------------------------------------------------------------------...

  10. Stem and Progenitor Cell Expansion in Co-culture of Mobilized CD34 + Cells and Osteopetrotic Mouse Stroma

    Institute of Scientific and Technical Information of China (English)

    Na LI; Shahin Rafii; JF Stoltz; Malcolm A.S. Moore; Pierre Feugier; Deog-Yeon JO; Jae Hung Shieh; Karen L. MacKenzie; JF Lesesve; V Latger-Cannard; D Bensoussan; Ronald G Crystal

    2005-01-01

    @@ 1 Introduction Culture systems capable of expanding and/or maintaining hematopoietic stem cells will not only facilitate our understanding of stem cell biology, but also broaden clinical applications. Among various in vitro hematopoietic culture systems, co-cultures of marrow or CD34+ cells with an adherent stromal layer that can produce cytokines and extracellular matrix components most effectively supports long-term hematopoiesis ( LTC ), mimicking the bone marrow micro-environment.

  11. Fluorescence-based co-culture of normal and cancerous cells as an indicator of therapeutic effects in cancer.

    Science.gov (United States)

    Tamura, Masato; Matsui, Hirofumi; Hyodo, Ichinosuke; Tanaka, Junko; Miwa, Yoshihiro

    2014-10-15

    Comprehensive evaluation of the effects of cancer therapies in vitro is difficult because of the need to distinguish the main effects from the side effects within the data. This problem cannot be overcome by methods involving monoculture, because the effects of anti-cancer drugs in a monoculture can only be measured on either normal or cancerous cells in isolation. In order to promote therapeutic development, therefore, we need a novel drug evaluation method which can simultaneously determine both therapeutic activity and toxicity under a co-culture of normal and cancerous cells. Co-culture creates a more biomimetic condition in comparison to monoculture. The novel method proposed in this study uses an easy experiment for estimating the effects of treatments with various kinds of drugs as a solution to the abovementioned problems. We have previously established two cell lines: a rat gastric mucosal cell line (RGM) and its corresponding cancerous mutant cell line (RGK). In this study, we have developed a new evaluation procedure using a co-culture of green fluorescent protein-expressing RGM cells (RGM-GFP) and kusabira orange-expressing RGK cells (RGK-KO). These cell lines emit green and red fluorescence, respectively. We demonstrated the capability of the method in evaluations of the cancer-selective effects of anti-cancer drugs and X-ray treatment. These results clearly distinguished the cancer-selective toxicity of the applied therapies.

  12. [Effects of Ulva pertusa and Gracilaria lemaneiformis on growth of Heterosigma akashiwo (Raphidophyceae) in co-culture].

    Science.gov (United States)

    Wang, You; Yu, Zhi-ming; Song, Xiu-xian; Zhang, Shan-dong

    2006-02-01

    We studied the effects of fresh tissue and culture medium filtrate of two species of macroalgae, Ulva pertusa (Chlorophyta) and Gracilaria lemaneiformis (Rhodophyta) on growth of Heterosigma akashiwo (Raphidophyceae) in co-culture. Both U. pertusa and G. lemaneiformis, and especially their fresh tissues, significantly impede the growth of H. akashiwo. Carbonate limitations and the presence of environment bacteria are not necessary for the negative effects of macroalgal on H. akashiwo. The simultaneous nutrient assays show that nitrate and phosphate are almost exhausted in the G. lemaneiformis co-culture system, but remain at acceptable levels in the U. pertusa system, when all cells of H. akashiwo are completely dead. When f/2 medium is supplied daily to G. lemaneiformis culture, the growth of H. akashiwo is greatly inhibited but not completely terminated. Furthermore, different amounts of fresh seaweed tissue, and culture medium filtrate prepared from different macroalgal concentrations are analyzed to determine their effects on the growth of H. akashiwo. The results show a positive correlation between the initial macroalgal concentration and the negative effects they exert on the co-cultured microalgae. Results suggest that the allelopathic effects of U. pertusa may be essential for negative effects on H. akashiwo; however, the combined roles of allelopathy and nutrient competition may be responsible for the negative effect of G. lemaneiformis the release of allelochemicals by U. pertusa.

  13. A primary human macrophage-enteroid co-culture model to investigate mucosal gut physiology and host-pathogen interactions

    Science.gov (United States)

    Noel, Gaelle; Baetz, Nicholas W.; Staab, Janet F.; Donowitz, Mark; Kovbasnjuk, Olga; Pasetti, Marcela F.; Zachos, Nicholas C.

    2017-01-01

    Integration of the intestinal epithelium and the mucosal immune system is critical for gut homeostasis. The intestinal epithelium is a functional barrier that secludes luminal content, senses changes in the gut microenvironment, and releases immune regulators that signal underlying immune cells. However, interactions between epithelial and innate immune cells to maintain barrier integrity and prevent infection are complex and poorly understood. We developed and characterized a primary human macrophage-enteroid co-culture model for in-depth studies of epithelial and macrophage interactions. Human intestinal stem cell-derived enteroid monolayers co-cultured with human monocyte-derived macrophages were used to evaluate barrier function, cytokine secretion, and protein expression under basal conditions and following bacterial infection. Macrophages enhanced barrier function and maturity of enteroid monolayers as indicated by increased transepithelial electrical resistance and cell height. Communication between the epithelium and macrophages was demonstrated through morphological changes and cytokine production. Intraepithelial macrophage projections, efficient phagocytosis, and stabilized enteroid barrier function revealed a coordinated response to enterotoxigenic and enteropathogenic E. coli infections. In summary, we have established the first primary human macrophage-enteroid co-culture system, defined conditions that allow for a practical and reproducible culture model, and demonstrated its suitability to study gut physiology and host responses to enteric pathogens. PMID:28345602

  14. A co-culture device with a tunable stiffness to understand combinatorial cell-cell and cell-matrix interactions

    Science.gov (United States)

    Rao, Nikhil; Grover, Gregory N.; Vincent, Ludovic G.; Evans, Samantha C.; Choi, Yu Suk; Vincent, Ludovic G.; Spencer, Katrina H.; Hui, Elliot E.; Engler, Adam J.; Christman, Karen L.

    2013-01-01

    Cell behavior on 2-D in vitro cultures is continually being improved to better mimic in vivo physiological conditions by combining niche cues including multiple cell types and substrate stiffness, which are well known to impact cell phenotype. However, no system exists in which a user can systematically examine cell behavior on a substrate with a specific stiffness (elastic modulus) in culture with a different cell type, while maintaining distinct cell populations. We demonstrate the modification of a silicon reconfigurable co-culture system with a covalently linked hydrogel of user-defined stiffness. This device allows the user to control whether two separate cell populations are in contact with each other or only experience paracrine interactions on substrates of controllable stiffness. To illustrate the utility of this device, we examined the role of substrate stiffness combined with myoblast co-culture on adipose derived stem cell (ASC) differentiation and found that the presence of myoblasts and a 10 kPa substrate stiffness increased ASC myogenesis versus co-culture on stiff substrates. As this example highlights, this technology better controls the in vitro microenvironment, allowing the user to develop a more thorough understanding of the combined effects of cell-cell and cell-matrix interactions. PMID:24061208

  15. Puerarin exhibits greater distribution and longer retention time in neurons than astrocytes in a co-cultured system

    Directory of Open Access Journals (Sweden)

    Shu-Yong Wei

    2015-01-01

    Full Text Available The phytoestrogen puerarin has been shown to protect neurons and astrocytes in the brain, and is therefore an attractive drug in the treatment of Alzheimer′s disease, Parkinson′s disease and cerebral ischemia. Whether puerarin exhibits the same biological processes in neurons and astrocytes in vitro has rarely been reported. In this study, cortical neurons and astrocytes of newborn Sprague-Dawley rats were separated, identified and co-cultured in a system based on Transwell membranes. The retention time and distribution of puerarin in each cell type was detected by fluorescence spectrophotometry and fluorescence microscope. The concentration of puerarin in both co-cultured and separately cultured neurons was greater than that of astrocytes. Puerarin concentration reached a maximum 20 minutes after it was added. At 60 minutes after its addition, a scant amount of drug was detected in astrocytes; however in both separately cultured and co-cultured neurons, the concentration of puerarin achieved a stable level of about 12.8 ng/mL. The results indicate that puerarin had a higher concentration and longer retention time in neurons than that observed in astrocytes.

  16. Histaminergic system in co-cultures of hippocampus and posterior hypothalamus: a morphological and electrophysiological study in the rat.

    Science.gov (United States)

    Diewald, L; Heimrich, B; Büsselberg, D; Watanabe, T; Haas, H L

    1997-11-01

    Neurons of the tuberomammillary nucleus in the posterior hypothalamus diffusely project to most parts of the central nervous system, where their main transmitter, histamine, modulates the excitability of the target neurons. The development of a histaminergic hypothalamo-hippocampal pathway and its function were studied in organotypic co-cultures. Immunocytochemistry for histidine decarboxylase, the specific synthesizing enzyme, stained clusters of neurons in the hypothalamic tuberomammillary area. Immunolabelled varicose processes innervated the co-cultured hippocampus and established a few synaptic contacts on dendrites. Cultured tuberomammillary neurons displayed their typical membrane properties and were spontaneously active. In hippocampal pyramidal cells of the CA3 region the long-lasting afterhyperpolarization was reduced by histamine or impromidine and increased by the H2 antagonist cimetidine, but not by the H1 antagonist mepyramine. The membrane potential was depolarized in presence of an H2 agonist and hyperpolarized by an H2 antagonist. In single hippocampal cultures histamine antagonists did not affect afterhyperpolarization and membrane potential. Histaminergic neurons retain their main morphological and physiological characteristics in slice cultures and establish a functional connection with co-cultured target cells.

  17. Towards the characterization of an in vitro triple co-culture intestine cell model for permeability studies.

    Science.gov (United States)

    Araújo, Francisca; Sarmento, Bruno

    2013-12-15

    Caco-2 based cell models have been the gold standard in vitro method to study intestinal drug permeability, despite the absence of many important features with major influence in the drug absorption mechanism. In the present work, a triple co-culture comprising Caco-2, HT29-MTX and Raji B cells was established to mimic in a closely way the human intestinal epithelium, presenting the main components in the process of drug absorption, namely the absorptive cells that resemble enterocytes, mucus producers cells and cells able to induce M-cell phenotype on Caco-2 cells. All the three cell lines maintained their function when cultured together with each other being, thus, an asset to new orally administrated drugs development. The seeding ratio of 90:10 between Caco-2 and HT29-MTX showed to be the best to achieve physiological proportions after cells maturation and differentiation in culture. The formation of M-cells phenotype from enterocytes was identified for the first time in a co-culture system comprising Caco-2 and HT29-MTX cells through immunocytochemical techniques. Thus, the triple co-culture model presented in the herein work is a good and reliable alternative to the in vitro methods already existents for the study of drugs permeability. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Development and characterization of a three-dimensional co-culture model of tumor T cell infiltration.

    Science.gov (United States)

    Alonso-Nocelo, M; Abuín, C; López-López, R; de la Fuente, M

    2016-04-14

    Tumor growth and metastasis entangle the alteration and recruitment of non-malignant cells to the primary tumor, among them immune cells, constituting the tumor microenvironment (TME). Communication between tumor cells and their stroma has been shown as a fundamental driving force of the tumoral process. A great deal of effort has been focused on depicting their specific interactions and crosstalk. However, most research has been carried out in 2D conventional cultures that alter cell morphology and intracellular signaling processes. Considering these premises, we have developed a 3D cell co-culture model to mimic T cell infiltration into the tumor mass and explore tumor-immune cells interactions in the TME. Expression of specific cell markers and assessment of cell proliferation were carried out to characterize the proposed 3D co-culture model. Additionally, the study and profiling of the secretome revealed a subset of particular cancer-related inflammation proteins prompted upon 3D cultivation of tumor cells in presence of lymphocytes, pointing out an intercellular communication. Altogether, these results suggest that our 3D cell co-culture model can be a useful tool to identify and study critical factors mediating the crosstalk between tumor and immune cells in the TME. Finally, the potential of this model as a drug-screening platform has been explored using docetaxel as a model antitumoral compound.

  19. Human ovarian neoplasm cell CD147 stimulates production and activation of matrix metalloproteinases in co-cultures with mouse fibroblasts

    Institute of Scientific and Technical Information of China (English)

    YANG Hong; ZOU Wei; XIN Xiao-yan

    2005-01-01

    Objective: To investigate the expression of CD147 on human ovarian neoplasm cell lines and its influence on production and activation of matrix metallproteinases(MMPs). Methods: The expression of CD147 on different human ovarian neoplasm cell lines was studied by western blotting. Co-culture was carried out to investigate the stimulative effect of the positive expression CD147 cell HO-8910 on the production of MMPs of fibroblast cell in vitro. Zymography and immune blotting were used to study the production and activity of positive MMPs, at the time, to explore the relation between CD147 and MMPs. Results: CD147 was positively presented in 2 ovarian neoplasm cell lines(HO-8910,3-AO), but in SKOV3, TC-1,NIN3T3 cell was negative. MMP-2 and MMP-9 were detected by HO-8910 cell line, mouse fibroblast cell and co-culture cells; but the expression in co-culture cell is obviously higher than individual cultures of each type alone.CD147 stimulated MMPs in dose-dependent manner. Conclusion: CD147 causes increased production and activation of MMP-2, MMP-9.CD147 is probably a indirect marker of some ovarian cancer cells with invasion and metastasis.

  20. Production of Fusaric Acid by Fusarium spp. in Pure Culture and in Solid Medium Co-Cultures

    Directory of Open Access Journals (Sweden)

    Nadine Bohni

    2016-03-01

    Full Text Available The ability of fungi isolated from nails of patients suffering from onychomycosis to induce de novo production of bioactive compounds in co-culture was examined. Comparison between the metabolite profiles produced by Sarocladium strictum, by Fusarium oxysporum, and by these two species in co-culture revealed de novo induction of fusaric acid based on HRMS. Structure confirmation of this toxin, using sensitive microflow NMR, required only three 9-cm Petri dishes of fungal culture. A targeted metabolomics study based on UHPLC-HRMS confirmed that the production of fusaric acid was strain-dependent. Furthermore, the detected toxin levels suggested that onychomycosis-associated fungal strains of the F. oxysporum and F. fujikuroi species complexes are much more frequently producing fusaric acid, and in higher amount, than strains of the F. solani species complex. Fusarium strains producing no significant amounts of this compound in pure culture, were shown to de novo produce that compound when grown in co-culture. The role of fusaric acid in fungal virulence and defense is discussed.

  1. Characterization of human lung cancer-associated fibroblasts in three-dimensional in vitro co-culture model

    Energy Technology Data Exchange (ETDEWEB)

    Horie, Masafumi [Department of Respiratory Medicine, Graduate School of Medicine, University of Tokyo (Japan); Saito, Akira, E-mail: asaitou-tky@umin.ac.jp [Department of Respiratory Medicine, Graduate School of Medicine, University of Tokyo (Japan); Mikami, Yu [Department of Respiratory Medicine, Graduate School of Medicine, University of Tokyo (Japan); Ohshima, Mitsuhiro [Department of Biochemistry, Ohu University School of Pharmaceutical Sciences (Japan); Morishita, Yasuyuki [Department of Molecular Pathology, Graduate School of Medicine, University of Tokyo (Japan); Nakajima, Jun [Department of Thoracic Surgery, Graduate School of Medicine, University of Tokyo (Japan); Kohyama, Tadashi; Nagase, Takahide [Department of Respiratory Medicine, Graduate School of Medicine, University of Tokyo (Japan)

    2012-06-22

    Highlights: Black-Right-Pointing-Pointer We established three patient-paired sets of CAFs and NFs. Black-Right-Pointing-Pointer CAFs and NFs were analyzed using three-dimensional co-culture experiments. Black-Right-Pointing-Pointer CAFs clearly enhanced collagen gel contraction. Black-Right-Pointing-Pointer CAFs showed higher {alpha}-SMA expression than NFs. Black-Right-Pointing-Pointer CAFs were implicated in invasion and differentiation of lung cancer cells. -- Abstract: Lung cancer is the most common cause of cancer-related death worldwide. Stromal cancer-associated fibroblasts (CAFs) play crucial roles in carcinogenesis, proliferation, invasion, and metastasis of non-small cell lung carcinoma, and targeting of CAFs could be a novel strategy for cancer treatment. However, the characteristics of human CAFs still remain to be better defined. In this study, we established patient-matched CAFs and normal fibroblasts (NFs), from tumoral and non-tumoral portions of resected lung tissue from lung cancer patients. CAFs showed higher {alpha}-smooth muscle actin ({alpha}-SMA) expression than NFs, and CAFs clearly enhanced collagen gel contraction. Furthermore, we employed three-dimensional co-culture assay with A549 lung cancer cells, where CAFs were more potent in inducing collagen gel contraction. Hematoxylin and eosin staining of co-cultured collagen gel revealed that CAFs had the potential to increase invasion of A549 cells compared to NFs. These observations provide evidence that lung CAFs have the tumor-promoting capacity distinct from NFs.

  2. Comparative evaluation of the hydrogen production by mixed consortium, synthetic co-culture and pure culture using distillery effluent.

    Science.gov (United States)

    Mishra, Preeti; Roy, Shantonu; Das, Debabrata

    2015-12-01

    Wastewater comprises of various carbon sources. So, the use of microbial consortium may improve the hydrogen production and organic reduction. The present study deals with biohydrogen production by acidogenic mixed consortia (AMC), synthetic co-culture (Klebsiella pneumoniae IIT-BT 08 and Citrobacter freundii IIT-BT L139) and pure culture using distillery effluent (DE). Higher hydrogen yield was observed in case of AMC (9.17 mol/kg CODreduced) as compared to the synthetic co-culture and pure culture. PCR-DGGE analysis indicated that the consortium was predominated by species closely affiliated to Clostridium sp. The average hydrogen production rate was 267 mL/Lh. The maximum hydrogen production rate (Rm), hydrogen production potential (P) and lag time (λ) by AMC using DE were 507.2 mL/Lh, 3729 m/L and 2.04 h, respectively. Maximum gaseous energy recovery by AMC was found to be higher by 21.9% and 45.4% than that of using co-culture and pure culture respectively.

  3. Enterococcus faecium RZS C5, an interesting bacteriocin producer to be used as a co-culture in food fermentation.

    Science.gov (United States)

    Leroy, F; Foulquié Moreno, M R; De Vuyst, L

    2003-12-01

    Enterocins, bacteriocins produced by enterococci, are gaining interest because of their industrial potential. Due to its bacteriocin production, Enterococcus faecium RZS C5, a natural cheese isolate, has a strong activity towards Listeria monocytogenes. For this reason, the strain may be applicable as a bacteriocin-producing co-culture in food fermentation in order to reduce the risk on Listeria outgrowth. The strain displays remarkable bacteriocin production kinetics. Whereas most lactic acid bacteria produce bacteriocin in a growth-associated way until the beginning of the stationary phase, bacteriocin production by E. faecium RZS C5 in MRS broth at controlled pH values below 7.5 is characterised by a boost of bacteriocin activity levels in the very early growth phase. In addition, bacteriocin production kinetics are closely linked to the environmental and cultural conditions. However, no straightforward statement about the effect of environmental stress on bacteriocin production can be made since the effect is dependent on the type of stress applied. Kinetic experiments in milk and on pilot scale, applying Cheddar cheese-making conditions, have indicated that the strain may be effective as a bacteriocin-producing co-culture. Further research is needed to evaluate the use of E. faecium RZS C5 as a co-culture for the production of fermented sausage.

  4. Effects of intermittent hydrostatic pressure magnitude on the chondrogenesis of MSCs without biochemical agents under 3D co-culture.

    Science.gov (United States)

    Jeong, Jae Young; Park, So Hee; Shin, Ji Won; Kang, Yun Gyeong; Han, Ki-Ho; Shin, Jung-Woog

    2012-11-01

    Without using biochemical agents, in this study, we sought to investigate the potential of controlling the differentiation of mesenchymal stem cells (MSCs) into a specific cell type through the use of 3D co-culturing and mechanical stimuli. MSCs and primary cultured chondrocytes were separately encapsulated into alginate beads, and the two types of beads were separated by a membrane. For the investigation a computer-controllable bioreactor was designed and used to engage intermittent hydrostatic pressure (IHP). Five different magnitudes (0.20, 0.10, 0.05, 0.02 MPa and no stimulation) of IHP were applied. The stimulation pattern was the same for all groups: 2 h/day for 7 days starting at 24 h after seeding; 2 and 15 min cycles of stimulating and resting, respectively. Biochemical (DNA and GAG contents), histological (Alcian blue), and RT-PCR (Col II, SOX9, AGC) analyses were performed on days 1, 5, 10, and 20. The results from these analyses showed that stimulation with higher magnitudes of IHP (≥0.10 MPa) were more effective on the proliferation and differentiation of co-cultured MSCs. Together, these data demonstrate the potential of using mechanical stimulation and co-culturing for the proliferation and differentiation of MSCs, even without biochemical agents.

  5. Simultaneous and quantitative monitoring of co-cultured Pseudomonas aeruginosa and Staphylococcus aureus with antibiotics on a diffusometric platform

    Science.gov (United States)

    Chung, Chih-Yao; Wang, Jhih-Cheng; Chuang, Han-Sheng

    2017-04-01

    Successful treatments against bacterial infections depend on antimicrobial susceptibility testing (AST). However, conventional AST requires more than 24 h to obtain an outcome, thereby contributing to high patient mortality. An antibiotic therapy based on experiences is therefore necessary for saving lives and escalating the emergence of multidrug-resistant pathogens. Accordingly, a fast and effective drug screen is necessary for the appropriate administration of antibiotics. The mixed pathogenic nature of infectious diseases emphasizes the need to develop an assay system for polymicrobial infections. On this basis, we present a novel technique for simultaneous and quantitative monitoring of co-cultured microorganisms by coupling optical diffusometry with bead-based immunoassays. This simple integration simultaneously achieves a rapid AST analysis for two pathogens. Triple color particles were simultaneously recorded and subsequently analyzed by functionalizing different fluorescent color particles with dissimilar pathogen-specific antibodies. Results suggested that the effect of the antibiotic, gentamicin, on co-cultured Pseudomonas aeruginosa and Staphylococcus aureus was effectively distinguished by the proposed technique. This study revealed a multiplexed and time-saving (within 2 h) platform with a small sample volume (~0.5 μL) and a low initial bacterial count (50 CFU per droplet, ~105 CFU/mL) for continuously monitoring the growth of co-cultured microorganisms. This technique provides insights into timely therapies against polymicrobial diseases in the near future.

  6. Production of Fusaric Acid by Fusarium spp. in Pure Culture and in Solid Medium Co-Cultures.

    Science.gov (United States)

    Bohni, Nadine; Hofstetter, Valérie; Gindro, Katia; Buyck, Bart; Schumpp, Olivier; Bertrand, Samuel; Monod, Michel; Wolfender, Jean-Luc

    2016-03-18

    The ability of fungi isolated from nails of patients suffering from onychomycosis to induce de novo production of bioactive compounds in co-culture was examined. Comparison between the metabolite profiles produced by Sarocladium strictum, by Fusarium oxysporum, and by these two species in co-culture revealed de novo induction of fusaric acid based on HRMS. Structure confirmation of this toxin, using sensitive microflow NMR, required only three 9-cm Petri dishes of fungal culture. A targeted metabolomics study based on UHPLC-HRMS confirmed that the production of fusaric acid was strain-dependent. Furthermore, the detected toxin levels suggested that onychomycosis-associated fungal strains of the F. oxysporum and F. fujikuroi species complexes are much more frequently producing fusaric acid, and in higher amount, than strains of the F. solani species complex. Fusarium strains producing no significant amounts of this compound in pure culture, were shown to de novo produce that compound when grown in co-culture. The role of fusaric acid in fungal virulence and defense is discussed.

  7. In vitro and in vivo co-culture of chondrocytes and bone marrow stem cells in photocrosslinked PCL-PEG-PCL hydrogels enhances cartilage formation.

    Science.gov (United States)

    Ko, Chao-Yin; Ku, Kuan-Lin; Yang, Shu-Rui; Lin, Tsai-Yu; Peng, Sydney; Peng, Yu-Shiang; Cheng, Ming-Huei; Chu, I-Ming

    2016-10-01

    Chondrocytes (CH) and bone marrow stem cells (BMSCs) are sources that can be used in cartilage tissue engineering. Co-culture of CHs and BMSCs is a promising strategy for promoting chondrogenic differentiation. In this study, articular CHs and BMSCs were encapsulated in PCL-PEG-PCL photocrosslinked hydrogels for 4 weeks. Various ratios of CH:BMSC co-cultures were investigated to identify the optimal ratio for cartilage formation. The results thus obtained revealed that co-culturing CHs and BMSCs in hydrogels provides an appropriate in vitro microenvironment for chondrogenic differentiation and cartilage matrix production. Co-culture with a 1:4 CH:BMSC ratio significantly increased the synthesis of GAGs and collagen. In vivo cartilage regeneration was evaluated using a co-culture system in rabbit models. The co-culture system exhibited a hyaline chondrocyte phenotype with excellent regeneration, resembling the morphology of native cartilage. This finding suggests that the co-culture of these two cell types promotes cartilage regeneration and that the system, including the hydrogel scaffold, has potential in cartilage tissue engineering. Copyright © 2013 John Wiley & Sons, Ltd.

  8. Clarification of interactions among microorganisms and development of co-culture system; Biseibutsukan sokosayo no kaiseki to kongo baiyo shisutemu no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    Taniguchi, Masayuki [Niigata University, Niigata (Japan). Dept. of Materials Science and Technology

    1999-03-10

    Co-culture systems containing two microorganisms for production of useful substances are described. The co-culture of Bifidobacterium longum and Propionibacterium freudenreichii, where lactic acid produced once from lactose by B. longum is converted to acetic and propionic acids by P. freudenreichii, was carried out. Thought the sequential conversion of lactose using the two microorganisms, the culture supernatant containing a mixture of acetic and propionic acids without lactic acid is produced. The antimicrobial activity of the mixture is higher than that obtained in the cultivation of B. longum alone. We developed a novel co-culture system composed of two fermentors and two micro filtration modules for efficient ethanol production from a mixture of glucose and xylose by co-culture of Pichia stipitis and Saccharomyces cerevisiae. The proposed co-culture system allowed regulation of the dissolved oxygen concentration at a level suitable for an individual yeast in each fermentor, as well as the successful exchange of culture medium between two fermentors. When P. stipitis and S. cerevisiae are cultivated individually under different oxygen supply conditions in the new co-culture system, the yield and productivity of ethanol from a glucose and xylose mixture are higher than in single culture of P. stipitis alone. By clarifying the interactions among microorganisms, new bioprocesses in which similar performance to co-culture systems is expressed even using a single microorganism, are expected to be developed for improvement of biochemical reaction systems. (author)

  9. 气单胞菌和嗜水气单胞菌双重PCR检测方法的建立%Development of a duplex PCR for detection of Aeromonas spp.and Aeromonas hydrophila

    Institute of Scientific and Technical Information of China (English)

    潘晓艺; 沈锦玉; 郝贵杰; 姚嘉贇; 徐洋; 尹文林; 孙逢明; 吴颖蕾

    2011-01-01

    Aeromonas is one of the major pathogens which frequently cause the bacterial diseases in freshwater fish terrapin and crustaceans. Aeromonas hydrophila caused economic losses widely. In order to detect Aeromonas spp. And Aeromonas hydrophila, two pairs of specific primers were designed according to the glycerophospholipid-cholesterol acyltransferase (GCAT) gene of Aeromonas spp. And the 16S rRNA gene of Aeromonas hydrophila. So a duplex PCR assay was developed to amplify the 16S rRNA gene and the GCAT gene. The reaction conditions of the duplex PCR were optimized and PCR products were sequenced. Meanwhile, the specificity of duplex PCR was studied. Five Aeromonas hydrophila strains,seven Aeromonas strains and the other five strains of non Aeromonas were tested by duplex PCR. The results showed that the GCAT gene and the 16S rRNA gene could be amplified from Aeromonas hydrophila strains,but only the GCAT gene from the other Aeromonas species and neither from the other five strains. It could be concluded that the duplex PCR was specific and could be used in quick diagnose Aeromonas hydrophila and Aeromonas spp.%针对GenBank中登录的气单胞菌属(Aeromonas)的毒力基因甘油磷脂胆固醇酰基转移酶基因(GCA T)和嗜水气单胞菌(Aeromonash ydrophila)的16S rRNA基因的保守区设计2对特异性引物.通过进行双重PCR反应体系优化,PCR产物的测序鉴定和特异性试验,建立了一种能同时检测气单胞菌(Aeromonas spp.)和嗜水气单胞菌的双重PCR检测方法.用此方法对5株嗜水气单胞菌、7株其他不同种的气单胞菌和5株非气单胞菌属菌株进行双重PCR检测.结果显示,气单胞菌和嗜水气单胞菌在GCAT基因的扩增区都能得到有效扩增,其中嗜水气单胞菌能同时在16S rRNA基因扩增区得到有效扩增,而非气单胞菌属的菌株,在此两扩增区都为阴性,表明此检测方法可靠且可行.

  10. [Influence of predominant aerobic bacteria isolated from different healthy animals on daidzein biotransforming capacity by co-culture with different daidzein biotransforming bacteria].

    Science.gov (United States)

    Luo, Jinglong; Wang, Xiuling; Fan, Jinru; Wang, Shiying; Li, Jia

    2011-08-01

    To investigate the influence of isolated predominant aerobic bacteria on daidzein biotransformation capacity by co-culture with daidzein biotransforming bacteria. Predominant aerobic bacteria were isolated from diluted feces solutions of different healthy animals, including ICR mice, Luhua chicken, Landrace pigs and Rex rabbits. Daidzein biotransforming bacteria were anaerobically co-cultured with the isolated predominant aerobic bacteria and the cultural broth was extracted and detected by high performance liquid chromatography (HPLC). Twenty two predominant aerobic bacteria were isolated from the four different healthy animals mentioned above. Based on the analyses of 16S rRNA gene sequences, morphology study and relative biophysico-biochemical characteristics, all 22 isolates belong to the 5 genera, i. e. Escherichia (10) , Proteus (5) , Enterococcus (4), Bacillus (2) and Pseudomonas (1). Co-culture between predominant aerobic bacteria and daidzein biotransforming bacteria was carried out under anaerobic conditions. The results showed that the biotransformation capacity was totally lost when different daidzein biotransforming bacterium was co-cultured with either Bacillus cereus ( R1 ) or Pseudomonas aerginosa (R5) and continuously inoculated for 2 or 3 passages. However, no obvious influence was observed when daidzein biotransforming bacteria were co-cultured with all the other isolated predominant aerobic bacteria except R1 and R5. In addition, when strain R1 and R5 was co-cultured with the intestinal microflora of the ICR mice anaerobically and continuously inoculated for 5 passages, about 90% of the co-cultures totally lost the activity to convert daidzein to equol effectively. Different predominant aerobic bacteria showed different influence on daidzein biotransformation capacity after being co-cultured with different daidzein biotransforming bacteria. Among all the isolated predominant aerobic bacteria used for co-culture, both Bacillus cereus ( R1) and

  11. Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods

    Directory of Open Access Journals (Sweden)

    Akiko Edagawa

    2015-10-01

    Full Text Available We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR, and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%. Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%. In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8% compared with real-time qPCR alone (46/68, 67.6%. Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1% compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%. Legionella was not detected in the remaining six samples (6/68, 8.8%, irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

  12. Investigation of Legionella Contamination in Bath Water Samples by Culture, Amoebic Co-Culture, and Real-Time Quantitative PCR Methods.

    Science.gov (United States)

    Edagawa, Akiko; Kimura, Akio; Kawabuchi-Kurata, Takako; Adachi, Shinichi; Furuhata, Katsunori; Miyamoto, Hiroshi

    2015-10-19

    We investigated Legionella contamination in bath water samples, collected from 68 bathing facilities in Japan, by culture, culture with amoebic co-culture, real-time quantitative PCR (qPCR), and real-time qPCR with amoebic co-culture. Using the conventional culture method, Legionella pneumophila was detected in 11 samples (11/68, 16.2%). Contrary to our expectation, the culture method with the amoebic co-culture technique did not increase the detection rate of Legionella (4/68, 5.9%). In contrast, a combination of the amoebic co-culture technique followed by qPCR successfully increased the detection rate (57/68, 83.8%) compared with real-time qPCR alone (46/68, 67.6%). Using real-time qPCR after culture with amoebic co-culture, more than 10-fold higher bacterial numbers were observed in 30 samples (30/68, 44.1%) compared with the same samples without co-culture. On the other hand, higher bacterial numbers were not observed after propagation by amoebae in 32 samples (32/68, 47.1%). Legionella was not detected in the remaining six samples (6/68, 8.8%), irrespective of the method. These results suggest that application of the amoebic co-culture technique prior to real-time qPCR may be useful for the sensitive detection of Legionella from bath water samples. Furthermore, a combination of amoebic co-culture and real-time qPCR might be useful to detect viable and virulent Legionella because their ability to invade and multiply within free-living amoebae is considered to correlate with their pathogenicity for humans. This is the first report evaluating the efficacy of the amoebic co-culture technique for detecting Legionella in bath water samples.

  13. Stable coexistence of two Caldicellulosiruptor species in a de novo constructed hydrogen-producing co-culture

    Directory of Open Access Journals (Sweden)

    Rådström Peter

    2010-12-01

    Full Text Available Abstract Background Mixed culture enrichments have been used frequently for biohydrogen production from different feedstock. In spite of the several advantages offered by those cultures, they suffer poor H2 yield. Constructing defined co-cultures of known H2 producers may offer a better performance than mixed-population enrichments, while overcoming some of the limitations of pure cultures based on synergies among the microorganisms involved. Results The extreme thermophiles Caldicellulosiruptor saccharolyticus DSM 8903 and C. kristjanssonii DSM 12137 were combined in a co-culture for H2 production from glucose and xylose in a continuous-flow stirred tank reactor. The co-culture exhibited a remarkable stability over a period of 70 days under carbon-sufficient conditions, with both strains coexisting in the system at steady states of different dilution rates, as revealed by species-specific quantitative PCR assays. The two strains retained their ability to stably coexist in the reactor even when glucose was used as the sole growth-limiting substrate. Furthermore, H2 yields on glucose exceeded those of either organism alone under the same conditions, alluding to a synergistic effect of the two strains on H2 production. A maximum H2 yield of 3.7 mol (mol glucose-1 was obtained by the co-culture at a dilution rate of 0.06 h-1; a higher yield than that reported for any mixed culture to date. A reproducible pattern of population dynamics was observed in the co-culture under both carbon and non-carbon limited conditions, with C. kristjanssonii outgrowing C. saccharolyticus during the batch start-up phase and prevailing at higher dilution rates. A basic continuous culture model assuming the ability of C. saccharolyticus to enhance the growth of C. kristjanssonii could mimic the pattern of population dynamics observed experimentally and provide clues to the nature of interaction between the two strains. As a proof, the cell-free growth supernatant of C

  14. Co-culture of C2C12 and 3T3-L1 preadipocyte cells alters the gene expression of calpains, caspases and heat shock proteins.

    Science.gov (United States)

    Pandurangan, Muthuraman; Jeong, Dawoon; Amna, Touseef; Van Ba, Hoa; Hwang, Inho

    2012-10-01

    The present study was carried out to understand the co-culture effect of C2C12 and 3T3-L1 preadipocyte cells on calpain, caspase, and heat shock protein (Hsp) systems. Calpains, caspases, and heat shock proteins play critical roles in the growth and development of mammalian cells. Cells were co-cultured using transwell inserts with a 0.4-μm porous membrane to separate C2C12 and 3T3-L1 preadipocyte cells. Each cell type was grown independently on the transwell plates. Following cell differentiation, inserts containing 3T3-L1 cells were transferred to C2C12 plates and inserts containing C2C12 transferred to 3T3-L1 plates. Following co-culture for 24 and 48 h, the cells in the lower well were harvested for analysis. Calpains include μ-calpain, m-calpain, and their specific inhibitor calpastatin. The expression pattern of μ-calpain did not change in the co-cultured C2C12 and 3T3-L1 cells, whereas m-capain mRNA expression significantly reduced in the 48-h co-cultured 3T3-L1 cells. Calpastatin mRNA expression significantly increased in the 48-h co-cultured C2C12 cells. Caspase-7 mRNA expression did not change in the 24- and 48-h co-cultured C2C12 and 3T3-L1 cells. Caspase-3 mRNA expression significantly reduced in the 24- and 48-h co-cultured 3T3-L1 cells; caspase-9 mRNA had a significant reduction only at 48 h, whereas caspase-9 mRNA expression significantly increased in the 48-h co-cultured C2C12 cells. Hsp27 and Hsp90 mRNA expressions are significantly reduced in the 24- and 48-h co-cultured C2C12 and 3T3-L1 cells, whereas Hsp70 mRNA expression significantly increased in the 48-h co-cultured 3T3-L1 cells. The co-culture reflects three-dimensional views of C2C12 and 3T3-L1 cell types as in vivo, which is quite distinct from the one-dimensional monocultured C2C12 and 3T3-L1 cells.

  15. Effects of Substrate and Co-Culture on Neural Progenitor Cell Differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Jones, Erin Boote [Iowa State Univ., Ames, IA (United States)

    2008-01-01

    In recent years the study of stem and progenitor cells has moved to the forefront of research. Since the isolation of human hematopoietic stem cells in 1988 and the subsequent discovery of a self renewing population of multipotent cells in many tissues, many researchers have envisioned a better understanding of development and potential clinical usage in intractable diseases. Both these goals, however, depend on a solid understanding of the intracellular and extracellular forces that cause stem cells to differentiate to a specific cell fate. Many diseases of large scale cell loss have been suggested as candidates for stem cell based treatments. It is proposed that replacing the function of the damaged or defective cells by specific differentiation of stem or progenitor cells could treat the disease. Before cells can be directed to specific lineages, the mechanisms of differentiation must be better understood. Differentiation in vivo is an intensively complex system that is difficult to study. The goal of this research is to develop further understanding of the effects of soluble and extracellular matrix (ECM) cues on the differentiation of neural progenitor cells with the use of a simplified in vitro culture system. Specific research objectives are to study the differentiation of neural progenitor cells in response to astrocyte conditioned medium and protein substrate composition and concentration. In an effort to reveal the mechanism of the conditioned medium interaction, a test for the presence of a feedback loop between progenitor cells and astrocytes is presented along with an examination of conditioned medium storage temperature, which can reveal enzymatic dependencies. An examination of protein substrate composition and concentration will help to reveal the role of any ECM interactions on differentiation. This thesis is organized into a literature review covering recent advances in use of external modulators of differentiation such as surface coatings, co-culture

  16. Probiotic modulation of dendritic cells co-cultured with intestinal epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Ji Yeun Kim; Myeong Soo Park; Geun Eog Ji

    2012-01-01

    AIM:TO investigate cytokine production and cell surface phenotypes of dendritic cells (DC) in the presence of epithelial cells stimulated by probiotics.METHODS:Mouse DC were cultured alone or together with mouse epithelial cell monolayers in normal or inverted systems and were stimulated with heat-killed probiotic bacteria,Bifidobacteriumlactis AD011 (BL),Bifidobacterium bifidum BGN4 (BB),Lactobacillus casei IBS041 (LC),and Lactobacillus acidophilus AD031 (LA),for 12 h.Cytokine levels in the culture supernatants were determined by enzyme-linked immunosorbent assay and phenotypic analysis of DC was investigated by flow cytometry.RESULTS:BB and LC in single-cultured DC increased the expression of I-Ad,CD86 and CD40 (I-Ad,18.51 vs 30.88,46.11; CD86,62.74 vs 92.7,104.12; CD40,0.67vs 6.39,3.37,P < 0.05).All of the experimental probiotics increased the production of inflammatory cytokines,interleukin (IL)-6 and tumor necrosis factor (TNF)-α.However,in the normal co-culture systems,LC and LA decreased the expression of I-Ad (39.46 vs 30.32,33.26,P < 0.05),and none of the experimental probiotics increased the levels of IL-6 or TNF-α.In the inverted coculture systems,LC decreased the expression of CD40 (1.36 vs-2.27,P < 0.05),and all of the experimental probiotics decreased the levels of IL-6.In addition,BL increased the production of IL-10 (103.8 vs 166.0,P< 0.05) and LC and LA increased transforming growth factor-3 secretion (235.9 vs 618.9,607.6,P < 0.05).CONCLUSION:These results suggest that specific probiotic strains exert differential immune modulation mediated by the interaction of dendritic cells and epithelial cells in the homeostasis of gastrointestinal tract.

  17. Reação em cadeia da polimerase para detecção de Clostridium chauvoei em tecidos de Cavia porcellus PCR detection of Clostridium chauvoei in tissues of Cavia porcellus

    Directory of Open Access Journals (Sweden)

    Ronnie Antunes de Assis

    2005-12-01

    Full Text Available O objetivo deste trabalho foi padronizar uma técnica de reação em cadeia da polimerase (PCR, para detecção de Clostridium chauvoei, em tecidos fixados em formol e incluídos em parafina de cobaias (Cavia porcellus experimentalmente infectadas com esse microrganismo. Os animais foram sacrificados, e amostras do músculo da área de inoculação (MAI, fígado, miocárdio e baço foram disponibilizadas para a técnica de PCR. O clostrídio foi detectado em todas as secções do MAI, fígado e miocárdio, mas não foi observado em secções do baço. Reações cruzadas não foram observadas a partir de secções do MAI dos animais com inoculação de outras espécies de clostrídios, bem como nenhuma amplificação foi observada a partir de secções do MAI dos animais controle. Esses resultados mostram que a técnica de PCR desenvolvida neste estudo, pode ser usada para detecção de Clostridium chauvoei em tecidos fixados em formol e incluídos em parafina.The objective of this work was the standardization of a polymerase chain reaction (PCR for detection of Clostridium chauvoei in formalin-fixed, paraffin-embedded tissues of guinea-pigs (Cavia porcellus infected experimentally with this microorganism. The animals were sacrificed, and samples of muscle from inoculation area (MIA, liver, myocardium and spleen were available for PCR technique. Clostridium chauvoei was detected in all sections of the MIA, liver and myocardium, and no product was observed in sections of the spleen. Cross-reactions were not observed in sections of MIA of the animals inoculated with other clostridia, as well as no amplification was observed in sections of MIA of control animals. These results show that the PCR technique developed in this study may be useful for detection of C. chauvoei in formalin-fixed, paraffin-embedded tissues.

  18. 基于流动性调整CAViaR模型的风险度量方法%A New Method of Risk Measurement Based on Liquidity Adjusted CAViaR Models

    Institute of Scientific and Technical Information of China (English)

    闫昌荣

    2012-01-01

    The measurement and management of the liquidity risk, which is one of the major risks faced by investors, has always been one of the most difficult problems in both academia and practice. In this paper, we propose a new risk measurement method, called liquidity adjusted CAViaR models, to help investors manage future risks better, especially liquidity risk. This method can directly reflect the impacts of liquidity changes on the future risks, and the future liquidity adjusted VaR can be calculated simultanuously. Empirical studies suggest that this model can characterize the behavior of dynamic changes of liquidity risks in Chinese stock market, and conclude that a substantial decline of stock liquidity may lead to a significant increase of future risks. The positive liquidity may have a more significant effect than that of negative liquidity, thus it is more worthy of attentions from investors.%本文提出了流动性风险度量的一个新的方法,流动性调整的CAViaR模型。该模型能够直接反映资产流动性的变动对未来风险的影响,并在此基础上计算资产未来经过流动性调整的风险VaR,从而使投资者能够更好地管理风险,尤其是流动性风险。实证研究表明,该模型能够较好地刻画中国股市流动性风险的动态变化特征;并且发现股票流动性的大幅下降通常导致未来风险明显加大,且正向流动性下降所带来的风险往往较负向流动性要更大,因此更值得投资者关注。

  19. Endoparasitos em cobaias (Cavia porcellus (Mammalia, Rodentia, Caviidae provenientes de biotérios de criação e experimentação do município do Rio de Janeiro, Brasil Endoparasites in guinea pigs (Cavia porcellus (Mammalia, Rodentia, Caviidae from breeding and experimentation animal housing of the municipality of Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Luciana Casartelli Alves

    2007-10-01

    Full Text Available Foi realizado um levantamento sobre a prevalência e intensidade de infecção de endoparasitos em cobaias convencionais de linhagem Short Hair provenientes de biotérios de criação (A e experimentação (B do município do Rio de Janeiro, Brasil, avaliando-se a eficácia das medidas de prevenção entre eles. Para a realização do estudo, utilizou-se exame direto de mucosa e do conteúdo intestinal, a técnica de tricromo de WHEATLEY e exames coproparasitológicos pelas técnicas de Ritchie e Kinyoun. Os parasitos encontrados através da técnica de exame direto da mucosa e do conteúdo intestinal e de tricromo de WHEATLEY com as respectivas prevalências foram: Balantidium sp. (78%, Cyathodinium sp. (68%, Eimeria caviae (38%, Paraspidodera uncinata (34% e Giardia muris (24%. Nos exames de fezes realizados pelas técnicas de Ritchie e Kinyoun, foram encontrados os seguintes parasitos com as respectivas prevalências no biotério A: E. caviae (74%, Balantidium sp. (68%, Cyathodinium sp. (68% e Cryptosporidium sp. (5%. No biotério B, observou-se: E. caviae (58%, Balantidium sp. (42%, Cyathodinium sp. (25% e G. muris (8%. A alta prevalência de endoparasitos nos biotérios sugere a necessidade de se rever a eficácia das barreiras sanitárias adotadas.This paper discusses the prevalence and intensity of infection of endoparasites in conventionally maintained Short Hair guinea pigs colonies from a breeding (A and an experimental (B facilities in Rio de Janeiro, Brazil. It also evaluates the efficacy of the methods of prevention adopted by both facilities. The search of parasites was performed by direct examination of intestinal mucosa and its contents, WHEATLEY's trichrome method and coproparasitological examinations by Ritchie's and Kinyoun's techniques. The prevalences of endoparasites found throught direct examination of intestinal mucosa and its contents were: Balantidium sp. (78%, Cyathodinium sp. (68%, Eimeria caviae (38%, Paraspidodera

  20. Aeromonas sp.F3胶原酶性质的研究%Characteristies of collagenase from Aeromonas sp.F3

    Institute of Scientific and Technical Information of China (English)

    白杨; 钱斯日古楞; 王红英; 马蕾

    2011-01-01

    Aeromonas sp. F3 was screened from marine mud for hydrolyzing collagen. The characteristics of collagenase from Aeromonas sp. F3 were investigated via single-factor test. Thermal treatment at 50 ℃ for 40 min resulted in obvious deactivation. The optimum reaction temperature and pH was 40 ℃ and 8. 6. Collagenase activity was activated by Ca2+ at 0. 5 mmol/L. The molecular weight of the hydrolyzate was under 30 ku.%海洋污泥中筛选得到的Aeromonas sp.F3所产的胞外酶对胶原蛋白有水解作用.以海洋微生物Aeromonas sp.F3为酶源,采用单因素试验法对其所产的胶原酶性质进行了研究,包括该胶原酶的酶解条件及热稳定性,并对该胶原酶水解鱼皮的效果进行了分析.结果表明,微生物Aeromonas sp.F3源胶原酶在50℃热处理40 min之后明显失活,该胶原酶的最适反应温度为40℃,最适pH为8.6,金属离子Ca2在0.5 mmol/L时对酶有激活作用.该酶在其最适条件下,对鱼皮胶原有显著水解效能,其水解产物的分子质量在30 ku以下.

  1. Co-culture of oligodendrocytes and neurons can be used to assess drugs for axon regeneration in the central nervous system.

    Science.gov (United States)

    Gang, Lin; Yao, Yu-Chen; Liu, Ying-Fu; Li, Yi-Peng; Yang, Kai; Lu, Lei; Cheng, Yuan-Chi; Chen, Xu-Yi; Tu, Yue

    2015-10-01

    We present a novel in vitro model in which to investigate the efficacy of experimental drugs for the promotion of axon regeneration in the central nervous system. We co-cultured rat hippocampal neurons and cerebral cortical oligodendrocytes, and tested the co-culture system using a Nogo-66 receptor antagonist peptide (NEP1-40), which promotes axonal growth. Primary cultured oligodendrocytes suppressed axonal growth in the rat hippocampus, but NEP1-40 stimulated axonal growth in the co-culture system. Our results confirm the validity of the neuron-oligodendrocyte co-culture system as an assay for the evaluation of drugs for axon regeneration in the central nervous system.

  2. Response surface optimisation for acetone-butanol-ethanol production from cassava starch by co-culture of Clostridium butylicum and Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Benjamas Cheirsilp

    2011-11-01

    Full Text Available Acetone-butanol-ethanol (ABE production from cassava starch was enhanced by a syntrophic co-culture of Clostridium butylicum TISTR 1032 and high amylase producing Bacillus subtilis WD 161 without anaerobic pretreatment. The production of amylase and ABE using this co-culture were respectively 16 and 6 times higher than those using the pure culture of C. butylicum TISTR 1032. The effect of the medium components on the performance of the co-culture was investigated using response surface methodology (RSM. Among the investigated components, cassava starch and ammonium nitrate contributed a significant effect on the production of amylase and ABE, while yeast extract had less effect. Based on the optimum strategy using RSM, the ABE production by the co-culture was improved 2.2-fold compared with that obtained from the initial condition and with a minimum requirement of nitrogen source.

  3. Enhancing Butanol Production under the Stress Environments of Co-Culturing Clostridium acetobutylicum/Saccharomyces cerevisiae Integrated with Exogenous Butyrate Addition

    National Research Council Canada - National Science Library

    Luo, Hongzhen; Ge, Laibing; Zhang, Jingshu; Zhao, Yanli; Ding, Jian; Li, Zhigang; He, Zhenni; Chen, Rui; Shi, Zhongping

    2015-01-01

    In this study, an efficient acetone-butanol-ethanol (ABE) fermentation strategy integrating Clostridium acetobutylicum/Saccharomyces cerevisiae co-culturing system with exogenous butyrate addition, was proposed and experimentally conducted...

  4. Co-culture of oligodendrocytes and neurons can be used to assess drugs for axon regeneration in the central nervous system

    Directory of Open Access Journals (Sweden)

    Lin Gang

    2015-01-01

    Full Text Available We present a novel in vitro model in which to investigate the efficacy of experimental drugs for the promotion of axon regeneration in the central nervous system. We co-cultured rat hippocampal neurons and cerebral cortical oligodendrocytes, and tested the co-culture system using a Nogo-66 receptor antagonist peptide (NEP1-40, which promotes axonal growth. Primary cultured oligodendrocytes suppressed axonal growth in the rat hippocampus, but NEP1-40 stimulated axonal growth in the co-culture system. Our results confirm the validity of the neuron-oligodendrocyte co-culture system as an assay for the evaluation of drugs for axon regeneration in the central nervous system.

  5. Identification and molecular characterization of the homogentisate pathway responsible for pyomelanin production, the major melanin constituents in Aeromonas media WS.

    Science.gov (United States)

    Wang, He; Qiao, Yunqian; Chai, Baozhong; Qiu, Chenxi; Chen, Xiangdong

    2015-01-01

    The pigmentation of many Aeromonas species has been thought to be due to the production of a L-DOPA (L-3,4-dihydroxyphenylalanine) based melanin. However, in this study we found that although L-DOPA synthesis occurs in the high-melanin-yielding Aeromonas media strain WS, it plays a minor, if any, role in pigmentation. Instead, the pigmentation of A. media strain WS is due to the production of pyomelanin through HGA (homogentisate). Gene products of phhA (encodes phenylalanine hydroxylase), tyrB and aspC (both encode aromatic amino acid aminotransferase), and hppD (encodes 4-hydroxyphenylpyruvate dioxygenase) constitute a linear pathway of converting phenylalanine to HGA and disruption of any one of these genes impairs or blocks pigmentation of A. media strain WS. This HGA biosynthesis pathway is widely distributed in Aeromonas, but HGA is only detectable in the cultures of pigmented Aeromonas species. Heterologous expression of HppD from both pigmented and non-pigmented Aeromonas species in E. coli leads to the production of pyomelanin and thus pigmentation, suggesting that most Aeromonas species have the critical enzymes to produce pyomelanin through HGA. Taken together, we have identified a widely conserved biosynthesis pathway of HGA based pyomelanin in Aeromonas that may be responsible for pigmentation of many Aeromonas species.

  6. Identification and molecular characterization of the homogentisate pathway responsible for pyomelanin production, the major melanin constituents in Aeromonas media WS.

    Directory of Open Access Journals (Sweden)

    He Wang

    Full Text Available The pigmentation of many Aeromonas species has been thought to be due to the production of a L-DOPA (L-3,4-dihydroxyphenylalanine based melanin. However, in this study we found that although L-DOPA synthesis occurs in the high-melanin-yielding Aeromonas media strain WS, it plays a minor, if any, role in pigmentation. Instead, the pigmentation of A. media strain WS is due to the production of pyomelanin through HGA (homogentisate. Gene products of phhA (encodes phenylalanine hydroxylase, tyrB and aspC (both encode aromatic amino acid aminotransferase, and hppD (encodes 4-hydroxyphenylpyruvate dioxygenase constitute a linear pathway of converting phenylalanine to HGA and disruption of any one of these genes impairs or blocks pigmentation of A. media strain WS. This HGA biosynthesis pathway is widely distributed in Aeromonas, but HGA is only detectable in the cultures of pigmented Aeromonas species. Heterologous expression of HppD from both pigmented and non-pigmented Aeromonas species in E. coli leads to the production of pyomelanin and thus pigmentation, suggesting that most Aeromonas species have the critical enzymes to produce pyomelanin through HGA. Taken together, we have identified a widely conserved biosynthesis pathway of HGA based pyomelanin in Aeromonas that may be responsible for pigmentation of many Aeromonas species.

  7. Molecular detection and antimicrobial resistance of Aeromonas from houseflies (Musca domestica in Iran

    Directory of Open Access Journals (Sweden)

    Davood Ommi

    2015-11-01

    Full Text Available Objective. This study aimed to report the molecular detection and antimicrobial resistance of Aeromonas among houseflies (Musca domestica in Shahrekord and Isfahan provinces of Iran. Materials and methods. Flies were caught from household kitchens, cattle farms, animal hospitals, human hospitals, slaughter house and poultry farms and put in collection separate sterile tubes. Isolation was accomplished by culture of flies in alkaline peptone water followed by identification with Aeromonas-specific Polymerase Chain Reaction (PCR. Results. Out of 600 houseflies 73 (12.2% were infected with Aeromonas spp. Significantly higher frequencies of Aeromonas were isolated in Shahrekord province (13.0%; 39/300 than in Isfahan province (11.3%; 34/300. The recovery frequencies of the organisms were significantly lower in kitchens as compared to those in cattle farms and hospital wards which were similar. Higher proportions of infected flies were obtained during summer whereas low proportions were obtained during winter. Conclusions. It is concluded that houseflies do harbor diarrheagenic pathogens, including Aeromonas especially during summer. The carried organisms are resistant to a number of antimicrobials at different levels. Thus, future plans aimed at stemming infections caused by these organisms should take flies into account. Control efforts of infections caused by this particular bacterium should therefore take into account Musca domestica.

  8. Aeromonas spp. IN PRODUCTS OF ANIMAL ORIGIN AND DRINKING WATER OF GOIÂNIA-GO Aeromonas spp. EM PRODUTOS DE ORIGEM ANIMAL E EM ÁGUA DE CONSUMO DE GOIÂNIA - GO

    Directory of Open Access Journals (Sweden)

    Albenones José de Mesquita

    2007-09-01

    Full Text Available

    A total of 53 samples of products from animal origin and drinking water was examined for the presence of Aeromonas spp. Aeromonas occurred in 26.41% (14/53 of the samples. The raw products showed high frequency of contamination and relatively high count Aeromonas hydrophila was the dominating species, 57.5%, being followed of A. veroni (22.5%, A. sobria (17.5% and A. media (2.5%. None of the Aeromonas spp. was isolated from chlorinated or unchlorinated drinking water.

    KEY-WORDS: Aeromonas; animal origin products; drinking water.

    Um total de 53 amostras de produtos de origem animal e água de consumo foi examinado para se verificar a presença de Aeromonas spp. Desse total, 14/53 (26,41% amostras foram positivas para Aeromonas. Os produtos crus apresentaram maior freqüência de contaminação e contagem relativamente elevadas. A espécie Aeromonas hydrophila foi a mais freqüente (57,5%, seguida pelas espécies A. veroni (22,5%, A. sobria (17,5% e A. media (2,5%. Nenhuma amostra de água de consumo, clorada ou não clorada, mostrou-se positiva para Aeromonas ssp.

    PALAVRA-CHAVE: Aeromonas; produtos de origem animal; água de consumo.

  9. The chalcone butein from Rhus verniciflua Stokes inhibits clonogenic growth of human breast cancer cells co-cultured with fibroblasts

    Directory of Open Access Journals (Sweden)

    Tan Jenny

    2005-03-01

    Full Text Available Abstract Background Butein (3,4,2',4'-tetrahydroxychalone, a plant polyphenol, is a major biologically active component of the stems of Rhus verniciflua Stokes. It has long been used as a food additive in Korea and as an herbal medicine throughout Asia. Recently, butein has been shown to suppress the functions of fibroblasts. Because fibroblasts are believed to play an important role in promoting the growth of breast cancer cells, we investigated the ability of butein to inhibit the clonogenic growth of small numbers of breast cancer cells co-cultured with fibroblasts in vitro. Methods We first measured the clonogenic growth of small numbers of the UACC-812 human breast cancer cell line co-cultured on monolayers of serum-activated, human fibroblasts in the presence of butein (2 μg/mL or various other modulators of fibroblast function (troglitazone-1 μg/mL; GW9662-1 μM; meloxican-1 μM; and 3,4 dehydroproline-10 μg/mL. In a subsequent experiment, we measured the dose-response effect on the clonogenic growth of UACC-812 breast cancer cells by pre-incubating the fibroblasts with varying concentrations of butein (10 μg/ml-1.25 μg/mL. Finally, we measured the clonogenic growth of primary breast cancer cells obtained from 5 clinical specimens with normal fibroblasts and with fibroblasts that had been pre-treated with a fixed dose of butein (2.5 μg/mL. Results Of the five modulators of fibroblast function that we tested, butein was by far the most potent inhibitor of clonogenic growth of UACC-812 breast cancer cells co-cultured with fibroblasts. Pre-treatment of fibroblasts with concentrations of butein as low as 2.5 μg/mL nearly abolished subsequent clonogenic growth of UACC-812 breast cancer cells co-cultured with the fibroblasts. A similar dose of butein had no effect on the clonogenic growth of breast cancer cells cultured in the absence of fibroblasts. Significantly, clonogenic growth of the primary breast cancer cells was also

  10. Impact of the co-culture of Saccharomyces cerevisiae-Oenococcus oeni on malolactic fermentation and partial characterization of a yeast-derived inhibitory peptidic fraction.

    Science.gov (United States)

    Nehme, Nancy; Mathieu, Florence; Taillandier, Patricia

    2010-02-01

    The present study was aimed to evaluate the impact of the co-culture on the output of malolactic fermentation and to further investigate the reasons of the antagonism exerted by yeasts towards bacteria during sequential cultures. The Saccharomyces cerevisiae D strain/Oenococcus oeni X strain combination was tested by applying both sequential culture and co-culture strategies. This pair was chosen amongst others because the malolactic fermentation was particularly difficult to realize during the sequential culture. During this traditional procedure, malolactic fermentation started when alcoholic fermentation was achieved. For the co-culture, both fermentations were conducted together by inoculating yeasts and bacteria into a membrane bioreactor at the same time. Results obtained during the sequential culture and compared to a bacterial control medium, showed that the inhibition exerted by S. cerevisiae D strain in term of decrease of the malic acid consumption rate was mainly due to ethanol (75%) and to a peptidic fraction (25%) having an MW between 5 and 10 kDa. 0.4 g l(-1) of L-malic acid was consumed in this case while 3.7 g l(-1) was consumed when the co-culture was applied. In addition, there was no risk of increased volatile acidity during the co-culture. Therefore, the co-culture strategy was considered effective for malolactic fermentation with the yeast/bacteria pair studied.

  11. Characterization of chemical-induced sterile inflammation in vitro: application of the model compound ketoconazole in a human hepatic co-culture system.

    Science.gov (United States)

    Wewering, Franziska; Jouy, Florent; Wissenbach, Dirk K; Gebauer, Scarlett; Blüher, Matthias; Gebhardt, Rolf; Pirow, Ralph; von Bergen, Martin; Kalkhof, Stefan; Luch, Andreas; Zellmer, Sebastian

    2017-02-01

    Liver injury as a result of a sterile inflammation is closely linked to the activation of immune cells, including macrophages, by damaged hepatocytes. This interaction between immune cells and hepatocytes is as yet not considered in any of the in vitro test systems applied during the generation of new drugs. Here, we established and characterized a novel in vitro co-culture model with two human cell lines, HepG2 and differentiated THP-1. Ketoconazole, an antifungal drug known for its hepatotoxicity, was used as a model compound in the testing of the co-culture. Single cultures of HepG2 and THP-1 cells were studied as controls. Different metabolism patterns of ketoconazole were observed for the single and co-culture incubations as well as for the different cell types. The main metabolite N-deacetyl ketoconazole was found in cell pellets, but not in supernatants of cell cultures. Global proteome analysis showed that the NRF2-mediated stress response and the CXCL8 (IL-8) pathway were induced by ketoconazole treatment under co-culture conditions. The upregulation and ketoconazole-induced secretion of several pro-inflammatory cytokines, including CXCL8, TNF-α and CCL3, was observed in the co-culture system only, but not in single cell cultures. Taking together, we provide evidence that the co-culture model applied might be suitable to serve as tool for the prediction of chemical-induced sterile inflammation in liver tissue in vivo.

  12. A Preliminary Observation on the Development of Mouse Embryos Co-cultured with Human Oviductal Tissue or Conditioned Medium in Vitro

    Institute of Scientific and Technical Information of China (English)

    钟瑜; 张春雪; 潘善培

    1994-01-01

    The Present investigation has been carried out to examine the effect of human oviductal tissue co-culture system on the development of mouse embryos in vitro.Two-cell embryos collected from superovulated mouse were co-cultured with human oviductal tissue suspended in Ham'd F10+10%Fetal Calf Serum(F10 FCS),or in oviductal tissue conditioned medium and F10FCS as control.The results showed that the proportion developed into blastocyst,proportion of hatched and the velocity of cmbryo development were higher in both tissue co-culture and conditioned medium as compared with F10 FCS control.Furthermore,the velocity and percentage of embryomic devetopmem were higher in co-culture with ampullary tissue or its conditioned medium than that of isthmus,the effects of co-culture and conditioned medium on embryo development had no significant difference.All the embryos obtained from two co-culture systems could cleave normally,This experimental observation indicated that human oviductal epithelium might secrete some factors to promote the embryonic development in vitro.

  13. PINCH expression in relation to radiation response in co-cultured colon cancer cells and in rectal cancer patients.

    Science.gov (United States)

    Holmqvist, Annica; Holmlund, Birgitta; Ardsby, Malin; Pathak, Surajit; Sun, Xiao-Feng

    2013-11-01

    Particularly interesting new cysteine-histidine rich protein (PINCH), involved in cell spreading, motility and proliferation, has been shown to enhance radioresistance in colon cancer cell lines. The expression of PINCH in relation to radiation was studied in co-cultured colon cancer cells. Furthermore, the clinical significance between PINCH and radiotherapy (RT) was analyzed in rectal cancer patients with or without RT. The relative PINCH expression in colon cancer (KM12C) cells cultured separately and in co-culture was examined by western blotting and real-time PCR, and was analyzed over a period of 8 and 24 h after radiation. PINCH expression was immunohistochemically examined in 137 primary rectal tumors for which 65 cases did not receive RT and 72 cases received RT. PINCH expression tended to decrease from that in the separately cultured KM12C cells without radiation to that in cells with radiation at 8 h (P=0.060); while in the co-cultured cells, no significant difference was found (P=0.446). In patients with RT, strong PINCH expression was related to worse survival, when compared to patients with weak expression, independent of TNM stage, degree of differentiation, age and p53 status (P=0.029, RR 4.03, 95% CI 1.34‑12.1). No survival relationship for the patients without RT was observed (P=0.287). A statistical interaction analysis between PINCH, RT and survival showed a trend towards significance (P=0.057). In conclusion, PINCH predicts survival in rectal cancer patients with RT, but not in patients without RT. The expression of PINCH may be regulated by radiation and by environmental factors surrounding the cells.

  14. Effect of Bifidobacterium upon Clostridium difficile growth and toxicity when co-cultured in different prebiotic substrates

    Directory of Open Access Journals (Sweden)

    Lorena Valdés Varela

    2016-05-01

    Full Text Available The intestinal overgrowth of Clostridium difficile, often after disturbance of the gut microbiota by antibiotic treatment, leads to C. difficile infection (CDI which manifestation ranges from mild diarrhoea to life-threatening conditions. The increasing CDI incidence, not only in compromised subjects but also in traditionally considered low-risk populations, together with the frequent relapses of the disease, has attracted the interest for prevention/therapeutic options. Among these, probiotics, prebiotics or synbiotics constitute a promising approach. In this study we determined the potential of selected Bifidobacterium strains for the inhibition of C. difficile growth and toxicity in different carbon sources. We conducted co-cultures of the toxigenic strain C. difficile LMG21717 with four Bifidobacterium strains (Bifidobacterium longum IPLA20022, Bifidobacterium breve IPLA20006, Bifidobacterium bifidum IPLA20015, and Bifidobacterium animalis subsp. lactis Bb12 in the presence of various prebiotic substrates (Inulin, Synergy and Actilight or glucose, and compared the results with those obtained for the corresponding mono-cultures. C. difficile and bifidobacteria levels were quantified by qPCR; the pH and the production of short chain fatty acids was also determined. Moreover, supernatants of the cultures were collected to evaluate their toxicity using a recently developed model. Results showed that co-culture with B. longum IPLA20022 and B. breve IPLA20006 in the presence of short-chain fructooligosaccharides, but not of Inulin, as carbon source significantly reduced the growth of the pathogen. With the sole exception of B. animalis Bb12, whose growth was enhanced, the presence of C. difficile did not show major effects upon the growth of the bifidobacteria. In accordance with the growth data, B. longum and B. breve were the strains showing higher reduction in the toxicity of the co-culture supernatants.

  15. Development of a bovine luteal cell in vitro culture system suitable for co-culture with early embryos.

    Science.gov (United States)

    Batista, M; Torres, A; Diniz, P; Mateus, L; Lopes-da-Costa, L

    2012-10-01

    The cross talk between the corpus luteum (CL) and the early embryo, potentially relevant to pregnancy establishment, is difficult to evaluate in the in vivo bovine model. In vitro co-culture of bovine luteal cells and early embryos (days 2-8 post in vitro fertilization) may allow the deciphering of this poorly understood cross talk. However, early embryos and somatic cells require different in vitro culture conditions. The objective of this study was to develop a bovine luteal cell in vitro culture system suitable for co-culture with early embryos in order to evaluate their putative steroidogenic and prostanoid interactions. The corpora lutea of the different stages of the estrous cycle (early, mid, and late) were recovered postmortem and enriched luteal cell populations were obtained. In experiments 1 and 2, the effects of CL stage, culture medium (TCM, DMEM-F12, or SOF), serum concentration (5 or 10%), atmosphere oxygen tension (5 or 20%), and refreshment of the medium on the ability of luteal cells to produce progesterone (P(4)) were evaluated. The production of P(4) was significantly increased in early CL cultures, and luteal cells adapted well to simple media (SOF), low serum concentrations (5%), and oxygen tensions (5%). In experiment 3, previous luteal cell cryopreservation did not affect the production of P(4), PGF(2α), and PGE(2) compared to fresh cell cultures. This enables the use of pools of frozen-thawed cells to decrease the variation in cell function associated with primary cell cultures. In experiment 4, mineral oil overlaying culture wells resulted in a 50-fold decrease of the P(4) quantified in the medium, but had no effect on PGF(2α) and PGE(2) quantification. In conclusion, a luteal cell in vitro culture system suitable for the 5-d-long co-culture with early embryos was developed.

  16. Modeling long-term host cell-Giardia lamblia interactions in an in vitro co-culture system.

    Science.gov (United States)

    Fisher, Bridget S; Estraño, Carlos E; Cole, Judith A

    2013-01-01

    Globally, there are greater than 700,000 deaths per year associated with diarrheal disease. The flagellated intestinal parasite, Giardia lamblia, is one of the most common intestinal pathogens in both humans and animals throughout the world. While attached to the gastrointestinal epithelium, Giardia induces epithelial cell apoptosis, disrupts tight junctions, and increases intestinal permeability. The underlying cellular and molecular mechanisms of giardiasis, including the role lamina propria immune cells, such as macrophages, play in parasite control or clearance are poorly understood. Thus far, one of the major obstacles in ascertaining the mechanisms of Giardia pathology is the lack of a functionally relevant model for the long-term study of the parasite in vitro. Here we report on the development of an in vitro co-culture model which maintains the basolateral-apical architecture of the small intestine and allows for long-term survival of the parasite. Using transwell inserts, Caco-2 intestinal epithelial cells and IC-21 macrophages are co-cultured in the presence of Giardia trophozoites. Using the developed model, we show that Giardia trophozoites survive over 21 days and proliferate in a combination media of Caco-2 cell and Giardia medium. Giardia induces apoptosis of epithelial cells through caspase-3 activation and macrophages do not abrogate this response. Additionally, macrophages induce Caco-2 cells to secrete the pro-inflammatory cytokines, GRO and IL-8, a response abolished by Giardia indicating parasite induced suppression of the host immune response. The co-culture model provides additional complexity and information when compared to a single-cell model. This model will be a valuable tool for answering long-standing questions on host-parasite biology that may lead to discovery of new therapeutic interventions.

  17. Modeling long-term host cell-Giardia lamblia interactions in an in vitro co-culture system.

    Directory of Open Access Journals (Sweden)

    Bridget S Fisher

    Full Text Available Globally, there are greater than 700,000 deaths per year associated with diarrheal disease. The flagellated intestinal parasite, Giardia lamblia, is one of the most common intestinal pathogens in both humans and animals throughout the world. While attached to the gastrointestinal epithelium, Giardia induces epithelial cell apoptosis, disrupts tight junctions, and increases intestinal permeability. The underlying cellular and molecular mechanisms of giardiasis, including the role lamina propria immune cells, such as macrophages, play in parasite control or clearance are poorly understood. Thus far, one of the major obstacles in ascertaining the mechanisms of Giardia pathology is the lack of a functionally relevant model for the long-term study of the parasite in vitro. Here we report on the development of an in vitro co-culture model which maintains the basolateral-apical architecture of the small intestine and allows for long-term survival of the parasite. Using transwell inserts, Caco-2 intestinal epithelial cells and IC-21 macrophages are co-cultured in the presence of Giardia trophozoites. Using the developed model, we show that Giardia trophozoites survive over 21 days and proliferate in a combination media of Caco-2 cell and Giardia medium. Giardia induces apoptosis of epithelial cells through caspase-3 activation and macrophages do not abrogate this response. Additionally, macrophages induce Caco-2 cells to secrete the pro-inflammatory cytokines, GRO and IL-8, a response abolished by Giardia indicating parasite induced suppression of the host immune response. The co-culture model provides additional complexity and information when compared to a single-cell model. This model will be a valuable tool for answering long-standing questions on host-parasite biology that may lead to discovery of new therapeutic interventions.

  18. Cell differentiation mediated by co-culture of human umbilical cord blood stem cells with murine hepatic cells.

    Science.gov (United States)

    Stecklum, Maria; Wulf-Goldenberg, Annika; Purfürst, Bettina; Siegert, Antje; Keil, Marlen; Eckert, Klaus; Fichtner, Iduna

    2015-02-01

    In the present study, purified human cord blood stem cells were co-cultivated with murine hepatic alpha mouse liver 12 (AML12) cells to compare the effect on endodermal stem cell differentiation by either direct cell-cell interaction or by soluble factors in conditioned hepatic cell medium. With that approach, we want to mimic in vitro the situation of preclinical transplantation experiments using human cells in mice. Cord blood stem cells, cultivated with hepatic conditioned medium, showed a low endodermal differentiation but an increased connexin 32 (Cx32) and Cx43, and cytokeratin 8 (CK8) and CK19 expression was monitored by reverse transcription polymerase chain reaction (RT-PCR). Microarray profiling indicated that in cultivated cord blood cells, 604 genes were upregulated 2-fold, with the highest expression for epithelial CK19 and epithelial cadherin (E-cadherin). On ultrastructural level, there were no major changes in the cellular morphology, except a higher presence of phago(ly)some-like structures observed. Direct co-culture of AML12 cells with cord blood cells led to less incisive differentiation with increased sex-determining region Y-box 17 (SOX17), Cx32 and Cx43, as well as epithelial CK8 and CK19 expressions. On ultrastructural level, tight cell contacts along the plasma membranes were revealed. FACS analysis in co-cultivated cells quantified dye exchange on low level, as also proved by time relapse video-imaging of labelled cells. Modulators of gap junction formation influenced dye transfer between the co-cultured cells, whereby retinoic acid increased and 3-heptanol reduced the dye transfer. The study indicated that the cell-co-cultured model of human umbilical cord blood cells and murine AML12 cells may be a suitable approach to study some aspects of endodermal/hepatic cell differentiation induction.

  19. POTENSI TUMBUHAN BANGKAL (Nauclea Orientalis UNTUK PENGENDALIAN BAKTERI Aeromonas Hydrophila (POTENTIAL PLANT BANGKAL (Nauclea orientalis FOR CONTROL Aeromonas Hydrophila

    Directory of Open Access Journals (Sweden)

    Siti Aisiah

    2017-02-01

    Full Text Available Tujuan penelitian ini adalah didapatkan metode pengendalian penyakit yang disebabkan oleh bakteri Aeromonas hydrophila pada budi daya ikan yang ramah lingkungan. Dalam penelitian ini dilakukan uji sensitivitas terhadap bakteri A. hydrophila, uji minimal konsentrasi menghambat  bakteri A. hydrophila (uji MIC dan uji toksisitas terhadap ikan nila. Rancangan  yang digunakan untuk  uji toksisitas  adalah rancangan acak lengkap (RAL, dengan 4 perlakuan yaitu A  = Ikan disuntik dengan ekstrak bangkal konsentrasi 20%, B  = Ikan disuntik dengan ekstrak bangkal konsentrasi 40%,  C  = Ikan disuntik dengan ekstrak bangkal konsentrasi 80%, dan D  = Kontrol (ikan tidak disuntik, diulang sebanyak 3 kali. Perlakuan ini didapat dari hasil uji sensitivitas antibakteri bangkal yang mempunyai daya hambat dan daya bunuh paling besar terhadap bakteri A. hydrophila yaitu ekstrak daun bangkal dengan pelarut akuades. Pengujian MIC menunjukkan bahwa ekstrak bangkal-akuades memiliki daya hambat minimal  20 % terhadap aktivitas bakteri A. hydrophila. Hasil uji toksisitas yang dilakukan terhadap ikan nila dengan konsentrasi 20%, 40% dan 80%  mortalitas ikan nila dibawah 50 %. Pengamatan hematologis yaitu eretrosit, leokosit, plasma darah, hematokrit dan leokokrit pada masing-masing perlakuan sebagian besar masih berada dalam kisaran yang normal. Parameter kualitas air yaitu, kadar oksigen terlarut,  pH, amoniak, CO2 dan suhu masih dapat mendukung kehidupan normal ikan nila. The purpose of this study was obtained method of controlling diseases caused by Aeromonas hydrophila in fish farming is environmentally friendly. In this study the sensitivity of the test  A. hydrophila, a minimum test konsentari inhibiting A. hydrophila (MIC test and toxicity test on  tilapia. The design used for toxicity tests is completely randomized design with 4 treatments it A = Fish injected with extracts bangkal concentration of 20 %,  B = Fish injected with extracts bangkal

  20. Physiological control of amonabactin biosynthesis in Aeromonas hydrophila.

    Science.gov (United States)

    Barghouthi, S; Young, R; Arceneaux, J E; Byers, B R

    1989-01-01

    Amonabactin is a siderophore from Aeromonas hydrophila which is produced in two biologically active forms composed of the phenolate 2,3-dihydroxybenzoic acid (DHB), lysine, glycine, and either trytophan (amonabactin T) or phenylalanine (amonabactin P). Amonabactin biosynthetic mutants (generated by chemical mutagenesis) that either produced no amonabactin or overproduced the siderophore were isolated and identified on chrome azurol S siderophore detection agar. Amonabactin-negative mutants were of two categories. One type produced no phenolates and used exogenous DHB to synthesize amonabactin (both forms) while the other type excreted DHB but not amonabactin. This suggests an amonabactin biosynthetic pathway composed of two segments, one producing DHB and the other assembling amonabactin from DHB and the amino acids. Overproduction mutants used amonabactin poorly or not at all, indicating that they contained lesions in amonabactin utilization. Adding the analog D-tryptophan to wild-type A. hydrophila cultures reduced synthesis of both amonabactin T and amonabactin P and lengthened the lag phase in iron restricted medium. The tryptophan and phenylalanine forms of amonabactin may be synthesized by a single assembly pathway that contains a novel enzyme (sensitive to D-tryptophan) which inserts either tryptophan or phenylalanine into amonabactin.

  1. Enterobacter asburiae and Aeromonas hydrophila: soft tissue infection requiring debridement.

    Science.gov (United States)

    Koth, Kevin; Boniface, James; Chance, Elisha A; Hanes, Marina C

    2012-06-01

    Enterobacter asburiae and Aeromonas hydrophila are gram-negative bacilli that have been isolated in soil and water. Enterobacter asburiae can cause an array of diseases, and exposure to A hydrophila can cause soft tissue infections, including necrotizing faciitis.A healthy-appearing 22-year-old man presented with an innocuous soft tissue injury to his leg due to an all-terrain vehicle crash. He received intravenous antibiotics and was discharged with prophylactic oral antibiotics. After the rapid onset of high fevers (102°F-103°F) <24 hours postinjury, he returned to the emergency department. Emergent surgical debridement was performed, and broad-spectrum intravenous antibiotics were started. Fevers persisted, and the patient underwent repeat extensive surgical debridement and antibiotic bead placement <30 hours after the initial surgical debridement and broad-spectrum antibiotics. Intraoperative cultures found E asburiae and A hydrophila in the wound. Following a long course of antibiotics and a skin graft, he fully recovered and had no functional deficits 1 year postoperatively.Extensive research revealed that these organisms are rare in soft tissue infections. E asburiae is opportunistic but has not been reported as a primary wound organism, and A hydrophila infections have been reported following motor vehicle crashes involving wound contamination. At presentation, it is challenging to determine rare organisms in a timely fashion; however, emergent extensive surgical intervention of an accelerated aberrant disease process should be considered to avoid catastrophic outcomes.

  2. 中国豹源豚鼠气单胞菌的分子鉴定与毒力基因分析%Molecular Identification and Virulent Gene Analysis of Aeromonas Caviae from China Leopard(Panthera pardus japonensis)

    Institute of Scientific and Technical Information of China (English)

    李婉萍; 彭仕明; 陈武; 胡俊; 李少基; 单芬

    2015-01-01

    从表现为急性出血性败血症的中国豹分离到一株细菌,为了鉴定这株细菌,并对其主要毒力基因进行检测分析,实验中进行了细菌的分离培养、生化鉴定及动物毒力试验;PCR扩增gyrB基因并结合系统发育树的构建和分析进行了菌种的分子鉴定;对分离菌进行了气溶素(aerolysin,aer)、热敏感细胞肠毒素(heat-Labile cytotonic enterotoxin,alt)、细胞毒性肠毒素(cytotoxic enterotoxin,act)、热敏蛋白酶(temperature-sensitive protease,eprCAI)、丝氨酸蛋白酶(serine protease,ahp)5种毒力基因的PCR扩增和测序.结果显示:根据培养特性、生化试验、小白鼠毒力试验鉴定分离菌为豚鼠气单胞菌;中国豹源细菌gyrB基因与豚鼠气单胞菌(JQ815381)的同源性为98.9%,应为豚鼠气单胞菌;具有act、alt、eprACI 3种毒力基因.该研究可为快捷、准确检测气单胞菌引起的疾病提供参考.

  3. Tooth length and incisal wear and growth in guinea pigs (Cavia porcellus) fed diets of different abrasiveness.

    Science.gov (United States)

    Müller, J; Clauss, M; Codron, D; Schulz, E; Hummel, J; Kircher, P; Hatt, J-M

    2015-06-01

    Dental diseases are among the most important reasons for presenting guinea pigs (Cavia porcellus) and other rodents to veterinary clinics, but the aetiopathology of this disease complex is unclear. Clinicians tend to believe that the ever-growing teeth of rabbits and rodents have a constant growth that needs to be worn down by the mastication of an appropriate diet. In this study, we tested the effect of four different pelleted diets of increasing abrasiveness [due to both internal (phytoliths) and external abrasives (sand)] or whole grass hay fed for 2 weeks each in random order to 16 guinea pigs on incisor growth and wear, and tooth length of incisors and cheek teeth. There was a positive correlation between wear and growth of incisors. Tooth lengths depended both on internal and external abrasives, but only upper incisors were additionally affected by the feeding of whole hay. Diet effects were most prominent in anterior cheek teeth, in particular M1 and m1. Cheek tooth angle did not become shallower with decreasing diet abrasiveness, suggesting that a lack of dietary abrasiveness does not cause the typical 'bridge formation' of anterior cheek teeth frequently observed in guinea pigs. The findings suggest that other factors than diet abrasiveness, such as mineral imbalances and in particular hereditary malocclusion, are more likely causes for dental problems observed in this species. Journal of Animal Physiology and Animal Nutrition © 2014 Blackwell Verlag GmbH.

  4. Hyperendemic Campylobacter jejuni in guinea pigs (Cavia porcellus) raised for food in a semi-rural community of Quito, Ecuador.

    Science.gov (United States)

    Graham, Jay P; Vasco, Karla; Trueba, Gabriel

    2016-06-01

    Domestic animals and animal products are the source of pathogenic Campylobacter jejuni and C. coli in industrialized countries, yet little is known about the transmission of these bacteria in developing countries. Guinea pigs (Cavia porcellus) are commonly raised for food in the Andean region of South America, however, limited research has characterized this rodent as a reservoir of zoonotic enteric pathogens. In this study, we examined the prevalence of Campylobacter spp. in 203 fecal samples from domestic animals of 59 households in a semi-rural parish of Quito, Ecuador. Of the twelve animal species studied, guinea pigs showed the highest prevalence of C. jejuni (n = 39/40; 97.5%). Multilocus sequence typing (MLST) was used to characterize the genetic relationship of C. jejuni from domestic animals and 21 sequence types (STs) were identified. The majority of STs from guinea pigs appeared to form new clonal complexes that were not related to STs of C. jejuni isolated from other animal species and shared only a few alleles with other C. jejuni previously characterized. The study identifies guinea pigs as a major reservoir of C. jejuni and suggests that some C. jejuni strains are adapted to this animal species. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  5. Coagulation and fibrinolysis in capybara (Hydrochaeris hydrochaeris), a close relative of the guinea-pig (Cavia porcellus).

    Science.gov (United States)

    Leitão, D P; Polizello, A C; Rothschild, Z

    2000-01-01

    Fibrinolytic and coagulation properties of capybara (Hydrochaeris hydrochaeris, LINNAEUS, 1766) plasma were analysed and the results compared to the guinea-pig (Cavia porcellus), a close relative. Capybara fibrinogen was isolated and fibrinolysis of its plasma was carried out in a homologous system and with bovine fibrin. Undiluted plasma did not have fibrinolytic activity on fibrin plates; euglobulins gave a dose-related response. Zymography of capybara and guinea-pig plasma gave the same patterns of activity as human or bovine plasma. Human urokinase (UK) and tissue plasminogen activator (t-PA) produced lysis in capybara fibrin plates. Streptokinase (SK) (500 IU/ml) did not activate capybara or guinea-pig plasma. In this system, human plasma was extensively activated. Coagulation tests for both species of rodent were prolonged. The capybara showed values for prothrombin time (PT) shorter than activated thromboplastin time (APTT). The guinea-pig, as already shown, had longer PT values. Factors X and VII were very low for capybara and guinea-pig when tested using reference curves and diagnostic kits for human plasma. It is suggested that the capybara could be a valuable laboratory animal considering its size and closeness to the guinea-pig, and this could allow for the provision of materials from one single animal when convenient or necessary.

  6. Evaluation of the infectivity and the persistence of Trichinella patagoniensis in muscle tissue of decomposing guinea pig (Cavia porcellus).

    Science.gov (United States)

    Fariña, F; Pasqualetti, M; Ilgová, J; Cardillo, N; Ercole, M; Aronowicz, T; Krivokapich, S; Kašný, M; Ribicich, M

    2017-01-01

    Trichinella patagoniensis, a new species of Trichinella, is widespread in Argentina. The success of parasite transmission depends, among other factors, on the resistance of L1 larvae present in the muscle tissue (ML) of dead hosts undergoing the decomposition process in different environmental conditions. The aim of the present work was to study the infectivity of T. patagoniensis muscle larvae in Cavia porcellus and the capability of the parasite to survive in decomposed muscle tissue of guinea pigs subjected to different environmental conditions. Thirty-two female Ssi:AL guinea pigs were orally inoculated with 2000 ML of T. patagoniensis (ISS2311). All the animals were sacrificed 42 days post-infection. Twenty-six animals were eviscerated, and carcasses were placed on the surface of soil inside plastic boxes that were exposed to environmental conditions in the summer 2014-2015 and autumn of 2015 in Buenos Aires, Argentina. Carcasses from six animals were placed into a plastic box inside the refrigerator at a temperature of 4 °C. The muscle tissue samples from the carcasses were examined weekly for the presence of larvae, and the infectivity of recovered ML was tested in BALB/c mice. Our results showed for the first time the ability of T. patagoniensis to complete its life cycle in guinea pigs, thus serving as a potential natural host. Also, larvae of T. patagoniensis remained infective in muscle tissue for several weeks while undergoing decomposition under different environmental conditions.

  7. Hyperendemic Campylobacter jejuni in guinea pigs (Cavia porcellus) raised for food in a semi-rural community of Quito, Ecuador

    Science.gov (United States)

    Graham, Jay P.; Vasco, Karla; Trueba, Gabriel

    2016-01-01

    Summary Domestic animals and animal products are the source of pathogenic Campylobacter jejuni and C. coli in industrialized countries, yet little is known about the transmission of these bacteria in developing countries. Guinea pigs (Cavia porcellus) are commonly raised for food in the Andean region of South America, however, limited research has characterized this rodent as a reservoir of zoonotic enteric pathogens. In this study, we examined the prevalence of Campylobacter spp. in 203 fecal samples from domestic animals of 59 households in a semi-rural parish of Quito, Ecuador. Of the twelve animal species studied, guinea pigs showed the highest prevalence of C. jejuni (n = 39/40; 97.5%). Multilocus sequence typing (MLST) was used to characterize the genetic relationship of C. jejuni from domestic animals and 21 sequence types (STs) were identified. The majority of STs from guinea pigs appeared to form new clonal complexes that were not related to STs of C. jejuni isolated from other animal species and shared only a few alleles with other C. jejuni previously characterized. The study identifies guinea pigs as a major reservoir of C. jejuni and suggests that some C. jejuni strains are adapted to this animal species. PMID:27043446

  8. Evaluating the clinical and physiological effects of long term ultraviolet B radiation on guinea pigs (Cavia porcellus).

    Science.gov (United States)

    Watson, Megan K; Stern, Adam W; Labelle, Amber L; Joslyn, Stephen; Fan, Timothy M; Leister, Katie; Kohles, Micah; Marshall, Kemba; Mitchell, Mark A

    2014-01-01

    Vitamin D is an important hormone in vertebrates. Most animals acquire this hormone through their diet, secondary to exposure to ultraviolet B (UVB) radiation, or a combination thereof. The objectives for this research were to evaluate the clinical and physiologic effects of artificial UVB light supplementation on guinea pigs (Cavia porcellus) and to evaluate the long-term safety of artificial UVB light supplementation over the course of six months. Twelve juvenile acromelanic Hartley guinea pigs were randomly assigned to one of two treatment groups: Group A was exposed to 12 hours of artificial UVB radiation daily and Group B received only ambient fluorescent light for 12 hours daily. Animals in both groups were offered the same diet and housed under the same conditions. Blood samples were collected every three weeks to measure blood chemistry values, parathyroid hormone, ionized calcium, and serum 25-hydroxyvitamin D3 (25-OHD3) levels. Serial ophthalmologic examinations, computed tomography scans, and dual energy x-ray absorptiometry scans were performed during the course of the study. At the end of the study the animals were euthanized and necropsied. Mean ± SD serum 25-OHD3 concentrations differed significantly in the guinea pigs (pguinea pigs to UVB radiation long term significantly increased their circulating serum 25-OHD3 levels, and that this increase was sustainable over time. Providing guinea pigs exposure to UVB may be an important husbandry consideration that is not currently recommended.

  9. A new cyclopeptide with antifungal activity from the co-culture broth of two marine mangrove fungi.

    Science.gov (United States)

    Li, Chunyuan; Wang, Jinhua; Luo, Cuiping; Ding, Weijia; Cox, Daniel G

    2014-01-01

    A new cyclic tetrapeptide, cyclo-(L-leucyl-trans-4-hydroxy-L-prolyl-D-leucyl-trans-4-hydroxy-L-proline) (1), was isolated from the co-culture broth of two mangrove fungi Phomopsis sp. K38 and Alternaria sp. E33. The structure of 1 was determined by analysis of spectroscopic data and Marfey's analytic method. Primary bioassay demonstrated that compound 1 exhibited moderate to high inhibitory activity against four crop-threatening fungi including Gaeumannomyces graminis, Rhizoctonia cerealis, Helminthosporium sativum and Fusarium graminearum as compared with triadimefon.

  10. Co-culture fermentation of peanut-soy milk for the development of a novel functional beverage.

    Science.gov (United States)

    Santos, Claudia Cristina Auler do Amaral; Libeck, Bárbara da Silva; Schwan, Rosane Freitas

    2014-09-01

    Most of the commercial probiotic products are dairy-based, and the development of non-dairy probiotic products could be an alternative for new functional products. The peanut-soy milk (PSM(1)) was inoculated with six different lactic acid bacteria (LAB), including probiotic strains and yeasts and fermentation was accomplished for 24h at 37 °C and afterwards, another 24h at ±4 °C. The Pediococcus acidilactici (UFLA BFFCX 27.1), Lactococcus lactis (CCT 0360), Lactobacillus rhamnosus (LR 32) probiotic LAB, and the Lactobacillus delbrueckii subsp. bulgaricus (LB 340) yogurt starter culture reached cell concentrations of about 8.3log CFU/mL during fermentation. However, these strains were not able to acidify the substrate when inoculated as pure culture. The Lactobacillus acidophilus (LACA 4) probiotic produced significant amounts of lactic acid (3.35 g/L) and rapidly lowered the pH (4.6). Saccharomyces cerevisiae (UFLA YFFBM 18.03) did not completely consume the available sugars in PSM and consequently produced low amounts of ethanol (0.24 g/L). In pure culture, S. cerevisiae (UFLA YFFBM 18.03), L. rhamnosus (LR 32), L. acidophilus (LACA 4), and P. acidilactici (UFLA BFFCX 27.1) promoted the increase of total amino acids (48.02%, 47.32%, 46.21% and, 44.07%, respectively). However, when in co-cultured, the strains consumed the free amino acids favoring their growth, and reaching the population of 8log CFU/mL in PSM. Lactic acid production increased, and 12 h was required to reach a pH value of 4.3. In general, the strains were more efficient in the use of available carbohydrates and release of metabolites in co-cultured than in single culture fermentations. An average of 58% and 78% of available carbohydrates was consumed when single and co-cultures were evaluated, respectively. Higher lactic acid contents were found in a binary culture of P. acidilactici (UFLA BFFCX 27.1) and L. acidophilus (LACA 4), and by co-culture of P. acidilactici (UFLA BFFCX 27.1), L

  11. Three-Dimensional Organotypic Co-Culture Model of Intestinal Epithelial Cells and Macrophages to Study "Salmonella Enterica" Colonization Patterns

    Science.gov (United States)

    Ott, Mark; Yang, J; Barilla, J.; Crabbe, A.; Sarker, S. F.; Liu, Y.

    2017-01-01

    Three-dimensional/3-D organotypic models of human intestinal epithelium mimic the differentiated form and function of parental tissues often not exhibited by 2-D monolayers and respond to Salmonella in ways that reflect in vivo infections. To further enhance the physiological relevance of 3-D models to more closely approximate in vivo intestinal microenvironments during infection, we developed and validated a novel 3-D intestinal co-culture model containing multiple epithelial cell types and phagocytic macrophages, and applied to study enteric infection by different Salmonella pathovars.

  12. Aeromonas cavernicola sp. nov., isolated from fresh water of a brook in a cavern.

    Science.gov (United States)

    Martínez-Murcia, Antonio; Beaz-Hidalgo, Roxana; Svec, Pavel; Saavedra, Ma José; Figueras, Ma José; Sedlacek, Ivo

    2013-02-01

    Aeromonas P2973 was isolated from the water of a brook in a cavern in the Czech Republic. This isolate could not be biochemically identified at the species level, considering all updated species descriptions. Subsequent extensive phenotypic characterisation, DNA-DNA hybridisation, 16S rRNA gene sequencing and a Multi-Locus Phylogenetic Analysis (MLPA) of the concatenated sequence of 7 housekeeping genes (gyrB, rpoD, recA, dnaJ, gyrA, dnaX and atpD; 4705 bp) was employed in an attempt to ascertain the taxonomy of this isolate. Based on this polyphasic approach, we describe a novel species of the genus Aeromonas, for which the name Aeromonas cavernicola sp. nov. is proposed, with strain CCM7641(T) (DSM24474(T), CECT7862(T)) as the type strain.

  13. Evaluation of the MALDI-TOF MS profiling for identification of newly described Aeromonas spp.

    Science.gov (United States)

    Vávrová, Andrea; Balážová, Tereza; Sedláček, Ivo; Tvrzová, Ludmila; Šedo, Ondrej

    2015-09-01

    The genus Aeromonas comprises primarily aquatic bacteria and also serious human and animal pathogens with the occurrence in clinical material, drinking water, and food. Aeromonads are typical for their complex taxonomy and nomenclature and for limited possibilities of identification to the species level. According to studies describing the use of MALDI-TOF MS in diagnostics of aeromonads, this modern chemotaxonomical approach reveals quite high percentage of correctly identified isolates. We analyzed 64 Aeromonas reference strains from the set of 27 species. After extending the range of analyzed Aeromonas species by newly described ones, we proved that MALDI-TOF MS procedure accompanied by Biotyper tool is not a reliable diagnostic technique for aeromonads. We obtained quite high percentage of false-positive, incorrect, and uncertain results. The identification of newly described species is accompanied with misidentifications that were observed also in the case of pathogenic aeromonads.

  14. In vitro differentiation of bone marrow stromal cells into neurons and glial cells and differential protein expression in a two-compartment bone marrow stromal cell/neuron co-culture system.

    Science.gov (United States)

    Qi, Xu; Shao, Ming; Peng, Haisheng; Bi, Zhenggang; Su, Zhiqiang; Li, Hulun

    2010-07-01

    This study was performed to establish a bone marrow stromal cell (BMSC)/neuron two-compartment co-culture model in which differentiation of BMSCs into neurons could occur without direct contact between the two cell types, and to investigate protein expression changes during differentiation of this entirely BMSC-derived population. Cultured BMSCs isolated from Wistar rats were divided into three groups: BMSC culture, BMSC/neuron co-culture and BMSC/neuron two-compartment co-culture. Cells were examined for neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression. The electrophysiological behavior of the BMSCs was examined using patch clamping. Proteins that had significantly different expression levels in BMSCs cultured alone and co-cultured with neurons were studied using a protein chip-mass spectroscopy technique. Expression of NSE and GFAP were significantly higher in co-culture cells than in two-compartment co-culture cells, and significantly higher in both co-culture groups than in BMSCs cultured alone. Five proteins showed significant changes in expression during differentiation: TIP39_RAT and CALC_RAT underwent increases, and INSL6_RAT, PNOC_RAT and PCSK1_RAT underwent decreases in expression. We conclude that BMSCs can differentiate into neurons during both contact co-culture with neurons and two-compartment co-culture with neurons. The rate at which BMSCs differentiated into neurons was higher in contact co-culture than in non-contact co-culture.

  15. Phylogenetic diversity, antibiotic resistance and virulence traits of Aeromonas spp. from untreated waters for human consumption.

    Science.gov (United States)

    Carvalho, Maria João; Martínez-Murcia, Antonio; Esteves, Ana Cristina; Correia, António; Saavedra, Maria José

    2012-10-15

    It is well known that water constitutes an important contamination route for microorganisms. This is especially true for Aeromonas which are widespread in untreated and treated waters. In this study, Portuguese untreated waters not regularly monitored were screened for the presence and diversity of aeromonads. A total of 206 isolates were discriminated by RAPD-PCR and 80 distinct strains were identified by gyrB based phylogenetic analysis. The most frequently detected species were Aeromonas hydrophila, Aeromonas bestiarum and Aeromonas media. The antibiotic susceptibility profile of these strains was determined and showed a typical profile of the genus. Nonetheless, the percentage of resistant strains to tetracycline, chloramphenicol and/or trimethoprim/sulfamethoxazole was lower than that reported for clinical isolates and isolates recovered from aquacultures and other environments historically subjected to antibiotic contamination. This suggests that the existence of such pressures in those environments selects for resistant Aeromonas. A similar trend for integron presence was found. Genes coding for CphA and TEM, and tet(A), (E), (C) or (D) genes were found in 28%, 1%, and 10% of the strains, respectively. 10% of the strains contained an integron. Variable regions of seven class 1 integrons and one class 2 integron were characterised. Furthermore, strains displayed virulence related phenotypes such as extracellular lipolytic and proteolytic activities as well as aerolysin related genes (43% of strains). The ascV and aexT genes were found in 16% and 3% of strains respectively and, in some cases, concomitantly in the same specimen. This study shows that diverse Aeromonas spp. presenting distinct antibiotic resistance features and putative virulence traits are frequently present in waters for human and animal consumption in Portugal. Genes associated to antibiotic resistance and microbial virulence previously identified in organisms with human health significance

  16. Co-culture of dedifferentiated and primary human chondrocytes obtained from cadaveric donor enhance the histological quality of repair tissue: an in-vivo animal study.

    Science.gov (United States)

    Olivos-Meza, Anell; Velasquillo Martínez, Cristina; Olivos Díaz, Brenda; Landa-Solís, Carlos; Brittberg, Mats; Pichardo Bahena, Raul; Ortega Sanchez, Carmina; Martínez, Valentin; Alvarez Lara, Enrique; Ibarra-Ponce de León, José Clemente

    2017-06-05

    To compare the quality of the repair tissue in three-dimensional co-culture of human chondrocytes implanted in an in vivo model. Six cadaveric and five live human donors were included. Osteochondral biopsies from the donor knees were harvested for chondrocyte isolation. Fifty percent of cadaveric chondrocytes were expanded until passage-2 (P2) while the remaining cells were cryopreserved in passage-0 (P0). Fresh primary chondrocytes (P0f) obtained from live human donors were co-cultured. Three-dimensional constructs were prepared with a monolayer of passage-2 chondrocytes, collagen membrane (Geistlich Bio-Gide(®)), and pellet of non-co-cultured (P2) or co-cultured chondrocytes (P2 + P0c, P2 + P0f). Constructs were implanted in the subcutaneous tissue of athymic mice and left for 3 months growth. Safranin-O and Alcian blue staining were used to glycosaminoglycan content assessment. Aggrecan and type-II collagen were evaluated by immunohistochemistry. New-formed tissue quality was evaluated with an adaptation of the modified O'Driscoll score. Histological quality of non-co-cultured group was 4.37 (SD ±4.71), while co-cultured groups had a mean score of 8.71 (SD ±3.98) for the fresh primary chondrocytes and 9.57 (SD ±1.27) in the cryopreserved chondrocytes. In immunohistochemistry, Co-culture groups were strongly stained for type-II and aggrecan not seen in the non-co-cultured group. It is possible to isolate viable chondrocytes from cadaveric human donors in samples processed in the first 48-h of dead. There is non-significant difference between the numbers of chondrocytes isolated from live or cadaveric donors. Cryopreservation of cadaveric primary chondrocytes does not alter the capability to form cartilage like tissue. Co-culture of primary and passaged chondrocytes enhances the histological quality of new-formed tissue compared to non-co-cultured cells.

  17. Fiber degradation potential of natural co-cultures of Neocallimastix frontalis and Methanobrevibacter ruminantium isolated from yaks (Bos grunniens) grazing on the Qinghai Tibetan Plateau.

    Science.gov (United States)

    Wei, Ya-Qin; Long, Rui-Jun; Yang, Hui; Yang, Hong-Jian; Shen, Xi-Hui; Shi, Rui-Fang; Wang, Zhi-Ye; Du, Jun-Guo; Qi, Xiao-Jin; Ye, Qian-Hong

    2016-06-01

    Several natural anaerobic fungus-methanogen co-cultures have been isolated from rumen and feces source of herbivores with strong fiber degrading ability. In this study, we isolated 7 Neocallimastix with methanogen co-cultures from the rumen of yaks grazing on the Qinghai Tibetan Plateau. Based on morphological characteristics and internal transcribed spacer 1 sequences (ITS1), all the fungi were identified as Neocallimastix frontalis. The co-cultures were confirmed as the one fungus - one methanogen pattern by the PCR-denatured gradient gel electrophoresis (DGGE) assay. All the methanogens were identified as Methanobrevibacter ruminantium by 16s rRNA gene sequencing. We investigated the biodegrading capacity of the co-culture (N. frontalis + M. ruminantium) Yaktz1 on wheat straw, corn stalk and rice straw in a 7 days-incubation. The in vitro dry matter digestibility (IVDMD), acid detergent fiber digestibility (ADFD) and neural detergent fiber digestibility (NDFD) values of the substrates in the co-culture were significantly higher than those in the mono-culture N. frontalis Yaktz1. The co-culture exhibited high polysaccharide hydrolase (xylanase and FPase) and esterase activities. The xylanase in the co-culture reached the highest activity of 12500 mU/ml on wheat straw at the day 3 of the incubation. At the end of the incubation, 3.00 mmol-3.29 mmol/g dry matter of methane were produced by the co-culture. The co-culture also produced high level of acetate (40.00 mM-45.98 mM) as the end-product during the biodegradation. Interestingly, the N. frontalis Yaktz1 mono-culture produced large amount of lactate (8.27 mM-11.60 mM) and ethanol (163.11 mM-242.14 mM), many times more than those recorded in the previously reported anaerobic fungi. Our data suggests that the (N. frontalis + M. ruminantium) Yaktz1 co-culture and the N. frontalis Yaktz1 mono-culture both have great potentials for different industrial use.

  18. Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources

    OpenAIRE

    Olumide A. Odeyemi; Ahmad, Asmat

    2015-01-01

    This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics – Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole...

  19. Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources

    OpenAIRE

    Olumide A. Odeyemi; Asmat Ahmad

    2017-01-01

    This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics – Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole...

  20. Levels of enzymes in leukaemic mice treated withAeromonas L-asparaginase.

    Science.gov (United States)

    Benny, P J; Muraleedhara Kurup, G; Sreejith, K

    1999-07-01

    L-asparaginase isolated in our laboratory fromAeromonas has been found to be antileukaemic. In the present study changes in the levels of serum enzymes in leukaemic mice and under treatment withAeromonas L-asparaginase has been compared. A significant increase in the levels of serum lactate dehydrogenase with tumour growth and a decrease during therapy was observed. A significant decrease in alanine transaminase activity during tumour growth and an increase during treatment was noticed. Increased levels of aspartate transaminase and alkaline phosphatase was observed during enzyme therapy. Total acid phosphatase was found to be increased during tumour growth and decreased considerably during treatment.

  1. Metabolism of proteins and glycoproteins in tumour bearing mice treated with Aeromonas L-asparaginase.

    Science.gov (United States)

    Benny, P J; Muraleedhara, K G; Sreejith, K; Jayashree, G

    1996-12-01

    L-asparaginase, isolated in our laboratory, from Aeromonas had been found to be antileukaemic. In the present study, changes in the levels of proteins and glycoproteins in leukaemic mice and under treatment with Aeromonas L-asparaginase have been compared. Levels of protein bound hexose, fucose and sialic acid which were increased during leukaemia attained normal levels when treated with L-asparaginase. The increased blood urea level declined significantly during enzyme therapy. Effects of L-asparaginase are compared with 'Leunase', a commercially available drug used in the treatment of leukaemia.

  2. Spontaneous aeromonas peritonitis%自发性气单胞菌腹膜炎

    Institute of Scientific and Technical Information of China (English)

    陆天雨

    2011-01-01

    自发性气单胞菌腹膜炎的发生在肝硬化腹水患者中并不罕见.本文就如何诊断及预防自发性气单胞菌腹膜炎,提高患者预后综述如下.%Spontaneous aeromonas peritonitis is a common complication in hepatocirrhosis patients with hepatic ascites.The aim of this review is to discuss how to diagnose and prevent spontaneous Aeromonas peritonitis.

  3. Identification of atypical Aeromonas salmonicida : Inter-laboratory evaluation and harmonization of methods

    DEFF Research Database (Denmark)

    Dalsgaard, Inger; Gudmundsdottir, B.K.; Helgason, S.;

    1998-01-01

    The atypical isolates of Aeromonas salmonicida are becoming increasingly important as the frequency of isolation of bacteria belonging to this group continues to rise. The primary object of this study was to compare and evaluate the results obtained in various laboratories concerning the biochemi......The atypical isolates of Aeromonas salmonicida are becoming increasingly important as the frequency of isolation of bacteria belonging to this group continues to rise. The primary object of this study was to compare and evaluate the results obtained in various laboratories concerning...

  4. Visualizing sensory transmission between dorsal root ganglion and dorsal horn neurons in co-culture with calcium imaging.

    Science.gov (United States)

    Ohshiro, Hiroyuki; Ogawa, Shinji; Shinjo, Katsuhiro

    2007-09-15

    Sensory information is conveyed to the central nervous system by primary afferent neurons within dorsal root ganglia (DRG), which synapse onto neurons of the dorsal horn of the spinal cord. This synaptic connection is central to the processing of both sensory and pain stimuli. Here, we describe a model system to monitor synaptic transmission between DRG neurons and dorsal horn neurons that is compatible with high-throughput screening. This co-culture preparation comprises DRG and dorsal horn neurons and utilizes Ca(2+) imaging with the indicator dye Fura-2 to visualize synaptic transmission. Addition of capsaicin to co-cultures stimulated DRG neurons and led to activation of dorsal horn neurons as well as increased intracellular Ca(2+) concentrations. This effect was dose-dependent and absent when DRG neurons were omitted from the culture. NMDA receptors are a critical component of synapses between DRG and dorsal horn neurons as MK-801, a use-dependent non-competitive antagonist, prevented activation of dorsal horn neurons following capsaicin treatment. This model system allows for rapid and efficient analysis of noxious stimulus-evoked Ca(2+) signal transmission and provides a new approach both for investigating synaptic transmission in the spinal cord and for screening potential analgesic compounds.

  5. Symbiotic Behavior during Co-culturing of Clostridium thermocellum NKP-2 and Thermoanaerobacterium thermosaccharolyticum NOI-1 on Corn Hull

    Directory of Open Access Journals (Sweden)

    Suphavadee Chimtong

    2014-03-01

    Full Text Available The symbiosis of co-culturing between Clostridium thermocellum NKP-2 and Thermoanaerobacterium thermosaccharolyticum NOI-1 is described. An efficient biomass-degrading enriched culture was isolated from soil that contained two different bacterial strains showing homology to C. thermocellum and T. thermosaccharolyticum. The enzymatic system produced from the isolated strains when cultivated individually on corn hulls demonstrated different cellulolytic and xylanolytic enzyme activities. Strain NKP-2 produced cellulose- and xylan-main chain cleaving enzymes such as carboxymethylcellulase (CMCase, avicelase, and xylanase as major enzymes, whereas strain NOI-1 produced primarily short- and side-chain cleaving enzymes such as cellobiohydrolase, β-glucosidase, β-xylosidase, acetyl esterase, and especially α-L-arabinofuranosidase. Enhancement of corn hull utilization, cell growth, and fermentation products (ethanol, butanol, acetic acid, butyric acid, H2, and CO2 was greatly increased during co-culturing compared with individual cultivation of both strains. The symbiotic behavior between both strains was one of mutualism, in which the synergistic degradation of corn hulls by co-action of cellulolytic and xylanolytic enzymes promoted hydrolysis of biomass for growth and fermented products.

  6. Landfill leachate treatment using bacto-algal co-culture: An integrated approach using chemical analyses and toxicological assessment.

    Science.gov (United States)

    Kumari, Moni; Ghosh, Pooja; Thakur, Indu Shekhar

    2016-06-01

    The present study aims to evaluate the feasibility of leachate treatment using a synergistic approach by microalgae and bacteria. Leachate from one of the landfill of Northern India showed the presence of various toxic organic contaminants like naphthalene, benzene, phenol and their derivatives, napthols, pesticides, epoxides, phthalates and halogenated organic compounds. ICP-AES analysis revealed high concentrations of Zn, Cr, Fe, Ni, and Pb beyond the maximum permissible limit of discharge. Bacto-algal co-culture was found to be the most efficient in removal of toxic organic contaminants and heavy metals. Further, detoxification efficiency of bacto-algal treatment was evaluated by Methyl tetrazolium (MTT) assay for cytotoxicity and alkaline comet assay for genotoxicity using hepatoma HepG2 cells. Reduction in toxicity was confirmed by an increase in LC50 by 1.9 fold and reduction in Olive Tail Moment by 40.6 fold after 10 days of treatment. Results of the study indicate bioremediation and detoxification potency of bacto-algal co-culture for leachate treatment.

  7. Survival of Campylobacter jejuni in co-culture with Acanthamoeba castellanii: role of amoeba-mediated depletion of dissolved oxygen.

    Science.gov (United States)

    Bui, Xuan Thanh; Winding, Anne; Qvortrup, Klaus; Wolff, Anders; Bang, Dang Duong; Creuzenet, Carole

    2012-08-01

    Campylobacter jejuni is a major cause of infectious diarrhoea worldwide but relatively little is known about its ecology. In this study, we examined its interactions with Acanthamoeba castellanii, a protozoan suspected to serve as a reservoir for bacterial pathogens. We observed rapid degradation of intracellular C.jejuni in A.castellanii 5 h post gentamicin treatment at 25°C. Conversely, we found that A.castellanii promoted the extracellular growth of C.jejuni in co-cultures at 37°C in aerobic conditions. This growth-promoting effect did not require amoebae - bacteria contact. The growth rates observed with or without contact with amoeba were similar to those observed when C.jejuni was grown in microaerophilic conditions. Preconditioned media prepared with live or dead amoebae cultivated with or without C.jejuni did not promote the growth of C.jejuni in aerobic conditions. Interestingly, the dissolved oxygen levels of co-cultures with or without amoebae - bacteria contact were much lower than those observed with culture media or with C.jejuni alone incubated in aerobic conditions, and were comparable with levels obtained after 24 h of growth of C.jejuni under microaerophilic conditions. Our studies identified the depletion of dissolved oxygen by A.castellanii as the major contributor for the observed amoeba-mediated growth enhancement.

  8. Syntrophic co-culture of aerobic Bacillus and anaerobic Clostridium for bio-fuels and bio-hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Jui-Jen; Ho, Cheng-Yu.; Chen, Wei-En; Huang, Chieh-Chen [Department of Life Sciences, National Chung Hsing University, Taichung (China); Chou, Chia-Hung; Lay, Jiunn-Jyi [Department of Science and Technology, National Kaohsiung First University, Kaohsiung (China)

    2008-10-15

    By using brewery yeast waste and microflora from rice straw compost, an anaerobic semi-solid bio-hydrogen-producing system has been established. For the purpose of industrialization, the major players of both aerobic and anaerobic bacterial strains in the system were isolated and their combination for an effective production of bio-hydrogen and other bio-fuels was examined in this study. The phylogenetic analysis found that four anaerobic isolates (Clostridium beijerinckii L9, Clostridium diolis Z2, Clostridium roseum Z5-1, and C. roseum W8) were highly related with each other and belongs to the cluster I clostridia family, the family that many of solvent-producing strains included. On the other hand, one of the aerobic isolates, the Bacillus thermoamylovorans strain I, shown multiple extracellular enzyme activities including lipase, protease, {alpha}-amylase, pectinase and cellulase, was suggested as a good partner for creating an anaerobic environment and pre-saccharification of substrate for those co-cultured solventogenic clostridial strain. Among these clostridial strains, though C. beijerinckii L9 do not show as many extracellular enzyme activities as Bacillus, but it performs the highest hydrogen-producing ability. The original microflora can be updated to a syntrophic bacterial co-culture system contended only with B. thermoamylovorans I and C. beijerinckii L9. The combination of aerobic Bacillus and anaerobic Clostridium may play the key role for developing the industrialized bio-fuels and bio-hydrogen-producing system from biomass. (author)

  9. A pilot-scale study of biohydrogen production from distillery effluent using defined bacterial co-culture

    Energy Technology Data Exchange (ETDEWEB)

    Vatsala, T.M.; Raj, S. Mohan; Manimaran, A. (Shri AMM Murugappa Chettiar Research Centre, Photosynthesis and Energy Division, Tharamani, Chennai, India, 600)

    2008-10-15

    We evaluated the feasibility of improving the scale of hydrogen (H{sub 2}) production from sugar cane distillery effluent using co-cultures of Citrobacter freundii 01, Enterobacter aerogenes E10 and Rhodopseudomonas palustris P2 at 100 m{sup 3} scale. The culture conditions at 100 ml and 2 L scales were optimized in minimal medium and we observed that the co-culture of the above three strains enhanced H{sub 2} productivity significantly. Results at the 100 m{sup 3} scale revealed a maximum of 21.38 kg of H{sub 2}, corresponding to 10692.6 mol, which was obtained through batch method at 40 h from reducing sugar (3862.3 mol) as glucose. The average yield of H{sub 2} was 2.76 mol mol{sup -1} glucose, and the rate of H{sub 2} production was estimated as 0.53 kg/100 m{sup 3}/h. Our results demonstrate the utility of distillery effluent as a source of clean alternative energy and provide insights into treatment for industrial exploitation. (author)

  10. Performance of non-conventional yeasts in co-culture with brewers' yeast for steering ethanol and aroma production.

    Science.gov (United States)

    van Rijswijck, Irma M H; Wolkers-Rooijackers, Judith C M; Abee, Tjakko; Smid, Eddy J

    2017-08-18

    Increasing interest in new beer types has stimulated the search for approaches to extend the metabolic variation of brewers' yeast. Therefore, we tested two approaches using non-conventional yeast to create a beer with lower ethanol content and a complex aroma bouquet. First, the mono-culture performance was monitored of 49 wild yeast isolates of Saccharomyces cerevisiae (16 strains), Cyberlindnera fabianii (9 strains) and Pichia kudriavzevii (24 strains). Interestingly, both C. fabianii and P. kudriavzevii isolates produced relatively more esters compared with S. cerevisiae isolates, despite their limited fermentation capacity. Next, one representative strain of each species (Sc131, Cf65 and Pk129) was applied as co-culture with brewers' yeast (ratio 1:1). Co-cultures with Cf65 and Pk129 resulted in a beer with lower alcohol content (3.5, 3.8 compared with 4.2% v/v) and relatively more esters. At higher inoculum ratios of Cf65 over brewers' yeast, growth inhibition of brewers' yeast was observed, most likely caused by competition for oxygen between brewers' yeast and Cf65 resulting in a reduced level of ethanol and altered aroma profiles. With this study, we demonstrate the feasibility of using non-conventional yeast species in co-cultivation with traditional brewers' yeast to tailor aroma profiles as well as the final ethanol content of beer. © 2017 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  11. Biological activity of a red-tide alga--A. tamarense under co-cultured condition with bacteria

    Institute of Scientific and Technical Information of China (English)

    SU Jian-qiang; YU Zhi-ming; TIAN Yun; SONG Xiu-xian; HONG Hua-sheng; ZHENG Tian-ling

    2005-01-01

    The relationship between Alexandrium tamarense(Lebour) Balech, one of red-tide alga, and two strains of marine bacteria,Bacillius megaterium(S7 ) and B. halmapulus(S10) isolated from Xiamen Western Sea, was investigated by evaluating the growth state of A. tamarense and the variation of β-glucosidase activity in co-culture system. The results showed the growth and multiplication of the alga were related with the concentration, genus speciality of the bacteria, and growth stage of the alga itself. The growth of A. tamarense was obviously inhibited by S7 and S10 at high concentration. Either inhibition or promotion contributed much more clearly in earlier than in later stage of the growth of the alga. Furthermore, there was a roughly similar variation trend of the activity of extra-cellular enzyme, β-glucosidase, in the water of the separately co-cultured bacteria S7 and S10 with the alga. The β-glucosidase activity(β-GlcA) rapidly increased during the later algal growth accompanying the increase of the lysis of the alga cells. The obvious inhibition of A. tamarense by marine bacteria at high concentration and evident increase of β-GlcA in co-colture system would help us in better understanding the relationship between red-tide alga and bacteria, and also enlightened us the possible use of bacteria in the bio-control of red-tide.

  12. Fabrication of uniform multi-compartment particles using microfludic electrospray technology for cell co-culture studies.

    Science.gov (United States)

    Liu, Zhou; Shum, Ho Cheung

    2013-01-01

    In this work, we demonstrate a robust and reliable approach to fabricate multi-compartment particles for cell co-culture studies. By taking advantage of the laminar flow within our microfluidic nozzle, multiple parallel streams of liquids flow towards the nozzle without significant mixing. Afterwards, the multiple parallel streams merge into a single stream, which is sprayed into air, forming monodisperse droplets under an electric field with a high field strength. The resultant multi-compartment droplets are subsequently cross-linked in a calcium chloride solution to form calcium alginate micro-particles with multiple compartments. Each compartment of the particles can be used for encapsulating different types of cells or biological cell factors. These hydrogel particles with cross-linked alginate chains show similarity in the physical and mechanical environment as the extracellular matrix of biological cells. Thus, the multi-compartment particles provide a promising platform for cell studies and co-culture of different cells. In our study, cells are encapsulated in the multi-compartment particles and the viability of cells is quantified using a fluorescence microscope after the cells are stained for a live/dead assay. The high cell viability after encapsulation indicates the cytocompatibility and feasibility of our technique. Our multi-compartment particles have great potential as a platform for studying cell-cell interactions as well as interactions of cells with extracellular factors.

  13. Interspecies interactions result in enhanced biofilm formation by co-cultures of bacteria isolated from a food processing environment.

    Science.gov (United States)

    Røder, Henriette L; Raghupathi, Prem K; Herschend, Jakob; Brejnrod, Asker; Knøchel, Susanne; Sørensen, Søren J; Burmølle, Mette

    2015-10-01

    Bacterial attachment and biofilm formation can lead to poor hygienic conditions in food processing environments. Furthermore, interactions between different bacteria may induce or promote biofilm formation. In this study, we isolated and identified a total of 687 bacterial strains from seven different locations in a meat processing environment and evaluated their biofilm formation capability. A diverse group of bacteria was isolated and most were classified as poor biofilm producers in a Calgary biofilm device assay. Isolates from two sampling sites, the wall and the meat chopper, were further examined for multispecies biofilm formation. Eight strains from each sampling site were chosen and all possible combinations of four member co-cultures were tested for enhanced biofilm formation at 15 °C and 24 °C. In approximately 20% of the multispecies consortia grown at 15 °C, the biofilm formation was enhanced when comparing to monospecies biofilms. Two specific isolates (one from each location) were found to be present in synergistic combinations with higher frequencies than the remaining isolates tested. This data provides insights into the ability of co-localized isolates to influence co-culture biofilm production with high relevance for food safety and food production facilities.

  14. Astrocyte-neuron co-culture on microchips based on the model of SOD mutation to mimic ALS.

    Science.gov (United States)

    Kunze, Anja; Lengacher, Sylvain; Dirren, Elisabeth; Aebischer, Patrick; Magistretti, Pierre J; Renaud, Philippe

    2013-07-24

    Amyotrophic lateral sclerosis (ALS) is the most common motor neuron disease. ALS is believed to be a non-cell autonomous condition, as other cell types, including astrocytes, have been implicated in disease pathogenesis. Hence, to facilitate the development of therapeutics against ALS, it is crucial to better understand the interactions between astrocytes and neural cells. Furthermore, cell culture assays are needed that mimic the complexity of cell to cell communication at the same time as they provide control over the different microenvironmental parameters. Here, we aim to validate a previously developed microfluidic system for an astrocyte-neuron cell culture platform, in which astrocytes have been genetically modified to overexpress either a human wild-type (WT) or a mutated form of the super oxide dismutase enzyme 1 (SOD1). Cortical neural cells were co-cultured with infected astrocytes and studied for up to two weeks. Using our microfluidic device that prevents direct cell to cell contact, we could evaluate neural cell response in the vicinity of astrocytes. We showed that neuronal cell density was reduced by about 45% when neurons were co-cultured with SOD-mutant astrocytes. Moreover, we demonstrated that SOD-WT overexpressing astrocytes reduced oxidative stress on cortical neurons that were in close metabolic contact. In contrast, cortical neurons in metabolic contact with SOD-mutant astrocytes lost their synapsin protein expression after severe glutamate treatment, an indication of the toxicity potentiating effect of the SOD-mutant enzyme.

  15. Applicability of a Lactobacillus amylovorus strain as co-culture for natural folate bio-enrichment of fermented milk.

    Science.gov (United States)

    Laiño, Jonathan Emiliano; Juarez del Valle, Marianela; Savoy de Giori, Graciela; LeBlanc, Jean Guy Joseph

    2014-11-17

    The ability of 55 strains from different Lactobacillus species to produce folate was investigated. In order to evaluate folic acid productivity, lactobacilli were cultivated in the folate-free culture medium (FACM). Most of the tested strains needed folate for growth. The production and the extent of vitamin accumulation were distinctive features of individual strains. Lactobacillus amylovorus CRL887 was selected for further studies because of its ability to produce significantly higher concentrations of vitamin (81.2 ± 5.4 μg/L). The safety of this newly identified folate producing strain was evaluated through healthy experimental mice. No bacterial translocation was detected in liver and spleen after consumption of CRL887 during 7 days and no undesirable side effects were observed in the animals that received this strain. This strain in co-culture with previously selected folate producing starter cultures (Lactobacillus bulgaricus CRL871, and Streptococcus thermophilus CRL803 and CRL415) yielded a yogurt containing high folate concentrations (263.1 ± 2.4 μg/L); a single portion of which would provide 15% of the recommended dietary allowance. This is the first report where a Lactobacillus amylovorus strain was successfully used as co-culture for natural folate bio-enrichment of fermented milk.

  16. Evaluation of PFOS-mediated neurotoxicity in rat primary neurons and astrocytes cultured separately or in co-culture.

    Science.gov (United States)

    Li, Zhenwei; Liu, Qi; Liu, Chang; Li, Chunna; Li, Yachen; Li, Shuangyue; Liu, Xiaohui; Shao, Jing

    2017-02-01

    Perfluorooctane sulfonate (PFOS) is a potential neurotoxicant reported by epidemiological investigations and experimental studies, while the underlying mechanisms are still unclear. Astrocytes not only support for the construction of neurons, but also conduct neuronal functions through glutamate-glutamine cycle in astrocyte-neuron crosstalk. In the present study, the effect of PFOS exposure on rat primary hippocampal neurons or cortex astrocytes was evaluated. Then the role of the astrocytes in PFOS-induced toxic effect on neurons was explored with astrocyte-neuron co-culture system. Exposure of rat primary hippocampal neurons to PFOS has led to oxidation-antioxidation imbalance, increased apoptosis and abnormal autophagy. The adverse effect of PFOS on rat primary cortex astrocytes manifested in the form of altered extracellular glutamate and glutamine concentrations, decreased glutamine synthase activity, as well as decreased gene expression of glutamine synthase, glutamate transporters and glutamine transporters in the glutamate-glutamine cycle. Especially, the alleviation of PFOS-inhibited neurite outgrowth in neurons could be observed in astrocyte-neuron co-culture system, though the ability of astrocytes in fostering neurite outgrowth was affected by PFOS. These results indicated that both astrocytes and neurons might be the targets of PFOS-induced neurotoxicity, and astrocytes could protect against PFOS-inhibited neurite outgrowth in primary cultured neurons. Our research might render some information in explaining the mechanisms of PFOS-induced neurotoxicity. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Conservation of the biochemical properties of IncA from Chlamydia trachomatis and Chlamydia caviae: oligomerization of IncA mediates interaction between facing membranes.

    Science.gov (United States)

    Delevoye, Cédric; Nilges, Michael; Dautry-Varsat, Alice; Subtil, Agathe

    2004-11-05

    The developmental cycle of Chlamydiaceae occurs in a membrane compartment called an inclusion. IncA is a member of a family of proteins synthesized and secreted onto the inclusion membrane by bacteria. IncA proteins from different species of Chlamydiaceae show little sequence similarity. We report that the biochemical properties of Chlamydia trachomatis and Chlamydia caviae are conserved. Both proteins self-associate to form multimers. When artificially expressed by the host cell, they localize to the endoplasmic reticulum. Strikingly, heterologous expression of IncA in the endoplasmic reticulum completely inhibits concomitant inclusion development. Using truncated forms of IncA from C. caviae, we show that expression of the C-terminal cytoplasmic domain of the protein at the surface of the endoplasmic reticulum is sufficient to disrupt the bacterial developmental cycle. On the other hand, development of a C. trachomatis strain that does not express IncA is not inhibited by artificial IncA expression, showing that the disruptive effect observed with the wild-type strain requires direct interactions between IncA molecules at the inclusion and on the endoplasmic reticulum. Finally, we modeled IncA tetramers in parallel four helix bundles based on the structure of the SNARE complex, a conserved structure involved in membrane fusion in eukaryotic cells. Both C. trachomatis and C. caviae IncA tetramers were highly stable in this model. In conclusion, we show that the property of IncA proteins to assemble into multimeric structures is conserved between chlamydial species, and we propose that these proteins may have co-evolved with the SNARE machinery for a role in membrane fusion.

  18. Dynamics of bone marrow-derived endothelial progenitor cell/mesenchymal stem cell interaction in co-culture and its implications in angiogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Aguirre, A.; Planell, J.A. [Institute for Bioengineering of Catalonia (IBEC), Baldiri Reixac 15-21, 08028 Barcelona (Spain); Dept. of Material Science and Metallurgical Engineering, Technical University of Catalonia (UPC), ETSEIB, Av. Diagonal 647, 08028 Barcelona (Spain); CIBER-BBN, Maria de Luna 11, Ed. CEEI, 50118 Zaragoza (Spain); Engel, E., E-mail: elisabeth.engel@upc.edu [Institute for Bioengineering of Catalonia (IBEC), Baldiri Reixac 15-21, 08028 Barcelona (Spain); Dept. of Material Science and Metallurgical Engineering, Technical University of Catalonia (UPC), ETSEIB, Av. Diagonal 647, 08028 Barcelona (Spain); CIBER-BBN, Maria de Luna 11, Ed. CEEI, 50118 Zaragoza (Spain)

    2010-09-17

    Research highlights: {yields} BM-EPCs and MSCs establish complex, self-organizing structures in co-culture. {yields} Co-culture decreases proliferation by cellular self-regulatory mechanisms. {yields} Co-cultured cells present an activated proangiogenic phenotype. {yields} qRT-PCR and cluster analysis identify new target genes playing important roles. -- Abstract: Tissue engineering aims to regenerate tissues and organs by using cell and biomaterial-based approaches. One of the current challenges in the field is to promote proper vascularization in the implant to prevent cell death and promote host integration. Bone marrow endothelial progenitor cells (BM-EPCs) and mesenchymal stem cells (MSCs) are bone marrow resident stem cells widely employed for proangiogenic applications. In vivo, they are likely to interact frequently both in the bone marrow and at sites of injury. In this study, the physical and biochemical interactions between BM-EPCs and MSCs in an in vitro co-culture system were investigated to further clarify their roles in vascularization. BM-EPC/MSC co-cultures established close cell-cell contacts soon after seeding and self-assembled to form elongated structures at 3 days. Besides direct contact, cells also exhibited vesicle transport phenomena. When co-cultured in Matrigel, tube formation was greatly enhanced even in serum-starved, growth factor free medium. Both MSCs and BM-EPCs contributed to these tubes. However, cell proliferation was greatly reduced in co-culture and morphological differences were observed. Gene expression and cluster analysis for wide panel of angiogenesis-related transcripts demonstrated up-regulation of angiogenic markers but down-regulation of many other cytokines. These data suggest that cross-talk occurs in between BM-EPCs and MSCs through paracrine and direct cell contact mechanisms leading to modulation of the angiogenic response.

  19. Promoted differentiation of cynomolgus monkey ES cells into hepatocyte-like cells by co-culture with mouse fetal liver-derived cells

    Institute of Scientific and Technical Information of China (English)

    Ko Saito; Masahide Yoshikawa; Yukiteru Ouji; Kei Moriya; Mariko Nishiofuku; Shigehiko Ueda; Noriko Hayashi; Shigeaki Ishizaka; Hiroshi Fukui

    2006-01-01

    AIM:To explore whether a co-culture of cynomolgus monkey embryonic stem (cES) cells with embryonic liver cells could promote their differentiation into hepatocytes.METHODS:Mouse fetal liver-derived cells (MFLCs) were prepared as adherent cells from mouse embryos on embryonic d (ED) 14, after which undifferentiated cES cells were co-cultured with MFLCs. The induction of cES cells along a hepatic lineage was examined in MFLCassisted differentiation, spontaneous differentiation,and growth factors (GF) and chemicals-induced differentiations (GF-induced differentiation) using retinoic acid, leukemia inhibitory factor (LIF), FGF2, FGF4,hepatocyte growth factor (HGF), oncostatin M (OSM),and dexamethasone.RESULTS:The mRNA expression of α-fetoprotein,albumin (ALB), α-1-antitrypsin, and hepatocyte nuclear factor 4α was observed earlier in the differentiating cES cells co-cultured with MFLCs, as compared to cES cells undergoing spontaneous differentiation and those subjected to GF-induced differentiation. The expression of cytochrome P450 7a1, a possible marker for embryonic endoderm-derived mature hepatocytes,was only observed in cES cells that had differentiated in a co-culture with MFLCs. Further, the disappearance of Oct3/4, a representative marker of an undifferentiated state, was noted in cells co-cultured with MFLCs, but not in those undergoing spontaneous or GF-induced differentiation. Tmmunocytochemical analysis revealed an increased ratio of ALS-immunopositive cells among cES cells co-cultured with MFLCs, while glycogen storage and urea synthesis were also demonstrated.CONCLUSION:MFLCs showed an ability to induce cES cells to differentiate toward hepatocytes. The co-culture system with MFLCs is a useful method for induction of hepatocyte-like cells from undifferentiated cES cells.

  20. The Manipulation effectivity of cell co-cultures in 5% CO2 incubation system to increase in vitro cattle embryo production

    Directory of Open Access Journals (Sweden)

    Ferry Lismanto Syaiful

    2010-03-01

    Full Text Available The purpose of this research is to determine the effectivity of various cell co-cultures of cattle embryo production by in vitro CO2 5% incubation system and get the best cell co-culture. Cell co-culture which are used in the synthesis is the oviduct cells, isthmus cells, ampulla cells, follicle cells and without cells. Data were analyzed based on completely randomized desiggn. The average growth rate/ cleavage in various cell culture was: the oviduct cell 59.24%, ampulla cell 58.69%, isthmus cell 58.25%, follicle cell 52.24% and without cells 47.76%. The average growth of 8-16 cells embryos to various cell co-culture was: the oviduct cell 46.02%, ampulla cell 45.45%, isthmus cell 45.15%, follicle cell 43.07%, and without cell 38.50%. The mean percentage of morula in various cell co-culture treatment was: the oviduct cell 20.59%, ampulla cell 20.48%, isthmus cell 20.30%, follicle cell 16.96% and without cell 12.58%. The average percentage of embryonic growth (cleavage, 8-16 cells and morula was not significantly different (P > 0.05. The treatment of a variety of cell co-culture increased significantly (P>0.05, blastocysts production, namely: the oviduct cell 3.28%, ampulla cell 3.22%, isthmus cell 3.08%, follicle cell 2.45% and without cell 1.97%. In conclusion, the treatment of various cell co-culture in 5%CO2 incubation system can increace the growth of cattle embryos in vitro.

  1. Direct binding of boar ejaculate and epididymal spermatozoa to porcine epididymal epithelial cells is also needed to maintain sperm survival in in vitro co-culture.

    Science.gov (United States)

    Yeste, Marc; Castillo-Martín, Míriam; Bonet, Sergi; Briz, Maria Dolors

    2012-04-01

    The aim of the present study was to compare the influence of cultured epididymal epithelial cells (EEC) from corpus, caput or cauda, oviductal epithelial cells (OEC) and non-reproductive epithelial cells (LLC-PK1) on function and survival of epididymal and ejaculated spermatozoa, in the latter case to determine whether such influence differed between morphologically normal and abnormal spermatozoa. For this purpose, either spermatozoa were directly co-cultured with EEC from caput, corpus, or cauda, OEC and LLC-PK1 cells (experiment 1) or a membrane-diffusible insert was included in these co-cultures (experiment 2). EEC cultured from the three epididymal regions did not differently affect the sperm parameters. Morphologically normal spermatozoa presented a higher ability to bind EEC, OEC, and LLC-PK1 than abnormal spermatozoa with cytoplasmic droplets or with tail/head malformations. Epididymal spermatozoa were more able to bind EEC during the first 24 h of co-culture, while ejaculated spermatozoa presented a higher capacity to bind OEC between 30 min and 3 h of co-incubation. In all cases, the ability to bind to epithelial cells was higher when they were co-cultured with EEC and OEC than with LLC-PK1. After 2 h of co-culture, the viability of epididymal spermatozoa was better maintained when they bound EEC than when they bound OEC. Conversely, the viability of ejaculated spermatozoa was better maintained when bound OEC than when bound EEC after 24 and 48 h of co-culture. Our work, apart from corroborating the involvement of morphologically normal spermatozoa in the formation of sperm reservoir, highlights the importance of direct contact spermatozoa-EEC in maintaining the sperm survival in in vitro co-culture, and also suggests that a specific binding between EEC and epididymal spermatozoa exists.

  2. Transdifferentiation of mouse adipose-derived stromal cells into acinar cells of the submandibular gland using a co-culture system