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Sample records for cholerae o1 strains

  1. Non-toxigenic environmental Vibrio cholerae O1 strain from Haiti provides evidence of pre-pandemic cholera in Hispaniola

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    Azarian, Taj; Ali, Afsar; Johnson, Judith A.; Jubair, Mohammad; Cella, Eleonora; Ciccozzi, Massimo; Nolan, David J.; Farmerie, William; Rashid, Mohammad H.; Sinha-Ray, Shrestha; Alam, Meer T.; Morris, J. Glenn; Salemi, Marco

    2016-01-01

    Vibrio cholerae is ubiquitous in aquatic environments, with environmental toxigenic V. cholerae O1 strains serving as a source for recurrent cholera epidemics and pandemic disease. However, a number of questions remain about long-term survival and evolution of V. cholerae strains within these aquatic environmental reservoirs. Through monitoring of the Haitian aquatic environment following the 2010 cholera epidemic, we isolated two novel non-toxigenic (ctxA/B-negative) Vibrio cholerae O1. These two isolates underwent whole-genome sequencing and were investigated through comparative genomics and Bayesian coalescent analysis. These isolates cluster in the evolutionary tree with strains responsible for clinical cholera, possessing genomic components of 6th and 7th pandemic lineages, and diverge from “modern” cholera strains around 1548 C.E. [95% HPD: 1532–1555]. Vibrio Pathogenicity Island (VPI)-1 was present; however, SXT/R391-family ICE and VPI-2 were absent. Rugose phenotype conversion and vibriophage resistance evidenced adaption for persistence in aquatic environments. The identification of V. cholerae O1 strains in the Haitian environment, which predate the first reported cholera pandemic in 1817, broadens our understanding of the history of pandemics. It also raises the possibility that these and similar environmental strains could acquire virulence genes from the 2010 Haitian epidemic clone, including the cholera toxin producing CTXϕ. PMID:27786291

  2. Non-toxigenic environmental Vibrio cholerae O1 strain from Haiti provides evidence of pre-pandemic cholera in Hispaniola.

    Science.gov (United States)

    Azarian, Taj; Ali, Afsar; Johnson, Judith A; Jubair, Mohammad; Cella, Eleonora; Ciccozzi, Massimo; Nolan, David J; Farmerie, William; Rashid, Mohammad H; Sinha-Ray, Shrestha; Alam, Meer T; Morris, J Glenn; Salemi, Marco

    2016-10-27

    Vibrio cholerae is ubiquitous in aquatic environments, with environmental toxigenic V. cholerae O1 strains serving as a source for recurrent cholera epidemics and pandemic disease. However, a number of questions remain about long-term survival and evolution of V. cholerae strains within these aquatic environmental reservoirs. Through monitoring of the Haitian aquatic environment following the 2010 cholera epidemic, we isolated two novel non-toxigenic (ctxA/B-negative) Vibrio cholerae O1. These two isolates underwent whole-genome sequencing and were investigated through comparative genomics and Bayesian coalescent analysis. These isolates cluster in the evolutionary tree with strains responsible for clinical cholera, possessing genomic components of 6(th) and 7(th) pandemic lineages, and diverge from "modern" cholera strains around 1548 C.E. [95% HPD: 1532-1555]. Vibrio Pathogenicity Island (VPI)-1 was present; however, SXT/R391-family ICE and VPI-2 were absent. Rugose phenotype conversion and vibriophage resistance evidenced adaption for persistence in aquatic environments. The identification of V. cholerae O1 strains in the Haitian environment, which predate the first reported cholera pandemic in 1817, broadens our understanding of the history of pandemics. It also raises the possibility that these and similar environmental strains could acquire virulence genes from the 2010 Haitian epidemic clone, including the cholera toxin producing CTXϕ.

  3. Cholera outbreaks caused by an altered Vibrio cholerae O1 El Tor biotype strain producing classical cholera toxin B in Vietnam in 2007 to 2008.

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    Nguyen, Binh Minh; Lee, Je Hee; Cuong, Ngo Tuan; Choi, Seon Young; Hien, Nguyen Tran; Anh, Dang Duc; Lee, Hye Ri; Ansaruzzaman, M; Endtz, Hubert P; Chun, Jongsik; Lopez, Anna Lena; Czerkinsky, Cecil; Clemens, John D; Kim, Dong Wook

    2009-05-01

    Vibrio cholerae O1 isolates collected during cholera outbreaks occurring from late 2007 to early 2008 in northern Vietnam were revealed to represent an altered strain containing the RS1 element followed by a CTX prophage harboring El Tor type rstR and classical ctxB on the large chromosome.

  4. Clonal relationship among Vibrio cholerae O1 El Tor strains isolated in Somalia.

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    Scrascia, Maria; Pugliese, Nicola; Maimone, Francesco; Mohamud, Kadigia A; Grimont, Patrick A D; Materu, Sadiki F; Pazzani, Carlo

    2009-03-01

    One hundred and three Vibrio cholerae O1 strains, selected to represent the cholera outbreaks which occurred in Somalia in 1998-1999, were characterized by random amplified polymorphic DNA patterns, ribotyping, and antimicrobial susceptibility. All strains showed a unique amplified DNA pattern and 2 closely related ribotypes (B5a and B8a), among which B5a was the more frequently identified. Ninety-one strains were resistant to ampicillin, chloramphenicol, spectinomycin, streptomycin, sulfamethoxazole, and trimethoprim, conferred, except for spectinomycin, by a conjugative plasmid IncC. These findings indicated that the group of strains active in Somalia in the late 1990s had a clonal origin.

  5. Variations in SXT elements in epidemic Vibrio cholerae O1 El Tor strains in China.

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    Wang, Ruibai; Yu, Dong; Yue, Junjie; Kan, Biao

    2016-03-09

    Vibrio cholerae O1 El Tor biotype strains are responsible for three multiyear epidemics of cholera in China during the seventh ongoing pandemic. The presence of the integrative conjugative element SXT is strongly correlated with resistance to nalidixic acid, tetracycline, and trimethoprim-sulfamethoxazole in these strains. Here, we sequenced the conserved genes of the SXT element, including eex, setR, and int, from 59 V. cholerae O1 El Tor strains and extracted and assembled the intact SXT sequences from the 11 genome sequenced strains. These elements had characteristics distinct from those of previously reported integrative conjugative elements (ICEs). They could be clearly divided into two types based on the clustering of conserved genes and gene structures of the elements, showing their possibly independent derivation and evolution. These two types were present before and after 2005, respectively, demonstrating the type substitution that occurred in 2005. Four to six antibiotic-resistant genes were found on the SXT elements, including genes resistant to tetracycline, trimethoprim-sulfamethoxazole, and multiple drugs. In summary, our findings demonstrated the roles of the SXT element in the emergence of multidrug resistance in epidemic O1 El Tor V. cholerae strains in China.

  6. Whole-Genome Sequencing of Vibrio cholerae O1 El Tor Strains Isolated in Ukraine (2011) and Russia (2014)

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    Smirnova, Nina I.; Agafonova, Elena Y.; Shchelkanova, Elena Y.; Alkhova, Zhanna V.; Kutyrev, Vladimir V.

    2017-01-01

    ABSTRACT Here, we present the draft whole-genome sequence of Vibrio cholerae O1 El Tor strains 76 and M3265/80, isolated in Mariupol, Ukraine, and Moscow, Russia. The presence of various mutations detected in virulence-associated mobile elements indicates high genetic similarity of the strains reported here with new highly virulent variants of the cholera agent V. cholerae. PMID:28232438

  7. Phenotypic diversity of toxigenic Vibrio cholerae O1 E1 Tor strains identified in China

    Institute of Scientific and Technical Information of China (English)

    赵璇

    2014-01-01

    Objective To understand the phenotypic diversity of toxigenic Vibrio cholerae O1 E1 Tor strains isolated from different provinces in China during the last 50 years.Methods Traditional biotyping testings including susceptibility to polymyxin B,sensitivity to groupⅣphage,Voges-Proskauer test and haemolysis of sheep erythrocytes were conducted.Results Data from Biotype-specific phenotype analysis revealed that only 133 isolates carried the typical E1 Tor phenotypes while the other 251

  8. Bactericidal Efficacy of Allium sativum (garlic Against Multidrug Resistant Vibrio cholerae O1 Epidemic Strains

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    Pramod Kumar

    2016-09-01

    Full Text Available In recent years, emerging trend of antibiotic resistance in Vibrio cholerae associated with cholera epidemics is a matter of serious concern for the management of the disease. Indiscriminate use of antibiotics generally results in selection of antibiotic resistant strains. Introduction of newer antibiotics is a challenging task for the researchers as bacteria soon attain resistance. Therefore, identifying natural compounds of medicinal importance for control of cholera would be the best alternative. Garlic (Allium sativum was recognised for many centuries in early Chinese, Egyptian and Indian civilisations as an herbal or traditional medicine. In present study, garlic was selected for screening of antimicrobial efficacy against V. cholerae. A total of 55 V. cholerae strains isolated from various outbreaks/epidemics were subjected to antimicrobial testing as per CLSI, USA 2010 guidelines. Antimicrobial screening of garlic extract was performed against all the multidrug resistant strains of V. cholerae. The garlic extracts showed antibacterial activity against all the V. cholerae strains tested, irrespective of their origin, multidrug resistance and virulence. Antibacterial efficacy of garlic on V. cholerae was also evident from in vivo study on sealed adult mice model. Thus, the Garlic extract harnesses the potential to control infection of multidrug resistant V. cholerae, especially in outbreak like situations in remote and under developed areas where drug supply itself is a challenge

  9. Vibrio cholerae O1 strains are facultative intracellular bacteria, able to survive and multiply symbiotically inside the aquatic free-living amoeba Acanthamoeba castellanii.

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    Abd, Hadi; Saeed, Amir; Weintraub, Andrej; Nair, G Balakrish; Sandström, Gunnar

    2007-04-01

    Vibrio cholerae species are extracellular, waterborne, gram-negative bacteria that are overwhelmed by predators in aquatic environments. The unencapsulated serogroup V. cholerae O1 and encapsulated V. cholerae O139 cause epidemic and pandemic outbreaks of cholera. It has recently been shown that the aquatic and free-living amoeba Acanthamoeba castellanii is not a predator to V. cholerae O139; rather, V. cholerae O139 has shown an intracellular compatibility with this host. The aim of this study was to examine the ability of V. cholerae O1 classical and El Tor strains to grow and survive in A. castellanii. The interaction between A. castellanii and V. cholerae O1 strains was studied by means of amoeba cell counts and viable counts of the bacteria in the absence or presence of amoebae. The viable count of intracellularly growing bacteria was estimated by utilizing gentamicin assay. Confocal microscopy and electron microscopy were used to determine the intracellular localization of V. cholerae in A. castellanii. The results showed that V. cholerae O1 classical and El Tor strains grew and survived intracellularly in the cytoplasm of trophozoites, and that the bacteria were also found in the cysts of A. castellanii. The interaction showed a facultative intracellular behaviour of V. cholerae O1 classical and El Tor strains and a possible role of A. castellanii as an environmental host of V. cholerae species.

  10. Comparative genome analysis of VSP-II and SNPs reveals heterogenic variation in contemporary strains of Vibrio cholerae O1 isolated from cholera patients in Kolkata, India.

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    Imamura, Daisuke; Morita, Masatomo; Sekizuka, Tsuyoshi; Mizuno, Tamaki; Takemura, Taichiro; Yamashiro, Tetsu; Chowdhury, Goutam; Pazhani, Gururaja P; Mukhopadhyay, Asish K; Ramamurthy, Thandavarayan; Miyoshi, Shin-Ichi; Kuroda, Makoto; Shinoda, Sumio; Ohnishi, Makoto

    2017-02-13

    Cholera is an acute diarrheal disease and a major public health problem in many developing countries in Asia, Africa, and Latin America. Since the Bay of Bengal is considered the epicenter for the seventh cholera pandemic, it is important to understand the genetic dynamism of Vibrio cholerae from Kolkata, as a representative of the Bengal region. We analyzed whole genome sequence data of V. cholerae O1 isolated from cholera patients in Kolkata, India, from 2007 to 2014 and identified the heterogeneous genomic region in these strains. In addition, we carried out a phylogenetic analysis based on the whole genome single nucleotide polymorphisms to determine the genetic lineage of strains in Kolkata. This analysis revealed the heterogeneity of the Vibrio seventh pandemic island (VSP)-II in Kolkata strains. The ctxB genotype was also heterogeneous and was highly related to VSP-II types. In addition, phylogenetic analysis revealed the shifts in predominant strains in Kolkata. Two distinct lineages, 1 and 2, were found between 2007 and 2010. However, the proportion changed markedly in 2010 and lineage 2 strains were predominant thereafter. Lineage 2 can be divided into four sublineages, I, II, III and IV. The results of this study indicate that lineages 1 and 2-I were concurrently prevalent between 2007 and 2009, and lineage 2-III observed in 2010, followed by the predominance of lineage 2-IV in 2011 and continued until 2014. Our findings demonstrate that the epidemic of cholera in Kolkata was caused by several distinct strains that have been constantly changing within the genetic lineages of V. cholerae O1 in recent years.

  11. Molecular characterisation of Vibrio cholerae O1 strains carrying an SXT/R391-like element from cholera outbreaks in Kenya: 1994-2007

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    Goddeeris Bruno M

    2009-12-01

    Full Text Available Abstract Background Over the last decade, cholera outbreaks in parts of Kenya have become common. Although a number of recent studies describe the epidemiology of cholera in Kenya, there is paucity of information concerning the diversity and occurrence of mobile genetic elements in Vibrio cholerae strains implicated in these outbreaks. A total of 65 Vibrio cholerae O1 El Tor serotype Inaba isolated between 1994 and 2007 from various outbreaks in Kenya were investigated for mobile genetic elements including integrons, transposons, the integrating conjugative elements (ICEs, conjugative plasmids and for their genotypic relatedness. Results All the strains were haemolytic on 5% sheep blood and positive for the Vibrio cholerae El Tor-specific haemolysin toxin gene (hylA by PCR. They all contained strB, sulII, floR and the dfrA1 genes encoding resistance to streptomycin, sulfamethoxazole, chloramphenicol and trimethoprim respectively. These genes, together with an ICE belonging to the SXT/R391 family were transferable to the rifampicin-resistant E. coli C600 en bloc. All the strains were negative for integron class 1, 2 and 3 and for transposase gene of transposon Tn7 but were positive for integron class 4 and the trpM gene of transposon Tn21. No plasmids were isolated from any of the 65 strains. All the strains were also positive for all V. cholera El Tor pathogenic genes except the NAG- specific heat-stable toxin (st gene. None of the strains were positive for virulence genes associated with the V. cholerae classical biotype. All the strains were positive for El Tor-specific CTXphi bacteriophage rstrR repressor gene (CTXETΦ but negative for the Classical, Calcutta, and the Environmental repressor types. Pulse Field Gel Electrophoresis (PFGE showed that regardless of the year of isolation, all the strains bearing the SXT element were clonally related. Conclusions This study demonstrates that the V. cholerae O1 strains carrying an SXT/R391-like

  12. Comparison of Vibrio cholerae O139 with V. cholerae O1 classical and El Tor biotypes.

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    Calia, K E; Murtagh, M.; Ferraro, M J; Calderwood, S B

    1994-01-01

    Vibrio cholerae O139 is a recently identified non-O1 V. cholerae strain responsible for outbreaks of epidemic cholera in India, Bangladesh, and Thailand in the past 2 years. Other workers have demonstrated the presence of the cholera toxin genetic element in V. cholerae O139, unlike the situation for other non-O1 V. cholerae strains. We sought to compare further this strain with strains of V. cholerae O1, classical and El Tor biotypes, by classic microbiologic methods, Southern blot analysis ...

  13. Vibrio cholerae O1 Strains of Different Ribotypes have Similar hlyA RFLP Patterns but Different Vacuolating Ability

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    Jorge E. Vidal

    2007-01-01

    Full Text Available Extensive cytoplasmic vacuolation on Vero and HeLa cells in vitro by the Vibrio cholerae pore forming toxin HlyA, has been previously reported by our group. Vibrio cholerae O1 and non-O1 pathogenic strains show differences in the potential to induce vacuolation, here we study occurring variations on vacuolating cytotoxic ability, related to changes in the nucleotide sequence of the hlyA-orf. A collection of eight toxigenic strains of V. cholerae O1 El Tor and a non-toxigenic one, all belonging to different ribotypes was tested for their vacuolating ability, and hlyA-orf similarity based on PCR and RFLPs. The strains had extremely different vacuolating capacities, those from the ribotype 2 isolated from the US Gulf Coast, showed the highest vacuolating titer (10240 dil, and the rest of the collection had considerably lower titers ranging among 40 to 360 dilutions. PCR of hlyA-orf, was performed and RFLPs were generated using seven restriction enzymes, this approach later revealed small changes of restriction maps, among the strains. The phenogram constructed from the RFLPs, showed two major branches, one of them included most of the strains, the other separates the only Mexican wild type non-O1 Vibrio cholerae. To test for vacuolating ability out of the Vibrio genetic context, the amplified hlyA-orfs from the collection of strains were cloned in pGEMT- vector system and supernatants from the recombinant E coli DH5-, showed no differences on vacuolating titers, the clones always were low producers. Results from the cloning, together with those from the phenogram indicated that the hlyA gene is mainly conserved and the differences on vacuolating activity are unrelated to minute changes seen in the hlyA-orf. Production of high vacuolating titers on Vibrio strains could be due to transcriptional regulation. Whether the high vacuolating titer would be related to increased virulence, is still to be found.

  14. The study of ctx B and rstR variations of toxigenic Vibrio cholerae O1 E1 Tor strains isolated from 1961 to 2010 in China

    Institute of Scientific and Technical Information of China (English)

    梁未丽

    2014-01-01

    Objective To understand the ctx B and rstR variations of toxigenic Vibrio cholerae(V.cholerae)O1 E1 Tor strains isolated from different provinces in China from1961 to 2010.Methods All 385 toxigenic V.cholerae O1 E1 Tor strains were selected,which were isolated in China between year 1961 and 2010.ctx B gene was amplified by PCR method and sequenced for further analysis.rstR was detected with PCR by using the genotype

  15. Ultrastructural Changes in the Intestine of Suckling Rabbits Infected with Cholerogenic and Non-Cholerogenic nonO1/nonO139 Vibrio cholerae Strains.

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    Monakhova, E V; Fedorenko, G M; Mazrukho, A B; Bardakhch'yan, E A

    2015-09-01

    We performed an electron microscopic study of the small intestine of suckling rabbits infected with cholerogenic and non-cholerogenic strains nonO1/nonO139 Vibrio cholerae. Cholerogenic strain induced mostly hydropic degeneration of the epithelium typical of cholera toxin effect, while non-cholerogenic strain induced the formation of lacunae along the borders of adjacent epithelial cells typical of hemagglutinin/protease effect. In both cases, reduction of microvilli, destruction of intracellular organelles, two types of mitochondrial reaction (condensation and swelling with destruction of cristae), appearance of myelin figures, defects in the capillary walls, and activation of pinocytosis were observed. These data confirm our previous assumption on interchangeability of different pathogenic factors of Vibrio cholerae, including nonO1/nonO139 strains.

  16. Whole genome PCR scanning reveals the syntenic genome structure of toxigenic Vibrio cholerae strains in the O1/O139 population.

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    Bo Pang

    Full Text Available Vibrio cholerae is commonly found in estuarine water systems. Toxigenic O1 and O139 V. cholerae strains have caused cholera epidemics and pandemics, whereas the nontoxigenic strains within these serogroups only occasionally lead to disease. To understand the differences in the genome and clonality between the toxigenic and nontoxigenic strains of V. cholerae serogroups O1 and O139, we employed a whole genome PCR scanning (WGPScanning method, an rrn operon-mediated fragment rearrangement analysis and comparative genomic hybridization (CGH to analyze the genome structure of different strains. WGPScanning in conjunction with CGH revealed that the genomic contents of the toxigenic strains were conservative, except for a few indels located mainly in mobile elements. Minor nucleotide variation in orthologous genes appeared to be the major difference between the toxigenic strains. rrn operon-mediated rearrangements were infrequent in El Tor toxigenic strains tested using I-CeuI digested pulsed-field gel electrophoresis (PFGE analysis and PCR analysis based on flanking sequence of rrn operons. Using these methods, we found that the genomic structures of toxigenic El Tor and O139 strains were syntenic. The nontoxigenic strains exhibited more extensive sequence variations, but toxin coregulated pilus positive (TCP+ strains had a similar structure. TCP+ nontoxigenic strains could be subdivided into multiple lineages according to the TCP type, suggesting the existence of complex intermediates in the evolution of toxigenic strains. The data indicate that toxigenic O1 El Tor and O139 strains were derived from a single lineage of intermediates from complex clones in the environment. The nontoxigenic strains with non-El Tor type TCP may yet evolve into new epidemic clones after attaining toxigenic attributes.

  17. Evidence that a non-O1 Vibrio cholerae produces enterotoxin that is similar but not identical to cholera enterotoxin.

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    Yamamoto, K.; Takeda, Y.; Miwatani, T; Craig, J. P.

    1983-01-01

    Cholera-like enterotoxin produced by a non-O1 strain of Vibrio cholerae, S7 (S7 enterotoxin), isolated from human diarrheal stool, was purified, and its physicochemical, biological, and immunological properties were compared with those of cholera enterotoxin from V. cholerae O1 569B (CT) and an enterotoxin produced by another non-O1 V. cholerae (E8498 enterotoxin) reported previously (Yamamoto et al., Infect. Immun. 39:1128-1135, 1983). The purified S7 enterotoxin had physicochemical properti...

  18. Monitoring water sources for environmental reservoirs of toxigenic Vibrio cholerae O1, Haiti.

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    Alam, Meer T; Weppelmann, Thomas A; Weber, Chad D; Johnson, Judith A; Rashid, Mohammad H; Birch, Catherine S; Brumback, Babette A; Beau de Rochars, Valery E Madsen; Morris, J Glenn; Ali, Afsar

    2014-03-01

    An epidemic of cholera infections was documented in Haiti for the first time in more than 100 years during October 2010. Cases have continued to occur, raising the question of whether the microorganism has established environmental reservoirs in Haiti. We monitored 14 environmental sites near the towns of Gressier and Leogane during April 2012-March 2013. Toxigenic Vibrio cholerae O1 El Tor biotype strains were isolated from 3 (1.7%) of 179 water samples; nontoxigenic O1 V. cholerae was isolated from an additional 3 samples. All samples containing V. cholerae O1 also contained non-O1 V. cholerae. V. cholerae O1 was isolated only when water temperatures were ≥31°C. Our data substantiate the presence of toxigenic V. cholerae O1 in the aquatic environment in Haiti. These isolations may reflect establishment of long-term environmental reservoirs in Haiti, which may complicate eradication of cholera from this coastal country.

  19. Comparative genome analysis of non-toxigenic non-O1 versus toxigenic O1 Vibrio cholerae.

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    Mukherjee, Munmun; Kakarla, Prathusha; Kumar, Sanath; Gonzalez, Esmeralda; Floyd, Jared T; Inupakutika, Madhuri; Devireddy, Amith Reddy; Tirrell, Selena R; Bruns, Merissa; He, Guixin; Lindquist, Ingrid E; Sundararajan, Anitha; Schilkey, Faye D; Mudge, Joann; Varela, Manuel F

    Pathogenic strains of Vibrio cholerae are responsible for endemic and pandemic outbreaks of the disease cholera. The complete toxigenic mechanisms underlying virulence in Vibrio strains are poorly understood. The hypothesis of this work was that virulent versus non-virulent strains of V. cholerae harbor distinctive genomic elements that encode virulence. The purpose of this study was to elucidate genomic differences between the O1 serotypes and non-O1 V. cholerae PS15, a non-toxigenic strain, in order to identify novel genes potentially responsible for virulence. In this study, we compared the whole genome of the non-O1 PS15 strain to the whole genomes of toxigenic serotypes at the phylogenetic level, and found that the PS15 genome was distantly related to those of toxigenic V. cholerae. Thus we focused on a detailed gene comparison between PS15 and the distantly related O1 V. cholerae N16961. Based on sequence alignment we tentatively assigned chromosome numbers 1 and 2 to elements within the genome of non-O1 V. cholerae PS15. Further, we found that PS15 and O1 V. cholerae N16961 shared 98% identity and 766 genes, but of the genes present in N16961 that were missing in the non-O1 V. cholerae PS15 genome, 56 were predicted to encode not only for virulence-related genes (colonization, antimicrobial resistance, and regulation of persister cells) but also genes involved in the metabolic biosynthesis of lipids, nucleosides and sulfur compounds. Additionally, we found 113 genes unique to PS15 that were predicted to encode other properties related to virulence, disease, defense, membrane transport, and DNA metabolism. Here, we identified distinctive and novel genomic elements between O1 and non-O1 V. cholerae genomes as potential virulence factors and, thus, targets for future therapeutics. Modulation of such novel targets may eventually enhance eradication efforts of endemic and pandemic disease cholera in afflicted nations.

  20. Sequences of a co-existing SXT element, a chromosomal integron (CI) and an IncA/C plasmid and their roles in multidrug resistance in a Vibrio cholerae O1 El Tor strain.

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    Wang, Ruibai; Li, Jie; Kan, Biao

    2016-09-01

    The ongoing seventh cholera pandemic is attributed to Vibrio cholerae O1 El Tor biotype strains. Although antibiotic therapy ameliorates symptoms in patients and reduces pathogen transfer to the environment, multidrug resistance remains a major clinical threat. An O1 El Tor strain isolated from a patient in 1998 was intermediate or resistant to 13 antibiotics and could potentially produce extended-spectrum β-lactamase (ESBL), which is very rare in O1 strains. Using genome sequencing, three relevant genetic elements were identified in this strain: a hybrid SXT element (ICEVchCHN1307); a new IncA/C plasmid (pVC1307); and a chromosomal integron. Twenty antibiotic resistance genes were located on them, including blaTEM-1, blaCTX-M-14 and phenotypically silenced tetRA genes. These data elucidate the role of individual genetic components in antibiotic resistance and the accumulation of drug resistance genes in V. cholerae.

  1. Characterization of Vibrio cholerae Strains Isolated from the Nigerian Cholera Outbreak in 2010.

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    Dupke, Susann; Akinsinde, Kehinde A; Grunow, Roland; Iwalokun, Bamidele A; Olukoya, Daniel K; Oluwadun, Afolabi; Velavan, Thirumalaisamy P; Jacob, Daniela

    2016-10-01

    We examined clinical samples from Nigerian patients with acute watery diarrhea for Vibrio cholerae during the 2010 cholera outbreak. A total of 109 suspected isolates were characterized, but only 57 V. cholerae strains could be confirmed using multiplex real-time PCR as well as rpoB sequencing and typed as V. cholerae O:1 Ogawa biotype El Tor. This finding highlighted the need for accurate diagnosis of cholera in epidemic countries to implement life-saving interventions.

  2. Survival of Vibrio cholerae O1 on fomites

    DEFF Research Database (Denmark)

    Farhana, Israt; Hossain, Zenat Zebin; Tulsiani, Suhella Mohan

    2016-01-01

    It is well established that the contamination sources of cholera causing bacteria, Vibrio cholerae, are water and food, but little is known about the transmission role of the fomites (surfaces that can carry pathogens) commonly used in households. In the absence of appropriate nutrients or growth...... conditions on fomites, bacteria have been known to assume a viable but non-culturable (VBNC) state after a given period of time. To investigate whether and when V. cholerae O1 assumes such a state, this study investigated the survival and viable quantification on a range of fomites such as paper, wood, glass......, plastic, cloth and several types of metals under laboratory conditions. The fomites were inoculated with an outbreak strain of V. cholerae and its culturability was examined by drop plate count method at 30 min intervals for up to 6 h. For molecular detection, the viable/dead stain ethidium monoazide (EMA...

  3. 福建省不产毒O1群霍乱弧菌分子特征分析%Molecular characterization of non-toxigenic O1-group Vibrio cholerae strains from Fujian Province

    Institute of Scientific and Technical Information of China (English)

    陈爱平; 李曲文; 倪二茹; 徐海滨; 杨劲松; 郑金凤; 严延生

    2013-01-01

    The majority of Vibrio cholerae isolates from the environment are non-toxigenic strains, which occasionally cause sporadic cholera cases in human. In this study, the distribution of virulence genes including tcpA, rstR, hlyA, zot, ace, toxR and ctxA were investigated in 15 non-toxigenic O1-group V. cholerae isolates from Fujian. It was indicated from PCR amplification that some strains contained minor virulence genes. Among them, eight strains were positive for hly-ET, six strains harbored tcpA-CL, four contained hly-CL, 3 strains were positive for ace gene, 3 were positive for toxR. The rest were negative for other virulence genes. Meanwhile, their genomic diversity were analyzed by pulsed-field gel electrophoresis (PFGE) patterns. Since no dominant PFGE pattern was observed, these strains were classified into 12 PFGE genotypes according to 100% similarity. Based on TENOVER principle, two relatively dominant PFGE group, G1 and G2, were identified. These results suggested that non-toxigenic O1-group V. cholerae harbored considerable virulence genes, no significant difference was observed between isolates from patient and those from environment, and that genomic diversity of these non-toxigenic O1 strains was demonstrated by PFGE profiles.%目的 调查福建省近年从病人和外环境中分离到的不产毒O1群霍乱弧菌的毒力基因分布特征,分析菌株间的遗传相似度.方法 运用PCR扩增技术分别检测15株不产毒O1群霍乱弧菌的毒力相关基因tcpA、rstR、hlyA、zot、ace、toxR、ctxA,并通过脉冲肠凝胶电泳(PFGE)技术进行分子分型.结果 15株不产毒O1群霍乱弧菌各毒力相关基因的阳检数分别为hly-ET(8/15)、tcpA-CL(6/15)、hly-Cl(4/15)、ace(3/15)、toxR(3/15),其余毒力基因均为阴性.按照100%的相似度,PFGE分为12个基因型别,无集中优势的PFGE型别;根据TENOVER原则有两个相对优势的G1、G2 PFGE群.结论 不产毒的O1群霍乱菌株不同程度地携带有相关的

  4. Survival of Vibrio cholerae O1 on fomites.

    Science.gov (United States)

    Farhana, Israt; Hossain, Zenat Zebin; Tulsiani, Suhella Mohan; Jensen, Peter Kjær Mackie; Begum, Anowara

    2016-09-01

    It is well established that the contamination sources of cholera causing bacteria, Vibrio cholerae, are water and food, but little is known about the transmission role of the fomites (surfaces that can carry pathogens) commonly used in households. In the absence of appropriate nutrients or growth conditions on fomites, bacteria have been known to assume a viable but non-culturable (VBNC) state after a given period of time. To investigate whether and when V. cholerae O1 assumes such a state, this study investigated the survival and viable quantification on a range of fomites such as paper, wood, glass, plastic, cloth and several types of metals under laboratory conditions. The fomites were inoculated with an outbreak strain of V. cholerae and its culturability was examined by drop plate count method at 30 min intervals for up to 6 h. For molecular detection, the viable/dead stain ethidium monoazide (EMA) which inhibits amplification of DNA from dead cells was used in combination with real-time polymerase chain reaction (EMA-qPCR) for direct quantitative analyses of viable V. cholerae at 2, 4, 6, 24 h and 7 day time intervals. Results showed that V. cholerae on glass and aluminum surfaces lost culturability within one hour after inoculation but remained culturable on cloth and wood for up to four hours. VBNC V. cholerae on dry fomite surfaces was detected and quantified by EMA-qPCR even 7 days after inoculation. In conclusion, the prolonged survival of V. cholerae on various household fomites may play vital role in cholera transmission and needs to be further investigated.

  5. Novel cholix toxin variants, ADP-ribosylating toxins in Vibrio cholerae non-O1/non-O139 strains, and their pathogenicity.

    Science.gov (United States)

    Awasthi, Sharda Prasad; Asakura, Masahiro; Chowdhury, Nityananda; Neogi, Sucharit Basu; Hinenoya, Atsushi; Golbar, Hossain M; Yamate, Jyoji; Arakawa, Eiji; Tada, Toshiji; Ramamurthy, T; Yamasaki, Shinji

    2013-02-01

    Cholix toxin (ChxA) is a recently discovered exotoxin in Vibrio cholerae which has been characterized as a third member of the eukaryotic elongation factor 2-specific ADP-ribosyltransferase toxins, in addition to exotoxin A of Pseudomonas aeruginosa and diphtheria toxin of Corynebacterium diphtheriae. These toxins consist of three characteristic domains for receptor binding, translocation, and catalysis. However, there is little information about the prevalence of chxA and its genetic variations and pathogenic mechanisms. In this study, we screened the chxA gene in a large number (n = 765) of V. cholerae strains and observed its presence exclusively in non-O1/non-O139 strains (27.0%; 53 of 196) and not in O1 (n = 485) or O139 (n = 84). Sequencing of these 53 chxA genes generated 29 subtypes which were grouped into three clusters designated chxA I, chxA II, and chxA III. chxA I belongs to the prototype, while chxA II and chxA III are newly discovered variants. ChxA II and ChxA III had unique receptor binding and catalytic domains, respectively, in comparison to ChxA I. Recombinant ChxA I (rChxA I) and rChxA II but not rChxA III showed variable cytotoxic effects on different eukaryotic cells. Although rChxA II was more lethal to mice than rChxA I when injected intravenously, no enterotoxicity of any rChxA was observed in a rabbit ileal loop test. Hepatocytes showed coagulation necrosis in rChxA I- or rChxA II-treated mice, seemingly the major target for ChxA. The present study illustrates the potential of ChxA as an important virulence factor in non-O1/non-O139 V. cholerae, which may be associated with extraintestinal infections rather than enterotoxicity.

  6. Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

    Science.gov (United States)

    Sá, L.L.C.; Vale, E.R.V.; Garza, D.R.; Vicente, A.C.P.

    2012-01-01

    Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized. PMID:24031874

  7. Non-O1 Vibrio cholerae in Thailand: homology with cloned cholera toxin genes.

    OpenAIRE

    Hanchalay, S; Seriwatana, J; Echeverria, P.; Holmgren, J.; Tirapat, C.; Moseley, S L; Taylor, D N

    1985-01-01

    We examined 281 non-O1 Vibrio cholerae isolates from Thailand for homology with genes coding for cholera toxin. Five isolates from environmental sources were homologous with the cholera toxin gene probe and produced both the A and B subunits of cholera toxin.

  8. Culture supernatants from V. cholerae O1 ElTor strains isolated from different geographic areas induce cell vacuolation and cytotoxicity Cepas de V. cholerae O1 biotipo ElTor aisladas de diferente origen geográfico inducen vacuolización celular y citotoxicidad

    Directory of Open Access Journals (Sweden)

    Jorge E Vidal

    2009-02-01

    Full Text Available OBJECTIVE: To investigate whether the HlyA-induced vacuolating effect is produced by V. cholerae O1 ElTor strains isolated from different geographic origins, including Mexico. MATERIAL AND METHODS: Supernatant-induced haemolysis, vacuolating activity and cytotoxicity in Vero cells were recorded. PCR, RFLP analysis and molecular cloning were performed. RESULTS: All ElTor strains analyzed induced cellular vacuolation. Ribotype 2 strains isolates from the U.S. gulf coast yielded the highest titer of vacuolating activity. Eight of nine strains were haemolytic, while all strains were PCR positive for the hlyA gene. We cloned the hlyA gene from two ElTor strains, a toxigenic (2514-88, ctxAB+ and a non-toxigenic Mexican strain (CM 91-3, ctxAB-. Supernatant from those recombinant E. coli strains induced haemolysis, cell vacuolation and cytotoxicity. RFLP-PCR analysis revealed similarities in the hlyA gene from all strains tested. CONCLUSION: The HlyA-induced vacuolating effect is a widespread phenotype of epidemic V. cholerae O1 ElTor strains.OBJETIVO: Analizar el efecto vacuolizante de cepas de V. cholerae O1 ElTor aisladas de diferente origen geográfico, incluyendo México. MATERIAL Y MÉTODOS: Se realizaron pruebas de hemolisis, vacuolización y citotoxicidad en células Vero, así como PCR, análisis por RFLP y clonación molecular. RESULTADOS: Todas las cepas indujeron el efecto vacuolizante. Las cepas del ribotipo 2, aisladas de las costas del Golfo en Estados Unidos, presentaron títulos altos de vacuolización. El gen hlyA fue amplificado en las nueve cepas mediante PCR, aunque sólo ocho fueron hemolíticas. Se clonó el gen hlyA de una cepa toxigénica (2514-88, ctxAB+ y de una cepa no toxigénica aislada en México (CM 91-3, ctxAB-. El sobrenadante de las clonas recombinantes indujo hemólisis, efecto vacuolizante y citotoxicidad. El RFLP mostró alta similitud del gen hlyA de las cepas estudiadas. CONCLUSIÓN: El efecto vacuolizante es un

  9. Molecular insights into the evolutionary pathway of Vibrio cholerae O1 atypical El Tor variants.

    Science.gov (United States)

    Kim, Eun Jin; Lee, Dokyung; Moon, Se Hoon; Lee, Chan Hee; Kim, Sang Jun; Lee, Jae Hyun; Kim, Jae Ouk; Song, Manki; Das, Bhabatosh; Clemens, John D; Pape, Jean William; Nair, G Balakrish; Kim, Dong Wook

    2014-09-01

    Pandemic V. cholerae strains in the O1 serogroup have 2 biotypes: classical and El Tor. The classical biotype strains of the sixth pandemic, which encode the classical type cholera toxin (CT), have been replaced by El Tor biotype strains of the seventh pandemic. The prototype El Tor strains that produce biotype-specific cholera toxin are being replaced by atypical El Tor variants that harbor classical cholera toxin. Atypical El Tor strains are categorized into 2 groups, Wave 2 and Wave 3 strains, based on genomic variations and the CTX phage that they harbor. Whole-genome analysis of V. cholerae strains in the seventh cholera pandemic has demonstrated gradual changes in the genome of prototype and atypical El Tor strains, indicating that atypical strains arose from the prototype strains by replacing the CTX phages. We examined the molecular mechanisms that effected the emergence of El Tor strains with classical cholera toxin-carrying phage. We isolated an intermediary V. cholerae strain that carried two different CTX phages that encode El Tor and classical cholera toxin, respectively. We show here that the intermediary strain can be converted into various Wave 2 strains and can act as the source of the novel mosaic CTX phages. These results imply that the Wave 2 and Wave 3 strains may have been generated from such intermediary strains in nature. Prototype El Tor strains can become Wave 3 strains by excision of CTX-1 and re-equipping with the new CTX phages. Our data suggest that inter-chromosomal recombination between 2 types of CTX phages is possible when a host bacterial cell is infected by multiple CTX phages. Our study also provides molecular insights into population changes in V. cholerae in the absence of significant changes to the genome but by replacement of the CTX prophage that they harbor.

  10. Prevalence and molecular characterization of Vibrio cholerae O1, non-O1 and non-O139 in tropical seafood in Cochin, India.

    Science.gov (United States)

    Kumar, Rakesh; Lalitha, Kuttannappilly V

    2013-03-01

    The objective of this study was to determine the prevalence of O1, O139, and non-O1 and non-O139 Vibrio cholerae, which were associated with fresh and raw seafood samples harvested from Cochin, India waters during 2009-2011. Results from V. cholerae-specific biochemical, molecular, and serological assays identified five El Tor V. cholerae O1 Ogawa strains and 377 non-O1, non-O139 V. cholerae strains from 265 seafood samples. V. cholerae O139 strains were not isolated. Polymerase chain reaction assays confirmed the presence of V. cholerae O1 El Tor biotype in seafood. Antibiotic susceptibility analysis revealed that the V. cholerae O1 strains were pansusceptible to 20 test antibiotics, whereas 26%, 40%, 62%, and 84% of the non-O1, non-O139 V. cholerae strains were resistant to cefpodoxime, ticarcillin, augmentin, and colistin, respectively. Detection of virulence and regulatory genes in V. cholerae associated with seafood revealed the presence of virulence and regulatory genes (i.e., ctx, zot, ace, toxR genes) in V. cholerae O1 strains, nevertheless, presence of ace and toxR genes were detected in non-O1, non-O139 in 9.8 and 91% strains, respectively. In conclusion, the presence of pathogenic V. cholerae in seafood harvested from local Cochin waters warrants the introduction of a postharvest seafood monitoring program, which will lead to a greater understanding of the distribution, abundance, and virulence of diverse pathogenic Vibrio populations that inhabit these different coastal regions so that a risk management program can be established.

  11. Peru-15, an improved live attenuated oral vaccine candidate for Vibrio cholerae O1.

    Science.gov (United States)

    Kenner, J R; Coster, T S; Taylor, D N; Trofa, A F; Barrera-Oro, M; Hyman, T; Adams, J M; Beattie, D T; Killeen, K P; Spriggs, D R

    1995-10-01

    Cholera vaccine candidate Peru-15 was derived from a Vibrio cholerae O1 El Tor Inaba strain by deleting the cholera toxin genetic element, introducing the gene encoding cholera toxin B subunit into recA, and screening for nonmotility. In a controlled study, Peru-15 (2 x 10(8) cfu) was administered to 11 volunteers. No vaccinee developed diarrhea, and 10 of 11 had > 4-fold rises in vibriocidal antibody titers. One month later, 5 vaccinees and 5 control volunteers were challenged with wild type V. cholerae O1. Four of 5 controls developed diarrhea (mean, 1.9 L). Two Peru-15 vaccinees developed diarrhea, 1 with volunteer had not developed a significant vibriocidal immune response to vaccination. Peru-15 shows promise as a single-dose, oral cholera vaccine that is safe, immunogenic, and protective.

  12. Multidrug-Resistant Vibrio cholerae O1 was Responsible for a Cholera Outbreak in 2013 in Bagalkot, North Karnataka.

    Science.gov (United States)

    Bhattacharya, Debdutta; Dey, Shuchismita; Roy, Subarna; Parande, Mahantesh V; Telsang, M; Seema, M H; Parande, Aisha V; Mantur, Basappa G

    2015-01-01

    Cholera is a major cause of illness in the developing world. During the monsoon season, small sporadic clusters of cholera cases are reported on an annual basis in Karnataka, India. During the monsoons of 2013, there was a cholera outbreak in Badami, a remote area of Bagalkot district in Karnataka. The multi-drug-resistant Vibrio cholerae O1 serotype Ogawa was found to be responsible for this outbreak. On 5 August 2013, a 30-year-old woman presented with severe dehydration and watery diarrhea at the Aganwadi Health Centre in Badami. A total of 49 suspected cholera cases were reported, with an attack rate of 3.5%. The V. cholerae isolates exhibited resistance to a wide range of drugs, including ampicillin, co-trimoxazole, nitrofurantoin, carbenicillin, and third generation cephalosporins, and showed reduced susceptibility to third generation fluoroquinolones. All of the cephalosporin-resistant V. cholerae strains produced extended-spectrum beta-lactamase. All V. cholerae O1 isolates harbored virulent genes (ctxA, ctxB, tcpA El Tor, Tox S, VPI, ToxT, ToxR, ToxRS, ace, zot, and tcpP) and were found to be genetically similar as determined by randomly amplified polymorphic DNA fingerprinting assay. To the best of our knowledge, this is the first report of a cholera outbreak in the district of Bagalkot. The resistance of V. cholerae to commonly used antimicrobial drugs is becoming a major public health concern in the region as clinicians are left with a limited choice of antibiotics for the treatment of cholera.

  13. Phenotypic diversity of toxigenic Vibrio cholerae O1 El Tor strains identified in China%中国O1群El Tor霍乱弧菌产毒株表型多态性研究

    Institute of Scientific and Technical Information of China (English)

    赵璇; 张力; 李杰; 阚飙; 梁未丽

    2014-01-01

    Objective To understand the phenotypic diversity of toxigenic Vibrio cholerae O1 El Tor strains isolated from different provinces in China during the last 50 years. Methods Traditional biotyping testings including susceptibility to polymyxin B,sensitivity to groupⅣphage, Voges-Proskauer test and haemolysis of sheep erythrocytes were conducted. Results Data from Biotype-specific phenotype analysis revealed that only 133 isolates carryed the typical El Tor phenotypes while the other 251 isolates displayed atypical El Tor phenotypes. Combined with ctxB, rstR genotypes and phenotypic characteristics,64 isolates were identified as typical El Tor biotype,21 were El Tor variants that showing the typical El Tor biotype-specific phenotype but with ctxBclas . 280 isolates were defined as the hybrid groups with traits of both classical and El Tor biotypes that could be further classified into 45 groups,based on the combination of genotypes of ctxB,rstR and phenotypic characteristics. Conclusion Toxigenic Vibrio cholerae O1 El Tor strains that isolated from different provinces in China displayed high phenotypic diversity. The traditional biotype traits could not be used to correctly distinguish the two different biotypes.%目的:了解近50年中国不同地区分离的O1群El Tor型霍乱弧菌产毒株的生物表型特征变化。方法采用多粘菌素B敏感试验、第Ⅳ组霍乱噬菌体裂解试验、VP试验和溶血实验进行表型特征分析。结果生物型表型特征分析表明133株菌具有典型的El Tor生物型表型特征,其余251株菌呈现不典型的El Tor生物型表型特征;综合ctxB、rstR基因型和生物型表型特征分析,385株检测菌株中64株为典型El Tor生物型菌株,21株菌有典型的El Tor生物型表型特征但携带古典型ctxB基因,杂合型特征菌有280株,根据ctxB、rstR和表型特征的不同组合可再分为45个杂合型。结论中国O1群El Tor霍乱弧菌菌株呈现明显的表型

  14. A large cholera outbreak due to a new cholera toxin variant of the Vibrio cholerae O1 El Tor biotype in Orissa, Eastern India.

    Science.gov (United States)

    Kumar, P; Jain, M; Goel, A K; Bhadauria, S; Sharma, S K; Kamboj, D V; Singh, L; Ramamurthy, T; Nair, G B

    2009-02-01

    A total of 32 Vibrio cholerae isolates were collected during a recent large cholera outbreak in Eastern India. Biochemical and serological studies revealed that all of the isolates belonged to serogroup O1, biotype El Tor, serotype Ogawa. Two multiplex PCR assays confirmed the presence of various toxigenic and pathogenic genes - ace, ctxAB, hlyA, ompU, ompW, rfbO1, rtx, tcp, toxR and zot - in all of the isolates. Sequencing of the ctxB gene from the isolates revealed a novel mutation in the gene. Sequencing also confirmed the presence of altered cholera toxin B of the classical biotype in all of the El Tor isolates, suggesting infection of isolates by classical CTXPhi. The molecular diversity of V. cholerae isolates studied by enterobacterial repetitive intergenic consensus sequence PCR, BOX-PCR and randomly amplified polymorphic DNA analysis uniformly showed the clonal relationship among the outbreak V. cholerae O1 isolates. The results of this study suggest that cholera-causing V. cholerae strains are constantly evolving in epidemic areas, highlighting the potential of the emergence of more virulent strains.

  15. Genetic diversities in atypical El Tor strains from Vibrio cholerae O1 serogroup in Fujian Province, China%福建省非典型 O1群埃尔托型霍乱弧菌的研究

    Institute of Scientific and Technical Information of China (English)

    陈爱平; 郑恩惠; 李曲文; 徐海滨; 杨劲松; 王灵岚; 郑金凤; 严延生

    2014-01-01

    The emergence of atypical El Tor strains from V .cholerae in South Asia and Africa has been attributed to several outbreaks in recent decades ,however ,backgrounds of such strains in China remain exclusive .In this study ,PCR am-plification of both El Tor and classical alleles for ctxB ,tcpA ,rstR and hlyA genes was attempted in sixty-nine El Tor isolates from Fujian between 1962 to 2005 ,in addition ,some amplicons were sequence-analyzed .Thus ,the time point of atypical EVC strains in Fujian was determined ,genetic diversities of such strains were investigated .It was revealed that ctxB-Cl ,tcpA-Cl and hlyA genes were detected in O1 serogroup EVC isolates from Fujian since 1962 .Although rstR-Cl gene was solely detected in isolates between 1994 to 2000 .It was indicated by sequence analysis that atypical EVC strains from Fujian possessed a novel T→G mutation at residual 204 of the ctxB gene .Remarkably ,two novel ctxB genotypes (ctxB-10 and ctxB-11) were identified in one strain .The residual 115-C of ctxB in ctxB-11 showed characteristics of ctxB-Cl ,however ,its residual 203-T demonstra-ted characteristics of ctxB-El .This observation implied that it was common in O1 serogroup EVC strains from Fujian hybrid-ized with classical alleles since 1962 ,which would be the earliest time-point for the emergence of atypical El Tor strains hitherto in literature .Emergence of atypical El Tor strains harboring rstR-Cl in Fujian occurred since 1994 .Meanwhile ,novel mutation sites and ctxB genotypes were observed in Fujian isolates ,including diverse combination of ctxB genotypes in one strain and combination of biotype-specific sites in ctxB sequences .In summary ,molecular characterization of O 1 serogroup EVC strains from Fujian was unique and geography-associated .%目的:探索非典型O1群埃尔托型霍乱弧菌在福建省1962-2005年霍乱菌株中的存在及其意义。方法选择1962-2005年代表性霍乱菌株69株(稻叶21、小川48),运用PCR

  16. Genomic epidemiology of Vibrio cholerae O1 associated with floods, Pakistan, 2010.

    Science.gov (United States)

    Shah, Muhammad Ali; Mutreja, Ankur; Thomson, Nicholas; Baker, Stephen; Parkhill, Julian; Dougan, Gordon; Bokhari, Habib; Wren, Brendan W

    2014-01-01

    In August 2010, Pakistan experienced major floods and a subsequent cholera epidemic. To clarify the population dynamics and transmission of Vibrio cholerae in Pakistan, we sequenced the genomes of all V. cholerae O1 El Tor isolates and compared the sequences to a global collection of 146 V. cholerae strains. Within the global phylogeny, all isolates from Pakistan formed 2 new subclades (PSC-1 and PSC-2), lying in the third transmission wave of the seventh-pandemic lineage that could be distinguished by signature deletions and their antimicrobial susceptibilities. Geographically, PSC-1 isolates originated from the coast, whereas PSC-2 isolates originated from inland areas flooded by the Indus River. Single-nucleotide polymorphism accumulation analysis correlated river flow direction with the spread of PSC-2. We found at least 2 sources of cholera in Pakistan during the 2010 epidemic and illustrate the value of a global genomic data bank in contextualizing cholera outbreaks.

  17. Genome assortment, not serogroup, defines Vibrio cholerae pandemic strains

    Energy Technology Data Exchange (ETDEWEB)

    Brettin, Thomas S [Los Alamos National Laboratory; Bruce, David C [Los Alamos National Laboratory; Challacombe, Jean F [Los Alamos National Laboratory; Detter, John C [Los Alamos National Laboratory; Han, Cliff S [Los Alamos National Laboratory; Munik, A C [Los Alamos National Laboratory; Chertkov, Olga [Los Alamos National Laboratory; Meincke, Linda [Los Alamos National Laboratory; Saunders, Elizabeth [Los Alamos National Laboratory; Choi, Seon Y [SEOUL NATL. UNIV.; Haley, Bradd J [U. MARYLAND; Taviani, Elisa [U. MARYLAND; Jeon, Yoon - Seong [INTL. VACCINE INST. SEOUL; Kim, Dong Wook [INTL. VACCINE INST. SEOUL; Lee, Jae - Hak [SEOUL NATL. UNIV.; Walters, Ronald A [PNNL; Hug, Anwar [NATL. INST. CHOLERIC ENTERIC DIS.; Colwell, Rita R [U. MARYLAND

    2009-01-01

    Vibrio cholerae, the causative agent of cholera, is a bacterium autochthonous to the aquatic environment, and a serious public health threat. V. cholerae serogroup O1 is responsible for the previous two cholera pandemics, in which classical and El Tor biotypes were dominant in the 6th and the current 7th pandemics, respectively. Cholera researchers continually face newly emerging and re-emerging pathogenic clones carrying combinations of new serogroups as well as of phenotypic and genotypic properties. These genotype and phenotype changes have hampered control of the disease. Here we compare the complete genome sequences of 23 strains of V. cholerae isolated from a variety of sources and geographical locations over the past 98 years in an effort to elucidate the evolutionary mechanisms governing genetic diversity and genesis of new pathogenic clones. The genome-based phylogeny revealed 12 distinct V. cholerae phyletic lineages, of which one, designated the V. cholerae core genome (CG), comprises both O1 classical and EI Tor biotypes. All 7th pandemic clones share nearly identical gene content, i.e., the same genome backbone. The transition from 6th to 7th pandemic strains is defined here as a 'shift' between pathogenic clones belonging to the same O1 serogroup, but from significantly different phyletic lineages within the CG clade. In contrast, transition among clones during the present 7th pandemic period can be characterized as a 'drift' between clones, differentiated mainly by varying composition of laterally transferred genomic islands, resulting in emergence of variants, exemplified by V.cholerae serogroup O139 and V.cholerae O1 El Tor hybrid clones that produce cholera toxin of classical biotype. Based on the comprehensive comparative genomics presented in this study it is concluded that V. cholerae undergoes extensive genetic recombination via lateral gene transfer, and, therefore, genome assortment, not serogroup, should be used to

  18. Preliminary survey from the antimicrobial resistance of Vibrio cholerae O1, epidemic outbreak 1998

    OpenAIRE

    Ibarra,J.O.; Flores, L. E.; R. H. García; Alvarado,D.E.

    2014-01-01

    Thirty strains of V. cholerae O1 Ogawa and 2 Inaba, isolated during the outbreak of 1998, were studied in relation to thier sensibility in front of 10 antibiotics of clinical use. Nine strains were determined (28.1%) resistant to 1 up to 6 antibiotics. The SIDS-PAGE of the studied strains exhibited similar pattern of bands. Treinta cepas de V. cholerae O1 Ogawa y 2 Inaba, aisladas durante el brote de 1998, fueron estudiados en relación a su sensibilidad frente a 10 antibióticos de uso clín...

  19. Studies on a novel serine protease of a ΔhapAΔprtV Vibrio cholerae O1 strain and its role in hemorrhagic response in the rabbit ileal loop model.

    Directory of Open Access Journals (Sweden)

    Aurelia Syngkon

    Full Text Available BACKGROUND: Two well-characterized proteases secreted by Vibrio cholerae O1 strains are hemagglutinin protease (HAP and V. cholerae protease (PrtV. The hapA and prtV knock out mutant, V. cholerae O1 strain CHA6.8ΔprtV, still retains residual protease activity. We initiated this study to characterize the protease present in CHA6.8ΔprtV strain and study its role in pathogenesis in rabbit ileal loop model (RIL. METHODOLOGY/PRINCIPAL FINDINGS: We partially purified the residual protease secreted by strain CHA6.8ΔprtV from culture supernatant by anion-exchange chromatography. The major protein band in native PAGE was identified by MS peptide mapping and sequence analysis showed homology with a 59-kDa trypsin-like serine protease encoded by VC1649. The protease activity was partially inhibited by 25 mM PMSF and 10 mM EDTA and completely inhibited by EDTA and PMSF together. RIL assay with culture supernatants of strains C6709 (FA ratio 1.1+/-0.3 n = 3, CHA6.8 (FA ratio 1.08+/-0.2 n = 3, CHA6.8ΔprtV (FA ratio 1.02+/-0.2 n = 3 and partially purified serine protease from CHA6.8ΔprtV (FA ratio 1.2+/-0.3 n = 3 induced fluid accumulation and histopathological studies on rabbit ileum showed destruction of the villus structure with hemorrhage in all layers of the mucosa. RIL assay with culture supernatant of CHA6.8ΔprtVΔVC1649 strain (FA ratio 0.11+/-0.005 n = 3 and with protease incubated with PMSF and EDTA (FA ratio 0.3+/-0.05 n = 3 induced a significantly reduced FA ratio with almost complete normal villus structure. CONCLUSION: Our results show the presence of a novel 59-kDa serine protease in a ΔhapAΔprtV V. cholerae O1 strain and its role in hemorrhagic response in RIL model.

  20. OmpU as a biomarker for rapid discrimination between toxigenic and epidemic Vibrio cholerae O1/O139 and non-epidemic Vibrio cholerae in a modified MALDI-TOF MS assay

    NARCIS (Netherlands)

    Paauw, A.; Trip, H.; Niemcewicz, M.; Sellek, R.; Heng, J.M.E.; Mars-Groenendijk, R.H.; Jong, A.L. de; Majchrzykiewicz-Koehorst, J.A.; Olsen, J.S.; Tsivtsivadze, E.

    2014-01-01

    Background Cholera is an acute diarrheal disease caused by Vibrio cholerae. Outbreaks are caused by a genetically homogenous group of strains from serogroup O1 or O139 that are able to produce the cholera toxin. Rapid detection and identification of these epidemic strains is essential for an effecti

  1. Molecular phylogenetic analysis of Vibrio cholerae O1 El Tor strains isolated before, during and after the O 139 outbreak based on the inter-genomic heterogeneity of the 16S-23S rRNA intergenic spacer regions.

    Science.gov (United States)

    Ghatak, Atreyi; Majumdar, Anasuya; Ghosh, Ranajit K

    2005-12-01

    We have cloned, sequenced and analysed all the five classes of the intergenic (16S-23S rRNA) spacer region (ISR) associated with the eight rrn operons (rrna-rrnh) of Vibrio cholerae serogroup O1 El Tor strains isolated before, during and after the O 139 outbreak. ISR classes 'a' and 'g' were found to be invariant, ISR-B (ISRb and ISRe) exhibited very little variation, whereas ISR-C (ISRc, ISRd, and ISRf) and ISRh showed the maximum variation. Phylogenetic analysis conducted with all three ISR classes (ISR-B, ISR-C and ISRh) showed that the pre-O 139 serogroup and post-O 139 serogroup O1 El Tor strains arose out of two independent clones, which was congruent with the observation made by earlier workers suggesting that analyses of ISR-C and ISR-h, instead of all five ISR classes, could be successfully used to study phylogeny in this organism.

  2. Characterization of environmental Vibrio cholerae serogroups O1 and O139 in the Pearl River Estuary, China.

    Science.gov (United States)

    Li, Xiujun; Wang, Duochun; Li, Baisheng; Zhou, Haijian; Liang, Song; Ke, Changwen; Deng, Xiaoling; Kan, Biao; Morris, J Glenn; Cao, Wuchun

    2016-02-01

    Toxigenic isolates of Vibrio cholerae serogroups O1 and O139 from aquatic reservoirs are a key source for recurrent epidemics of cholera in human populations. However, we do not have an optimal understanding of the microbiology of the strains within these reservoirs, particularly outside of the time periods when there are active cholera cases in the surrounding community. The main objective of the present study was to identify and characterize V. cholerae O1 and O139 in the Pearl River Estuary at a time when active disease was not being identified, despite prior occurrence of epidemic cholera in the region. Water samples were collected at 24 sites in the research area at monthly intervals between 2007 and 2010, and screened for the presence of V. cholerae O1 and O139. All isolates were screened for the presence of ctxAB, ompW, toxR, and tcpA genes. Multilocus variable number tandem repeat analysis (MLVA) was used to assess possible relationships among strains. The results show that Vibrio cholerae O1 or O139 was isolated, on average, from 6.7% of the sites screened at each time point. All V. cholerae O1 and O139 isolates were ctxAB negative, and 37% were positive for tcpA. Isolation was most common in the oldest, most urbanized district compared with other districts, and was associated with lower pH. Despite year-to-year variability in isolation rates, there was no evidence of seasonality. MLVA of 27 selected isolates showed evidence of high genetic diversity, with no evidence of clustering by year or geographic location. In this region where cholera has been epidemic in the past, there is evidence of environmental persistence of V. cholerae O1 and O139 strains. However, environmental strains were consistently nontoxigenic, with a high level of genetic diversity; their role as current or future agents of human disease remains uncertain.

  3. Clinical manifestations of non-O1 Vibrio cholerae infections.

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    Yen-Ting Chen

    Full Text Available BACKGROUND: Infections caused by non-O1 Vibrio cholera are uncommon. The aim of our study was to investigate the clinical and microbiological characteristics of patients with non-O1 V. cholera infections. METHODS: The clinical charts of all patients with non-O1 V. cholera infections and who were treated in two hospitals in Taiwan were retrospectively reviewed. RESULTS: From July 2009 to June 2014, a total of 83 patients with non-O1 V. cholera infections were identified based on the databank of the bacteriology laboratories of two hospitals. The overall mean age was 53.3 years, and men comprised 53 (63.9% of the patients. Liver cirrhosis and diabetes mellitus were the two most common underlying diseases, followed by malignancy. The most common type of infection was acute gastroenteritis (n = 45, 54.2%, followed by biliary tract infection (n = 12, 14.5% and primary bacteremia (n = 11, 13.3%. Other types of infection, such as peritonitis (n = 5, 6.0%, skin and soft tissue infection (SSTI (n = 5, 6.0%, urinary tract infection (n = 3, 3.6% and pneumonia (2, 2.4%, were rare. July and June were the most common months of occurrence of V. cholera infections. The overall in-hospital mortality of 83 patients with V. cholera infections was 7.2%, but it was significantly higher for patients with primary bacteremia, hemorrhage bullae, acute kidney injury, acute respiratory failure, or admission to an ICU. Furthermore, multivariate analysis showed that in-hospital mortality was significantly associated with acute respiratory failure (odds ratio, 60.47; 95% CI, 4.79-763.90, P = 0.002. CONCLUSIONS: Non-O1 V. cholera infections can cause protean disease, especially in patients with risk factors and during warm-weather months. The overall mortality of 83 patients with non-O1 V. cholera infections was only 7.2%; however, this value varied among different types of infection.

  4. Toxin(s), Other Than Cholera Toxin, Produced by Environmental Non O1 Non O139 Vibrio cholerae

    Institute of Scientific and Technical Information of China (English)

    Kohinur Begum; Chowdhury R. Ahsan; Mohammad Ansaruzzaman; Dilip K. Dutta; Qazi S.Ahmad; Kaisar A. Talukder

    2006-01-01

    A total of 39 Vibrio cholerae non O1 non O139 strains were isolated from surface waters of different parts of Dhaka City, Bangladesh. All these strains showed lack of ctx or zot gene, as demonstrated by the PCR analysis.Eighteen representative strains were tested for enterotoxin production using a rabbit ileal loop model, of which live cells of 8 strains and culture filtrates of 6 strains produced fluid accumulation in ileal loops. However, none of them produced heat stable toxin (ST), as detected by suckling mouse assay. On the other hand, 15% of isolates produced cytotoxin as detected by the Chinese Hamster Ovary (CHO) cell assay. Fifty times concentrated culture filtrates of the representative strains did not give any precipitin band against the anti-cholera toxin, suggesting the strains produced an enterotoxin, which is antigenically different from known cholera toxin (CT). Eighty percent of the total isolates were found to be positive for heat labile haemolysin detected by tube method, whereas, 39% were found positive by the Christie-Atkins-Munch-Petersen (CAMP) method. However, 87% of the isolates were positive for haemagglutinin/protease and all of the strains were positive for mannose-sensitive-haemagglutinin assay.

  5. Scanning the landscape of genome architecture of non-O1 and non-O139 Vibrio cholerae by whole genome mapping reveals extensive population genetic diversity.

    Science.gov (United States)

    Chapman, Carol; Henry, Matthew; Bishop-Lilly, Kimberly A; Awosika, Joy; Briska, Adam; Ptashkin, Ryan N; Wagner, Trevor; Rajanna, Chythanya; Tsang, Hsinyi; Johnson, Shannon L; Mokashi, Vishwesh P; Chain, Patrick S G; Sozhamannan, Shanmuga

    2015-01-01

    Historically, cholera outbreaks have been linked to V. cholerae O1 serogroup strains or its derivatives of the O37 and O139 serogroups. A genomic study on the 2010 Haiti cholera outbreak strains highlighted the putative role of non O1/non-O139 V. cholerae in causing cholera and the lack of genomic sequences of such strains from around the world. Here we address these gaps by scanning a global collection of V. cholerae strains as a first step towards understanding the population genetic diversity and epidemic potential of non O1/non-O139 strains. Whole Genome Mapping (Optical Mapping) based bar coding produces a high resolution, ordered restriction map, depicting a complete view of the unique chromosomal architecture of an organism. To assess the genomic diversity of non-O1/non-O139 V. cholerae, we applied a Whole Genome Mapping strategy on a well-defined and geographically and temporally diverse strain collection, the Sakazaki serogroup type strains. Whole Genome Map data on 91 of the 206 serogroup type strains support the hypothesis that V. cholerae has an unprecedented genetic and genomic structural diversity. Interestingly, we discovered chromosomal fusions in two unusual strains that possess a single chromosome instead of the two chromosomes usually found in V. cholerae. We also found pervasive chromosomal rearrangements such as duplications and indels in many strains. The majority of Vibrio genome sequences currently in public databases are unfinished draft sequences. The Whole Genome Mapping approach presented here enables rapid screening of large strain collections to capture genomic complexities that would not have been otherwise revealed by unfinished draft genome sequencing and thus aids in assembling and finishing draft sequences of complex genomes. Furthermore, Whole Genome Mapping allows for prediction of novel V. cholerae non-O1/non-O139 strains that may have the potential to cause future cholera outbreaks.

  6. Molecular phylogenetic analysis of Vibrio cholerae O1 El Tor strains isolated before, during and after the O139 outbreak based on the intergenomic heterogeneity of the 16S-23S rRNA intergenic spacer regions

    Indian Academy of Sciences (India)

    Atreyi Ghatak; Anasuya Majumdar; Ranajit K Ghosh

    2005-12-01

    We have cloned, sequenced and analysed all the five classes of the intergenic (16S-23S rRNA) spacer region (ISR) associated with the eight rrn operons (rrna-rrnh) of Vibrio cholerae serogroup O1 El Tor strains isolated before, during and after the O139 outbreak. ISR classes ‘a’ and ‘g’ were found to be invariant, ISR-B (ISRb and ISRe) exhibited very little variation, whereas ISR-C (ISRc, ISRd, and ISRf) and ISRh showed the maximum variation. Phylogenetic analysis conducted with all three ISR classes (ISR-B, ISR-C and ISRh) showed that the pre-O139 serogroup and post-O139 serogroup O1 El Tor strains arose out of two independent clones, which was congruent with the observation made by earlier workers suggesting that analyses of ISR-C and ISR-h, instead of all five ISR classes, could be successfully used to study phylogeny in this organism.

  7. Isolation of New Delhi metallo-β-lactamase 1-producing Vibrio cholerae non-O1, non-O139 strain carrying ctxA, st and hly genes in southern Vietnam.

    Science.gov (United States)

    Diep, Tai The; Nguyen, Nhi Thi Ngoc; Nguyen, Thi Ngoc Cat; An, Huy Khac; Nguyen, Truong Quang; Nguyen, Vu Hoang; Nguyen, Thuong Van; Nguyen, Thu Ngoc Anh; Izumiya, Hidemasa; Ohnishi, Makoto; Yamashiro, Tetsu; Nguyen, Lan Thi Phuong

    2015-05-01

    Vibrio cholerae non-O1, non-O139 (VC_NAG) organisms are universally present in the aquatic environment and regarded as non-pathogenic bacteria. However, considering that they do occasionally induce gastroenteritis, a study of their virulence and antibiotic resistance genes is important. The presence of enteropathogenic genes, including ctxA, VC_NAG-specific heat-stable toxin gene (st), hemolysin (hly), and zona occludens toxin (zot) was determined by PCR in 100 VC_NAG strains isolated in southern Vietnam in 2010-2013 from 94 environmental and six human origins. These 100 VC_NAG strains were also tested phenotypically and genotypically for the presence of the New Delhi metallo-β-lactamase (NDM-1). Of the 100 VC_NAG strains tested, six were positive for ctxA; five from the environment and one of human origin. The st gene was detected in 17 isolates, 15 and two of which were of environmental and human origins, respectively. Gene hly was detected in 19 VC_NAG strains examined, two of which were isolated from humans and 17 from environments. The zot gene was not detected in any of the strains tested. Three VC_NAG strains of environmental origin were confirmed to produce NDM-1 and the blaNDM-1 gene was detected in those strains by PCR. Of note, one of the three NDM-1-producing VC_NAG strains was confirmed to carry ctxA, st and hly genes concurrently. This is the first report of isolation of NDM-1-producing VC_NAG strains in Vietnam.

  8. Rugose atypical Vibrio cholerae O1 El Tor responsible for 2009 cholera outbreak in India.

    Science.gov (United States)

    Chowdhury, Goutam; Bhadra, Rupak K; Bag, Satyabrata; Pazhani, Gururaja P; Das, Bhabatosh; Basu, Pallabi; Nagamani, K; Nandy, Ranjan K; Mukhopadhyay, Asish K; Ramamurthy, Thandavarayan

    2016-10-01

    Vibrio cholerae causes cholera outbreaks in endemic regions where the water quality and sanitation facilities remain poor. Apart from biotype and serotype changes, V. cholerae undergoes phase variation, which results in the generation of two morphologically different variants termed smooth and rugose. In this study, 12 rugose (R-VC) and 6 smooth (S-VC) V. cholerae O1 Ogawa isolates were identified in a cholera outbreak that occurred in Hyderabad, India. Antimicrobial susceptibility results showed that all the isolates were resistant to ampicillin, furazolidone and nalidixic acid. In addition, R-VC isolates were resistant to ciprofloxacin (92 %), streptomycin (92 %), erythromycin (83 %), trimethoprim-sulfamethoxazole (75 %) and tetracycline (75 %). Based on the ctxB gene analysis, all the isolates were identified as El Tor variant with mutation in two positions of ctxB, similar to the classical biotype. The R-VC isolates specifically showed excessive biofilm formation and were comparatively less motile. In addition, the majority of these isolates (~83 %) displayed random mutations in the hapR gene, which encodes haemagglutinin protease regulatory protein. In the PFGE analysis, R-VC and S-VC were placed in distinct clusters but remained clonally related. In the ribotyping analysis, all the R-VC isolates exhibited R-III pattern, which is a prevailing type among the current El Tor isolates. A hapR deletion mutant generated using an S-VC isolate expressed rugose phenotype. To our knowledge, this is the first report on the association of rugose V. cholerae O1 in a large cholera outbreak with extended antimicrobial resistance and random mutations in the haemagglutinin protease regulatory protein encoding gene (hapR).

  9. Distribution of virulence markers in clinical and environmental Vibrio cholerae non-O1/non-O139 strains isolated in Brazil from 1991 to 2000 Distribuição dos marcadores de virulência em cepas clínicas e ambientais de Vibrio cholerae não O1/não O139, isoladas no Brasil no período de 1991 a 2000

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    Grace Nazareth Diogo Theophilo

    2006-04-01

    Full Text Available One hundred seventy nine Vibrio cholerae non-O1/non-O139 strains from clinical and different environmental sources isolated in Brazil from 1991 to 2000 were serogrouped and screened for the presence of four different virulence factors. The Random Amplification of Polymorphic DNA (RAPD technique was used to evaluate the genetic relatedness among strains. Fifty-four different serogroups were identified and V. cholerae O26 was the most common (7.8%. PCR analysis for three genes (ctxA, zot, ace located of the CTX genetic element and one gene (tcpA located on the VPI pathogenicity island showed that 27 strains harbored one or more of these genes. Eight (4.5% strains possessed the complete set of CTX element genes and all but one of these belonged to the O26 serogroup suggesting that V. cholerae O26 has the potential to be an epidemic strain. The RAPD profiles revealed a wide variability among strains and no genetic correlation was observed.Cento e setenta e nove amostras de V. cholerae não O1/não O139, isoladas de casos clínicos (139 e de meio ambiente (40, no período de 1991 a 2000 no Brasil, foram caracterizadas antigenicamente pelo National Institute of Health (Japão e investigadas quanto ao seu potencial genético de virulência, representado pelos genes ctxA, zot, ace e tcpA. As análises fenotípicas revelaram extraordinária diversidade antigênica, com a ocorrência de 54 diferentes sorogrupos, com prevalência para O26 (7,8%. A técnica de PCR, empregada na detecção dos genes localizados no elemento genético CTX (ctxA, zot, ace e na Ilha de Patogenicidade de Vibrio-VPI (tcpA, possibilitou a identificação de 27 cepas contendo qualquer um desses genes. O gene ctxA (codificador da sub-unidade A de CT, só foi evidenciado no sorogrupo O26, sendo também o único capaz de se apresentar com o cassete de virulência de forma intacta. Com base nos resultados obtidos deste estudo preliminar, admite-se a hipótese da potencialidade destas

  10. Population structure and evolution of non-O1/non-O139 Vibrio cholerae by multilocus sequence typing.

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    Sophie Octavia

    Full Text Available Pathogenic non-O1/non-O139 Vibrio cholerae strains can cause sporadic outbreaks of cholera worldwide. In this study, multilocus sequence typing (MLST of seven housekeeping genes was applied to 55 non-O1/non-O139 isolates from clinical and environmental sources. Data from five published O1 isolates and 17 genomes were also included, giving a total of 77 isolates available for analysis. There were 66 sequence types (STs, with the majority being unique, and only three clonal complexes. The V. cholerae strains can be divided into four subpopulations with evidence of recombination among the subpopulations. Subpopulations I and III contained predominantly clinical strains. PCR screening for virulence factors including Vibrio pathogenicity island (VPI, cholera toxin prophage (CTXΦ, type III secretion system (T3SS, and enterotoxin genes (rtxA and sto/stn showed that combinations of these factors were present in the clinical isolates with 85.7% having rtxA, 51.4% T3SS, 31.4% VPI, 31.4% sto/stn (NAG-ST and 11.4% CTXΦ. These factors were also present in environmental isolates but at a lower frequency. Five strains previously mis-identified as V. cholerae serogroups O114 to O117 were also analysed and formed a separate population with V. mimicus. The MLST scheme developed in this study provides a framework to identify sporadic cholera isolates by genetic identity.

  11. The mechanism of applying lysogenicity in phage-biotyping scheme for subtyping O1 E1 Tor Vibrio cholerae strains%O1群E1Tor型霍乱弧菌溶原性测定的机理研究

    Institute of Scientific and Technical Information of China (English)

    申小娜; 张京云; 付秀萍; 李杰; 梁未丽; 阚飙

    2016-01-01

    Objective To determine the principle of applying lysogenicity in Phage-Biotyping Scheme developed for the subtyping of O1 E1 Tor Vibrio cholerae strains.Methods 118 V.cholerae strains including 76 E1 Tor strains,8 classical strains and 34 serogroup O139 were selected to analyze the lysogenicity and sensitivity to lysogenic phage 919TP as described in the Manuals of cholera prevention and control,the genes of this phage were also determined among the genome sequences of these strains and the phages produced by them.Results All O1 E1Tor Vibrio cholerae 19 strains that produced positive results in lysogenicity,had the lysogenic K139 phage in genome and could both resist to lysogenic phage 919TP and release the K139 family phage.All the O1 E1Tor Vibrio cholerae 22 strains that produced positive results in sensitivity to the lysogenic phage,had no K139 family phage genes and got negative results in lysogenicity.However,the phages of this family were not released from 6 classical strains with positive lysogenicity result.Five serogroup O139 strains were detected releasing temperate phages K139 without the sensitivity to phage 919TP.Conclusions Applying the lysogenicity in Phage-Biotyping Scheme for subtyping O1 E1 Tor Vibrio cholerae strains is based on the ability to produce lysogenic bacteriophage K139.The index of "sensitivity to the lysogenic phage" was also associated with this ability.%目的 确定我国O1群El Tor生物型霍乱弧菌“噬菌体-生物分型”方案中“溶原性测定”中检测噬菌体的种类,研究溶原性测定的原理.方法 选择O1群El Tor型霍乱弧菌76株、O1群古典株8株、O139群菌株34株,利用溶原性测定方法检测菌株溶原性,检测菌株自发产生的溶原性噬菌体种类,检测阳性和阴性菌株染色体中该前噬菌体基因组,并检测这些菌株对溶原性噬菌体919TP(弧菌噬菌体K139家族)的敏感性.结果 O1群El Tor型菌株溶原性测定为阳性的19株菌,其释

  12. Growth of Vibrio cholerae O1 Ogawa Eltor in freshwater.

    Science.gov (United States)

    Vital, Marius; Füchslin, Hans Peter; Hammes, Frederik; Egli, Thomas

    2007-07-01

    Growth of Vibrio cholerae O1 Ogawa Eltor was studied with a growth assay in which autoclaved and filtered (0.22 microm) freshwater was inoculated at low cell density (5 x 10(3) cells ml(-1)) and proliferation was followed with flow cytometry. Against the common view, V. cholerae was able to grow extensively in different kinds of freshwater. The bacterium multiplied in river water, lake water and effluent of a wastewater treatment plant up to a cell density of 1.55 x 10(6) cells ml(-1). In these samples, apparent assimilable organic carbon (AOC(app)) concentrations ranged from 52 up to 800 microg l(-1) and the results demonstrate a positive trend between the AOC(app) concentration and final cell concentration, suggesting that AOC was a key parameter governing growth of V. cholerae. No growth was observed in waters (tap and bottled drinking water) containing less than approximately 60 microg AOC(app) l(-1). When pure cultures of V. cholerae were grown on identical lake water at different temperatures (20, 25 and 30 degrees C) the maximum specific growth rates (micromax) achieved were 0.22 h(-1), 0.32 h(-1) and 0.45 h(-1), respectively. In addition, growth was characterized in lake water samples amended with different concentrations of NaCl. The highest micromax of V. cholerae was recorded at moderate salinity levels (5 g NaCl l(-1), micromax=0.84 h(-1)), whereas at 30 g NaCl l(-1) (micromax=0.30 h(-1)) or 0 g NaCl l(-1) (micromax)=0.40 h(-1)) specific growth rates were significantly reduced. In the water tested here, micro(max) of V. cholerae was always around 50 % of that exhibited by a freshwater community of indigenous bacteria enriched from the water sampling site. Direct batch competition experiments between V. cholerae and the lake water bacterial community were performed at different temperatures in which V. cholerae was enumerated in the total community using fluorescent-surface antibodies. In all cases V. cholerae was able to grow and constituted around 10

  13. Antibody-secreting cell responses after Vibrio cholerae O1 infection and oral cholera vaccination in adults in Bangladesh.

    Science.gov (United States)

    Rahman, Atiqur; Rashu, Rasheduzzaman; Bhuiyan, Taufiqur Rahman; Chowdhury, Fahima; Khan, Ashraful Islam; Islam, Kamrul; LaRocque, Regina C; Ryan, Edward T; Wrammert, Jens; Calderwood, Stephen B; Qadri, Firdausi; Harris, Jason B

    2013-10-01

    Infection with Vibrio cholerae and oral cholera vaccines (OCVs) induce transient circulating plasmablast responses that peak within approximately 7 days after infection or vaccination. We previously demonstrated that plasmablast responses strongly correlate with subsequent levels of V. cholerae-specific duodenal antibodies up to 6 months after V. cholerae infection. Hence, plasmablast responses provide an early window into the immunologic memory at the mucosal surface. In this study, we characterized plasmablast responses following V. cholerae infection using a flow cytometrically defined population and compared V. cholerae-specific responses in adult patients with V. cholerae O1 infection and vaccinees who received the OCV Dukoral (Crucell Vaccines Canada). Among flow cytometrically sorted populations of gut-homing plasmablasts, almost 50% of the cells recognized either cholera toxin B subunit (CtxB) or V. cholerae O1 lipopolysaccharide (LPS). Using a traditional enzyme-linked immunosorbent spot assay (ELISPOT), we found that infection with V. cholerae O1 and OCVs induce similar responses to the protein antigen CtxB, but responses to LPS were diminished after OCV compared to those after natural V. cholerae infection. A second dose of OCV on day 14 failed to boost circulating V. cholerae-specific plasmablast responses in Bangladeshi adults. Our results differ from those in studies from areas where cholera is not endemic, in which a second vaccination on day 14 significantly boosts plasmablast responses. Given these results, it is likely that the optimal boosting strategies for OCVs differ significantly between areas where V. cholerae infection is endemic and those where it is not.

  14. Nontoxigenic Vibrio cholerae non-O1/O139 isolate from a case of human gastroenteritis in the U.S. Gulf Coast.

    Science.gov (United States)

    Hasan, Nur A; Rezayat, Talayeh; Blatz, Peter J; Choi, Seon Young; Griffitt, Kimberly J; Rashed, Shah M; Huq, Anwar; Conger, Nicholas G; Colwell, Rita R; Grimes, D Jay

    2015-01-01

    An occurrence of Vibrio cholerae non-O1/O139 gastroenteritis in the U.S. Gulf Coast is reported here. Genomic analysis revealed that the isolate lacked known virulence factors associated with the clinical outcome of a V. cholerae infection but did contain putative genomic islands and other accessory virulence factors. Many of these factors are widespread among environmental strains of V. cholerae, suggesting that there might be additional virulence factors in non-O1/O139 V. cholerae yet to be determined. Phylogenetic analysis revealed that the isolate belonged to a phyletic lineage of environmental V. cholerae isolates associated with sporadic cases of gastroenteritis in the Western Hemisphere, suggesting a need to monitor non-O1/O139 V. cholerae in the interest of public health.

  15. Proteome analysis of the total protein in epidemic and non-epidemic Vibrio cholerae O1 Serotype Ogawa strains by 2-DE%霍乱弧菌O1群稻叶型流行株和非流行株的蛋白质谱特征分析

    Institute of Scientific and Technical Information of China (English)

    林静佳; 李军涛; 邓志爱; 陈守义

    2012-01-01

    Objective To study the differences on proteins between non-epidemic strain and epidemic strain of Vibrw cholerae(V.cholerae) by two dimensional electrophoresis(2-DE) and TOF-TOF-MS. Methods Total protein was extracted from V. cholerae, and separated by 2-DE under immobilized pH gradients(IPG),and then stained with Coomassie Brilliant Blue, and analyzed by ImageMaster 2D Elite 5.0. Finally the different protein spots were identified by TOF-TOF-MS. Results High repetitive 2-DE maps were obtained.2-DE and image analysis revealed (1.081±16) protein spots,and pI value was mainly localized between 4.0 and 7.2.Matching spots were (1.057±28) in two repeats electrophoregrams, with a matching ratio of 97.85%. Conclusion The different protein spots were successfully identified with high quality and sharpness separation by 2-DE and TOF-TOF-MS, and serve as a valuable resource for proteomics research of V.cholerae.%目的 探讨霍乱弧菌O1群稻叶型流行株和非流行株的蛋白表达的差异.方法 利用裂解液处理霍乱弧菌,提取全菌蛋白;采用pH梯度等电聚焦对菌蛋白进行双向电泳;考马斯亮蓝染色后获得的双向电泳图谱,并利用ImageMaster 2D Elite 5.0图象分析软件进行分析,找出差异蛋白;在此基础上,胰蛋白酶消化这些特殊差异蛋白,并进行质谱( MALDI-TOF-MS)分析.结果 获得了(1081±16)个蛋白斑点,蛋白主要集中在pI 4.00~7.20之间,重复胶的匹配点数为(1057±28),匹配率为97,85%;发现了有明显差异的21个蛋白点,并进行了质谱鉴定.结论 获得了霍乱弧菌O1群稻叶型流行株和非流行株的差异表达蛋白.

  16. Cholera

    OpenAIRE

    Harris, Jason B.; LaRocque, Regina C; Qadri, Firdausi; Edward T. Ryan; Calderwood, Stephen B.

    2012-01-01

    Cholera is an acute, secretory diarrhea caused by infection with Vibrio cholerae of the O1 and O139 serogroups. Cholera is endemic in over 50 countries and also causes large epidemics. Since 1817, seven cholera pandemics have spread from Asia to much of the world. The 7th pandemic began in 1961 and affects 3–5 million people each year, killing 120,000. Although mild cholera may be indistinguishable from other diarrheal illnesses, the presentation of severe cholera is distinct, with dramatic d...

  17. Antibody-Conjugated Rubpy Dye-Doped Silica Nanoparticles as Signal Amplification for Microscopic Detection of Vibrio cholerae O1

    Directory of Open Access Journals (Sweden)

    Nualrahong Thepwiwatjit

    2013-01-01

    Full Text Available This study demonstrated the potential application of antibody-conjugated Rubpy dye-doped silica nanoparticles for immunofluorescence microscopic detection of Vibrio cholerae O1. The particle synthesis of 20X of the original ratio was accomplished yielding spherical nanoparticles with an average size of 45±3 nm. The nanoparticles were carboxyl functionalized and then conjugated with either monoclonal antibody or polyclonal antibody against V. cholerae O1. The antibody-conjugated nanoparticles were tested with two target bacteria and three challenge strains. The result showed that monoclonal antibody-conjugated Rubpy dye-doped silica nanoparticles could be effectively used as signal amplification to detect V. cholerae O1 under a fluorescence microscope. Their extremely strong fluorescence signal also enables the detection of a single cell bacterium.

  18. Multidrug-resistant Vibrio cholerae O1 in Belgaum, south India.

    Science.gov (United States)

    Roy, Subarna; Parande, M V; Mantur, B G; Bhat, S; Shinde, R; Parande, A M; Meti, Rajanish S; Chandrasekhar, M R; Kholkute, S D; Saini, A; Joshi, M; Gaonkar, A

    2012-11-01

    An outbreak of acute diarrhoea occurred in the Belgundi area (population 3896) of Belgaum Taluka (population 815 581) in Karnataka, South India, in June 2010. An estimated 16.22 % of people were affected and 0.16 % deaths were reported. Vibrio cholerae O1 El Tor was isolated from 18 of the 147 stool samples cultured. Seven out of eight drinking water samples collected from different sources were found to be grossly contaminated with faecal coliforms. All isolates were multidrug resistant, with some showing resistance to quinolones, gentamicin and cephalosporins in addition to co-trimoxazole and tetracycline, the drugs that were being used by the state health authorities for empirical treatment. Two serotypes and at least eight genotypes of V. cholerae were observed among the isolates. Cholera was confirmed as one, if not the only, cause of the outbreak, which, to our belief, is the first report of cholera from this region. It might have occurred due to a 'flare up' in the number of endemic strains triggered by shortage of portable water, onset of monsoon rains and breakdown of sanitation systems, rather than being a de novo outbreak arising out of new exogenous infectious sources. A change in the empirical treatment, coupled with chlorination, improvement in sanitation measures and extensive Information Education Communication activities, resulted in decline of the outbreak and prevention of further deaths.

  19. [Surveys on the contamination of marine fish with non-O1 Vibrio cholerae and Vibrio mimicus and food poisoning cases by these organisms].

    Science.gov (United States)

    Kodama, H; Hayashi, M; Gyobu, Y

    1991-02-01

    The present paper describes the relationship between the contamination with non-O1 Vibrio cholerae and Vibrio mimicus of marine fish, with special reference to the seasonal variation and the concentration of contamination, and the actual cases of domestic food poisoning by these organisms. A 10 year survey revealed that non-O1 Vibrio cholerae (non-O1 V. cholerae) strains were frequently isolated from fish during the summer season with some variations from one year to another, and isolates from fish showed similar biological properties to those of isolates from diarrhea cases of over-sea travellers. Experimentally enteropathogenic strains were included among these isolates. Vibrio mimicus (V. mimicus) strains were also isolated from fish, the frequency being not so high as in the case of non-O1 V. cholerae Strains of serovar O-41 which was most predominant among strains from diarrhea cases were also detected among the isolates from fish. The viable cell counts, however, were very small with regard to both non-O1 V. cholerae and V. mimicus From these observations, factors causing food poisoning by non-O1 V. cholerae or V. mimicus seemed to be essentially similar to those by Vibrio parahaemolyticus (V. parahaemolyticus); that is, the food poisoning by non-O1 V. cholerae or V. mimicus is apt to occur in the summer season and is caused by the consumption of raw fish, although the frequency might be significantly low in comparison to that of V. parahaemolyticus. The actual cases of the domestic food poisoning by non-O1 V. cholerae or V. mimicus were retrospectively surveyed by the literature.(ABSTRACT TRUNCATED AT 250 WORDS)

  20. A Natural Vaccine Candidate Strain Against Cholera

    Institute of Scientific and Technical Information of China (English)

    LIUYAN-QING; QIGUO-MING; 等

    1995-01-01

    El Tor Vibrio cholerae(EVC)strains may be classified into two kinds-epidemigenic(EEVC)strains and non-epidemigenic(NEEVC)strains-based on a phage-biotyping system.A large number of EEVC strains have been screened for toxigenic and putative colonization attributes.One such naturally occurring strain(designated IEM101)has been found which is devoid of genes encoding cholera toxin(CT),accessory cholera enterotoxin(ACE),zonula occludens toxin(ZOT),but possesses RS1 sequences and toixn-coregulated pilus A gene(tcpA)although tcpA is poorly expressed.It expresses type B pili but does not posses type C pili.It is an El Tor Ogawa strain and does not cause fluid accumulation in rabbit ileal loop tests.Active immunization of rabbits with strain IEM101 elicited good protection against challenge with virulent strains of V.cholerae Ol.Oral administration cased no side effects in 15 human volunteers.colonized the gut for four to ten days and elicited good immune responses.

  1. Genomic and phenotypic characterization of Vibrio cholerae non-O1 isolates from a US Gulf Coast cholera outbreak.

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    Bradd J Haley

    Full Text Available Between November 2010, and May 2011, eleven cases of cholera, unrelated to a concurrent outbreak on the island of Hispaniola, were recorded, and the causative agent, Vibrio cholerae serogroup O75, was traced to oysters harvested from Apalachicola Bay, Florida. From the 11 diagnosed cases, eight isolates of V. cholerae were isolated and their genomes were sequenced. Genomic analysis demonstrated the presence of a suite of mobile elements previously shown to be involved in the disease process of cholera (ctxAB, VPI-1 and -2, and a VSP-II like variant and a phylogenomic analysis showed the isolates to be sister taxa to toxigenic V. cholerae V51 serogroup O141, a clinical strain isolated 23 years earlier. Toxigenic V. cholerae O75 has been repeatedly isolated from clinical cases in the southeastern United States and toxigenic V. cholerae O141 isolates have been isolated globally from clinical cases over several decades. Comparative genomics, phenotypic analyses, and a Caenorhabditis elegans model of infection for the isolates were conducted. This analysis coupled with isolation data of V. cholerae O75 and O141 suggests these strains may represent an underappreciated clade of cholera-causing strains responsible for significant disease burden globally.

  2. Trends in the genomic epidemiology of Vibrio cholerae O1 isolated worldwide since 1961.

    Science.gov (United States)

    Jaiswal, Abhishek; Sarkar, Sounak; Das, Parijat; Nandy, Suman; Koley, Hemanta; Sarkar, Banwarilal

    2015-10-01

    Here we describe the international scenario of Vibrio cholerae with a comparative analysis of different aspects of typing. Representative V. cholerae strains (n=108) associated with endemic cholera regions from 29 states of India and worldwide were subjected to microbiological, molecular and phylogenetic study. All of the strains were V. cholerae serogroup O1 biotype El Tor and were typed according to both the new phage (NP) type and Basu & Mukherjee (BM) typing schemes. The predominant phage type was T-27 (NP)/T-4 (BM) (65.7%; n=71), followed by phage type T-27 (NP)/T-2 (BM) (14.8%; n=16), T-26 (NP)/T4 (BM) (12.0%; n=13), T-13 (NP)/T-4 (BM) (2.8%; n=3), T-20 (NP)/T-4 (BM) (1.9%; n=2), T-3 (NP)/T-4 (BM) (0.9%; n=1), T-23 (NP)/T-4 (BM) (0.9%; n=1) and T-24 (NP)/T-2 (BM) (0.9%; n=1). Mismatch amplification mutation assay PCR (MAMA-PCR) findings showed the dominance of ctxB El Tor genotype (77.1%; 54/70) from 1961-1991, whilst the next two epochs showed the supremacy of ctxB classical genotype. Multidrug-resistant strains showed resistance to erythromycin, streptomycin, trimethoprim/sulfamethoxazole, norfloxacin and ampicillin. The regional resistance of epidemic clones in India draws a layout of the rapid dissemination of resistance in the past 30 years and the necessity of proper treatment to protect populations at risk.

  3. Whole-genome sequence comparisons reveal the evolution of Vibrio cholerae O1.

    Science.gov (United States)

    Kim, Eun Jin; Lee, Chan Hee; Nair, G Balakrish; Kim, Dong Wook

    2015-08-01

    The analysis of the whole-genome sequences of Vibrio cholerae strains from previous and current cholera pandemics has demonstrated that genomic changes and alterations in phage CTX (particularly in the gene encoding the B subunit of cholera toxin) were major features in the evolution of V. cholerae. Recent studies have revealed the genetic mechanisms in these bacteria by which new variants of V. cholerae are generated from type-specific strains; these mechanisms suggest that certain strains are selected by environmental or human factors over time. By understanding the mechanisms and driving forces of historical and current changes in the V. cholerae population, it would be possible to predict the direction of such changes and the evolution of new variants; this has implications for the battle against cholera.

  4. The Aquatic Environment as a Reservoir of Vibrio cholerae O1 in Hydrographic Basins of the State of Pernambuco, Brazil

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    Carina Lucena Mendes-Marques

    2013-01-01

    Full Text Available After the worldwide cholera epidemic in 1993, permanent environmental monitoring of hydrographic basins was established in Pernambuco, Brazil, where cholera is endemic. After a quiescent period, 4 rfbN (serogroup O1 positive water samples that were culture negative were detected by multiplex single-tube nested PCR (MSTNPCR; 2 of these were also ctxA (cholera toxin positive. From May to June 2012, 30 V. cholerae O1 isolates were obtained by culturing samples. These isolates were analyzed for the presence of virulence genes by PCR, intergenic spacer region 16S-23S PCR (ISR-PCR, and pulsed field gel electrophoresis (PFGE. The isolates were positive for the rfbN gene and negative for the assessed pathogenic genes and were classified into 2 groups by ISR and the same profile by PFGE. Close genetic similarity was observed between them (2012 and environmental strains from 2004 to 2005, indicating the permanence of endemic V. cholerae O1 in the region.

  5. Production of Antibody Raised Against Lipopolysaccharide (LPS of Vibrio Cholerae Non-O1

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    H Shirzad

    2008-07-01

    Full Text Available Background: Cholera, an infectious disease caused by Vibrio cholerae, is primarily transmitted by ingestion of contaminated food or water. In severe cases, cholera may lead to severe dehydration, metabolic acidosis, and ultimately, hypovolemic shock and death. Methods: In this study V.cholerae non-O1 was cultured in suitable media. LPS was extracted from the surface of  bacteria by hot phenol-water method and then purified by high-speed centrifugation. For production of specific antibody against LPS, white newzeland rabbits were first immunized by whole cell bacteria and then immunized with highly purified LPS. The titre of the antiserum was determined by ELISA for each serogroup. Results: Results presented in this study indicate that serum anti-LPS antibodies raised against purified LPS of V.cholerae non-O1 can detect V.cholerae non-O1 .Conclusion: This antibody had low cross reactivity with V.cholerae O1, serotype Inaba or Ogawa. So, this antibody can be used for for detection of V. cholerae non-O1.

  6. 产色素非O1群霍乱弧菌的发现与微生物学研究%Discovery of Ten Strains of Non-O1 V. Cholerae Pigmentogens and Microbiological study

    Institute of Scientific and Technical Information of China (English)

    丁业荣; 贾利君; 时全; 解少煜; 赵本海; 常宏伟; 马振华; 汪军; 杨杰

    2000-01-01

    [目的]研究产色素非O1群霍乱弧菌在水源水中的分布、生物学性状及其致病性.[方法]在本地主要饮用水源设点、采样、培养致病性弧菌.分离、鉴定该菌过程中用营养琼脂斜面代替双糖铁琼脂取得良好效果.[结果]1994~1998年在水源水中相继检出17株产色素的革兰氏阴性弧状菌,经形态学、培养特性、生化学、血清学的系统鉴定,均符合非O1群霍乱弧菌的定义,动物实验等显示其有效强毒力.[结论]因该菌在硷性胆盐琼脂、营养琼脂斜面和血琼脂上培育后能产生水溶性褐色色素,故命名为产色素非O1群霍乱弧菌.

  7. Presence of CTX gene cluster in environmental non-O1/O139 Vibrio cholerae and its potential clinical significance

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    B Bakhshi

    2012-01-01

    Full Text Available Purpose: The aim of this study was to understand the epidemiological linkage of clinical and environmental isolates of Vibrio cholerae and to determine their genotypes and virulence genes content. Materials and Methods: A total of 60 V. cholerae strains obtained from clinical specimens (n = 40 and surface waters (n = 20 were subjected to genotyping using PFGE and determination of their virulence-associated gene clusters. Result: PCR analysis showed the presence of chromosomally located hly and RTX genetic elements in 100% and 90% of the environmental isolates, respectively. The phage-mediated genetic elements such as CTX, TLC and VPI were detected in 5% of the environmental isolates suggesting that the environmental isolates cannot acquire certain mobile gene clusters. A total of 4 and 18 pulsotypes were obtained among the clinical and environmental V. cholerae isolates, respectively. Non-pathogenic environmentally isolated V. cholerae constituted a distinct cluster with one single non-O1, non-O139 strain (EP6 carrying the virulence genes similar to the epidemic strains. This may suggest the possible potential of conversion of non-pathogenic to a pathogenic environmental strain. Conclusions: The emergence of a single environmental isolate in our study containing the pathogenicity genes amongst the diverse non-pathogenic environmental isolates needs to be further studied in the context of V. cholerae pathogenicity sero-coversion.

  8. DETECTION OF VIRULENCE GENES IN ENVIRONMENTAL STRAINS OF Vibrio cholerae FROM ESTUARIES IN NORTHEASTERN BRAZIL

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    Francisca Gleire Rodrigues de Menezes

    2014-09-01

    Full Text Available The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS, and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

  9. Detection of virulence genes in environmental strains of Vibrio cholerae from estuaries in northeastern Brazil.

    Science.gov (United States)

    Menezes, Francisca Gleire Rodrigues de; Neves, Soraya da Silva; Sousa, Oscarina Viana de; Vila-Nova, Candida Machado Vieira Maia; Maggioni, Rodrigo; Theophilo, Grace Nazareth Diogo; Hofer, Ernesto; Vieira, Regine Helena Silva dos Fernandes

    2014-01-01

    The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

  10. Non-O1/non-O139 Vibrio cholerae carrying multiple virulence factors and V. cholerae O1 in the Chesapeake Bay, Maryland.

    Science.gov (United States)

    Ceccarelli, Daniela; Chen, Arlene; Hasan, Nur A; Rashed, Shah M; Huq, Anwar; Colwell, Rita R

    2015-03-01

    Non-O1/non-O139 Vibrio cholerae inhabits estuarine and coastal waters globally, but its clinical significance has not been sufficiently investigated, despite the fact that it has been associated with septicemia and gastroenteritis. The emergence of virulent non-O1/non-O139 V. cholerae is consistent with the recognition of new pathogenic variants worldwide. Oyster, sediment, and water samples were collected during a vibrio surveillance program carried out from 2009 to 2012 in the Chesapeake Bay, Maryland. V. cholerae O1 was detected by a direct fluorescent-antibody (DFA) assay but was not successfully cultured, whereas 395 isolates of non-O1/non-O139 V. cholerae were confirmed by multiplex PCR and serology. Only a few of the non-O1/non-O139 V. cholerae isolates were resistant to ampicillin and/or penicillin. Most of the isolates were sensitive to all antibiotics tested, and 77 to 90% carried the El Tor variant hemolysin gene hlyAET, the actin cross-linking repeats in toxin gene rtxA, the hemagglutinin protease gene hap, and the type 6 secretion system. About 19 to 21% of the isolates carried the neuraminidase-encoding gene nanH and/or the heat-stable toxin (NAG-ST), and only 5% contained a type 3 secretion system. None of the non-O1/non-O139 V. cholerae isolates contained Vibrio pathogenicity island-associated genes. However, ctxA, ace, or zot was present in nine isolates. Fifty-five different genotypes showed up to 12 virulence factors, independent of the source of isolation, and represent the first report of both antibiotic susceptibility and virulence associated with non-O1/non-O139 V. cholerae from the Chesapeake Bay. Since these results confirm the presence of potentially pathogenic non-O1/non-O139 V. cholerae, monitoring for total V. cholerae, regardless of serotype, should be done within the context of public health.

  11. [Antibiotic Susceptibility of Vibrio cholerae non O1/non O139 Serogroups Isolated from Environment in the Rostov Region].

    Science.gov (United States)

    Selyanskaya, N A; Trishina, A V; Verkina, L M; Arkhangelskaya, I V; Kruglikov, V D; Zlenko, Yu M

    2014-01-01

    Analysis of the antibioticograms of 22 strains of Vibrio cholerae non O1/non O139 serogroups (ctxA- tepA-) isolated from the environment in the Rostov Region in 2011 showed that all the cultures were susceptible to ciprofloxacin, aminoglycosides, ceftriaxone, trimetoprime/sulfamethoxazole and resistant to levomycetin and furazolidone. 32%, 18% and 9% of the isolates were resistant to tetracycline, rifampicin and nalidixic acid respectively. No strains of V. cholerae susceptible to all the tested antimicrobials were detected. 37% of the V. cholerae isolates was resistant to two antibacterials and the others showed multiple resistance and contained 3-6 r-determinants of antibiotic resistance. Since the antibiotic resistance genes in Vibrio cholerae non O1/non O139 serogroups are often located on mobile genetic elements (plasmids, interferons, SXT elements), many strains of such organisms, the same as the natural environment, could serve as reservoirs of antibiotic resistance. The presence of antibiotic resistance r-determinants in the investigated strains in various combinations, the antibiotic resistance variability in the isolates collected on the same territory within a relatively short period of time require monitoring of antibiotic susceptibility in them and the use of the antibiotic for the etiotropic therapy only in strict accordance with the antibioticogram of the culture isolated from the concrete patient.

  12. Vibriosis, not cholera: toxigenic Vibrio cholerae non-O1, non-O139 infections in the United States, 1984-2014.

    Science.gov (United States)

    Crowe, S J; Newton, A E; Gould, L H; Parsons, M B; Stroika, S; Bopp, C A; Freeman, M; Greene, K; Mahon, B E

    2016-11-01

    Toxigenic strains of Vibrio cholerae serogroups O1 and O139 have caused cholera epidemics, but other serogroups - such as O75 or O141 - can also produce cholera toxin and cause severe watery diarrhoea similar to cholera. We describe 31 years of surveillance for toxigenic non-O1, non-O139 infections in the United States and map these infections to the state where the exposure probably originated. While serogroups O75 and O141 are closely related pathogens, they differ in how and where they infect people. Oysters were the main vehicle for O75 infection. The vehicles for O141 infection include oysters, clams, and freshwater in lakes and rivers. The patients infected with serogroup O75 who had food traceback information available ate raw oysters from Florida. Patients infected with O141 ate oysters from Florida and clams from New Jersey, and those who only reported being exposed to freshwater were exposed in Arizona, Michigan, Missouri, and Texas. Improving the safety of oysters, specifically, should help prevent future illnesses from these toxigenic strains and similar pathogenic Vibrio species. Post-harvest processing of raw oysters, such as individual quick freezing, heat-cool pasteurization, and high hydrostatic pressurization, should be considered.

  13. High-frequency rugose exopolysaccharide production by Vibrio cholerae strains isolated in Haiti.

    Science.gov (United States)

    Rahman, Mustafizur; Jubair, Mohammad; Alam, Meer T; Weppelmann, Thomas A; Azarian, Taj; Salemi, Marco; Sakharuk, Ilya A; Rashid, Mohammed H; Johnson, Judith A; Yasmin, Mahmuda; Morris, J Glenn; Ali, Afsar

    2014-01-01

    In October, 2010, epidemic cholera was reported for the first time in Haiti in over 100 years. Establishment of cholera endemicity in Haiti will be dependent in large part on the continued presence of toxigenic V. cholerae O1 in aquatic reservoirs. The rugose phenotype of V. cholerae, characterized by exopolysaccharide production that confers resistance to environmental stress, is a potential contributor to environmental persistence. Using a microbiologic medium promoting high-frequency conversion of smooth to rugose (S-R) phenotype, 80 (46.5%) of 172 V. cholerae strains isolated from clinical and environmental sources in Haiti were able to convert to a rugose phenotype. Toxigenic V. cholerae O1 strains isolated at the beginning of the epidemic (2010) were significantly less likely to shift to a rugose phenotype than clinical strains isolated in 2012/2013, or environmental strains. Frequency of rugose conversion was influenced by incubation temperature and time. Appearance of the biofilm produced by a Haitian clinical rugose strain (altered biotype El Tor HC16R) differed from that of a typical El Tor rugose strain (N16961R) by confocal microscopy. On whole-genome SNP analysis, there was no phylogenetic clustering of strains showing an ability to shift to a rugose phenotype. Our data confirm the ability of Haitian clinical (and environmental) strains to shift to a protective rugose phenotype, and suggest that factors such as temperature influence the frequency of transition to this phenotype.

  14. High-frequency rugose exopolysaccharide production by Vibrio cholerae strains isolated in Haiti.

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    Mustafizur Rahman

    Full Text Available In October, 2010, epidemic cholera was reported for the first time in Haiti in over 100 years. Establishment of cholera endemicity in Haiti will be dependent in large part on the continued presence of toxigenic V. cholerae O1 in aquatic reservoirs. The rugose phenotype of V. cholerae, characterized by exopolysaccharide production that confers resistance to environmental stress, is a potential contributor to environmental persistence. Using a microbiologic medium promoting high-frequency conversion of smooth to rugose (S-R phenotype, 80 (46.5% of 172 V. cholerae strains isolated from clinical and environmental sources in Haiti were able to convert to a rugose phenotype. Toxigenic V. cholerae O1 strains isolated at the beginning of the epidemic (2010 were significantly less likely to shift to a rugose phenotype than clinical strains isolated in 2012/2013, or environmental strains. Frequency of rugose conversion was influenced by incubation temperature and time. Appearance of the biofilm produced by a Haitian clinical rugose strain (altered biotype El Tor HC16R differed from that of a typical El Tor rugose strain (N16961R by confocal microscopy. On whole-genome SNP analysis, there was no phylogenetic clustering of strains showing an ability to shift to a rugose phenotype. Our data confirm the ability of Haitian clinical (and environmental strains to shift to a protective rugose phenotype, and suggest that factors such as temperature influence the frequency of transition to this phenotype.

  15. Comparative genomic analysis of two isolates of Vibrio cholerae O1 Ogawa El Tor isolated during outbreak in Mariupol in 2011.

    Science.gov (United States)

    Kuleshov, Konstantin V; Kostikova, Anna; Pisarenko, Sergey V; Kovalev, Dmitry A; Tikhonov, Sergey N; Savelievа, Irina V; Saveliev, Vilory N; Vasilieva, Oksana V; Zinich, Liliia S; Pidchenko, Nadiia N; Kulichenko, Alexander N; Shipulin, German A

    2016-10-01

    Cholera is a water-borne, severe enteric infection essentially caused by toxigenic strains of Vibrio cholera O1 and O139 serogroups. An outbreak of cholera was registered during May-July 2011 in Mariupol, Ukraine, with 33 cholera cases and 25 carriers of cholera. Following this outbreak, the toxigenic strain of V. cholerae 2011EL-301 was isolated from seawater in the recreation area of Taganrog city on the territory of Russia. The aim of our study was to understand genomic features of Mariupol isolates as well as to evaluate hypothesis about possible interconnection between the outbreak of cholera in Mariupol and the single case of isolation of V. cholerae from the Sea of Azov in Russia. Mariupol isolates were phenotypically characterized and subsequently subjected to whole genome sequencing procedure. Phylogenetic analysis based on high-quality SNPs of V. cholera O1 El Tor isolates of the 7th pandemic clade from different regions showed that clinical and environmental isolates from Mariupol outbreak were attributable to a unique phylogenetic clade within wave 3 of V. cholera O1 El Tor isolates and characterized by six clade-specific SNPs. Whereas Taganrog isolate belonged to distantly related clade which allows us to reject the hypothesis of transmission the outbreak strain of V. cholerae O1 from Ukraine to Russia in 2011. Mariupol isolates shared a common ancestor with Haiti\\Nepal-4\\India clade indicating that outbreak progenitor strain most likely originated in the South Asia region and later was introduced to Ukraine. Moreover, genomic data both based on hqSNPs and similarity of virulence-associated mobile genomic elements of Mariupol isolates suggests that environmental and clinical isolates are a part of joint outbreak which confirms the role of contaminated domestic sewage, as an element of the complex chain of infection spread during cholera outbreak. In general, the genome-wide comparative analysis of both genes and genomic regions of epidemiological

  16. Obtaining of a rapid diagnostic test for Cholera, based on latex particles coupled with a monoclonal antibody against Vibrio cholera O1 lipopolysaccharide

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    Fátima Reyes-López

    2015-08-01

    Full Text Available Cholera is an acute contagious intestinal disease caused by ingestion of food or water contaminated with O1 and O139 serotypes of the bacterium Vibrio cholerae. Cholera is characterized by abundant secretory diarrhea leading to dehydration. Death occurs within hours without treatment, so early diagnosis is very important, especially at the beginning of the disease, because it is difficult to differentiate from other acute diarrheal diseases. The diagnostic golden test is the stool culture; however, it does not guarantee a rapid detection of the disease. Rapid tests have been recently developed; they are based on test strips and agglutination with latex particles, which are very effective, but difficult to acquire for their high prices. The objective of this research was to obtain a quick assay based on latex particles coupled with a monoclonal antibody (mAb against V. cholerae O1 lipopolysaccharide obtained in Finlay Institute. Latex particles of 0.8 µm were used in a 10% suspension, and they were coupled to the mAb (0.25 mg/ml for 2 hours at 37°C. The sensitivity, specificity and performance were evaluated in 84 stool samples from patients with presumptive diagnosis of cholera. The diagnostic test obtained showed no cross-reactivity against no-O1 strains and other enteropathogens. Latex diagnostic test showed values of sensitivity, specificity and efficacy of 97.87; 97.29 and 97.6% respectively, very similar to the commercial diagnostic test CTK- Biotech. The latex reagent obtained can be used in the rapid diagnosis of the disease.

  17. Plasmid mediated antibiotic resistance ofVibrio cholerae O1 biotype El Tor serotype Ogawa associated with an outbreak in Kolkata, India

    Institute of Scientific and Technical Information of China (English)

    Shyamapada Mandal; Manisha DebMandal; Nishith Kumar Pal

    2010-01-01

    Objective:To determine the antibiotic resistance ofVibrio cholerae (V. cholerae)O1 biotype El Tor serotype Ogawa isolates involved in an outbreak of watery diarrhea in Kolkata, and to explore the role of plasmid in mediating antibiotic resistance.Methods: Antibiotic susceptibility and minimum inhibitory concentration(MIC) values of antibiotics for the isolated V. choleraeO1 Ogawa (n=12) were determined by disk diffusion and agar dilution methods, respectively, using ampicillin (Am), chloramphenicol (C), trimethoprim (Tm), tetracycline (T), erythromycine (Er), nalidixic acid (Nx), ciprofloxacin (Cp), amikacin (Ak) and cefotaxime (Cf). Plasmid curing of multidrug resistant(MDR)V. choleraeO1 Ogawa strains was done following ethidium bromide treatment. Following electrophoresis, the plasmidDNAs, extracted from the isolatedMDRV. choleraeO1 Ogawa strains and their cured derivatives, were visualized and documented in‘gel doc’ system.Results: The outbreak causingV. choleraeO1 Ogawa isolates wereMDR as determined by disk diffusion susceptibility test, andMIC determination. The isolates showed three different drug resistance patterns: AmTmTErNx (for6 isolates), TmTErCp (for 5 isolates), and AmTmNx (for one isolate), and showed uniform sensitivity to C, Ak and Cf. The loss of plasmids with the concomitant loss of resistance to Am, Tm, T and Er of the isolates occurred following ethidium bromide treatment.Conclusions: The current findings suggest that theV. choleraeO1Ogawa associated with the cholera outbreak wereMDR, and resistance to Am, Tm, T and Er among the isolates were plasmid mediated.

  18. A recent outbreak of cholera due to Vibrio cholerae O1 Ogawa in & around Chandigarh, North India.

    Science.gov (United States)

    Taneja, Neelam; Kaur, Jasjit; Sharma, Kusum; Singh, Malkit; Kalra, J K; Sharma, N M; Sharma, Meera

    2003-06-01

    An outbreak of cholera caused by Vibrio cholerae O1 Ogawa occurred in and around Chandigarh during July 22-31, 2002. Of the 303 patients admitted to two hospitals, 82 were confirmed by culture. Two rehabilitation colonies located at the periphery of Chandigarh were mainly affected. The isolates were biotyped as Eltor and were susceptible to many antibiotics. Thirty one (35.2%) of 88 water samples showed evidence of faecal contamination. The survey of the area revealed sewage contamination of the drinking water supply. The outbreak was controlled by providing safe drinking water to the people and correcting the defects in the sewage and water pipelines.

  19. Isolation frequency and susceptibility pattern of non-O1 and non-O139 Vibrio cholerae in a tertiary health care laboratory, 1999-2012.

    Science.gov (United States)

    Irfan, S; Fasih, N; Ghanchi, N K; Khan, E

    2016-04-28

    In the past decade the importance of non-O1 and non-O139 strains of Vibrio cholerae has been highlighted globally. This study aimed to evaluate the frequency and antimicrobial susceptibility profile of non-O1 and non-O139 V. cholerae in Pakistan. Data of stool specimens yielding growth of non-O1 and non-O139 V. cholerae isolated at a national referral laboratory from 1999 to 2012 were retrospectively analysed and evaluated for resistance to ampicillin, tetracycline, chloramphenicol, co-trimoxazole and ofloxacin. A total of 95 800 stool samples submitted over 1999-2012 yielded 3668 strains of V. cholerae, of which 6% were non-O1 and non-O139 V. cholerae. A high isolation rate was found in the summer season, with a peak in the year 2003. Antimicrobial susceptibility data revealed increasing resistance to co-trimoxazole and ampicillin, but strains remained highly susceptible to ofloxacin. Active surveillance of serotypes and antimicrobial susceptibility is essential to predict future epidemics and define measures to curtail the disease.

  20. Increased isolation frequency of toxigenic Vibrio cholerae O1 from environmental monitoring sites in Haiti.

    Science.gov (United States)

    Alam, Meer T; Weppelmann, Thomas A; Longini, Ira; De Rochars, Valery Madsen Beau; Morris, John Glenn; Ali, Afsar

    2015-01-01

    Since the identification of the first cholera case in 2010, the disease has spread in epidemic form throughout the island nation of Haiti; as of 2014, about 700,000 cholera cases have been reported, with over 8,000 deaths. While case numbers have declined, the more fundamental question of whether the causative bacterium, Vibrio cholerae has established an environmental reservoir in the surface waters of Haiti remains to be elucidated. In a previous study conducted between April 2012 and March 2013, we reported the isolation of toxigenic V. cholerae O1 from surface waters in the Ouest Department. After a second year of surveillance (April 2013 to March 2014) using identical methodology, we observed a more than five-fold increase in the number of water samples containing culturable V. cholerae O1 compared to the previous year (1.7% vs 8.6%), with double the number of sites having at least one positive sample (58% vs 20%). Both seasonal water temperatures and precipitation were significantly related to the frequency of isolation. Our data suggest that toxigenic V. cholerae O1 are becoming more common in surface waters in Haiti; while the basis for this increase is uncertain, our findings raise concerns that environmental reservoirs are being established.

  1. Distribution and content of class 1 integrons in different Vibrio cholerae O-serotype strains isolated in Thailand

    DEFF Research Database (Denmark)

    Dalsgaard, Anders; Forslund, Anita; Serichantalergs, Oralak

    2000-01-01

    In this study, 176 clinical and environmental Vibrio cholerae strains of different O serotypes isolated in Thailand from 1982 to 1995 were selected and studied for the presence of class 1 integrons, a new group of genetic elements which carry antibiotic resistance genes. Using PCR and DNA...... by the strains. Serotype O139 strains did not contain class 1 integrons. However, the appearance and disappearance of the O139 serotype in the coastal city Samutsakorn in 1992 and 1993 were associated with the emergence of a distinct V. cholerae O1 strain which contained the aad-V resistance gene cassette. A 150....... cholerae O serotypes of mainly clinical origin in Thailand....

  2. Detection of ctx gene positive non-O1/non-O139 V. cholerae in shrimp aquaculture environments.

    Science.gov (United States)

    Madhusudana, Rao B; Surendran, P K

    2013-06-01

    Water and post-larvae samples from black tiger (Penaeus monodon) shrimp hatcheries; pond water, pond sediment and shrimp from aquaculture farms were screened for the presence of V. cholerae. A V. cholerae-duplex PCR method was developed by utilizing V. cholerae species specific sodB primers and ctxAB genes specific primers. Incidence of V. cholerae was not observed in shrimp hatchery samples but was noticed in aquaculture samples. The incidence of V. cholerae was higher in pond water (7.6%) than in pond sediment (5.2%). Shrimp head (3.6%) portion had relatively higher incidence than shrimp muscle (1.6%). All the V. cholerae isolates (n = 42) belonged to non-O1/non-O139 serogroup, of which 7% of the V. cholerae isolates were potentially cholera-toxigenic (ctx positive). All the ctx positive V. cholerae (n = 3) were isolated from the pond water. Since, cholera toxin (CT) is the major contributing factor for cholera gravis, it is proposed that the mere presence of non-O1/non-O139 V. cholerae need not be the biohazard criterion in cultured black tiger shrimp but only the presence of ctx carrying non-O1/non-O139 V. cholerae may be considered as potential public health risk.

  3. Sustained Local Diversity of Vibrio cholerae O1 Biotypes in a Previously Cholera-Free Country

    Directory of Open Access Journals (Sweden)

    Yan Boucher

    2016-07-01

    Full Text Available Although the current cholera pandemic can trace its origin to a specific time and place, many variants of Vibrio cholerae have caused this disease over the last 50 years. The relative clinical importance and geographical distribution of these variants have changed with time, but most remain in circulation. Some countries, such as Mexico and Haiti, had escaped the current pandemic, until large epidemics struck them in 1991 and 2010, respectively. Cholera has been endemic in these countries ever since. A recent retrospective study in mBio presents the results of more than 3 decades of V. cholerae monitoring from environmental and clinical sources in Mexico (S. Y. Choi et al., mBio 7:e02160-15, 2016, http://dx.doi.org/10.1128/mBio.02160-15. It reveals that multiple V. cholerae variants, including classical strains from the previous pandemic, as well as completely novel biotypes, have been circulating in Mexico. This discovery has important implications for the epidemiology and evolution of V. cholerae.

  4. Biochemical and full genome sequence analyses of clinical Vibrio cholerae isolates in Mexico reveals the presence of novel V. cholerae strains.

    Science.gov (United States)

    Díaz-Quiñonez, José Alberto; Hernández-Monroy, Irma; Montes-Colima, Norma Angélica; Moreno-Pérez, María Asunción; Galicia-Nicolás, Adriana Guadalupe; López-Martínez, Irma; Ruiz-Matus, Cuitláhuac; Kuri-Morales, Pablo; Ortíz-Alcántara, Joanna María; Garcés-Ayala, Fabiola; Ramírez-González, José Ernesto

    2016-05-01

    The first week of September 2013, the National Epidemiological Surveillance System identified two cases of cholera in Mexico City. The cultures of both samples were confirmed as Vibrio cholerae serogroup O1, serotype Ogawa, biotype El Tor. Initial analyses by PFGE and by PCR-amplification of the virulence genes, suggested that both strains were similar, but different from those previously reported in Mexico. The following week, four more cases were identified in a community in the state of Hidalgo, located 121 km northeast of Mexico City. Thereafter a cholera outbreak started in the region of La Huasteca. Genomic analyses of the four strains obtained in this study confirmed the presence of Pathogenicity Islands VPI-1 and -2, VSP-1 and -2, and of the integrative element SXT. The genomic structure of the 4 isolates was similar to that of V. cholerae strain 2010 EL-1786, identified during the epidemic in Haiti in 2010.

  5. Major Shift of Toxigenic V. cholerae O1 from Ogawa to Inaba Serotype Isolated from Clinical and Environmental Samples in Haiti

    Science.gov (United States)

    Alam, Meer T.; Ray, Shrestha S.; Chun, Camille N.; Chowdhury, Zahara G.; Rashid, Mohammed H.; Madsen Beau De Rochars, Valery E.; Ali, Afsar

    2016-01-01

    In October of 2010, an outbreak of cholera was confirmed in Haiti for the first time in more than a century. A single clone of toxigenic Vibrio cholerae O1 biotype El Tor serotype Ogawa strain was implicated as the cause. Five years after the onset of cholera, in October, 2015, we have discovered a major switch (ranging from 7 to 100%) from Ogawa serotype to Inaba serotype. Furthermore, using wbeT gene sequencing and comparative sequence analysis, we now demonstrate that, among 2013 and 2015 Inaba isolates, the wbeT gene, responsible for switching Ogawa to Inaba serotype, sustained a unique nucleotide mutation not found in isolates obtained from Haiti in 2012. Moreover, we show that, environmental Inaba isolates collected in 2015 have the identical mutations found in the 2015 clinical isolates. Our data indicate that toxigenic V. cholerae O1 serotype Ogawa can rapidly change its serotype to Inaba, and has the potential to cause disease in individuals who have acquired immunity against Ogawa serotype. Our findings highlight the importance of monitoring of toxigenic V. cholerae O1 and cholera in countries with established endemic disease. PMID:27716803

  6. Vibrio cholerae strains possess multiple strategies for abiotic and biotic surface colonization.

    Science.gov (United States)

    Mueller, Ryan S; McDougald, Diane; Cusumano, Danielle; Sodhi, Nidhi; Kjelleberg, Staffan; Azam, Farooq; Bartlett, Douglas H

    2007-07-01

    Despite its notoriety as a human pathogen, Vibrio cholerae is an aquatic microbe suited to live in freshwater, estuarine, and marine environments where biofilm formation may provide a selective advantage. Here we report characterization of biofilms formed on abiotic and biotic surfaces by two non-O1/O139 V. cholerae strains, TP and SIO, and by the O1 V. cholerae strain N16961 in addition to the isolation of 44 transposon mutants of SIO and TP impaired in biofilm formation. During the course of characterizing the mutants, 30 loci which have not previously been associated with V. cholerae biofilms were identified. These loci code for proteins which perform a wide variety of functions, including amino acid metabolism, ion transport, and gene regulation. Also, when the plankton colonization abilities of strains N16961, SIO, and TP were examined, each strain showed increased colonization of dead plankton compared with colonization of live plankton (the dinoflagellate Lingulodinium polyedrum and the copepod Tigriopus californicus). Surprisingly, most of the biofilm mutants were not impaired in plankton colonization. Only mutants impaired in motility or chemotaxis showed reduced colonization. These results indicate the presence of both conserved and variable genes which influence the surface colonization properties of different V. cholerae subspecies.

  7. Genomic profile of antibiotic resistant, classical ctxB positive Vibrio cholerae O1 biotype El Tor isolated in 2003 and 2005 from Puri, India: A retrospective study

    Directory of Open Access Journals (Sweden)

    T Bhotra

    2016-01-01

    Full Text Available Objectives: To examine eight strains of Vibrio cholerae O1 isolated in 2003 and 2005 from Puri, India, for antibiotic susceptibility, presence of virulence and regulatory genes, cholera toxin (CT production, CTX arrangement and genomic profiles. Materials and Methods: Bacterial strains were tested for antibiotic susceptibility using disc diffusion assay. Polymerase chain reaction determined the presence of antibiotic resistance, virulence and regulatory genes. To determine the type of cholera toxin subunit B (ctxB, nucleotide sequencing was performed. Southern hybridisation determined the number and arrangement of CTXΦ. Ribotyping and pulsed-field gel electrophoresis (PFGE were used to determine the genomic profile of isolates. Results: All the eight strains, except one strain, showed resistant to nalidixic acid, sulphamethoxazole, streptomycin and trimethoprim and possessed the sullI, strB, dfrA1 and int SXT genes. All the strains carried the toxin-co-regulated pilus pathogenicity island, the CTX genetic element, the repeat in toxin and produced CT. Restriction fragment length polymorphism (RFLP analysis showed that V. cholerae O1 possess a single copy of the CTX element flanked by tandemly arranged RS element. Nucleotide sequencing of the ctxB gene showed the presence of classical ctxB. RFLP analysis of conserved rRNA gene showed two ribotype patterns. PFGE analysis also showed at least three PFGE patterns, irrespective of year of isolations, indicating the genomic relatedness among them. Conclusion: Overall, these data suggest that classical ctxB-positive V. cholerae O1 El Tor strains that appeared in 2003 continue to cause infection in 2005 in Puri, India, and belong to identical ribotype(s and/or pulsotype(s. There is need to continuous monitor the emergence of variant of El Tor because it will improve our understanding of the evolution of new clones of variant of V. cholerae.

  8. O Serogroup-Specific Touchdown-Multiplex Polymerase Chain Reaction for Detection and Identification of Vibrio cholerae O1, O139, and Non-O1/Non-O139

    Directory of Open Access Journals (Sweden)

    Adisak Bhumiratana

    2014-01-01

    Full Text Available A novel, sensitive locus-specific touchdown-multiplex polymerase chain reaction (TMPCR, which is based on two-stage amplification pertaining to multiplex PCR and conditional touchdown strategy, was used in detecting and differentiating Vibrio cholerae serogroups. A panel of molecular marker-based TMPCR method generates reproducible profiles of V. cholerae-specific (588 bp amplicons derived from ompW gene encoding the outer membrane protein and serogroup-specific amplicons, 364 bp for the O1 and 256 bp for the O139, authentically copied from rfb genes responsible for the lipopolysaccharide biosynthesis. The TMPCR amplification efficiency yields either equally or unequally detectable duplex DNA bands of the O1 (588 and 364 bp and O139 (588 and 256 bp or a DNA fragment of non-O1/non-O139 (588 bp while providing no false positive identifications using the genomic DNA templates of the other vibrios and Enterobacteriaceae. The reciprocal analysis of two-template combinations demonstrated that, using V. cholerae O1, O139, or equally mixed O1 and O139, the TMPCR had a detection limit of as low as 100 pg of the O1, O139, or non-O1/non-O139 in reactions containing unequally or equally mixed gDNAs. In addition, the O serogroup-specific TMPCR method had 100% agreement with the serotyping method when examined for the serotyped V. cholerae reference strains and those recovered from clinical samples. The potential benefit of using this TMPCR tool would augment the serotyping method used in epidemiological surveillance and monitoring of V. cholerae serogroups, O1, O139, and non-O1/non-O139 present in clinical and environmental samples.

  9. Phase variable O antigen biosynthetic genes control expression of the major protective antigen and bacteriophage receptor in Vibrio cholerae O1.

    Directory of Open Access Journals (Sweden)

    Kimberley D Seed

    2012-09-01

    Full Text Available The Vibrio cholerae lipopolysaccharide O1 antigen is a major target of bacteriophages and the human immune system and is of critical importance for vaccine design. We used an O1-specific lytic bacteriophage as a tool to probe the capacity of V. cholerae to alter its O1 antigen and identified a novel mechanism by which this organism can modulate O antigen expression and exhibit intra-strain heterogeneity. We identified two phase variable genes required for O1 antigen biosynthesis, manA and wbeL. manA resides outside of the previously recognized O1 antigen biosynthetic locus, and encodes for a phosphomannose isomerase critical for the initial step in O1 antigen biosynthesis. We determined that manA and wbeL phase variants are attenuated for virulence, providing functional evidence to further support the critical role of the O1 antigen for infectivity. We provide the first report of phase variation modulating O1 antigen expression in V. cholerae, and show that the maintenance of these phase variable loci is an important means by which this facultative pathogen can generate the diverse subpopulations of cells needed for infecting the host intestinal tract and for escaping predation by an O1-specific phage.

  10. Molecular characterization of high-level-cholera-toxin-producing El Tor variant Vibrio cholerae strains in the Zanzibar Archipelago of Tanzania.

    Science.gov (United States)

    Naha, A; Chowdhury, G; Ghosh-Banerjee, J; Senoh, M; Takahashi, T; Ley, B; Thriemer, K; Deen, J; Seidlein, L V; Ali, S M; Khatib, A; Ramamurthy, T; Nandy, R K; Nair, G B; Takeda, Y; Mukhopadhyay, A K

    2013-03-01

    Analysis of 1,180 diarrheal stool samples in Zanzibar detected 247 Vibrio cholerae O1, Ogawa strains in 2009. Phenotypic traits and PCR-based detection of rstR, rtxC, and tcpA alleles showed that they belonged to the El Tor biotype. Genetic analysis of ctxB of these strains revealed that they were classical type, and production of classical cholera toxin B (CTB) was confirmed by Western blotting. These strains produced more CT than the prototype El Tor and formed a separate cluster by pulsed-field gel electrophoresis (PFGE) analysis.

  11. Survivability of Vibrio cholerae O1 in Cooked Rice, Coffee, and Tea

    Directory of Open Access Journals (Sweden)

    John Yew Huat Tang

    2013-01-01

    Full Text Available This study aimed to investigate the survival of Vibrio cholerae O1 in 3 types of preparation for cooked rice, Oryza sativa L., (plain rice, rice with coconut milk, and rice with ginger; coffee, Coffea canephora, (plain coffee, coffee with sugar, and coffee with sweetened condensed milk; and tea, Camellia sinensis, (plain tea, tea with sugar, and tea with sweetened condensed milk held at room temperature (27°C. The survival of V. cholerae O1 was determined by spread plate method on TCBS agar. Initial cultures of 8.00 log CFU/mL were inoculated into each food sample. After 6 h incubation, significant growth was only detected in rice with coconut milk (9.67 log CFU/mL; P<0.05. However, all 3 types of rice preparation showed significant growth of V. cholerae after 24 h (P<0.05. For coffee and tea preparations, V. cholerae survived up to 6 h in tea with condensed milk (4.72 log CFU/mL but not in similar preparation of coffee. This study showed evidence for the survivability of V. cholerae in rice, coffee, and tea. Thus, holding these food and beverages for an extended period of time at room temperature should be avoided.

  12. Label-free electrochemical immunosensor based on cerium oxide nanowires for Vibrio cholerae O1 detection

    Energy Technology Data Exchange (ETDEWEB)

    Tam, Phuong Dinh, E-mail: phuongdinhtam@gmail.com; Thang, Cao Xuan, E-mail: thang.caoxuan@hust.edu.vn

    2016-01-01

    This paper developed a label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application. The CeO{sub 2} nanowires were synthesized by hydrothermal reaction. The immobilization of Anti-V. cholerae O1 onto CeO{sub 2} nanowire-deposited sensor was performed via an amino ester, which was created by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and sulfo-N-hydroxysuccinimide. The electrochemical responses of the immunosensor were studied by electrochemical impedance spectroscopy with [Fe (CN) {sub 6}] {sup 3−/4−} as redox probe. A linear response in electron transfer resistance for cell of V. cholerae O1 concentration was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. The immunosensor sensitivity was 56.82 Ω/CFU·mL{sup −1}. Furthermore, the parameters affecting immunosensor response were also investigated, as follows: pH value, immunoreaction time, incubation temperature, and anti-V. cholerae O1 concentration. - Highlights: • A label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application was developed. • A linear response was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. • The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. • The immunosensor sensitivity was 56.82 Ω/CFU.mL{sup −1}.

  13. Development of lipopolysaccharide-mimicking peptides and their immunoprotectivity against Vibrio cholerae serogroup O1.

    Science.gov (United States)

    Mohammad Pour Ghazi, Fatemeh; Gargari, Seyed Latif Mousavi

    2016-11-01

    Vibrio cholerae serogroup O1 is the main causative agent of cholera diseases defined by life threatening rice watery diarrhea. Cholera routine vaccination has failed in controlling epidemics in developing countries because of their hard and expensive production. In this study, our aim was to investigate phage displayed mimotopes that could mimic V. cholerae lipopolysaccharide (LPS). Although LPS of Vibrio, as an endotoxin, can stimulate the immune system, thereby making it a suitable candidate for cholera vaccine, its toxicity remains as a main problem. Phage particles displaying 12 amino acid peptides were selected from phage library mimicking the antigenic epitopes of LPS from vibrio. The screening was carried out using single-domain antibody fragment VHH against LPS as target through three rounds of selection. Three clones with highest affinity to VHH were selected. To find out a new and efficient vaccine against cholera, these three phage particles containing high-affinity peptides were administered to mice to investigate the active and passive immunity. Out of 20 particles, three showed the highest affinity toward VHH. ELISA was carried out with immunized mice sera using LPS and three selected phages particles individually. ETEC, Shigella sonnei, and clinical isolates were used as bacterial targets. These three selected phages (individually or in combination) could stimulate mice immune system producing active and passive immunity. The mice immunized with phage particles could protect about 14 LD50 of V. cholerae. In conclusion, these peptides are mimicking LPS and can potentially act as vaccine candidates against V. cholerae. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

  14. Transcutaneous immunization with toxin-coregulated pilin A induces protective immunity against Vibrio cholerae O1 El Tor challenge in mice.

    Science.gov (United States)

    Rollenhagen, Julianne E; Kalsy, Anuj; Cerda, Francisca; John, Manohar; Harris, Jason B; Larocque, Regina C; Qadri, Firdausi; Calderwood, Stephen B; Taylor, Ronald K; Ryan, Edward T

    2006-10-01

    Toxin-coregulated pilin A (TcpA) is the main structural subunit of a type IV bundle-forming pilus of Vibrio cholerae, the cause of cholera. Toxin-coregulated pilus is involved in formation of microcolonies of V. cholerae at the intestinal surface, and strains of V. cholerae deficient in TcpA are attenuated and unable to colonize intestinal surfaces. Anti-TcpA immunity is common in humans recovering from cholera in Bangladesh, and immunization against TcpA is protective in murine V. cholerae models. To evaluate whether transcutaneously applied TcpA is immunogenic, we transcutaneously immunized mice with 100 mug of TcpA or TcpA with an immunoadjuvant (cholera toxin [CT], 50 mug) on days 0, 19, and 40. Mice immunized with TcpA alone did not develop anti-TcpA responses. Mice that received transcutaneously applied TcpA and CT developed prominent anti-TcpA immunoglobulin G (IgG) serum responses but minimal anti-TcpA IgA. Transcutaneous immunization with CT induced prominent IgG and IgA anti-CT serum responses. In an infant mouse model, offspring born to dams transcutaneously immunized either with TcpA and CT or with CT alone were challenged with 10(6) CFU (one 50% lethal dose) wild-type V. cholerae O1 El Tor strain N16961. At 48 h, mice born to females transcutaneously immunized with CT alone had 36% +/- 10% (mean +/- standard error of the mean) survival, while mice born to females transcutaneously immunized with TcpA and CT had 69% +/- 6% survival (P < 0.001). Our results suggest that transcutaneous immunization with TcpA and an immunoadjuvant induces protective anti-TcpA immune responses. Anti-TcpA responses may contribute to an optimal cholera vaccine.

  15. Pathophysiological mechanisms of diarrhea caused by the Vibrio cholerae O1 El Tor variant: an in vivo study in mice.

    Science.gov (United States)

    Satitsri, Saravut; Pongkorpsakol, Pawin; Srimanote, Potjanee; Chatsudthipong, Varanuj; Muanprasat, Chatchai

    2016-10-01

    Cholera is caused by infection with Vibrio cholerae. This study aimed to investigate the pathophysiology of diarrhea caused by the V. cholerae O1 El Tor variant (EL), a major epidemic strain causing severe diarrhea in several regions. In the ligated ileal loop model of EL-induced diarrhea in the ICR mice, a cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor and a calcium-activated chloride channel (CaCC) inhibitor similarly inhibited intestinal fluid secretion. In addition, barrier disruption and NF-κB-mediated inflammatory responses, e.g., iNOS and COX-2 expression, were observed in the infected ileal loops. Interestingly, intestinal fluid secretion and barrier disruption were suppressed by NF-κB and COX-2 inhibitors, whereas an iNOS inhibitor suppressed barrier disruption without affecting fluid secretion. Furthermore, EP2 and EP4 PGE2 receptor antagonists ameliorated the fluid secretion in the infected ileal loops. The amount of cholera toxin (CT) produced in the ileal loops by the EL was ∼2.4-fold of the classical biotype. The CT transcription inhibitor virstatin, a toll-like receptor-4 (TLR-4) antibody and a CT antibody suppressed the EL-induced intestinal fluid secretion, barrier disruption and COX-2 expression. The CT at levels detected during EL infection induced mild intestinal barrier disruption without inducing inflammatory responses in mouse intestine. Collectively, this study indicates that CT-induced intestinal barrier disruption and subsequent TLR-4-NF-κB-mediated COX-2 expression are involved in the pathogenesis of EL-induced diarrhea and represent promising novel therapeutic targets of cholera.

  16. New Vibrio cholerae O1 Biotype ElTor bacteriophages

    Directory of Open Access Journals (Sweden)

    Sen Anindito

    2005-04-01

    Full Text Available Abstract We report the presence of three new O1 ElTor vibriophages named AS1, AS2 and AS3, isolated from the sewage and pond waters of the outskirts of Kolkata. A few phages, named AS4, with hexagonal heads and abnormally long tails with typical curly projections were also found in the water samples.

  17. Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India.

    Science.gov (United States)

    Das, Moon Moon; Bhotra, Tilothama; Zala, Dolatsinh; Singh, Durg Vijai

    2016-08-01

    Vibrio cholerae O1 biotype El Tor, the causative agent of the seventh pandemic, has recently been replaced by strains carrying classical and Haitian ctxB in India, Haiti and other parts of the world. We conducted phenotypic and genetic tests to characterize V. cholerae O1 isolated between 2012 and 2014 from Silvassa, India, to examine the presence of virulence and regulatory genes, seventh pandemic marker, ctxB type and biofilm formation and to study genomic diversity. Of the 59 V. cholerae O1, eight isolates belong to El Tor prototype, one to classical prototype and the remaining isolates have attributes of both classical and El Tor biotypes. PCR and ctxB gene sequencing revealed the presence of classical ctxB in four strains and Haitian ctxB in 55 isolates; indicating that isolates were either an El Tor or hybrid variant. All isolates carried virulence, regulatory, adherence, Vibrio seventh pandemic pathogenicity island I and seventh pandemic group-specific marker VC2346, in addition to tcpAET and rstRET, the features of seventh pandemic strains, and produced cholera toxin and biofilm. PFGE analysis showed that the majority of isolates are clonal and belong to fingerprint pattern A; however, pattern B is unrelated and patterns C and D are distinct, suggesting considerable diversity in the genomic content among them. These data thus show that isolates from Silvassa are genetically diverse and that Haitian ctxB and hybrid phenotypes are undergoing global dissemination.

  18. Survey on antimicrobial resistance patterns in Vibrio vulnificus and Vibrio cholerae non-O1/non-O139 in Germany reveals carbapenemase-producing Vibrio cholerae in coastal waters.

    Science.gov (United States)

    Bier, Nadja; Schwartz, Keike; Guerra, Beatriz; Strauch, Eckhard

    2015-01-01

    An increase in the occurrence of potentially pathogenic Vibrio species is expected for waters in Northern Europe as a consequence of global warming. In this context, a higher incidence of Vibrio infections is predicted for the future and forecasts suggest that people visiting and living at the Baltic Sea are at particular risk. This study aimed to investigate antimicrobial resistance patterns among Vibrio vulnificus and Vibrio cholerae non-O1/non-O139 isolates that could pose a public health risk. Antimicrobial susceptibility of 141 V. vulnificus and 184 V. cholerae non-O1/non-O139 strains isolated from German coastal waters (Baltic Sea and North Sea) as well as from patients and retail seafood was assessed by broth microdilution and disk diffusion. Both species were susceptible to most of the agents tested (12 subclasses) and no multidrug-resistance was observed. Among V. vulnificus isolates, non-susceptibility was exclusively found toward aminoglycosides. In case of V. cholerae, a noticeable proportion of strains was non-susceptible to aminopenicillins and aminoglycosides. In addition, resistance toward carbapenems, quinolones, and folate pathway inhibitors was sporadically observed. Biochemical testing indicated the production of carbapenemases with unusual substrate specificity in four environmental V. cholerae strains. Most antimicrobial agents recommended for treatment of V. vulnificus and V. cholerae non-O1/non-O139 infections were found to be effective in vitro. However, the occurrence of putative carbapenemase producing V. cholerae in German coastal waters is of concern and highlights the need for systematic monitoring of antimicrobial susceptibility in potentially pathogenic Vibrio spp. in Europe.

  19. Genetic and phenotypic analysis of Vibrio cholerae non-O1, non-O139 isolated from German and Austrian patients.

    Science.gov (United States)

    Schirmeister, F; Dieckmann, R; Bechlars, S; Bier, N; Faruque, S M; Strauch, E

    2014-05-01

    Vibrio cholerae belonging to the non-O1, non-O139 serogroups are present in the coastal waters of Germany and in some German and Austrian lakes. These bacteria can cause gastroenteritis and extraintestinal infections, and are transmitted through contaminated food and water. However, non-O1, non-O139 V. cholerae infections are rare in Germany. We studied 18 strains from German and Austrian patients with diarrhea or local infections for their virulence-associated genotype and phenotype to assess their potential for infectivity in anticipation of possible climatic changes that could enhance the transmission of these pathogens. The strains were examined for the presence of genes encoding cholera toxin and toxin-coregulated pilus (TCP), as well as other virulence-associated factors or markers, including hemolysins, repeats-in-toxin (RTX) toxins, Vibrio seventh pandemic islands VSP-1 and VSP-2, and the type III secretion system (TTSS). Phenotypic assays for hemolysin activity, serum resistance, and biofilm formation were also performed. A dendrogram generated by incorporating the results of these analyses revealed genetic differences of the strains correlating with their clinical origin. Non-O1, non-O139 strains from diarrheal patients possessed the TTSS and/or the multifunctional autoprocessing repeats-in-toxin (MARTX) toxin, which were not found in the strains from ear or wound infections. Routine matrix-assisted laser desorption/ionization (MALDI-TOF) mass spectrometry (MS) analysis of all strains provided reliable identification of the species but failed to differentiate between strains or clusters. The results of this study indicate the need for continued surveillance of V. cholerae non-O1, non-O139 in Germany, in view of the predicted increase in the prevalence of Vibrio spp. due to the rise in surface water temperatures.

  20. Validation and characterization of a human volunteer challenge model for cholera by using frozen bacteria of the new Vibrio cholerae epidemic serotype, O139

    NARCIS (Netherlands)

    Cohen, MB; Giannella, RA; Losonsky, GA; Lang, DR; Parker, S; Hawkins, JA; Gunther, C; Schiff, GA

    1999-01-01

    Until recently, all epidemic strains of Vibrio cholerae were of the O1 serotype. Current epidemics have also been caused by a new serotype, Vibrio cholerae O139. Although the pathogenesis and clinical features of O139 cholera are similar to those of O1 cholera, immunity to serotype O1 does not confe

  1. Phenotypic Analysis Reveals that the 2010 Haiti Cholera Epidemic Is Linked to a Hypervirulent Strain.

    Science.gov (United States)

    Satchell, Karla J F; Jones, Christopher J; Wong, Jennifer; Queen, Jessica; Agarwal, Shivani; Yildiz, Fitnat H

    2016-09-01

    Vibrio cholerae O1 El Tor strains have been responsible for pandemic cholera since 1961. These strains have evolved over time, spreading globally in three separate waves. Wave 3 is caused by altered El Tor (AET) variant strains, which include the strain with the signature ctxB7 allele that was introduced in 2010 into Haiti, where it caused a devastating epidemic. In this study, we used phenotypic analysis to compare an early isolate from the Haiti epidemic to wave 1 El Tor isolates commonly used for research. It is demonstrated that the Haiti isolate has increased production of cholera toxin (CT) and hemolysin, increased motility, and a reduced ability to form biofilms. This strain also outcompetes common wave 1 El Tor isolates for colonization of infant mice, indicating that it has increased virulence. Monitoring of CT production and motility in additional wave 3 isolates revealed that this phenotypic variation likely evolved over time rather than in a single genetic event. Analysis of available whole-genome sequences and phylogenetic analyses suggested that increased virulence arose from positive selection for mutations found in known and putative regulatory genes, including hns and vieA, diguanylate cyclase genes, and genes belonging to the lysR and gntR regulatory families. Overall, the studies presented here revealed that V. cholerae virulence potential can evolve and that the currently prevalent wave 3 AET strains are both phenotypically distinct from and more virulent than many El Tor isolates.

  2. GENOTYPE AND DRUG RESISTANCE OF CLINICAL AND ENVIRONMENTAL VIBRIO CHOLERAE NON-O1/NON-O139 IN NORTHEASTERN THAILAND.

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    Chomvarin, Chariya; Jumroenjit, Warin; Tangkanakul, Waraluk; Hasan, Nur A; Chaicumpar, Kunyaluk; Faksri, Kiatichai; Huq, Anwar

    2014-11-01

    A total of 124 V cholerae non-O1/non-O139 isolates were collected in Khon Kaen, Thailand from diarrheal patients, asymptomatic carriers and environmental water. The presence of virulence-associated and regulatory genes including ctxA, tcpA, zot, ace, ompU, stn, hlyA and toxR) were examined using multiplex PCR. The genomic diversity of the various V. cholerae isolates were differentiated using the random amplified polymorphic DNA (RAPD) method. Antimicrobial susceptibility was tested using disk diffusion. All of V. cholerae non-O/non-O139 isolates carried hlyA and toxR and none carried ctxA and tcpA. The zot, ace and both genes together were found in 1.6%, 4.7% and 4.7% of 64 clinical V. cholerae non-O1 isolates, respectively, while the environmental ones did not. The stn gene was found in 3.1% (2/64) of the clinical and 3.3% (2/60) of the environmental isolates. The RAPD patterns were differentiated into 45 types (A to 2S). RAPD type A (32.3%) was the most frequently found in both clinical and environmental V cholerae non-O1 strains (34.4% and 30.0%, respectively); indicating that there was a clonal relationship between some clinical and environmental isolates whereas almost all of the environmental isolates belonged to different clones. All strains were sensitive to ciprofloxacin and norfloxacin. The environmental isolates (30%) were more resistant than the clinical ones (21.9%). Resistance to sulfamethoxazole/trimethoprim and tetracycline among the clinical isolates occurred in 9.4% (6/64) in 2007, during which period the prevalence of V cholerae O1 increased. We conclude that V. cholerae non-O1/non-O139 from the aquatic environment are potentially pathogenic and this same aquatic environment may be a source of antimicrobial resistance in V. cholerae.

  3. A case of non-O1/non-O139 Vibrio cholerae septicemia and meningitis in a neonate.

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    Hao, Yingying; Wang, Yueling; Bi, Zhenwang; Sun, Baixiu; Jin, Yan; Bai, Yuanyuan; Chen, Baoli; Shao, Chunhong; Sun, Xuerong; Lu, Zhiming

    2015-06-01

    A case of septicemia with meningitis due to non-O1/non-O139 Vibrio cholerae in a neonate is reported. The genotype and phenotype of the isolate were examined in relation to the major virulence genes. The isolate was shown to be non-toxin but cytotoxin-producing, distinguished from the dominant clone of non-O1/non-O139V. cholerae by multilocus sequence typing.

  4. A case of non-O1/non-O139 Vibrio cholerae septicemia and meningitis in a neonate

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    Yingying Hao

    2015-06-01

    Full Text Available A case of septicemia with meningitis due to non-O1/non-O139 Vibrio cholerae in a neonate is reported. The genotype and phenotype of the isolate were examined in relation to the major virulence genes. The isolate was shown to be non-toxin but cytotoxin-producing, distinguished from the dominant clone of non-O1/non-O139 V. cholerae by multilocus sequence typing.

  5. Outer membrane vesicles mediate transport of biologically active Vibrio cholerae cytolysin (VCC from V. cholerae strains.

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    Sridhar Elluri

    Full Text Available Outer membrane vesicles (OMVs released from Gram-negative bacteria can serve as vehicles for the translocation of virulence factors. Vibrio cholerae produce OMVs but their putative role in translocation of effectors involved in pathogenesis has not been well elucidated. The V. cholerae cytolysin (VCC, is a pore-forming toxin that lyses target eukaryotic cells by forming transmembrane oligomeric β-barrel channels. It is considered a potent toxin that contributes to V. cholerae pathogenesis. The mechanisms involved in the secretion and delivery of the VCC have not been extensively studied.OMVs from V. cholerae strains were isolated and purified using a differential centrifugation procedure and Optiprep centrifugation. The ultrastructure and the contents of OMVs were examined under the electron microscope and by immunoblot analyses respectively. We demonstrated that VCC from V. cholerae strain V:5/04 was secreted in association with OMVs and the release of VCC via OMVs is a common feature among V. cholerae strains. The biological activity of OMV-associated VCC was investigated using contact hemolytic assay and epithelial cell cytotoxicity test. It showed toxic activity on both red blood cells and epithelial cells. Our results indicate that the OMVs architecture might play a role in stability of VCC and thereby can enhance its biological activities in comparison with the free secreted VCC. Furthermore, we tested the role of OMV-associated VCC in host cell autophagy signalling using confocal microscopy and immunoblot analysis. We observed that OMV-associated VCC triggered an autophagy response in the target cell and our findings demonstrated for the first time that autophagy may operate as a cellular defence mechanism against an OMV-associated bacterial virulence factor.Biological assays of OMVs from the V. cholerae strain V:5/04 demonstrated that OMV-associated VCC is indeed biologically active and induces toxicity on mammalian cells and

  6. Extraintestinal Infections Caused by Non-toxigenic Vibrio cholerae non-O1/non-O139.

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    Chowdhury, Goutam; Joshi, Sangeeta; Bhattacharya, Sanjay; Sekar, Uma; Birajdar, Balaji; Bhattacharyya, Arpita; Shinoda, Sumio; Ramamurthy, Thandavarayan

    2016-01-01

    Vibrio cholerae is an aerobic, sucrose fermentative Gram-negative bacterium that generally prevails in the environment. Pathogenic V. cholerae is well-known as causative agent of acute diarrhea. Apart from enteric infections, V. cholerae may also cause other diseases. However, their role in causing extraintestinal infections is not fully known as it needs proper identification and evaluation. Four cases of extraintestinal infections due to V. cholerae non-O1/non-O139 have been investigated. The isolates were screened for phenotypic and genetic characteristics with reference to their major virulence genes. Serologically distinct isolates harbored rtx, msh, and hly but lacked enteric toxin encoding genes that are generally present in toxigenic V. cholerae. Timely detection of this organism can prevent fatalities in hospital settings. The underlying virulence potential of V. cholerae needs appropriate testing and intervention.

  7. [Studies on the enteropathogenic mechanism of non-O1 Vibrio cholerae. I. Enteropathogenicity of clinical and environmental isolates].

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    Gyobu, Y; Kodama, H; Sato, S

    1991-05-01

    Enteropathogenicity and plasmid DNA of clinical and environmental isolates of non-O1 V. cholerae were examined. Results were as follows: 1). The frequencies of enteropathogenic strains judged by the results from both ligated rabbit ileal loop (RIL) and suckling mouse tests were 36/38 (95%) for isolates from overseas travellers, 15/15 (100%) for isolates from food poisoning, 33/44 (75%) for isolates from fish and sea water, and 1/10 (10%) for isolates from river water. 2). Plasmid DNA was detected in eight of the 40 isolates examined, but the presence of plasmid did not correlate with enteropathogenicity. These results indicate that approximately three fourths of the strains isolated from fish and sea water are enteropathogenic, and that the genes controlling the enteropathogenicity of this organism probably exist in chromosomal DNA.

  8. Rapid identification of vibrio-cholerae O1 by coaglutination test using mono-specifis antibody

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    Bazargan SA

    1996-07-01

    Full Text Available In our investigation, rabbit hyper-immune serum to V.cholerae ogawa was absorbed with V.cholerae inaba whole-cells and vice versa. Applying ammonium sulphate precipitation method, mono-specific g globulins were purified and concentrated from the absorbed whole serum. These antibodies were fixed on staphylococcus cowan 1 NCTC-8325 whole-cells, using different chemical fixatives. It was observed that maximum fixation of g globulin to protein-A was achieved by 1-propanol 50% at 3 hours, which revealed through single radial immuno-diffusion techniqe. The rectal swab samples were cultured in an enrichment bile-peptons broth. After 5 hours 37°C while agitations, one drop of each sample was mixed with one drop of vibrio-cholerae bivalent mono-specific coagglutination reagent (VBCR. The results were read after 2 to 3 minutes. Finally though statistical analysis sensitivity and specificity of coagglutination test were calculated to be 95.1% and 99.2% respectively, when compared to positive & negative controls and conventional culture methods. Using VBCR, coagglutination test can be therefore considered as a simple, reliable and rapid method to detect V.cholerae O1 in the stool of patients in endemic area and less equipped laboratories

  9. Intranasal immunization with recombinant toxin-coregulated pilus and cholera toxin B subunit protects rabbits against Vibrio cholerae O1 challenge.

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    Kundu, Juthika; Mazumder, Rupa; Srivastava, Ranjana; Srivastava, Brahm S

    2009-07-01

    Intranasal immunization, a noninvasive method of vaccination, has been found to be effective in inducing systemic and mucosal immune responses. The present study was aimed at investigating the efficacy of intranasal immunization in inducing mucosal immunity in experimental cholera by subunit recombinant protein vaccines from Vibrio cholerae O1. The structural genes encoding toxin-coregulated pilus A (TcpA) and B subunit of cholera toxin (CtxB) from V. cholerae O1 were cloned and expressed in Escherichia coli. Rabbits were immunized intranasally with purified TcpA and CtxB alone or a mixture of TcpA and CtxB. Immunization with TcpA and CtxB alone conferred, respectively, 41.1% and 70.5% protection against V. cholerae challenge, whereas immunization with a mixture of both antigens conferred complete (100%) protection, as assayed in the rabbit ileal loop model. Serum titers of immunoglobulin G (IgG) antibodies to TcpA and CtxB, and anti-TcpA- and anti-CtxB-specific sIgA in intestinal lavage of vaccinated animals were found to be significantly elevated compared with unimmunized controls. Vibriocidal antibodies were detected at remarkable levels in rabbits receiving TcpA antigen and their titers correlated with protection. Thus, mucosal codelivery of pertinent cholera toxoids provides enhanced protection against experimental cholera.

  10. Genotypic and PFGE/MLVA analyses of Vibrio cholerae O1: geographical spread and temporal changes during the 2007-2010 cholera outbreaks in Thailand.

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    Kazuhisa Okada

    Full Text Available BACKGROUND: Vibrio cholerae O1 El Tor dominated the seventh cholera pandemic which occurred in the 1960s. For two decades, variants of V. cholerae O1 El Tor that produce classical cholera toxin have emerged and spread globally, replacing the prototypic El Tor biotype. This study aims to characterize V. cholerae O1 isolates from outbreaks in Thailand with special reference to genotypic variations over time. METHODS/FINDINGS: A total of 343 isolates of V. cholerae O1 from cholera outbreaks from 2007 to 2010 were investigated, and 99.4% were found to carry the classical cholera toxin B subunit (ctxB and El Tor rstR genes. Pulsed-field gel electrophoresis (PFGE differentiated the isolates into 10 distinct pulsotypes, clustered into two major groups, A and B, with an overall similarity of 88%. Ribotyping, multiple-locus variable-number tandem-repeat analysis (MLVA, and PCR to detect Vibrio seventh pandemic island II (VSP-II related genes of randomly selected isolates from each pulsotype corresponded to the results obtained by PFGE. Epidemiological investigations revealed that MLVA type 2 was strongly associated with a cholera outbreak in northeastern Thailand in 2007, while MLVA type 7 dominated the outbreaks of the southern Gulf areas in 2009 and MLVA type 4 dominated the outbreaks of the central Gulf areas during 2009-2010. Only MLVA type 16 isolates were found in a Thai-Myanmar border area in 2010, whereas those of MLVA types 26, 39, and 41 predominated this border area in 2008. Type 39 then disappeared 1-2 years later as MLVA type 41 became prevalent. Type 41 was also found to infect an outbreak area. CONCLUSIONS: MLVA provided a high-throughput genetic typing tool for understanding the in-depth epidemiology of cholera outbreaks. Our epidemiological surveys suggest that some clones of V. cholerae O1 with similar but distinctive genetic traits circulate in outbreak sites, while others disappear over time.

  11. Vibrio cholerae O1 with Reduced Susceptibility to Ciprofloxacin and Azithromycin Isolated from a Rural Coastal Area of Bangladesh

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    Rashed, Shah M.; Hasan, Nur A.; Alam, Munirul; Sadique, Abdus; Sultana, Marzia; Hoq, Md. Mozammel; Sack, R. Bradley; Colwell, Rita R.; Huq, Anwar

    2017-01-01

    Cholera outbreaks occur each year in the remote coastal areas of Bangladesh and epidemiological surveillance and routine monitoring of cholera in these areas is challenging. In this study, a total of 97 Vibrio cholerae O1 isolates from Mathbaria, Bangladesh, collected during 2010 and 2014 were analyzed for phenotypic and genotypic traits, including antimicrobial susceptibility. Of the 97 isolates, 95 possessed CTX-phage mediated genes, ctxA, ace, and zot, and two lacked the cholera toxin gene, ctxA. Also both CTX+ and CTX− V. cholerae O1 isolated in this study carried rtxC, tcpAET, and hlyA. The classical cholera toxin gene, ctxB1, was detected in 87 isolates, while eight had ctxB7. Of 95 CTX+ V. cholerae O1, 90 contained rstRET and 5 had rstRCL. All isolates, except two, contained SXT related integrase intSXT. Resistance to penicillin, streptomycin, nalidixic acid, sulfamethoxazole-trimethoprim, erythromycin, and tetracycline varied between the years of study period. Most importantly, 93% of the V. cholerae O1 were multidrug resistant. Six different resistance profiles were observed, with resistance to streptomycin, nalidixic acid, tetracycline, and sulfamethoxazole-trimethoprim predominant every year. Ciprofloxacin and azithromycin MIC were 0.003–0.75 and 0.19–2.00 μg/ml, respectively, indicating reduced susceptibility to these antibiotics. Sixteen of the V. cholerae O1 isolates showed higher MIC for azithromycin (≥0.5 μg/ml) and were further examined for 10 macrolide resistance genes, erm(A), erm(B), erm(C), ere(A), ere(B), mph(A), mph(B), mph(D), mef(A), and msr(A) with none testing positive for the macrolide resistance genes. PMID:28270803

  12. Cochleates Derived from Vibrio cholerae O1 Proteoliposomes: The Impact of Structure Transformation on Mucosal Immunisation

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    Reinaldo Acevedo; Oliver Pérez; Caridad Zayas; Pérez, José L.; Adriana Callicó; Bárbara Cedré; Luis García; David Mckee; Mullen, Alexander B.; Valerie A. Ferro

    2012-01-01

    Cochleates are phospholipid-calcium precipitates derived from the interaction of anionic lipid vesicles with divalent cations. Proteoliposomes from bacteria may also be used as a source of negatively charged components, to induce calcium-cochleate formation. In this study, proteoliposomes from V. cholerae O1 (PLc) (sized 160.7±1.6 nm) were transformed into larger (16.3±4.6 µm) cochleate-like structures (named Adjuvant Finlay Cochleate 2, AFCo2) and evaluated by electron microscopy (EM). Measu...

  13. Isolation and characterization of protective anti-LPS nanobody against V. cholerae O1 recognizing Inaba and Ogawa serotypes.

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    Ebrahimizadeh, Walead; Mousavi Gargari, Seyedlatif; Rajabibazl, Masoumeh; Safaee Ardekani, Leila; Zare, Hamed; Bakherad, Hamid

    2013-05-01

    Vibrio cholerae is considered one of the major health threats in developing countries. Lack of efficient vaccine, short incubating time of the disease, and bacterium ability to survive in aquatic environment have made cholera one of the most epidemic diseases yet known. The lipopolysaccharide is one of the bacterium key antigens used to classify V. cholerae into 206 serogroups. V. cholerae serogroup O1 is a causative agent of all cholera pandemics. Research has shown that anti-lipopolysaccharide (LPS) antibodies could provide protective immunity in cholera cases. In this research, we used N-terminal fragments of the camel's heavy-chain antibodies called VHH or nanobodies and produced a phagemid library. The obtained library was panned against V. cholerae O1 LPS, and four monoclonal nanobodies were isolated. Isolated nanobodies were tested in LPS ELISA and bacterial ELISA. The nanobody with the highest affinity toward the bacterium was used in an in vivo challenge and successfully neutralized the bacterium infection. The isolated nanobody showed high thermostability and proteolytic resistance in characterization tests.

  14. Detection of Vibrio cholerae O1 and O139 in environmental waters of rural Bangladesh: a flow-cytometry-based field trial.

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    Righetto, L; Zaman, R U; Mahmud, Z H; Bertuzzo, E; Mari, L; Casagrandi, R; Gatto, M; Islam, S; Rinaldo, A

    2015-08-01

    Presence of Vibrio cholerae serogroups O1 and O139 in the waters of the rural area of Matlab, Bangladesh, was investigated with quantitative measurements performed with a portable flow cytometer. The relevance of this work relates to the testing of a field-adapted measurement protocol that might prove useful for cholera epidemic surveillance and for validation of mathematical models. Water samples were collected from different water bodies that constitute the hydrological system of the region, a well-known endemic area for cholera. Water was retrieved from ponds, river waters, and irrigation canals during an inter-epidemic time period. Each sample was filtered and analysed with a flow cytometer for a fast determination of V. cholerae cells contained in those environments. More specifically, samples were treated with O1- and O139-specific antibodies, which allowed precise flow-cytometry-based concentration measurements. Both serogroups were present in the environmental waters with a consistent dominance of V. cholerae O1. These results extend earlier studies where V. cholerae O1 and O139 were mostly detected during times of cholera epidemics using standard culturing techniques. Furthermore, our results confirm that an important fraction of the ponds' host populations of V. cholerae are able to self-sustain even when cholera cases are scarce. Those contaminated ponds may constitute a natural reservoir for cholera endemicity in the Matlab region. Correlations of V. cholerae concentrations with environmental factors and the spatial distribution of V. cholerae populations are also discussed.

  15. Circulation of a Quorum-Sensing-Impaired Variant of Vibrio cholerae Strain C6706 Masks Important Phenotypes.

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    Stutzmann, Sandrine; Blokesch, Melanie

    2016-01-01

    Vibrio cholerae, the causative agent of cholera, is a model organism for studying virulence regulation, biofilm formation, horizontal gene transfer, and the cell-to-cell communication known as quorum sensing (QS). As in any research field, discrepancies between data from diverse laboratories are sometimes observed for V. cholerae. Such discrepancies are often caused by the use of diverse patient or environmental isolates. In this study, we investigated the inability of a few laboratories to reproduce high levels of natural transformation, a mode of horizontal gene transfer that is specifically induced on chitinous surfaces. This irreproducibility was mostly related to one specific isolate of V. cholerae: the O1 El Tor C6706 strain. C6706 was previously described as QS proficient, an important prerequisite for the induction of natural competence for transformation. To elucidate the underlying problem, we collected seven isolates of the same C6706 strain from different research laboratories in North America and Europe and compared their phenotypes. Importantly, we observed a split response with respect to QS-related gene expression, including chitin-induced natural competence and type VI secretion (T6S). While approximately half of the strains behaved as reported for several other O1 El Tor pandemic isolates that are commonly studied in the laboratory, the other half were significantly impaired in QS-related expression patterns. This impairment was caused by a mutation in a QS-related gene (luxO). We conclude that the circulation of such QS-impaired wild-type strains is responsible for masking several important phenotypes of V. cholerae, including natural competence for transformation and T6S. IMPORTANCE Phenotypic diversity between laboratory-domesticated bacterial strains is a common problem and often results in the failed reproduction of published data. However, researchers rarely compare such strains to elucidate the underlying mutation(s). In this study, we

  16. The first case of bacteraemia due to non-O1/non-O139 Vibrio cholerae in a type 2 diabetes mellitus patient in mainland China.

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    Lu, Binghuai; Zhou, Haijian; Li, Dong; Li, Fengjuan; Zhu, Fengxia; Cui, Yanchao; Huang, Lei; Wang, Duochun

    2014-08-01

    Bacteraemia due to non-O1/non-O139 Vibrio cholerae is rarely documented in mainland China. We report such a case in a 70-year-old male with type 2 diabetes mellitus. The clinical features, phenotypic analyses, and presence of a panel of known virulence genes in the isolated strain are described. To the best of our knowledge, this is the first reported case of bacteraemia due to this strain in a T2DM patient without other coexisting underlying diseases in mainland China.

  17. Occurrence of virulence genes among Vibrio cholerae and Vibrio parahaemolyticus strains from treated wastewaters.

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    Khouadja, Sadok; Suffredini, Elisabetta; Baccouche, Besma; Croci, Luciana; Bakhrouf, Amina

    2014-10-01

    Pathogenic Vibrio species are an important cause of foodborne illnesses. The aim of this study was to describe the occurrence of potentially pathogenic Vibrio species in the final effluents of a wastewater treatment plant and the risk that they may pose to public health. During the 1-year monitoring, a total of 43 Vibrio strains were isolated: 23 Vibrio alginolyticus, 1 Vibrio cholerae, 4 Vibrio vulnificus, and 15 Vibrio parahaemolyticus. The PCR investigation of V. parahaemolyticus and V. cholerae virulence genes (tlh, trh, tdh, toxR, toxS, toxRS, toxT, zot, ctxAB, tcp, ace, vpi, nanH) revealed the presence of some of these genes in a significant number of strains. Intraspecies variability and genetic relationships among the environmental isolates were analyzed by random amplified polymorphic DNA-PCR (RAPD-PCR). We report the results of the first isolation and characterization of an environmental V. cholerae non-O1 non-O139 and of a toxigenic V. parahaemolyticus strain in Tunisia. We suggest that non-pathogenic Vibrio might represent a marine reservoir of virulence genes that can be transmitted between strains by horizontal transfer.

  18. Distribution of virulence-associated genes and genetic relationships in non-O1/O139 Vibrio cholerae aquatic isolates from China.

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    Li, Fengjuan; Du, Pengcheng; Li, Baisheng; Ke, Changwen; Chen, Aiping; Chen, Jie; Zhou, Haijian; Li, Jie; Morris, J Glenn; Kan, Biao; Wang, Duochun

    2014-08-01

    Non-O1/O139 Vibrio cholerae is naturally present in aquatic ecosystems and has been linked with cholera-like diarrhea and local outbreaks. The distribution of virulence-associated genes and genetic relationships among aquatic isolates from China are largely unknown. In this study, 295 aquatic isolates of V. cholerae non-O1/O139 serogroups from different regions in China were investigated. Only one isolate was positive for ctxB and harbored a rare genotype; 10 (3.4%) isolates carried several types of rstR sequences, eight of which carried rare types of toxin-coregulated pili (tcpA). Furthermore, 16 (5.4%) isolates carried incomplete (with partial open reading frames [ORFs]) vibrio seventh pandemic island I (VSP-I) or VSP-II clusters, which were further classified as 11 novel types. PCR-based analyses revealed remarkable variations in the distribution of putative virulence genes, including mshA (95.6%), hlyA (95.3%), rtxC (89.8%), rtxA (82.7%), IS1004 (52.9%), chxA (30.2%), SXT (15.3%), type III secretion system (18.0%), and NAG-ST (3.7%) genes. There was no correlation between the prevalence of putative virulence genes and that of CTX prophage or TCP genes, whereas there were correlations among the putative virulence genes. Further multilocus sequence typing (MLST) placed selected isolates (n = 70) into 69 unique sequence types (STs), which were different from those of the toxigenic O1 and O139 counterparts, and each isolate occupied a different position in the MLST tree. The V. cholerae non-O1/O139 aquatic isolates predominant in China have high genotypic diversity; these strains constitute a reservoir of potential virulence genes, which may contribute to evolution of pathogenic isolates.

  19. Necrotizing fasciitis due to Vibrio cholerae non-O1/non-O139 after exposure to Austrian bathing sites.

    Science.gov (United States)

    Hirk, Sonja; Huhulescu, Steliana; Allerberger, Franz; Lepuschitz, Sarah; Rehak, Sonja; Weil, Sandra; Gschwandtner, Elisabeth; Hermann, Michael; Neuhold, Stephanie; Zoufaly, Alexander; Indra, Alexander

    2016-02-01

    We report on two cases of necrotizing fasciitis of the lower leg due to nontoxigenic Vibrio cholerae (V. cholerae). A 73-year-old woman (case 1) and an 80-year-old man (case 2) were hospitalized with symptoms of necrotizing fasciitis on July 18 and August 15, 2015, respectively. In both cases, symptoms started the day after swimming in local ponds. Swabs gained intraoperatively and a blood culture from the male patient, yielded V. cholerae non-O1/non-O139, negative for cholera toxin gene ctx and positive for hemolysin genes hlyA and hlyB. Water samples taken from pond A on August 17, 2015 (32 days after exposure of case 1) and from pond B on August 20, 2015 (7 days after exposure of case 2) yielded non-O1/non-O139 V. cholerae in most-probable numbers of > 11,000 per 100 ml each. The occurrence of two cases of necrotizing fasciitis within a 1 month period related to two Austrian non-saline bathing waters, previously not known to harbor V. cholerae, is probably linked to the prevailing extreme weather conditions (heat wave, drought) this summer in Austria. While case 1 was discharged in good clinical condition after 73 days, case 2 died after four months of hospitalization. Public health authorities are challenged to assess the effects of long-term climate change on pathogen growth and survival in continental bodies of fresh water.

  20. Genome-wide study of the defective sucrose fermenter strain of Vibrio cholerae from the Latin American cholera epidemic.

    NARCIS (Netherlands)

    Garza, D.R.; Thompson, C.C.; Loureiro, E.C.; Dutilh, B.E.; Inada, D.T.; Junior, E.C.; Cardoso, J.F.; Nunes, M.R.; Lima, C.P. de; Silvestre, R.V.; Nunes, K.N.; Santos, E.C.; Edwards, R.A.; Vicente, A.C.; Sá Morais, L.L. de

    2012-01-01

    The 7th cholera pandemic reached Latin America in 1991, spreading from Peru to virtually all Latin American countries. During the late epidemic period, a strain that failed to ferment sucrose dominated cholera outbreaks in the Northern Brazilian Amazon region. In order to understand the genomic char

  1. Tipificación Molecular del Vibrio cholerae O1 en el Perú

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    Huguet T José

    2000-01-01

    Full Text Available Este estudio de ribotipificación en 75 cepas de Vibrio cholerae O1 permitió identificar tres variantes ribotípicas, referidas como Per1, Per2 y Per3, aisladas durante el periodo 1991- 1999 en el Perú. La variante Per1 fue reportada tanto en la etapa epidémica y endémica del cólera, mientras que Per2 y Per3 se relacionaron sólo con la etapa endémica. Los resultados mostraron además una aparición constante y mayoritaria de la variante Per1, poniendo en evidencia la emergencia de un mismo grupo clonal en los brotes epidémicos del Perú. Las variantes ribotípicas encontradas fueron comparadas con los ribotipos de diferentes cepas referenciales de V. cholerae previamente caracterizadas. Se observó una identidad total del ribotipo Per1 con la variante ribotípica de aislamientos Asiáticos (Tailandia, encontrándose además altos índices de similitud entre los ribotipos Per1, Per2 y Per3, y evidenciándose una estrecha relación entre las cepas peruanas y los aislamientos asiáticos.

  2. Inhibition of the sodium-translocating NADH-ubiquinone oxidoreductase [Na+-NQR] decreases cholera toxin production in Vibrio cholerae O1 at the late exponential growth phase.

    Science.gov (United States)

    Minato, Yusuke; Fassio, Sara R; Reddekopp, Rylan L; Häse, Claudia C

    2014-01-01

    Two virulence factors produced by Vibrio cholerae, cholera toxin (CT) and toxin-corregulated pilus (TCP), are indispensable for cholera infection. ToxT is the central regulatory protein involved in activation of CT and TCP expression. We previously reported that lack of a respiration-linked sodium-translocating NADH-ubiquinone oxidoreductase (Na(+)-NQR) significantly increases toxT transcription. In this study, we further characterized this link and found that Na(+)-NQR affects toxT expression only at the early-log growth phase, whereas lack of Na(+)-NQR decreases CT production after the mid-log growth phase. Such decreased CT production was independent of toxT and ctxB transcription. Supplementing a respiratory substrate, l-lactate, into the growth media restored CT production in the nqrA-F mutant, suggesting that decreased CT production in the Na(+)-NQR mutant is dependent on electron transport chain (ETC) activity. This notion was supported by the observations that two chemical inhibitors, a Na(+)-NQR specific inhibitor 2-n-Heptyl-4-hydroxyquinoline N-oxide (HQNO) and a succinate dehydrogenase (SDH) inhibitor, thenoyltrifluoroacetone (TTFA), strongly inhibited CT production in both classical and El Tor biotype strains of V. cholerae. Accordingly, we propose the main respiratory enzyme of V. cholerae, as a potential drug target to treat cholera because human mitochondria do not contain Na(+)-NQR orthologs.

  3. Coagglutination test for detecting V. Cholerae serotype O1%协同凝集试验检测O1群霍乱弧菌

    Institute of Scientific and Technical Information of China (English)

    陈兆鹏; 袁应华; 刘忠明

    2001-01-01

    Objective To develop a simple and rapid method for detecting V. cholerae O1. Methods Staphylococcus aureus Cowan I was coated with anti-V. cholerae O1 monoclonal antibodies and a coagglutination test(COA) detecting V. cholerae O1 in 6-h fecal enrichment cultures was tested. Results The detect limit of COA was 4×106 CFU/ml, and detect time was 5 min. When 92 samples were tested, the diagnostic specificity, sensitivity and accuracy of the test, compared with standard culture methods, were 100%、95.5%、97.8%,respectively. Conclusion The COA is simple and less time-consuming, and can be used as a new method for rapid detection of V. Cholerae.%目的建立一种快速简便的O1群霍乱弧菌检测方法。方法将抗O1群霍乱弧菌多价单克隆抗体包被到金黄色葡萄球菌Cowan I株上制成SPA菌体试剂,建立起协同凝集试验(COA),检测粪便标本6 h培养物中的霍乱弧菌。结果 COA检测的敏感性为4×106 CFU/ml,检测过程为5 min。92份临床标本的测定结果与培养法相比较,COA的特异性、敏感性、诊断符合率分别是100%、95.5%、97.8%。结论 COA简便、快速,可作为一种新的霍乱弧菌快速诊断方法。

  4. Characters of homogentisate oxygenase gene mutation and high clonality of the natural pigment-producing Vibrio cholerae strains

    Directory of Open Access Journals (Sweden)

    Diao Baowei

    2011-05-01

    Full Text Available Abstract Background Some microorganisms can produce pigments such as melanin, which has been associated with virulence in the host and with a survival advantage in the environment. In Vibrio cholerae, studies have shown that pigment-producing mutants are more virulent than the parental strain in terms of increased UV resistance, production of major virulence factors, and colonization. To date, almost all of the pigmented V. cholerae strains investigated have been induced by chemicals, culture stress, or transposon mutagenesis. However, during our cholera surveillance, some nontoxigenic serogroup O139 strains and one toxigenic O1 strain, which can produce pigment steadily under the commonly used experimental growth conditions, were obtained in different years and from different areas. The genes VC1344 to VC1347, which correspond to the El Tor strain N16961 genome and which comprise an operon in the tyrosine catabolic pathway, have been confirmed to be associated with a pigmented phenotype. In the present study, we investigated the mechanism of pigment production in these strains. Results Sequencing of the VC1344, VC1345, VC1346, and VC1347 genes in these pigmented strains suggested that a deletion mutation in the homogentisate oxygenase gene (VC1345 may be associated with the pigmented phenotype, and gene complementation confirmed the role of this gene in pigment production. An identical 15-bp deletion was found in the VC1345 gene of all six O139 pigment-producing strains examined, and a 10-bp deletion was found in the VC1345 gene of the O1 strain. Strict sequence conservation in the VC1344 gene but higher variance in the other three genes of this operon were observed, indicating the different stress response functions of these genes in environmental adaption and selection. On the basis of pulsed-field gel electrophoresis typing, the pigment-producing O139 strains showed high clonality, even though they were isolated in different years and from

  5. Long-term comparison of antibiotic resistance in Vibrio cholerae O1 and Shigella species between urban and rural Bangladesh.

    Science.gov (United States)

    Klontz, Erik H; Das, Sumon Kumar; Ahmed, Dilruba; Ahmed, Shahnawaz; Chisti, Mohammod Jobayer; Malek, Mohammad Abdul; Faruque, Abu Syed Golam; Klontz, Karl C

    2014-05-01

    From 2000 to 2012, Vibrio cholerae O1 and Shigella species isolates from urban Dhaka and rural Matlab were tested for resistance to all clinically relevant antibiotics in Bangladesh. Resistances in urban and rural Bangladesh tended to rise and fall together, especially a few years after the introduction of new resistance.

  6. Intranasal administration of proteoliposome-derived cochleates from Vibrio cholerae O1 induce mucosal and systemic immune responses in mice.

    Science.gov (United States)

    Acevedo, Reinaldo; Callicó, Adriana; del Campo, Judith; González, Elizabeth; Cedré, Bárbara; González, Lissette; Romeu, Belkis; Zayas, Caridad; Lastre, Miriam; Fernández, Sonsire; Oliva, Reynaldo; García, Luis; Pérez, José Luis; Pérez, Oliver

    2009-12-01

    Conservative estimates place the death toll from cholera at more than 100,000 persons each year. A particulate mucosal vaccine strategy combining antigens and immune stimulator molecules from Vibrio cholerae to overcome this problem is described. Proteoliposomes extracted from V. cholerae O1 were transformed into cochleates (AFCo2, Adjuvant Finlay cochleate 2) through a calcium inducible rotary dialysis method. Light microscopy was carried out and tubules of 16.25+/-4.57 microm in length were observed. Western blots were performed to verify the immunochemical properties of the main AFCo2 incorporated antigens, revealing full recognition of the outer membrane protein U (OmpU), lipopolysaccharide (LPS), and mannose-sensitive hemagglutinin (MSHA) antigens. AFCo2 were administered by the intranasal route using a two or three dose schedule and the immune response against V. cholerae antigens was assessed. Three AFCo2 doses were required to induce significant (p<0.05), antigen specific IgA in saliva (1.34+/-0.135) and feces (0.60+/-0.089). While, two or three doses of AFCo2 or proteoliposomes induce similar specific IgG and vibriocidal activity responses in sera. These results show for the first time that AFCo2 can be obtained from V. cholerae O1 proteoliposomes and have the potential to protect against the pathogen when administered intranasally.

  7. Retrospective Analysis of Serotype Switching of Vibrio cholerae O1 in a Cholera Endemic Region Shows It Is a Non-random Process

    Science.gov (United States)

    Karlsson, Stefan L.; Thomson, Nicholas; Mutreja, Ankur; Connor, Thomas; Sur, Dipika; Ali, Mohammad; Clemens, John; Dougan, Gordon; Holmgren, Jan; Lebens, Michael

    2016-01-01

    Genomic data generated from clinical Vibrio cholerae O1 isolates collected over a five year period in an area of Kolkata, India with seasonal cholera outbreaks allowed a detailed genetic analysis of serotype switching that occurred from Ogawa to Inaba and back to Ogawa. The change from Ogawa to Inaba resulted from mutational disruption of the methyltransferase encoded by the wbeT gene. Re-emergence of the Ogawa serotype was found to result either from expansion of an already existing Ogawa clade or reversion of the mutation in an Inaba clade. Our data suggests that such transitions are not random events but rather driven by as yet unidentified selection mechanisms based on differences in the structure of the O1 antigen or in the serotype-determining wbeT gene. PMID:27706170

  8. Retrospective Analysis of Serotype Switching of Vibrio cholerae O1 in a Cholera Endemic Region Shows It Is a Non-random Process.

    Science.gov (United States)

    Karlsson, Stefan L; Thomson, Nicholas; Mutreja, Ankur; Connor, Thomas; Sur, Dipika; Ali, Mohammad; Clemens, John; Dougan, Gordon; Holmgren, Jan; Lebens, Michael

    2016-10-01

    Genomic data generated from clinical Vibrio cholerae O1 isolates collected over a five year period in an area of Kolkata, India with seasonal cholera outbreaks allowed a detailed genetic analysis of serotype switching that occurred from Ogawa to Inaba and back to Ogawa. The change from Ogawa to Inaba resulted from mutational disruption of the methyltransferase encoded by the wbeT gene. Re-emergence of the Ogawa serotype was found to result either from expansion of an already existing Ogawa clade or reversion of the mutation in an Inaba clade. Our data suggests that such transitions are not random events but rather driven by as yet unidentified selection mechanisms based on differences in the structure of the O1 antigen or in the serotype-determining wbeT gene.

  9. Isolation and identification of an atypical non-O1 Vibrio cholerae%1株不典型非O1群霍乱弧菌的发现及鉴定

    Institute of Scientific and Technical Information of China (English)

    吴至成; 覃西; 伍丽娴; 翟英超; 王小娟; 李影林

    2012-01-01

    OBJECTIVE To identify a strain of pathogenic bacterium isolated from the blood of a cirrhotic patient. METHODS The phenotypic characteristics (including morphology, culture characteristics, and biochemical characteristics) and the serologic characteristics were identified according to the standard analytical protocol for Vibrio cholerae, and the 16S rDNA sequence of the isolate was analyzed. RESULTS There was one strain of pathogen (strain HN 9837) which was isolated from the blood of the patient and resembled V. cholerae in its morphological, cultural, biochemical, and serological characters; remarkably, it showed atypical biochemical characteristics such as negative results by tests for mannitol fermentation, lysine decarboxylase and citrate utilization. CONCLUSION It is the first report in Hainan province that non-O1 V. cholerae is isolated from the blood of a cirrhotic patient. The isolate with the following atypical reactions was shown to be V. cholerae: mannitol negative, lysine decarboxylase negative and citrate negative.%目的 对1株从肝硬化患者血液中分离的病原菌进行鉴定.方法 按霍乱弧菌的鉴定标准和方法进行细菌学表型特征(包括形态学、培养特性、生化特征)、血清学鉴定及16S rDNA序列分析.结果 从患者血液中分离获得1株病原菌HN 9837,该菌株的形态学、培养特性、生化特征、血清学诊断和分子生物学诊断结果基本符合非O1群霍乱弧菌定义;但与普通霍乱弧菌在甘露醇发酵及赖氨酸利用、柠檬酸盐利用试验结果存在差异.结论 海南省首次在肝硬化患者血液中分离出非O1群霍乱弧菌,该菌为1株不发酵甘露醇、赖氨酸且柠檬酸盐利用试验阴性的不典型霍乱弧菌.

  10. Emergence of Tetracycline Resistant Vibrio cholerae O1 Biotype El Tor Serotype Ogawa with Classical ctxB Gene from a Cholera Outbreak in Odisha, Eastern India.

    Science.gov (United States)

    Jain, M; Kumar, P; Goel, A K

    2016-01-01

    In September 2010, a cholera outbreak was reported from Odisha, Eastern India. V. cholerae isolated from the clinical samples were biochemically and serologically confirmed as serogroup O1, biotype El Tor, and serotype Ogawa. Multiplex PCR screening revealed the presence of various genes, namely, ompW, ctxB, zot, rfbO1, tcp, ace, hlyA, ompU, rtx, and toxR, in all of the isolates. The isolates were resistant to co-trimoxazole, nalidixic acid, polymyxin B, spectinomycin, streptomycin, sulfamethoxazole, tetracycline, trimethoprim, and vibriostatic agent 2,4-diamino-6,7-diisopropylpteridine (O/129). Minimum inhibitory concentration of tetracycline decreased in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP), suggesting the involvement of efflux pumps. PCR analysis confirmed the presence of class I integrons as well as SXT elements harbouring antibiotic resistance genes in all isolates. Sequencing revealed the presence of ctxB gene of classical biotype in all the isolates. The isolates harboured an RS1-CTX prophage array with El Tor type rstR and classical ctxB on the large chromosome. The study indicated that the V. cholerae El Tor variants are evolving in the area with better antibiotic resistance and virulence potential.

  11. Distribution and molecular characteristics of Vibrio cholerae O1 El Tor isolates recovered in Guangdong Province, China, 1961-2013.

    Science.gov (United States)

    Li, Baisheng; Chen, Rongfeng; Wang, Duochun; Tan, Hailing; Ke, Bixia; He, Dongmei; Ke, Changwen; Zhang, Yonghui

    2016-01-01

    China's Guangdong Province is located along the same latitude as Kolkata, India and Dhaka, Bangladesh, and is also considered a source of epidemic cholera. However, molecular description and the genetic relationships between Vibrio cholerae O1 El Tor isolates in Guangdong remain unclear. In this study, 381 clinical V. cholerae O1 isolates recovered from cholera cases presenting in Guangdong between 1961 and 2013 were investigated by PCR, amplicon sequencing and pulsed-field gel electrophoresis (PFGE). During this time frame, four distinct epidemic periods (1-4) were observed based on the different dominant serotype leading its epidemic, correspond to years; or time periods from/to 1961-1969, 1978-1989, 1990-2000, 2001-2013, respectively. Molecular analysis of representative isolates indicated that a single dominating clone was associated with each epidemic stage. All isolates from periods 1 and 2 carried the typical El Tor ctxB; this allele was displaced by classical ctxB beginning in 1993. However all isolates carried the El Tor-specific toxin-coregulated pili subunit A (tcpA). Isolates were grouped into five clusters on the basis of Not I enzyme digested PFGE, and the first four clusters were associated with specific periods, cluster I (period 1), II (period 3), III (period 2) and IV (period 4), respectively. While cluster V consisted of isolates from all four epidemic periods, but was most heterogeneous in appearance. Our data indicate genetic variations that shape the relationship among emerging isolates of V. cholerae O1 in Guangdong Province contribute to the 7th global pandemic.

  12. Cochleates derived from Vibrio cholerae O1 proteoliposomes: the impact of structure transformation on mucosal immunisation.

    Science.gov (United States)

    Acevedo, Reinaldo; Pérez, Oliver; Zayas, Caridad; Pérez, José L; Callicó, Adriana; Cedré, Bárbara; García, Luis; Mckee, David; Mullen, Alexander B; Ferro, Valerie A

    2012-01-01

    Cochleates are phospholipid-calcium precipitates derived from the interaction of anionic lipid vesicles with divalent cations. Proteoliposomes from bacteria may also be used as a source of negatively charged components, to induce calcium-cochleate formation. In this study, proteoliposomes from V. cholerae O1 (PLc) (sized 160.7±1.6 nm) were transformed into larger (16.3±4.6 µm) cochleate-like structures (named Adjuvant Finlay Cochleate 2, AFCo2) and evaluated by electron microscopy (EM). Measurements from transmission EM (TEM) showed the structures had a similar size to that previously reported using light microscopy, while observations from scanning electron microscopy (SEM) indicated that the structures were multilayered and of cochleate-like formation. The edges of the AFCo2 structures appeared to have spaces that allowed penetration of negative stain or Ovalbumin labeled with Texas Red (OVA-TR) observed by epi-fluorescence microscopy. In addition, freeze fracture electron microscopy confirmed that the AFCo2 structures consisted of multiple overlapping layers, which corresponds to previous descriptions of cochleates. TEM also showed that small vesicles co-existed with the larger cochleate structures, and in vitro treatment with a calcium chelator caused the AFCo2 to unfold and reassemble into small proteoliposome-like structures. Using OVA as a model antigen, we demonstrated the potential loading capacity of a heterologous antigen and in vivo studies showed that with simple admixing and administration via intragastric and intranasal routes AFCo2 provided enhanced adjuvant properties compared with PLc.

  13. Characterization of Vibrio cholerae O1 ElTor typing phage S5.

    Science.gov (United States)

    Mitra, K; Ghosh, A N

    2007-01-01

    S5 (ATCC No. 51352-B2), a Vibrio cholerae O1 ElTor typing phage was characterized. The growth characteristics and inactivation kinetics (thermal, UV and pH) of this lytic phage were investigated. Phage morphology was examined by electron microscopy and was classified as belonging to the family Podoviridae. The S5 phage genome is shown to be a linear double-stranded 39-kb-long DNA as determined by electron microscopy and restriction digestion. Partial denaturation maps were constructed and were used to show that the DNA is non-permuted and terminally redundant. The replication origin of this T7-like phage was visualized by electron microscopy. The polarity of packaging of S5 DNA in the phage head was determined. SDS-PAGE of phage S5 shows two major structural polypeptides of 50 and 42 kDa. A 3D structure of the phage head was reconstructed at a resolution of 37 A using Cryo-EM and a single-particle reconstruction technique.

  14. Viabilidade de Vibrio cholerae O1 em diferentes tipos de águas em condições experimentais

    Directory of Open Access Journals (Sweden)

    Nogueira Joseli Maria da Rocha

    2002-01-01

    Full Text Available A natureza endêmica e sazonal da cólera depende da sobrevivência de Vibrio cholerae O1 em estado viável, mas não necessariamente cultivável em nichos ambientais aquáticos durante períodos interepidêmicos, sendo de suma importância o estudo da sobrevivência deste microrganismo nesses locais. Para tal, foram coletadas, semanalmente, alíquotas de água pertencentes a duas lagoas e dois rios do Estado do Rio de Janeiro. Esses volumes foram divididos em duas porções idênticas, uma das quais foi autoclavada. Uma diluição padronizada de V. cholerae O1 Inaba e de V. cholerae O1 Ogawa, foi inoculada em três alíquotas de 100ml dessas diferentes águas e mantidas em diferentes temperaturas. A sobrevivência desses microorganismos no âmbito aquático sob esses diferentes fatores foi então analisada. Os resultados demonstraram que o V. cholerae sorogrupo O1, independente do sorotipo, é capaz de se manter em água com salinidade abaixo de 0,5? e em diferentes temperaturas, por períodos suficientes para sua disseminação através de "corpos d'água", demonstrando a necessidade de monitoramento constante em áreas de possível contaminação, principalmente onde a água é utilizada para o consumo, evitando assim, a disseminação da doença para as populações próximas a esses ambientes.

  15. Characterization of pathogenicity island prophage in clinical and environmental strains of Vibrio cholerae.

    Science.gov (United States)

    Mohammadi-Barzelighi, H; Bakhshi, B; Rastegar Lari, A; Pourshafie, M R

    2011-12-01

    In this study 86 isolates of Vibrio cholerae were analysed for their adhesive properties and the presence of pathogenicity island genes. With the exception of three isolates, all of the other clinical isolates (92.5%) contained an intact TCP (toxin-co-regulated pilus) gene cluster. In contrast, 95% of all environmental non-O1-non-O139 isolates were negative for the TCP gene cluster. The majority of clinical isolates (82.5%) possessed the complete vibrio pathogenicity island (VPI) gene cluster and had a similar RFLP pattern, while only a single environmental strain possessed an almost complete VPI cluster (lacking 0.4 kb in the tcpA and toxT region). The result showed that the isolates with tcpA(+)/toxT(+) had a strong attachment for HT-29 and Vero cells, whereas isolates with tcpA(+)/toxT(-) or tcpA(-)/toxT(-) genomic characteristics showed no autoagglutination and weak attachment for the cell lines. Two environmental strains (tcpA(-)/toxT(-)) showed strong adhesive properties to the cell lines, indicating that non-fimbrial adhesive factors are involved in the environmental V. cholerae strains in the absence of TCP.

  16. Non-O1/non-O139 Vibrio cholerae septicaemia in a Saudi man: a case report

    Science.gov (United States)

    El-Hossary, Dalia; Jiman-Fatani, Asif; Al-Ghamdi, Rahaf

    2017-01-01

    Background. The non-O1/non-O139 serogroups of Vibrio cholerae occur in diverse natural niches, and usually cause mild and self-limiting gastrointestinal illness. However, they have well-documented potential to cause invasive and extra-intestinal infections among immunocompromised patients. Furthermore, their ability to grow in low-salinity surface water, and the existence of asymptomatic human carriers, suggest novel acquisition routes for this unusual infection, even in people without obvious risk factors. Case presentation. A 62-year-old man presented with epigastric pain, vomiting and fever. The patient had a history of diabetes and cholecystectomy, although our initial examination did not reveal any significant findings that might indicate V. cholerae infection. However, blood cultures subsequently revealed the presence of V. cholerae, which was positively identified using both conventional and modern non-conventional technologies. The identity of the V. cholerae isolate was confirmed using Vitek MS (matrix–assisted laser desorption ionization-time of flight MS) and the FilmArray system, in addition to its initial identification using the Vitek 2 system. The septicaemia was successfully treated using a 14 day course of ciprofloxacin. Conclusion. The present case highlights the need to remain highly suspicious of non-O1/non-O139 V. cholerae infections in patients with known risk factors, as well as in healthy individuals with epidemiological exposure and compatible clinical symptoms. Special care should be taken to avoid false-positive results from confirmatory laboratory tests, as the organism can grow in fresh water, and the results should be verified using multiple methods. PMID:28348803

  17. Genome-wide study of the defective sucrose fermenter strain of Vibrio cholerae from the Latin American cholera epidemic.

    Directory of Open Access Journals (Sweden)

    Daniel Rios Garza

    Full Text Available The 7th cholera pandemic reached Latin America in 1991, spreading from Peru to virtually all Latin American countries. During the late epidemic period, a strain that failed to ferment sucrose dominated cholera outbreaks in the Northern Brazilian Amazon region. In order to understand the genomic characteristics and the determinants of this altered sucrose fermenting phenotype, the genome of the strain IEC224 was sequenced. This paper reports a broad genomic study of this strain, showing its correlation with the major epidemic lineage. The potentially mobile genomic regions are shown to possess GC content deviation, and harbor the main V. cholera virulence genes. A novel bioinformatic approach was applied in order to identify the putative functions of hypothetical proteins, and was compared with the automatic annotation by RAST. The genome of a large bacteriophage was found to be integrated to the IEC224's alanine aminopeptidase gene. The presence of this phage is shown to be a common characteristic of the El Tor strains from the Latin American epidemic, as well as its putative ancestor from Angola. The defective sucrose fermenting phenotype is shown to be due to a single nucleotide insertion in the V. cholerae sucrose-specific transportation gene. This frame-shift mutation truncated a membrane protein, altering its structural pore-like conformation. Further, the identification of a common bacteriophage reinforces both the monophyletic and African-Origin hypotheses for the main causative agent of the 1991 Latin America cholera epidemics.

  18. The Lake Chad Basin, an Isolated and Persistent Reservoir of Vibrio cholerae O1: A Genomic Insight into the Outbreak in Cameroon, 2010

    Science.gov (United States)

    Kaas, Rolf S.; Ngandjio, Antoinette; Nzouankeu, Ariane; Siriphap, Achiraya; Fonkoua, Marie-Christine; Aarestrup, Frank M.; Hendriksen, Rene S.

    2016-01-01

    The prevalence of reported cholera was relatively low around the Lake Chad basin until 1991. Since then, cholera outbreaks have been reported every couple of years. The objective of this study was to investigate the 2010/2011 Vibrio cholerae outbreak in Cameroon to gain insight into the genomic make-up of the V. cholerae strains responsible for the outbreak. Twenty-four strains were isolated and whole genome sequenced. Known virulence genes, resistance genes and integrating conjugative element (ICE) elements were identified and annotated. A global phylogeny (378 genomes) was inferred using a single nucleotide polymorphism (SNP) analysis. The Cameroon outbreak was found to be clonal and clustered distant from the other African strains. In addition, a subset of the strains contained a deletion that was found in the ICE element causing less resistance. These results suggest that V. cholerae is endemic in the Lake Chad basin and different from other African strains. PMID:27191718

  19. Epidemiologic and Drug Resistance Pattern of Vibrio cholerae O1 Biotype El Tor, Serotype Ogawa, in the 2011 Cholera Outbreak, in Alborz Province, Iran

    Science.gov (United States)

    Barati, Hojatolah; Moradi, Ghobad; Rasouli, Mohammad Aziz; Mohammadi, Parvin

    2015-01-01

    Background: Although the national guidelines recommend special antibiotics, based on the antibiogram of National Reference Laboratory, it seems that, because of uncontrolled usage of antibiotics in the society and due to the changes in the serotypes causing the disease, it is essential to monitor the status of drug resistance, permanently, and to revise the current prescriptions guidelines. Objectives: This study aimed to assess the epidemiological aspects and drug resistance pattern of Vibrio cholerae O1, biotype El Tor, serotype Ogawa, in cholera outbreak, in Alborz province in Iran, during 2011. Materials and Methods: This is a cross-sectional study, which reviews a cholera epidemic that occurred in Iran. A total of 9844 specimens were taken from suspected cases, among diarrheal patients, via rectal swabs. The specimens were placed in Cary-Blair transport medium and sent to laboratory. Samples were enriched, in alkaline peptone water, and isolated on thiosulphate-citrate-bile salt-sucrose agar. From the 244 confirmed cases, 239 cases underwent antibiogram test, via disk diffusion method and based on national committee for clinical laboratory standards (NCCLS) instructions. The standard Escherichia coli ATCC 25922 was used for antibiogram quality control and, eventually, all results were interpreted and reported using NCCLS standard table. Results: In total, until October 22, 2011, which was announced as the end of outbreak, 9844 samples were taken from diarrheal patients. Regarding the type of V. cholerae, 244 El Tor biotype positive cases were reported. The case fatality rate was 1.3%. The mean age of patients was 37.8 years and the highest incidence rate occurred in the age group 21 - 30 years. After conducting antibiotic susceptibility test in the 244 V. cholerae, biotype El Tor, serotype Ogawa, it was found that ciprofloxacin had the highest level of antibiotic susceptibility (99.6%) and the highest level of antibiotic resistance was observed in co

  20. Cochleates derived from Vibrio cholerae O1 proteoliposomes: the impact of structure transformation on mucosal immunisation.

    Directory of Open Access Journals (Sweden)

    Reinaldo Acevedo

    Full Text Available Cochleates are phospholipid-calcium precipitates derived from the interaction of anionic lipid vesicles with divalent cations. Proteoliposomes from bacteria may also be used as a source of negatively charged components, to induce calcium-cochleate formation. In this study, proteoliposomes from V. cholerae O1 (PLc (sized 160.7±1.6 nm were transformed into larger (16.3±4.6 µm cochleate-like structures (named Adjuvant Finlay Cochleate 2, AFCo2 and evaluated by electron microscopy (EM. Measurements from transmission EM (TEM showed the structures had a similar size to that previously reported using light microscopy, while observations from scanning electron microscopy (SEM indicated that the structures were multilayered and of cochleate-like formation. The edges of the AFCo2 structures appeared to have spaces that allowed penetration of negative stain or Ovalbumin labeled with Texas Red (OVA-TR observed by epi-fluorescence microscopy. In addition, freeze fracture electron microscopy confirmed that the AFCo2 structures consisted of multiple overlapping layers, which corresponds to previous descriptions of cochleates. TEM also showed that small vesicles co-existed with the larger cochleate structures, and in vitro treatment with a calcium chelator caused the AFCo2 to unfold and reassemble into small proteoliposome-like structures. Using OVA as a model antigen, we demonstrated the potential loading capacity of a heterologous antigen and in vivo studies showed that with simple admixing and administration via intragastric and intranasal routes AFCo2 provided enhanced adjuvant properties compared with PLc.

  1. Cholera.

    Science.gov (United States)

    Lippi, Donatella; Gotuzzo, Eduardo; Caini, Saverio

    2016-08-01

    Cholera is an acute disease of the gastrointestinal tract caused by Vibrio cholerae. Cholera was localized in Asia until 1817, when a first pandemic spread from India to several other regions of the world. After this appearance, six additional major pandemics occurred during the 19th and 20th centuries, the latest of which originated in Indonesia in the 1960s and is still ongoing. In 1854, a cholera outbreak in Soho, London, was investigated by the English physician John Snow (1813 to 1858). He described the time course of the outbreak, managed to understand its routes of transmission, and suggested effective measures to stop its spread, giving rise to modern infectious disease epidemiology. The germ responsible for cholera was discovered twice: first by the Italian physician Filippo Pacini during an outbreak in Florence, Italy, in 1854, and then independently by Robert Koch in India in 1883, thus favoring the germ theory over the miasma theory of disease. Unlike many other infectious diseases, such as plague, smallpox, and poliomyelitis, cholera persists as a huge public health problem worldwide, even though there are effective methods for its prevention and treatment. The main reasons for its persistence are socioeconomic rather than purely biological; cholera flourishes where there are unsatisfactory hygienic conditions and where a breakdown of already fragile sanitation and health infrastructure occurs because of natural disasters or humanitarian crises.

  2. The Lake Chad Basin, an Isolated and Persistent Reservoir of Vibrio cholerae O1: A Genomic Insight into the Outbreak in Cameroon, 2010

    DEFF Research Database (Denmark)

    Kaas, Rolf Sommer; Ngandjio, Antoinette; Nzouankeu, Ariane

    2016-01-01

    The prevalence of reported cholera was relatively low around the Lake Chad basin until 1991. Since then, cholera outbreaks have been reported every couple of years. The objective of this study was to investigate the 2010/2011 Vibrio cholerae outbreak in Cameroon to gain insight into the genomic...... make-up of the V. cholerae strains responsible for the outbreak. Twenty-four strains were isolated and whole genome sequenced. Known virulence genes, resistance genes and integrating conjugative element (ICE) elements were identified and annotated. A global phylogeny (378 genomes) was inferred using...... a single nucleotide polymorphism (SNP) analysis. The Cameroon outbreak was found to be clonal and clustered distant from the other African strains. In addition, a subset of the strains contained a deletion that was found in the ICE element causing less resistance. These results suggest that V. cholerae...

  3. Color correlation for the recognition of Vibrio cholerae O1 in seawater

    Science.gov (United States)

    Mourino-Perez, Rosa R.; Alvarez-Borrego, Josue

    1999-07-01

    Application of color correlation optical systems for the recognition of Vibrio cholerae 01 in seawater samples with matched filters and phase only filters recorded in holographic plates in three channels (RGB).

  4. DETECTION OF Vibrio choleraeO1 SEROTYPE IN ICE PRESERVATIVES OF SEA PRODUCTS AT KEDONGANAN FISH MARKET, KUTA, BALI

    Directory of Open Access Journals (Sweden)

    IGM Wijaya P

    2013-09-01

    Full Text Available Vibrio cholerae is one of gut attacking bacteria in which 20% of the victims suffered acute diarrhea and 10-20% of them were severe, in addition the mode of case in Indonesia are increasing. Bali is a threatened area especially by ice preservative of sea products investement since it has a tropical climate with tons of sea products, and one of most popular fish market is Kedonganan Fish Market. We were conducting a descriptive observational with explorative study which is aimed to detect V. cholerae investation in ice preservative. The samples are ice or melted ice which are used to preserve sea product at Kedonganan Fish Market and each were choosed randomly. The samples processed in Alkaline Peptone Water (APW and planted on TCBS media for microbiological culture exploration which refer to Kobe University protocol of V. cholerae isolation from environment; painted for gram evaluation based on our departement’s procedures; and latex serology evaluation using tools of V. cholerae O1 AD ‘seiken’. Of 10 samples obtained, 21 single bacterial colonies were found, in which 8 of them were suspected as V. cholerae’s colonies based on microbiological culture and gram painting evaluation. The suspected colonies are then undergone latex serology test to make sure the existence of V. cholerae and knowing its serotype. Based on the result analysis and interpretation of microbiological culture test, gram painting, and serology exploration, its found that 50% of samples were invested by Inaba type of V. cholerae O1. Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; text-align:justify; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt

  5. The neuraminidase gene is present in the non-toxigenic Vibrio cholerae Amazonia strain: a different allele in comparison to the pandemic strains

    Directory of Open Access Journals (Sweden)

    Sonia Catarina de Abreu Figueiredo

    2005-10-01

    Full Text Available The neuraminidase gene, nanH, is present in the O1, non-toxigenic Vibrio cholerae Amazonia strain. Its location has been assigned to a 150 kb NotI DNA fragment, with the use of pulsed-field gel electrophoresis and DNA hybridization. This NotI fragment is positioned inside 630 kb SfiI and 1900 kb I-CeuI fragments of chromosome 1. Association of the pathogenicity island VPI-2, carrying nanH and other genes, with toxigenic strains has been described by other authors. The presence of nanH in a non-toxigenic strain is an exception to this rule. The Amazonia strain nanH was sequenced (Genbank accession No. AY825932 and compared to available V. cholerae sequences. The sequence is different from those of pandemic strains, with 72 nucleotide substitutions. This is the first description of an O1 strain with a different nanH allele. The most variable domain of the Amazonia NanH is the second lectin wing, comprising 13 out of 17 amino acid substitutions. Based on the presence of nanH in the same region of the genome, and similarity of the adjacent sequences to VPI-2 sequences, it is proposed that the pathogenicity island VPI-2 is present in this strain.

  6. Genotypes associated with virulence in environmental isolates of O1/O139 Vibrio cholerae in Guangdong province%广东省外环境O1/O139群霍乱弧菌毒力基因分型

    Institute of Scientific and Technical Information of China (English)

    李柏生; 谭海玲; 邓小玲; 柯碧霞; 陈经雕; 刘美真; 何冬梅; 柯昌文

    2012-01-01

    目的 分析广东省外环境来源O1/O139群霍乱弧菌毒力基因的携带及基因分型特征,为霍乱防控提供依据.方法 选取2008 -2009年广东省O1/O139群霍乱弧菌水体分离株69株,水产品分离株16株和同期病例分离株5株,应用多重聚合酶链反应对ctxA、ace、zot、tcpA、tcpl、hlyA、ompU、toxR等8种毒力基因进行检测和分型分析.结果 90株O1/O139群霍乱弧菌均携带hlyA和toxR基因;5株病例菌株中有3株携带8种毒力基因,另外2株小川型菌株为非产毒株,基因型为hlyA+ toxR+ ompU+ zot+ tcpA+ tcpl+型和hlyA+ toxR+ tcpA+型;水体菌株中,稻叶型菌株以hlyA+ toxR+ ompU+ ace+ zot+ tcpl+型(34.15%)为主,小川型(66.67%)和O139群(70%)以hlyA+ toxR+型为主;水产品菌株中,稻叶型菌株以hlyA+ toxR+ ompU+ tcpl+型(75.00%)为主,小川型菌株各种基因型别均有分布,无明显优势基因型别.结论 广东省外环境来源O1/O139群霍乱弧菌以非产毒株广泛存在,毒力基因型别多样.%Objective To analyze the virulence genes and genotyping characteristics of environmental O1/O139 Vibrio cholerae{ V. cholerae) in Guangdong province and to provide the basis for the prevention and control of cholera. Methods Eight pairs of primers were designed according to ctxA,ace,zot,tcpA ,tcpl,hlyA ,ompU,and toxR. The multiplex PCR(MPCR) was established to detect 90 V. cholerae O1/O139 strains isolated between 2008 and 2009(69 aquatic strains, 16 sea food strains and 5 clinical strains). Genotypes associated with the virulence were determined then according to the result of the MPCR. Results The hlyA and toxR genes were positive in all the isolates. Three of five clinical isolates were detected for eight virulence genes and the other two isolates displayed the genotype of virulence with taxR+ ,ompU+ ,zot+ ,tcpA+ ,tcpl+ ,hlyA+ toxR+ ,and tcpA+. For the aquatic isolates, 14 Inaba strains(34. 15% ,14/41) were hlyA+ ,toxR+ ,ompU+ ,ace+ ,zot+ ,tcpI+ .while

  7. A Cholera Conjugate Vaccine Containing O-specific Polysaccharide (OSP of V. cholerae O1 Inaba and Recombinant Fragment of Tetanus Toxin Heavy Chain (OSP:rTTHc Induces Serum, Memory and Lamina Proprial Responses against OSP and Is Protective in Mice.

    Directory of Open Access Journals (Sweden)

    Md Abu Sayeed

    Full Text Available Vibrio cholerae is the cause of cholera, a severe watery diarrhea. Protection against cholera is serogroup specific. Serogroup specificity is defined by the O-specific polysaccharide (OSP component of lipopolysaccharide (LPS.Here we describe a conjugate vaccine for cholera prepared via squaric acid chemistry from the OSP of V. cholerae O1 Inaba strain PIC018 and a recombinant heavy chain fragment of tetanus toxin (OSP:rTTHc. We assessed a range of vaccine doses based on the OSP content of the vaccine (10-50 μg, vaccine compositions varying by molar loading ratio of OSP to rTTHc (3:1, 5:1, 10:1, effect of an adjuvant, and route of immunization.Immunized mice developed prominent anti-OSP and anti-TT serum IgG responses, as well as vibriocidal antibody and memory B cell responses following intramuscular or intradermal vaccination. Mice did not develop anti-squarate responses. Intestinal lamina proprial IgA responses targeting OSP occurred following intradermal vaccination. In general, we found comparable immune responses in mice immunized with these variations, although memory B cell and vibriocidal responses were blunted in mice receiving the highest dose of vaccine (50 μg. We found no appreciable change in immune responses when the conjugate vaccine was administered in the presence or absence of immunoadjuvant alum. Administration of OSP:rTTHc resulted in 55% protective efficacy in a mouse survival cholera challenge model.We report development of an Inaba OSP:rTTHc conjugate vaccine that induces memory responses and protection against cholera in mice. Development of an effective cholera conjugate vaccine that induces high level and long-term immune responses against OSP would be beneficial, especially in young children who respond poorly to polysaccharide antigens.

  8. Analysis of Immune Responses and Serological Cross Reactivities among Vibrio cholerae O1,Shigella flexneri 2a and Haemophilus influenzae b

    Institute of Scientific and Technical Information of China (English)

    Fazle Rabbi; Chowdhury R.Ahsan; Nasreen Sultana; Tasmina Rahman; H.M.Al-Emran; M.Nizam Uddin; Mahbub Hossain; Kazi Selim Anwar; Mahmuda Yasmin; Jamalun Nessa

    2008-01-01

    Antigenic determinants expressed on the bacterial cell surface are of importance in the serological characterization and microbiological diagnosis.The bacterial strains carrying these identical or similar antigenic epitopes might react with antibodies produced against other strains.In this study,strong immunogenicity and antigenic cross reactivity were demonstrated among V.choleae O1,S.flexnerii 2a and H.influenzae b surface components.The enzyme linked immunosorbent assay (ELISA) results were supported by Western blot analysis,where at least 20 antigenic bands,were obtained in each of the reactions,when the surface components were reacted with the homologous antisera.The indirect ELISA results also demonstrated high degree of antigenic relatedness between the surface components of these species,where each surface component was reacted with the heterologous antisera. Western blot analysis also revealed cross reactions between the surface components suggesting common distribution of antigensepitopes in these bacteriaI species.This study,thus,gave a clear idea of the level of antigenic sharing and variations among the pathogenic V.cholerae O1,S.flexneri 2a and H.influenzae b strains, which in future,may help in selecting a proper candidate for vaccines and immunodiagnostics development.

  9. Immune adjuvant effect of V. cholerae O1 derived Proteoliposome coadministered by intranasal route with Vi polysaccharide from Salmonella Typhi.

    Science.gov (United States)

    Acevedo, Reinaldo; Callicó, Adriana; Aranguren, Yisabel; Zayas, Caridad; Valdés, Yolanda; Pérez, Oliver; García, Luis; Ferro, Valerie A; Pérez, José Luis

    2013-01-01

    The proteoliposome derived from Vibrio cholerae O1 (PLc) is a nanoscaled structure obtained by a detergent extraction process. Intranasal (i.n) administration of PLc was immunogenic at mucosal and systemic level vs. V. cholerae; however the adjuvant potential of this structure for non-cholera antigens has not been proven yet. The aim of this work was to evaluate the effect of coadministering PLc with the Vi polysaccharide antigen (Poli Vi) of S. Typhi by the i.n route. The results showed that Poli Vi coadministered with PLc (PLc+Poli Vi) induce a higher IgA response in saliva (p0.05) to that induced in a group of mice immunised by the parenteral route with the Cuban anti-typhoid vaccine vax-TyVi, although this vaccine did not induce a mucosal response. In conclusion, this work demonstrates that PLc can be used as a mucosal adjuvant to potentiate the immune response against a polysaccharide antigen like Poli Vi.

  10. Cholera in Azov area

    OpenAIRE

    O. N. Domashenko; T. A. Belomerya; N. V. Martynova; G. N. Daragan; Demkovich, O.O.; U. V. Malakhova; G. I. Zemlyanskaya; Popova, D.M.

    2015-01-01

    The purpose of research is analysis of clinical course and treatment results of patients with cholera in the Azov area. Materials and methods. During the period from 29.05.2011 to 19.08.2011 33 cases of cholera (32 adults and 1 child) and 25 vibrio carriers (22 adults and 3 children), which were caused by toxigenic strains of Vibrio cholera El Tor serogroup O1 Ogawa. Results. Likely factors of disease transmission in Mariupol are sea and river water, and the fish that were caught in the water...

  11. Swedish isolates of Vibrio cholerae enhance their survival when interacted intracellularly with Acanthamoeba castellanii

    Directory of Open Access Journals (Sweden)

    Salah Shanan

    2016-04-01

    Full Text Available Vibrio cholerae is a Gram-negative bacterium that occurs naturally in aquatic environment. Only V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera, other serogroups can cause gastroenteritis, open wounds infection, and septicaemia. V. cholerae O1 and V. cholerae O139 grow and survive inside Acanthamoeba castellanii. The aim of this study is to investigate the interactions of the Swedish clinical isolates V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11, and V. cholerae O160 with A. castellanii. The interaction between A. castellanii and V. cholerae strains was studied by means of amoeba cell counts, viable counts of the bacteria in the absence or presence of amoebae, and of the intracellularly growing bacteria, visualised by electron microscopy. These results show that all V. cholerae can grow and survive outside and inside the amoebae, disclosing that V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11, and V. cholerae O160 all can be considered as facultative intracellular bacteria.

  12. Swedish isolates of Vibrio cholerae enhance their survival when interacted intracellularly with Acanthamoeba castellanii.

    Science.gov (United States)

    Shanan, Salah; Bayoumi, Magdi; Saeed, Amir; Sandström, Gunnar; Abd, Hadi

    2016-01-01

    Vibrio cholerae is a Gram-negative bacterium that occurs naturally in aquatic environment. Only V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera, other serogroups can cause gastroenteritis, open wounds infection, and septicaemia. V. cholerae O1 and V. cholerae O139 grow and survive inside Acanthamoeba castellanii. The aim of this study is to investigate the interactions of the Swedish clinical isolates V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11, and V. cholerae O160 with A. castellanii. The interaction between A. castellanii and V. cholerae strains was studied by means of amoeba cell counts, viable counts of the bacteria in the absence or presence of amoebae, and of the intracellularly growing bacteria, visualised by electron microscopy. These results show that all V. cholerae can grow and survive outside and inside the amoebae, disclosing that V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11, and V. cholerae O160 all can be considered as facultative intracellular bacteria.

  13. Phenotypic, Genotypic, and Antibiotic Sensitivity Patterns of Strains Isolated from the Cholera Epidemic in Zimbabwe

    NARCIS (Netherlands)

    Islam, Mohammad S.; Mahmud, Zahid H.; Ansaruzzaman, Mohammad; Faruque, Shah M.; Talukder, Kaisar A.; Qadri, Firdausi; Alam, Munirul; Islam, Shafiqul; Bardhan, Pradip K.; Mazumder, Ramendra N.; Khan, Azharul I.; Ahmed, Sirajuddin; Iqbal, Anwarul; Chitsatso, Owen; Mudzori, James; Patel, Sheetal; Midzi, Stanley M.; Charimari, Lincoln; Endtz, Hubert P.; Cravioto, Alejandro

    2011-01-01

    This paper details the phenotypic, genotypic, and antibiotic sensitivity patterns of 88 Vibrio cholerae strains from Zimbabwe. Of the 88 strains, 83 were classified as "altered El Tor" and 5 as "hybrid El Tor" strains. All of the strains were susceptible to tetracycline, doxycycline, ciprofloxacin,

  14. gbpA as a Novel qPCR Target for the Species-Specific Detection of Vibrio cholerae O1, O139, Non-O1/Non-O139 in Environmental, Stool, and Historical Continuous Plankton Recorder Samples.

    Science.gov (United States)

    Vezzulli, Luigi; Stauder, Monica; Grande, Chiara; Pezzati, Elisabetta; Verheye, Hans M; Owens, Nicholas J P; Pruzzo, Carla

    2015-01-01

    The Vibrio cholerae N-acetyl glucosamine-binding protein A (GbpA) is a chitin-binding protein involved in V. cholerae attachment to environmental chitin surfaces and human intestinal cells. We previously investigated the distribution and genetic variations of gbpA in a large collection of V. cholerae strains and found that the gene is consistently present and highly conserved in this species. Primers and probe were designed from the gbpA sequence of V. cholerae and a new Taq-based qPCR protocol was developed for diagnostic detection and quantification of the bacterium in environmental and stool samples. In addition, the positions of primers targeting the gbpA gene region were selected to obtain a short amplified fragment of 206 bp and the protocol was optimized for the analysis of formalin-fixed samples, such as historical Continuous Plankton Recorder (CPR) samples. Overall, the method is sensitive (50 gene copies), highly specific for V. cholerae and failed to amplify strains of the closely-related species Vibrio mimicus. The sensitivity of the assay applied to environmental and stool samples spiked with V. cholerae ATCC 39315 was comparable to that of pure cultures and was of 102 genomic units/l for drinking and seawater samples, 101 genomic units/g for sediment and 102 genomic units/g for bivalve and stool samples. The method also performs well when tested on artificially formalin-fixed and degraded genomic samples and was able to amplify V. cholerae DNA in historical CPR samples, the earliest of which date back to August 1966. The detection of V. cholerae in CPR samples collected in cholera endemic areas such as the Benguela Current Large Marine Ecosystem (BCLME) is of particular significance and represents a proof of concept for the possible use of the CPR technology and the developed qPCR assay in cholera studies.

  15. gbpA as a Novel qPCR Target for the Species-Specific Detection of Vibrio cholerae O1, O139, Non-O1/Non-O139 in Environmental, Stool, and Historical Continuous Plankton Recorder Samples.

    Directory of Open Access Journals (Sweden)

    Luigi Vezzulli

    Full Text Available The Vibrio cholerae N-acetyl glucosamine-binding protein A (GbpA is a chitin-binding protein involved in V. cholerae attachment to environmental chitin surfaces and human intestinal cells. We previously investigated the distribution and genetic variations of gbpA in a large collection of V. cholerae strains and found that the gene is consistently present and highly conserved in this species. Primers and probe were designed from the gbpA sequence of V. cholerae and a new Taq-based qPCR protocol was developed for diagnostic detection and quantification of the bacterium in environmental and stool samples. In addition, the positions of primers targeting the gbpA gene region were selected to obtain a short amplified fragment of 206 bp and the protocol was optimized for the analysis of formalin-fixed samples, such as historical Continuous Plankton Recorder (CPR samples. Overall, the method is sensitive (50 gene copies, highly specific for V. cholerae and failed to amplify strains of the closely-related species Vibrio mimicus. The sensitivity of the assay applied to environmental and stool samples spiked with V. cholerae ATCC 39315 was comparable to that of pure cultures and was of 102 genomic units/l for drinking and seawater samples, 101 genomic units/g for sediment and 102 genomic units/g for bivalve and stool samples. The method also performs well when tested on artificially formalin-fixed and degraded genomic samples and was able to amplify V. cholerae DNA in historical CPR samples, the earliest of which date back to August 1966. The detection of V. cholerae in CPR samples collected in cholera endemic areas such as the Benguela Current Large Marine Ecosystem (BCLME is of particular significance and represents a proof of concept for the possible use of the CPR technology and the developed qPCR assay in cholera studies.

  16. Epidemic tvpe and antibiotic resistance of cholera strains isolated%霍乱疫情分离株流行菌型及耐药性分析

    Institute of Scientific and Technical Information of China (English)

    王少玲; 吴捷; 刁保卫; 崔志刚; 王衍德; 朱建华; 黄昌和; 马焱

    2012-01-01

    Objective To assess the epidemic type and antibiotic resistance of cholere strains isolated and to provide basis for clinical treatment and prevention of cholera. Methods Six samples from an outbreak of cholera epidemic were collected in Hainan province in June 2008 and analyzed by serotyping and phage-biotyping. The cholera toxin gene was detected with PCR according to the " Manual for the Prevention and Control of Cholera" (version 5). Pulsed field gel electro-phoresis( PFGE) was used for subtyping of the isolates. The antibiotic susceptibility test of the 6 strains to 17 kinds of antibacterial was determined with improved K-B method recommended by WHO. Results The 6 strains were confirmed as Vibrio cholerae El Tor biotype, with 5 strains of 01 group of Vibrio cholerae El Tor serotype Inaba lc and 1 strains was Vibrio cholerae Ogawa( lc). The cholera toxin gene was identified in all 6 strains. The mapping of the strains were belonged to two pattern by PFGE and the similarity was 100 %. The 6 strains were all resistant to streptomycin,sulphamethoxazole/trim-ethoprim, sulphafurazole, and polymyxin B. High drug susceptibility of the samples was found to 9 kinds of antibiotics such as norfloxacin.cefotaxime and cephalothin(53% ,9/17). Conclusion The bacterium type of Vibrio cholerae causing the epidemic outbreak of cholera was Ol group of Vibrio cholerae El Tor serotype Inaba lc. Norfloxacin.cefotaxime and cepha-lothin could be the choice of clinical treatment for the cases and carriers of Vibrio cholerae in Hainan province. But streptomycin, sulphamethoxazole/trimethoprim,sulphafurazole,and polymyxin B could not be used.%目的 了解霍乱疫情分离株的流行菌型及其耐药性,为霍乱疫情的快速有效控制和临床治疗提供科学依据.方法 对2008年6月海南省某地霍乱疫情采集的6株霍乱弧菌分离株进行血清学分型、噬菌体-生物分型、霍乱毒素基因检测、脉冲场凝胶电泳(PFGE)分子分型

  17. Vibrio cholerae O1 em amostras de ambientes aquáticos e de alimentos analisados no Estado de Pernambuco, Brasil

    OpenAIRE

    Colaço Waldêny; Silva Filho Sandoval Vieira da; Rodrigues Dália dos Prazeres; Hofer Ernesto

    1998-01-01

    No período de 1992 a 1994, foram analisadas 2.585 amostras de águas de diferentes ecossistemas, acrescidas de 91 espécimens de alimentos visando ao monitoramento de Vibrio cholerae O1 no Estado de Pernambuco. Nas 2.676 amostras foram detectadas 193 cepas de Vibrio cholerae O1 (7,21%) com predominância do sorovar Inaba (183-94,8%) sobre Ogawa (10-5,1%), todas classificadas no biotipo El Tor e sensíveis à tetraciclina. Numa parcela de setenta amostras selecionadas ao acaso, mas incluindo todas ...

  18. Establishment of multiplex real-time quantitative PCR method for rapid detection of toxigenic Vibrio cholerae serogroup O1 from environmental sample%外环境样本中产毒型O1群霍乱弧菌双重荧光定量PCR快速检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    黄世旺; 徐丹戈; 徐昌平; 张政; 方叶珍; 包芳珍; 李剑; 蒋雪凤; 卢亦愚

    2011-01-01

    To establish a TaqMan-based multiplex real-time PCR assay for detection of toxigenic Vibrio cholerae serogroup O1 and construct a method for primary application of environmental samples test, the gene sequences of cholera toxin and specific O antigen biosynthetic gene rrb of serogroup O1 downloaded from the GenBank were aligned using the biologic software, and the specific primers and probe were designed in the conserved region of the CT and rfb-O1 gene for Vibrio cholerae serogroup O1. The reaction conditions were optimized and the sensitivity, specificity and the stability of the assay were evaluated. The clinical specimens collected from the environment were detected by this assay. For specifically detecting the toxigenic Vibrio cholerae serogroup O1, the detection limits of the assay for rfb-O1 and CT gene were 100 cfu/mL and the regression coefficient of the quantitative curve were 0. 998 and 0. 999, respectively. Five strains of toxigenic Vibrio cholerae serogroup O1 were collected from 352 environmental specimens for the first time by this assay. This assay is a rapid, sensitive and specific one for the widespread detection of toxigenic Vibrio cholerae serogroup O1 from environmental sample.%目的 研究建立双重荧光定量PCR技术快速检测产毒型O1群霍乱弧菌,并首次应用于外环境样本的检测中.方法 从GenBank上下载O1群霍乱弧菌O抗原编码基因rfbO1和毒力基因CT序列,在rfbO1和CT保守区域设计特异性引物和探针,建立优化单一和双重荧光PCR反应体系,评价所建双重PCR反应体系的特异性、敏感性和稳定性,并应用于外环境样本的监测检验中.结果 该方法对O1群霍乱弧菌检测具有高度特异性,对rfbO1和CT基因序列检出限达到1.0×102cfu/mL,构建的体系定量标准曲线相关系数分别为0.999和0.998,具有较好的稳定性,并首次从352件外环境样本中检测出了5株产毒型O1群霍乱弧菌.结论 本研究建立的双重荧光定量PCR

  19. 广州海珠地区非O1/非O139群霍乱弧菌流行状况调查及生物学特征研究%Epidemic condition and biological characteristics of non-O1/non-O139 vibrio cholerae in Haizhu District of Guangzhou

    Institute of Scientific and Technical Information of China (English)

    许少洪; 李映霞; 李少彤; 吴琪; 孙凤琪; 黄芳; 曾爱芳

    2010-01-01

    Objective To understand the epidemic condition, distribution and biological characteristics of non-O1/non-O139 vibrio cholerae from 2001 to 2009 in Haizhu District, to provide a scientific basis for the prevention and control of acute diarrhea. Methods Referring to the detecting method written in "Cholera control handbook"in the fifth edition,764 specimens from outside environment (including the water in the Pearl River, drinking water, water for breeding fish, aquatic products and delicatessen foods), 189 specimens of healthy population and 3398 intestinal samples of patients with diarrhea, summing up to 4351 specimens for non-O1/non-O139 vibrio cholerae test. Results 4351 speciments were detected of 101 strains of non O1/non O139 vibrio cholerae, the total detection rate was 2. 32%;66 strains were identified by serotyping and grouped into 26 different serotypes, the typing rate was 65.3%. The strains VBO9,VBO38 and VBO76 were the dominant bacteria. Nine strains of the same type of non-O1/non-O139 vibrio cholerae were found from external environments also from patients with diarrhea, suggesting that there might be a correlation between the two. Conclusion Non-O1/non-O139 vibrio cholerae have diversified serotypes, causing certain infection rate among the population in this region. These bacteria exist extensively in external environment and they are the potential hazard to the citizens.%目的 了解广州海珠地区2001-2009年非O1/非O139群霍乱弧菌的流行、分布状况及生物学特征,为防控该类病原菌引起的急性腹泻病提供科学依据.方法 参照(第5版)中的检测方法,对本区珠江河水、饮用水、养殖水、各种海(水)产品、市售熟食等外环境标本764份、健康人群标本189份及肠道门诊腹泻患者标本3398份,共计4351份标本进行非O1/非O139群霍乱弧菌的检定.结果 4351份标本共检出非O1/非O139群霍乱弧菌101株,总检出率为2.32%;其中有66株可分型,分属于26

  20. Detection of ctx gene positive non-O1/non-O139 V. cholerae in shrimp aquaculture environments

    OpenAIRE

    Madhusudana, Rao B.; Surendran, P. K.

    2011-01-01

    Water and post-larvae samples from black tiger (Penaeus monodon) shrimp hatcheries; pond water, pond sediment and shrimp from aquaculture farms were screened for the presence of V. cholerae. A V. cholerae-duplex PCR method was developed by utilizing V. cholerae species specific sodB primers and ctxAB genes specific primers. Incidence of V. cholerae was not observed in shrimp hatchery samples but was noticed in aquaculture samples. The incidence of V. cholerae was higher in pond water (7.6%) t...

  1. [Multilocus sequence-typing of vibrio cholerae strains with various epidemic importance].

    Science.gov (United States)

    Mironova, L V; Afanas'ev, M V; Goldapel, E G; Balakhonov, S V

    2015-01-01

    The allele polymorphism of the housekeeping genes (dnaE, lap, recA, pgm, gyrB, cat, chi, gmd) from the Vibrio cholerae strains with different epidemic importance (n = 41) isolated in Siberia and at the Far East during the cholera pandemic VII was tested. All toxigenic strains isolated at the period of epidemic complications irrespective of time and source of isolation were characterized by the identical allele profile and belonged to the same sequence-type. Nine sequence types were detected in non-epidemic isolates. The dendrogram clustering was associated with the serogroup and in some cases with the territory and time of isolation. The structure heterogeneity of the non-toxigenic V. cholerae housekeeping genes was in most cases caused by the synonymous nucleotide replacements (Dn/Ds cholerae at the analyzed genome sites. The revealed distinctions in the structure of housekeeping genes of the V. cholerae with different epidemic importance can be regarded as evidence of various evolutional directions in these strain groups.

  2. IncA/C Conjugative Plasmids Mobilize a New Family of Multidrug Resistance Islands in Clinical Vibrio cholerae Non-O1/Non-O139 Isolates from Haiti

    Directory of Open Access Journals (Sweden)

    Nicolas Carraro

    2016-07-01

    Full Text Available Mobile genetic elements play a pivotal role in the adaptation of bacterial populations, allowing them to rapidly cope with hostile conditions, including the presence of antimicrobial compounds. IncA/C conjugative plasmids (ACPs are efficient vehicles for dissemination of multidrug resistance genes in a broad range of pathogenic species of Enterobacteriaceae. ACPs have sporadically been reported in Vibrio cholerae, the infectious agent of the diarrheal disease cholera. The regulatory network that controls ACP mobility ultimately depends on the transcriptional activation of multiple ACP-borne operons by the master activator AcaCD. Beyond ACP conjugation, AcaCD has also recently been shown to activate the expression of genes located in the Salmonella genomic island 1 (SGI1. Here, we describe MGIVchHai6, a novel and unrelated mobilizable genomic island (MGI integrated into the 3′ end of trmE in chromosome I of V. cholerae HC-36A1, a non-O1/non-O139 multidrug-resistant clinical isolate recovered from Haiti in 2010. MGIVchHai6 contains a mercury resistance transposon and an integron In104-like multidrug resistance element similar to the one of SGI1. We show that MGIVchHai6 excises from the chromosome in an AcaCD-dependent manner and is mobilized by ACPs. Acquisition of MGIVchHai6 confers resistance to β-lactams, sulfamethoxazole, tetracycline, chloramphenicol, trimethoprim, and streptomycin/spectinomycin. In silico analyses revealed that MGIVchHai6-like elements are carried by several environmental and clinical V. cholerae strains recovered from the Indian subcontinent, as well as from North and South America, including all non-O1/non-O139 clinical isolates from Haiti.

  3. Molecular Analysis and Toxigenic Potential of Vibrio cholerae Isolated from Hilsha fish (Tenualosa ilisha), Bangladesh

    DEFF Research Database (Denmark)

    Hossain, Zenat Zebin; Farhana, Israt; Tulsiani, Suhella

    Exposure to contaminated fish may upsurge the virulent strains of Vibrio cholerae, the deadly human pathogen in the households of rural and urban Bangladesh. Since V. cholerae spreading was reported from the Bay of Bengal, this study hypothesized that Hilsha (Tenualosa ilisha), a marine and fresh...... water fish may serve as a transmission vehicle of potential emerging epidemic causing strains. For this, we studied 9 toxigenic V. cholerae strains isolated from Hilsha fish including 6 V. cholerae O1 and 3 non O1/O139 serogroups for virulence associated genotype and their pathogenic potential on animal...... model and human cancer cell line . The V. cholerae O1 and non- O1/O139 strains possessed diverse virulence genes but lacked some major toxin genes like ctxA, tcp etc. Eight of the nine strains showed survivability up to 1% sodium chloride in broth culture which indicates their coastal origin. All nine...

  4. Development of a Multiplex PCR for Discrimination of the TLC:RS1:CTX array of Vibrio cholerae Wave 3 El Tor Strains.

    Science.gov (United States)

    Kim, Eun Jin; Yu, Hyun Jin; Nair, G Balakrish; Kim, Dong Wook

    2016-12-28

    Vibrio cholerae O1 serogroup Wave 3 El Tor strains are presently prevalent worldwide. The Wave 3 El Tor strains contain a TLC:RS1:CTX array on chromosome 1, and no element is integrated on chromosome 2. A multiplex PCR optimized to identify the TLC:RS1:CTX array of Wave 3 strains has been developed in this study. By using eight primers, the multiplex PCR can identify the characteristic CTX and RS1 array of Wave 3 strains from various arrays of strains belonging to other Waves. The four amplified DNA fragments of Wave 3 strains have been cloned in a vector, which could be used as a positive control for the multiplex PCR. This multiplex PCR and the positive control set could be useful tools for rapid recognition of Wave 3 El Tor strains.

  5. Detection of vibrio cholerae O1 by using cerium oxide nanowires - based immunosensor with different antibody immobilization methods

    Science.gov (United States)

    Tam, Phuong Dinh; Hoang, Nguyen Luong; Lan, Hoang; Vuong, Pham Hung; Anh, Ta Thi Nhat; Huy, Tran Quang; Thuy, Nguyen Thanh

    2016-05-01

    In this work, we evaluated the effects of different antibody immobilization strategies on the response of a CeO2-nanowires (NWs)-based immunosensor for Vibrio cholerae O1 detection. Accordingly, the changes in the electron-transfer resistance ( R et ) from before to after cells bind to an antibody-modified electrode prepared by using three different methods of antibody immobilization were determined. The values were 16.2%, 8.3%, and 6.65% for the method that utilized protein A, antibodies activated by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS), and absorption, respectively. Cyclic voltammetry confirmed that the change in the current was highest for the immunosensors prepared using protein A (11%), followed by those prepared with EDC/NHS-activated antibodies (9%), and finally, those prepared through absorption (7.5%). The order of the antibody immobilization strategies in terms of resulting immunosensor detection limit and sensitivity was as follows order: absorption (3.2 × 103 CFU/mL; 45.1 Ω/CFU·mL-1) < EDC/NHS-activated antibody (1.0 × 103 CFU/mL; 50.6 Ω/CFU·mL-1) < protein A (1.0 × 102 CFU/mL; 65.8 Ω/CFU·mL-1). Thus, we confirmed that the protein A - mediated method showed significantly high cell binding efficiencies compared to the random immobilization method.

  6. A physical map of the chromosomal region determining O-antigen biosynthesis in Vibrio cholerae O1.

    Science.gov (United States)

    Ward, H M; Morelli, G; Kamke, M; Morona, R; Yeadon, J; Hackett, J A; Manning, P A

    1987-01-01

    We have previously described the cosmid cloning of the genes determining the biosynthesis of the Inaba and Ogawa O-antigens of the lipopolysaccharides of Vibrio cholerae O1 (Manning et al., 1986). By Southern hybridization analysis of chromosomal and cosmid DNA, and heteroduplex analysis between the clones we have been able to precisely define the region of contiguous chromosomal DNA in the vicinity of the O-antigen-encoding region. These data and comparison of end points of clones and of deletion derivatives demonstrate that at least 16 kb of a 19-kb SstI fragment is required to encode O-antigen biosynthesis. Expression of O-antigen is independent of the orientation of this SstI fragment with respect to cloning vectors suggesting that its regulatory region has been cloned intact. No detectable differences were observed in the restriction patterns of the Inaba and Ogawa coding regions implying that only minor changes are involved when serotype conversion (Inaba to Ogawa or vice versa) occurs. Bhaskaran [Ind. J. Med. Res. 47 (1959) 253-260] originally defined this region associated with O-antigen biosynthesis oag; however, to be consistent with other organisms [Hitchcock et al., J. Bacteriol. 166 (1986) 699-705], it is suggested this be changed to rfb.

  7. Phylogenetic Diversity of Vibrio cholerae Associated with Endemic Cholera in Mexico from 1991 to 2008

    Directory of Open Access Journals (Sweden)

    Seon Young Choi

    2016-03-01

    Full Text Available An outbreak of cholera occurred in 1991 in Mexico, where it had not been reported for more than a century and is now endemic. Vibrio cholerae O1 prototype El Tor and classical strains coexist with altered El Tor strains (1991 to 1997. Nontoxigenic (CTX− V. cholerae El Tor dominated toxigenic (CTX+ strains (2001 to 2003, but V. cholerae CTX+ variant El Tor was isolated during 2004 to 2008, outcompeting CTX−V. cholerae. Genomes of six Mexican V. cholerae O1 strains isolated during 1991 to 2008 were sequenced and compared with both contemporary and archived strains of V. cholerae. Three were CTX+ El Tor, two were CTX− El Tor, and the remaining strain was a CTX+ classical isolate. Whole-genome sequence analysis showed the six isolates belonged to five distinct phylogenetic clades. One CTX− isolate is ancestral to the 6th and 7th pandemic CTX+V. cholerae isolates. The other CTX− isolate joined with CTX− non-O1/O139 isolates from Haiti and seroconverted O1 isolates from Brazil and Amazonia. One CTX+ isolate was phylogenetically placed with the sixth pandemic classical clade and the V. cholerae O395 classical reference strain. Two CTX+ El Tor isolates possessing intact Vibrio seventh pandemic island II (VSP-II are related to hybrid El Tor isolates from Mozambique and Bangladesh. The third CTX+ El Tor isolate contained West African-South American (WASA recombination in VSP-II and showed relatedness to isolates from Peru and Brazil. Except for one isolate, all Mexican isolates lack SXT/R391 integrative conjugative elements (ICEs and sensitivity to selected antibiotics, with one isolate resistant to streptomycin. No isolates were related to contemporary isolates from Asia, Africa, or Haiti, indicating phylogenetic diversity.

  8. Cross feeding of glucose metabolism byproducts of Escherichia coli human gut isolates and probiotic strains affect survival of Vibrio cholerae.

    Science.gov (United States)

    Sengupta, Chirantana; Ekka, Manjula; Arora, Saurabh; Dhaware, Prashant D; Chowdhury, Rukhsana; Raychaudhuri, Saumya

    2017-01-01

    Vibrio cholerae converts glucose into either acid or the neutral end product acetoin and its survival in carbohydrate enriched media is linked to the nature of the byproducts produced. It has been demonstrated in this study that Escherichia coli strain isolated from the gut of healthy human volunteers and the commonly used probiotic E. coli Nissle strain that metabolize glucose to acidic byproducts drastically reduce the survival of V. cholerae strains irrespective of their glucose sensitivity and acetoin production status. Accordingly, E. coli glucose transport mutants that produce lower amounts of acidic metabolites had little effect on the survival of V. cholerae in cocultures. Thus, cross feeding of byproducts of glucose metabolism by heterologous bacteria modulates the survival of V. cholerae in glucose rich medium suggesting that composition of the gut microbiota could influence the outcome of V. cholerae infection especially when glucose based ORS is administered.

  9. O1/O139群霍乱弧菌检出率与珠江河口水体理化因素的相关性研究%The correlation study on vibrio cholerae O1/O139 detection rate and the physical and chemical factors of pearl river estuary water

    Institute of Scientific and Technical Information of China (English)

    许斌; 刘杰; 李柏生; 罗可天; 牟成惠

    2012-01-01

    Objective:To investigate the correlation between Vibro Cholerae 01/0139 detection rate and the physical and chemical factors of Pearl River Estuary water and provide a reference for cholera prevention and control. Methods: 24 sampling points along the Pearl River have been set to collect the Pearl River estuary water per month since 2010. With reference to Cholera Prevention and Control Manual (1999) by the Disease Control Division of the Ministry of Health, Vibrio cholerae was isolated for culture and identification. The ion density of water samples were tested by 7500a inductance coupled plasma mass spectrometry instrument. Results: 576 water samples were collected totally. 51 Vibrio choleras 01/0139 strains were detected and the detection rate was 8. 85% . The detection rate of Vibrio cholerae 01/0139 in summer accounted for more than 30% . Vibrio cholerae detection rate had linear relationship with Al3 + , Mn 2+ , Ni2 +. In the urban district and outside the Pearl River, the significant difference existed between the contents of Mg2+ ,K + ,Ca2+ ,Mn2+ ,Ni2+ ,Se4+ ,Sb3+ and PH value in water samples. Conclusion: The detection Vibrio cholerae of 01/0139 has some correlation to physical and chemical factors ( Al3+ , Mn2+. Ni2+ )of Pearl River estuary water.%目的:探讨珠江水中O1/O139群霍乱弧菌检出率与珠江水体中理化因素之间的相关性,为霍乱的防控工作提供参考.方法:2010年每月在珠江河沿岸24个采样点采集珠江河口水体,参照卫生部疾病控制司1999年版《霍乱防治手册》进行霍乱弧菌的分离培养和菌型鉴定,用7500a电感耦合等离子体质谱仪进行水样各离子浓度的检测.结果:共采集水样576份,检出51株O1/O139群霍乱弧菌,检出率为8.85%.夏天检出O1/O139群霍乱弧菌数占30%以上.Al、Mn及Ni与O1/O139群霍乱弧菌检出率之间有一定的线性关系.市内、外珠江水中Mg2+、K+、Ca2+、Mn2+、Ni2+、Se4+、Sb3+含量和PH值存在显著或

  10. Non-O1/Non-O139 Vibrio cholerae Avian Isolate from France Cocarrying the bla(VIM-1) and bla(VIM-4) Genes.

    Science.gov (United States)

    Aberkane, Salim; Compain, Fabrice; Barraud, Olivier; Ouédraogo, Abdoul-Salam; Bouzinbi, Nicolas; Vittecoq, Marion; Jean-Pierre, Hélène; Decré, Dominique; Godreuil, Sylvain

    2015-10-01

    We describe here a non-O1/non-O139 Vibrio cholerae isolate producing both VIM-1 and VIM-4 carbapenemases. It was isolated from a yellow-legged gull in southern France. The blaVIM genes were part of a class 1 integron structure located in an IncA/C plasmid. This study emphasizes the presence of carbapenemase genes in wildlife microbiota.

  11. High prevalence and diversity of pre-CTXΦ alleles in the environmental Vibrio cholerae O1 and O139 strains in the Zhujiang River estuary.

    Science.gov (United States)

    Wang, Duochun; Wang, Xiaomei; Li, Baisheng; Deng, Xiaoling; Tan, Hailing; Diao, Baowei; Chen, Jingdiao; Ke, Bixia; Zhong, Haojie; Zhou, Haijian; Ke, Changwen; Kan, Biao

    2014-06-01

    Toxigenic conversion of environmental Vibrio cholerae strains through lysogenic infection by the phage CTXΦ is an important step in the emergence of new pathogenic clones. The precursor form of the CTXΦ phage, pre-CTXΦ, does not carry the cholera toxin gene. During our investigation, we frequently found pre-CTXΦ prophages in non-toxigenic isolates in the serogroups of O1 and O139 strains in the Zhujiang estuary. We observed high amounts of sequence variation of rstR and gIII(CTX) in the pre-CTXΦ alleles as well as in the tcpA sequences within the strains. In addition, a new pre-CTXΦ allele, with a novel rstR sequence type and hybrid RS2, was identified. Our findings show that active, complicated gene recombination and horizontal transfer of pre-CTXΦs occurs within V. cholerae environmental strains, which creates a complex intermediate pool for the generation of toxigenic clones in the estuarine environment.

  12. Genome sequence of the human pathogen Vibrio cholerae Amazonia.

    NARCIS (Netherlands)

    Thompson, C.C.; Marin, M.A.; Dias, G.M.; Dutilh, B.E.; Edwards, R.A.; Iida, T.; Thompson, F.L.; Vicente, A.C.

    2011-01-01

    Vibrio cholerae O1 Amazonia is a pathogen that was isolated from cholera-like diarrhea cases in at least two countries, Brazil and Ghana. Based on multilocus sequence analysis, this lineage belongs to a distinct profile compared to strains from El Tor and classical biotypes. The genomic analysis rev

  13. Characterization and Genetic Variation of Vibrio cholerae Isolated from Clinical and Environmental Sources in Thailand.

    Science.gov (United States)

    Siriphap, Achiraya; Leekitcharoenphon, Pimlapas; Kaas, Rolf S; Theethakaew, Chonchanok; Aarestrup, Frank M; Sutheinkul, Orasa; Hendriksen, Rene S

    2017-01-01

    Cholera is still an important public health problem in several countries, including Thailand. In this study, a collection of clinical and environmental V. cholerae serogroup O1, O139, and non-O1/non-O139 strains originating from Thailand (1983 to 2013) was characterized to determine phenotypic and genotypic traits and to investigate the genetic relatedness. Using a combination of conventional methods and whole genome sequencing (WGS), 78 V. cholerae strains were identified. WGS was used to determine the serogroup, biotype, virulence, mobile genetic elements, and antimicrobial resistance genes using online bioinformatics tools. In addition, phenotypic antimicrobial resistance was determined by the minimal inhibitory concentration (MIC) test. The 78 V. cholerae strains belonged to the following serogroups O1: (n = 44), O139 (n = 16) and non-O1/non-O139 (n = 18). Interestingly, we found that the typical El Tor O1 strains were the major cause of clinical cholera during 1983-2000 with two Classical O1 strains detected in 2000. In 2004-2010, the El Tor variant strains revealed genotypes of the Classical biotype possessing either only ctxB or both ctxB and rstR while they harbored tcpA of the El Tor biotype. Thirty O1 and eleven O139 clinical strains carried CTXϕ (Cholera toxin) and tcpA as well four different pathogenic islands (PAIs). Beside non-O1/non-O139, the O1 environmental strains also presented chxA and Type Three Secretion System (TTSS). The in silico MultiLocus Sequence Typing (MLST) discriminated the O1 and O139 clinical strains from other serogroups and environmental strains. ST69 was dominant in the clinical strains belonging to the 7th pandemic clone. Non-O1/non-O139 and environmental strains showed various novel STs indicating genetic variation. Multidrug-resistant (MDR) strains were observed and conferred resistance to ampicillin, azithromycin, nalidixic acid, sulfamethoxazole, tetracycline, and trimethoprim and harboured variants of the SXT elements

  14. Characterization and Genetic Variation of Vibrio cholerae Isolated from Clinical and Environmental Sources in Thailand

    Science.gov (United States)

    Siriphap, Achiraya; Leekitcharoenphon, Pimlapas; Kaas, Rolf S.; Theethakaew, Chonchanok; Aarestrup, Frank M.; Sutheinkul, Orasa; Hendriksen, Rene S.

    2017-01-01

    Cholera is still an important public health problem in several countries, including Thailand. In this study, a collection of clinical and environmental V. cholerae serogroup O1, O139, and non-O1/non-O139 strains originating from Thailand (1983 to 2013) was characterized to determine phenotypic and genotypic traits and to investigate the genetic relatedness. Using a combination of conventional methods and whole genome sequencing (WGS), 78 V. cholerae strains were identified. WGS was used to determine the serogroup, biotype, virulence, mobile genetic elements, and antimicrobial resistance genes using online bioinformatics tools. In addition, phenotypic antimicrobial resistance was determined by the minimal inhibitory concentration (MIC) test. The 78 V. cholerae strains belonged to the following serogroups O1: (n = 44), O139 (n = 16) and non-O1/non-O139 (n = 18). Interestingly, we found that the typical El Tor O1 strains were the major cause of clinical cholera during 1983–2000 with two Classical O1 strains detected in 2000. In 2004–2010, the El Tor variant strains revealed genotypes of the Classical biotype possessing either only ctxB or both ctxB and rstR while they harbored tcpA of the El Tor biotype. Thirty O1 and eleven O139 clinical strains carried CTXϕ (Cholera toxin) and tcpA as well four different pathogenic islands (PAIs). Beside non-O1/non-O139, the O1 environmental strains also presented chxA and Type Three Secretion System (TTSS). The in silico MultiLocus Sequence Typing (MLST) discriminated the O1 and O139 clinical strains from other serogroups and environmental strains. ST69 was dominant in the clinical strains belonging to the 7th pandemic clone. Non-O1/non-O139 and environmental strains showed various novel STs indicating genetic variation. Multidrug-resistant (MDR) strains were observed and conferred resistance to ampicillin, azithromycin, nalidixic acid, sulfamethoxazole, tetracycline, and trimethoprim and harboured variants of the SXT elements

  15. Clinical isolates of Vibrio cholerae O1 El Tor Ogawa of 2009 from Kolkata, India: preponderance of SXT element and presence of Haitian ctxB variant.

    Directory of Open Access Journals (Sweden)

    Braj M R N S Kutar

    Full Text Available BACKGROUND: Increase in the number of multidrug resistant pathogens and the accompanied rise in case fatality rates has hampered the treatment of many infectious diseases including cholera. Unraveling the mechanisms responsible for multidrug resistance in the clinical isolates of Vibrio cholerae would help in understanding evolution of these pathogenic bacteria and their epidemic potential. This study was carried out to identify genetic factors responsible for multiple drug resistance in clinical isolates of Vibrio cholerae O1, serotype Ogawa, biotype El Tor isolated from the patients admitted to the Infectious Diseases Hospital, Kolkata, India, in 2009. METHODOLOGY/PRINCIPAL FINDINGS: One hundred and nineteen clinical isolates of V. cholerae were analysed for their antibiotic resistance phenotypes. Antibiogram analysis revealed that majority of the isolates showed resistance to co-trimoxazole, nalidixic acid, polymixin B and streptomycin. In PCR, SXT integrase was detected in 117 isolates and its sequence showed 99% identity notably to ICEVchInd5 from Sevagram, India, ICEVchBan5 from Bangladesh and VC1786ICE sequence from Haiti outbreak among others. Antibiotic resistance traits corresponding to SXT element were transferred from the parent Vibrio isolate to the recipient E. coli XL-1 Blue cells during conjugation. Double-mismatch-amplification mutation assay (DMAMA revealed the presence of Haitian type ctxB allele of genotype 7 in 55 isolates and the classical ctxB allele of genotype 1 in 59 isolates. Analysis of topoisomerase sequences revealed the presence of mutation Ser83 → Ile in gyrA and Ser85→ Leu in parC. This clearly showed the circulation of SXT-containing V. cholerae as causative agent for cholera in Kolkata. CONCLUSIONS: There was predominance of SXT element in these clinical isolates from Kolkata region which also accounted for their antibiotic resistance phenotype typical of this element. DMAMA PCR showed them to be a mixture

  16. Vibrio cholerae No O1 en muestras de aguas no cloradas consumidas por pobladores de las localidades de Santa y Coishco (Ancash, 2003 - 2004

    Directory of Open Access Journals (Sweden)

    Ana García P

    2006-07-01

    Full Text Available Objetivo: Identificar la presencia de Vibrio cholerae en muestras de agua no cloradas para consumo humano en las localidades de Santa y Coishco. Materiales y métodos: Entre julio de 2003 a junio de 2004 se tomaron muestras de agua, en forma semanal, provenientes de siete pozos con bomba manuable y de seis pozos con reservorio. A cada muestra de agua se le midió in situ el cloro residual mediante un comparador de cloro Hatch, método colorimétrico, usando para ello las pastillas DPD 1. En las muestras con cloro <0,05mg/L se realizó el cultivo según los manuales de procedimientos del Instituto Nacional de Salud (INS, Lima. Las cepas aisladas se enviaron al INS para confirmación diagnóstica y pruebas serológicas. Resultados: Se incluyeron 308 muestras de agua para consumo humano en ambos distritos (201 de pozos con bomba manuable y 107 con reservorio. Se realizó el aislamiento en 70(22,7% muestras: Aeromonas caviae 34(11,0%, Aeromonas hidrophyla 17(5,5% y Vibrio cholerae No O1 19(6,2%, no se encontró V. cholerae del serotipo O139. El Vibrio cholerae No O1 se aisló en 11(5,5% muestras de pozos con bomba manuable y en 8(7,4% pozos con reservorio, respectivamente. Conclusión: El agua de consumo humano proveniente de pozos tubulares representa un reservorio potencial para bacterias como Aeromonas y Vibrio cholerae, resaltando la necesidad de realizar la desinfección correspondiente de ésta antes de su consumo.

  17. Reporte histórico: Primer Aislamiento de Vibrio cholera serogrupo O1 biovar El Tor serovar Inaba durante la epidemia de cólera en el Perú ‑ 1991 Historical report: first isolation of Vibrio cholera serogroup O1 biovar El Tor serovar Inaba during the cholerae epidemic in Perú ‑ 1991

    Directory of Open Access Journals (Sweden)

    Nora Bravo Cruz

    2011-03-01

    Full Text Available Hace 20 años apareció una enfermedad diarreica nueva en el Perú y el Laboratorio de Referencia de Enteropatógenos del Instituto Nacional de Salud, cumplió una labor destacada en el aislamiento e identificación rápida y oportuna del Vibrio cholerae. La enfermedad del cólera no se había presentado anteriormente, pero en la última semana de enero de 1991 se detectó un brote epidémico de diarrea aguda con deshidratación intensa y algunos casos de fallecidos. La epidemia afectó, al comienzo, varias localidades del litoral peruano. Equipos de trabajo de la Oficina General de Epidemiología y de los laboratorios del Instituto Nacional de Salud obtuvieron muestras fecales de pacientes con diarrea aguda procedentes de las ciudades de Chancay, Chimbote, Piura y algunos hospitales de Lima. Las muestras colectadas en el medio de transporte de Cary y Blair fueron procesadas en el Laboratorio Nacional de Referencia de Enteropatógenos (LANARE del Instituto Nacional de Salud. De todas las muestras se aisló e identificó Vibrio cholerae serogrupo O1 biovar El Tor serovar Inaba que mostró ser sensible a la tetraciclina y a otros antibióticos. Esta investigación confirmó el primer brote epidémico de cólera en el Perú.20 years ago, a new diarrheal disease was introduced in Peru and the Enteropathogens Reference Laboratory of the Instituto Nacional de Salud had an outstanding role in the isolation and rapid and timely identification of Vibrio cholerae. Cholera had not been seen before, but during the last week of January 1991 an outbreak of acute diarrhea was detected, presenting intense dehydration and some deaths. The epidemic affected, in the beginning, many locations of the peruvian coast. Some working teams of the General Office of Epidemiology and of the Instituto Nacional de Salud obtained fecal samples from patients with acute diarrhea coming from the cities of Chancay, Chimbote, Piura and some hospitals in Lima. The collected samples

  18. Toxigenic Vibrio cholerae identified in estuaries of Tanzania using PCR techniques.

    Science.gov (United States)

    Dalusi, Lucy; Lyimo, Thomas J; Lugomela, Charles; Hosea, Ken M M; Sjöling, Sara

    2015-03-01

    The current study assessed the occurrence of the Vibrio cholerae serogroups O1 and O139 in environmental samples along salinity gradients in three selected estuaries of Tanzania both through culture independent methods and by cultured bacteria. Occurrence of V. cholerae was determined by PCR targeting the V. cholerae outer membrane protein gene ompW. Furthermore, the presence of toxigenic strains and serogroups O1 and O139 was determined using multiplex PCR with specific primers targeting the cholera toxin gene subunit A, ctxA, and serotype specific primers, O1-rfb and O139-rfb, respectively. Results showed that V. cholerae occurred in approximately 10% (n = 185) of both the environmental samples and isolated bacteria. Eight of the bacteria isolates (n = 43) were confirmed as serogroup O1 while one belonged to serogroup O139, the first reported identification of this epidemic strain in East African coastal waters. All samples identified as serogroup O1 or O139 and a number of non-O1/O139 strains were ctxA positive. This study provides in situ evidence of the presence of pathogenic V. cholerae O1 and O139 and a number of V. cholerae non-O1/O139 that carry the cholera toxin gene in estuaries along the coast of Tanzania.

  19. O1群霍乱弧菌胶体金法快速检测%Application of colloidal gold method in rapid detection of Vibrio cholerae O1

    Institute of Scientific and Technical Information of China (English)

    吴多荣; 周登仁; 李明刚; 李静粉; 韩小胜

    2009-01-01

    目的 了解胶体金法检测O1群霍乱弧菌的特异性、灵敏度和实用性.方法 取患者粪便标本,先用碱性蛋白胨水35℃增菌6 h,然后用胶体金法进行检测,同时作霍乱弧菌常规培养,培养出细菌后用血清凝集法作为鉴别诊断O1群霍乱弧菌的金标准,用统计学方法对胶体金法诊断试验的评价指标进行计算..结果 700份样本中,常规法培养出O1霍乱弧菌105株,胶体金法试验阳性108株,经χ2检验,差异无统计学意义(χ2=0.24,P>0.05),表明胶体金法检测O1群霍乱弧菌与常规培养法检测结果之间的差异无统计学意义.胶体金法检测O1群霍乱弧菌诊塑试验的灵敏度为93.33%,特异性为98.32%,阳性预测值为90.74%,阴性预测值为97.18%,假阴性率为1.68%,假阳性率为6.67%.结论 胶体金法检测O1群霍乱弧菌特异性高(98.32%),灵敏度好,与常规培养法的一致性也较高,在快速检测O1群霍乱弧菌中有一定应用价值.%Objective To understand the specificity, sensitivity and practicality of colloidal gold assay in the detection of Vibrio cholerae Ol group. Methods Stool specimens of patients were incubated in alkaline peptone water (35℃) for 6 hours. Then the colloidal gold method was used to detect Vibrio cholerae. At the same time conventional culture of Vibrio cholerae was identified with serum agglutination as the gold standard. The results of two methods were compared. Results Among 700 samples,105 Vibrio cholerae were identified with conventional methol method and 108 identified with coilidal gold assay and there was no significant difference between the two methods. (χ2 = 0.24 ,P > 0.05). The sensitivity of col-loidal gold method for Vibrio cholerae O1 detetion was 93.33% ,with a specificity of 98.32 ,a positive predictive value of 90. 74% ,a negative value of 97.18% ,a false negative rate of 1.68% ,and a false positive rate of 6.67%. Conclusion The colloidal gold method has good specificity and

  20. Antibacterial effect of Costus spiralis leaves extract on pathogenic strains of Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Celso Pérez

    2008-01-01

    pueden ser extraídos y purificados a partir de plantas para el desarrollo de nuevos medicamentos. Entre las diversas enfermedades que históricamente han afectado al hombre, el cólera ha sido potencialmente epidémico y una de las más sobresalientes. La bacteria Vibrio cholerae, el agente causal, puede ser eliminado mediante antibióticos, de modo que además del tratamiento tradicional de la enfermedad de rehidratación vía oral o intravenosa, comúnmente son aplicados antibióticos tales como la tetraciclina, ciprofloxacina, norfloxacina o azitromicina. El efecto antimicrobiano in vitro de extractos de hojas de Costus spiralis (Roscoe sobre varias cepas patógenas de Vibrio cholerae fue ensayado mediante la técnica de difusión en placas de agar. Hojas verdes de la plantas fueron colectadas, secadas en horno a 50 ºC durante 48 h, molidas y finalmente, sometidas a extracción con etanol. Luego de secado, el material residual fue resuspendido en agua destilada a 100 mg/mL (p/v y realizados los ensayos de actividad antimicrobiana. Aparentemente, las cepas patógenas que representan las pandemias del siglo xx: C7258 (O1, El Tor, Ogawa, C6706 (O1, El Tor, Inaba, O395 (O1, Clásica, Ogawa, CRC266 (O139 y 569B (O1, Clásica, Inaba fueron matadas, a juzgar por la presencia de halos de inhibición de crecimiento en los ensayos. Adicionalmente, se eterminaron las concentraciones mínimas inhibitorias de los extractos para las diferentes cepas. Los resultados anteriores fueron similares a los de la Ampicillina, lo que sugiere que Costus spiralis pude utilizarse como fuente de principios activos contra Vibrio cholerae.

  1. Low prevalence of Vibrio cholerae O1 versus moderate prevalence of intestinal parasites in food-handlers working with health care personnel in Haiti.

    Science.gov (United States)

    Llanes, Rafael; Somarriba, Lorenzo; Velázquez, Beltran; Núñez, Fidel A; Villafranca, Caridad M

    2016-01-01

    Food-handlers with poor personal hygiene working in food-service establishments could be potential sources of infection due to pathogenic organisms. In May 2011, a cross-sectional study was undertaken to determine the prevalence of bacteria and intestinal parasites among food-handlers working with Cuban health personnel in Haiti. Stool specimens were collected from 56 food-handlers and samples were examined using standard procedures. Of the food handlers, 26.8% had one bacterial or intestinal parasite. The most prevalent species of organism found were Blastocystis spp. (9%), followed by Vibrio cholerae O1 serotype Ogawa, Aeromonas spp. and Giardia intestinalis, each one with 4%. The prevalence of intestinal parasites was 19.7%. Five out of 56 food handlers had diarrhea at the time the study was conducted. It was found that there was a lower prevalence of V. cholerae O1 serotype Ogawa in comparison to intestinal parasites. The study highlights the importance of the precautions that must be taken in cholera-affected countries by medical teams and their organizations, with emphasis on the preparation, processing, and serving of meals. The recommendation is to intensify continuing education programs, periodical laboratory examinations to detect carriers and food-handlers reporting sick, and to observe strict adherence to hygienic food-handling practices. In addition, food handlers with diarrhea should refrain from preparation or delivery of food.

  2. Antimicrobial resistance of clinical and environmental strains of Vibrio cholerae isolated in Lima-Peru during epidemics of 1991 and 1998

    Directory of Open Access Journals (Sweden)

    J.O. Ibarra

    2007-02-01

    Full Text Available The susceptibility in vitro of 71 isolations of V. cholerae was evaluated: 24 of clinical origin and 47 strains of clinical and environmental origin collected in the epidemic of 1991 and during the outbreak epidemic of 1998 in Lima-Peru respectively. The biochemical and serological tests carried out established that 43 (60,6% corresponded to the serogroup O1 Ogawa of the 1998 epidemic; 26 (36.6% were of the serotype Inaba, being 24 of them isolated in 1991. Two strains did not belong to the serogroup O1. By means of disk diffusion method and Minimal Inhibitory Concentration (MIC, 15 strains with multi-resistance to antibiotics were determined, 10 of which were of clinical origin and 5 of natural origin, showing 9 antibiotypes with different resistance pattern. The evaluation of susceptibility in front of the vibriostatic agent O/129, demonstrated that 11.4% of the strains, collected in 1998, presented resistance to a concentration of 150 µg. A direct relationship among the resistance that presented the strains of clinical and environmental origin isolated in 1991 and 1998 was established as much for tetracycline, sulfa/trimethoprim and 0/129; 88.6% of the clinical strains of the year 1998 presented resistance to these three drugs, while 100% of clinical strains isolated in 1991 were sensitive to O/129 (150 µg, sulfa/trimethoprim and tetracycline. We conclude that V. cholerae O1 has increased its resistance to antimicrobial drugs of clinical use in the same way it is also losing susceptibility to the vibriostatic compound O/129 for what their use is not recommended for taxonomic purposes.

  3. IncA/C plasmids harboured in serious multidrug-resistant Vibrio cholerae serogroup O139 strains in China.

    Science.gov (United States)

    Wang, Ruibai; Yu, Dong; Zhu, Lianhui; Li, Jie; Yue, Junjie; Kan, Biao

    2015-03-01

    Vibrio cholerae serogroup O139 emerged in 1992 and is one of two major serogroups to have caused cholera epidemics. After 1998, serious multidrug-resistant (MDR) O139 strains quickly became common in China, showing a multidrug resistance profile to eight antibiotics. It is a great threat to public health, and elucidation of its mechanisms of resistance will provide a helpful guide for the clinical treatment and prevention of cholera. In this study, mega-plasmids from MDR V. cholerae O139 strains were identified by pulsed-field gel electrophoresis (PFGE) without enzyme digestion. One plasmid was isolated and sequenced, belonging to the IncA/C family. Ten antibiotic resistance genes were found in the MDR regions, including a blaTEM-20 gene, and these genes endowed the host with resistance to seven antibiotics. This kind of plasmid was positive in 71.2% (198/278) of toxigenic O139 strains, and the rate of plasmid positivity was consistent with the yearly change in MDR rates of these strains. This study reveals an important role of the IncA/C family plasmid in the spread of multiple antibiotic resistance of epidemic V. cholerae serogroup O139 strains, which has recombined with plasmids from different bacterial species and transferred among V. cholerae strains.

  4. Vibrio cholerae O1 El Tor from southern Vietnam in 2010 was molecularly distinct from that present from 1999 to 2004.

    Science.gov (United States)

    Nguyen, V H; Pham, H T; Diep, T T; Phan, C D H; Nguyen, T Q; Nguyen, N T N; Ngo, T C; Nguyen, T V; Do, Q K; Phan, H C; Nguyen, B M; Ehara, M; Ohnishi, M; Yamashiro, T; Nguyen, L T P; Izumiya, H

    2016-04-01

    The Vibrio cholerae O1 (VCO1) El Tor biotype appeared during the seventh cholera pandemic starting in 1961, and new variants of this biotype have been identified since the early 1990s. This pandemic has affected Vietnam, and a large outbreak was reported in southern Vietnam in 2010. Pulsed-field gel electrophoresis (PFGE) and multilocus variable-number tandem-repeat analyses (MLVA) were used to screen 34 VCO1 isolates from the southern Vietnam 2010 outbreak (23 patients, five contact persons, and six environmental isolates) to determine if it was genetically distinct from 18 isolates from outbreaks in southern Vietnam from 1999 to 2004, and two isolates from northern Vietnam (2008). Twenty-seven MLVA types and seven PFGE patterns were identified. Both analyses showed that the 2008 and 2010 isolates were distinctly clustered and separated from the 1999-2004 isolates.

  5. Structural organization of the transfer RNA operon I of Vibrio cholerae: Differences between classical and El Tor strains

    Indian Academy of Sciences (India)

    Atreyi Ghatak; Anasuya Majumdar; Ranajit K Ghosh

    2005-09-01

    Nine major transfer RNA (tRNA) gene clusters were analysed in various Vibrio cholerae strains. Of these, only the tRNA operon I was found to differ significantly in V. cholerae classical (sixth pandemic) and El Tor (seventh pandemic) strains. Amongst the sixteen tRNA genes contained in this operon, genes for tRNA Gln3 (CAA) and tRNA Leu6 (CUA) were absent in classical strains as compared to El Tor strains. The observation strongly supported the view that the above two pandemic strains constitute two different clones.

  6. 浙江省非O1、非O139群霍乱弧菌快速检测与毒力基因研究%Rapid detection of non O1 and non O139 Vibrio cholerae and their virulence genes in Zhejiang province

    Institute of Scientific and Technical Information of China (English)

    罗芸; 叶菊莲; 金大智; 张政; 王复甦; 徐宝祥

    2011-01-01

    目的 建立一种快速检测非O1、非O139群霍乱弧菌方法,并对浙江省部分地区海水产品中非O1、非O139 群霍乱弧菌及其携带毒力基因开展快速检测.方法 采用三糖斜面、氧化酶实验、粘丝实验、无盐胨水等简单生化进行初筛,对全部结果阳性的菌株用聚合酶链反应(PCR)方法检测霍乱弧菌外膜蛋白OmpW以及毒力基因ctxA、hlyA和toxR.结果 按照方法用三糖斜面初筛后共有504份样本阳性,其中氧化酶、粘丝和无盐胨水全部为阳性的有434份;有388份样本OmpW PCR结果为阳性,其中ctxA结果均为阴性,toxR阳性hlyA阳性的有225株.结论 建立的快速检测方法提高了非O1、非O139群霍乱弧菌的检出率,缩短了检测时间,节约了检测成本.浙江省非O1、非O139群霍乱弧菌在海水产品中广泛存在,应对其致病性提高警惕,加强监测.%Objective To establish a rapid detection assay to detect non O1 and non-O139 Vibrio cholerae in sea foods and their virulence genes in Zhejiang province. Methods Non Ol and non O139 Vibrio cholerae strains were detected by using triple sugar tube, oxidase test, ropiness test and saltless peptone water test, then OmpW,ctxA、hlyA and toxR genes of Vibrio cholerae were detected by PCR assay. Results Totally 504 sample were positive by using triple sugar tube, in addition, 434 samples were positive in other 3 tests. Totally 388 strains were OmpW gene positive, but ctxA gene negative and 225 strains were both hlyA and toxR gene positive. Conclusion The established rapid assay could increase the detection rate of non Ol and non O139 Vibrio cholerae, shorten the detection time and reduce test cost. Non Ol and non-O139 Vibrio cholerae generally exist in sea foods in Zhejiang province. It is important to fully understand their pathogenicities and strengthen the disease surveillance.

  7. Obtención de extractos de membrana externa de Vibrio cholerae O1, mediante el uso de diferentes detergentes

    Directory of Open Access Journals (Sweden)

    José Luis Pérez

    2006-04-01

    Full Text Available En la actualidad existen dos variantes principales de vacunas orales contra el cólera: una basada en células inactivadas de diferentes biotipos y serotipos y otra basada en la administración de cepas vivas genéticamente atenuadas. Una vacuna por subunidades pudiera ser una variante muy atractiva. Este trabajo describe la purificación parcial y caracterización preliminar de extractos de proteínas de membrana externa-lipopolisacárido (PME-LPS, obtenidos a partir de Vibrio cholerae O1, con el interés de seleccionar un proteoliposoma que posteriormente será estructurado en forma de cocleatos para su uso por vía oral en humanos. Las preparaciones fueron obtenidas a través del uso de diferentes detergentes. La cantidad de LPS en cada preparación fue estimada mediante la determinación de las unidades endotóxicas en el ensayo del Limulus (LAL. La composición de cada muestra fue evaluada mediante SDS-PAGE y Dot Blot. La inoculación intranasal (IN en ratones Balb/c se utilizó para la evaluación de la inmunogenicidad de las preparaciones, y la respuesta inmune fue determinada por ELISA y el título de anticuerpos vibriocidas. El tamaño molecular de la preparación con mejores resultados en inmunogenicidad se estimó mediante la cromatografía en Sephacryl S-1000. Se obtuvieron diferentes perfiles electroforéticos de acuerdo con el tipo de detergente utilizado. El LPS fue identificado en todas las preparaciones y aquella obtenida con el SDS al 15% mostró la más baja relación proteínas/LPS y los mejores resultados en los ensayos de inmunogenicidad. Adicionalmente se comprobó que su tamaño molecular es similar al observado en el proteoliposoma de VAMENGOC- BC. La preparación obtenida con el SDS al 15% constituye un proteoliposoma, con capacidad para estimular altos niveles de anticuerpos IgG anti-LPS y altos títulos de anticuerpos vibriocidas, luego de su administración por vía intranasal en ratones. Estos resultados constituyen

  8. Misidentification of Vibrio cholerae O155 isolated from imported shrimp as O serogroup O139 due to cross-agglutination with commercial O139 antisera

    DEFF Research Database (Denmark)

    Dalsgaard, A.; Mazur, J.; Dalsgaard, Inger

    2002-01-01

    Fish and shellfish products imported into Denmark are routinely analyzed for pathogenic Vibrio spp., particularly Vibrio cholerae, if products originate from subtropical or tropical areas. A V. cholerae strain that agglutinated commercial O139 antiserum but not the O1, Inaba, or Ogawa antisera...... was isolated from imported raw frozen shrimp. The toxigenicity of the strain was analyzed, and the results of a polymerase chain reaction showed that the V. cholerae strain did not contain the virulence genes ctx, tcp9, and zot, which are normally found in V. cholerae O1 and O139. The strain was resistant...

  9. Comparative phenotypic characterization of Vibrio cholerae isolates collected from aquatic environments of Georgia.

    Science.gov (United States)

    Kokashvili, T; Elbakidze, T; Jaiani, E; Janelidze, N; Kamkamidze, G; Whitehouse, C; Huq, A; Tediashvili, M

    2013-11-01

    Vibrio cholerae is ubiquitous in aquatic environment inhabiting marine, fresh and brackish waters. V. cholerae serotypes O1 and O139 cause the devastating diarrheal disease cholera, which is often fatal without proper treatment. Little is known regarding the abundance and diversity of clinically important nonhalophilic vibrios in the South Caucasus region, particularly in Georgia. Here we provide the data on the Georgian environmental strains of V. cholerae isolated in 2006-2009 years from the coastal waters of the Black Sea and inland water reservoirs near Tbilisi. In total, 846 V. cholerae strains were collected from the water samples, most of them (705 strains) obtained from fresh water lakes. Isolation pattern of V. cholerae showed obvious seasonality with the highest isolation rates in late summer - early autumn. Twenty-nine isolates of V. cholerae were attributed to the O1 serotype based on serological studies and PCR identification and were further grouped by biochemical properties into classical and El Tor biotypes as well as hybrids. The study of antibiotic susceptibility profiles for V. cholerae isolates showed that 95% were sensitive to tetracycline, 91% to doxycycline, and 91% to ciprofloxacin. Interestingly, the freshwater isolates appeared to be more resistant to antibiotics than the Black Sea isolates. Among Black Sea isolates of V. cholerae toxigenic strains of O1 serotype revealed higher antibiotic resistance compared to non- O1/non-O139 isolates. In addition, V. cholerae O1 and non- O1/non-O139 isolates differed by phage susceptibility profiles, with higher diversity within the population of environmental non-O1/non-O139 V. cholerae isolates.

  10. Purification and partial characterization of a non-O1 Vibrio cholerae hemolysin that cross-reacts with thermostable direct hemolysin of Vibrio parahaemolyticus.

    OpenAIRE

    Yoh, M; Honda, T.; Miwatani, T

    1986-01-01

    A newly identified hemolysin (NAG-rTDH), which is related to the thermostable direct hemolysin (Vp-TDH) of Vibrio parahaemolyticus produced by non-O1 Vibrio cholerae, was studied. NAG-rTDH was purified by successive column chromatographies on DEAE-cellulose and an immunoaffinity column coupled with anti-Vp-TDH immunoglobulin. The molecular weight of NAG-rTDH was estimated as 18,500, similar to that of Vp-TDH, as judged by sodium dodecyl sulfate slab gel electrophoresis, but its charge or mole...

  11. Development of Gold-immunochromatography Test for Rapid Detection of Vibrio Cholerae Group O1%霍乱弧菌O1胶体金免疫层析快速检测法的建立

    Institute of Scientific and Technical Information of China (English)

    王玉金; 杨书豪; 刘丽; 庞向宇; 孙晨阳; 李珊珊

    2013-01-01

    Objective To develop a rapid and simple colloidal gold immunochromatographic ssay ( GICA) for the detection of Vibrio cholerae group 01. Methods A colloidal gold immunochromatography test based on double-antibody sandwich assay for detecting Vibrio cholerae group 01 was developed. Its sensitivity, specificity and stability were then evaluated. A total of 208 clinical specimens were determined with the prepared GICA, and the results were compared with those by bacterial culture. Results Typical detecting time was less than 10 minutes per sample. The positive samples produced a positive signal on immunochromatographic strip, whereas the negative one did not. The sensitivity of the test was 1 x 105 cfu/ml. Both the specificity and sensitivity of the prepared GICA in determination of 208 clinical specimens were 100%. Conclusion The gold-immunochromatography test appears to be a rapid, convenient, specific and sensitive test for detecting Vibrio cholerae group 01 on site.%目的 建立一种快速、简易的检测霍乱弧菌O1群的胶体金免疫层析法(GICA).方法 利用胶体金免疫层析技术,采用双抗体夹心法检测霍乱弧菌O1群,对该法进行敏感性、特异性和稳定性分析.与细菌培养法对比检测208份临床标本.结果 该法能在10分钟内完成检测;该试纸条仅与霍乱弧菌O1群阳性样品发生特异性反应;检测霍乱弧菌O1群的最低检出浓度为1×105cfu/mL;与细菌培养法对比检测208份临床标本,特异性和灵敏度均达100%.结论 新建立的霍乱弧菌O1群胶体金免疫层析试验简便、快速,特异性和灵敏度较好,适用于现场样品的快速筛查.

  12. Cholera outbreaks in the classical biotype era.

    Science.gov (United States)

    Siddique, A K; Cash, Richard

    2014-01-01

    In the Indian subcontinent description of a disease resembling cholera has been mentioned in Sushruta Samita, estimated to have been written between ~400 and 500 BC. It is however not clear whether the disease known today as cholera caused by Vibrio cholerae Vibrio cholerae O1 is the evolutionary progression of the ancient disease. The modern history of cholera began in 1817 when an explosive epidemic broke out in the Ganges River Delta region of Bengal. This was the first of the seven recorded cholera pandemics cholera pandemics that affected nearly the entire world and caused hundreds of thousands of deaths. The bacterium responsible for this human disease was first recognised during the fifth pandemic and was named V. cholerae which was grouped as O1, and was further differentiated into Classical and El Tor biotypes. It is now known that the fifth and the sixth pandemics were caused by the V. cholerae O1 of the Classical biotype Classical biotype and the seventh by the El Tor biotype El Tor biotype . The El Tor biotype of V. cholerae, which originated in Indonesia Indonesia and shortly thereafter began to spread in the early 1960s. Within the span of 50 years the El Tor biotype had invaded nearly the entire world, completely displacing the Classical biotype from all the countries except Bangladesh. What prompted the earlier pandemics to begin is not clearly understood, nor do we know how and why they ended. The success of the seventh pandemic clone over the pre-existing sixth pandemic strain remains largely an unsolved mystery. Why classical biotype eventually disappeared from the world remains to be explained. For nearly three decades (1963-1991) during the Seventh cholera pandemic seventh pandemic, cholera in Bangladesh has recorded a unique history of co-existence of Classical and El Tor biotypes of V. cholerae O1 as epidemic and endemic strain. This long co-existence has provided us with great opportunity to improve our understanding of the disease itself

  13. Cuantificación de lipopolisacárido en un proteoliposoma obtenido a partir de la superficie externa de Vibrio cholerae O1

    Directory of Open Access Journals (Sweden)

    José Luis Pérez

    2007-04-01

    Full Text Available El lipopolisacárido (LPS de Vibrio cholerae O1, a diferencia de la mayoría de las bacterias gramnegativas, no es reactivo en el ensayo del KDO, un método ampliamente utilizado para la cuantificación de LPS. Este LPS puede ser cuantificado por peso seco cuando se trata del antígeno en estado puro, pero cuando se necesita cuantificarlo, por ejemplo en un proteoliposoma, se requieren métodos de hidrólisis fuerte para que la molécula de KDO se exponga y sea reactiva en el ensayo antes mencionado. En este trabajo describimos un método rápido, sencillo y reproducible que nos permite cuantificar el LPS en un proteoliposoma obtenido a partir de la superficie externa de V. cholerae O1, El Tor Ogawa, mediante el uso de un anticuerpo monoclonal anti-LPS Ogawa en Western blot y la posterior evaluación del perfil reactivo en un densitómetro, utilizando LPS Ogawa purificado y cuantificado por peso seco como patrón de referencia. Este método puede ser aplicado a cualquier otra preparación que contenga LPS y que no pueda ser evaluada por KDO.

  14. Identification and virulence gene detection of non-O1 and non-O139 Vibrio chol-erae isolates causing septicemia%致血流感染非 O1群非 O139群霍乱弧菌的鉴定及毒力基因检测

    Institute of Scientific and Technical Information of China (English)

    邹玖明; 张爱平; 李智山; 杨燕; 赵建忠

    2014-01-01

    Objective To identify an suspected Vibrio cholerae isolated from Xiangyang Central Hospital and characterize the strain in terms of antibiotic resistance and relevant virulence genes.Methods Pathogen identification and susceptibility testing were completed with MicroScan WalkAway 40 Automated Microbiology System.Slide agglutination was used for serotyping. PCR and sequencing technology were employed for 16s RNA gene analysis.PCR technique was used to detect six major viru-lence genes.Results This suspectedVibrio cholerae isolate was confirmed as non-O1 and non-O139 Vibrio cholerae .Suscep-tibility testing results indicated that the strain was sensitive to ampicillin,chloramphenicol,trimethoprim-sulfamethoxazole, and tetracycline.16s RNA gene sequence analysis showed 100% homologous with the registered sequence in National Center for Biotechnology Information database.Virulence genes rtxC and toxR were identified.The other virulence genes such as tcpAET,ctxA,hlyA,and tcpACL were negative.Conclusions This suspected Vibrio cholerae isolate is confirmed as non-O1 and non-O139 Vibrio cholerae .The pathogenic factors may be related to the virulence genes rtxC and toxR.%目的:对襄阳市中心医院分离1株疑似霍乱弧菌进行鉴定及药物敏感性试验,并检测其主要毒力基因。方法利用MicroScan WalkAway 40鉴定仪进行生化鉴定及药物敏感性试验,玻片凝集法确定其血清型别,应用 PCR 及测序技术分析其16SrRNA 基因;PCR 检测其6个毒力基因。结果经鉴定,该株疑似霍乱菌株为非 O1群非 O139群霍乱弧菌,经16SrRNA分析与美国国家生物技术信息中心数据库中霍乱弧菌相似性达100%。药敏试验结果显示该菌对氨苄西林、氯霉素、甲氧苄啶-磺胺甲口恶唑、四环素均敏感,毒力基因检测 rtxC 和 toxR 阳性,tcpAET、ctx A 、hlyA、tcpACL 阴性。结论该株疑似霍乱弧菌为非 O1群非 O139群霍乱弧菌,其致病与 rtxC、toxR 毒力基因有关。

  15. Characterization and restriction analysis of the P sex factor and the cryptic plasmid of Vibrio cholerae strain V58.

    Science.gov (United States)

    Bartowsky, E J; Morelli, G; Kamke, M; Manning, P A

    1987-07-01

    The P plasmid of Vibrio cholerae is a derepressed sex factor restricted to V. cholerae and has been shown to express surface exclusion. We have isolated the plasmids of strain V58 and have found that in addition to P, two further cryptic plasmids are also present. P has a size of 68 kb as determined by both electron microscopy and restriction endonuclease analysis. These other plasmids are 34 and 4.7 kb in size. Restriction maps of P and the larger cryptic plasmid have been determined. It has been demonstrated that P differs from the standard Inc group test plasmids and also expresses a surface exclusion system. The ability of the type Inc plasmids to be transferred to V. cholerae by either liquid or filter matings and the stability of these plasmids in V. cholerae have also been examined.

  16. Molecular analysis on non-O1 and non-O139 Vibrio cholerae isolates%非O1/非O139群霍乱弧菌感染病例分离株分子流行病学分析

    Institute of Scientific and Technical Information of China (English)

    陈道利; 胡万富; 詹圣伟; 景怀琦; 阚飙; 张萍; 王多春; 陈谨; 喻佰启; 程险峰; 刁保卫; 周海健; 朱明

    2012-01-01

    目的分析安徽省马鞍山市连续2个月内监测到的6例具有霍乱疑似症状、感染非O1/非O139群霍乱弧菌的病例,判断疫情的聚集性.方法 对病例分离株进行生化和血清型别鉴定以及溶血试验,药敏试验检测抗生素耐药谱,应用荧光PCR和常规PCR进行霍乱弧菌特异基因、毒力及其相关基因的检测,包括ompW、ctx 、tcpA、toxR、hlyA 、zot、ace、rstR和gⅢCTX,应用脉冲场凝胶电泳(PFGE)分析其分子型别.结果 生化鉴定和血清学试验鉴别腹泻病例6株菌株为非O1/非O139霍乱弧菌,均产生β溶血;14种药物中有12种属于全部敏感;荧光PCR检测霍乱弧菌特异性基因ompW均为阳性,ctx、tcpA 、zot、ace、rstR和gⅢCTX基因均为阴性,toxR 、hlyA基因有5株菌扩增阳性,1株菌(1001434446)为阴性;PFGE显示6株菌带型均不相同,但有2株非常相似,分离株与霍乱弧菌产毒株相似性很低.结论 6例感染非产毒的非O1/非O139群霍乱弧菌病例发病虽相对集中,但属于散发病例,在局部地区频繁出现,提示其公共卫生意义不可忽视.%Objective According to results from the two-month consecutive surveillance program in Maanshan,six suspected cases of non-O1 non-O139 Vibrio (V.) cholerae infection,were found that called for identification of pathogens as well as molecular-epidemiological analysis to determine the aggregation of the epidemic situation.Methods Biochemical and serotype identification,hemolysis test,and drug sensitive test were used to detect the drug resistance spectrum.Real-time PCR and conventional PCR were used to detect the presence of V.cholerae specific genes,virulent genes and its related genes,including ompW,ctx,tcpA,toxR,hlyA,zot,ace,rstR and g ⅢCTX.Pulsed-field gel electrophoresis (PFGE) was used to analyze the molecular type of strains.Results All the six isolates of non-O 1 non-O 139 V.cholerae were identified by biochemical and serologic tests,and appeared to be

  17. Molecular characteristics and antibiotic resistances of Vibrio cholerae O1 isolates in Hangzhou in 2009%2009年杭州市01群霍乱弧菌的分子特征及耐药性

    Institute of Scientific and Technical Information of China (English)

    郑伟; 俞骅; 汪皓秋; 张蔚; 潘劲草

    2011-01-01

    PFGE patterns ( P1 - P11 ). Twenty-three isolates with genotype ctxA - and tcpA + were clustered into 7 PFGE patterns( P1 - P7 ,termed P1-like cluster) with the similarity to be equal or greater than 91.4% ,and 56. 52% (13/23) of them belong to P1. 7 isolates with very high similarity (97.6%), belonging to P1 (6 isolates),and P2 (1 isolate), respectively, were collected from one foodborne disease outbreak. The resistant rates of the 24 isolates with genotype ctxA - and tcpA + to ampicillin, tobramicin and amikacin were 20. 83 % ( 5/24 ), 4. 17 % ( 1/24 ) and 4. 17 % ( 1/24 ), respectively. Conclusion The genotype of the epidemic strains of V. cholerae O1 isolates in Hangzhou in 2009 with high similarity was ctxA - and tcpA + ;The level of drug resistances of this kind of V. cholerae O1 isolates were not high.

  18. Nitrosative Stress Response in Vibrio cholerae: Role of S-Nitrosoglutathione Reductase.

    Science.gov (United States)

    Patra, Sourav Kumar; Bag, Prasanta Kumar; Ghosh, Sanjay

    2016-12-20

    Vibrio cholerae, the causative agent of cholera, poses serious threats to humans worldwide. V. cholerae faces host inflammatory response and encounters nitrosative stress before establishing successful colonization. It is not clear how V. cholerae combats nitric oxide and reactive nitrogen species. In the present study, we used three clinical strains of V. cholerae and tested their nitrosative stress response pattern towards sodium nitroprusside (SNP) and S-Nitrosoglutathione (GSNO). Among them, V. cholerae, belonging to both O1 and O139 serotypes, showed moderate resistance to SNP and GSNO. However, a V. cholerae strain belonging to non O1 and non O139 showed sensitivity to SNP but resistance towards GSNO. Reduced glutathione and glutathione reductase play a significant role to combat nitrosative stress in V. cholerae. This is the first report where we show the presence of GSNO reductase activity in V. cholerae and that it plays an important role to detoxify S-Nitrosoglutathione. GSNO reductase activity of V. cholerae was regulated by posttranslational modification through S-nitrosylation under in vitro conditions which could be reversed by dithiothreitol (DTT). In addition, we show that biofilm formation remained unaffected under nitrosative stress in V. cholerae.

  19. The Zymovars of Vibrio cholerae: Multilocus Enzyme Electrophoresis of Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Fernanda S Freitas

    2002-06-01

    Full Text Available Zymovars analysis also known as multilocus enzyme electrophoresis is applied here to investigate the genetic variation of Vibrio cholerae strains and characterise strains or group of strains of medical and epidemiological interest. Fourteen loci were analyzed in 171 strains of non-O1 non-O139, 32 classical and 61 El Tor from America, Africa, Europe and Asia. The mean genetic diversity was 0.339. It is shown that the same O antigen (both O1 and non-O1 may be present in several geneticaly diverse (different zymovars strains. Conversely the same zymovar may contain more than one serogroup. It is confirmed that the South American epidemic strain differs from the 7th pandemic El Tor strain in locus LAP (leucyl leucyl aminopeptidase. Here it is shown that this rare allele is present in 1 V. mimicus and 4 non-O1 V. cholerae. Non toxigenic O1 strains from South India epidemic share zymovar 14A with the epidemic El Tor from the 7th pandemic, while another group have diverse zymovars. The sucrose negative epidemic strains isolated in French Guiana and Brazil have the same zymovar of the current American epidemic V. cholerae.

  20. Characterization and genetic variation of vibrio cholerae isolated from clinical and environmental sources in Thailand

    DEFF Research Database (Denmark)

    Siriphap, Achiraya; Leekitcharoenphon, Pimlapas; Kaas, Rolf Sommer

    2017-01-01

    Cholera is still an important public health problem in several countries, including Thailand. In this study, a collection of clinical and environmental V. cholerae serogroup O1, O139, and non-O1/non-O139 strains originating from Thailand (1983 to 2013) was characterized to determine phenotypic...... and genotypic traits and to investigate the genetic relatedness. Using a combination of conventional methods and whole genome sequencing (WGS), 78 V. cholerae strains were identified. WGS was used to determine the serogroup, biotype, virulence, mobile genetic elements, and antimicrobial resistance genes using...... online bioinformatics tools. In addition, phenotypic antimicrobial resistance was determined by the minimal inhibitory concentration (MIC) test. The 78 V. cholerae strains belonged to the following serogroups O1: (n = 44), O139 (n = 16) and non-O1/non-O139 (n = 18). Interestingly, we found...

  1. Antibiotic resistance of Vibrio cholera strains isolated from imported crocodiles from Thailand%泰国进口鳄鱼霍乱弧茵药敏试验

    Institute of Scientific and Technical Information of China (English)

    陈冬娥; 陈冠武; 苏建晖; 许先凯

    2012-01-01

    Twelve Vibrio cholera strains were isolated from sixty anal swabs in sampling inspection of the Crocodiles imported from Thailand. It was confirmed that the strains were non-O1, non-O139 Vibrio choleras by biochemical test and serological identification. Eighteen drugs were used in the antibiotic resistance test. The test showed that all the strains were sensitive to most drugs, moderately susceptible to acheomycin and resistance to ampicillin and piperacillin.%在某鳄鱼养殖场抽检的60份泰国进口鳄鱼肛拭子样品中,有12份检出霍乱弧菌。通过生化试验和血清学分型等鉴定,确认12株霍乱弧菌均为非0。非0。。群霍乱弧菌。采用18种抗菌药物进行药敏试验,结果显示所有菌株对大多数抗菌药物高度敏感,对四环素等药物中度敏感,对氨苄青霉素和氧呱嗪青霉素不敏感。

  2. An Electrochemical Strategy using Multifunctional Nanoconjugates for Efficient Simultaneous Detection of Escherichia coli O157: H7 and Vibrio cholerae O1

    Science.gov (United States)

    Li, Yan; Xiong, Ya; Fang, Lichao; Jiang, Lili; Huang, Hui; Deng, Jun; Liang, Wenbin; Zheng, Junsong

    2017-01-01

    The rapid and accurate quantification of the pathogenic bacteria is extremely critical to decrease the bacterial infections in all areas related to health and safety. We have developed an electrochemical strategy for simultaneous ultrasensitive detection of E. coli O157:H7 and Vibrio cholerae O1. This approach was based on the specific immune recognition of different pathogenic bacteria by multifunctional nanoconjugates and subsequent signal amplification. By employing the proposed biosensor, the concentrations of these pathogenic bacteria could be established on a single interface in a single run with improved sensitivity and accuracy. The successful approach of the simultaneous detection and quantification of two bacteria by an electrochemical biosensor demonstrated here could be readily expanded for the estimation of a variety of other pathogenic bacteria, proteins, and nucleotides. Because of their high sensitivity, electrochemical biosensors may represent a new avenue for early diagnosis of diseases. PMID:28382165

  3. Acanthamoeba polyphaga is a possible host for Vibrio cholerae in aquatic environments.

    Science.gov (United States)

    Sandström, Gunnar; Saeed, Amir; Abd, Hadi

    2010-09-01

    Acanthamoeba is a genus of free-living amoebae found to be able to host many bacterial species living in the environment. Acanthamoebae and Vibrio cholerae are found in the aquatic environments of cholera endemic areas. Previously it has been shown that V. cholerae O1 and O139 can survive and grow in Acanthamoeba castellanii. The aim of this study was to examine the ability of Acanthamoeba polyphaga to host V. cholerae O1 and O139. The interaction between A. polyphaga and V. cholerae strains was studied by means of viable amoeba cell counts and viable count of the bacteria in the absence and presence of amoebae. The viable count of intracellularly growing bacteria was estimated by utilizing gentamicin assay. Electron microscopy was used to determine the localization of V. cholerae inside A. polyphaga. The results showed that A. polyphaga enhanced growth and survival of V. cholerae, which grew and survived inside the amoeba cells for 2weeks. The electron microscopy showed that A. polyphaga hosted intracellular V. cholerae localized in the vacuoles of amoeba cell. Neither the presence of V. cholerae together with A. polyphaga nor the intracellular localization of the bacteria inhibited growth and survival of A. polyphaga. The outcome of the interaction between these microorganisms may support strongly the role of A. polyphaga as host for V. cholerae O1 and O139.

  4. Environmental surveillance for toxigenic Vibrio cholerae in surface waters of Haiti.

    Science.gov (United States)

    Kahler, Amy M; Haley, Bradd J; Chen, Arlene; Mull, Bonnie J; Tarr, Cheryl L; Turnsek, Maryann; Katz, Lee S; Humphrys, Michael S; Derado, Gordana; Freeman, Nicole; Boncy, Jacques; Colwell, Rita R; Huq, Anwar; Hill, Vincent R

    2015-01-01

    Epidemic cholera was reported in Haiti in 2010, with no information available on the occurrence or geographic distribution of toxigenic Vibrio cholerae in Haitian waters. In a series of field visits conducted in Haiti between 2011 and 2013, water and plankton samples were collected at 19 sites. Vibrio cholerae was detected using culture, polymerase chain reaction, and direct viable count methods (DFA-DVC). Cholera toxin genes were detected by polymerase chain reaction in broth enrichments of samples collected in all visits except March 2012. Toxigenic V. cholerae was isolated from river water in 2011 and 2013. Whole genome sequencing revealed that these isolates were a match to the outbreak strain. The DFA-DVC tests were positive for V. cholerae O1 in plankton samples collected from multiple sites. Results of this survey show that toxigenic V. cholerae could be recovered from surface waters in Haiti more than 2 years after the onset of the epidemic.

  5. 异育银鲫非O1/非O139群霍乱弧菌的分离及鉴定%Isolation and identification of non-O1/non-O139 Vibrio cholerae from gibel carp

    Institute of Scientific and Technical Information of China (English)

    秦蕾; 徐静; 张晓君

    2013-01-01

    为鉴定引起异育银鲫发病的疫情的病原,本研究从病鱼体内分离到细菌分离株,经动物回归试验证明为导致异育银鲫发病的致病原.对病原菌株进行形态学、生理生化测定以及16S rDNA序列分析,显示该菌株与霍乱弧菌(Vibrio cholerae)表型特征一致,而且在系统发育树中也是与霍乱弧菌相聚类,两者16S rDNA序列相似性高达99.93%.综合表型特征与基因序列分析的结果表明,该病原菌株应归属于霍乱弧菌.血清型鉴定为非O1/非O139群霍乱弧菌.药物敏感性试验结果显示该株霍乱弧菌对喹诺酮类、头孢类、氨基糖苷类、大环内酯类,四环素类以及β-内酰胺类等药物敏感.%An infectious disease occurred in a gibel carp farm in 2010. The dominant bacteria were isolated from diseased fish and proved to be causal agent of the disease by experimental infection in gibel carp. Conventional physiological and biochemical tests showed that the isolates had phenotypic characteristics in according with reference strain of Vibrio cholerae. And 16S rDNA sequence was analyzed revealing that it was most closely related to V.cholerae with 99.93% similarity. Therefore, the isolates were finally identified as V.cholerae. Antibiotic susceptibility testing showed that the isolates were sensitive to quinolones, cephems, aminoglycosides, macrolide, tetracyclines and β-lactams.

  6. Neutrophils Are Essential for Containment of Vibrio cholerae to the Intestine during the Proinflammatory Phase of Infection

    OpenAIRE

    Queen, Jessica; Satchell, Karla J Fullner

    2012-01-01

    Cholera is classically considered a noninflammatory diarrheal disease, in comparison to invasive enteric organisms, although there is a low-level proinflammatory response during early infection with Vibrio cholerae and a strong proinflammatory reaction to live attenuated vaccine strains. Using an adult mouse intestinal infection model, this study examines the contribution of neutrophils to host defense to infection. Nontoxigenic El Tor O1 V. cholerae infection is characterized by the upregula...

  7. 口服重组B亚单位O1/O139霍乱疫苗的制备及检定%Preparation and quality control of oral bivalent recombinant cholera toxin B subunit-O1/O139 whole cell cholera vaccine

    Institute of Scientific and Technical Information of China (English)

    张静飞; 陈磊; 董思国; 曹天成; 何金娜; 曹欣; 胡鹏; 郝倩; 魏文进

    2016-01-01

    目的 制备口服重组B亚单位O1/O139霍乱疫苗,并进行全面检定.方法 制备O1、O139群霍乱弧菌灭活菌体原液及重组霍乱毒素B亚单位(recombinant cholera toxin B subunit,rCTB)原液,按比例混合制成口服重组B亚单位O1/O139霍乱疫苗,为保护rCTB免受胃酸的破坏,制备了碳酸氢钠抗酸泡腾颗粒,并按照制定的新制品原液及成品的质控标准,对原液及成品进行全面检定.结果 制备的口服重组B亚单位O1/O139霍乱疫苗工艺稳定,各项指标均达到质控标准.结论 口服重组B亚单位O1/O139霍乱疫苗安全、有效,且制备工艺可行,符合规模化生产的要求.

  8. Establishment of a method for rapid detection of group O1 and O139 of Vibrio cholera by fluorescence PCR%O1群和O139群霍乱弧菌荧光PCR快速检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    罗可天; 梁晅; 宋妙芳; 丁珊; 林智

    2009-01-01

    Objective To establish a methods for rapid detection of group O1 and O139 of Vibrio cholera by fluorescence PCR. Methods DNA probe and primers were designed based on O antigen code gene of Vibrio.cholera O1 and O139,and methods of simultaneous detection of group O1 and O139 of Vibrio cholera by luorescent PCR was established and the primer,Mg2+ ,dNTP and Taq polyroerase of the system were optimized. The specificity,sensitivity and reproducibility were assessed. Results A fluorescence PCR technique was established for simultaneous detection of group O1 and O139 of Vibrio cholera and the sensitivity of the technique was higher apparently than normal isolation and culture method. Conclusion The fluorescentce PCR technique targatod at detection of O antigen code gene of group O1 and O139 of Vibrio cholera has been established that allowed rapid screening of suspected cholera samples before conventional isolation..%目的 建立检测O1群和0139群霍乱孤菌的实时荧光聚合酶链反应(荧光PCR)方法,并进行优化和评价.方法 根据O1群和O139群霍乱弧菌O抗原编码基因设计探针和引物,建立同时检测霍乱弧菌O1群和O139群的荧光PER方法,并对体系中的引物、Mg2+、dNTP和Taq酶进行优化,然后对建立的方法进行特异性、灵敏性、重复性的评价,并进行224份河口水样本的检测.结果 建立了检测O1群和O139群霍乱孤菌的双重荧光PCR方法,对非O1群和O139群霍乱弧菌无扩增反应,敏感度比常规分离培养高.结论 以O抗原编码基因为目标检测片段建立了O1群和O139群霍乱弧菌双重荧光PCR检测方法,可用于疑似霍乱弧菌感染的样本常规分离前的快速筛查.

  9. Molecular analysis of Pasteurella multocida strains isolated from fowl cholera infection in backyard chickens

    Directory of Open Access Journals (Sweden)

    Mohamed-Wael Abdelazeem Mohamed

    2014-01-01

    Conclusion: Based on the previous findings, there are three spreading clusters that may indicate the association of a small number of P. multocida variants with the majority of cases suggesting that certain clones of P. multocida are able to colonize the examined backyard chickens. Also, the ease and rapidity of RAPD-PCR support the use of this technique as alternative to the more labour-intensive SDS-PAGE system for strain differentiation and epidemiological studies of avian P. multocida. Further application of RAPD technology to the examination of avian cholera outbreaks in commercially available flocks may facilitate more effective management of this disease by providing the potential to investigate correlations of P. multocida genotypes, to identify affiliations between bird types and bacterial genotypes, and to elucidate the role of specific bird species in disease transmission.

  10. Molecular analysis of Pasteurella multocida strains isolated from fowl cholera infection in backyard chickens

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    Objective: To characterize Pasteurella isolated from backyard chickens using whole cell protein lysate profiles and random amplified polymorphic DNA (RAPD) techniques to show their genetic relationship because Pasteurella multocida (P. multocida) is an important cause of fatal infections in backyard chickens. Methods:Twenty one P. multocida isolates were recovered previously from clinical cases of fowl cholera belonging to individual owners and phenotypically analyzed using biochemical tests and serotyping were used for the genetic characterization. Results:Phylogenetic study based on both methods revealed that the recovered population of P. multocida isolated from backyard chickens differs markedly, constituting a well-separated cluster and appearance of 3 distinguishing lineages with greater discrimination shown by RAPD-PCR that resulted in two suclusters in cluster A and three subclusters in cluster B and were related greatly with capsular serogroups for the examined strains. The whole cell protein revealed the presence of dominant protein bands at approximately 41 and 61 kDa in all of the examined isolates that may be a virulent proteins share in the increasing of its pathogenicity. Clear distinctive bands ranged from 123 to 1 554 bp. Conclusions: Based on the previous findings, there are three spreading clusters that may indicate the association of a small number of P. multocida variants with the majority of cases suggesting that certain clones of P. multocida are able to colonize the examined backyard chickens. Also, the ease and rapidity of RAPD-PCR support the use of this technique as alternative to the more labour-intensive SDS-PAGE system for strain differentiation and epidemiological studies of avian P. multocida. Further application of RAPD technology to the examination of avian cholera outbreaks in commercially available flocks may facilitate more effective management of this disease by providing the potential to investigate correlations of P

  11. Transcript changes in Vibrio cholerae in response to salt stress.

    Science.gov (United States)

    Fu, Xiuping; Liang, Weili; Du, Pengcheng; Yan, Meiying; Kan, Biao

    2014-01-01

    Vibrio cholerae, which is a serious human intestinal pathogen, often resides and thrives in estuaries but requires major self-regulation to overcome intestinal hyperosmotic stress or high salt stress in water and food. In the present study, we selected multiple O1 and O139 group V. cholerae strains that were isolated from different regions and during different years to study their salt tolerance. Based on the mechanisms that other bacteria use to respond to high salt stress, we selected salt stress-response related genes to study the mechanisms which V. cholerae responds to high salt stress. V. cholerae strains showed salt-resistance characteristics that varied in salt concentrations from 4% to 6%. However, group O1 and group O139 showed no significant difference in the degree of salt tolerance. The primary responses of bacteria to salt stress, including Na(+) exclusion, K(+) uptake and glutamate biosynthesis, were observed in V. cholerae strains. In addition, some sigma factors were up-regulated in V. cholerae strains, suggesting that V. cholerae may recruit common sigma factors to achieve an active salt stress response. However, some changes in gene transcript levels in response to salt stress in V. cholerae were strain-specific. In particular, hierarchical clustering of differentially expressed genes indicated that transcript levels of these genes were correlated with the degree of salt tolerance. Therefore, elevated transcript levels of some genes, including sigma factors and genes involved in peptidoglycan biosynthesis, may be due to the salt tolerance of strains. In addition, high salt-tolerant strains may recruit common as well as additional sigma factors to activate the salt stress response.

  12. Changing patterns of Vibrio cholerae in sevagram between 1990 and 2005

    Directory of Open Access Journals (Sweden)

    Narang P

    2008-01-01

    Full Text Available Purpose: A retrospective analysis was done to note changes in prevalence, distribution of biotypes, serotypes, antibiotic susceptibility patterns and phage types of Vibrio cholerae isolated in Mahatma Gandhi Institute of Medical Sciences, Sevagram over a period of 16 years. Methods: A total of 535 strains of V. cholerae were isolated from 10,406 stool samples and rectal swabs from January 1990 to December 2005. These comprised of serogroups O1 - 427 (79.89%, O139 - 86 (16.07% and non O1, non O139 - 22 (4.11%. No classical V. cholerae was isolated. Results: Vibrio cholerae serogroup O1 serotype Ogawa was the predominant isolate till 1992. During 1993, serogroup O139 became the main isolate; however, it completely disappeared during 1995-1996 only to reappear in 1997. Serotype Inaba in our area was conspicuous by its absence with only two strains being isolated till June 1999, but during July-December 1999, 11 out of 15 V. cholerae O1 isolates were El Tor Inaba. T4 was the predominant phage type till 1990, T2 during 1991-1994 and T27 (as per the new scheme thereafter. Resistance to tetracycline varied between 2 and 17% for V. cholerae O1. Conclusions: The paper reports on the changing epidemiological markers of V. cholerae isolated from a rural hospital over a period of 16 years.

  13. Microcrustáceos y Vibrio cholerae O1 viable no cultivable (VNC: resultados en la Cuenca del Río Salí, Tucumán, Argentina Microcrustaceans and viable but nonculturable (VNC Vibrio cholerae O1: results in the Salí River basin, Tucumán, Argentina

    Directory of Open Access Journals (Sweden)

    Cecilia Locascio de Mitrovich

    2010-01-01

    Full Text Available Vibrio cholerae reside habitualmente en aguas marinas y continentales. Según las condiciones ambientales y los recursos le sean “favorables” o “desfavorables”, se generan estados viables cultivables (VC o viables no cultivables (VNC respectivamente y, bajo esta última forma sobrevive. Para abordar la problemática del cólera en la Cuenca del Río Salí (Tucumán, Argentina, se realizaron muestreos durante los años 2003-2005 donde se consideraron aspectos fisicos, químicos, biológicos y sanitarios. Para evaluar los probables reservorios del patógeno, se analizó el zooplancton del Río Salí (Canal Norte y Banda Río Salí y Río Lules. La mayor representatividad taxonómica la registraron los copépodos, especialmente Eucyclops neumani (Pesta, 1927, junto a Acanthocyclops robustus (Sars, 1863, Metacyclops sp., Paracyclops chiltoni y Notodiaptomus incompositus (Brian, 1925, además de algunos rotíferos y cladóceros como (Lecane sp., y (Brachionus sp., Moina sp. y Leydigia sp.. La frecuencia de ocurrencia fue baja y no superó el 25%. El Canal Norte fue ambiente más propicio por la riqueza específica, abundancia y constancia de la comunidad. Las variables fisicas y químicas asociadas al zooplancton coincidirían con los valores que por nuestros registros y los antecedentes, se conocen para el desarrollo del patógeno. En el período estival hubo coincidencia entre la presencia de la forma VNC de V. cholerae O1 (inmunofluorescencia con anticuerpos anti O1 y el desarrollo del zooplancton. Se observaron formas VNC sobre apéndices o estructuras de copépodos ciclopoideos y cladóceros quidóridos, reflejando probablemente afinidad con sustratos quitinosos.Vibrio cholerae habitually lives in marine and continental waters. According to "favourable" or "unfavourable" resources and environmental conditions, viable (VC or viable non-culturable (VNC states will be generated, surviving only the latter form. To address the problem of

  14. CHANGING EPIDEMIOLOGICAL TREND OF CHOLERA IN WEST BENGAL: THE GIANT IS BACK

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    Indrani

    2013-11-01

    Full Text Available ABSTRACT: Choler a is a devastating diarrheal disease caused by V. cholera. Two biotypes of V. cholerae O1, classical and El - Tor, are distinguished. Each biotype is further subdivided into two serotypes, termed Inaba and Ogawa. As large deltaic areas of the Ganges and Brah maputra rivers are considered to be the homeland of cholera, objective of our study was to detect the circulating strain of Vibrio causing Cholera outbreaks in different pockets of West Bengal. Water samples collected from the water sources of the affected areas and stool samples and or rectal swabs of suspected cases were tested according to standard bacteriological protocol. In July 2008, Cholera outbreak was caused by Vibrio cholera e O1 El - Tor Inaba serotype in Darjeeling district affecting 176 persons. Mean age of the affected people was 15years. Thereafter in the June, 2010, there was an outbreak of Vibrio cholerae El Tor Ogawa affecting 87 people in Maldah. Mean age of the cases was 25years. Similar type of Vibrio cholerae O1 El - Tor Ogawa strain was is olated in the outbreak of June to September 2012 in Maldah with lower mean age of cases i.e.7years. A total of 93 patients suffered from Cholera during this outbreak. Cholera outbreak in West Bengal was caused by classical Vibrio cholerae O1 Ogawa serotype affecting 803 people in North 24 Paragana in October, 2013. Mean age of the patients was 31year. As classical Vibrio causes more severe disease than El - Tor, its reemergence with multidrug resistant properties is no doubt an upcoming threat

  15. Changing genotypes of cholera toxin (CT) of Vibrio cholerae O139 in Bangladesh and description of three new CT genotypes.

    Science.gov (United States)

    Bhuiyan, Nurul A; Nusrin, Suraia; Alam, Munirul; Morita, Masatomo; Watanabe, Haruo; Ramamurthy, Thandavarayan; Cravioto, Alejandro; Nair, Gopinath Balakrish

    2009-11-01

    We determined the genotype of cholera toxin by amplifying and sequencing the B-subunit in a sequential collection of 90 strains of Vibrio cholerae O139 isolated over the past 13 years since its first description in 1992. Representative strains isolated during 1993-1997 harboured ctxB of El Tor type (genotype 3). Twenty-six strains isolated during 1999, 2001, 2005 and three strains isolated in 1998, 2000 and 2002 were identified to belong to new ctxB genotypes 4 and 5, respectively. Genotype 5 was similar to genotype 1 except at position 28 (D-->A). The genotype 6 was similar to genotype 4 except at position 34 (H-->P). The implication of switch in terms of function of the toxin and its impact on human disease is unclear. How this change has influenced their prevalence relative to that of V. cholerae O1 in human infection is also not clear. The other common virulence gene clusters including the Vibrio pathogenicity island-1, Vibrio seventh pandemic island (VSP)-I and VSP-II of V. cholerae O139 did not show any remarkable difference from that of the O1 El Tor strains. Overall, the majority of the O139 strains tested in this study were similar to the El Tor strains but had altered ctxB genotype. This change and the impact that it causes to the epidemiology of cholera caused by O139 should be closely monitored.

  16. In Vitro Inhibition of Cholera Toxin Production in Vibrio cholerae by Methanol Extract of Sweet Fennel Seeds and Its Components.

    Science.gov (United States)

    Chatterjee, Shruti; Zahid, M Shamim Hasan; Awasthi, Sharda Prasad; Chowdhury, Nityananda; Asakura, Masahiro; Hinenoya, Atsushi; Ramamurthy, T; Iwaoka, Emiko; Aoki, Shunji; Yamasaki, Shinji

    2016-09-21

    A newly emerged Vibrio cholerae O1 El Tor variant strain with multidrug resistance is considered a threat to public health. Recent strategies to suppress virulence factors production instead of bacterial growth may lead to less selective pressure for the emergence of resistant strains. The use of spices and their active constituents as the inhibitory agents against cholera toxin (CT) production in V. cholerae may be an alternative approach to treat cholera. In this study, we examined the potential of sweet fennel seed (Foeniculum vulgare Miller var. dulce) methanol extract to inhibit CT production in V. cholerae without affecting viability. The methanol extract of sweet fennel seeds significantly inhibited CT production in various V. cholerae strains, regardless of serogroup or biotype. Interestingly, trans-anethole and 4-allylanisole, essential oil components of sweet fennel seeds, also demonstrated similar effects. Here, we report that sub-bactericidal concentrations of sweet fennel seed methanol extract and its major components can drastically inhibit CT production in various V. cholerae strains.

  17. Characterization of phenotype and molecular characteristics on Vibrio cholerae strains isolated in Shanghai, 1962-2011%上海市1962-2011年霍乱弧菌表型特征及分子分型研究

    Institute of Scientific and Technical Information of China (English)

    屠丽红; 陈敏; 李伟; 张曦; 陈洪友; 王文静

    2013-01-01

    Objective To describe the phenotype and molecular characteristics of Vibrio (V.) cholerae strains isolated in Shanghai,from 1962 to 2011.Methods K-B test was used to investigate the antibiotic resistance of V.cholerae strains.PCR was applied to detect seven virulence-related genes including cholera toxin (ctxA),zonula oecludens toxin (zot),accessory cholera enterotoxin (ace),hemolysin (hlyA),toxin-coregulated pilus (tcpA) outer membrane protein (ompU) and the regulatory protein genes (toxR).Genetic relation was assessed by pulsed-field gel electrophoresis (PFGE) and the patterns were clustered by BioNumerics software.Results V.cholerae strains isolated from 1962 to 1996 were sensitive to most of the antibiotics.However,the strains isolated from 2005 to 2011 were resistant to many antibiotics.V.cholerae O 139 group showed higher prevalence of resistance to several antibiotics compared with O l group,and the resistance rate of the O139 toxigenic isolates was higher than that of the non-toxigenic isolates.Most of the O1 strains isolated from 2005 to 2011 were non-toxigenic while O139 strains isolated from 2005 to 2011 were almost toxigenic.There were no strains ofctxA+ detected from the rivers from 2005 to 2011.Main gene type of the O1 strains detected from the aquatic products was hlyA+toxR+ompU+,while that of the O139 strains was hlyA+toxR+ompU+ ctxA + ace +zot + tcpA +.Using PFGE,222 V.cholerae strains were subtyped into 121 molecular types.O139 strains were divided to three clusters and O1 strains to five clusters.Conclusion The characteristics of V.cholerae strains isolated in Shanghai from 1962 to 2011 showed great changes,suggesting that more attention should be paid to the multiplication on antibiotic resistance of V.cholerae strains.%目的 分析1962-2011年上海市霍乱弧菌的表型及分子分型特征.方法 采用WHO推荐的改良K-B纸片法,对222株霍乱弧菌进行11种抗菌药物(头孢曲松、强力霉素、诺氟沙星、环丙沙星、

  18. Survivability and molecular variation in Vibrio cholerae from epidemic sites in China.

    Science.gov (United States)

    Li, X Q; Wang, M; Deng, Z A; Shen, J C; Zhang, X Q; Liu, Y F; Cai, Y S; Wu, X W; DI, B

    2015-01-01

    The survival behaviour of Vibrio cholerae in cholera epidemics, together with its attributes of virulence-associated genes and molecular fingerprints, are significant for managing cholera epidemics. Here, we selected five strains representative of V. cholerae O1 and O139 involved in cholera events, examined their survival capacity in large volumes of water sampled from epidemic sites of a 2005 cholera outbreak, and determined virulence-associated genes and molecular subtype changes of the surviving isolates recovered. The five strains exhibited different survival capacities varying from 17 to 38 days. The virulence-associated genes of the surviving isolates remained unchanged, while their pulsotypes underwent slight variation. In particular, one waterway-isolated strain maintained virulence-associated genes and evolved to share the same pulsotype as patient strains, highlighting its role in the cholera outbreak. The strong survival capacity and molecular attributes of V. cholerae might account for its persistence in environmental waters and the long duration of the cholera outbreak, allowing effective control measures.

  19. Detection of viable and viable nonculturable Vibrio cholerae O1 through cultures and immunofluorescence in the Tucumán rivers, Argentina Detecção de Vibrio cholerae O1 viável e viável não cultivável, através de técnicas de cultivo e imunofluorescência nos rios de Tucumán, Argentina

    Directory of Open Access Journals (Sweden)

    Olga Aulet

    2007-08-01

    Full Text Available Vibrio cholerae has been sporadically isolated from rivers in Tucumán, Argentina, since the outbreak in 1991. The aim of this study was to determine the environmental reservoir of the bacterium in these rivers, assessing the presence of Vibrio cholerae non-O1 and O1 (the latter both in its viable culturable and non culturable state and its relationship to environmental physicochemical variables. 18 water samplings were collected in the Salí River (in Canal Norte and Banda and the Lules River between 2003 and 2005. Physical-chemical measurements (pH, water temperature, electrical conductivity and dissolved oxygen were examined. Vibrio cholerae was investigated with conventional culture methods and with Direct Immunofluorescence (DFA-VNC in order to detect viable non culturable organisms. All isolated microorganisms corresponded to Vibrio cholerae non-O1 and non-O139 (Lules 26%, Canal Norte 33% and Banda 41%. The majority was found during spring and summer and correlated with temperature and pH. Non culturable Vibrio cholerae O1 was detected year round in 38 of the 54 water samples analyzed. Application of the Pearson correlation coefficient revealed that there was no relationship between positive immunofluorescence results and environmental physicochemical parameters. Genes coding for somatic antigen O1 were confirmed in all DFA-VNC-positive samples, whereas the virulence-associated ctxA and tcpA genes were confirmed in 24 samples.Vibrio cholerae tem sido isolado esporadicamente nos rios da Província de Tucumán, Argentina, desde outubro de 1991. O objetivo deste estudo foi localizar os reservatórios nestes rios, identificar a presença de Vibrio cholerae O1 (em estado cultivável e não cultivável e relacionar a presença desta bactéria com as variações físico-químicos da água. Foram coletadas dezoito amostras de água do rio Salí (nas localidades de Canal Norte e Banda e do rio Lules, entre 2003 e 2005. Estas foram submetidas a an

  20. Vibrio cholerae in an Historically Cholera-Free Country.

    Science.gov (United States)

    Haley, Bradd J; Chen, Arlene; Grim, Christopher J; Clark, Philip; Diaz, Celia Municio; Taviani, Elisa; Hasan, Nur A; Sancomb, Elizabeth; Elnemr, Wessam Mahmoud; Islam, Muhammad A; Huq, Anwar; Colwell, Rita R; Benediktsdóttir, Eva

    2012-08-01

    We report the autochthonous existence of Vibrio cholerae in coastal waters of Iceland, a geothermally active country where cholera is absent and has never been reported. Seawater, mussel, and macroalgae samples were collected close to and distant from sites where geothermal activity causes a significant increase in water temperature during low tides. V. cholerae was detected only at geothermal-influenced sites during low-tides. None of the V. cholerae isolates encoded cholera toxin (ctxAB) and all were non-O1/non-O139 serogroups. However, all isolates encoded other virulence factors that are associated with cholera as well as extra-intestinal V. cholerae infections. The virulence factors were functional at temperatures of coastal waters of Iceland, suggesting an ecological role. It is noteworthy that V. cholerae was isolated from samples collected at sites distant from anthropogenic influence, supporting the conclusion that V. cholerae is autochthonous to the aquatic environment of Iceland.

  1. The antigenic relationship between Brettanomyces-Debaryomyces strains and the Salmonella cholerae-suis O antigen.

    Science.gov (United States)

    Aksoycan, N; Sağanak, I; Wells, G

    1978-01-01

    The immune sera for Brettanomyces lambicus, B. claussenii, Debaryomyces hansenii and D. marama agglutinated Salmonella cholerae-suis (0:6(2), 7). The immune serum for S. cholerae-suis agglutinated B. lambicus, B. clausenni, D. hansenii and D. marama. Absorption and agglutination cross-tested demonstrated common antigen factor(s) in the tested yeasts and Salmonella 0:7 antigen.

  2. Prevalence and characterization of Vibrio cholerae isolated from shrimp products imported into Denmark

    DEFF Research Database (Denmark)

    Dalsgaard, A.; Bjergskov, T.; Jeppesen, V.F.

    1996-01-01

    A total of 3,555 metric tonnes of warm water shrimp were imported into Denmark from December 1994 to July 1995. V. cholerae O1 was not detected in any of the 748 samples analyzed. Non-Ol V. cholerae was found in a single (0.1%) cooked frozen shrimp product and in five (0.7%) raw frozen products...... contained plasmids or genes encoding cholera toxin (CT) or heat-stable enterotoxin (NAG-ST), The absence of V. cholerae O1 and the low number of samples containing CT and NAG-ST negative non-Ol strains in imported shrimp suggest that I! cholerae in such products may not constitute a public health problem....

  3. Swedish isolates of Vibrio cholerae enhance their survival when interacted intracellularly with Acanthamoeba castellanii

    OpenAIRE

    Shanan, Salah; Bayoumi, Magdi; Saeed, Amir; Sandström, Gunnar; Abd, Hadi

    2016-01-01

    Vibrio cholerae is a Gram-negative bacterium that occurs naturally in aquatic environment. Only V. cholerae O1 and V. cholerae O139 produce cholera toxin and cause cholera, other serogroups can cause gastroenteritis, open wounds infection, and septicaemia. V. cholerae O1 and V. cholerae O139 grow and survive inside Acanthamoeba castellanii. The aim of this study is to investigate the interactions of the Swedish clinical isolates V. cholerae O3, V. cholerae O4, V. cholerae O5, V. cholerae O11,...

  4. Cholera outbreaks in South and Southeast Asia: descriptive analysis, 2003-2012.

    Science.gov (United States)

    Mahapatra, Tanmay; Mahapatra, Sanchita; Babu, Giridhara R; Tang, Weiming; Banerjee, Barnali; Mahapatra, Umakanta; Das, Aritra

    2014-01-01

    We conducted descriptive analysis of available information regarding the epidemiology of cholera outbreaks in South and Southeast Asia during 2003-2012. Information from 58 articles, 8 reports, and World Health Organization databases were analyzed. Overall, 113 cholera outbreaks were studied in South and Southeast Asia during the past 10 years. The majority of the outbreaks (69%) occurred in Southeast Asia, including India (52%). The highest number of outbreaks was observed in 2004 (25.7%). The most commonly identified source was contaminated water: however, in some countries, the spread of cholera was facilitated via contaminated seafood (e.g., Myanmar, Thailand, and Singapore). Several genotypes and phenotypes of Vibrio cholerae, the causative agent of cholera, were identified in the outbreaks, including V. cholerae O1 El Tor (Ogawa and Inaba) and V. cholerae O139. The emergence of multidrug-resistant V. cholerae strains was a major concern. Cholera-related mortality was found to be low across the outbreaks, except in Orissa, India (currently Odisha) during 2007, where the case fatality rate was 8.6%. Potential limitations included underreporting, discrepancies, possible exclusion of nonindexed reports, and incomprehensive search terms. The provision of safe water and proper sanitation appear to be critical for the control of further spread of cholera in South Asian and Southeast Asian regions.

  5. Seasonal dynamics of Vibrio cholerae and its phages in riverine ecosystem of Gangetic West Bengal: cholera paradigm.

    Science.gov (United States)

    Mookerjee, Subham; Jaiswal, Abhishek; Batabyal, Prasenjit; Einsporn, Marc H; Lara, Ruben J; Sarkar, Banwarilal; Neogi, Sucharit Basu; Palit, Anup

    2014-10-01

    The Gangetic delta is a century-old cholera endemic belt where the role of riverine-estuarine ecosystem in cholera transmission has never been elucidated. Seasonality, distribution, and abundance of environmental Vibrio cholerae O1/O139 and vibriophage in Hooghly riverine-estuarine environment and their correlation with cholera incidence pattern in West Bengal, India, have been analyzed for the first time across summer, monsoon, and winter months. A total of 146 water samples collected from two sites of the Hooghly River (Howrah and Diamond Harbour) were analyzed physicochemically along with cultivable Vibrio count (CVC), V. cholerae O1/O139, and vibriophages. V. cholerae O1 was detected in 56 (38.3%) samples, while 66 (45.2%) were positive for V. cholerae O1 phages. Flood tide, water temperature (31 ± 1.6 °C), and turbidity (≥250 nephelometric turbidity unit (NTU)) significantly stimulated V. cholerae and vibriophage abundance in riverine ecosystem. Solitary existence of V. cholerae O1 and phages (p V. cholerae O1 or V. cholerae O1 Φ) on the other. Significant association (p cholera cases and V. cholerae O1 in aquatic environment implies the role of riverine-estuarine ecosystem in cholera transmission. A "biomonitoring tool" of physicochemical stimulants, tidal, and climatic variants has been proposed collating V. cholerae and phage dynamics that can forewarn any impending cholera outbreak.

  6. Occurrence in Mexico, 1998-2008, of Vibrio cholerae CTX+ El Tor carrying an additional truncated CTX prophage.

    Science.gov (United States)

    Alam, Munirul; Rashed, Shah Manzur; Mannan, Shahnewaj Bin; Islam, Tarequl; Lizarraga-Partida, Marcial Leonardo; Delgado, Gabriela; Morales-Espinosa, Rosario; Mendez, Jose Luis; Navarro, Armando; Watanabe, Haruo; Ohnishi, Makoto; Hasan, Nur A; Huq, Anwar; Sack, R Bradley; Colwell, Rita R; Cravioto, Alejandro

    2014-07-08

    The seventh cholera pandemic caused by Vibrio cholerae O1 El Tor (ET) has been superseded in Asia and Africa by altered ET possessing the cholera toxin (CTX) gene of classical (CL) biotype. The CL biotype of V. cholerae was isolated, along with prototypic and altered ET, during the 1991 cholera epidemic in Mexico and subsequently remained endemic until 1997. Microbiological, molecular, and phylogenetic analyses of clinical and environmental V. cholerae isolated in Mexico between 1998 and 2008 revealed important genetic events favoring predominance of ET over CL and altered ET. V. cholerae altered ET was predominant after 1991 but not after 2000. V. cholerae strains isolated between 2001 and 2003 and a majority isolated in 2004 lacked CTX prophage (Φ) genes encoding CTX subunits A and B and repeat sequence transcriptional regulators of ET and CL biotypes: i.e., CTXΦ(-). Most CTXΦ(-) V. cholerae isolated in Mexico between 2001 and 2003 also lacked toxin coregulated pili tcpA whereas some carried either tcpA(ET) or a variant tcpA with noticeable sequence dissimilarity from tcpA(CL). The tcpA variants were not detected in 2005 after CTXΦ(+) ET became dominant. All clinical and environmental V. cholerae O1 strains isolated during 2005-2008 in Mexico were CTXΦ(+) ET, carrying an additional truncated CTXΦ instead of RS1 satellite phage. Despite V. cholerae CTXΦ(-) ET exhibiting heterogeneity in pulsed-field gel electrophoresis patterns, CTXΦ(+) ET isolated during 2004-2008 displayed homogeneity and clonal relationship with V. cholerae ET N16961 and V. cholerae ET isolated in Peru.

  7. Study about the sensibility in vitro of different strains of Vibrio cholera 01 exposed to 60 Co gamma radiation; Estudo da sensibilidade in vitro de diferentes cepas de Vibio cholerae 01 a radiacao gama de 60Co

    Energy Technology Data Exchange (ETDEWEB)

    Moraes, Ivany Rodrigues de [Instituto Adolfo Lutz, Sao Paulo, SP (Brazil). Servicos de Saude; Gelli, Dilma Scala; Jakabi, Miyoko [Instituto Adolfo Lutz, Sao Paulo, SP (Brazil). Secao de Microbiologia; Mastro, Nelida Lucia del [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)]. E-mail: nlmastro@net.ipen.br

    1998-07-01

    The presence of some microorganisms in food, or the metabolites originated during their own multiplication may bring several diseases to humans: intoxications and food borne infections. Among the agents that may cause those diseases, we find Vibrio cholerae 01. In this experiment, the studies are focused on the radiosensibility in vitro of four strains of V. cholerae 01, exposed to different doses of ionizing radiation of {sup 60} Co. The results are compared with other data related to bacterial food borne diseases, including water. (author)

  8. Cholera Toxin Production Induced upon Anaerobic Respiration is Suppressed by Glucose Fermentation in Vibrio cholerae.

    Science.gov (United States)

    Oh, Young Taek; Lee, Kang-Mu; Bari, Wasimul; Kim, Hwa Young; Kim, Hye Jin; Yoon, Sang Sun

    2016-03-01

    The causative agent of pandemic cholera, Vibrio cholerae, infects the anaerobic environment of the human intestine. Production of cholera toxin (CT), a major virulence factor of V. cholerae, is highly induced during anaerobic respiration with trimethylamine N-oxide (TMAO) as an alternative electron acceptor. However, the molecular mechanism of TMAO-stimulated CT production is not fully understood. Herein, we reveal that CT production during anaerobic TMAO respiration is affected by glucose fermentation. When the seventh pandemic V. cholerae O1 strain N16961 was grown with TMAO and additional glucose, CT production was markedly reduced. Furthermore, an N16961 Δcrp mutant, devoid of cyclic AMP receptor protein (CRP), was defective in CT production during growth by anaerobic TMAO respiration, further suggesting a role of glucose metabolism in regulating TMAO-mediated CT production. TMAO reductase activity was noticeably decreased when grown together with glucose or by mutation of the crp gene. A CRP binding region was identified in the promoter region of the torD gene, which encodes a structural subunit of the TMAO reductase. Gel shift assays further confirmed the binding of purified CRP to the torD promoter sequence. Together, our results suggest that the bacterial ability to respire using TMAO is controlled by CRP, whose activity is dependent on glucose availability. Our results reveal a novel mechanism for the regulation of major virulence factor production by V. cholerae under anaerobic growth conditions.

  9. Seroepidemiologic survey of epidemic cholera in Haiti to assess spectrum of illness and risk factors for severe disease.

    Science.gov (United States)

    Jackson, Brendan R; Talkington, Deborah F; Pruckler, James M; Fouché, M D Bernadette; Lafosse, Elsie; Nygren, Benjamin; Gómez, Gerardo A; Dahourou, Georges A; Archer, W Roodly; Payne, Amanda B; Hooper, W Craig; Tappero, Jordan W; Derado, Gordana; Magloire, Roc; Gerner-Smidt, Peter; Freeman, Nicole; Boncy, Jacques; Mintz, Eric D

    2013-10-01

    To assess the spectrum of illness from toxigenic Vibrio cholerae O1 and risk factors for severe cholera in Haiti, we conducted a cross-sectional survey in a rural commune with more than 21,000 residents. During March 22-April 6, 2011, we interviewed 2,622 residents ≥ 2 years of age and tested serum specimens from 2,527 (96%) participants for vibriocidal and antibodies against cholera toxin; 18% of participants reported a cholera diagnosis, 39% had vibriocidal titers ≥ 320, and 64% had vibriocidal titers ≥ 80, suggesting widespread infection. Among seropositive participants (vibriocidal titers ≥ 320), 74.5% reported no diarrhea and 9.0% had severe cholera (reported receiving intravenous fluids and overnight hospitalization). This high burden of severe cholera is likely explained by the lack of pre-existing immunity in this population, although the virulence of the atypical El Tor strain causing the epidemic and other factors might also play a role.

  10. Molecular Epidemiology and Antibiotic Susceptibility of Vibrio cholerae Associated with a Large Cholera Outbreak in Ghana in 2014.

    Directory of Open Access Journals (Sweden)

    Daniel Eibach

    2016-05-01

    Full Text Available Ghana is affected by regular cholera epidemics and an annual average of 3,066 cases since 2000. In 2014, Ghana experienced one of its largest cholera outbreaks within a decade with more than 20,000 notified infections. In order to attribute this rise in cases to a newly emerging strain or to multiple simultaneous outbreaks involving multi-clonal strains, outbreak isolates were characterized, subtyped and compared to previous epidemics in 2011 and 2012.Serotypes, biotypes, antibiotic susceptibilities were determined for 92 Vibrio cholerae isolates collected in 2011, 2012 and 2014 from Southern Ghana. For a subgroup of 45 isolates pulsed-field gel electrophoresis, multilocus sequence typing and multilocus-variable tandem repeat analysis (MLVA were performed. Eighty-nine isolates (97% were identified as ctxB (classical type positive V. cholerae O1 biotype El Tor and three (3% isolates were cholera toxin negative non-O1/non-O139 V. cholerae. Among the selected isolates only sulfamethoxazole/trimethoprim resistance was detectable in 2011, while 95% of all 2014 isolates showed resistance towards sulfamethoxazole/trimethoprim, ampicillin and reduced susceptibility to ciprofloxacin. MLVA achieved the highest subtype discrimination, revealing 22 genotypes with one major outbreak cluster in each of the three outbreak years. Apart from those clusters genetically distant genotypes circulate during each annual epidemic.This analysis suggests different endemic reservoirs of V. cholerae in Ghana with distinct annual outbreak clusters accompanied by the occurrence of genetically distant genotypes. Preventive measures for cholera transmission should focus on aquatic reservoirs. Rapidly emerging multidrug resistance must be monitored closely.

  11. Reporte histórico: Primer Aislamiento de Vibrio cholera serogrupo O1 biovar El Tor serovar Inaba durante la epidemia de cólera en el Perú ‑ 1991 Historical report: first isolation of Vibrio cholera serogroup O1 biovar El Tor serovar Inaba during the cholerae epidemic in Perú ‑ 1991

    OpenAIRE

    Nora Bravo Cruz; Alfredo Guillén

    2011-01-01

    Hace 20 años apareció una enfermedad diarreica nueva en el Perú y el Laboratorio de Referencia de Enteropatógenos del Instituto Nacional de Salud, cumplió una labor destacada en el aislamiento e identificación rápida y oportuna del Vibrio cholerae. La enfermedad del cólera no se había presentado anteriormente, pero en la última semana de enero de 1991 se detectó un brote epidémico de diarrea aguda con deshidratación intensa y algunos casos de fallecidos. La epidemia afectó, al comienzo, varia...

  12. Tagging a Vibrio cholerae El Tor candidate vaccine strain by disruption of its hemagglutinin/protease gene using a novel reporter enzyme: Clostridium thermocellum endoglucanase A.

    Science.gov (United States)

    Robert, A; Silva, A; Benitez, J A; Rodriguez, B L; Fando, R; Campos, J; Sengupta, D K; Boesman-Finkelstein, M; Finkelstein, R A

    1996-11-01

    The celA gene encoding Clostridium thermocellum endoglucanase A was expressed in Vibrio cholerae on its own promoter and used to tag a candidate El Tor biotype cholera vaccine strain. Colonies of the tagged strain could be unequivocally distinguished by overlaying them with CM-cellulose indicator agar and Congo Red staining. Expression of celA did not affect growth of V. cholerae in vitro and in vivo. The celA gene was inserted in the chromosomal hap locus encoding V. cholerae hemagglutinin/protease, a putative "detachase", to create a hap- mutant that could be identified and scored by its halo of cellulolytic activity. The inactivation of hap had a positive effect on colonization in the infant mice model. The above results indicate that celA is a suitable marker gene for V. cholerae and hap is an appropriate locus for insertion of foreign DNA in vaccine development. Inactivation of hap, by increasing the duration of adherence, might decrease excretion of the resulting vaccine vector strain and thus increase its immunogenicity.

  13. Vibrio cholerae laboratory infection of the adult house fly Musca domestica.

    Science.gov (United States)

    El-Bassiony, G M; Luizzi, V; Nguyen, D; Stoffolano, J G; Purdy, A E

    2016-12-01

    The present study was designed to test the hypothesis that house flies may be capable of specifically harbouring ingested Vibrio cholerae in their digestive tracts. Flies were continuously fed green fluorescent protein (GFP)-labelled, non-O1/non-O139 environmental strains of V. cholerae. Bacterial burdens were quantitatively measured using plate counts and localization was directly observed using confocal microscopy. Vibrio cholerae were present in the fly alimentary canal after just 4 h, and reached a plateau of ∼10(7) colony-forming units (CFU)/fly after 5 days in those flies most tolerant of the pathogen. However, individual flies were resistant to the pathogen: one or more flies were found to carry V. cholerae CFU at each time-point examined. In flies carrying V. cholerae, the pathogen was predominantly localized to the midgut rather than the rectal space or crop. The proportion of house flies carrying V. cholerae in the midgut was dose-dependent: the continuous ingestion of a concentrated, freshly prepared dose of V. cholerae increased the likelihood that fluorescent cells would be observed. However, V. cholerae may be a transient inhabitant of the house fly. This work represents the first demonstration that V. cholerae can inhabit the house fly midgut, and provides a platform for future studies of host, pathogen and environmental mediators of the successful colonization of this disease vector.

  14. The Vaccine Candidate Vibrio cholerae 638 Is Protective against Cholera in Healthy Volunteers

    Science.gov (United States)

    García, Luis; Jidy, Manuel Díaz; García, Hilda; Rodríguez, Boris L.; Fernández, Roberto; Año, Gemma; Cedré, Bárbara; Valmaseda, Tania; Suzarte, Edith; Ramírez, Margarita; Pino, Yadira; Campos, Javier; Menéndez, Jorge; Valera, Rodrigo; González, Daniel; González, Irma; Pérez, Oliver; Serrano, Teresita; Lastre, Miriam; Miralles, Fernando; del Campo, Judith; Maestre, Jorge Luis; Pérez, José Luis; Talavera, Arturo; Pérez, Antonio; Marrero, Karen; Ledón, Talena; Fando, Rafael

    2005-01-01

    Vibrio cholerae 638 is a living candidate cholera vaccine strain attenuated by deletion of the CTXΦ prophage from C7258 (O1, El Tor Ogawa) and by insertion of the Clostridium thermocellum endoglucanase A gene into the hemagglutinin/protease coding sequence. This vaccine candidate was previously found to be well tolerated and immunogenic in volunteers. This article reports a randomized, double-blind, placebo-controlled trial conducted to test short-term protection conferred by 638 against subsequent V. cholerae infection and disease in volunteers in Cuba. A total of 45 subjects were enrolled and assigned to receive vaccine or placebo. The vaccine contained 109 CFU of freshly harvested 638 buffered with 1.3% NaHCO3, while the placebo was buffer alone. After vaccine but not after placebo intake, 96% of volunteers had at least a fourfold increase in vibriocidal antibody titers, and 50% showed a doubling of at least the lipopolysaccharide-specific immunoglobulin A titers in serum. At 1 month after vaccination, five volunteers from the vaccine group and five from the placebo group underwent an exploratory challenge study with 109 CFU of ΔCTXΦ attenuated mutant strain V. cholerae 81. Only two volunteers from the vaccine group shed strain 81 in their feces, but none of them experienced diarrhea; in the placebo group, all volunteers excreted the challenge strain, and three had reactogenic diarrhea. An additional 12 vaccinees and 9 placebo recipients underwent challenge with 7 × 105 CFU of virulent strain V. cholerae 3008 freshly harvested from a brain heart infusion agar plate and buffered with 1.3% NaHCO3. Three volunteers (25%) from the vaccine group and all from the placebo group shed the challenge agent in their feces. None of the 12 vaccinees but 7 volunteers from the placebo group had diarrhea, and 2 of the latter exhibited severe cholera (>5,000 g of diarrheal stool). These results indicate that at 1 month after ingestion of a single oral dose (109 CFU) of strain

  15. The Vibrio cholerae cytolysin promotes chloride secretion from intact human intestinal mucosa.

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    Lucantonio Debellis

    Full Text Available BACKGROUND: The pathogenicity of the Vibrio cholerae strains belonging to serogroup O1 and O139 is due to the production of virulence factors such as cholera toxin (CT and the toxin-coregulated pilus (TCP. The remaining serogroups, which mostly lack CT and TCP, are more frequently isolated from aquatic environmental sources than from clinical samples; nevertheless, these strains have been reported to cause human disease, such as sporadic outbreaks of watery diarrhoea and inflammatory enterocolitis. This evidence suggested the possibility that other virulence factor(s than cholera toxin might be crucial in the pathogenesis of Vibrio cholerae-induced diarrhoea, but their nature remains unknown. VCC, the hemolysin produced by virtually all Vibrio cholerae strains, has been proposed as a possible candidate, though a clear-cut demonstration attesting VCC as crucial in the pathogenesis of Vibrio cholerae-induced diarrhoea is still lacking. METHODOLOGY/PRINCIPAL FINDINGS: Electrophysiological parameters and paracellular permeability of stripped human healthy colon tissues, obtained at subtotal colectomy, mounted in Ussing chamber were studied in the presence or absence of VCC purified from culture supernatants of V. cholerae O1 El Tor strain. Short circuit current (I(SC and transepithelial resistance (R(T were measured by a computerized voltage clamp system. The exposure of sigmoid colon specimens to 1 nM VCC resulted in an increase of I(SC by 20.7%, with respect to the basal values, while R(T was reduced by 12.3%. Moreover, increase in I(SC was abolished by bilateral Cl(- reduction. CONCLUSION/SIGNIFICANCE: Our results demonstrate that VCC, by forming anion channels on the apical membrane of enterocytes, triggers an outward transcellular flux of chloride. Such an ion movement, associated with the outward movement of Na(+ and water, might be responsible for the diarrhoea caused by the non-toxigenic strains of Vibrio cholerae.

  16. Vibrio cholerae Infection of Drosophilamelanogaster Mimics the Human Disease Cholera.

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    2005-09-01

    Full Text Available Cholera, the pandemic diarrheal disease caused by the gram-negative bacterium Vibrio cholerae, continues to be a major public health challenge in the developing world. Cholera toxin, which is responsible for the voluminous stools of cholera, causes constitutive activation of adenylyl cyclase, resulting in the export of ions into the intestinal lumen. Environmental studies have demonstrated a close association between V. cholerae and many species of arthropods including insects. Here we report the susceptibility of the fruit fly, Drosophila melanogaster, to oral V. cholerae infection through a process that exhibits many of the hallmarks of human disease: (i death of the fly is dependent on the presence of cholera toxin and is preceded by rapid weight loss; (ii flies harboring mutant alleles of either adenylyl cyclase, Gsalpha, or the Gardos K channel homolog SK are resistant to V. cholerae infection; and (iii ingestion of a K channel blocker along with V. cholerae protects wild-type flies against death. In mammals, ingestion of as little as 25 mug of cholera toxin results in massive diarrhea. In contrast, we found that ingestion of cholera toxin was not lethal to the fly. However, when cholera toxin was co-administered with a pathogenic strain of V. cholerae carrying a chromosomal deletion of the genes encoding cholera toxin, death of the fly ensued. These findings suggest that additional virulence factors are required for intoxication of the fly that may not be essential for intoxication of mammals. Furthermore, we demonstrate for the first time the mechanism of action of cholera toxin in a whole organism and the utility of D. melanogaster as an accurate, inexpensive model for elucidation of host susceptibility to cholera.

  17. Proteins involved in difference of sorbitol fermentation rates of the toxigenic and nontoxigenic Vibrio cholerae El Tor strains revealed by comparative proteome analysis

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    Kan Biao

    2009-07-01

    Full Text Available Abstract Background The nontoxigenic V. cholerae El Tor strains ferment sorbitol faster than the toxigenic strains, hence fast-fermenting and slow-fermenting strains are defined by sorbitol fermentation test. This test has been used for more than 40 years in cholera surveillance and strain analysis in China. Understanding of the mechanisms of sorbitol metabolism of the toxigenic and nontoxigenic strains may help to explore the genome and metabolism divergence in these strains. Here we used comparative proteomic analysis to find the proteins which may be involved in such metabolic difference. Results We found the production of formate and lactic acid in the sorbitol fermentation medium of the nontoxigenic strain was earlier than of the toxigenic strain. We compared the protein expression profiles of the toxigenic strain N16961 and nontoxigenic strain JS32 cultured in sorbitol fermentation medium, by using fructose fermentation medium as the control. Seventy-three differential protein spots were found and further identified by MALDI-MS. The difference of product of fructose-specific IIA/FPR component gene and mannitol-1-P dehydrogenase, may be involved in the difference of sorbitol transportation and dehydrogenation in the sorbitol fast- and slow-fermenting strains. The difference of the relative transcription levels of pyruvate formate-lyase to pyruvate dehydrogenase between the toxigenic and nontoxigenic strains may be also responsible for the time and ability difference of formate production between these strains. Conclusion Multiple factors involved in different metabolism steps may affect the sorbitol fermentation in the toxigenic and nontoxigenic strains of V. cholerae El Tor.

  18. Multiple antibiotic resistance of Vibrio cholerae serogroup O139 in China from 1993 to 2009.

    Science.gov (United States)

    Yu, Li; Zhou, Yanyan; Wang, Ruibai; Lou, Jing; Zhang, Lijuan; Li, Jie; Bi, Zhenqiang; Kan, Biao

    2012-01-01

    Regarded as an emerging diarrheal micropathogen, Vibrio cholerae serogroup O139 was first identified in 1992 and has become an important cause of cholera epidemics over the last two decades. O139 strains have been continually isolated since O139 cholera appeared in China in 1993, from sporadic cases and dispersed foodborne outbreaks, which are the common epidemic types of O139 cholera in China. Antibiotic resistance profiles of these epidemic strains are required for development of clinical treatments, epidemiological studies and disease control. In this study, a comprehensive investigation of the antibiotic resistance of V. cholerae O139 strains isolated in China from 1993 to 2009 was conducted. The initial O139 isolates were resistant to streptomycin, trimethoprim-sulfamethoxazole and polymyxin B only, while multidrug resistance increased suddenly and became common in strains isolated after 1998. Different resistance profiles were observed in the isolates from different years. In contrast, most V. cholerae O1 strains isolated in the same period were much less resistant to these antibiotics and no obvious multidrug resistance patterns were detected. Most of the non-toxigenic strains isolated from the environment and seafood were resistant to four antibiotics or fewer, although a few multidrug resistant strains were also identified. These toxigenic O139 strains exhibited a high prevalence of the class I integron and the SXT element, which were rare in the non-toxigenic strains. Molecular subtyping of O139 strains showed highly diverse pulsed-field gel electrophoresis patterns, which may correspond to the epidemic state of sporadic cases and small-scale outbreaks and complex resistance patterns. Severe multidrug resistance, even resistance transfers based on mobile antibiotic resistance elements, increases the probability of O139 cholera as a threat to public health. Therefore, continual epidemiological and antibiotic sensitivity surveillance should focus on the

  19. Multiple antibiotic resistance of Vibrio cholerae serogroup O139 in China from 1993 to 2009.

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    Li Yu

    Full Text Available Regarded as an emerging diarrheal micropathogen, Vibrio cholerae serogroup O139 was first identified in 1992 and has become an important cause of cholera epidemics over the last two decades. O139 strains have been continually isolated since O139 cholera appeared in China in 1993, from sporadic cases and dispersed foodborne outbreaks, which are the common epidemic types of O139 cholera in China. Antibiotic resistance profiles of these epidemic strains are required for development of clinical treatments, epidemiological studies and disease control. In this study, a comprehensive investigation of the antibiotic resistance of V. cholerae O139 strains isolated in China from 1993 to 2009 was conducted. The initial O139 isolates were resistant to streptomycin, trimethoprim-sulfamethoxazole and polymyxin B only, while multidrug resistance increased suddenly and became common in strains isolated after 1998. Different resistance profiles were observed in the isolates from different years. In contrast, most V. cholerae O1 strains isolated in the same period were much less resistant to these antibiotics and no obvious multidrug resistance patterns were detected. Most of the non-toxigenic strains isolated from the environment and seafood were resistant to four antibiotics or fewer, although a few multidrug resistant strains were also identified. These toxigenic O139 strains exhibited a high prevalence of the class I integron and the SXT element, which were rare in the non-toxigenic strains. Molecular subtyping of O139 strains showed highly diverse pulsed-field gel electrophoresis patterns, which may correspond to the epidemic state of sporadic cases and small-scale outbreaks and complex resistance patterns. Severe multidrug resistance, even resistance transfers based on mobile antibiotic resistance elements, increases the probability of O139 cholera as a threat to public health. Therefore, continual epidemiological and antibiotic sensitivity surveillance

  20. 泉州地区1962-2010年霍乱弧菌菌型变迁及耐药性研究%Study on variance and antibiotic resistance of Vibrio cholerae strains in Quanzhou during 1962-2010

    Institute of Scientific and Technical Information of China (English)

    李锋平; 苏培聪; 杨德林; 张庆虎

    2011-01-01

    Objective To study the epidemic isolates and antibiotic resistance of Vibrio cholera in Quanzhou,and provide the references for prevention and control of cholera. Methods Retrospective analysis was adopted based on the epidemiologjcal data on cholera in Quanzhou during 1962-2010.The antibiotic susceptibility of all the strains to antibacterials was determined by improve K-B method recommended by WHO. Results There were four cholera disease outbreaks occurred in Quanzhou city during 1962-2010. The dominant serotype that caused epidemics showed vicissitudinous phenomenon. Vibrio cholerae serotype Ogawa and Inaba were sensitive to norfloxacin and ciprofloxacin with a susceptibility of 92.31% and 99.20% , respectively .The resistance to sulphanilamide was increased year by year and the resistances to other antibiotics were high. The drug resistance of Vibrio cholerae 0139 strains was higher than that of Vibrio cholerae 01. Conclusions The dominant biotype of Vibrio cholerae was El Tor in Quanzhou city.Vibrio cholerae serotype Ogawa and Inaba that caused epidemics showed vicissitudinous phenomenon. The antibiotics sensitivity of Vibrio cholerae was gradually declining.%目的 了解泉州地区历年霍乱菌型变迁及药物耐药性,为霍乱防治工作提供参考.方法 对泉州地区1962-2010年霍乱流行疫情资料进行回顾性分析;采用WHO推荐的改良K-B纸片法,对部分菌株进行抗菌药物的药敏试验.结果 1962-2010年,泉州共发生4次较大规模的霍乱流行,流行菌型由O1小川型与O1稻叶型交替进行.大多数霍乱弧菌对诺氟沙星和环丙沙星敏感,敏感率分别为92.31%和99.20%;磺胺类药物敏感性逐年降低,对其它抗菌药耐药;O139群霍乱弧菌的耐药性明显高于O1群霍乱弧菌,不同年份的菌株耐药的程度不一致.结论 泉州地区霍乱流行优势菌型为O1群霍乱弧菌,由小川型与稻叶型交替进行;霍乱弧菌对抗菌药物的敏感性逐渐下降.

  1. O139霍乱弧菌肠毒素核苷酸序列分析%Nucleotide sequence analysis of cholera toxin in Vibrio cholerae O139

    Institute of Scientific and Technical Information of China (English)

    王军; 王玺华; 白文林; 施红; 金磊

    2000-01-01

    目的 探讨O139霍乱弧菌肠毒素(CTX)核苷酸与O1群霍乱CTX毒素核苷酸序列异同.方法 用聚合酶链反应、DNA序列分析测定2株O139群、2株O1群古典型、2株O1群E1 Tor型霍乱弧菌CTX A2-B亚单位核苷酸.结果 2株O139群霍乱弧菌均含有CTX A2-B亚单位基因,O139群与O1群CTX A2-B核苷酸同源性为97.1%~98.9%.结论 O139群与O1群霍乱弧菌CTX A2-B核苷酸基本同源.进一步证实两者CTX核苷酸序列一致.%Objective To investigate the difference of nucleotide sequence between cholera toxin (CTX)of O139 and O1 Vibrio cholerae.Methods Polymerase chain reaction and DNA sequence analysis were used to study 2 strains of O139.2 strains of classical biotype and 2 strains of EI Tor hiotype.Results Both the 2 strains of cholerae O139 contained the fragment of CTX A2-B gene,and the homology befween O1 and O139 serogroups was 98.9%~97.1%.Conclusion The nucleotide sequence of CTX A2-B in Vibrio cholerae O139 was almost consistent with that in O1,which reconfirmed the consistence of the nucleotide sequence of CTX in the 2 serogroups of Vibrio cholorae.

  2. Whole-Genome Enrichment Provides Deep Insights into Vibrio cholerae Metagenome from an African River.

    Science.gov (United States)

    Vezzulli, L; Grande, C; Tassistro, G; Brettar, I; Höfle, M G; Pereira, R P A; Mushi, D; Pallavicini, A; Vassallo, P; Pruzzo, C

    2016-11-25

    The detection and typing of Vibrio cholerae in natural aquatic environments encounter major methodological challenges related to the fact that the bacterium is often present in environmental matrices at very low abundance in nonculturable state. This study applied, for the first time to our knowledge, a whole-genome enrichment (WGE) and next-generation sequencing (NGS) approach for direct genotyping and metagenomic analysis of low abundant V. cholerae DNA (V. cholerae metagenomic DNA via hybridization. An enriched V. cholerae metagenome library was generated and sequenced on an Illumina MiSeq platform. Up to 1.8 × 10(7) bp (4.5× mean read depth) were found to map against V. cholerae reference genome sequences representing an increase of about 2500 times in target DNA coverage compared to theoretical calculations of performance for shotgun metagenomics. Analysis of metagenomic data revealed the presence of several V. cholerae virulence and virulence associated genes in river water including major virulence regions (e.g. CTX prophage and Vibrio pathogenicity island-1) and genetic markers of epidemic strains (e.g. O1-antigen biosynthesis gene cluster) that were not detectable by standard culture and molecular techniques. Overall, besides providing a powerful tool for direct genotyping of V. cholerae in complex environmental matrices, this study provides a 'proof of concept' on the methodological gap that might currently preclude a more comprehensive understanding of toxigenic V. cholerae emergence from natural aquatic environments.

  3. Naturally Occurring IgG Antibodies Provide Innate Protection against Vibrio cholerae Bacteremia by Recognition of the Outer Membrane Protein U.

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    Aung, Kyaw Min; Sjöström, Annika E; von Pawel-Rammingen, Ulrich; Riesbeck, Kristian; Uhlin, Bernt Eric; Wai, Sun Nyunt

    2016-01-01

    Cholera epidemics are caused by Vibrio cholerae serogroups O1 and O139, whereas strains collectively known as non-O1/non-O139 V. cholerae are found in cases of extraintestinal infections and bacteremia. The mechanisms and factors influencing the occurrence of bacteremia and survival of V. cholerae in normal human serum have remained unclear. We found that naturally occurring IgG recognizing V. cholerae outer membrane protein U (OmpU) mediates a serum-killing effect in a complement C1q-dependent manner. Moreover, outer membrane vesicles (OMVs) containing OmpU caused enhanced survival of highly serum-sensitive classical V. cholerae in a dose-dependent manner. OMVs from wild-type and ompU mutant V. cholerae thereby provided a novel means to verify by extracellular transcomplementation the involvement of OmpU. Our data conclusively indicate that loss, or reduced expression, of OmpU imparts resistance to V. cholerae towards serum killing. We propose that the difference in OmpU protein levels is a plausible reason for differences in serum resistance and the ability to cause bacteremia observed among V. cholerae biotypes. Our findings provide a new perspective on how naturally occurring antibodies, perhaps induced by members of the microbiome, may play a role in the recognition of pathogens and the provocation of innate immune defense against bacteremia.

  4. An in silico, in vitro and in vivo investigation of indole-3-carboxaldehyde identified from the seawater bacterium Marinomonas sp. as an anti-biofilm agent against Vibrio cholerae O1.

    Science.gov (United States)

    Rajalaxmi, Murugan; Beema Shafreen, Rajamohamed; Iyer, Prasanth M; Sahaya Vino, Raja; Balamurugan, Krishnaswamy; Pandian, Shunmugiah Karutha

    2016-01-01

    Biofilm formation is a major contributing factor in the pathogenesis of Vibrio cholerae O1 (VCO1) and therefore preventing biofilm formation could be an effective alternative strategy for controlling cholera. The present study was designed to explore seawater bacteria as a source of anti-biofilm agents against VCO1. Indole-3-carboxaldehyde (I3C) was identified as an active principle component in Marinomonas sp., which efficiently inhibited biofilm formation by VCO1 without any selection pressure. Furthermore, I3C applications also resulted in considerable collapsing of preformed pellicles. Real-time PCR studies revealed the down-regulation of virulence gene expression by modulation of the quorum-sensing pathway and enhancement of protease production, which was further confirmed by phenotypic assays. Furthermore, I3C increased the survival rate of Caenorhabditis elegans when infected with VCO1 by significantly reducing in vivo biofilm formation, which was corroborated by a survivability assay. Thus, this study revealed, for the first time, the potential of I3C as an anti-biofilm agent against VCO1.

  5. Vibrio cholerae hemolysin is required for lethality, developmental delay, and intestinal vacuolation in Caenorhabditis elegans.

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    Hediye Nese Cinar

    Full Text Available BACKGROUND: Cholera toxin (CT and toxin-co-regulated pili (TCP are the major virulence factors of Vibrio cholerae O1 and O139 strains that contribute to the pathogenesis of disease during devastating cholera pandemics. However, CT and TCP negative V. cholerae strains are still able to cause severe diarrheal disease in humans through mechanisms that are not well understood. METHODOLOGY/PRINCIPAL FINDINGS: To determine the role of other virulence factors in V. cholerae pathogenesis, we used a CT and TCP independent infection model in the nematode Caenorhabditis elegans and identified the hemolysin A (hlyA gene as a factor responsible for animal death and developmental delay. We demonstrated a correlation between the severity of infection in the nematode and the level of hemolytic activity in the V. cholerae biotypes. At the cellular level, V. cholerae infection induces formation of vacuoles in the intestinal cells in a hlyA dependent manner, consistent with the previous in vitro observations. CONCLUSIONS/SIGNIFICANCE: Our data strongly suggest that HlyA is a virulence factor in C. elegans infection leading to lethality and developmental delay presumably through intestinal cytopathic changes.

  6. Stepwise changes in viable but nonculturable Vibrio cholerae cells.

    Science.gov (United States)

    Imamura, Daisuke; Mizuno, Tamaki; Miyoshi, Shin-ichi; Shinoda, Sumio

    2015-05-01

    Many bacterial species are known to become viable but nonculturable (VBNC) under conditions that are unsuitable for growth. In this study, the requirements for resuscitation of VBNC-state Vibrio cholerae cells were found to change over time. Although VBNC cells could initially be converted to culturable by treatment with catalase or HT-29 cell extract, they subsequently entered a state that was not convertible to culturable by these factors. However, fluorescence microscopy revealed the presence of live cells in this state, from which VBNC cells were resuscitated by co-cultivation with HT-29 human colon adenocarcinoma cells. Ultimately, all cells entered a state from which they could not be resuscitated, even by co-cultivation with HT-29. These characteristic changes in VBNC-state cells were a common feature of strains in both V. cholerae O1 and O139 serogroups. Thus, the VBNC state of V. cholerae is not a single property but continues to change over time.

  7. Widespread epidemic cholera caused by a restricted subset of Vibrio cholerae clones.

    Science.gov (United States)

    Moore, S; Thomson, N; Mutreja, A; Piarroux, R

    2014-05-01

    Since 1817, seven cholera pandemics have plagued humankind. As the causative agent, Vibrio cholerae, is autochthonous in the aquatic ecosystem and some studies have revealed links between outbreaks and fluctuations in climatic and aquatic conditions, it has been widely assumed that cholera epidemics are triggered by environmental factors that promote the growth of local bacterial reservoirs. However, mounting epidemiological findings and genome sequence analysis of clinical isolates have indicated that epidemics are largely unassociated with most of the V. cholerae strains in aquatic ecosystems. Instead, only a specific subset of V. cholerae El Tor 'types' appears to be responsible for current epidemics. A recent report examining the evolution of a variety of V. cholerae strains indicates that the current pandemic is monophyletic and originated from a single ancestral clone that has spread globally in successive waves. In this review, we examine the clonal nature of the disease, with the example of the recent history of cholera in the Americas. Epidemiological data and genome sequence-based analysis of V. cholerae isolates demonstrate that the cholera epidemics of the 1990s in South America were triggered by the importation of a pathogenic V. cholerae strain that gradually spread throughout the region until local outbreaks ceased in 2001. Latin America remained almost unaffected by the disease until a new toxigenic V. cholerae clone was imported into Haiti in 2010. Overall, cholera appears to be largely caused by a subset of specific V. cholerae clones rather than by the vast diversity of V. cholerae strains in the environment.

  8. Draft Genome Sequences of Two Yersinia pseudotuberculosis ST43 (O:1b) Strains, B-7194 and B-7195.

    Science.gov (United States)

    Blouin, Yann; Platonov, Mikhail E; Pourcel, Christine; Evseeva, Vera V; Afanas'ev, Maxim V; Balakhonov, Sergey V; Anisimov, Andrey P; Vergnaud, Gilles

    2013-07-18

    We report the first draft genome sequences of two Yersinia pseudotuberculosis sequence type 43 (ST43) (O:1b) strains, B-7194 and B-7195, isolated in Russia. The total lengths of the assemblies are 4,427,121 bp and 4,608,472 bp, and 3,819 and 4,018 coding sequences, respectively, were predicted within the genomes.

  9. Antimicrobial Susceptibility of Autochthonous Aquatic Vibrio cholerae in Haiti

    Science.gov (United States)

    Baron, Sandrine; Lesne, Jean; Jouy, Eric; Larvor, Emeline; Kempf, Isabelle; Boncy, Jacques; Rebaudet, Stanilas; Piarroux, Renaud

    2016-01-01

    We investigated the antimicrobial susceptibility of 50 environmental isolates of Vibrio cholerae non-O1/non-O139 collected in surface waters in Haiti in July 2012, during an active cholera outbreak. A panel of 16 antibiotics was tested on the isolates using the disk diffusion method and PCR detection of seven resistance-associated genes (strA/B, sul1/2, ermA/B, and mefA). All isolates were susceptible to amoxicillin-clavulanic acid, cefotaxime, imipenem, ciprofloxacin, norfloxacin, amikacin, and gentamicin. Nearly a quarter (22.0%) of the isolates were susceptible to all 16 antimicrobials tested and only 8.0% of the isolates (n = 4) were multidrug-resistant. The highest proportions of resistant isolates were observed for sulfonamide (70.0%), amoxicillin (12.0%), and trimethoprim-sulfamethoxazole (10.0%). One strain was resistant to erythromycin and one to doxycycline, two antibiotics used to treat cholera in Haiti. Among the 50 isolates, 78% possessed at least two resistance-associated genes, and the genes sul1, ermA, and strB were detected in all four multidrug-resistant isolates. Our results clearly indicate that the autochthonous population of V. cholerae non-O1/non-O139 found in surface waters in Haiti shows antimicrobial patterns different from that of the outbreak strain. The presence in the Haitian aquatic environment of V. cholerae non-O1/non-O139 with reduced susceptibility or resistance to antibiotics used in human medicine may constitute a mild public health threat. PMID:27818656

  10. In vitro evaluation of capsaicin inhibitory effects on zonula occludens toxin in vibrio cholerae ATCC14035 strain

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    Soroor Erfanimanesh

    2014-10-01

    Conclusion: Capsaicin is one of the active compounds of red chili that can drastically suppress zot gene expression and shows promising inhibitory effect against V. cholerae zot production. Thus, routine intake of red chilli, which is easily available and inexpensive, may be an alternative approach to prevent and control symptoms of cholera.

  11. The Amazonia Variant of Vibrio cholerae: Molecular Identification and Study of Virulence Genes

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    Baptista MAS

    1998-01-01

    Full Text Available The pathogenic O1 Amazonia variant of Vibrio cholerae has been shown previously to have a cytotoxin acting on cultured Vero and Y-1 cells, and to lack important virulence factors such as the cholera toxin (Coelho et al. 1995a. This study extends the molecular analysis of the Amazonia strains, detecting the presence of the toxR gene, with a very similar sequence to that of the El Tor and classical biotypes. The outer membrane proteins are analyzed, detecting a variation among the group of Amazonia strains, with three different patterns found. As a by-product of this work a polymerase chain reaction fragment was sequenced, reading part of the sequence of the Lon protease of the Amazonia strains. This gene was not previously described in V. cholerae, but its sequence is present in the TIGR database specific for this species.

  12. Yersinia pseudotuberculosis ST42 (O:1) Strain Misidentified as Yersinia pestis by Mass Spectrometry Analysis.

    Science.gov (United States)

    Gérôme, Patrick; Le Flèche, Philippe; Blouin, Yann; Scholz, Holger C; Thibault, François M; Raynaud, Françoise; Vergnaud, Gilles; Pourcel, Christine

    2014-06-12

    We report here the draft sequence of strain CEB14_0017, alias HIAD_DUP, recovered from a human patient and initially identified as Yersinia pestis by mass spectrometry analysis. Genotyping based on tandem repeat polymorphism assigned the strain to Yersinia pseudotuberculosis sequence type 42 (ST42). The total assembly length is 4,894,739 bp.

  13. Detection of Vibrio cholerae by isothermal cross-priming amplification combined with nucleic acid detection strip analysis.

    Science.gov (United States)

    Zhang, Xia; Du, Xin-Jun; Guan, Chun; Li, Ping; Zheng, Wen-Jie; Wang, Shuo

    2015-08-01

    Vibrio cholerae is a water- and food-borne human pathogen, and V. cholerae serotypes O1 and O139 have attracted attention because of their severe pathogenesis. However, non-O1, non-O139 cholera vibrios (NCVs) were also recently recognized as having virulence properties. In this study, we developed a cross-priming amplification (CPA) method for the detection of all serotypes of V. cholerae. The specificity of the CPA method was tested using a panel of 60 different bacterial strains. All of the V. cholerae strains showed positive results, and 41 other types of bacteria gave negative results. The limit of detection of the CPA method was 79.28 fg of genomic DNA, 4.2 × 10(2) CFU/ml for bacteria in pure culture, and 5.6 CFU per 25 g of sample with pre-enrichment. This method showed a higher sensitivity than the loop-mediated isothermal amplification (LAMP) method did and was more convenient to perform. These results indicate that the CPA method can be used for the rapid preliminary screening of V. cholerae.

  14. Fish as Hosts of Vibrio cholerae

    Science.gov (United States)

    Halpern, Malka; Izhaki, Ido

    2017-01-01

    Vibrio cholerae, the causative agent of pandemic cholera, is abundant in marine and freshwater environments. Copepods and chironomids are natural reservoirs of this species. However, the ways V. cholerae is globally disseminated are as yet unknown. Here we review the scientific literature that provides evidence for the possibility that some fish species may be reservoirs and vectors of V. cholerae. So far, V. cholerae has been isolated from 30 fish species (22 freshwater; 9 marine). V. cholerae O1 was reported in a few cases. In most cases V. cholerae was isolated from fish intestines, but it has also been detected in gills, skin, kidney, liver and brain tissue. In most cases the fish were healthy but in some, they were diseased. Nevertheless, Koch postulates were not applied to prove that V. cholerae and not another agent was the cause of the disease in the fish. Evidence from the literature correlates raw fish consumption or fish handling to a few cholera cases or cholera epidemics. Thus, we can conclude that V. cholerae inhabits some marine and freshwater fish species. It is possible that fish may protect the bacteria in unfavorable habitats while the bacteria may assist the fish to digest its food. Also, fish may disseminate the bacteria in the aquatic environment and may transfer it to waterbirds that consume them. Thus, fish are reservoirs of V. cholerae and may play a role in its global dissemination. PMID:28293221

  15. Vibrio cholerae and Vibrio parahaemolyticus detected in seafood products from Senegal.

    Science.gov (United States)

    Coly, Ignace; Sow, Amy Gassama; Seydi, Malang; Martinez-Urtaza, Jaime

    2013-12-01

    The detection of pathogenic Vibrio in seafood from Senegal has generated five food alerts in the European Union. To investigate the presence and abundance Vibrio cholerae and Vibrio parahaemolyticus in seafood and coastal and estuarine waters, 123 seafood samples and 52 water samples were collected during 2007-2009 from two large seafood markets in Dakar, and from different oceanic and estuarine areas of the country. V. parahaemolyticus was detected in 30.1% of seafood samples, whereas presence of V. cholerae was only found in 1.6%. In water samples, V. parahaemolyticus and V. cholerae were detected in 28.8% and 5.7% of the samples, respectively. Abundance of V. parahaemolyticus in seafood from the fishing areas ranged from 110 MPN/g. Densities of V. cholerae in the two positive seafood samples reached values of 0.36 and 0.61 MPN/g, repectively. V. parahaemolyticus strains were found to possess tlh, but not tdh and trh by polymerase chain reaction, and all the strains of V. cholerae were non-O1 or non-O139. These results suggest that the prevalence of high salinities in coastal and estuarine environments of Senegal limits the occurrence of V. parahaemolyticus and V. cholerae, despite warmer temperatures prevailing in seawater environments throughout the year. Furthermore, temperature abuse driven by a deficient cold chain over the distribution and retail sales may represent a major risk due to the postharvest multiplication of these Vibrio pathogens.

  16. [ACTUAL PROBLEMS OF EPIDEMIOLOGIC CONTROL, LABORATORY DIAGNOSTICS AND PROPHYLAXIS OF CHOLERA IN RUSSIAN FEDERATION].

    Science.gov (United States)

    Onischenko, G G; Popova, A Yu; Kutyrev, V V; Smirnova, N I; Scherbakova, S A; Moskvitina, E A; Titova, S V

    2016-01-01

    Main problems of system of epidemiologic control for cholera active in Russian Federation, as well as laboratory diagnostics and vaccine prophylaxis of this especially dangerous infection, that had emerged in the contemporary period of the ongoing 7th pandemic of cholera, are discussed. Features of the genome of natural strains of Vibrio cholerae of El Tor biovar, that possess a poten- tial epidemic threat, as well as problems, that have emerged during isolation of these strains from samples of water of surface water bodies during their monitoring, are also examined. The main direction of enhancement of the system of epidemiologic control for cholera consist in develop- ment of a new algorithm of differentiation of administrative territories of Russian Federation by types of epidemic manifestations, as well as optimization of monitoring of environment objects. Integration of modern highly informative technologies into practice, as well as development of new generation diagnostic preparations based on DNA-chips and immunechips is necessary to increase effectiveness of the conducted operative and retrospective diagnostics in the contemporary period. Creation of national cholera vaccine, ensuring simultaneous protection from cholera causative agents of both O1 and O139 serogroups, is also required.

  17. Virulence Gene PCR and PFGE Genotyping analysis of Vibrio cholerae strains isolated from cholera epidemics in Hunan province from 2005 to 2010%湖南省2005年-2010年霍乱疫情分离株的毒力基因PCR及PFGE分型分析

    Institute of Scientific and Technical Information of China (English)

    夏昕; 湛志飞; 覃迪; 刘运芝; 高立冬; 胡世雄; 邓志红; 张红

    2012-01-01

    Objective; To understand the pathogenic characteristics of Vibrio cholerae 0139 strains isolated from Vibrio cholerae epidemics in Hunan province from 2005 to 2010; to study the colone relations among the strains. Methods: K - B method was employed to test drug sensitivity; ctxAB virulence gene was tested by PCR, and finally molecular typing was carried out by pulsed field gel electrophoresis ( PFGE) for representative strains isolated from Vibrio cholerae epidemics. Results; 33 Vibrio cholerae 0139 stains presented a higher drug resistance rate against doxycycline and sulphame -thoxazole of 39. 39% and 75.76% , while a sensitivity of 100% to ciprofloxacin, nor-floxacin and amikacin; The virulence gene PCR results showed all the Vibrio cholerae 0139 strains were cholera toxin genes ctxAB - positive; 24 Vibrio cholerae 0139 strains isolated from Vibrio cholerae epidemics in 2005 and 2010 showed 3 PFGE banding types,and all the strains were homology of 83% - 100% by cluster analysis. Conclusion; Vibrio cholerae 0139 strains isolated from cholera epidemic in Hunan province from 2005 to 2010 were all ctxAB positive. The strains from different years and regions were found the closely related epidemic clone group strains of cholera; Resistance monitoring and further molecular typing analysis of Cholera strains contribute to the efficient surveillance of cholera and infectious source tracking.%目的:了解2005年-2010年湖南省霍乱疫情分离到的O139群霍乱弧菌菌株的病原学特征,研究疫情分离株之间的克隆相关性.方法:采用K-B法进行药敏试验;聚合酶链反应(PCR)检测ctxAB毒力基因;脉冲场凝胶电泳对疫情分离代表株进行PFGE分型分析.结果:33株霍乱弧菌对强力霉素、复方新诺明的耐药率较高,分别为39.39%和75.76%,对环丙沙星、诺氟沙星以及丁胺卡那100%敏感;毒力基因的PCR结果显示为所有疫情分离的O139霍乱弧菌均为产毒株,即

  18. Molecular epidemiology of Vibrio cholerae associated with flood in Brahamputra River valley, Assam, India.

    Science.gov (United States)

    Bhuyan, Soubhagya K; Vairale, Mohan G; Arya, Neha; Yadav, Priti; Veer, Vijay; Singh, Lokendra; Yadava, Pramod K; Kumar, Pramod

    2016-06-01

    Cholera is often caused when drinking water is contaminated through environmental sources. In recent years, the drastic cholera epidemics in Odisha (2007) and Haiti (2010) were associated with natural disasters (flood and Earthquake). Almost every year the state of Assam India witnesses flood in Brahamputra River valley during reversal of wind system (monsoon). This is often followed by outbreak of diarrheal diseases including cholera. Beside the incidence of cholera outbreaks, there is lack of experimental evidence for prevalence of the bacterium in aquatic environment and its association with cholera during/after flood in the state. A molecular surveillance during 2012-14 was carried out to study prevalence, strain differentiation, and clonality of Vibrio cholerae in inland aquatic reservoirs flooded by Brahamputra River in Assam. Water samples were collected, filtered, enriched in alkaline peptone water followed by selective culturing on thiosulfate bile salt sucrose agar. Environmental isolates were identified as V. cholerae, based on biochemical assays followed by sero-grouping and detailed molecular characterization. The incidence of the presence of the bacterium in potable water sources was higher after flood. Except one O1 isolate, all of the strains were broadly grouped under non-O1/non-O139 whereas some of them did have cholera toxin (CT). Surprisingly, we have noticed Haitian ctxB in two non-O1/non-O139 strains. MLST analyses based on pyrH, recA and rpoA genes revealed clonality in the environmental strains. The isolates showed varying degree of antimicrobial resistance including tetracycline and ciprofloxacin. The strains harbored the genetic elements SXT constins and integrons responsible for multidrug resistance. Genetic characterization is useful as phenotypic characters alone have proven to be unsatisfactory for strain discrimination. An assurance to safe drinking water, sanitation and monitoring of the aquatic reservoirs is of utmost importance for

  19. Cholera outbreaks in Africa.

    Science.gov (United States)

    Mengel, Martin A; Delrieu, Isabelle; Heyerdahl, Leonard; Gessner, Bradford D

    2014-01-01

    an infectious dose of Vibrio cholerae and on the virulence of the implicated strain. Cholera transmission can then be amplified by several factors including contamination of human water- or food sources; climate and extreme weather events; political and economic crises; high population density combined with poor quality informal housing and poor hygiene practices; spread beyond a local community through human travel and animals, e.g., water birds. At an individual level, cholera risk may increase with decreasing immunity and hypochlorhydria, such as that induced by Helicobacter pylori infection, which is endemic in much of Africa, and may increase individual susceptibility and cholera incidence. Since contaminated water is the main vehicle for the spread of cholera, the obvious long-term solution to eradicate the disease is the provision of safe water to all African populations. This requires considerable human and financial resources and time. In the short and medium term, vaccination may help to prevent and control the spread of cholera outbreaks. Regardless of the intervention, further understanding of cholera biology and epidemiology is essential to identify populations and areas at increased risk and thus ensure the most efficient use of scarce resources for the prevention and control of cholera.

  20. Cholera with severe renal failure in an Italian tourist returning from Cuba, July 2013.

    Science.gov (United States)

    Mascarello, M; Deiana, M L; Maurel, C; Lucarelli, C; Luzzi, I; Luzzati, R

    2013-08-29

    In July 2013, an Italian tourist returning from Cuba was hospitalised in Trieste, Italy, for cholera caused by Vibrio cholerae O1 serotype Ogawa with severe renal failure. An outbreak of cholera was reported in Cuba in January 2013. Physicians should consider the diagnosis of cholera in travellers returning from Cuba presenting with acute watery diarrhoea.

  1. Rapid detection by multiplex PCR of Genomic Islands, prophages and Integrative Conjugative Elements in V. cholerae 7th pandemic variants.

    Science.gov (United States)

    Spagnoletti, Matteo; Ceccarelli, Daniela; Colombo, Mauro M

    2012-01-01

    Vibrio cholerae poses a threat to human health, and new epidemic variants have been reported so far. Seventh pandemic V. cholerae strains are characterized by highly related genomic sequences but can be discriminated by a large set of Genomic Islands, phages and Integrative Conjugative Elements. Classical serotyping and biotyping methods do not easily discriminate among new variants arising worldwide, therefore the establishment of new methods for their identification is required. We developed a multiplex PCR assay for the rapid detection of the major 7th pandemic variants of V. cholerae O1 and O139. Three specific genomic islands (GI-12, GI-14 and GI-15), two phages (Kappa and TLC), Vibrio Seventh Pandemic Island 2 (VSP-II), and the ICEs of the SXT/R391 family were selected as targets of our multiplex PCR based on a comparative genomic approach. The optimization and specificity of the multiplex PCR was assessed on 5 V. cholerae 7th pandemic reference strains, and other 34 V. cholerae strains from various epidemic events were analyzed to validate the reliability of our method. This assay had sufficient specificity to identify twelve different V. cholerae genetic profiles, and therefore has the potential to be used as a rapid screening method.

  2. Drinking cholera

    DEFF Research Database (Denmark)

    Grant, Stephen Lawrence; Tamason, Charlotte Crim; Hoque, Bilqis Amin

    2015-01-01

    beconducive to V. cholerae survival. Furthermore, salinity levels of participant’s drinking water sourceswere all well below the levels required for optimal survival of V. cholerae. Respondents explainedthat they preferred less salty and more aesthetically pleasing drinking water. Conclusion: Theoretically, V...

  3. Induction of immunomodulatory miR-146a and miR-155 in small intestinal epithelium of Vibrio cholerae infected patients at acute stage of cholera

    Science.gov (United States)

    Melgar, Silvia; Aung, Kyaw Min; Rahman, Arman; Qadri, Firdausi; Wai, Sun Nyunt; Shirin, Tahmina

    2017-01-01

    The potential immunomodulatory role of microRNAs in small intestine of patients with acute watery diarrhea caused by Vibrio cholerae O1 or enterotoxigenic Escherichia coli (ETEC) infection was investigated. Duodenal biopsies were obtained from study-participants at the acute (day 2) and convalescent (day 21) stages of disease, and from healthy individuals. Levels of miR-146a, miR-155 and miR-375 and target gene (IRAK1, TRAF6, CARD10) and 11 cytokine mRNAs were determined by qRT-PCR. The cellular source of microRNAs in biopsies was analyzed by in situ hybridization. The ability of V. cholerae bacteria and their secreted products to cause changes in microRNA- and mRNA levels in polarized tight monolayers of intestinal epithelial cells was investigated. miR-146a and miR-155 were expressed at significantly elevated levels at acute stage of V. cholerae infection and declined to normal at convalescent stage (Pcholerae O1 strain C6706 and clinical isolates from two study-participants, caused significant increase in miR-155 and miR-146a by the strain C6706 (Pcholera. The inducer is probably the V. cholerae bacterium. By inducing microRNAs the bacterium can limit the innate immune response of the host, including inflammation evoked by its own secreted factors, thereby decreasing the risk of being eliminated. PMID:28319200

  4. Development of a PCR-restriction fragment length polymorphism assay for detection and subtyping of cholix toxin variant genes of Vibrio cholerae.

    Science.gov (United States)

    Awasthi, Sharda Prasad; Asakura, Masahiro; Neogi, Sucharit Basu; Hinenoya, Atsushi; Ramamurthy, T; Yamasaki, Shinji

    2014-05-01

    Cholix toxin (ChxA) is an exotoxin reported in Vibrio cholerae non-O1/non-O139. Apart from its prototype (ChxA I) we have recently identified two novel variants of this toxin, ChxA II and ChxA III. Our previous investigations indicated that the first two variants may instigate extra-intestinal infections and ChxA II can be more lethal than ChxA I in mice. However, all three cholix toxins (ChxA I to III) failed to show any enterotoxicity in rabbit ileal loops. In this study we developed a PCR-restriction fragment length polymorphism (RFLP) assay to differentiate all three chxA variants to further understand the importance of each subtype. By using 53 V. cholerae non-O1/non-O139 strains harbouring chxA genes, which were previously categorized by sequencing, and various other strains as negative controls, the PCR-RFLP assay showed 100 % typability and specificity. Furthermore, when applied to differentiate additional V. cholerae strains, which were also screened for the chxA gene by colony hybridization, this assay identified chxA I and chxA II genes among 18.5 % and 4.5 % of non-O1/non-O139 strains (n = 178), respectively. One non-O1/non-O139 strain was untypable due to the insertion of an IS911-like element. Interestingly, the chxA I gene was detected in 10 out of 137 cholera toxin gene-negative V. cholerae O1 strains. These results suggest that the PCR-RFLP assay developed in this study can be a rapid and simple method to differentiate the chxA subtypes.

  5. Structural and functional importance of outer membrane proteins in Vibrio cholerae flagellum.

    Science.gov (United States)

    Bari, Wasimul; Lee, Kang-Mu; Yoon, Sang Sun

    2012-08-01

    Vibrio cholerae has a sheath-covered monotrichous flagellum that is known to contribute to virulence. Although the structural organization of the V. cholerae flagellum has been extensively studied, the involvement of outer membrane proteins as integral components in the flagellum still remains elusive. Here we show that flagella produced by V. cholerae O1 El Tor strain C6706 were two times thicker than those from two other Gram-negative bacteria. A C6706 mutant strain (SSY11) devoid of two outer membrane proteins (OMPs), OmpU and OmpT, produced thinner flagella. SSY11 showed significant defects in the flagella-mediated motility as compared to its parental strain. Moreover, increased shedding of the flagella-associated proteins was observed in the culture supernatant of SSY11. This finding was also supported by the observation that culture supernatants of the SSY11 strain induced the production of a significantly higher level of IL-8 in human colon carcinoma HT29 and alveolar epithelial A549 cells than those of the wild-type C6706 strain. These results further suggest a definite role of these two OMPs in providing the structural integrity of the V. cholerae flagellum as part of the surrounding sheath.

  6. Targeting aphA : a new high-throughput screening assay identifies compounds that reduce prime virulence factors of Vibrio cholerae.

    Science.gov (United States)

    Bolger, Galina; Roy, Sambit; Zapol'skii, Viktor A; Kaufmann, Dieter E; Schnürch, Michael; Mihovilovic, Marko D; Nandy, Ranjan K; Tegge, Werner

    2016-07-01

    A high-throughput screening (HTS) assay was developed for identifying compounds with inhibitory effect on aphA, one of the key regulators positively controlling Vibrio cholerae pathogenesis. An inhibitory effect on aphA was expected to lead to attenuation in the secretion of the major pathogenicity factors of V. cholerae, cholera toxin and toxin co-regulated pilus. The plasmid construct pAKSB was developed with a kanamycin resistance (KmR) gene under the control of the aphA -like promoter for conferring a KmR phenotype under aphA -expressing conditions. The HTS assay was performed to identify compounds with inhibitory effect on the growth of O139 V. cholerae MO10 carrying the construct pAKSB in growth medium containing Km (30 g ml-1), but not in its absence. Of 20 338 compounds screened, six compounds were identified to inhibit the pAKSB-induced KmR phenotype and these compounds caused transcriptional inhibition of aphA in V. cholerae O139 strain MO10 as well as variant V. cholerae O1 El Tor strain NM06-058. Of the three most active substances, compound 53760866 showed lowest half-maximal cytotoxicity in a eukaryotic cell viability assay and was characterized further. Compound 53760866 caused reduction in cholera toxin secretion and expression of TcpA in vitro. The in vitro virulence attenuation corroborated well in a suckling mouse model in vivo, which showed reduction of colonization by V. cholerae NM06-058 when co-administered with 53760866. The screening method and the compounds may lead to new preventive strategies for cholera by reducing the pathogenicity of V. cholerae .

  7. Randomized, controlled human challenge study of the safety, immunogenicity, and protective efficacy of a single dose of Peru-15, a live attenuated oral cholera vaccine.

    Science.gov (United States)

    Cohen, Mitchell B; Giannella, Ralph A; Bean, Judy; Taylor, David N; Parker, Susan; Hoeper, Amy; Wowk, Stephen; Hawkins, Jennifer; Kochi, Sims K; Schiff, Gilbert; Killeen, Kevin P

    2002-04-01

    Peru-15 is a live attenuated oral vaccine derived from a Vibrio cholerae O1 El Tor Inaba strain by a series of deletions and modifications, including deletion of the entire CT genetic element. Peru-15 is also a stable, motility-defective strain and is unable to recombine with homologous DNA. We wished to determine whether a single oral dose of Peru-15 was safe and immunogenic and whether it would provide significant protection against moderate and severe diarrhea in a randomized, double-blind, placebo-controlled human volunteer cholera challenge model. A total of 59 volunteers were randomly allocated to groups to receive either 2 x 10(8) CFU of reconstituted, lyophilized Peru-15 vaccine diluted in CeraVacx buffer or placebo (CeraVacx buffer alone). Approximately 3 months after vaccination, 36 of these volunteers were challenged with approximately 10(5) CFU of virulent V. cholerae O1 El Tor Inaba strain N16961, prepared from a standardized frozen inoculum. Among vaccinees, 98% showed at least a fourfold increase in vibriocidal antibody titers. After challenge, 5 (42%) of the 12 placebo recipients and none (0%) of the 24 vaccinees had moderate or severe diarrhea (> or = 3,000 g of diarrheal stool) (P = 0.002; protective efficacy, 100%; lower one-sided 95% confidence limit, 75%). A total of 7 (58%) of the 12 placebo recipients and 1 (4%) of the 24 vaccinees had any diarrhea (P Peru-15 is a well-tolerated and immunogenic oral cholera vaccine that affords protective efficacy against life-threatening cholera diarrhea in a human volunteer challenge model. This vaccine may therefore be a safe and effective tool to prevent cholera in travelers and is a strong candidate for further evaluation to prevent cholera in an area where cholera is endemic.

  8. Promotion of colonization and virulence by cholera toxin is dependent on neutrophils.

    Science.gov (United States)

    Queen, Jessica; Satchell, Karla J F

    2013-09-01

    The innate immune response to Vibrio cholerae infection is poorly understood, but this knowledge is critical for the design of safe, effective vaccines. Using an adult mouse intestinal infection model, this study examines the contribution of neutrophils to host immunity, as well as the effect of cholera toxin and other secreted factors on this response. Depletion of neutrophils from mice with anti-Ly6G IA8 monoclonal antibody led to similar survival rates of mice infected with low or moderate doses of toxigenic V. cholerae El Tor O1. At a high dose, neutropenic mice showed increased rates of survival compared to neutrophil-replete animals. Expression of cholera toxin was found to be protective to the neutropenic host, and this phenotype can be replicated by the administration of purified toxin. Neutrophils do not effectively clear colonizing bacteria from the small intestine, nor do they alter induction of early immune-modulating signals. In both neutropenic and neutrophil-replete animals, the local response to infection is characterized by expression of interleukin 6 (IL-6), IL-10, and macrophage inflammatory protein 2 alpha (MIP-2). Overall, these data indicate that the innate immune response to toxigenic V. cholerae infection differs dramatically from the host response to nontoxigenic infection or vaccination, where neutrophils are protective to the host. In the absence of neutrophils, cholera toxin induces immunomodulatory effects that increase host survival. In cholera toxin-producing strains, similar to nontoxigenic infection, accessory toxins are critical to virulence, indicating that cholera toxin and the other secreted toxins modulate the host response by different mechanisms, with both contributing to bacterial persistence and virulence.

  9. Transmission of Infectious Vibrio cholerae through Drinking Water among the Household Contacts of Cholera Patients (CHoBI7 Trial)

    Science.gov (United States)

    Rafique, Raisa; Rashid, Mahamud-ur; Monira, Shirajum; Rahman, Zillur; Mahmud, Md. Toslim; Mustafiz, Munshi; Saif-Ur-Rahman, K. M.; Johura, Fatema-Tuz; Islam, Saiful; Parvin, Tahmina; Bhuyian, Md. Sazzadul I.; Sharif, Mohsena B.; Rahman, Sabita R.; Sack, David A.; Sack, R. Bradley; George, Christine M.; Alam, Munirul

    2016-01-01

    Recurrent cholera causes significant morbidity and mortality among the growing population of Dhaka, the capital city of Bangladesh. Previous studies have demonstrated that household contacts of cholera patients are at >100 times higher risk of cholera during the week after the presentation of the index patient. Our prospective study investigated the mode of transmission of Vibrio cholerae, the cause of cholera, in the households of cholera patients in Dhaka city. Out of the total 420 rectal swab samples analyzed from 84 household contacts and 330 water samples collected from 33 households, V. cholerae was isolated from 20%(17/84) of household contacts, 18%(6/33) of stored drinking water, and 27%(9/33) of source water samples. Phenotypic and molecular analyses results confirmed the V. cholerae isolates to be toxigenic and belonging to serogroup O1 biotype El Tor (ET) possessing cholera toxin of classical biotype (altered ET). Phylogenetic analysis by pulsed-field gel electrophoresis (PFGE) showed the V. cholerae isolates to be clonally linked, as >95% similarity was confirmed by sub-clustering patterns in the PFGE (NotI)-based dendrogram. Mapping results showed cholera patients to be widely distributed across 25 police stations. The data suggesting the transmission of infectious V. cholerae within the household contacts of cholera patients through drinking water underscores the need for safe water to prevent spread of cholera and related deaths in Dhaka city. PMID:27803695

  10. Transmission of Infectious Vibrio cholerae Through Drinking Water among the Household Contacts of Cholera Patients (CHoBI7 Trial

    Directory of Open Access Journals (Sweden)

    Raisa Rafique

    2016-10-01

    Full Text Available Recurrent cholera causes significant morbidity and mortality among the growing population of Dhaka, the capital city of Bangladesh. Previous studies have demonstrated that household contacts of cholera patients are at >100 times higher risk of cholera during the week after the presentation of the index patient. Our prospective study investigated the mode of transmission of Vibrio cholerae, the cause of cholera, in the households of cholera patients in Dhaka city. Of total 420 rectal swab samples analyzed from 84 household contacts and 330 water samples collected from 33 households, V. cholerae was isolated from 20%(17/84 of household contacts, 18%(6/33 of stored drinking water, and 27%(9/33 of source water samples. Phenotypic and molecular analyses results confirmed the V. cholerae isolates to be toxigenic and belonging to serogroup O1 biotype El Tor (ET possessing cholera toxin of classical biotype (altered ET. Phylogenetic analysis by pulsed-field gel electrophoresis (PFGE showed the V. cholerae isolates to be clonally linked, as >95% similarity was confirmed by sub-clustering patterns in the PFGE (NotI-based dendrogram. Mapping results showed cholera patients to be widely distributed across 25 police stations with the highest incidence in households near the major rivers and polluted water bodies. The data presented on the transmission of infectious V. cholerae within the household contacts of cholera patients through drinking water underscores the need for safe water to prevent spread of cholera and related deaths in Dhaka city.

  11. Dynamics in genome evolution of Vibrio cholerae.

    Science.gov (United States)

    Banerjee, Rachana; Das, Bhabatosh; Balakrish Nair, G; Basak, Surajit

    2014-04-01

    Vibrio cholerae, the etiological agent of the acute secretary diarrheal disease cholera, is still a major public health concern in developing countries. In former centuries cholera was a permanent threat even to the highly developed populations of Europe, North America, and the northern part of Asia. Extensive studies on the cholera bug over more than a century have made significant advances in our understanding of the disease and ways of treating patients. V. cholerae has more than 200 serogroups, but only few serogroups have caused disease on a worldwide scale. Until the present, the evolutionary relationship of these pandemic causing serogroups was not clear. In the last decades, we have witnessed a shift involving genetically and phenotypically varied pandemic clones of V. cholerae in Asia and Africa. The exponential knowledge on the genome of several representatives V. cholerae strains has been used to identify and analyze the key determinants for rapid evolution of cholera pathogen. Recent comparative genomic studies have identified the presence of various integrative mobile genetic elements (IMGEs) in V. cholerae genome, which can be used as a marker of differentiation of all seventh pandemic clones with very similar core genome. This review attempts to bring together some of the important researches in recent times that have contributed towards understanding the genetics, epidemiology and evolution of toxigenic V. cholerae strains.

  12. Successful small intestine colonization of adult mice by Vibrio cholerae requires ketamine anesthesia and accessory toxins.

    Directory of Open Access Journals (Sweden)

    Verena Olivier

    Full Text Available Vibrio cholerae colonizes the small intestine of adult C57BL/6 mice. In this study, the physical and genetic parameters that facilitate this colonization were investigated. Successful colonization was found to depend upon anesthesia with ketamine-xylazine and neutralization of stomach acid with sodium bicarbonate, but not streptomycin treatment. A variety of common mouse strains were colonized by O1, O139, and non-O1/non-O139 strains. All combinations of mutants in the genes for hemolysin, the multifunctional, autoprocessing RTX toxin (MARTX, and hemagglutinin/protease were assessed, and it was found that hemolysin and MARTX are each sufficient for colonization after a low dose infection. Overall, this study suggests that, after intragastric inoculation, V. cholerae encounters barriers to infection including an acidic environment and an immediate immune response that is circumvented by sodium bicarbonate and the anti-inflammatory effects of ketamine-xylazine. After initial adherence in the small intestine, the bacteria are subjected to additional clearance mechanisms that are evaded by the independent toxic action of hemolysin or MARTX. Once colonization is established, it is suggested that, in humans, these now persisting bacteria initiate synthesis of the major virulence factors to cause cholera disease. This adult mouse model of intestinal V. cholerae infection, now well-characterized and fully optimized, should serve as a valuable tool for studies of pathogenesis and testing vaccine efficacy.

  13. Vibrio cholerae Serogroup O139: Isolation from Cholera Patients and Asymptomatic Household Family Members in Bangladesh between 2013 and 2014.

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    Fahima Chowdhury

    2015-11-01

    Full Text Available Cholera is endemic in Bangladesh, with outbreaks reported annually. Currently, the majority of epidemic cholera reported globally is El Tor biotype Vibrio cholerae isolates of the serogroup O1. However, in Bangladesh, outbreaks attributed to V. cholerae serogroup O139 isolates, which fall within the same phylogenetic lineage as the O1 serogroup isolates, were seen between 1992 and 1993 and in 2002 to 2005. Since then, V. cholerae serogroup O139 has only been sporadically isolated in Bangladesh and is now rarely isolated elsewhere.Here, we present case histories of four cholera patients infected with V. cholerae serogroup O139 in 2013 and 2014 in Bangladesh. We comprehensively typed these isolates using conventional approaches, as well as by whole genome sequencing. Phenotypic typing and PCR confirmed all four isolates belonging to the O139 serogroup.Whole genome sequencing revealed that three of the isolates were phylogenetically closely related to previously sequenced El Tor biotype, pandemic 7, toxigenic V. cholerae O139 isolates originating from Bangladesh and elsewhere. The fourth isolate was a non-toxigenic V. cholerae that, by conventional approaches, typed as O139 serogroup but was genetically divergent from previously sequenced pandemic 7 V. cholerae lineages belonging to the O139 or O1 serogroups.These results suggest that previously observed lineages of V. cholerae O139 persist in Bangladesh and can cause clinical disease and that a novel disease-causing non-toxigenic O139 isolate also occurs.

  14. Vibrio cholerae Serogroup O139: Isolation from Cholera Patients and Asymptomatic Household Family Members in Bangladesh between 2013 and 2014

    Science.gov (United States)

    Chowdhury, Fahima; Mather, Alison E.; Begum, Yasmin Ara; Asaduzzaman, Muhammad; Baby, Nabilah; Sharmin, Salma; Biswas, Rajib; Ikhtear Uddin, Muhammad; LaRocque, Regina C.; Harris, Jason B.; Calderwood, Stephen B.; Ryan, Edward T.; Clemens, John D.; Thomson, Nicholas R.; Qadri, Firdausi

    2015-01-01

    Background Cholera is endemic in Bangladesh, with outbreaks reported annually. Currently, the majority of epidemic cholera reported globally is El Tor biotype Vibrio cholerae isolates of the serogroup O1. However, in Bangladesh, outbreaks attributed to V. cholerae serogroup O139 isolates, which fall within the same phylogenetic lineage as the O1 serogroup isolates, were seen between 1992 and 1993 and in 2002 to 2005. Since then, V. cholerae serogroup O139 has only been sporadically isolated in Bangladesh and is now rarely isolated elsewhere. Methods Here, we present case histories of four cholera patients infected with V. cholerae serogroup O139 in 2013 and 2014 in Bangladesh. We comprehensively typed these isolates using conventional approaches, as well as by whole genome sequencing. Phenotypic typing and PCR confirmed all four isolates belonging to the O139 serogroup. Findings Whole genome sequencing revealed that three of the isolates were phylogenetically closely related to previously sequenced El Tor biotype, pandemic 7, toxigenic V. cholerae O139 isolates originating from Bangladesh and elsewhere. The fourth isolate was a non-toxigenic V. cholerae that, by conventional approaches, typed as O139 serogroup but was genetically divergent from previously sequenced pandemic 7 V. cholerae lineages belonging to the O139 or O1 serogroups. Conclusion These results suggest that previously observed lineages of V. cholerae O139 persist in Bangladesh and can cause clinical disease and that a novel disease-causing non-toxigenic O139 isolate also occurs. PMID:26562418

  15. Deciphering the origin of the 2012 cholera epidemic in Guinea by integrating epidemiological and molecular analyses.

    Directory of Open Access Journals (Sweden)

    Stanislas Rebaudet

    2014-06-01

    Full Text Available Cholera is typically considered endemic in West Africa, especially in the Republic of Guinea. However, a three-year lull period was observed from 2009 to 2011, before a new epidemic struck the country in 2012, which was officially responsible for 7,350 suspected cases and 133 deaths. To determine whether cholera re-emerged from the aquatic environment or was rather imported due to human migration, a comprehensive epidemiological and molecular survey was conducted. A spatiotemporal analysis of the national case databases established Kaback Island, located off the southern coast of Guinea, as the initial focus of the epidemic in early February. According to the field investigations, the index case was found to be a fisherman who had recently arrived from a coastal district of neighboring Sierra Leone, where a cholera outbreak had recently occurred. MLVA-based genotype mapping of 38 clinical Vibrio cholerae O1 El Tor isolates sampled throughout the epidemic demonstrated a progressive genetic diversification of the strains from a single genotype isolated on Kaback Island in February, which correlated with spatial epidemic spread. Whole-genome sequencing characterized this strain as an "atypical" El Tor variant. Furthermore, genome-wide SNP-based phylogeny analysis grouped the Guinean strain into a new clade of the third wave of the seventh pandemic, distinct from previously analyzed African strains and directly related to a Bangladeshi isolate. Overall, these results highly suggest that the Guinean 2012 epidemic was caused by a V. cholerae clone that was likely imported from Sierra Leone by an infected individual. These results indicate the importance of promoting the cross-border identification and surveillance of mobile and vulnerable populations, including fishermen, to prevent, detect and control future epidemics in the region. Comprehensive epidemiological investigations should be expanded to better understand cholera dynamics and improve

  16. 应用PFGE分析广州稻叶型霍乱弧菌的同源性%Homology analysis of strains of Vibrio cholerae serotype Inaba isolated in Guangzhou by pulsed-field gel electrophoresis analysis (PFGE)

    Institute of Scientific and Technical Information of China (English)

    李孝权; 庞杏林; 张欣强; 邓志爱; 李美霞; 陈守义; 邓小玲; 王鸣

    2011-01-01

    Objective To analyze the characteristics of pathogenicity-associated genes and PFGE features of 40 strains of Vibrio cholerae (V.cholerae)serotype Inaba isolated in Guangzhou,and study homology and features of cholera epidemic caused by V.cholerae. Methods Polymerase Chain Reaction (PCR)assay was utilized to detect the four pathogenicity-associated genes of these isolates. PFGE using restriction enzyme Not I was employed in the molecular subtyping of V.cholerae isolates and PFGE patterns were analyzed by BioNumerics Version 4.0 software to perform cluster analysis and homology analysis. Pattem profiles were compared by utilizing the Dice coefficient and UPGMA (unweighted pair group method with arithmetic averages). Results Genotype A strains were found in most of the epidemic involved V. cholerae serotype Inaba. Most of environmentally isolated strains belonged to genotype C and genotype B. Eighteen distinct pulsotypes were distributed among 40 isolates typeable by PFGE. Closely related pulsotypes existed among V. cholerae isolates from cholera epidemic(with similarity value above 98% ) and pulsotypes of environmental strains demonstrated obvious diversity. Conclusions One of the major differences of V. cholerae serotype lnaba from cholera epidemic and the environment is whether they carry ctxA gene. Relative low homology was observed among V.cholerae serotype lnaba strains from these two sources, but it is still necessary to enforce the homology analysis and track the source of V. cholerae.%目的 分析广州40株稻叶型霍乱弧菌的毒力基因和PFGE特征,探讨菌株的同源性和广州稻叶型霍乱弧菌流行的特点.方法 应用PCR方法 检测稻叶型菌株的4种毒力基因,采用限制性内切酶Not I对菌株进行PFGE分型,使用BioNumerics Version 4.0软件(复选Dice相关系数和UPGMA方法 )对菌株进行聚类和同源分析.结果 绝大多数水型稻叶霍乱疫情相关菌株为genotype A型菌,而绝大多数环境分

  17. Development of fowl cholera vaccine: I. Protection of Pasteurella multocida local isolate vaccine against challenge of homologous and heterologous strains.

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    Supar

    2001-03-01

    Full Text Available Pasteurella multocida locally isolated from chicken and ducks (BCC 299, BCC 2331, DY1, DY2, 12TG, 15TG andimported strains (BCC 1359, 1362; HEDDLESTON group 1 and 6 respectively had been tested for its pathogenicity in theprevious study. The aims of this experiment were to study the preparation of local isolate pasteurellosis vaccines and to determine the protective effect of that vaccines in chicken against the highly pathogenic local isolates of P. multocida. Killed monovalent, bivalent and polyvalent pasteurellosis vaccines were prepared and each was adjunvanted with aluminum hydroxide gel at a final concentration of 1.5% and the cell concentration was equal to the No 10 of MacFarland tube standard. Each of the vaccine prepared was used to vaccinated on a group of six week old of layer chicken (8 per group. Each chicken was subcutaneously injected with 0.2 ml of vaccine, four weeks later each was boostered with similar vaccine with the same dose. Two weeks after giving the boostered vaccine each group of chicken were challenged, half with life bacterium of P. Multocida BCC 2331 and other with DY2. Any chick which survive after challenge was designated as protected by vaccination. Before vaccination 1 ml of blood was drawn from each of chicken and then two weeks apart up to challenge. Serum from each sample was separated and kept in deep freeze until tested by enzyme-linked immunosorbent assay (ELISA. Chicken vaccinated with killed whole cell P. multocida vaccines of monovalent (BCC 2331 or DY2 and bivalent (BCC 2331 + DY2 were protected against challenge with live bacterium of either BCC 2331 or DY2 at rate 67-100%. There was no protection in chicken vaccinated with either BCC 299, DY1, 12TG, 15TG, BCC 1359, or 1362 killed vaccine. Similarly no protection of chicken vaccinated with either DY1 + BCC299, 12TG + 15TG or BCC 1359 + BCC 1362 bivalent vaccines. The protection rate of the polyvalent local isolate vaccine was at average 50-75%. All

  18. Cholera toxin expression by El Tor Vibrio cholerae in shallow culture growth conditions.

    Science.gov (United States)

    Cobaxin, Mayra; Martínez, Haydee; Ayala, Guadalupe; Holmgren, Jan; Sjöling, Asa; Sánchez, Joaquín

    2014-01-01

    Vibrio cholerae O1 classical, El Tor and O139 are the primary biotypes that cause epidemic cholera, and they also express cholera toxin (CT). Although classical V. cholerae produces CT in various settings, the El Tor and O139 strains require specific growth conditions for CT induction, such as the so-called AKI conditions, which consist of growth in static conditions followed by growth under aerobic shaking conditions. However, our group has demonstrated that CT production may also take place in shallow static cultures. How these type of cultures induce CT production has been unclear, but we now report that in shallow culture growth conditions, there is virtual depletion of dissolved oxygen after 2.5 h of growth. Concurrently, during the first three to 4 h, endogenous CO2 accumulates in the media and the pH decreases. These findings may explain CT expression at the molecular level because CT production relies on a regulatory cascade, in which the key regulator AphB may be activated by anaerobiosis and by low pH. AphB activation stimulates TcpP synthesis, which induces ToxT production, and ToxT directly stimulates ctxAB expression, which encodes CT. Importantly, ToxT activity is enhanced by bicarbonate. Therefore, we suggest that in shallow cultures, AphB is activated by initial decreases in oxygen and pH, and subsequently, ToxT is activated by intracellular bicarbonate that has been generated from endogenous CO2. This working model would explain CT production in shallow cultures and, possibly, also in other growth conditions.

  19. The hows and whys of constructing a native recombinant cholera vaccine

    Science.gov (United States)

    Boustanshenas, Mina; Bakhshi, Bita

    2014-01-01

    Emergence of different ctxB genotypes within virulent Vibrio cholerae populations accentuates the need to develop a vaccine that has the potential to protect against all cholera toxin genotypes. Oral administration of rCTB—alone and in combination with 2 dominant domestic killed whole cells of V. cholerae (O1 Ogawa El Tor and O1 Inaba El Tor) plus one standard V. cholerae (O1 Ogawa classic ATCC 14035)—has shown satisfactory protection as a potent vaccine candidate against toxigenic V. cholerae. PMID:24165439

  20. Methods to assess the impact of mass oral cholera vaccination campaigns under real field conditions.

    Directory of Open Access Journals (Sweden)

    Jacqueline Deen

    Full Text Available There is increasing interest to use oral cholera vaccination as an additional strategy to water and sanitation interventions against endemic and epidemic cholera. There are two internationally-available and WHO-prequalified oral cholera vaccines: an inactivated vaccine containing killed whole-cells of V. cholerae O1 with recombinant cholera toxin B-subunit (WC/rBS and a bivalent inactivated vaccine containing killed whole cells of V. cholerae O1 and V. cholerae O139 (BivWC. The efficacy, effectiveness, direct and indirect (herd protection conferred by WC/rBS and BivWC are well established. Yet governments may need local evidence of vaccine impact to justify and scale-up mass oral cholera vaccination campaigns. We discuss various approaches to assess oral cholera vaccine protection, which may be useful to policymakers and public health workers considering deployment and evaluation of the vaccine.

  1. Serotype cycles in cholera dynamics

    OpenAIRE

    Koelle, Katia; Pascual, Mercedes; Yunus, Md.

    2006-01-01

    Interest in understanding strain diversity and its impact on disease dynamics has grown over the past decade. Theoretical disease models of several co-circulating strains indicate that incomplete cross-immunity generates conditions for strain-cycling behaviour at the population level. However, there have been no quantitative analyses of disease time-series that are clear examples of theoretically expected strain cycling. Here, we analyse a 40-year (1966–2005) cholera time-series from Banglade...

  2. Detection of Vibrio cholerae and Acanthamoeba species from same natural water samples collected from different cholera endemic areas in Sudan

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    Saeed Amir

    2011-04-01

    Full Text Available Abstract Background Vibrio cholerae O1 and V. cholerae O139 infect humans, causing the diarrheal and waterborne disease cholera, which is a worldwide health problem. V. cholerae and the free-living amoebae Acanthamoeba species are present in aquatic environments, including drinking water and it has shown that Acanthamoebae support bacterial growth and survival. Recently it has shown that Acanthamoeba species enhanced growth and survival of V. cholerae O1 and O139. Water samples from different cholera endemic areas in Sudan were collected with the aim to detect both V. cholerae and Acanthamoeba species from same natural water samples by polymerase chain reaction (PCR. Findings For the first time both V. cholerae and Acanthamoeba species were detected in same natural water samples collected from different cholera endemic areas in Sudan. 89% of detected V. cholerae was found with Acanthamoeba in same water samples. Conclusions The current findings disclose Acanthamoedae as a biological factor enhancing survival of V. cholerae in nature.

  3. Application of a paper based device containing a new culture medium to detect Vibrio cholerae in water samples collected in Haiti.

    Science.gov (United States)

    Briquaire, Romain; Colwell, Rita R; Boncy, Jacques; Rossignol, Emmanuel; Dardy, Aline; Pandini, Isabelle; Villeval, François; Machuron, Jean-Louis; Huq, Anwar; Rashed, Shah; Vandevelde, Thierry; Rozand, Christine

    2017-02-01

    Cholera is now considered to be endemic in Haiti, often with increased incidence during rainy seasons. The challenge of cholera surveillance is exacerbated by the cost of sample collection and laboratory analysis. A diagnostic tool is needed that is low cost, easy-to-use, and able to detect and quantify Vibrio cholerae accurately in water samples within 18-24h, and perform reliably in remote settings lacking laboratory infrastructure and skilled staff. The two main objectives of this study were to develop and evaluate a new culture medium embedded in a new diagnostic tool (PAD for paper based analytical device) for detecting Vibrio cholerae from water samples collected in Haiti. The intent is to provide guidance for corrective action, such as chlorination, for water positive for V. cholerae epidemic strains. For detecting Vibrio cholerae, a new chromogenic medium was designed and evaluated as an alternative to thiosulfate citrate bile salts sucrose (TCBS) agar for testing raw water samples. Sensitivity and specificity of the medium were assessed using both raw and spiked water samples. The Vibrio cholerae chromogenic medium was proved to be highly selective against most of the cultivable bacteria in the water samples, without loss of sensitivity in detection of V. cholerae. Thus, reliability of this new culture medium for detection of V. cholerae in the presence of other Vibrio species in water samples offers a significant advantage. A new paper based device containing the new chromogenic medium previously evaluated was compared with reference methods for detecting V. cholerae from spiked water sample. The microbiological PAD specifications were evaluated in Haiti. More precisely, a total of 185 water samples were collected at five sites in Haiti, June 2014 and again in June 2015. With this new tool, three V. cholerae O1 and 17 V. cholerae non-O1/O139 strains were isolated. The presence of virulence-associated and regulatory genes, including ctxA, zot, ace, and tox

  4. Cholera Treatment

    Science.gov (United States)

    ... Public Health in Haiti Haiti Pre-decision Brief Cholera in Haiti: One Year Later Related Links Healthy Water Global Water, Sanitation, & Hygiene (WASH) The Safe Water System Division of Foodborne, Waterborne, and Environmental Diseases Get Email Updates To receive email updates about ...

  5. 2005年-2010年武汉市霍乱弧菌监测分析%Analysis of surveillance on Vibrio Cholerae between 2005 and 2010 in Wuhan city

    Institute of Scientific and Technical Information of China (English)

    熊燕; 江元山; 陈智; 王芳; 龙一兵; 周军波

    2012-01-01

    目的:对2005年-2010年霍乱弧菌监测中分离的25株疑似菌株进行系统鉴定.方法:分离培养、血清学试验、系统生化鉴定( VITEK32)、PCR检测其分型及毒素基因、药敏试验.结果:25株菌鉴定为霍乱弧菌的有21株,与霍乱弧菌发生交叉凝集4株;病例监测主要为0139霍乱弧菌、外环境监测主要为01群霍乱弧菌;病例监测菌株携带霍乱弧菌毒素基因(ctxAB)、其他菌株均不携带霍乱弧菌毒素基因(ctxAB).药敏实验结果对诺氟沙星和环丙沙星敏感,对其他抗生素有不同程度的耐药.结论:武汉市霍乱弧菌在病例监测中主要为0139群霍乱弧菌产毒株、外环境监测中主要为O1群霍乱弧菌非产毒株,不同型别菌株存在不同程度耐药性.%Objective: 25 suspected strains of Vibrio cholerae isolated between 2005 and 2010 in the Vibrio cholerae surveillance program were identified and analyzed by different kinds of assays. Methods; The isolates were analyzed by conventional diagnostic methods and PCR assays for their molecular typing and virulence-related genes, and drug sensitivity test. Results: In 25 suspected strains of Vibrio cholerae, there were 21 vibrio cholerae isolates and four strains cross -agglutinating with Vibrio cholerae,respectively. Vibrio Cholerae Serogroup 0139 came mostly from patients,and Vibrio cholerae Serogroup 01 was mostly isolated from natural circumstances. Six strains of Vibrio cholerae Serogroup 0139 were positive for ctxAB gene,and others were negative. The drug sensitivity tests indicated that all suspected strains of Vibrio cholerae were sensitive to norfloxacin and ciprofloxacin, and exhibited varying degrees of resistance to other antibiotics. Conclusion: The suspected strains of Vibrio cholerae isolated from patients were mostly Vibrio Cholerae Serogroup 0139, and Vibrio cholerae Serogroup 01 came mostly from natural circumstances. All suspected isolates showed varying degrees of resistance to

  6. epidemiological and Clinical Characteristics of 28 cases of Cholera

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    The data of 35 246 patients with intestinal diseases were retrospectively analyzed, 28 cases of cholera patients were screened in 17 years, of which 23 cases had suspicious unclean food history, 10 cases were migrant workers, 8 cases had history of coastal city tour in one week. All of the 28 patients were positive for Vibrio cholerae culture, 19 cases were identiifed as O1 serotype Ogawa and 6 were identiifed as O1 serotype Inaba, 3 were identified as O139. Twenty-three patients were mild, five cases were moderate, patients with severe diseases were not found. It was found in this study that O1 serotype Vibrio cholerae was still dominant, 82%of cholera patients were mild cases. Tourists who had a incompletely heated seafood intake history and migrant people are susceptible to cholera.

  7. Chimeric adaptor proteins translocate diverse type VI secretion system effectors in Vibrio cholerae.

    Science.gov (United States)

    Unterweger, Daniel; Kostiuk, Benjamin; Ötjengerdes, Rina; Wilton, Ashley; Diaz-Satizabal, Laura; Pukatzki, Stefan

    2015-08-13

    Vibrio cholerae is a diverse species of Gram-negative bacteria, commonly found in the aquatic environment and the causative agent of the potentially deadly disease cholera. These bacteria employ a type VI secretion system (T6SS) when they encounter prokaryotic and eukaryotic competitors. This contractile puncturing device translocates a set of effector proteins into neighboring cells. Translocated effectors are toxic unless the targeted cell produces immunity proteins that bind and deactivate incoming effectors. Comparison of multiple V. cholerae strains indicates that effectors are encoded in T6SS effector modules on mobile genetic elements. We identified a diverse group of chimeric T6SS adaptor proteins required for the translocation of diverse effectors encoded in modules. An example for a T6SS effector that requires T6SS adaptor protein 1 (Tap-1) is TseL found in pandemic V. cholerae O1 serogroup strains and other clinical isolates. We propose a model in which Tap-1 is required for loading TseL onto the secretion apparatus. After T6SS-mediated TseL export is completed, Tap-1 is retained in the bacterial cell to load other T6SS machines.

  8. Pathogenicity of Vibrio anguillarum serogroup O1 strains compared to plasmids, outer membrane protein profiles and siderophore production

    DEFF Research Database (Denmark)

    Pedersen, K.; Gram, Lone; Austin, D.A.;

    1997-01-01

    The virulence of 18 strains of Vibrio anguillarum serogroup 01 was compared to plasmid content, expression of siderophores and outer membrane proteins. All strains, irrespective of plasmid content, produced siderophores and inducible outer membrane proteins under iron-limited conditions. Only...... strains that carried the 67 kbp virulence plasmid or derivatives of it produced the outer membrane protein, OM2. All virulent strains harboured the 67 kbp plasmid or derivatives of it, indicating its importance for virulence. However, some strains carrying the virulence plasmid or a derivative of it...

  9. Zebrafish as a natural host model for Vibrio cholerae colonization and transmission.

    Science.gov (United States)

    Runft, Donna L; Mitchell, Kristie C; Abuaita, Basel H; Allen, Jonathan P; Bajer, Sarah; Ginsburg, Kevin; Neely, Melody N; Withey, Jeffrey H

    2014-03-01

    The human diarrheal disease cholera is caused by the aquatic bacterium Vibrio cholerae. V. cholerae in the environment is associated with several varieties of aquatic life, including insect egg masses, shellfish, and vertebrate fish. Here we describe a novel animal model for V. cholerae, the zebrafish. Pandemic V. cholerae strains specifically colonize the zebrafish intestinal tract after exposure in water with no manipulation of the animal required. Colonization occurs in close contact with the intestinal epithelium and mimics colonization observed in mammals. Zebrafish that are colonized by V. cholerae transmit the bacteria to naive fish, which then become colonized. Striking differences in colonization between V. cholerae classical and El Tor biotypes were apparent. The zebrafish natural habitat in Asia heavily overlaps areas where cholera is endemic, suggesting that zebrafish and V. cholerae evolved in close contact with each other. Thus, the zebrafish provides a natural host model for the study of V. cholerae colonization, transmission, and environmental survival.

  10. Surface expression of Helicobacter pylori HpaA adhesion antigen on Vibrio cholerae, enhanced by co-expressed enterotoxigenic Escherichia coli fimbrial antigens.

    Science.gov (United States)

    Tobias, Joshua; Lebens, Michael; Wai, Sun Nyunt; Holmgren, Jan; Svennerholm, Ann-Mari

    2017-02-17

    Helicobacter pylori infection can cause peptic ulceration and is associated with gastric adenocarcinoma. This study aimed to construct and characterize a non-virulent Vibrio cholerae O1 strain, which grows more rapidly than H. pylori, as vector for H. pylori antigens for possible use as a vaccine strain against H. pylori. This was done by recombinant expression of the H. pylori adhesion antigen HpaA alone or, as a proof of principle, together with different colonization factor (CF) antigens of enterotoxigenic Escherichia coli (ETEC) which may enhance immune responses against HpaA. A recombinant V. cholerae strain co-expressing HpaA and a fimbrial CF antigens CFA/I or CS5, but not the non-fimbrial CF protein CS6, was shown to express larger amounts of HpaA on the surface when compared with the same V. cholerae strain expressing HpaA alone. Mutations in the CFA/I operon showed that the chaperon, possibly together with the usher, was involved in enhancing the surface expression of HpaA. Oral immunization of mice with formaldehyde-inactivated recombinant V. cholerae expressing HpaA alone or together with CFA/I induced significantly higher serum antibody responses against HpaA than mice similarly immunized with inactivated HpaA-expressing H. pylori bacteria. Our results demonstrate that a non-virulent V. cholerae strain can be engineered to allow strong surface expression of HpaA, and that the expression can be further increased by co-expressing it with ETEC fimbrial antigens. Such recombinant V. cholerae strains expressing HpaA, and possibly also other H. pylori antigens, may have the potential as oral inactivated vaccine candidates against H. pylori.

  11. What is cholera?

    DEFF Research Database (Denmark)

    Tamason, Charlotte Crim; Tulsiani, Suhella; Siddique, A.;

    2016-01-01

    a third ofthe respondents did not associate diarrhea with cholera or mentioned symptoms that could not be caused by cholera (29%). Approximately half of the respondents associated water with the cause of cholera (56%) and only 8% associated cholera with sanitation or hygiene. Shame and stigma (54%) were...

  12. A study on the pathogenic characteristics and traceability of Vibrio cholera strains circulated in Hubei province in 2012%2012年湖北省霍乱的病原学研究及溯源分析

    Institute of Scientific and Technical Information of China (English)

    张婷; 杨红梅; 程均福; 吕静; 刘公平; 李国明

    2013-01-01

      目的分析2012年湖北省霍乱流行的病原学特征,应用脉冲场凝胶电泳分析各疫情菌株之间的遗传相关性,查找霍乱传染来源,为制定防治措施提供依据。方法对35株霍乱弧菌菌株进行常规生化鉴定和毒素基因检验,以及药敏试验,采用脉冲场凝胶电泳( PFGE)技术获得电泳酶切指纹图谱并对图谱进行聚类分析。结果35株霍乱弧菌经检验均为霍乱 O139群,产毒株占71.42%,霍乱弧菌耐药结果显示四环素、复方新诺明、利福平耐药率分别为57.14%、88.57%、80.00%。 PFGE电泳图谱条带总相似率为80%~100%,具有一定的同源性。三起疫情中相同地区的大部分菌株都聚为一类,同源性为100%,提示为相同菌株感染所致;仅来源于荆州地区甲鱼中分离的菌株单独为一类与其他菌株不同。结论2012年湖北省霍乱弧菌的优势菌株为O139群,大部分产毒。药敏结果显示,菌株对四环素、复方新诺明、利福平大部分耐药;对亚胺培南、头孢曲松高度敏感;疫情中相同地区的大部分菌株聚为一类属于同一来源,不同地区的菌株有一定的变异,几起疫情暴发均与聚餐有关,所以注意食物卫生,从甲鱼中分离的菌株不产毒,提示甲鱼可能并不是疫情的主要毒株来源,应密切关注海、水产品的监测情况。%Objective To investigate the pathogenic characteristics of Vibrio cholera strains isola-ted from Hubei province in 2012 , and to identify the source of infection by analyzing their genetic correla-tions.Methods The biochemical identification , toxin gene detection and drug susceptibility test were car-ried out to analyze a total of 35 Vibrio cholera strains isolated from three epidemic areas .Comparison of ge-nomic DNA fingerprints and cluster analysis among isolates of Vibrio cholera was conducted by using pulsed-field gel electrophoresis ( PFGE

  13. Protsome analysis of the total protein in Vibrio cholerae by 2-DE%霍乱弧菌O1群有毒株和无毒株的蛋白质谱特征分析

    Institute of Scientific and Technical Information of China (English)

    陈守义; 刘俊华; 张欣强; 胡玉山; 李钏华; 王鸣

    2008-01-01

    Objective To study the different proteins of human and environment Vibrio cholerae (V. cholerae) by two dimensional electrophoresis (2-DE) and TOF-TOF-MS. Methods Total cellular pro-tein was extracted by lysate from V. cholerae, and proteins were separated by 2-DE under immobilized pH gradients(IPG), then electrophoregrams were dealed with coomassie brilliant blue, and analyzed by Im-ageMaster 2D Elite 5.0, finally the different protein spots were identified by TOF-TOF-MS. Results High repetitive 2-DE maps were obtained. 2-DE and image analysis revealed 1032±22 protein spots, and PI value was among 4.00 to 7.20. Matching spots were 1025±24 in two repeats electrophoregrams and matching ratio was 96.30%. Conclusion The different protein spots were successfully established with high quality and sharpness separation by 2-DE and TOF-TOF-MS, which stands as a valuable resource for proteomics re-search of V. cholerae.%目的 利用双向电泳和质谱鉴定技术探讨环境和人体中霍乱弧菌蛋白表达的差异.方法 利用适当的裂解液处理霍乱弧菌,提取全菌蛋白;采用pH梯度等电聚焦对全菌蛋白进行双向电泳;考马斯亮蓝染色后获得双向电泳图谱,并利用ImageMaster 2D Elite 5.0图像分析软件进行分析,所得的数据采用SPSS15.0进行统计分析,找出差异蛋白;在此基础上,胰蛋白酶消化这些特殊差异蛋白,并进行质谱(MALDI-TOF-MS)分析.结果 获得1032±22个蛋白斑点,蛋白主要集中在等电点(PI)4.00~7.20之间,重复胶的匹配点数为1025±24,匹配率为96.30%;发现了有明显差异的21个蛋白点,并用质谱鉴定了其中4种蛋白.结论 获得了环境和人体中霍乱弧菌蛋白的差异表达蛋白.

  14. Suppression of Virulence of Toxigenic Vibrio cholerae by Anethole through the Cyclic AMP (cAMP)-cAMP Receptor Protein Signaling System.

    Science.gov (United States)

    Zahid, M Shamim Hasan; Awasthi, Sharda Prasad; Asakura, Masahiro; Chatterjee, Shruti; Hinenoya, Atsushi; Faruque, Shah M; Yamasaki, Shinji

    2015-01-01

    Use of natural compounds as antivirulence drugs could be an alternative therapeutic approach to modify the outcome of bacterial infections, particularly in view of growing resistance to available antimicrobials. Here, we show that sub-bactericidal concentration of anethole, a component of sweet fennel seed, could suppress virulence potential in O1 El Tor biotype strains of toxigenic Vibrio cholerae, the causative agent of the ongoing 7th cholera pandemic. The expression of cholera toxin (CT) and toxin coregulated pilus (TCP), the major virulence factors of V. cholerae, is controlled through a regulatory cascade involving activation of ToxT with synergistic coupling interaction of ToxR/ToxS with TcpP/TcpH. We present evidence that anethole inhibits in vitro expression of CT and TCP in a toxT-dependent but toxR/toxS-independent manner and through repression of tcpP/tcpH, by using bead-ELISA, western blotting and quantitative real-time RT-PCR assays. The cyclic AMP (cAMP)-cAMP receptor protein (CRP) is a well-studied global signaling system in bacterial pathogens, and this complex is known to suppress expression of tcpP/tcpH in V. cholerae. We find that anethole influences the virulence regulatory cascade by over-expressing cyaA and crp genes. Moreover, suppression of toxigenic V. cholerae-mediated fluid accumulation in ligated ileum of rabbit by anethole demonstrates its potentiality as an antivirulence drug candidate against the diseases caused by toxigenic V. cholerae. Taken altogether, these results revealing a mechanism of virulence inhibition in V. cholerae by the natural compound anethole, may have relevance in designing antivirulence compounds, particularly against multiple antibiotic resistant bacterial pathogens.

  15. Household Transmission of Vibrio cholerae in Bangladesh.

    Directory of Open Access Journals (Sweden)

    Jonathan D Sugimoto

    2014-11-01

    Full Text Available Vibrio cholerae infections cluster in households. This study's objective was to quantify the relative contribution of direct, within-household exposure (for example, via contamination of household food, water, or surfaces to endemic cholera transmission. Quantifying the relative contribution of direct exposure is important for planning effective prevention and control measures.Symptom histories and multiple blood and fecal specimens were prospectively collected from household members of hospital-ascertained cholera cases in Bangladesh from 2001-2006. We estimated the probabilities of cholera transmission through 1 direct exposure within the household and 2 contact with community-based sources of infection. The natural history of cholera infection and covariate effects on transmission were considered. Significant direct transmission (p-value<0.0001 occurred among 1414 members of 364 households. Fecal shedding of O1 El Tor Ogawa was associated with a 4.9% (95% confidence interval: 0.9%-22.8% risk of infection among household contacts through direct exposure during an 11-day infectious period (mean length. The estimated 11-day risk of O1 El Tor Ogawa infection through exposure to community-based sources was 2.5% (0.8%-8.0%. The corresponding estimated risks for O1 El Tor Inaba and O139 infection were 3.7% (0.7%-16.6% and 8.2% (2.1%-27.1% through direct exposure, and 3.4% (1.7%-6.7% and 2.0% (0.5%-7.3% through community-based exposure. Children under 5 years-old were at elevated risk of infection. Limitations of the study may have led to an underestimation of the true risk of cholera infection. For instance, available covariate data may have incompletely characterized levels of pre-existing immunity to cholera infection. Transmission via direct exposure occurring outside of the household was not considered.Direct exposure contributes substantially to endemic transmission of symptomatic cholera in an urban setting. We provide the first estimate of

  16. Invasive Vibrio cholerae Infection Following Burn Injury

    Science.gov (United States)

    2008-06-01

    as asymptomatic col- onization, otitis, gastroenteritis , soft-tissue infection, sepsis, or even cerebritis. In contrast, epidemic V. cholerae (O-1 or...review of the available literature is presented in Table 1. CONCLUSION Infection with invasive Vibrio species bacteria (e.g. Vibrio vulnificus

  17. Identification of two vicinal operons for the degradation of 2-aminobenzenesulfonate encoded on plasmid pSAH in Alcaligenes sp. strain O-1.

    Science.gov (United States)

    Ruff, Jürgen; Smits, Theo H M; Cook, Alasdair M; Schleheck, David

    2010-05-30

    Alcaligenes sp. strain O-1 inducibly deaminates 2-aminobenzenesulfonate (ABS) via dioxygenation to 3-sulfocatechol, which is desulfonated during meta ring-cleavage to yield 2-hydroxymuconate. This intermediate is transformed through the oxalocrotonate-branch of the sulfocatechol meta-pathway (Scm). The complete pathway is encoded on the 180-kb plasmid pSAH, 20kb of which was sequenced. Twenty open reading frames (ORFs) were detected. Two clusters (abs and scm) with degradative genes were surrounded by several transposon-related ORFs. The six genes of the abs cluster were shown to be co-transcribed, and contained the genes for two characterised subunits of the oxygenase component of the ABS-dioxygenase system, and genes putatively encoding ABS-transport functions with similarities to (a) an ABC-type transporter system and (b) a putative major facilitator superfamily transporter. No gene encoding the reductase for the oxygenase system was present in the abs gene cluster, but a candidate gene was found in the scm cluster. The seven-gene scm cluster was also transcribed as single polycistronic message. Functions could be attributed to the gene products, but one enzyme, which was shown to be present, 2-hydroxymuconate isomerase, was not encoded in the scm cluster. No transcriptional regulator was found. This genetic information on the degradation of ABS in strain O-1 provides another example of both split operons and dispersed pathway genes.

  18. Twin outbreak of cholera in rural North Karnataka, India

    Directory of Open Access Journals (Sweden)

    Shuchismita Dey

    2014-01-01

    Full Text Available Background & objectives: Successive outbreaks of acute watery diarrhoea occurred in Talikoti and Harnal, located in Bijapur District of the southern Indian s0 tate of Karnataka, in July and August 2012, respectively. These outbreaks were investigated to identify the aetiology and epidemiology. Methods: Information was collected from the local population and health centres. Stool and water samples were collected from the admitted patients and their drinking water sources. Standard microbiological and PCR techniques were employed for isolation and characterization of the pathogen. Results: While 101 people (0.38% were affected in Talikoti, 200 (20.94% were affected in Harnal which is a small remote village. All age groups were affected but no death occurred. While the outbreak was smaller, longer and apparently spread by person to person contact in Talikoti, it occurred as a single source flash outbreak at Harnal. A single clone of toxigenic Vibrio cholerae O1 Ogawa biotype El Tor was isolated from the two stool samples obtained from Talikoti and subsequently from three of five stool samples obtained from Harnal indicating village to village spread of the aetiological agent. Striking similarity in antibiotic resistance profiles of these isolates with a particular strain isolated from the city of Belgaum, 250 km away, in 2010, prompted tracking the lineage of the V. cholerae isolates by DNA fingerprinting. Random amplified polymorphic DNA (RAPD fingerprinting assay helped confirm the origin of the incriminating strain to Belgaum. Interpretation & conclusions: Our study reported the first twin outbreak of cholera in two remote areas of Bijapur district, Karnataka, south India. It also indicated the need for immediate preparedness to deal with such emergencies.

  19. Outbreak-associated Vibrio cholerae genotypes with identical pulsotypes, Malaysia, 2009.

    Science.gov (United States)

    Teh, Cindy Shuan Ju; Suhaili, Zarizal; Lim, King Ting; Khamaruddin, Muhamad Afif; Yahya, Fariha; Sajili, Mohd Hailmi; Yeo, Chew Chieng; Thong, Kwai Lin

    2012-07-01

    A cholera outbreak in Terengganu, Malaysia, in November 2009 was caused by 2 El Tor Vibrio cholerae variants resistant to typical antimicrobial drugs. Evidence of replacement of treatable V. cholerae infection in the region with antimicrobial-resistant strains calls for increased surveillance and prevention measures.

  20. Epidemiology, determinants and dynamics of cholera in Pakistan: gaps and prospects for future research.

    Science.gov (United States)

    Naseer, Maliha; Jamali, Tanzil

    2014-11-01

    Cholera is one of the notifiable endemic diseases in Pakistan, but the reporting of cholera cases is still unsatisfactory. Most of the diagnosed cases are never reported to the relevant authorities. In the year 1993 - 2005, the country did not report any single case of cholera to the WHO. The objectives of this review were to understand the epidemiology and to identify the possible determinants of cholera infection in Pakistan. Medscape, Medline, PakMedinet and PubMed, was searched, using key words, epidemiology and determinants of cholera infection in Pakistan during 1995 - 2010. Morbidity and mortality due to cholera infection during 1995 - 2010, without any language restriction. Out of 27 articles published between 1995 - 2010, 17 articles were included in the review. Vibrio cholerae O139 identified as a major cause of infection in older age group, while O1 biotype of cholera as a predominant cause of cholera among young individuals. Mainly reported determinants of cholera in Pakistan include poor sanitation and hygiene practices, increased population density in urban areas, leading to rapid and unplanned urbanization of the major cities and climate change due to increased environmental pollution in Pakistan are plausible factors for endemicity of cholera in Pakistan. Cholera reporting as a notifiable disease to the relevant departments and timely action can prevent the risk of outbreaks. There is a need to identify specific behavioral and environmental determinants responsible for outbreaks and epidemics of cholera in Pakistan which can help to design appropriate preventive and control interventions.

  1. 嘉兴市霍乱弧菌分离株检测分析%Analysis of Vibrio cholerae isolates in Jiaxing

    Institute of Scientific and Technical Information of China (English)

    何培彦; 高雯洁; 葛志坚; 王恒辉; 燕勇

    2013-01-01

    目的 比较分析嘉兴市霍乱弧菌菌株的血清型、毒力基因和分子分型特征.方法 采用血清学和分子生物学方法对近年来分离到13株霍乱弧菌菌株进行血清型分布、毒力基因(ctxA、ace、zot、tcpA、cri和rtxA)携带和ERIC PCR分型研究.结果 13株霍乱弧菌菌株,2株为O139群霍乱弧菌,11株为O1群霍乱弧菌,其中小川型9株,稻叶型2株.2株O139群霍乱弧菌菌株携带3种毒力基因;11株O1群霍乱弧菌菌株,除1株携带4种毒力基因外,其余10株携带全部6种毒力基因.ERIC-PCR分型分析显示,不同的霍乱弧菌菌株之间ERIC-PCR型别表现出一定的遗传多样性.结论 嘉兴市霍乱弧菌菌株大多携带多种毒力基因,而且来源多样,防控工作任重而道远.%Objective To analyze the serotype,virulence genes and molecular typing of Vibrio cholerae strains isolated in Jiaxing.Methods The serotype,virulence genes (ctxA,ace,zot,tcpA,cri and rtxA) and ERIC-PCR typing of 13 Vibrio cholerae strains isolated in recent years were studied by serological and molecular biological methods.Results Out of 13 Vibrio cholerae strains,2 were identified as serogroup O139 and 11 were serogroup O1 which included 9 Ogawa and 2 Inaba strains.2 O139 strains carried 3 virulence genes.10 O1 strains harbored all 6 virulence genes and only one carried 4.ERIC-PCR typing analysis showed that the genetic diversity existed among different Vibrio cholerae strains.Conclusions Majority of Vibrio cholerae strains isolated in Jiaxing carried multiple virulence genes with various origins.Therefore,more work needs to be done for the prevention.

  2. Cholera Prevention and Control

    Science.gov (United States)

    ... name="commit" type="submit" value="Submit" /> Prevention & Control Recommend on Facebook Tweet Share Compartir Prevention of ... of cholera and other diarrheal disease prevention. Prevention Control Topics Six Basic Cholera Prevention Messages I nfection ...

  3. What is cholera?

    DEFF Research Database (Denmark)

    Tamason, Charlotte Crim; Tulsiani, Suhella; Siddique, A.;

    2016-01-01

    Background: Cholera has afflicted the Indian sub-continent for centuries, predominantly in West Bengal and modern-day Bangladesh. This preliminary study aims to understand the current level of knowledge of cholera in female Bangladeshi caretakers, which is important in the outcome of the disease...... and its spread. A pilot study was conducted among 85 women in Bangladesh using qualitative questionnaires to explore the ability of female caretakers in identifying cholera and its transmission. Findings: The survey revealed that though all the female caretakers were aware of the term “cholera,” nearly...... a third ofthe respondents did not associate diarrhea with cholera or mentioned symptoms that could not be caused by cholera (29%). Approximately half of the respondents associated water with the cause of cholera (56%) and only 8% associated cholera with sanitation or hygiene. Shame and stigma (54%) were...

  4. Multi-locus variable number tandem repeat analysis of Vibrio cholerae isolates from 2012 to 2013 cholera outbreaks in Iran.

    Science.gov (United States)

    Ranjbar, R; Sadeghy, J; Shokri Moghadam, M; Bakhshi, B

    2016-08-01

    Cholera remains to be an international threat, with high rates of illness and death. In 2012 and 2013, two cholera outbreak happened in Iran, affecting lots of people. Vibrio cholerae O1 was confirmed as the etiological agent. Source identification and controlling the spread of the cholera disease are two critical approaches in cholera outbreaks. In this study, thirty V. cholerae O1 isolates were selected and has been evaluated for antimicrobial resistant as well as molecular typing by multilocus variable-number tandem-repeat analysis (MLVA) method. Twenty-nine (97%) isolates were sero-grouped as El Tor (one isolate was classical) and 100% were related to Inaba serotype. All of the isolates were susceptible to ciprofloxacin, chloramphenicol, ampicillin and gentamicin. On the other hand, 60% of the isolates were MDR (resistant to 3 or more classes). There were three resistance patterns. The most prevalent pattern was resistance to streptomycin, erythromycin, trimethoprim-sulfamethoxazole, and tetracycline (ST-SXT-E-T) which was seen in 50% of isolates. Using MLVA method 14 MLVA types were identified. MLVA type 2 (5-7-7-16-15) accounted for 43% of isolates. Isolates with the same genotype often did not have the same antibiogram. Overall, the data indicate that the Iranian V. cholerae were MDR and clonaly related. Furthermore, the results of this study shows that MLVA can be used as useful method for V. cholerae genotyping in epidemiological investigations.

  5. Low detection of Vibrio cholerae carriage in healthcare workers returning to 12 Latin American countries from Haiti.

    Science.gov (United States)

    Llanes, R; Somarriba, L; Hernández, G; Bardaji, Y; Aguila, A; Mazumder, R N

    2015-04-01

    SUMMARY This investigation was undertaken to characterize the prevalence of intestinal Vibrio cholerae in healthcare workers (HCWs) returning from Haiti due to the ongoing cholera epidemic. Eight hundred and fifty asymptomatic HCWs of the Cuban Medical Brigade, who planned to leave Haiti, were studied by laboratory screening of stool culture for V. cholerae. A very low percentage (0.23%) of toxigenic V. cholerae serogroup O1, serotype Ogawa was found. To the best of our knowledge, this study represents the largest reported screening study for V. cholerae infection in asymptomatic HCWs returning from a cholera-affected country. Cholera transmission to health personnel highlights a possible risk of transmitting cholera during mobilization of the population for emergency response. Aid workers are encouraged to take precautions to reduce their risk for acquiring cholera and special care should be taken by consuming safe water and food and practising regular hand washing.

  6. Cholera in Zimbabwe

    NARCIS (Netherlands)

    Pruyt, E.

    2009-01-01

    By the end of December 2008, alarming reports and articles concerning the cholera outbreak in Zimbabwe received plenty of international media coverage. By that time nearly 30000 cases of cholera infections and 1600 cholera deaths had been reported. In the first week of January 2009, a System Dynamic

  7. Characterisation of cholera toxin by liquid chromatography - Electrospray mass spectrometry

    NARCIS (Netherlands)

    Baar, B.L.M. van; Hulst, A.G.; Wils, E.R.J.

    1999-01-01

    Cholera toxin, one of the toxins that may be generated by various strains of the bacterium Vibrio cholerae, can be considered as a substance possibly used in biological warfare. The possibilities of characterising the toxin by liquid chromatography electrospray mass spectrometry (LC-ES-MS) were inve

  8. Malonate inhibits virulence gene expression in Vibrio cholerae.

    Science.gov (United States)

    Minato, Yusuke; Fassio, Sara R; Häse, Claudia C

    2013-01-01

    We previously found that inhibition of the TCA cycle, either through mutations or chemical inhibition, increased toxT transcription in Vibrio cholerae. In this study, we found that the addition of malonate, an inhibitor of succinate dehydrogenase (SDH), decreased toxT transcription in V. cholerae, an observation inconsistent with the previous pattern observed. Unlike another SDH inhibitor, 2-thenoyltrifluoroacetone (TTFA), which increased toxT transcription and slightly inhibited V. cholerae growth, malonate inhibited toxT transcription in both the wild-type strain and TCA cycle mutants, suggesting malonate-mediated inhibition of virulence gene expression is independent to TCA cycle activity. Addition of malonate also inhibited ctxB and tcpA expressions but did not affect aphA, aphB, tcpP and toxR expressions. Malonate inhibited cholera toxin (CT) production in both V. cholerae classical biotype strains O395N1 and CA401, and El Tor biotype strain, N16961. Consistent with previous reports, we confirmed that these strains of V. cholerae did not utilize malonate as a primary carbon source. However, we found that the addition of malonate to the growth medium stimulated V. cholerae growth. All together, these results suggest that metabolizing malonate as a nutrient source negatively affects virulence gene expression in V. cholerae.

  9. 产色素O139群霍乱弧菌环境分离株的亲缘关系分析%Clonality of pigment producing O139 Vibrio cholerae strains isolated in outer environment

    Institute of Scientific and Technical Information of China (English)

    王瑞白; 阚飙

    2012-01-01

    Objective To screen the O139 serogroup Vibrio cholerae strains that can produce brown pigment and analyze the clonality of these pigment producing strains. Methods A total of 1530 strains of O139 V. cholerae were detected to understand their ability to produce soluble brown pigment. Their homogentisate oxygenase genes ( VC1345) were sequenced and their clonality relation was analyzed by PFGE typing and MLST. Results Sixteen strains of pigment producing but non-toxin producing 0139 V. cholerae strains isolated in 3 provinces in 13 years were selected. They had completely same VC1345 gene, highly similar PFGE type, and two kinds of MLST types. Conclusion All of the pigment producing O139 V. cholera strains isolated in outer environment came from one phylogenetic clone group with low degree diversity. The sustained existence of such strains in outer environment indicates that they may have strong survival ability, suggesting that close attention should be paid to such strains in the surveillance of V. cholerae.%目的 筛检霍乱弧菌O139血清群产褐色色素的环境分离菌株并分析其遗传近缘关系.方法 在LBA平板上检测了1992 -2010年分离保存的全国1530株O139群霍乱弧菌的色素产生能力,对其中能够产生水溶性褐色色素的菌株的马尿酸氧化酶基因VC1345进行测序,并用脉冲场凝胶电泳(PFGE)、多基因序列分型分析色素产生菌株之间的亲缘关系.结果 筛选到不同年份不同地区的16株产色素的非产毒O139群菌株.所有的色素产生菌株具有完全一致的VC 1345基因、相似的PFGE带型,多基因序列分型显示仅有2种型别.结论 分离到的O139群产色素菌株来源于近缘的克隆群,而这些菌株之间已存在一定程度的变异.在水体中的持续存在显示其较强的环境生存能力,这些特征提示在霍乱弧菌的环境监测中需提高对这类菌株的关注.

  10. Proteolysis of virulence regulator ToxR is associated with entry of Vibrio cholerae into a dormant state.

    Directory of Open Access Journals (Sweden)

    Salvador Almagro-Moreno

    2015-04-01

    Full Text Available Vibrio cholerae O1 is a natural inhabitant of aquatic environments and causes the diarrheal disease, cholera. Two of its primary virulence regulators, TcpP and ToxR, are localized in the inner membrane. TcpP is encoded on the Vibrio Pathogenicity Island (VPI, a horizontally acquired mobile genetic element, and functions primarily in virulence gene regulation. TcpP has been shown to undergo regulated intramembrane proteolysis (RIP in response to environmental conditions that are unfavorable for virulence gene expression. ToxR is encoded in the ancestral genome and is present in non-pathogenic strains of V. cholerae, indicating it has roles outside of the human host. In this study, we show that ToxR undergoes RIP in V. cholerae in response to nutrient limitation at alkaline pH, a condition that occurs during the stationary phase of growth. This process involves the site-2 protease RseP (YaeL, and is dependent upon the RpoE-mediated periplasmic stress response, as deletion mutants for the genes encoding these two proteins cannot proteolyze ToxR under nutrient limitation at alkaline pH. We determined that the loss of ToxR, genetically or by proteolysis, is associated with entry of V. cholerae into a dormant state in which the bacterium is normally found in the aquatic environment called viable but nonculturable (VBNC. Strains that can proteolyze ToxR, or do not encode it, lose culturability, experience a change in morphology associated with cells in VBNC, yet remain viable under nutrient limitation at alkaline pH. On the other hand, mutant strains that cannot proteolyze ToxR remain culturable and maintain the morphology of cells in an active state of growth. Overall, our findings provide a link between the proteolysis of a virulence regulator and the entry of a pathogen into an environmentally persistent state.

  11. Optimization and head-to-head comparison of MISSR-PCR, ERIC-PCR, RAPD and 16S rRNA evolutionary clock for the genotyping of Vibrio cholerae isolated in China

    Directory of Open Access Journals (Sweden)

    Q H Mo

    2015-01-01

    Full Text Available Purpose: To establish a new genotyping method for Vibrio cholerae and compare it with other methods. Materials and Methods: In the current study, a modified inter simple sequence repeat-polymerase chain reaction (MISSR-PCR system was developed via several rounds of optimisation. Comparison study was then conducted between MISSR-PCR and three other methods, including enterobacterial repetitive intergenic consensus sequences-based PCR (ERIC-PCR, randomly amplified polymorphic DNA (RAPD and 16S rRNA evolutionary clock, for the detection and genetic tracing of Vibrio cholerae isolated from seafood in China. Result: The results indicated that the MISSR-PCR system could generate the highest polymorphic fingerprinting map in a single round PCR and showed the best discriminatory ability for Vibrio cholerae genotyping by clearly separating toxigenic/nontoxigenic strains, local/foreign strains, and O1/O139/non-O1/non-O139 serogroup strains, comparing to ERIC-PCR, RAPD and 16S rRNA evolutionary clock. Moreover, the MISSR-PCR is superior to previously described traditional simple sequence repeat based PCR method on genotyping by more clearly separating different clusters. Conclusion: To the best of our knowledge, this is the first head-to-head comparison of four detection and genotyping methods for Vibrio cholerae The MISSR-PCR system established here could serve as a simple, quick, reliable and cost-effective tool for the genotyping and epidemiological study.

  12. Molecular characteristics and antibiotic resistance of Vibrio cholerae O139 in Shandong province

    Institute of Scientific and Technical Information of China (English)

    袁玉起

    2014-01-01

    Objective To investigate the molecular epidemiological characteristics and antibiotic resistance profiles of Vibrio cholerae O139 in Shandong province.Methods A total of 13 strains of V.cholerae O139(9 clinical strains and 4 environmental strains)isolated from cholera epidemics in Shandong province since 1997 were recovered and confirmed with serum agglutination and biochemical reaction.Pulsed-field gel electrophoresis(PFGE)was carried out for molecular subtyping.Virulence genes and

  13. Analysis of Vibrio cholerae toxR function by construction of toxR gene deficient strains of Vibrio cholerae%霍乱弧菌毒力表达调控基因toxR缺失株的构建及其功能的研究

    Institute of Scientific and Technical Information of China (English)

    陈建平; 高守一; 刘延清; 祁国明; 何九芽

    2001-01-01

    Objective To analyze toxR gene function of Vibrio cholerae byconstruction of toxR gene deficient strains of Vibrio cholerae IEM101-4 and 569B-43. Methods A 2.5kb tetracycline gene fragment was homo-recombined into a toxR gene site in genome of Vibrio cholerae IEM101 and 569B using suicide plasmid pGp704 and integrative recombination method, and constructed 2 toxR gene deficient strains of Vibrio cholerae IEM101-4 and 569B-43. We analyzed the CT production and outer membrane protein of toxR gene deficient strains of Vibrio cholerae and donor strains. Results GM1-ELISA assay of CT toxin of Vibrio cholerae showed that the level of CT production of the toxR gene deficient strain, 569B-43 was much lower than that of donor strain 569B, the P/N value of toxR gene deficient strain 569B-43 was 1.82, the P/N value of 569B was 4.52. No difference was found between IEM101 and the toxR gene deficient strains IEM101-4 in CT production. SDS-PAGE analysis of outer membrane protein of Vibrio cholerae showed that the toxR gene deficient strains, IEM101-4 and 569B-43 all showed 40kD and 43kD outer membrane protein bands, while which were not present in the donor strains IEM101 and 569B. Conclusion ToxR gene is an up-regulator for the expression of ctx gene, and a down-regulator for the expression of 40kD and 43kD outer membrane protein coding genes.%目的 通过构建霍乱弧菌toxR基因缺失株来研究toxR基因对霍乱弧菌减毒菌株IEM101和高产毒株569B毒力表达的调控作用。方法 采用自杀性质粒和接合转移技术,将2个中间含有四环素基因的toxR基因分别与霍乱弧菌减毒株IEM101和高产毒株569B染色体toxR基因重组,从而获得toxR基因缺失株IEM101-4和569B-43,并对2个toxR基因缺失株和其原出发菌株的霍乱肠毒素的产率和主要外膜蛋白图谱进行比较。结果 采用GM1-ELISA检测受测菌CT基因表达,toxR基因缺失株569B-43的P/N值为1.82,而其原出发菌株569B的P/N为4.52

  14. Cholera epidemic threatens Sierra Leone.

    Science.gov (United States)

    Dyer, O

    1995-07-08

    Sierra Leone faces the threat of a major epidemic of cholera with the onset of the rainy season, according to the World Health Organization (WHO). The situation is particularly grave for the two million people displaced by the country's civil war. Already 1709 cases of cholera have been registered in Freetown, with 57 deaths. Freetown's population has doubled since the start of the war in 1991 with 750,000 refugees camping out in the town. The insurgent Revolutionary United Front is now within 32 km of the capital. Provinces are cut off from the capital, medical supplies are scarce. Doctors and aid workers are forced to rely on a private helicopter service for personal transport. As many as 10,000 people were affected by the disease last year. WHO experts predict that pneumonia is likely to claim the lives of many children, and a highly drug resistant strain of Plasmodium falciparum malaria is also looming. The greatest problems are the lack of safe drinking water and the attendant risks of cholera and dysentery. At one site in Freetown the 6000 refugees have to fetch water from a well and have no latrines. As a result there have been 277 cases of cholera and 2 deaths already among that group. The health department has set up five centers to treat cholera in Freetown and is organizing mobile clinics. WHO's Sierra Leone office is assisting the government mobile health teams, which provide free primary care to displaced people. Medicines and vaccines, however, are lacking. Many of the staff of the 13 district health authorities have been displaced to Freetown. Aid agencies such as Medecins Sans Frontieres and Oxfam have stepped into the role in many districts. Ironically, one of the Revolutionary United Front's main demands is for a free national health service.

  15. The Vibrio cholerae type VI secretion system employs diverse effector modules for intraspecific competition.

    Science.gov (United States)

    Unterweger, Daniel; Miyata, Sarah T; Bachmann, Verena; Brooks, Teresa M; Mullins, Travis; Kostiuk, Benjamin; Provenzano, Daniele; Pukatzki, Stefan

    2014-04-01

    Vibrio cholerae is a Gram-negative bacterial pathogen that consists of over 200 serogroups with differing pathogenic potential. Only strains that express the virulence factors cholera toxin (CT) and toxin-coregulated pilus (TCP) are capable of pandemic spread of cholera diarrhoea. Regardless, all V. cholerae strains sequenced to date harbour genes for the type VI secretion system (T6SS) that translocates effectors into neighbouring eukaryotic and prokaryotic cells. Here we report that the effectors encoded within these conserved gene clusters differ widely among V. cholerae strains, and that immunity proteins encoded immediately downstream from the effector genes protect their host from neighbouring bacteria producing corresponding effectors. As a consequence, strains with matching effector-immunity gene sets can coexist, while strains with different sets compete against each other. Thus, the V. cholerae T6SS contributes to the competitive behaviour of this species.

  16. Time series analysis of cholera in Matlab, Bangladesh, during 1988-2001.

    Science.gov (United States)

    Ali, Mohammad; Kim, Deok Ryun; Yunus, Mohammad; Emch, Michael

    2013-03-01

    The study examined the impact of in-situ climatic and marine environmental variability on cholera incidence in an endemic area of Bangladesh and developed a forecasting model for understanding the magnitude of incidence. Diarrhoea surveillance data collected between 1988 and 2001 were obtained from a field research site in Matlab, Bangladesh. Cholera cases were defined as Vibrio cholerae O1 isolated from faecal specimens of patients who sought care at treatment centres serving the Matlab population. Cholera incidence for 168 months was correlated with remotely-sensed sea-surface temperature (SST) and in-situ environmental data, including rainfall and ambient temperature. A seasonal autoregressive integrated moving average (SARIMA) model was used for determining the impact of climatic and environmental variability on cholera incidence and evaluating the ability of the model to forecast the magnitude of cholera. There were 4,157 cholera cases during the study period, with an average of 1.4 cases per 1,000 people. Since monthly cholera cases varied significantly by month, it was necessary to stabilize the variance of cholera incidence by computing the natural logarithm to conduct the analysis. The SARIMA model shows temporal clustering of cholera at one- and 12-month lags. There was a 6% increase in cholera incidence with a minimum temperature increase of one degree celsius in the current month. For increase of SST by one degree celsius, there was a 25% increase in the cholera incidence at currrent month and 18% increase in the cholera incidence at two months. Rainfall did not influenc to cause variation in cholera incidence during the study period. The model forecast the fluctuation of cholera incidence in Matlab reasonably well (Root mean square error, RMSE: 0.108). Thus, the ambient and sea-surface temperature-based model could be used in forecasting cholera outbreaks in Matlab.

  17. 利用脉冲场凝胶电泳及核糖体分型技术对14株上海崇明地区O139群霍乱弧菌分型分析%Study on 14 Vibrio cholerae O139 strains by PFGE and ribotyping in Chongming, Shanghai

    Institute of Scientific and Technical Information of China (English)

    屠丽红; 陈洪友; 陈敏

    2011-01-01

    目的 对14株上海市崇明地区2005年O139群霍乱弧菌菌株进行脉冲场凝胶电泳(PFCE)及核糖体分型分析.方法 用PFGE、核糖体分型对霍乱弧菌进行分析.结果 14株O139群霍乱弧菌产毒株经核糖体分型分析为同一型;而PFGE双酶切分型结果为其中10株为同一型,其余4株为高度同源性.结论 上海市崇明地区2005年流行的O139群霍乱弧菌核酸指纹分型基本一致.%Objective To understand the genotypes of 14 strains of Vibrio cholerae O139 isolated in Chongming in 2005. Methods Pulsed-field gel electrophoresis (PFGE) and ribotyping were conducted to analyze the Vibrio cholerae strains. Results Ribotyping indicated that the genotypes of all the O139 strains were same; but PFGE indicated that 10 of 14 strains of Vibrio cholerae O139 belonged to same type, the other 4 strains were highly homologous. Conclusion The genotypes of Vibrro cholerae O139 circulating in Chongming in Shanghai in 2005 were similarly.

  18. Cholera risk factors, Papua New Guinea, 2010

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    Rosewell Alexander

    2012-11-01

    Full Text Available Abstract Background Cholera is newly emergent in Papua New Guinea but may soon become endemic. Identifying the risk factors for cholera provides evidence for targeted prevention and control measures. Methods We conducted a hospital-based case–control study to identify cholera risk factors. Using stool culture as the standard, we evaluated a cholera point of care test in the field. Results 176 participants were recruited: 54 cases and 122 controls. Independent risk factors for cholera were: being over 20 years of age (aOR 2.5; 95%CI 1.1, 5.4, defecating in the open air (or river (aOR 4.5; 95% CI 1.4, 14.4 and knowing someone who travelled to a cholera affected area (aOR 4.1; 95%CI 1.6, 10.7; while the availability of soap for handwashing at home was protective (aOR 0.41; 95%CI 0.19, 0.87. Those reporting access to a piped water distribution system in the home were twice as likely to report the availability of soap for handwashing. The sensitivity and specificity of the rapid test were 72% (95% CI 47–90 and 71% (95%CI 44–90%. Conclusions Improving population access to the piped water distribution system and sanitation will likely reduce transmission by enabling enhanced hygiene and limiting the contamination of water sources. The One step V. cholerae O1/O139 Antigen Test is of limited utility for clinical decision making in a hospital setting with access to traditional laboratory methods. Settlement dwellers and mobile populations of all age groups should be targeted for interventions in Papua New Guinea.

  19. [DETERMINATION OF TYPES OF EPIDEMIC MANIFESTATIONS OF CHOLERA IN REGIONS OF THE CRIMEA FEDERAL DISTRICT (REPUBLIC OF CRIMEA)].

    Science.gov (United States)

    Onischenko, G G; Popova, A Yu; Moskvitina, E A; Penkovskaya, N A; Listopad, S A; Titova, S V; Kruglikov, V D

    2015-01-01

    The aim of the study was determination of the type of epidemic manifestations of cholera in the Republic of Crimea based on evaluation of epidemic manifestations of cholera risk of introduction and spread of the infection. It was concluded, that, based on the cholera outbreaks, that had taken place, contamination of surface water bodies (fresh and sea) and sewage by Vibrio cholerae O1 ctxA+ and Vibrio cholerae O1 ctXA- potential epidemic danger of introduction of the infection by various types of international transport, population migration, the presence of epidemiologic risk in realization of water pathway of transmission of cholera causative agent and several other social conditions, the Republic of Crimea remains in the group of territories of type I by epidemic manifestations of cholera.

  20. [Isolation of the R'his plasmids of Vibrio cholerae].

    Science.gov (United States)

    Rusina, O Iu; Tiganova, I G; Aleshkin, G I; Andreeva, I V; Skavronskaia, A G

    1987-06-01

    V. cholerae strain VT5104 capable of donor activity in conjugation has been constructed by the genetic technique based on plasmid RP4::Mucts62 integration into V. cholerae chromosome due to plasmid homology with Mucts62 inserted into the chromosome. The gene for histidine synthesis has been mobilized and transferred into the recipient cells from VT5104 donor. The conjugants obtained are able to efficiently transfer his+ gene included into the plasmid structure in conjugation with eltor recipient. Thus, the constructed strain VT5104 generates R' plasmids carrying V. cholerae chromosomal genes.

  1. Vibrio cholerae Biofilms and Cholera Pathogenesis.

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    Anisia J Silva

    2016-02-01

    Full Text Available Vibrio cholerae can switch between motile and biofilm lifestyles. The last decades have been marked by a remarkable increase in our knowledge of the structure, regulation, and function of biofilms formed under laboratory conditions. Evidence has grown suggesting that V. cholerae can form biofilm-like aggregates during infection that could play a critical role in pathogenesis and disease transmission. However, the structure and regulation of biofilms formed during infection, as well as their role in intestinal colonization and virulence, remains poorly understood. Here, we review (i the evidence for biofilm formation during infection, (ii the coordinate regulation of biofilm and virulence gene expression, and (iii the host signals that favor V. cholerae transitions between alternative lifestyles during intestinal colonization, and (iv we discuss a model for the role of V. cholerae biofilms in pathogenicity.

  2. Modeling cholera outbreaks

    OpenAIRE

    Dennis L Chao; Longini, Ira M.; Morris, J. Glenn

    2014-01-01

    Mathematical modeling can be a valuable tool for studying infectious disease outbreak dynamics and simulating the effects of possible interventions. Here, we describe approaches to modeling cholera outbreaks and how models have been applied to explore intervention strategies, particularly in Haiti. Mathematical models can play an important role in formulating and evaluating complex cholera outbreak response options. Major challenges to cholera modeling are insufficient data for calibrating mo...

  3. Regulation by ToxR-Like Proteins Converges on vttRB Expression To Control Type 3 Secretion System-Dependent Caco2-BBE Cytotoxicity in Vibrio cholerae

    OpenAIRE

    Miller, Kelly A.; Sofia, Madeline K.; Weaver, Jacob W. A.; Christopher H. Seward; Dziejman, Michelle

    2016-01-01

    Genes carried on the type 3 secretion system (T3SS) pathogenicity island of Vibrio cholerae non-O1/non-O139 serogroup strain AM-19226 must be precisely regulated in order for bacteria to cause disease. Previously reported results showed that both T3SS function and the presence of bile are required to cause Caco2-BBE cell cytotoxicity during coculture with strain AM-19226. We therefore investigated additional parameters affecting in vitro cell death, including bacterial load and the role of th...

  4. Vibrio cholerae VttRA and VttRB Regulatory Influences Extend beyond the Type 3 Secretion System Genomic Island

    OpenAIRE

    Chaand, Mudit; Dziejman, Michelle

    2013-01-01

    A subset of non-O1/non-O139 serogroup strains of Vibrio cholerae cause disease using type 3 secretion system (T3SS)-mediated mechanisms. An ∼50-kb genomic island carries genes encoding the T3SS structural apparatus, effector proteins, and two transmembrane transcriptional regulators, VttRA and VttRB, which are ToxR homologues. Previous experiments demonstrated that VttRA and VttRB are necessary for colonization in vivo and promote bile-dependent T3SS gene expression in vitro. To better unders...

  5. Dual expression profile of type VI secretion system immunity genes protects pandemic Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    Sarah T Miyata

    Full Text Available The Vibrio cholerae type VI secretion system (T6SS assembles as a molecular syringe that injects toxic protein effectors into both eukaryotic and prokaryotic cells. We previously reported that the V. cholerae O37 serogroup strain V52 maintains a constitutively active T6SS to kill other Gram-negative bacteria while being immune to attack by kin bacteria. The pandemic O1 El Tor V. cholerae strain C6706 is T6SS-silent under laboratory conditions as it does not produce T6SS structural components and effectors, and fails to kill Escherichia coli prey. Yet, C6706 exhibits full resistance when approached by T6SS-active V52. These findings suggested that an active T6SS is not required for immunity against T6SS-mediated virulence. Here, we describe a dual expression profile of the T6SS immunity protein-encoding genes tsiV1, tsiV2, and tsiV3 that provides pandemic V. cholerae strains with T6SS immunity and allows T6SS-silent strains to maintain immunity against attacks by T6SS-active bacterial neighbors. The dual expression profile allows transcription of the three genes encoding immunity proteins independently of other T6SS proteins encoded within the same operon. One of these immunity proteins, TsiV2, protects against the T6SS effector VasX which is encoded immediately upstream of tsiV2. VasX is a secreted, lipid-binding protein that we previously characterized with respect to T6SS-mediated virulence towards the social amoeba Dictyostelium discoideum. Our data suggest the presence of an internal promoter in the open reading frame of vasX that drives expression of the downstream gene tsiV2. Furthermore, VasX is shown to act in conjunction with VasW, an accessory protein to VasX, to compromise the inner membrane of prokaryotic target cells. The dual regulatory profile of the T6SS immunity protein-encoding genes tsiV1, tsiV2, and tsiV3 permits V. cholerae to tightly control T6SS gene expression while maintaining immunity to T6SS activity.

  6. Cholera in coastal Africa: a systematic review of its heterogeneous environmental determinants.

    Science.gov (United States)

    Rebaudet, Stanislas; Sudre, Bertrand; Faucher, Benoît; Piarroux, Renaud

    2013-11-01

    According to the "cholera paradigm," epidemiology of this prototypical waterborne disease is considered to be driven directly by climate-induced variations in coastal aquatic reservoirs of Vibrio cholerae. This systematic review on environmental determinants of cholera in coastal Africa shows that instead coastal epidemics constitute a minor part of the continental cholera burden. Most of coastal cholera foci are located near estuaries, lagoons, mangrove forests, and on islands. Yet outbreaks often originate in coastal cities, where cholera is more likely to be imported from distant areas. Cholera outbreaks also may intensify in densely populated slum quarters before spreading to adjacent regions. Frequent seasonality of cholera incidence appears driven by the rainfall-induced contamination of unprotected water sources through latrine overflow and sewage, as well as by the periodicity of human activities like fishing or traveling. Lulls in transmission periods of several years are repeatedly recorded even in high-risk coastal areas. To date, environmental studies have failed to demonstrate a perennial aquatic reservoir of toxigenic V. cholerae around the continent. Finally, applicability of the cholera paradigm therefore appears questionable in Africa, although available data remain limited. Thorough surveys with microbiological analyses of water samples and prospective genotyping of environmental and clinical strains of V. cholerae are needed to understand determinants of cholera in coastal Africa and better target prevention and control measures.

  7. Cholera outbreaks in India.

    Science.gov (United States)

    Ramamurthy, Thandavarayan; Sharma, Naresh C

    2014-01-01

    Cholera is a global health problem as several thousands of cases and deaths occur each year. The unique epidemiologic attribute of the disease is its propensity to occur as outbreaks that may flare-up into epidemics, if not controlled. The causative bacterial pathogen Vibrio cholerae prevails in the environment and infects humans whenever there is a breakdown in the public health component. The Indian subcontinent is vulnerable to this disease due its vast coastlines with areas of poor sanitation, unsafe drinking water, and overcrowding. Recently, it was shown that climatic conditions also play a major role in the persistence and spread of cholera. Constant change in the biotypes and serotypes of V. cholerae are also important aspects that changes virulence and survival of the pathogen. Such continuous changes increase the infection ability of the pathogen affecting the susceptible population including the children. The short-term carrier status of V. cholerae has been studied well at community level and this facet significantly contributes to the recurrence of cholera. Several molecular tools recognized altering clonality of V. cholerae in relation with the advent of a serogroup or serotype. Rapid identification systems were formulated for the timely detection of the pathogen so as to identify and control the outbreak and institute proper treatment of the patients. The antimicrobials used in the past are no longer useful in the treatment of cholera as V. cholerae has acquired several mechanisms for multiple antimicrobial resistance. This upsurge in antimicrobial resistance directly influences the management of the disease. This chapter provides an overview of cholera prevalence in India, possible sources of infection, and molecular epidemiology along with antimicrobial resistance of V. cholerae.

  8. Origins of the current seventh cholera pandemic.

    Science.gov (United States)

    Hu, Dalong; Liu, Bin; Feng, Lu; Ding, Peng; Guo, Xi; Wang, Min; Cao, Boyang; Reeves, Peter R; Wang, Lei

    2016-11-29

    Vibrio cholerae has caused seven cholera pandemics since 1817, imposing terror on much of the world, but bacterial strains are currently only available for the sixth and seventh pandemics. The El Tor biotype seventh pandemic began in 1961 in Indonesia, but did not originate directly from the classical biotype sixth-pandemic strain. Previous studies focused mainly on the spread of the seventh pandemic after 1970. Here, we analyze in unprecedented detail the origin, evolution, and transition to pandemicity of the seventh-pandemic strain. We used high-resolution comparative genomic analysis of strains collected from 1930 to 1964, covering the evolution from the first available El Tor biotype strain to the start of the seventh pandemic. We define six stages leading to the pandemic strain and reveal all key events. The seventh pandemic originated from a nonpathogenic strain in the Middle East, first observed in 1897. It subsequently underwent explosive diversification, including the spawning of the pandemic lineage. This rapid diversification suggests that, when first observed, the strain had only recently arrived in the Middle East, possibly from the Asian homeland of cholera. The lineage migrated to Makassar, Indonesia, where it gained the important virulence-associated elements Vibrio seventh pandemic island I (VSP-I), VSP-II, and El Tor type cholera toxin prophage by 1954, and it then became pandemic in 1961 after only 12 additional mutations. Our data indicate that specific niches in the Middle East and Makassar were important in generating the pandemic strain by providing gene sources and the driving forces for genetic events.

  9. Phenotypic and Genetic Heterogeneity in Vibrio cholerae O139 Isolated from Cholera Cases in Delhi, India during 2001-2006.

    Science.gov (United States)

    Ghosh, Raikamal; Sharma, Naresh C; Halder, Kalpataru; Bhadra, Rupak K; Chowdhury, Goutam; Pazhani, Gururaja P; Shinoda, Sumio; Mukhopadhyay, Asish K; Nair, G Balakrish; Ramamurthy, Thadavarayan

    2016-01-01

    Incidence of epidemic Vibrio cholerae serogroup O139 has declined in cholera endemic countries. However, sporadic cholera caused by V. cholerae O139 with notable genetic changes is still reported from many regions. In the present study, 42 V. cholerae O139 strains isolated from 2001 to 2006 in Delhi, India, were retrospectively analyzed to understand their phenotype and molecular characteristics. The majority of isolates were resistant to ampicillin, furazolidone and nalidixic acid. Though the integrative conjugative element was detected in all the O139 isolates, the 2004-2006 isolates remained susceptible to co-trimoxazole, chloramphenicol, and streptomycin. Cholera toxin genotype 1 was present in the majority of the O139 isolates while few had type 3 or a novel type 4. In the cholera toxin encoding gene (ctx) restriction fragment length polymorphism, the majority of the isolates harbored three copies of CTX element, of which one was truncated. In this study, the ctx was detected for the first time in the small chromosome of V. cholerae O139 and one isolate harbored 5 copies of CTX element, of which 3 were truncated. The ribotype BII pattern was found in most of the O139 isolates. Three V. cholerae O139 isolated in 2001 had a new ribotype BVIII. Pulsed-field gel electrophoresis analysis revealed clonal variation in 2001 isolates compared to the 2004-2006 isolates. Molecular changes in V. cholerae O139 have to be closely monitored as this information may help in understanding the changing genetic features of this pathogen in relation to the epidemiology of cholera.

  10. Phenotypic and Genetic Heterogeneity in Vibrio cholerae O139 Isolated from Cholera Cases in Delhi, India during 2001-2006

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    Raikamal Ghosh

    2016-08-01

    Full Text Available Incidence of epidemic Vibrio cholerae serogroup O139 has declined in cholera endemic countries. However, sporadic cholera caused by V. cholerae O139 with notable genetic changes is still reported from many regions. In the present study, 42 V. cholerae O139 strains isolated from 2001 to 2006 in Delhi, India, were retrospectively analyzed to understand their phenotype and molecular characteristics. The majority of isolates were resistant to ampicillin, furazolidone and nalidixic acid. Though the integrative conjugative element was detected in all the O139 isolates, the 2004-2006 isolates remained susceptible to co-trimoxazole, chloramphenicol, and streptomycin. Cholera toxin genotype 1 was present in the majority of the O139 isolates while few had type 3 or a novel type 4. In the cholera toxin encoding gene (ctx restriction fragment length polymorphism, the majority of the isolates harbored three copies of CTX element, of which one was truncated. In this study, the ctx was detected for the first time in the small chromosome of V. cholerae O139 and one isolate harbored 5 copies of CTX element, of which 3 were truncated. The ribotype BII pattern was found in most of the O139 isolates. Three V. cholerae O139 isolated in 2001 had a new ribotype BVIII. Pulsed-field gel electrophoresis analysis revealed clonal variation in 2001 isolates compared to the 2004-2006 isolates. Molecular changes in V. cholerae O139 have to be closely monitored as this information may help in understanding the changing genetic features of this pathogen in relation to the epidemiology of cholera.

  11. Synthesis of the methyl alpha-glycosides of a di-, tri-, and a tetra-saccharide fragment mimicking the terminus of the O-polysaccharide of Vibrio cholerae O:1, serotype Ogawa.

    Science.gov (United States)

    Lei, P; Ogawa, Y; Kovác, P

    1996-02-07

    Methyl 4-(3-deoxy-L-glycero-tetronamido)-4,6-dideoxy-2-O-methyl-alpha-D- mannopyranoside was acetylated, and the fully protected methyl glycoside was treated with dichloromethyl methyl ether-ZnCl2 (DCMME-ZnCl2) reagent to give 3-O-acetyl-4-(2,4-di-O-acetyl-3- deoxy-L-glycero-tetronamido)-4,6-dideoxy-2-O-methyl-alpha-D-mannop yranosyl chloride (3). Condensation of 3 with methyl 3-O-acetyl-4-(2,4-di-O-acetyl-3-deoxy-L-glycero-tetronamido)-4,6- dideoxy-alpha-D-mannopyranoside (4) gave the fully acetylated disaccharide 5, which was deacetylated yielding the methyl alpha-glycoside of title disaccharide. The disaccharide glycosyl donor required for the blockwise synthesis of the title tri- and the tetra-saccharide, 3-O-acetyl-4-(2,4-di-O-acetyl-3-deoxy-L-glycero-tetronamido)-4,6-d ideoxy-2-O- methyl-alpha-D-mannopyranosyl-(1-->2)-3-O-acetyl-4- (2,4-di-O-acetyl-3-deoxy-L-glycero-tetronamido)-4,6-dideoxy-alpha- D- mannopyranosyl chloride (12), was obtained by condensation of 3 with the 1-O-acetyl analog of 4, followed by treatment of the disaccharide formed with DCMME-ZnCl2. The synthesis of the methyl alpha-glycoside of the title trisaccharide involved a condensation of 12 with 4, followed by deacetylation. Similarly, the condensation of 12 with 15, the latter being the analog of 5 having a free HO-2, followed by deacetylation, gave the methyl alpha-glycoside of the title tetrasaccharide. All glycosylation reactions were mediated by silver trifluoromethanesulfonate in the presence of 2,4,6-trimethylpyridine. 4-(3-Deoxy-L-glycero-tetronamido)-4,6-dideoxy-2-O-methyl-alpha,bet a-D- mannopyranose was prepared for the first time. It was characterized by NMR spectroscopy, and via its crystalline per-O-acetyl derivative. It is the saccharide whose alpha-form constitutes the terminal, non-reducing end-group of the O-PS of V. cholerea O:1, serotype Ogawa.

  12. Survival and proliferation of the lysogenic bacteriophage CTXΦ in Vibrio cholerae

    Institute of Scientific and Technical Information of China (English)

    Fenxia; Fan; Biao; Kan

    2015-01-01

    The lysogenic phage CTXΦ of Vibrio cholerae can transfer the cholera toxin gene both horizontally(inter-strain) and vertically(cell proliferation). Due to its diversity in form and species, the complexity of regulatory mechanisms, and the important role of the infection mechanism in the production of new virulent strains of V.cholerae, the study of the lysogenic phage CTXΦ has attracted much attention. Based on the progress of current research, the genomic features and their arrangement, the host-dependent regulatory mechanisms of CTXΦ phage survival, proliferation and propagation were reviewed to further understand the phage’s role in the evolutionary and epidemiological mechanisms of V. cholerae.

  13. Removal of Cholera Toxin from Aqueous Solution by Probiotic Bacteria

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    Jussi A. O. Meriluoto

    2012-06-01

    Full Text Available Cholera remains a serious health problem, especially in developing countries where basic hygiene standards are not met. The symptoms of cholera are caused by cholera toxin, an enterotoxin, which is produced by the bacterium Vibrio cholerae. We have recently shown that human probiotic bacteria are capable of removing cyanobacterial toxins from aqueous solutions. In the present study we investigate the ability of the human probiotic bacteria, Lactobacillus rhamnosus strain GG (ATCC 53103 and Bifidobacterium longum 46 (DSM 14583, to remove cholera toxin from solution in vitro. Lactobacillus rhamnosus strain GG and Bifidobacterium longum 46 were able to remove 68% and 59% of cholera toxin from aqueous solutions during 18 h of incubation at 37 °C, respectively. The effect was dependent on bacterial concentration and L. rhamnosus GG was more effective at lower bacterial concentrations. No significant effect on cholera toxin concentration was observed when nonviable bacteria or bacterial supernatant was used.

  14. A study on the geophylogeny of clinical and environmental Vibrio cholerae in Kenya.

    Directory of Open Access Journals (Sweden)

    John Kiiru

    Full Text Available Cholera remains a significant public health challenge in many sub-Saharan countries including Kenya. We have performed a combination of phylogenetic and phenotypic analysis based on whole genome DNA sequences derived from 40 environmental and 57 clinical V. cholerae from different regions of Kenya isolated between 2005 and 2010. Some environmental and all clinical isolates mapped back onto wave three of the monophyletic seventh pandemic V. cholerae El Tor phylogeny but other environmental isolates were phylogenetically very distinct. Thus, the genomes of the Kenyan V. cholerae O1 El Tor isolates are clonally related to other El Tor V. cholerae isolated elsewhere in the world and similarly harbour antibiotic resistance-associated STX elements. Further, the Kenyan O1 El Tor isolates fall into two distinct clades that may have entered Kenya independently.

  15. Accessory cholera enterotoxin, Ace, from Vibrio cholerae: structure, unfolding, and virstatin binding.

    Science.gov (United States)

    Chatterjee, Tanaya; Mukherjee, Debadrita; Dey, Sucharita; Pal, Aritrika; Hoque, Kazi Mirajul; Chakrabarti, Pinak

    2011-04-12

    Vibrio cholerae accessory cholera enterotoxin (Ace) is the third toxin, along with cholera toxin (CT) and zonula occludens toxin (Zot), that causes the endemic disease cholera. Structural characterization of Ace has been restricted because of the limited production of this toxic protein by V. cholerae. We have cloned, overexpressed, and purified Ace from V. cholerae strain O395 in Escherichia coli to homogeneity and determined its biological activity. The unfolding of the purified protein was investigated using circular dichroism and intrinsic tryptophan fluorescence. Because Ace is predominantly a hydrophobic protein, the degree of exposure of hydrophobic regions was identified from the spectral changes of the environment-sensitive fluorescent probe 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (bis-ANS) that quenches the fluorescence of tryptophan residues of Ace in a concentration-dependent manner. Results showed that bis-ANS binds one monomeric unit of Ace with a 1:1 stoichiometry and a K' of 0.72 μM. Ace exists as a dimer, with higher oligomeric forms appearing upon glutaraldehyde cross-linking. This study also reports the binding of virstatin, a small molecule that inhibits virulence regulation in V. cholerae, to Ace. The binding constant (K=9×10(4) M(-1)) and the standard free energy change (ΔG°=-12 kcal mol(-1)) of Ace-virstatin interaction have been evaluated by the fluorescence quenching method. The binding does not affect the oligomeric status of Ace. A cell viability assay of the antibacterial activity of Ace has been performed using various microbial strains. A homology model of Ace, consistent with the experimental results, has been constructed.

  16. Radiolabelling of cholera toxin

    Energy Technology Data Exchange (ETDEWEB)

    Santos, R.G.; Neves, Nicoli M.J. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN), Belo Horizonte, MG (Brazil); Abdalla, L.F.; Brandao, R.L.; Etchehebehere, L. [Ouro Preto Univ., MG (Brazil). Escola de Farmacia. Lab. de Fisiologia e Bioquimica de Microorganismos; Lima, M.E. de [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Bioquimica e Imunologia; Nicoli, J.R. [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Microbiologia

    1999-11-01

    Binding of cholera toxin to ganglioside receptors of enterocyte microvilli catalyzes the activation of adenylate cyclase causing a rise in cAMP which final result is a copious diarrhea. Saccharomyces boulardii, a nonpathogenic yeast has been used to prevent diarrhea. Although the antidiarrheic properties of S. boulardii are widely recognized, this yeast has been used on empirical basis, and the mechanism of this protective effect is unknown. The addition of cholera toxin to S. boulardii induces the raising of cAMP that triggers the activation of neutral trehalase. This suggests that toxin specifically binding to cells, is internalized and active the protein phosphorylation cascade. Our objective is labeling the cholera toxin to verify the presence of binding sites on yeast cell surfaces for the cholera toxin. Cholera toxin was radiolabelled with Na {sup 125} I by a chloramine-T method modified from Cuatrecasas and Griffiths et alii. The {sup 125} I-Cholera toxin showed a specific radioactivity at about 1000 cpm/fmol toxin. Biological activity of labeled cholera toxin measured by trehalase activation was similar to the native toxin. (author) 5 refs., 3 figs.; e-mail: nevesmj at urano.cdtn.br

  17. Biological characterization of v. Cholerae-specific bacteriophages isolated from water sources in Georgia.

    Science.gov (United States)

    Elbakidze, T; Kokashvili, T; Janelidze, N; Porchkhidze, K; Koberidze, T; Tediashvili, M

    2015-03-01

    Vibrio cholerae, a widely spread bacterium in various marine, fresh, and brackish water environments, can cause a devastating diarrheal disease - cholera and also mild forms of gastroenteritis. Bacterial viruses are natural controllers of bacterial population density in water systems. The goal of this study was to isolate and characterize V. cholerae-specific bacteriophages occurring in the Georgian coastal zone of the Black Sea and inland water reservoirs in the eastern part of Georgia. During 2006-2009, 71 phages lytic to V. cholerae were collected from these aquatic environments. The phage isolation rate was varying from 8% to 15%, depending on the sampling season and site, and the abundance of host bacteria. The majority of phages specific to V. cholerae were collected from freshwater sources. The phage isolation showed seasonal character covering warm period -from April to September. Based on basic characteristics of primary phage isolates (lytic spectrum, virion morphology and DNA restriction profiles) 23 V. cholerae -specific phages were selected for series of consecutive screenings. Comparatively wide spectrum of lytic activity was revealed in case of 14 phages specific to V. cholerae O1, and one phage - VchBS3, active against non-O1 V. cholerae. Three phages active against V. cholerae non-O1 and six V. cholerae O1 -specific phages have been studied in detail for a number of biological features (stability in different solutions, temperature-, pH- and UV- sensitivity, influence of high ionic strength etc.), considered to be additional important characteristics for selection of phages with therapeutic potential.

  18. 苏州市部分水产品分离的霍乱弧菌耐药状况及毒力基因分析%Analysis of antibiotc resistance and virulence genes of vibrio cholerae isolated from part of aquatic products in Suzhou

    Institute of Scientific and Technical Information of China (English)

    张梦寒; 王丽; 李建

    2013-01-01

    目的:分析苏州市部分养殖水产品霍乱弧菌分布情况,并对其进行耐药性分析和毒力基因检测.方法:对甲鱼、牛蛙及其养殖用水等样品,进行霍乱弧菌的分离、鉴定及耐药性分析和4种毒力基因的PCR检测.结果:分离到的18株霍乱弧菌标本中O139群8株,O1群4株,其余6株为非O1非O139群霍乱弧菌.经耐药性分析,对试验的12种抗生素均无100%敏感株;对多粘菌素B和红霉素的中介率高达83.3%.检测的4个毒力基因中,阳性率最高的是Zot基因.结论:分离到的18株霍乱弧菌血清型有一定的种属相关特性,中介耐药性突出,毒力基因阳性率较高.%Objective: To analyze the distribution of vibrio cholerae in some breeding aquatic products in Suzhou, to understand the antibiotic resistance and virulence genes. Methods; Vibrio cholerae was isolated from samples of soft - shelled turtles, bullfrogs and breeding water for identification. The detected vibrio cholerae strains were analyzed for antibiotic resistance. PCR method was adopted to detect 4 kinds of virulence genes. Results; Eighteen strains of vibrio cholerae were isolated, including serogroup 0139(8 strains) , serogroup 01 (4 strains) , non - 01 and non - 0139(6 strains). The antibiotic resistance result indicated that none of the 18 vibrio cholerae strains were 100% sensitive to 12 kinds of tested antibiotics; while the antibiotic intermediate resistance rate to polymyxin B and erythromycin was up to 83. 3%. Of four virulence genes, zonula occludens toxin gene was the most prevalent. Conclusion; There was certain species -dependent characteristics in 18 strains of vibrio cholerae by serological typing, prominent antibiotic intermediary resistance and high positive rate of virulence gene were also the features of the 18 vibrio cholerae strains.

  19. Avaliação de desinfetantes químicos de uso doméstico contra Vibrio cholerae EL TOR (amostra não toxigênica Evaluation of the effect of chemical domestic disinfectants on Vibrio cholerae EL TOR (non toxigenic strain

    Directory of Open Access Journals (Sweden)

    Jorge Timenetsky

    1992-10-01

    Brazil for microbiological qualification of chemical disinfectants for commercial purposes. Domestic disinfectants are tested in this way against Salmonella choleraesuis and Staphylococcus aureus ATCC strains, was chosen for this evaluation Vibrio cholerae in view of its current importance in Brazil, in terms of Public Health associated with the study of the disinfectant's antimicrobial activities. Nineteen disinfectant products for domestic use for available to the public were evaluated microbiologically by means of simplified Use-Dilution test with 10 carriers. The active compounds of the products included formaldeyde, phenols, cresols, quaternary ammonium compouds, chlorine and ethanol. Seven were mixtures of these. According to the recommendations for their use, sixteen products should be used undiluted. Under these conditions, 9 disinfectants were vibriocides and 7 did not demonstrate this antibacterial activity. Four products in dilutions not clearly specificated were also ineffective. The vibriocide products which must used without dilution were tested again, diluted at 1:2. These solutions did not inactivate V. cholerae showing that, microbiologically, their active compounds are used in limited concentrations. Commercial alcohol (95.5°GL at 1:3, chlorine 2.8% Água sanitária at 1:200 and Lysoform at 1:20 came up to the standards required by the test.

  20. Immunization with cholera toxin B subunit induces high-level protection in the suckling mouse model of cholera.

    Directory of Open Access Journals (Sweden)

    Gregory A Price

    Full Text Available Cholera toxin (CT is the primary virulence factor responsible for severe cholera. Vibrio cholerae strains unable to produce CT show severe attenuation of virulence in animals and humans. The pentameric B subunit of CT (CTB contains the immunodominant epitopes recognized by antibodies that neutralize CT. Although CTB is a potent immunogen and a promising protective vaccine antigen in animal models, immunization of humans with detoxified CT failed to protect against cholera. We recently demonstrated however that pups reared from mice immunized intraperitoneally (IP with 3 doses of recombinant CTB were well protected against a highly lethal challenge dose of V. cholerae N16961. The present study investigated how the route and number of immunizations with CTB could influence protective efficacy in the suckling mouse model of cholera. To this end female mice were immunized with CTB intranasally (IN, IP, and subcutaneously (SC. Serum and fecal extracts were analyzed for anti-CTB antibodies by quantitative ELISA, and pups born to immunized mothers were challenged orogastrically with a lethal dose of V. cholerae. Pups from all immunized groups were highly protected from death by 48 hours (64-100% survival. Cox regression showed that percent body weight loss at 24 hours predicted death by 48 hours, but we were unable to validate a specific amount of weight loss as a surrogate marker for protection. Although CTB was highly protective in all regimens, three parenteral immunizations showed trends toward higher survival and less weight loss at 24 hours post infection. These results demonstrate that immunization with CTB by any of several routes and dosing regimens can provide protection against live V. cholerae challenge in the suckling mouse model of cholera. Our data extend the results of previous studies and provide additional support for the inclusion of CTB in the development of a subunit vaccine against V. cholerae.

  1. Immunization with cholera toxin B subunit induces high-level protection in the suckling mouse model of cholera.

    Science.gov (United States)

    Price, Gregory A; McFann, Kim; Holmes, Randall K

    2013-01-01

    Cholera toxin (CT) is the primary virulence factor responsible for severe cholera. Vibrio cholerae strains unable to produce CT show severe attenuation of virulence in animals and humans. The pentameric B subunit of CT (CTB) contains the immunodominant epitopes recognized by antibodies that neutralize CT. Although CTB is a potent immunogen and a promising protective vaccine antigen in animal models, immunization of humans with detoxified CT failed to protect against cholera. We recently demonstrated however that pups reared from mice immunized intraperitoneally (IP) with 3 doses of recombinant CTB were well protected against a highly lethal challenge dose of V. cholerae N16961. The present study investigated how the route and number of immunizations with CTB could influence protective efficacy in the suckling mouse model of cholera. To this end female mice were immunized with CTB intranasally (IN), IP, and subcutaneously (SC). Serum and fecal extracts were analyzed for anti-CTB antibodies by quantitative ELISA, and pups born to immunized mothers were challenged orogastrically with a lethal dose of V. cholerae. Pups from all immunized groups were highly protected from death by 48 hours (64-100% survival). Cox regression showed that percent body weight loss at 24 hours predicted death by 48 hours, but we were unable to validate a specific amount of weight loss as a surrogate marker for protection. Although CTB was highly protective in all regimens, three parenteral immunizations showed trends toward higher survival and less weight loss at 24 hours post infection. These results demonstrate that immunization with CTB by any of several routes and dosing regimens can provide protection against live V. cholerae challenge in the suckling mouse model of cholera. Our data extend the results of previous studies and provide additional support for the inclusion of CTB in the development of a subunit vaccine against V. cholerae.

  2. O1群霍乱弧菌与人类疾病

    Institute of Scientific and Technical Information of China (English)

    王树坤; 李万林

    2001-01-01

    O1群霍乱弧菌(vibrio choleraer non-O1,简称非01菌)能引起人类散发腹泻已获得世界认同,但仅O1群霍乱弧菌(vibrio choleraer non-O1,简称01菌)能引起霍乱流行的看法一直沿袭到1992年,这年在印度和孟加拉国出现了由非O1菌的O139血清群霍乱弧菌(vibrio choleraer O139)引起的一次流行。

  3. Cholera Fact Sheet

    Science.gov (United States)

    ... facilities (chlorination) interventions at the household level (water filtration, chemical or solar disinfection of water, safe water ... spread of cholera and contributes to increasing antimicrobial resistance. Rapid access to treatment is essential during a ...

  4. The complete genomic analysis of an imported Vibrio cholerae from Myanmar in southwest China.

    Science.gov (United States)

    Liao, Feng; Pang, Bo; Fu, Xiaoqing; Xu, Wen; Kan, Biao; Jing, Huaiqi; Gu, Wenpeng

    2016-10-01

    We sequenced and analyzed an imported Vibrio cholerae from Mynamar in 2011 by using whole genome sequencing method in Yunnan Province, southwest China. Other 3 isolates of V. cholerae in Yunnan were also sequenced for comparing purpose. Illumina Hiseq2500 was used and the sequencing results were assembled and annotated. The comparative genomic analysis was also performed with 101 reference strains from China and abroad. The results showed the imported V. cholerae (YN2011004) had two chromosomes and one plasmid; chr1 contained 2727 predicted genes, and 958 genes for chr2. Phylogenomic tree results showed YN2011004 belonged to the seventh pandemic strain, clustered into wave 3 and clade 3B. The strain had the highly homology with SN083 and 4remapscaff isolated in 2010 from other parts of China, and clustered with SN117, VC50 remapscaff, VC57 remapscaff and SN034. These references V. cholerae mostly isolated from coastal areas of China in 2008. For the other 3 strains' comparison, it suggested that V. cholerae in 1990s in Yunnan had the close relationship with the prevalence of cholera in Southeast Asia. Therefore, we thought that the cholera in Yunnan was consistent with the epidemic trend of China, being the "sink" for external source and also acted as a "source" for spread. Moreover, we considered that the imported V. cholerae from Myanmar in 2011 actually was the exported strain from China, and it provided us a new sight for the bacterial change and evolution.

  5. Imported cholera with acute renal failure after a short business-trip to the Philippines, Germany, October 2015.

    Science.gov (United States)

    Slesak, Günther; Fleck, Ralf; Jacob, Daniela; Grunow, Roland; Schäfer, Johannes

    2016-01-01

    A German businessman developed acute watery diarrhoea after a three-day trip to the Philippines. He was admitted with severe hypotension and acute renal failure, but recovered with rapid rehydration. Vibrio cholerae O1 serotype Ogawa was isolated. Physicians need to be aware of endemic cholera in Asia including the Philippines and consider this in their pre-travel advice.

  6. Characterization of Vibrio cholerae from 1986 to 2012 in Yunnan Province, southwest China bordering Myanmar.

    Science.gov (United States)

    Gu, Wenpeng; Yin, Jianwen; Yang, Jianbin; Li, Chaoqun; Chen, Yujuan; Yin, Jie; Xu, Wen; Zhao, Shiwen; Liang, Junrong; Jing, Huaiqi; Fu, Xiaoqing

    2014-01-01

    Vibrio cholerae is an important infectious pathogen causing serious human diarrhea. We analyzed 568 V. cholerae strains isolated from 1986 to 2012 in Yunnan province, southwest China bordering Myanmar. Polymerase chain reactions for detecting virulence genes, antibiotic susceptibility tests and pulse-field gel electrophoresis (PFGE) were performed. The results showed all the strains were El Tor biotype from 1986. The ctxB subunit sequence analysis for all strains have shown that cholera between 1986 and 1995 was associated with mixed infections with El Tor and El Tor variants, while infections after 1996 were all caused by El Tor variant strains. All of the strains were sensitive to aminoglycosides and quinolone antibiotics while resistant to β-lactamase and carbapenem antibiotics increased gradually. 568 V. cholerae were divided into 218 PFGE-NotI patterns, and the isolates before 2001 and after 2011 were separated into two groups according to PFGE results. The strains isolated before 2001 were mainly referred to native cholera in Yunnan, and after 2011 were primarily referred to as imported strains from Myanmar, which showed the variation of V. cholerae in this area. The molecular characteristics of V. cholerae indicated regularity in bacterial variation and evolution in Yunnan province.

  7. Hurricanes, climate change and the cholera epidemic in Puerto Rico of 1855-1856.

    Science.gov (United States)

    Christenson, Bernard

    2008-01-01

    Hurricanes and global climate changes may affect the environmental factors of cholera dynamics in warm coastal areas, vulnerable to seasonal or sporadic outbreaks. The cholera epidemic of Puerto Rico in 1855-1856 had a profound effect on the Puerto Rican society; but it was not influenced by any climatic events, such as preceding hurricanes or storms based on past documentary sources. Particularly, the environmental non-toxigenic strains of Vibrio Cholerae in Puerto Rican water sources can maintain their pathogenic potential for sporadic or erratic toxigenic cholera outbreaks--if a "perfect storm" ever occurs.

  8. Construction and evaluation of V. cholerae O139 mutant, VCUSM21P, as a safe live attenuated cholera vaccine.

    Science.gov (United States)

    Murugaiah, Chandrika; Nik Mohd Noor, Nik Zuraina; Mustafa, Shyamoli; Manickam, Ravichandran; Pattabhiraman, Lalitha

    2014-01-01

    Cholera is a major infectious disease, affecting millions of lives annually. In endemic areas, implementation of vaccination strategy against cholera is vital. As the use of safer live vaccine that can induce protective immunity against Vibrio cholerae O139 infection is a promising approach for immunization, we have designed VCUSM21P, an oral cholera vaccine candidate, which has ctxA that encodes A subunit of ctx and mutated rtxA/C, ace and zot mutations. VCUSM21P was found not to disassemble the actin of HEp2 cells. It colonized the mice intestine approximately 1 log lower than that of the Wild Type (WT) strain obtained from Hospital Universiti Sains Malaysia. In the ileal loop assay, unlike WT challenge, 1×10⁶ and 1×10⁸ colony forming unit (CFU) of VCUSM21P was not reactogenic in non-immunized rabbits. Whereas, the reactogenicity caused by the WT in rabbits immunized with 1×10¹⁰ CFU of VCUSM21P was found to be reduced as evidenced by absence of fluid in loops administered with 1×10²-1×10⁷ CFU of WT. Oral immunization using 1×10¹⁰ CFU of VCUSM21P induced both IgA and IgG against Cholera Toxin (CT) and O139 lipopolysaccharides (LPS). The serum vibriocidal antibody titer had a peak rise of 2560 fold on week 4. Following Removable Intestinal Tie Adult Rabbit Diarrhoea (RITARD) experiment, the non-immunized rabbits were found not to be protected against lethal challenge with 1×10⁹ CFU WT, but 100% of immunized rabbits survived the challenge. In the past eleven years, V. cholerae O139 induced cholera has not been observed. However, attenuated VCUSM21P vaccine could be used for vaccination program against potentially fatal endemic or emerging cholera caused by V. cholerae O139.

  9. In Silico Structural and Functional Annotation of Hypothetical Proteins of Vibrio cholerae O139

    Science.gov (United States)

    Islam, Md. Saiful; Shahik, Shah Md.; Sohel, Md.; Patwary, Noman I. A.

    2015-01-01

    In developing countries threat of cholera is a significant health concern whenever water purification and sewage disposal systems are inadequate. Vibrio cholerae is one of the responsible bacteria involved in cholera disease. The complete genome sequence of V. cholerae deciphers the presence of various genes and hypothetical proteins whose function are not yet understood. Hence analyzing and annotating the structure and function of hypothetical proteins is important for understanding the V. cholerae. V. cholerae O139 is the most common and pathogenic bacterial strain among various V. cholerae strains. In this study sequence of six hypothetical proteins of V. cholerae O139 has been annotated from NCBI. Various computational tools and databases have been used to determine domain family, protein-protein interaction, solubility of protein, ligand binding sites etc. The three dimensional structure of two proteins were modeled and their ligand binding sites were identified. We have found domains and families of only one protein. The analysis revealed that these proteins might have antibiotic resistance activity, DNA breaking-rejoining activity, integrase enzyme activity, restriction endonuclease, etc. Structural prediction of these proteins and detection of binding sites from this study would indicate a potential target aiding docking studies for therapeutic designing against cholera. PMID:26175663

  10. 3株与O139群霍乱弧菌血清交叉凝集的麦氏弧菌的分离鉴定%Isolation and identification of 3 strains of vibrio metschnikovii isolates which cross-agglutinated with Vibrio Cholerae O139

    Institute of Scientific and Technical Information of China (English)

    李映霞; 许少洪; 黄芳; 吴琪; 曾雅

    2013-01-01

    Objective To detect and identify 3 suspected vibrio cholerae O139 isolates which isolated from 2011 to 2012.Methods The samples were isolated and cultured,serological tested and biochemical identified according to inspection standard.Results The 3 strains were identified as vibrio metschnikovii,and the 3 strains were cross-reacted with vibrio cholerae O139 diagnostic serum.Conclusions The 3 strains were not the vibrio cholerae O139,but vibrio metschnikovii.In order to prevent the possibility of false-positive results,the accuracy of identification should be assured.Especially when dealing with the cholera outbreak,the correct identification of pathogenic bacteria can provide scientific basis for control and prevent epidemics.%目的 2011-2012年对3株疑似O139群霍乱弧菌进行检测与分离鉴定.方法 依据检验标准对样本进行分离培养、血清学试验、生化反应等鉴定.结果 该3株菌为麦氏弧菌,与O139群霍乱弧菌诊断血清有交叉凝集.结论 该3菌株不是O139群霍乱弧菌,而是麦氏弧菌.为避免实验结果出现假阳性,必须进行系统生化鉴定,以确保菌株鉴定的准确性.特别是在处理霍乱疫情时,病原菌的正确鉴定可为控制疫情提供科学依据.

  11. Insights into Vibrio cholerae intestinal colonization from monitoring fluorescently labeled bacteria.

    Science.gov (United States)

    Millet, Yves A; Alvarez, David; Ringgaard, Simon; von Andrian, Ulrich H; Davis, Brigid M; Waldor, Matthew K

    2014-10-01

    Vibrio cholerae, the agent of cholera, is a motile non-invasive pathogen that colonizes the small intestine (SI). Most of our knowledge of the processes required for V. cholerae intestinal colonization is derived from enumeration of wt and mutant V. cholerae recovered from orogastrically infected infant mice. There is limited knowledge of the distribution of V. cholerae within the SI, particularly its localization along the villous axis, or of the bacterial and host factors that account for this distribution. Here, using confocal and intravital two-photon microscopy to monitor the localization of fluorescently tagged V. cholerae strains, we uncovered unexpected and previously unrecognized features of V. cholerae intestinal colonization. Direct visualization of the pathogen within the intestine revealed that the majority of V. cholerae microcolonies attached to the intestinal epithelium arise from single cells, and that there are notable regiospecific aspects to V. cholerae localization and factors required for colonization. In the proximal SI, V. cholerae reside exclusively within the developing intestinal crypts, but they are not restricted to the crypts in the more distal SI. Unexpectedly, V. cholerae motility proved to be a regiospecific colonization factor that is critical for colonization of the proximal, but not the distal, SI. Furthermore, neither motility nor chemotaxis were required for proper V. cholerae distribution along the villous axis or in crypts, suggesting that yet undefined processes enable the pathogen to find its niches outside the intestinal lumen. Finally, our observations suggest that host mucins are a key factor limiting V. cholerae intestinal colonization, particularly in the proximal SI where there appears to be a more abundant mucus layer. Collectively, our findings demonstrate the potent capacity of direct pathogen visualization during infection to deepen our understanding of host pathogen interactions.

  12. Improved laboratory capacity is required to respond better to future cholera outbreaks in Papua New Guinea

    Directory of Open Access Journals (Sweden)

    Paul Horwood

    2012-05-01

    Full Text Available Cholera was first detected in Papua New Guinea in July 2009, caused by Vibrio cholerae O1 El Tor serotype Ogawa. By late 2011, 15 500 cases had been reported throughout lowland Papua New Guinea with a case fatality rate of 3.2%. The epidemic has since slowed, with only sporadic cases reported in Western Province and the Autonomous Region of Bougainville (ARB. Accurate and timely diagnosis is a critical element of the public health response to cholera, yet in low-income countries where the burden of cholera is the greatest, diagnostic services are often limited. Here we report on the diagnostic challenges and the logistical factors that impacted on diagnosis during the first reported outbreak of cholera in Papua New Guinea.

  13. Activation of cholera toxin production by anaerobic respiration of trimethylamine N-oxide in Vibrio cholerae.

    Science.gov (United States)

    Lee, Kang-Mu; Park, Yongjin; Bari, Wasimul; Yoon, Mi Young; Go, Junhyeok; Kim, Sang Cheol; Lee, Hyung-Il; Yoon, Sang Sun

    2012-11-16

    Vibrio cholerae is a gram-negative bacterium that causes cholera. Although the pathogenesis caused by this deadly pathogen takes place in the intestine, commonly thought to be anaerobic, anaerobiosis-induced virulence regulations are not fully elucidated. Anerobic growth of the V. cholerae strain, N16961, was promoted when trimethylamine N-oxide (TMAO) was used as an alternative electron acceptor. Strikingly, cholera toxin (CT) production was markedly induced during anaerobic TMAO respiration. N16961 mutants unable to metabolize TMAO were incapable of producing CT, suggesting a mechanistic link between anaerobic TMAO respiration and CT production. TMAO reductase is transported to the periplasm via the twin arginine transport (TAT) system. A similar defect in both anaerobic TMAO respiration and CT production was also observed in a N16961 TAT mutant. In contrast, the abilities to grow on TMAO and to produce CT were not affected in a mutant of the general secretion pathway. This suggests that V. cholerae may utilize the TAT system to secrete CT during TMAO respiration. During anaerobic growth with TMAO, N16961 cells exhibit green fluorescence when stained with 2',7'-dichlorofluorescein diacetate, a specific dye for reactive oxygen species (ROS). Furthermore, CT production was decreased in the presence of an ROS scavenger suggesting a positive role of ROS in regulating CT production. When TMAO was co-administered to infant mice infected with N16961, the mice exhibited more severe pathogenic symptoms. Together, our results reveal a novel anaerobic growth condition that stimulates V. cholerae to produce its major virulence factor.

  14. Constitutive type VI secretion system expression gives Vibrio cholerae intra- and interspecific competitive advantages.

    Science.gov (United States)

    Unterweger, Daniel; Kitaoka, Maya; Miyata, Sarah T; Bachmann, Verena; Brooks, Teresa M; Moloney, Jessica; Sosa, Oscar; Silva, David; Duran-Gonzalez, Jorge; Provenzano, Daniele; Pukatzki, Stefan

    2012-01-01

    The type VI secretion system (T6SS) mediates protein translocation across the cell membrane of Gram-negative bacteria, including Vibrio cholerae - the causative agent of cholera. All V. cholerae strains examined to date harbor gene clusters encoding a T6SS. Structural similarity and sequence homology between components of the T6SS and the T4 bacteriophage cell-puncturing device suggest that the T6SS functions as a contractile molecular syringe to inject effector molecules into prokaryotic and eukaryotic target cells. Regulation of the T6SS is critical. A subset of V. cholerae strains, including the clinical O37 serogroup strain V52, express T6SS constitutively. In contrast, pandemic strains impose tight control that can be genetically disrupted: mutations in the quorum sensing gene luxO and the newly described regulator gene tsrA lead to constitutive T6SS expression in the El Tor strain C6706. In this report, we examined environmental V. cholerae isolates from the Rio Grande with regard to T6SS regulation. Rough V. cholerae lacking O-antigen carried a nonsense mutation in the gene encoding the global T6SS regulator VasH and did not display virulent behavior towards Escherichia coli and other environmental bacteria. In contrast, smooth V. cholerae strains engaged constitutively in type VI-mediated secretion and displayed virulence towards prokaryotes (E. coli and other environmental bacteria) and a eukaryote (the social amoeba Dictyostelium discoideum). Furthermore, smooth V. cholerae strains were able to outcompete each other in a T6SS-dependent manner. The work presented here suggests that constitutive T6SS expression provides V. cholerae with an advantage in intraspecific and interspecific competition.

  15. Constitutive type VI secretion system expression gives Vibrio cholerae intra- and interspecific competitive advantages.

    Directory of Open Access Journals (Sweden)

    Daniel Unterweger

    Full Text Available The type VI secretion system (T6SS mediates protein translocation across the cell membrane of Gram-negative bacteria, including Vibrio cholerae - the causative agent of cholera. All V. cholerae strains examined to date harbor gene clusters encoding a T6SS. Structural similarity and sequence homology between components of the T6SS and the T4 bacteriophage cell-puncturing device suggest that the T6SS functions as a contractile molecular syringe to inject effector molecules into prokaryotic and eukaryotic target cells. Regulation of the T6SS is critical. A subset of V. cholerae strains, including the clinical O37 serogroup strain V52, express T6SS constitutively. In contrast, pandemic strains impose tight control that can be genetically disrupted: mutations in the quorum sensing gene luxO and the newly described regulator gene tsrA lead to constitutive T6SS expression in the El Tor strain C6706. In this report, we examined environmental V. cholerae isolates from the Rio Grande with regard to T6SS regulation. Rough V. cholerae lacking O-antigen carried a nonsense mutation in the gene encoding the global T6SS regulator VasH and did not display virulent behavior towards Escherichia coli and other environmental bacteria. In contrast, smooth V. cholerae strains engaged constitutively in type VI-mediated secretion and displayed virulence towards prokaryotes (E. coli and other environmental bacteria and a eukaryote (the social amoeba Dictyostelium discoideum. Furthermore, smooth V. cholerae strains were able to outcompete each other in a T6SS-dependent manner. The work presented here suggests that constitutive T6SS expression provides V. cholerae with an advantage in intraspecific and interspecific competition.

  16. Enumeration of viable non-culturable Vibrio cholerae using propidium monoazide combined with quantitative PCR.

    Science.gov (United States)

    Wu, Bin; Liang, Weili; Kan, Biao

    2015-08-01

    The well-known human pathogenic bacterium, Vibrio cholerae, can enter a physiologically viable but non-culturable (VBNC) state under stress conditions. The differentiation of VBNC cells and nonviable cells is essential for both disease prevention and basic research. Among all the methods for detecting viability, propidium monoazide (PMA) combined with real-time PCR is popular because of its specificity, sensitivity, and speed. However, the effect of PMA treatment is not consistent and varies among different species and conditions. In this study, with an initial cell concentration of 1×10(8) CFU/ml, time and dose-effect relationships of different PMA treatments were evaluated via quantitative real-time PCR using live cell suspensions, dead cell suspensions and VBNC cell suspensions of V. cholerae O1 El Tor strain C6706. The results suggested that a PMA treatment of 20 μM PMA for 20 min was optimal under our conditions. This treatment maximized the suppression of the PCR signal from membrane-compromised dead cells but had little effect on the signal from membrane-intact live cells. In addition to the characteristics of PMA treatment itself, the initial concentration of the targeted bacteria showed a significant negative influence on the stability of PMA-PCR assay in this study. We developed a strategy that mimicked a 1×10(8) CFU/ml cell concentration with dead bacteria of a different bacterial species, the DNA of which cannot be amplified using the real time PCR primers. With this strategy, our optimal approach successfully overcame the impact of low cell density and generated stable and reliable results for counting viable cells of V. cholerae in the VBNC state.

  17. Growth Phase, Oxygen, Temperature and Starvation Affect the Development of Viable but Non-Culturable State of Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Bin eWu

    2016-03-01

    Full Text Available AbstractVibrio cholerae can enter into a viable but non-culturable (VBNC state in order to survive in unfavourable environments. In this study, we studied the roles of five physicochemical and microbiological factors or states, namely, different strains, growth phases, oxygen, temperature, and starvation, on the development of VBNC of V. cholerae in artificial sea water (ASW. Different strains of the organism, the growth phase, and oxygen levels affected the progress of VBNC development. It was found that the VBNC state was induced faster in V. cholerae serogroup O1 classical biotype strain O395 than in O1 El Tor biotype strains C6706 and N16961. When cells in different growth phases were used for VBNC induction, stationary-phase cells lost their culturability more quickly than exponential-phase cells, while induction of a totally non-culturable state took longer to achieve for stationary-phase cells in all three strains, suggesting that heterogeneity of cells should be considered. Aeration strongly accelerated the loss of culturability. During the development of the VBNC state, the culturable cell count under aeration conditions was almost 106-fold lower than under oxygen-limited conditions for all three strains. The other two factors, temperature and nutrients-rich environment, may prevent the induction of VBNC cells. At 22°C or 37°C in ASW, most of the cells rapidly died and the culturable cell count reduced from about 108 CFU/mL to 106–105 CFU/mL. The total cell counts showed that cells that lost viability were decomposed, and the viable cell counts were the same as culturable cell counts, indicating that the cells did not reach the VBNC state. VBNC state development was blocked when ASW was supplied with Luria-Bertani broth (LB, but it was not affected in ASW with M9, suggesting that specific nutrients in LB may prevent the development of VBNC state. These results revealed that the five factors evaluated in this study had different

  18. Cholera Illness and Symptoms

    Science.gov (United States)

    ... are typically no long term consequences. Persons with cholera do not become carriers of the disease after they recover, but can be reinfected if ... Diseases (NCEZID) Division of Foodborne, Waterborne, and Environmental Diseases (DFWED) ... of Health & Human Services HHS/Open USA.gov Top

  19. Cholera Transmission in Ouest Department of Haiti: Dynamic Modeling and the Future of the Epidemic

    Science.gov (United States)

    Kirpich, Alexander; Weppelmann, Thomas A.; Yang, Yang; Ali, Afsar; Morris, J. Glenn; Longini, Ira M.

    2015-01-01

    In the current study, a comprehensive, data driven, mathematical model for cholera transmission in Haiti is presented. Along with the inclusion of short cycle human-to-human transmission and long cycle human-to-environment and environment-to-human transmission, this novel dynamic model incorporates both the reported cholera incidence and remote sensing data from the Ouest Department of Haiti between 2010 to 2014. The model has separate compartments for infectious individuals that include different levels of infectivity to reflect the distribution of symptomatic and asymptomatic cases in the population. The environmental compartment, which serves as a source of exposure to toxigenic V. cholerae, is also modeled separately based on the biology of causative bacterium, the shedding of V. cholerae O1 by humans into the environment, as well as the effects of precipitation and water temperature on the concentration and survival of V. cholerae in aquatic reservoirs. Although the number of reported cholera cases has declined compared to the initial outbreak in 2010, the increase in the number of susceptible population members and the presence of toxigenic V. cholerae in the environment estimated by the model indicate that without further improvements to drinking water and sanitation infrastructures, intermittent cholera outbreaks are likely to continue in Haiti. PMID:26488620

  20. Excision dynamics of Vibrio pathogenicity island-2 from Vibrio cholerae: role of a recombination directionality factor VefA

    Directory of Open Access Journals (Sweden)

    Napolitano Michael G

    2010-11-01

    Full Text Available Abstract Background Vibrio Pathogenicity Island-2 (VPI-2 is a 57 kb region present in choleragenic V. cholerae isolates that is required for growth on sialic acid as a sole carbon source. V. cholerae non-O1/O139 pathogenic strains also contain VPI-2, which in addition to sialic acid catabolism genes also encodes a type 3 secretion system in these strains. VPI-2 integrates into chromosome 1 at a tRNA-serine site and encodes an integrase intV2 (VC1758 that belongs to the tyrosine recombinase family. IntV2 is required for VPI-2 excision from chromosome 1, which occurs at very low levels, and formation of a non-replicative circular intermediate. Results We determined the conditions and the factors that affect excision of VPI-2 in V. cholerae N16961. We demonstrate that excision from chromosome 1 is induced at low temperature and after sublethal UV-light irradiation treatment. In addition, after UV-light irradiation compared to untreated cells, cells showed increased expression of three genes, intV2 (VC1758, and two putative recombination directionality factors (RDFs, vefA (VC1785 and vefB (VC1809 encoded within VPI-2. We demonstrate that along with IntV2, the RDF VefA is essential for excision. We constructed a knockout mutant of vefA in V. cholerae N16961, and found that no excision of VPI-2 occurred, indicating that a functional vefA gene is required for excision. Deletion of the second RDF encoded by vefB did not result in a loss of excision. Among Vibrio species in the genome database, we identified 27 putative RDFs within regions that also encoded IntV2 homologues. Within each species the RDFs and their cognate IntV2 proteins were associated with different island regions suggesting that this pairing is widespread. Conclusions We demonstrate that excision of VPI-2 is induced under some environmental stress conditions and we show for the first time that an RDF encoded within a pathogenicity island in V. cholerae is required for excision of the

  1. Vibrio metoecus sp. nov., a close relative of Vibrio cholerae isolated from coastal brackish ponds and clinical specimens.

    Science.gov (United States)

    Kirchberger, Paul C; Turnsek, Maryann; Hunt, Dana E; Haley, Bradd J; Colwell, Rita R; Polz, Martin F; Tarr, Cheryl L; Boucher, Yan

    2014-09-01

    A Gram-staining-negative, curved-rod-shaped bacterium with close resemblance to Vibrio cholerae, the aetiological agent of cholera, was isolated over the course of several years from coastal brackish water (17 strains) and from clinical cases (two strains) in the United States. 16S rRNA gene identity with V. cholerae exceeded 98 % yet an average nucleotide identity based on genome data of around 86 % and multi locus sequence analysis of six housekeeping genes (mdh, adk, gyrB, recA, pgi and rpoB) clearly delineated these isolates as a distinct genotypic cluster within the V. cholerae-V. mimicus clade. Most standard identification techniques do not differentiate this cluster of isolates from V. cholerae. Only amplification of the ompW gene using V. cholerae-specific primers and a negative Voges-Proskauer test showed a difference between the two clusters. Additionally, all isolated strains differed phenotypically from V. cholerae in their ability to utilize N-acetyl-d-galactosamine and d-glucuronic acid as sole carbon sources. Furthermore, they were generally unable to infect the slime mould Dictyostelium discoideum, a widespread ability in V. cholerae. Based on these clear phenotypic differences that are not necessarily apparent in standard tests as well as average nucleotide identity and phylogeny of protein-coding genes, we propose the existence of a novel species, Vibrio metoecus sp. nov. with the type strain OP3H(T) ( = LMG 27764(T) = CIP 110643(T)). Due to its close resemblance to V. cholerae and the increasing number of strains isolated over the past several years, we suggest that V. metoecus sp. nov. is a relatively common species of the genus Vibrio, isolates of which have been identified as atypical isolates of V. cholerae in the past. Its isolation from clinical samples also indicates that strains of this species, like V. cholerae, are opportunistic pathogens.

  2. Evidence of a dominant lineage of Vibrio cholerae-specific lytic bacteriophages shed by cholera patients over a 10-year period in Dhaka, Bangladesh.

    Science.gov (United States)

    Seed, Kimberley D; Bodi, Kip L; Kropinski, Andrew M; Ackermann, Hans-Wolfgang; Calderwood, Stephen B; Qadri, Firdausi; Camilli, Andrew

    2011-01-01

    Lytic bacteriophages are hypothesized to contribute to the seasonality and duration of cholera epidemics in Bangladesh. However, the bacteriophages contributing to this phenomenon have yet to be characterized at a molecular genetic level. In this study, we isolated and sequenced the genomes of 15 bacteriophages from stool samples from cholera patients spanning a 10-year surveillance period in Dhaka, Bangladesh. Our results indicate that a single novel bacteriophage type, designated ICP1 (for the International Centre for Diarrhoeal Disease Research, Bangladesh cholera phage 1) is present in all stool samples from cholera patients, while two other bacteriophage types, one novel (ICP2) and one T7-like (ICP3), are transient. ICP1 is a member of the Myoviridae family and has a 126-kilobase genome comprising 230 open reading frames. Comparative sequence analysis of ICP1 and related isolates from this time period indicates a high level of genetic conservation. The ubiquitous presence of ICP1 in cholera patients and the finding that the O1 antigen of lipopolysaccharide (LPS) serves as the ICP1 receptor suggest that ICP1 is extremely well adapted to predation of human-pathogenic V. cholerae O1.

  3. 疑似霍乱弧菌鉴定及分子分型检测%Characterization and molecular typing of suspected Vibrio cholerae isolates

    Institute of Scientific and Technical Information of China (English)

    陈琛; 王勇; 黄留玉; 李承毅; 宋宏彬; 王中强; 柳楠; 刘雯静; 李婧; 薛文仲; 张伶; 杜昕颖; 邱少富

    2011-01-01

    Objective To identify two suspected Vibrio cholerae isolates from patients with intestinal diseases diagnosed in a hospital in Beijing city and to detect their main virulence genes and analyze their genotypes. Methods The suspected V. cholerae isolates were identified with API 20E strips, serotype and 16S rRNA analysis. PCR was used to detect the distribution of the eight major virulence genes, and the genetic association was measured with pulsed-field gel electrophoresis (PFGE). Results The two suspected V. cholerae isolates were confirmed as non-O1/non-O139 V. cholerae. 16S rRNA analysis showed that the two isolates was 100% homologous to those from NCBI database. Virulence genes hylA ,ompW and toxR were detected. The two strains showed different banding patterns with PFGE. Conclusion The two suspected strains were identified to be non-O1/non-O139 V. cholera and the pathogenic factors may be related to the virulence genes hylA,ompW and toxR. The two strains were distant in evolutionary relationship. The result indicates that different types of non-O1/non-O139 V. cholera exist in the city.%目的 对北京市某医院肠道门诊2株疑似霍乱弧菌进行鉴定,检测其主要毒力基因,并分析其基因型别特征.方法 利用细菌生化鉴定条(API 20 E)进行生化鉴定,玻片凝集法确定其血清型别,应用PCR及测序技术分析其16 S rRNA基因;PCR检测其8个主要毒力基因,脉冲场凝胶电泳(PFGE)分析其基因型别.结果 经鉴定,该2株疑似霍乱菌株均为非O 1非139群霍乱弧菌,经16 S rRNA分析与美国国家生物技术信息中心数据库中霍乱弧菌相似性达100%,均包含毒力基因hylA、ompW和toxR,PFGE结果显示2株菌具有完全不同的带型.结论 2株疑似霍乱弧菌为非O 1非139群霍乱弧菌,其致病因素与(hylA)、(ompW)、(toxR)毒力基因有关,并且2株菌遗传关系较远.

  4. Invariant recognition of polychromatic images of Vibrio cholerae 01

    Science.gov (United States)

    Alvarez-Borrego, Josue; Mourino-Perez, Rosa R.; Cristobal, Gabriel; Pech-Pacheco, Jose L.

    2002-04-01

    Cholera is an acute intestinal infectious disease. It has claimed many lives throughout history, and it continues to be a global health threat. Cholera is considered one of the most important emergence diseases due its relation with global climate changes. Automated methods such as optical systems represent a new trend to make more accurate measurements of the presence and quantity of this microorganism in its natural environment. Automatic systems eliminate observer bias and reduce the analysis time. We evaluate the utility of coherent optical systems with invariant correlation for the recognition of Vibrio cholerae O1. Images of scenes are recorded with a CCD camera and decomposed in three RGB channels. A numeric simulation is developed to identify the bacteria in the different samples through an invariant correlation technique. There is no variation when we repeat the correlation and the variation between images correlation is minimum. The position-, scale-, and rotation-invariant recognition is made with a scale transform through the Mellin transform. The algorithm to recognize Vibrio cholerae O1 is the presence of correlation peaks in the green channel output and their absence in red and blue channels. The discrimination criterion is the presence of correlation peaks in red, green, and blue channels.

  5. Characterization of Vibrio cgolerae non-O1 serogroups obtained from an outbreak of diarrhea in Lima, Peru.

    Science.gov (United States)

    Dalsgaard, A; Albert, M J; Taylor, D N; Shimada, T; Meza, R; Serichantalergs, O; Echeverria, P

    1995-10-01

    In February 1994, an outbreak of diarrhea caused by non-O1 Vibrio cholerae occurred among volunteers in a vaccine trial study area in Lima, Peru. Clinically, 95% of the patients presented with liquid diarrhea with either no or mild dehydration. Serogrouping of 58 isolates recovered from diarrheal patients affected in the outbreak revealed seven different serogroups, with serogroups O10 (21%) and O12 (65%) being predominant. Most of these isolates were susceptible to a variety of antimicrobial agents. None of the 58 isolates hybridized with a DNA probe previously used to detect the gene encoding the heat-stable enterotoxin NAG-ST or produced cholera toxin as assessed by GM1 ganglioside enzyme-linked immunosorbent assay. Ribotyping exhibited 10 different BglI ribotype patterns among the 58 V. cholera non-O1 strains studied. However, ribotyping showed that all isolates belonging to serogroup O12 exhibited identical ribotypes and that 83% of the serogroup O10 isolates belonged to another identical ribotype, thus showing excellent correlation between ribotypes and serogroups. Among a group of O10 and O12 isolates selected for virulence studies, none produced enterotoxin whereas the majority produced a cytotoxin, as assessed in Y1 and HeLa cells. These isolates were also negative for the gene encoding zonula occludens toxin (Zot) as assessed by a PCR assay. The isolates tested showed strong adherence and some degree of invasion in the HEp-2 cell assay, whereas none of the isolates was positive in the PCR assay for the gene encoding the toxin coregulated pilus subunit A antigen (tcpA). In the removable intestinal tie adult rabbit diarrhea model, O10 and O12 serogroup isolates produced severe diarrhea and occasionally death when rabbits were challenged with 10(10) bacterial cells. Fluid accumulation was shown in the rabbit intestinal loop test when whole cultures were injected. No significant difference in virulence was shown between serogroup O10 and O12 isolates. This

  6. Nonredundant Roles of Iron Acquisition Systems in Vibrio cholerae.

    Science.gov (United States)

    Peng, Eric D; Wyckoff, Elizabeth E; Mey, Alexandra R; Fisher, Carolyn R; Payne, Shelley M

    2015-12-07

    Vibrio cholerae, the causative agent of the severe diarrheal disease cholera, thrives in both marine environments and the human host. To do so, it must encode the tools necessary to acquire essential nutrients, including iron, under these vastly different conditions. A number of V. cholerae iron acquisition systems have been identified; however, the precise role of each system is not fully understood. To test the roles of individual systems, we generated a series of mutants in which only one of the four systems that support iron acquisition on unsupplemented LB agar, Feo, Fbp, Vct, and Vib, remains functional. Analysis of these mutants under different growth conditions showed that these systems are not redundant. The strain carrying only the ferrous iron transporter Feo grew well at acidic, but not alkaline, pH, whereas the ferric iron transporter Fbp promoted better growth at alkaline than at acidic pH. A strain defective in all four systems (null mutant) had a severe growth defect under aerobic conditions but accumulated iron and grew as well as the wild type in the absence of oxygen, suggesting the presence of an additional, unidentified iron transporter in V. cholerae. In support of this, the null mutant was only moderately attenuated in an infant mouse model of infection. While the null mutant used heme as an iron source in vitro, we demonstrate that heme is not available to V. cholerae in the infant mouse intestine.

  7. 福建省2009~2011年霍乱监测分析%Surveillance of cholera in Fujian Province in 2009~2011

    Institute of Scientific and Technical Information of China (English)

    罗朝晨; 郑金凤; 陈爱平; 徐海滨; 杨劲松

    2012-01-01

    目的 掌握福建省霍乱流行规律和霍乱弧菌在环境水体及食品的污染情况,为指导今后的霍乱防治工作提供依据.方法 全省医疗机构肠道门诊开展腹泻病人监测,各级霍乱监测点4~11月份同时开展水体及海水产品等食品监测.结果 2009~2011年福建省各级霍乱监测点共检测腹泻病人粪便标本56922份,发现5例霍乱实验室确诊病人.2009~2011年福建省各级霍乱监测点检测水体及海水产品等食品标本62471份,共检出O1群和O139群霍乱弧菌102株,总阳性率为0.16%,其中O1群稻叶型85株,O1群小川型17株,均为非产毒株.结论 开展霍乱监测是早期发现疫情、及时掌握疫情动态的重要措施,今后应加强对牛蛙等海水产品的霍乱弧菌污染状况监测,以及加大外环境检索力度和加强各种水体的消毒措施,防止疫情扩散.%Objective To grasp the epidemic regularity and contamination conditions of Vibrio cholerae in water bodies and foods so as to provide basis for preventing and controlling cholera in Fujian. Methods Surveillances of diarrhea patients were carried out in diarrhea outpatient of the medical institutions in the whole province. Samples of food,water bodies and seawater aquatic products were collected and monitored from April to November 2009~2011. Results Five cholera cases were laboratory-confirmed from 56 922 stool specimens of diarrhea patients in all levels of monitoring sites in Fujian from 2009 to 2011. 102 strains of 01 and 0139 V. cholerae were detected from 62 471 specimens of water bodies and seawater aquatic products in all levels of monitoring sites. The total positive rate was 0.16% ,in which 85 strains were O1 Inaba,17 strains were 01 Ogawa,all were non-toxicgenic strains. Conclusion The pathogenic surveillance of cholera is the important measure for early detection of cholera outbreak and understand the variation trend of the outbreak. Surveillances of seawater aquatic

  8. (p)ppGpp, a Small Nucleotide Regulator, Directs the Metabolic Fate of Glucose in Vibrio cholerae.

    Science.gov (United States)

    Oh, Young Taek; Lee, Kang-Mu; Bari, Wasimul; Raskin, David M; Yoon, Sang Sun

    2015-05-22

    When V. cholerae encounters nutritional stress, it activates (p)ppGpp-mediated stringent response. The genes relA and relV are involved in the production of (p)ppGpp, whereas the spoT gene encodes an enzyme that hydrolyzes it. Herein, we show that the bacterial capability to produce (p)ppGpp plays an essential role in glucose metabolism. The V. cholerae mutants defective in (p)ppGpp production (i.e. ΔrelAΔrelV and ΔrelAΔrelVΔspoT mutants) lost their viability because of uncontrolled production of organic acids, when grown with extra glucose. In contrast, the ΔrelAΔspoT mutant, a (p)ppGpp overproducer strain, exhibited better growth in the presence of the same glucose concentration. An RNA sequencing analysis demonstrated that transcriptions of genes consisting of an operon for acetoin biosynthesis were markedly elevated in N16961, a seventh pandemic O1 strain, but not in its (p)ppGpp(0) mutant during glucose-stimulated growth. Transposon insertion in acetoin biosynthesis gene cluster resulted in glucose-induced loss of viability of the ΔrelAΔspoT mutant, further suggesting the crucial role of acetoin production in balanced growth under glucose-rich environments. Additional deletion of the aphA gene, encoding a negative regulator for acetoin production, failed to rescue the (p)ppGpp(0) mutant from the defective glucose-mediated growth, suggesting that (p)ppGpp-mediated acetoin production occurs independent of the presence of AphA. Overall, our results reveal that (p)ppGpp, in addition to its well known role as a stringent response mediator, positively regulates acetoin production that contributes to the successful glucose metabolism and consequently the proliferation of V. cholerae cells under a glucose-rich environment, a condition that may mimic the human intestine.

  9. Experimental study on administration of microeneapsulated vibrio cholera vaccine.

    Institute of Scientific and Technical Information of China (English)

    1998-01-01

    Objective: To study the effect of biodegradable microspheres as a vaccine delivery system for V. cholera antigen. Methods: The outer membrane protein (OMP, 41KDa) was obtained from the strain Enaba 569B, and the OMP was encapsulated in the biodegrad-

  10. From cholera to corals: Viruses as drivers of virulence in a major coral bacterial pathogen

    KAUST Repository

    Weynberg, Karen D.

    2015-12-08

    Disease is an increasing threat to reef-building corals. One of the few identified pathogens of coral disease is the bacterium Vibrio coralliilyticus. In Vibrio cholerae, infection by a bacterial virus (bacteriophage) results in the conversion of non-pathogenic strains to pathogenic strains and this can lead to cholera pandemics. Pathogenicity islands encoded in the V. cholerae genome play an important role in pathogenesis. Here we analyse five whole genome sequences of V. coralliilyticus to examine whether virulence is similarly driven by horizontally acquired elements. We demonstrate that bacteriophage genomes encoding toxin genes with homology to those found in pathogenic V. cholerae are integrated in V. coralliilyticus genomes. Virulence factors located on chromosomal pathogenicity islands also exist in some strains of V. coralliilyticus. The presence of these genetic signatures indicates virulence in V. coralliilyticus is driven by prophages and other horizontally acquired elements. Screening for pathogens of coral disease should target conserved regions in these elements.

  11. Antibiotic resistance of vibrio cholerae: special considerations of R-plasmids.

    Science.gov (United States)

    Kuwahara, S

    1978-09-01

    Studies on the transmission of R plasmid by conjugation between enterobacteria and vibrio or related bacteria were reviewed. The majority of the reports confirmed successful transmission from enterobacteria to Vibrio cholerae and related species, although the transmission frequencies were extremely low and the transmitted R plasmid was very unstable except for thermosensitive kanamycin plasmid and usual R plasmid coexisting with P plasmid. Strains of V. cholerae and Aeromonas liquefaciens as well as A. salmonicida bearing R plasmid were detected in nature. R plasmid was relatively unstable in V. cholerae strains with which transmission of R plasmid to enterobacteria was confirmed. At present, only 3 R plasmids have been obtained from naturally occurring strains of V. cholerae. Although the 2 European plasmids belong to the C incompatibility group with 98 megadalton closed covalent circular DNA molecule, one plasmid belongs to the J group with more than 25 megadalton molecular weight, and no CCC of satelite DNA was detected in bacteria harboring this plasmid.

  12. From cholera to corals: Viruses as drivers of virulence in a major coral bacterial pathogen

    Science.gov (United States)

    Weynberg, Karen D.; Voolstra, Christian R.; Neave, Matthew J.; Buerger, Patrick; van Oppen, Madeleine J. H.

    2015-01-01

    Disease is an increasing threat to reef-building corals. One of the few identified pathogens of coral disease is the bacterium Vibrio coralliilyticus. In Vibrio cholerae, infection by a bacterial virus (bacteriophage) results in the conversion of non-pathogenic strains to pathogenic strains and this can lead to cholera pandemics. Pathogenicity islands encoded in the V. cholerae genome play an important role in pathogenesis. Here we analyse five whole genome sequences of V. coralliilyticus to examine whether virulence is similarly driven by horizontally acquired elements. We demonstrate that bacteriophage genomes encoding toxin genes with homology to those found in pathogenic V. cholerae are integrated in V. coralliilyticus genomes. Virulence factors located on chromosomal pathogenicity islands also exist in some strains of V. coralliilyticus. The presence of these genetic signatures indicates virulence in V. coralliilyticus is driven by prophages and other horizontally acquired elements. Screening for pathogens of coral disease should target conserved regions in these elements. PMID:26644037

  13. The complete genome sequence of Vibrio cholerae: a tale of two chromosomes and of two lifestyles.

    Science.gov (United States)

    Schoolnik, G K; Yildiz, F H

    2000-01-01

    Vibrio cholerae O1 has figured prominently in the history of infectious diseases as a cause of periodic global epidemics, an affliction of refugees in areas of social strife and as the disease first subjected to modern epidemiological analysis during the classic investigations of John Snow in mid-19th century London [1]. Thus, publication of the entire genome sequence of V. cholerae O1 (biotype El Tor) in Nature [2] by a consortium of investigators from The Institute for Genomic Research, the University of Maryland and Harvard Medical School is properly regarded as an historic event that will trigger a paradigm shift in the study of this organism.

  14. Susceptibilidad a agentes antimicrobianos de cepas vacunales atenuadas de Vibrio cholerae

    OpenAIRE

    Angela Mariana Zayas Tamayo; Grether Barreras García; Karen Marrero Domínguez

    2014-01-01

    El cólera es una enfermedad diarreica aguda causada por la presencia del Vibrio cholerae O1 en el intestino delgado por la ingestión de alimentos o aguas contaminadas con Vibrio cholerae O1 . En particular, las vacunas atenuadas parecen ser una de las variantes más promisorias pa ra su prevención y para su aprobación, se hace necesaria su evaluación clínica. La agencia reguladora cubana [Centro para el Control Estatal de Medicamentos, Equipos y Dispositivos Medicamentos (CECMED)] tiene entre ...

  15. 霍乱弧菌及肠出血性大肠杆菌多重PCR检测方法的建立%Development of multiplex PCR for detection of Vibrio cholerae and enterohemorrhagic Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    刘彦晶; 吴大成; 孟福强; 袁洁; 孙洋; 冯书章

    2012-01-01

    A multiplex PCR method was developed for the rapid detection of genes encoding Shiga toxins 1 and 2(stx1 and stx2) of enterohemorrhagic Escherichia coli(EHEC) and Cholera toxin gene(ctx) of Vibrio cholerae.Three pairs of primers were synthesized.The size of amplified products is different,so it is easy to distinguish the bands on agarose gel.The method includes enrichment before the PCR reaction.The specificity of the multiplex PCR method was determined by using 25 strains of pure-cultured bacteria,including Vibrio cholerae O1 and O139 and 4 EHEC strains.All 6 EHEC strains and Vibrio cholerae strains produced positive bands,whereas the others did not.The sensitivity was evaluated by detection of artificially contaminated samples,with a range of 100-1 000 CFU/mL in contaminated water samples and 1 CFU/mL after enrichment.These results indicated that the Multi-PCR method exhibited high specificity and sensitivity in the detection of EHEC and Vibrio cholerae.It could be used to detec environment samples contaminated by EHEC and Vibrio cholerae.%针对霍乱弧菌肠毒素和肠出血性大肠杆菌志贺毒素基因(ctx、stx1和stx2)设计引物,扩增大小不同的特异性片段,优化反应条件,建立多重PCR方法,对人工污染的水样品进行模拟检测。结果显示,多重PCR扩增系统针对目的菌具有高度的特异性。敏感性试验证实,当多重PCR反应体系中模板含量在10CFU时仍能检出。模拟水样品多重PCR检测的敏感性试验证明,人工污染的河水直接集菌检测敏感性为100~1 000CFU/mL,增菌培养后多重PCR检测敏感性可达1CFU/mL。本试验于同一PCR体系检测霍乱弧菌和肠出血性大肠杆菌的毒素基因,可用于这2种致腹泻性病原菌的快速检测。

  16. Environmental Monitoring of Endemic Cholera

    Science.gov (United States)

    ElNemr, W.; Jutla, A. S.; Constantin de Magny, G.; Hasan, N. A.; Islam, M.; Sack, R.; Huq, A.; Hashem, F.; Colwell, R.

    2012-12-01

    Cholera remains a major public health threat. Since Vibrio cholerae, the causative agent of the disease, is autochthonous to riverine, estuarine, and coastal waters, it is unlikely the bacteria can be eradicated from its natural habitat. Prediction of disease, in conjunction with preventive vaccination can reduce the prevalence rate of a disease. Understanding the influence of environmental parameters on growth and proliferation of bacteria is an essential first step in developing prediction methods for outbreaks. Large scale geophysical variables, such as SST and coastal chlorophyll, are often associated with conditions favoring growth of V. cholerae. However, local environmental factors, meaning biological activity in ponds from where the bulk of populations in endemic regions derive water for daily usage, are either neglected or oversimplified. Using data collected from several sites in two geographically distinct locations in South Asia, we have identified critical local environmental factors associated with cholera outbreak. Of 18 environmental variables monitored for water sources in Mathbaria (a coastal site near the Bay of Bengal) and Bakergonj (an inland site) of Bangladesh, water depth and chlorophyll were found to be important factors associated with initiation of cholera outbreaks. Cholera in coastal regions appears to be related to intrusion. However, monsoonal flooding creates conditions for cholera epidemics in inland regions. This may be one of the first attempts to relate in-situ environmental observations with cholera. We anticipate that it will be useful for further development of prediction models in the resource constrained regions.

  17. Survival of Vibrio cholerae in nutrient-poor environments is associated with a novel "persister" phenotype.

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    Mohamma Jubair

    Full Text Available In response to antibiotic and/or environmental stress, some species of bacteria shift to a "persister" phenotype. Although toxigenic Vibrio cholerae, responsible for the disease cholera, can be found in nutrient-poor aquatic environments in endemic areas, the underlying mechanism(s by which culturable cells persist in these environmental reservoirs is largely unknown. Here we report that introduction of V. cholerae into a nutrient-poor filter sterilized lake water (FSLW microcosm promoted a shift to what we have defined as a "persister" phenotype (PP which was culturable for >700 days. Direct transfer of PP of V. cholerae from original microcosms to freshly prepared FSLW resulted in the same pattern of persistence seen in the original microcosms. Scanning electron microscopy of cells persisting for over 700 days demonstrated cell morphologies that were very small in size, with a high degree of aggregation associated with flagella emanating from all aspects of the cell. V. cholerae PP cells reverted to a typical V. cholerae morphology when transferred to nutrient-rich L- broth. Cell-free supernatants obtained from microcosms at 24 hours, 180 days, and 700 days all showed >2-fold increase in CAI-1 signaling molecules, consistent with quorum sensing activity, as has been described for Pseudomonas aeruginosa persister cells. Chitin and phosphate promoted cell growth. Our data suggest that nutrient stress can select a V. cholerae persister phenotype in environmental reservoirs, with these strains then seeding subsequent cholera epidemics in response to chitin and phosphate availability.

  18. Hyperinfectivity: a critical element in the ability of V. cholerae to cause epidemics?

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    David M Hartley

    2006-01-01

    Full Text Available Cholera is an ancient disease that continues to cause epidemic and pandemic disease despite ongoing efforts to limit its spread. Mathematical models provide one means of assessing the utility of various proposed interventions. However, cholera models that have been developed to date have had limitations, suggesting that there are basic elements of cholera transmission that we still do not understand.Recent laboratory findings suggest that passage of Vibrio cholerae O1 Inaba El Tor through the gastrointestinal tract results in a short-lived, hyperinfectious state of the organism that decays in a matter of hours into a state of lower infectiousness. Incorporation of this hyperinfectious state into our disease model provides a much better fit with the observed epidemic pattern of cholera. These findings help to substantiate the clinical relevance of laboratory observations regarding the hyperinfectious state, and underscore the critical importance of human-to-human versus environment-to-human transmission in the generation of epidemic and pandemic disease.To have maximal impact on limiting epidemic spread of cholera, interventions should be targeted toward minimizing risk of transmission of the short-lived, hyperinfectious form of toxigenic Vibrio cholerae. The possibility of comparable hyperinfectious states in other major epidemic diseases also needs to be evaluated and, as appropriate, incorporated into models of disease prevention.

  19. Cholera epidemic associated with raw vegetables--Lusaka, Zambia, 2003-2004.

    Science.gov (United States)

    2004-09-03

    Zambia experienced widespread cholera epidemics in 1991 (13,154 cases), 1992 (11,659), and 1999 (11,327). In response to the large outbreak in 1999, the Zambian Ministry of Health (ZMOH) urged use of in-home chlorination with the locally produced solution, Clorin, and the practice increased substantially Clorin had been introduced in Zambia in 1998 as part of the Safe Water System (SWS), a point-of-use water disinfection and safe-water storage strategy launched by the Society for Family Health, in partnership with ZMOH, the U.S. Agency for International Development, and CDC. Although no outbreaks were reported during 2000-2002, cholera remained endemic. Epidemic cholera returned to Zambia in November 2003, when cases of toxigenic Vibrio cholerae O1, serotype Ogawa, biotype El Tor were confirmed in the capital city, Lusaka. During November 28, 2003-January 4, 2004, an estimated 2,529 cholera cases and 128 cholera deaths (case-fatality rate [CFR] = 5.1%) occurred in Lusaka. In February 2004, the Lusaka District Health Management Team (LDHMT) invited CDC to assist in an investigation of the epidemic. This report summarizes the results of that investigation, which implicated foodborne transmission via raw vegetables and demonstrated a protective role for hand washing with soap. The results underscore the importance of hygiene, clean water, and sanitary food handling for cholera prevention.

  20. Genome engineering in Vibrio cholerae

    DEFF Research Database (Denmark)

    Val, Marie-Eve; Skovgaard, Ole; Ducos-Galand, Magaly

    2012-01-01

    importance in public health, Vibrio cholerae, the causative agent of cholera, has become a preferred model to study bacteria with multipartite genomes. However, most in vivo studies in V. cholerae have been hampered by its genome architecture, as it is difficult to give phenotypes to a specific chromosome....... This difficulty was surmounted using a unique and powerful strategy based on massive rearrangement of prokaryotic genomes. We developed a site-specific recombination-based engineering tool, which allows targeted, oriented, and reciprocal DNA exchanges. Using this genetic tool, we obtained a panel of V. cholerae...... in V. cholerae and the general question concerning bacteria carrying circular chromosomes--by looking at the effect of chromosome size on topological issues. In this article, we show that Dam, RctB, and ParA2/ParB2 are strictly essential for chrII origin maintenance, and we formally demonstrate...

  1. Epidemic cholera spreads like wildfire

    Science.gov (United States)

    Roy, Manojit; Zinck, Richard D.; Bouma, Menno J.; Pascual, Mercedes

    2014-01-01

    Cholera is on the rise globally, especially epidemic cholera which is characterized by intermittent and unpredictable outbreaks that punctuate periods of regional disease fade-out. These epidemic dynamics remain however poorly understood. Here we examine records for epidemic cholera over both contemporary and historical timelines, from Africa (1990-2006) and former British India (1882-1939). We find that the frequency distribution of outbreak size is fat-tailed, scaling approximately as a power-law. This pattern which shows strong parallels with wildfires is incompatible with existing cholera models developed for endemic regions, as it implies a fundamental role for stochastic transmission and local depletion of susceptible hosts. Application of a recently developed forest-fire model indicates that epidemic cholera dynamics are located above a critical phase transition and propagate in similar ways to aggressive wildfires. These findings have implications for the effectiveness of control measures and the mechanisms that ultimately limit the size of outbreaks.

  2. Surveillance for Vibrio cholerae in Wuzhou city in 2009-2014%2009-2014年梧州市霍乱弧菌监测结果

    Institute of Scientific and Technical Information of China (English)

    冼桂江; 盘珍梅; 彭美薇; 覃敏兰

    2016-01-01

    Objective To know the prevalence of Vibrio cholerae in the environment and aquatic products in Wuzhou city , and monitor the shift of its serotype and virulence distribution, and provide reference for making prevention policy. Methods A 6-year surveillance program was conducted to detect V . cholerae from aquatic products and cooked food at markets , from rivers and ponds (from May to October each year) as well as from diarrhea cases at hospitals (all year round). Results No V. cholerae was detected from 600 food samples and 7 693 patients. The positive rate was 2.9%(69/2 375)in the environmental samples and aquatic products. The majority serogroup of V. cholerae was O1 (91.3%, 63/69) and the remaining was O139 (8.7%, 6/69). The O1 strains belonged to two serotypes: Inaba (49.3%, 34/69)and Ogawa(42.0%, 29/69). All the V. cholerae isolates were non-toxigenic. Conclusion The environment and aquatic products in Wuzhou city were contaminated with V. cholerae with serogroup and serotype diversity. Continue surveillance is necessary to prevent the spread of toxigenic V. cholerae.%目的:为了解梧州市外环境霍乱弧菌的存在和水产品的污染状况,掌握其菌型、分布、毒力等动态变化情况,为霍乱防控工作提供科学依据。方法采集梧州市两城区各大集贸市场销售摊档水产品、熟食制品、市区河水和池塘水(每年5~10月监测)以及医院腹泻就诊病人(每年1~12月监测)样品,并对其进行霍乱弧菌监测。结果熟食制品(600份)、腹泻病人大便标本(7693份)均未检出霍乱弧菌。河水、池塘水和海水产品阳性率为2.9%(69/2375)。其中O1群占91.3%(63/69),O139群占8.7%(6/69);O1群血清学分型:稻叶型49.3%(34/69),小川型42.0%(29/69)。各血清型CT毒力基因均为阴性。结论梧州市外环境水体、海水产品受霍乱弧菌污染严重,且菌株型多元化,应予继

  3. Social and cultural features of cholera and shigellosis in peri-urban and rural communities of Zanzibar

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    Hutubessy Raymond

    2010-11-01

    Full Text Available Abstract Background Responding to the high burden of cholera in developing countries, the WHO now considers vaccination as a supplement to the provision of safe drinking water and improved sanitation in the strategy for cholera control in endemic settings. Cultural concepts of illness affect many aspects of public health. In the first step of a two-step strategy to examine determinants of cholera vaccine acceptance, this study identified social and cultural features of diarrhoeal illness for cholera control in endemic communities. Methods A cultural epidemiological study with locally adapted vignette-based interviews was conducted in two cholera-endemic communities of Zanzibar. A random sample of unaffected peri-urban (n = 179 and rural (n = 177 adults was interviewed to study community ideas of cholera and shigellosis, considering categories of distress, perceived causes, and help-seeking behaviour. Results Cholera was recognised by 88%. Symptoms of dehydration were most prominent in reports at the peri-urban site. Interference with work leading to strain on household finances was frequently emphasised. Dirty environment was the most prominent perceived cause, followed by unsafe drinking water and germ-carrying flies. Causes unrelated to the biomedical basis of cholera were reported more often by rural respondents. Rural women had more difficulty (20% to identify a cause than men (7.1%, p = 0.016. Peri-urban self treatment emphasised rehydration; the rural community preferred herbal treatment and antibiotics. Shigellosis was recognised by 70%. Fewer regarded it as very serious compared with cholera (76% vs. 97%, p Conclusions This study clarified local views of cholera and shigellosis relevant for diarrhoeal disease control in Zanzibar. The finding that rural women were less likely than men to specify causes of cholera suggests more attention to them is required. Better health education is needed for cholera in rural areas and for shigellosis

  4. Role of Ectoine in Vibrio cholerae Osmoadaptation

    OpenAIRE

    Pflughoeft, Kathryn J.; Kierek, Katharine; Paula I Watnick

    2003-01-01

    Vibrio cholerae is both an intestinal pathogen and a microbe in the estuarine community. To persist in the estuarine environment, V. cholerae must adjust to changes in ionic composition and osmolarity. These changes in the aquatic environment have been correlated with cholera epidemics. In this work, we study the response of V. cholerae to increases in environmental osmolarity. Optimal growth of V. cholerae in minimal medium requires supplementation with 200 mM NaCl and KCl. However, when the...

  5. Field evaluation of a rapid immunochromatographic dipstick test for the diagnosis of cholera in a high-risk population

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    Park Taesung

    2006-02-01

    Full Text Available Abstract Background Early detection of cholera outbreaks is crucial for the implementation of the most appropriate control strategies. Methods The performance of an immunochromatographic dipstick test (Institute Pasteur, Paris, France specific for Vibrio cholerae O1 was evaluated in a prospective study in Beira, Mozambique, during the 2004 cholera season (January-May. Fecal specimens were collected from 391 patients with acute watery nonbloody diarrhea and tested by dipstick and conventional culture. Results The overall sensitivity and specificity of the rapid test compared to culture were 95% (95% confidence interval [CI]: 91%–99% and 89% (95% CI: 86%–93%, respectively. After stratification by type of sample (rectal swab/bulk stool and severity of diarrhea, the sensitivity ranged between 85% and 98% and specificity between 77% and 97%. Conclusion This one-step dipstick test performed well in the diagnosis of V. cholerae O1 in a setting with seasonal outbreaks where rapid tests are most urgently needed.

  6. Virulence factors in environmental and clinical Vibrio cholerae from endemic areas in Kenya

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    Racheal W. Kimani

    2014-04-01

    Full Text Available Background: Since 1971, Kenya has had repeated cholera outbreaks. However, the cause of seasonal epidemics of cholera is not fully understood and neither are the factors that drive epidemics, both in Kenya and globally.Objectives: The objectives of the study were to determine the environmental reservoirs of V. cholerae during an interepidemic period in Kenya and to characterise their virulence factors.Methods: One hundred (50 clinical, 50 environmental samples were tested for V. cholerae isolates using both simplex and multiplex polymerase chain reaction.Results: Both sediments and algae from fishing and landing bays yielded isolates of V. cholerae. Clinical strains were characterised along with the environmental strains for comparison. All clinical strains harboured ctxA, tcpA (El Tor, ompU, zot, ace, toxR, hylA (El Tor and tcpI genes. Prevalence for virulence genes in environmental strains was hylA (El Tor (10%, toxR (24%, zot (22%, ctxA (12%,tcpI (8%, hylA (26% and tcpA (12%.Conclusion: The study sites, including landing bays and beaches, contained environmental V. cholerae, suggesting that these may be reservoirs for frequent epidemics. Improved hygiene and fish-handling techniques will be important in reducing the persistence of reservoirs.

  7. A reexamination of the O1 lipopolysaccharide antigen group of Escherichia coli.

    Science.gov (United States)

    Moll, A; Kusecek, B; Pluschke, G; Morelli, G; Kamke, M; Jann, B; Jann, K; Achtman, M

    1986-01-01

    A total of 64 Escherichia coli strains of the O1 serogroup were tested for the migration pattern of their lipopolysaccharides (LPS) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. O1:K1 and O1:K51 strains of the OMP5 outer membrane protein pattern possessed LPS with a doublet pattern (O1A1) or the lowermost band of the O1A1 doublet (O1A2). O1:K1 strains of the OMP9 pattern possessed LPS referred to as O1A, which corresponded to the uppermost band of the O1A1 doublet pattern. A few O1:K? strains possessed LPS of different migration patterns (O1B and O1C). O1A and O1A1 LPS were indistinguishable by chemical techniques, and both reacted with each of 10 different monoclonal antibodies tested. However, O1A1 had an additional epitope within the additional band in each doublet, as demonstrated by adsorption experiments with hyperimmune rabbit sera followed by Western blotting. Furthermore, purified polysaccharide from O1A bacteria was incapable of inhibition in enzyme-linked immunosorbent assays performed with O1A1 LPS as antigen and adsorbed, specific anti-O1A1 antibodies, whereas O1A1 polysaccharide inhibited this reaction. O1B and O1C LPS differed in all respects tested, including chemical composition, from O1A and O1A1 LPS. Images PMID:2426197

  8. A reexamination of the O1 lipopolysaccharide antigen group of Escherichia coli.

    Science.gov (United States)

    Moll, A; Kusecek, B; Pluschke, G; Morelli, G; Kamke, M; Jann, B; Jann, K; Achtman, M

    1986-08-01

    A total of 64 Escherichia coli strains of the O1 serogroup were tested for the migration pattern of their lipopolysaccharides (LPS) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. O1:K1 and O1:K51 strains of the OMP5 outer membrane protein pattern possessed LPS with a doublet pattern (O1A1) or the lowermost band of the O1A1 doublet (O1A2). O1:K1 strains of the OMP9 pattern possessed LPS referred to as O1A, which corresponded to the uppermost band of the O1A1 doublet pattern. A few O1:K? strains possessed LPS of different migration patterns (O1B and O1C). O1A and O1A1 LPS were indistinguishable by chemical techniques, and both reacted with each of 10 different monoclonal antibodies tested. However, O1A1 had an additional epitope within the additional band in each doublet, as demonstrated by adsorption experiments with hyperimmune rabbit sera followed by Western blotting. Furthermore, purified polysaccharide from O1A bacteria was incapable of inhibition in enzyme-linked immunosorbent assays performed with O1A1 LPS as antigen and adsorbed, specific anti-O1A1 antibodies, whereas O1A1 polysaccharide inhibited this reaction. O1B and O1C LPS differed in all respects tested, including chemical composition, from O1A and O1A1 LPS.

  9. Flexibility of oral cholera vaccine dosing-a randomized controlled trial measuring immune responses following alternative vaccination schedules in a cholera hyper-endemic zone.

    Directory of Open Access Journals (Sweden)

    Suman Kanungo

    2015-03-01

    Full Text Available BACKGROUND: A bivalent killed whole cell oral cholera vaccine has been found to be safe and efficacious for five years in the cholera endemic setting of Kolkata, India, when given in a two dose schedule, two weeks apart. A randomized controlled trial revealed that the immune response was not significantly increased following the second dose compared to that after the first dose. We aimed to evaluate the impact of an extended four week dosing schedule on vibriocidal response. METHODOLOGY/PRINCIPAL FINDINGS: In this double blind randomized controlled non-inferiority trial, 356 Indian, non-pregnant residents aged 1 year or older were randomized to receive two doses of oral cholera vaccine at 14 and 28 day intervals. We compared vibriocidal immune responses between these schedules. Among adults, no significant differences were noted when comparing the rates of seroconversion for V. cholerae O1 Inaba following two dose regimens administered at a 14 day interval (55% vs the 28 day interval (58%. Similarly, no differences in seroconversion were demonstrated in children comparing the 14 (80% and 28 day intervals (77%. Following 14 and 28 day dosing intervals, vibriocidal response rates against V. cholerae O1 Ogawa were 45% and 49% in adults and 73% and 72% in children respectively. Responses were lower for V. cholerae O139, but similar between dosing schedules for adults (20%, 20% and children (28%, 20%. CONCLUSIONS/SIGNIFICANCE: Comparable immune responses and safety profiles between the two dosing schedules support the option for increased flexibility of current OCV dosing. Further operational research using a longer dosing regimen will provide answers to improve implementation and delivery of cholera vaccination in endemic and epidemic outbreak scenarios.

  10. Reduced Susceptibility to Extended-Spectrum β-Lactams in Vibrio cholerae Isolated in Bangladesh

    Science.gov (United States)

    Ceccarelli, Daniela; Alam, Munirul; Huq, Anwar; Colwell, Rita R.

    2016-01-01

    β-lactams are antibiotic molecules able to inhibit cell wall biosynthesis. Among other mechanisms, resistance in Gram-negative bacteria is mostly associated with production of β-lactamase enzymes able to bind and hydrolyze the β-lactam ring. Extended-spectrum β-lactamases extend this ability also to third- and fourth-generation cephalosporins, as well as to carbapenems and monobactams. Vibrio cholerae is the causative agent of epidemic cholera and a public health burden for developing countries like Bangladesh. Although appropriate oral or intravenous rehydration is the therapy of choice for cholera, severe infections and V. cholerae-associated septicemia are treated with antimicrobial drugs, including doxycycline, erythromycin, azithromycin, ciprofloxacin, and/or third-generation cephalosporins. In the years after the introduction of antibiotics in clinical practice, V. cholerae developed resistance to commonly used drugs worldwide mostly through gene acquisition via horizontal gene transfer. Reduced susceptibility of V. cholerae to third-generation cephalosporins has been occasionally documented. However, carbapenemase-producing V. cholerae has been reported at higher rates than resistance to extended-spectrum β-lactams, mainly associated with blaNDM-1 emergence and successful plasmid dissemination. Recent findings suggest limited β-lactam resistance is present in V. cholerae O1 isolates collected during ecological and epidemiological surveillance in Bangladesh. However, a trend to intermediate-susceptibility insurgence was observed. Horizontal gene transfer of β-lactam resistance from enteric pathogens to environmental microorganisms should not be underrated, given the ability of V. cholerae to acquire new genetic information.

  11. Reduced Susceptibility to Extended-Spectrum β-Lactams in Vibrio cholerae Isolated in Bangladesh.

    Science.gov (United States)

    Ceccarelli, Daniela; Alam, Munirul; Huq, Anwar; Colwell, Rita R

    2016-01-01

    β-lactams are antibiotic molecules able to inhibit cell wall biosynthesis. Among other mechanisms, resistance in Gram-negative bacteria is mostly associated with production of β-lactamase enzymes able to bind and hydrolyze the β-lactam ring. Extended-spectrum β-lactamases extend this ability also to third- and fourth-generation cephalosporins, as well as to carbapenems and monobactams. Vibrio cholerae is the causative agent of epidemic cholera and a public health burden for developing countries like Bangladesh. Although appropriate oral or intravenous rehydration is the therapy of choice for cholera, severe infections and V. cholerae-associated septicemia are treated with antimicrobial drugs, including doxycycline, erythromycin, azithromycin, ciprofloxacin, and/or third-generation cephalosporins. In the years after the introduction of antibiotics in clinical practice, V. cholerae developed resistance to commonly used drugs worldwide mostly through gene acquisition via horizontal gene transfer. Reduced susceptibility of V. cholerae to third-generation cephalosporins has been occasionally documented. However, carbapenemase-producing V. cholerae has been reported at higher rates than resistance to extended-spectrum β-lactams, mainly associated with blaNDM-1 emergence and successful plasmid dissemination. Recent findings suggest limited β-lactam resistance is present in V. cholerae O1 isolates collected during ecological and epidemiological surveillance in Bangladesh. However, a trend to intermediate-susceptibility insurgence was observed. Horizontal gene transfer of β-lactam resistance from enteric pathogens to environmental microorganisms should not be underrated, given the ability of V. cholerae to acquire new genetic information.

  12. Reduced susceptibility to extended-spectrum β-lactams in V. cholerae isolated in Bangladesh

    Directory of Open Access Journals (Sweden)

    Daniela Ceccarelli

    2016-10-01

    Full Text Available β-lactams are antibiotic molecules able to inhibit cell wall biosynthesis. Among other mechanisms, resistance in Gram-negative bacteria is mostly associated with production of β-lactamase enzymes able to bind and hydrolyze the β-lactam ring. Extended-spectrum β-lactamases extend this ability also to 3rd- and 4th generation cephalosporins as well as to carbapenems and monobactams. V. cholerae is the causative agent of epidemic cholera and a public health burden for developing countries like Bangladesh. Although appropriate oral or intravenous rehydration is the therapy of choice for cholera, severe infections and V. cholerae associated septicemia are treated with antimicrobial drugs including doxycycline, erythromycin, azithromycin, ciprofloxacin and/or third-generation cephalosporins. In the years after introduction of antibiotics in clinical practice, V. cholerae developed resistance to commonly used drugs worldwide mostly through gene acquisition via horizontal gene transfer. Reduced susceptibility of V. cholerae to third-generation cephalosporins has been occasionally documented. However, carbapenemase-producing V. cholerae has been reported at higher rates than resistance to extended spectrum β-lactams, mainly associated with blaNDM-1 emergence and successful plasmid dissemination. Recent findings suggest limited β-lactam resistance is present in V. cholerae O1 isolates collected during ecological and epidemiological surveillance in Bangladesh. However a trend to intermediate-susceptibility insurgence was observed. Horizontal gene transfer of β-lactam resistance from enteric pathogens to environmental microorganisms should not be underrated, given the ability of V. cholerae to acquire new genetic information.

  13. Comparative genomics of Vibrio cholerae from Haiti, Asia, and Africa.

    Science.gov (United States)

    Reimer, Aleisha R; Van Domselaar, Gary; Stroika, Steven; Walker, Matthew; Kent, Heather; Tarr, Cheryl; Talkington, Deborah; Rowe, Lori; Olsen-Rasmussen, Melissa; Frace, Michael; Sammons, Scott; Dahourou, Georges Anicet; Boncy, Jacques; Smith, Anthony M; Mabon, Philip; Petkau, Aaron; Graham, Morag; Gilmour, Matthew W; Gerner-Smidt, Peter

    2011-11-01

    Cholera was absent from the island of Hispaniola at least a century before an outbreak that began in Haiti in the fall of 2010. Pulsed-field gel electrophoresis (PFGE) analysis of clinical isolates from the Haiti outbreak and recent global travelers returning to the United States showed indistinguishable PFGE fingerprints. To better explore the genetic ancestry of the Haiti outbreak strain, we acquired 23 whole-genome Vibrio cholerae sequences: 9 isolates obtained in Haiti or the Dominican Republic, 12 PFGE pattern-matched isolates linked to Asia or Africa, and 2 nonmatched outliers from the Western Hemisphere. Phylogenies for whole-genome sequences and core genome single-nucleotide polymorphisms showed that the Haiti outbreak strain is genetically related to strains originating in India and Cameroon. However, because no identical genetic match was found among sequenced contemporary isolates, a definitive genetic origin for the outbreak in Haiti remains speculative.

  14. Etiologic characteristics of Vibrio cholerae in Guangdong province in 2009-2013%广东省2009-2013年霍乱弧菌病原学特征分析

    Institute of Scientific and Technical Information of China (English)

    李柏生; 谭海玲; 王多春; 柯碧霞; 陈经雕; 何冬梅; 刘美真; 柯昌文; 张永慧

    2014-01-01

    Objective To analyze the etiologic characteristics of O1/O139 Vibrio cholerae in Guangdong province in 2009-2013.Methods Isolates from cholera cases and from the environment surveillance points were investigated by serological typing,antibiotic susceptibility testings,toxic genes detection and molecular typing to analyze the similarities and differences of the identified species.Results Totally,190 isolations of O1/O139 V.cholerae were obtained from cholera cases (16 strains) and environmental samples (174 strains) in Guangdong province in 2009-2013.The sero-types would include Inaba (3 isolates),Ogawa (7 isolates) and O139 (6 isolates) in all the isolates from the cholera cases.Ten strains from the ctxA positive cases were detected by PCR.Two Ogawa strains carried incomplete CTXΦ phage.Results from the antibiotic susceptibility test indicated that 5 strains were absolutely sensitive to 11 antibiotic discs in vitro,while another 3 strains were simultaneously resistant to 4 antibiotic discs.Except for 2 stains,all the O139 strains from the environment were ctxA negative,detected by PCR.Incomplete CTXΦ phage was found in the Inaba (53 isolates),Ogawa (22 isolates) and O139 (2 isolates),respectively.Results from the antibiotic susceptibility test exhibited that 25 strains were resistant simultaneously to 4 and/or more antibiotic discs,especially the Inaba strains from the seafoods (13 isolates).2 Inaba strains from seafood were simultaneously resistant to 7 antibiotic discs.Results from PFGE molecular typing indicated that the PFGE types digested by Not Ⅰ expressed significant diversity.Inaba and O139 strains from cases were gathered in the same clusters,while the Ogawa strains from cases scattered in different clusters but no significant correlation smong these strains were found.Our results suggested that a distant genetic relationship might exist between these two different sources strains.Conclusion Complex and diverse as the virulence genes and genetic

  15. Cholera toxin production during anaerobic trimethylamine N-oxide respiration is mediated by stringent response in Vibrio cholerae.

    Science.gov (United States)

    Oh, Young Taek; Park, Yongjin; Yoon, Mi Young; Bari, Wasimul; Go, Junhyeok; Min, Kyung Bae; Raskin, David M; Lee, Kang-Mu; Yoon, Sang Sun

    2014-05-09

    As a facultative anaerobe, Vibrio cholerae can grow by anaerobic respiration. Production of cholera toxin (CT), a major virulence factor of V. cholerae, is highly promoted during anaerobic growth using trimethylamine N-oxide (TMAO) as an alternative electron acceptor. Here, we investigated the molecular mechanisms of TMAO-stimulated CT production and uncovered the crucial involvement of stringent response in this process. V. cholerae 7th pandemic strain N16961 produced a significantly elevated level of ppGpp, the bacterial stringent response alarmone, during anaerobic TMAO respiration. Bacterial viability was impaired, and DNA replication was also affected under the same growth condition, further suggesting that stringent response is induced. A ΔrelA ΔspoT ppGpp overproducer strain produced an enhanced level of CT, whereas anaerobic growth via TMAO respiration was severely inhibited. In contrast, a ppGpp-null strain (ΔrelA ΔspoT ΔrelV) grew substantially better, but produced no CT, suggesting that CT production and bacterial growth are inversely regulated in response to ppGpp accumulation. Bacterial capability to produce CT was completely lost when the dksA gene, which encodes a protein that works cooperatively with ppGpp, was deleted. In the ΔdksA mutant, stringent response growth inhibition was alleviated, further supporting the inverse regulation of CT production and anaerobic growth. In vivo virulence of ΔrelA ΔspoT ΔrelV or ΔdksA mutants was significantly attenuated. The ΔrelA ΔspoT mutant maintained virulence when infected with exogenous TMAO despite its defective growth. Together, our results reveal that stringent response is activated under TMAO-stimulated anaerobic growth, and it regulates CT production in a growth-dependent manner in V. cholerae.

  16. Chromosome Segregation in Vibrio cholerae

    OpenAIRE

    Ramachandran, R.; Jha, J.; Chattoraj, DK

    2014-01-01

    The study of chromosome segregation is currently one of the most exciting research frontiers in cell biology. In this review, we discuss our current knowledge of the chromosome segregation process in Vibrio cholerae, based primarily on findings from fluorescence microscopy experiments. This bacterium is of special interest because of its eukaryotic feature of having a divided genome, a feature shared with 10% of known bacteria. We also discuss how the segregation mechanisms of V. cholerae com...

  17. Epidemiological usefulness of changes in hemolytic activity of Vibrio cholerae biotype El Tor during the seventh pandemic.

    OpenAIRE

    Barrett, T J; Blake, P A

    1981-01-01

    Hemolytic Vibrio cholerae biotype El Tor strains were isolated in the United States in 1973 and 1978 after they had supposedly disappeared worldwide during the 1960s and 1970s. We decided to examine the change in prevalence of hemolytic El Tor strains since the beginning of the seventh pandemic and evaluate the usefulness of hemolytic activity as an epidemiological marker. A total of 48 isolates of V. cholerae biotype El Tor isolated in the Eastern Hemisphere between 1960 and 1979, along with...

  18. Improved purification process for cholera toxin and its application to the quantification of residual toxin in cholera vaccines.

    Science.gov (United States)

    Jang, Hyun; Kim, Hyo Seung; Kim, Jeong Ah; Seo, Jin Ho; Carbis, Rodney

    2009-01-01

    A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-microm crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-microm permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH7.0, containing 1.0M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of 3.1 EU/microg of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a G(M1) ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The G(M1) ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

  19. Highly diverse recombining populations of Vibrio cholerae and Vibrio parahaemolyticus in French Mediterranean coastal lagoons

    Directory of Open Access Journals (Sweden)

    Kevin eEsteves

    2015-07-01

    Full Text Available Vibrio parahaemolyticus and Vibrio cholerae are ubiquitous to estuarine and marine environments. These two species can induce infections in humans. Therefore understanding the structure and dynamics of non-pandemic environmental populations in temperate regions, such as Mediterranean coastal systems, is important if we are to evaluate the risks of infection to humans.Environmental isolates of V. cholerae (n=109 and V. parahaemolyticus (n=89 sampled at different dates, stations and water salinities were investigated for virulence genes and by a multilocus sequence-based analysis (MLSA. V. cholerae isolates were all ctxA negative and only one isolate of V. parahaemolyticus displayed trh2 gene. Most Sequence Types (ST corresponded to unique ST isolated at one date or one station. Frequent recombination events were detected among different pathogenic species, V. parahaemolyticus, V. cholerae, Vibrio mimicus and Vibrio metoecus. Recombination had a major impact on the diversification of lineages. The genetic diversity assessed by the number of ST/strain was higher in low salinity conditions for V. parahaemolyticus and V. cholerae whereas the frequency of recombination events in V. cholerae was lower in low salinity. Mediterranean coastal lagoon systems housed V. cholerae and V. parahaemolyticus with genetic diversities equivalent to the worldwide diversity described so far. The presence of STs found in human infections as well as the frequency of recombination events in environmental vibrios populations could predict a potential epidemiological risk.

  20. Highly diverse recombining populations of Vibrio cholerae and Vibrio parahaemolyticus in French Mediterranean coastal lagoons.

    Science.gov (United States)

    Esteves, Kévin; Mosser, Thomas; Aujoulat, Fabien; Hervio-Heath, Dominique; Monfort, Patrick; Jumas-Bilak, Estelle

    2015-01-01

    Vibrio parahaemolyticus and Vibrio cholerae are ubiquitous to estuarine and marine environments. These two species found in Mediterranean coastal systems can induce infections in humans. Environmental isolates of V. cholerae (n = 109) and V. parahaemolyticus (n = 89) sampled at different dates, stations and water salinities were investigated for virulence genes and by a multilocus sequence-based analysis (MLSA). V. cholerae isolates were all ctxA negative and only one isolate of V. parahaemolyticus displayed trh2 gene. Most Sequence Types (ST) corresponded to unique ST isolated at one date or one station. Frequent recombination events were detected among different pathogenic species, V. parahaemolyticus, V. cholerae, Vibrio mimicus, and Vibrio metoecus. Recombination had a major impact on the diversification of lineages. The genetic diversity assessed by the number of ST/strain was higher in low salinity condition for V. parahaemolyticus and V. cholerae whereas the frequency of recombination events in V. cholerae was lower in low salinity condition. Mediterranean coastal lagoon systems housed V. cholerae and V. parahaemolyticus with genetic diversities equivalent to the worldwide diversity described so far. The presence of STs found in human infections as well as the frequency of recombination events in environmental vibrios populations could predict a potential epidemiological risk.

  1. A five-year study on the epidemiological approaches to cholera in Iran

    Science.gov (United States)

    Mafi, Moharam; Goya, Mohammad Mahdi; Hajia, Massoud

    2016-01-01

    Background: Cholera is considered a key indicator of social development but still is reported in various cities of Iran. The present study aimed to analyze the available information regarding cholera outbreaks since 2010 in Iran. Methods: All cases reported to the Center for Disease Control and Prevention of Ministry of Health and Education who had been confirmed as cholera cases by the Health Reference Laboratory, were entered into this study since 2010. A specific spreadsheet was designed to ensure the safe keeping of the patient records. Results: A total of 1522 patients were clinically diagnosed as cholera with laboratory confirmation over the study period. Cholera was detected in 26 Provinces and 115 cities during this period. Mean age of the patients was 35.1±17, both the Inaba and Ogawa strains were isolated. The highest mortality and the morbidity rate was 1.98% in 2013. The most cholera prevalent provinces in order of frequency were Baluchistan, Alborz, Gilan, Golestan and Qom, as well as Tehran. Inaba serotype was the most common cause of mortality and morbidity in 2013. Conclusion: These findings indicate significant outbreaks of cholera in some of the provinces of Iran and warrant appropriate treatment and preventive measures. PMID:27757199

  2. EROTYPE IDENTIFICATION OF VIBRIO CHOLERAE BACTERIAWHICH ISOLATED FROM ICE AMONGTUBE AND CUBE ICE TYPE IN FOOD AND BEVERAGES SELLER AT DENPASAR CITY, BALI

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    IGP Dhinarananta

    2014-01-01

    Full Text Available Cholera is a type of watery diarrhea with specific sign stool containing mucus which resembles rice water. Cholera caused by gram negative bacteria Vibrio cholerae (V.Cholerae. The transmissions of bacteria were through a contaminated food or water.Bali is an international tourism destination with tropical weather where ice is widelyused in food and beverage which bring a risk of cholera through a contaminated ice.Iceshave a risk of bacterial contamination whether from the making and the usage process.Type of ice that widely used were cube and tube ice which each of them have a differentin making and usage process. The purpose of this study is to obtain the contamination ofV.cholera in cube and tube ice. The method of this study is descriptive observationalstudy with quota sampling technique. Sample were obtained from a restaurants andstreet vendor which use a block and tube ice with total 10 sample and 5 for each type ofice.Sample then cultured in Alkaline Peptone Water(APW and Thiosulfate Citrate Bilesalt Sucrose(TCBS agar. Bacteriacolony then identified using a gram staining andLatex Serotyping. The result are 3 over 5 (60% sample of cube ice contaminated byV.cholera O1 Inaba serotype and 3 over 5 (60% sample of tube ice contaminated byV.cholera O1 Inaba serotype.

  3. Genetic Profile of IS1004 among Environmental Vibrio cholerae Isolated from Surface Water Sources in Iran

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    Bita Bakhshi B

    2013-03-01

    Full Text Available AbstractBackground and objective: Vibrio cholerae includes toxigenic and non-toxigenic serotypes. Non O1-non O139 serotypes are non toxigenic and do not have any role in cholera epidemics or pandemics all over the world. Different typing methods are widely used for molecular epidemiological investigations of clinical and environmental V. cholerae. The aim of this study is to investigate the genetic relatedness of environmental isolates of this bacterium using IS1004 profiling as an epidemiological marker. Materials and methods: Environmental samples collected from surface water sources in Tehran and cultured of TCBS agar after filtration. One single colony on TCBS was selected and cultured on BHI agar after which the cultures were used for biochemical diagnostic tests and serogroupings. DNA was isolated and used for PCR confirmation of V. cholerae isolates and wbeT gene. Genetic relatedness of isolates was determined using southern blot analysis. Results: From total 20 environmental V. cholerae identified in this study no wbeT gene was detected for the isolates. A total of 7 different banding patterns were obtained for the isolates while other 13 isolates identified as non-typeable by this method. Comparison with our previous studies indicated no identical pattern with clinical V. cholerae isolates. Conclusion: Differences in the banding pattern of IS1004 revealed a high heterogeneity among the isolates from surface water sources in Iran while these heterogenic isolates do not have any genetic relatedness with clinical isolates.

  4. Isolation and Antimicrobial Sensitivity Test of A Vibrio cholerae Strain from Penaeus vannamei with White Fecal Syndrome%南美白对虾白便综合征病原霍乱弧菌的分离与药敏试验

    Institute of Scientific and Technical Information of China (English)

    曹海鹏; 温乐夫; 周桂娴; 张帆; 何珊

    2016-01-01

    In order to confirm the bacterial pathogen of Penaeus vannamei white fecal syndrome and its an-timicrobial sensitivity,a pathogenic strain BB31 was isolated from white fecal syndrome Penaeus van-namei using traditional methods,identified by API 20E Gram negative bacterial identification system,1 6 S rDNA sequence and phylogenetic analysis.Its antimicrobial susceptibility was also analyzed by Kirby-Bauer test.The results showed that strain BB31 was a Vibrio cholerae strain with its GenBank accession No.KF446244,its 1 6 S rDNA sequence had a 99%-100% homology with those of Vibrio strains,and showed a closest relationship to that of V .cholerae strain RD1 with GenBank accession No.KF307775. Strain BB31 exhibited a high sensitivity to tetracycline,gentamycin,enrofloxacin,ofloxacin and norfloxa-cin,showed a resistance against sulfametoxydiazine, compound sulfamethoxazole,furazolidone.This study confirmed Vibrio cholerae as a pathogen of white fecal syndrome Penaeus vannamei ,and common fishery antimicrobial such as gentamycin and norfloxacin may be regarded as useful antimicrobial in the control of Penaeus vannamei white fecal syndrome caused by Vibrio cholerae .%为确定南美白对虾白便综合征的病原菌及其药物敏感性,采用传统方法从患白便综合征的南美白对虾肝胰腺中分离到病原菌菌株 BB31,结合 API 20E 革兰阴性菌鉴定系统、16 S rDNA 序列及聚类分析对菌株 BB31进行了鉴定,并通过纸片琼脂扩散法对菌株 BB31的药物敏感性进行了测定。结果表明,菌株BB31为霍乱弧菌(GenBank 登录号:KF446244),其16 S rDNA 序列与 GenBank 数据库中弧菌菌株的16 S rDNA 序列有99%~100%的同源性,而且与霍乱弧菌菌株 RD1(GenBank 登录号:KF307775)的亲缘性最近。菌株 BB31对四环素、庆大霉素、恩诺沙星、氧氟沙星、诺氟沙星等抗生素的敏感性高,对磺胺甲氧嘧啶、呋喃唑酮、复方新诺明等抗生素不敏感。本研究证实霍乱

  5. The burden of diarrhoea, shigellosis, and cholera in North Jakarta, Indonesia: findings from 24 months surveillance

    Directory of Open Access Journals (Sweden)

    Lee Hyejon

    2005-10-01

    Full Text Available Abstract Background In preparation of vaccines trials to estimate protection against shigellosis and cholera we conducted a two-year community-based surveillance study in an impoverished area of North Jakarta which provided updated information on the disease burden in the area. Methods We conducted a two-year community-based surveillance study from August 2001 to July 2003 in an impoverished area of North Jakarta to assess the burden of diarrhoea, shigellosis, and cholera. At participating health care providers, a case report form was completed and stool sample collected from cases presenting with diarrhoea. Results Infants had the highest incidences of diarrhoea (759/1 000/year and cholera (4/1 000/year. Diarrhea incidence was significantly higher in boys under 5 years (387/1 000/year than girls under 5 years (309/1 000/year; p Shigella flexneri was the most common Shigella species isolated and 73% to 95% of these isolates were resistant to ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol and tetracycline but remain susceptible to nalidixic acid, ciprofloxacin, and ceftriaxone. We found an overall incidence of cholera of 0.5/1 000/year. Cholera was most common in children, with the highest incidence at 4/1 000/year in those less than 1 year of age. Of the 154 V. cholerae O1 isolates, 89 (58% were of the El Tor Ogawa serotype and 65 (42% were El Tor Inaba. Thirty-four percent of patients with cholera were intravenously rehydrated and 22% required hospitalization. V. parahaemolyticus infections were detected sporadically but increased from July 2002 onwards. Conclusion Diarrhoea causes a heavy public health burden in Jakarta particularly in young children. The impact of shigellosis is exacerbated by the threat of antimicrobial resistance, whereas that of cholera is aggravated by its severe manifestations.

  6. Role of Indole Production on Virulence of Vibrio cholerae Using Galleria mellonella Larvae Model.

    Science.gov (United States)

    Nuidate, Taiyeebah; Tansila, Natta; Saengkerdsub, Suwat; Kongreung, Jetnaphang; Bakkiyaraj, Dhamodharan; Vuddhakul, Varaporn

    2016-09-01

    Cell to cell communication facilitated by chemical signals plays crucial roles in regulating various cellular functions in bacteria. Indole, one such signaling molecule has been demonstrated to control various bacterial phenotypes such as biofilm formation and virulence in diverse bacteria including Vibrio cholerae. The present study explores some key factors involved in indole production and the subsequent pathogenesis of V. cholerae. Indole production was higher at 37 °C than at 30 °C, although the growth at 37 °C was slightly higher. A positive correlation was observed between indole production and biofilm formation in V. cholerae. Maximum indole production was detected at pH 7. There was no significant difference in indole production between clinical and environmental V. cholerae isolates, although indole production in one environmental isolate was significantly different. Both growth and indole production showed relevant changes with differences in salinity. An indole negative mutant strain was constructed using transposon mutagenesis and the direct effect of indole on the virulence of V. cholerae was evaluated using Galleria mellonella larvae model. Comparison to the wild type strain, the mutant significantly reduced the mortality of G. mellonella larvae which regained its virulence after complementation with exogenous indole. A gene involved in indole production and the virulence of V. cholerae was identified.

  7. Binding of cholera toxin to Giardia lamblia.

    OpenAIRE

    McCardell, B. A.; Madden, J M; Stanfield, J T; Tall, B D; Stephens, M. J.

    1987-01-01

    Binding of cholera toxin to Giardia lamblia was demonstrated by two slightly different methods: an immunofluorescence technique using antibody to cholera toxin and anti-rabbit immunoglobulin G conjugated to fluorescein isothiocyanate, and a one-step fluorescence method in which G. lamblia was incubated with the B subunit of cholera toxin conjugated to fluorescein isothiocyanate.

  8. Agent-based modelling of cholera diffusion

    NARCIS (Netherlands)

    Augustijn, Ellen-Wien; Doldersum, Tom; Useya, Juliana; Augustijn, Denie

    2016-01-01

    This paper introduces a spatially explicit agent-based simulation model for micro-scale cholera diffusion. The model simulates both an environmental reservoir of naturally occurring V. cholerae bacteria and hyperinfectious V. cholerae. Objective of the research is to test if runoff from open refuse

  9. Agent-based modelling of cholera diffusion

    NARCIS (Netherlands)

    Augustijn, Ellen-Wien; Doldersum, Tom; Useya, Juliana; Augustijn, Denie

    2016-01-01

    This paper introduces a spatially explicit agent-based simulation model for micro-scale cholera diffusion. The model simulates both an environmental reservoir of naturally occurring V.cholerae bacteria and hyperinfectious V. cholerae. Objective of the research is to test if runoff from open refuse d

  10. Chlorination of Household Drinking Water Among Cholera Patients' Households to Prevent Transmission of Toxigenic Vibrio cholerae in Dhaka, Bangladesh: CHoBI7 Trial.

    Science.gov (United States)

    Rashid, Mahamud-Ur; George, Christine Marie; Monira, Shirajum; Mahmud, Toslim; Rahman, Zillur; Mustafiz, Munshi; Saif-Ur-Rahman, K M; Parvin, Tahmina; Bhuyian, Sazzadul Islam; Zohura, Fatema; Begum, Farzana; Biswas, Shwapon Kumar; Akhter, Shamima; Zhang, Xiaotong; Sack, David; Sack, R Bradley; Alam, Munirul

    2016-12-07

    Household members of cholera patients are at a 100 times higher risk of cholera infections than the general population because of shared contaminated drinking water sources and secondary transmission through poor household hygiene practices. In this study, we investigated the bactericidal concentration of free chlorine required to inactivate Vibrio cholerae in household drinking water in Dhaka, Bangladesh. In laboratory experiments, we found that the concentrations of free chlorine required to inactivate 10(5) colony-forming units (CFU)/mL of V. cholerae serogroups O1 and O139 were 0.1 mg/L and 0.2 mg/L, respectively. The concentration of free chlorine generated by a single chlorine tablet (sodium dichloroisocyanurate [33 mg]) after a 30-minute reaction time in a 10-L sealed vessel containing Dhaka city municipal supply water was 1.8 mg/L; and the concentration declined to 0.26 mg/L after 24 hours. In field measurements, water collected from 165 households enrolled in a randomized controlled trial (RCT) of a chlorine and handwashing with soap intervention (Cholera-Hospital-Based-Intervention-for-7-Days [CHoBI7]), we observed significantly higher free chlorine concentrations in the 82 intervention arm households (mean = 1.12 mg/L, standard deviation [SD] = 0.52, range = 0.07-2.6 mg/L) compared with the 83 control households (0.017 mg/L, SD = 0.01, range = 0-0.06 mg/L) (P < 0.001) during spot check visits. These findings suggest that point-of-use chlorine tablets present an effective approach to inactivate V. cholerae from drinking water in households of cholera patients in Dhaka city. This result is consistent with the findings from the RCT of CHoBI7 which found that this intervention led to a significant reduction in symptomatic cholera infections among household members of cholera patients and no stored drinking water samples with detectable V. cholerae.

  11. Genome sequencing reveals unique mutations in characteristic metabolic pathways and the transfer of virulence genes between V. mimicus and V. cholerae.

    Science.gov (United States)

    Wang, Duochun; Wang, Haiyin; Zhou, Yanyan; Zhang, Qiuxiang; Zhang, Fanfei; Du, Pengcheng; Wang, Shujing; Chen, Chen; Kan, Biao

    2011-01-01

    Vibrio mimicus, the species most similar to V. cholerae, is a microbe present in the natural environmental and sometimes causes diarrhea and internal infections in humans. It shows similar phenotypes to V. cholerae but differs in some biochemical characteristics. The molecular mechanisms underlying the differences in biochemical metabolism between V. mimicus and V. cholerae are currently unclear. Several V. mimicus isolates have been found that carry cholera toxin genes (ctxAB) and cause cholera-like diarrhea in humans. Here, the genome of the V. mimicus isolate SX-4, which carries an intact CTX element, was sequenced and annotated. Analysis of its genome, together with those of other Vibrio species, revealed extensive differences within the Vibrionaceae. Common mutations in gene clusters involved in three biochemical metabolism pathways that are used for discrimination between V. mimicus and V. cholerae were found in V. mimicus strains. We also constructed detailed genomic structures and evolution maps for the general types of genomic drift associated with pathogenic characters in polysaccharides, CTX elements and toxin co-regulated pilus (TCP) gene clusters. Overall, the whole-genome sequencing of the V. mimicus strain carrying the cholera toxin gene provides detailed information for understanding genomic differences among Vibrio spp. V. mimicus has a large number of diverse gene and nucleotide differences from its nearest neighbor, V. cholerae. The observed mutations in the characteristic metabolism pathways may indicate different adaptations to different niches for these species and may be caused by ancient events in evolution before the divergence of V. cholerae and V. mimicus. Horizontal transfers of virulence-related genes from an uncommon clone of V. cholerae, rather than the seventh pandemic strains, have generated the pathogenic V. mimicus strain carrying cholera toxin genes.

  12. Architecture of the superintegron in Vibrio cholerae: identification of core and unique genes [v1; ref status: indexed, http://f1000r.es/w6

    Directory of Open Access Journals (Sweden)

    Michel A Marin

    2013-02-01

    Full Text Available Background: Vibrio cholerae, the etiologic agent of cholera, is indigenous to aquatic environments. The V. cholerae genome consists of two chromosomes; the smallest of these harbors a large gene capture and excision system called the superintegron (SI, of ~120 kbp. The flexible nature of the SI that results from gene cassette capture, deletion and rearrangement is thought to make it a hotspot of V. cholerae diversity, but beyond the basic structure it is not clear if there is a core genome in the SI and if so how it is structured. The aim of this study was to explore the core genome structure and the differences in gene content among strains of V. cholerae. Methods: From the complete genomes of seven V. cholerae and one Vibrio mimicus representative strains, we recovered the SI sequences based on the locations of the structural gene IntI4 and the V. cholerae repeats. Analysis of the pangenome, including cluster analysis of functional genes, pangenome profile analysis, genetic variation analysis of functional genes, strain evolution analysis and function enrichment analysis of gene clusters, was performed using a pangenome analysis pipeline in addition to the R scripts, splitsTree4 and genoPlotR. Results and conclusions: Here, we reveal the genetic architecture of the V. cholerae SI. It contains eight core genes when V. mimicus is included and 21 core genes when only V. cholerae strains are considered; many of them are present in several copies. The V. cholerae SI has an open pangenome, which means that V. cholerae may be able to import new gene cassettes to SI. The set of dispensable SI genes is influenced by the niche and type species. The core genes are distributed along the SI, apparently without a position effect.

  13. Lack of Outer Membrane Protein A Enhances the Release of Outer Membrane Vesicles and Survival of Vibrio cholerae and Suppresses Viability of Acanthamoeba castellanii

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    Soni Priya Valeru

    2014-01-01

    Full Text Available Vibrio cholerae, the causative agent of the diarrhoeal disease cholera, survives in aquatic environments. The bacterium has developed a survival strategy to grow and survive inside Acanthamoeba castellanii. It has been shown that V. cholerae expresses outer membrane proteins as virulence factors playing a role in the adherence to interacted host cells. This study examined the role of outer membrane protein A (OmpA and outer membrane vesicles (OMVs in survival of V. cholerae alone and during its interaction with A. castellanii. The results showed that an OmpA mutant of V. cholerae survived longer than wild-type V. cholerae when cultivated alone. Cocultivation with A. castellanii enhanced the survival of both bacterial strains and OmpA protein exhibited no effect on attachment, engulfment, and survival inside the amoebae. However, cocultivation of the OmpA mutant of V. cholerae decreased the viability of A. castellanii and this bacterial strain released more OMVs than wild-type V. cholerae. Surprisingly, treatment of amoeba cells with OMVs isolated from the OmpA mutant significantly decreased viable counts of the amoeba cells. In conclusion, the results might highlight a regulating rule for OmpA in survival of V. cholerae and OMVs as a potent virulence factor for this bacterium towards eukaryotes in the environment.

  14. Design of a multiplex PCR method for detection of toxigenic- pathogenic inVibrio cholerae

    Institute of Scientific and Technical Information of China (English)

    Imani Fooladi AA; Iman Islamieh D; Hosseini Doust R; Karami A; Marashi SM

    2013-01-01

    Objective:To study virulence and regulatory genes (hlyA,ctxB,tcpI) in clinical strains ofVibrio cholerae (V. cholerae), simultaneously.Methods:Three important genes,tcpI,hlyA andctxB were used for detection of toxigenic and pathogenicV. cholera by chain reaction assay method. Results:According to the results of thePCR, the incidence ofhlyA,tcpI, andctxB genes in clinical isolates was obtained as94.7% (72 sample),90.8% (69 sample), and92.1% (70 sample), respectively.Five strains possessed all genes exceptctxB, six strains possessed all genes except tcpI, four strains possessed all genes excepthlyA, one strain possessed onlyhlyA and60 strains contained a combination of three genes,IncludinghlyA,ctxB andtcpI.Conclusions:Result show that this method could be reliable to detect toxigenic-pathogenic strains ofV. cholerae in Iran.

  15. Biological Activity of Recombinant Accessory Cholerae Enterotoxin (Ace on Rabbit Ileal Loops and Antibacterial Assay

    Directory of Open Access Journals (Sweden)

    Shaghayegh Anvari

    2012-01-01

    Full Text Available Objective: Vibrio cholerae (V. cholerae causes a potentially lethal disease named cholera. The cholera enterotoxin (CT is a major virulence factor of V. cholerae. In addition to CT, V. cholerae produces other putative toxins, such as the zonula occludens toxin (Zot and accessory cholera enterotoxin (Ace. The ace gene is the third gene of the V. cholerae virulence cassette. The Ace toxin alters ion transport, causes fluid accumulation in ligated rabbit ileal loops, and is a cause of mild diarrhea. The aim of this study is the cloning and overexpression of the ace gene into Escherichia coli (E. coli and determination of some characteristics of the recombinant Ace protein.Materials and Methods: In this experimental study, the ace gene was amplified from V. cholerae strain 62013, then cloned in a pET28a expression vector and transformed into an E. coli (DH5 α host strain. Subsequently, the recombinant vector was retransformed into E. coli BL21 for expression, induced by isopropythio-β-D-galctoside (IPTG at a different concentration, and examined by SDS-PAGE and Western blot. A rabbit ileal loop experiment was conducted. Antibacterial activity of the Ace protein was assessed for E. coli, Stapylococcus aureus (S. aureus, and Pseudomonas aeruginosa (P. aeruginosa.Results: The recombinant Ace protein with a molecular weight of 18 kDa (dimeric form was expressed in E. coli BL21. The Ace protein showed poor staining with Coomassie blue stain, but stained efficiently with silver stain. Western blot analysis showed that the recombinant Ace protein reacted with rabbit anti-V. cholerae polyclonal antibody. The Ace protein had antibacterial activity at a concentration of ≥200 μg/ml and caused significant fluid accumulation in the ligated rabbit ileal loop test.Conclusion: This study described an E. coli cloning and expression system (E. coli BL21- pET-28a-ace for the Ace protein of V. cholerae. We confirmed the antibacterial properties and enterotoxin

  16. Enhanced Detection of Vibrio Cholerae in Oyster Homogenate Based on Centrifugal Removal of Inhibitory Agents

    Science.gov (United States)

    Alexander, Donita; DePaola, Angelo; Young, Ronald B.

    1998-01-01

    The disease cholera, caused by Vibrio cholerae, has been associated with consumption of contaminated seafood, including raw oysters. Detection of V. cholerae in foods typically involves blending the oysters, diluting the homogenate in alkaline peptone water (APW), overnight enrichment, and isolation on selective agar. Unfortunately, the oyster homogenate must be diluted to large volumes because lower dilutions inhibit the growth of V. cholerae. The goals of this study were to develop an alternative to large dilutions and to evaluate the basis for the inhibition observed in lower dilutions of oyster homogenates. Centrifugation of oyster homogenates at 10,000 x g for 15 min, followed by enrichment of the resulting pellet in APW, was found to eliminate the inhibition of V. cholerae growth. Inhibition appears not to be due to competing microflora but to a component(s) released when V. cholerae grows in the presence of oyster homogenate. The inhibitory component(s) kills the V. cholerae after the cell concentration reaches > 10(exp 8) cells/mL, rather than initially preventing their growth. The pH also declines from 8.0 to 5.5 during this period; however, the pH decline by itself appears not to cause V. cholerae death. Seven strains of V. cholerae (01 and non-01) and two strains of V. vulnificus were susceptible to the inhibitory agent(s). However, other Vibrio and non-Vibrio species tested were not inhibited by the oyster homogenates. Based on digestion of oyster homogenates with pronase, trypsin and lipase, the inhibitory reaction involves a protein(s). In a preliminary trial with oyster homogenate seeded with 1 cfu/g of V. cholerae, the modified centrifugation technique detected a slightly higher percentage of samples at a 1:10 dilution than the standard FDA Bacteriological Analytical Method (BAM) detected in uncentrifuged oyster homogenate at a 1:100 dilution. V. cholerae in seeded samples could also be detected more frequently by the modified centrifugation method

  17. The Dynamics of Genetic Interactions between Vibrio metoecus and Vibrio cholerae, Two Close Relatives Co-Occurring in the Environment.

    Science.gov (United States)

    Orata, Fabini D; Kirchberger, Paul C; Méheust, Raphaël; Barlow, E Jed; Tarr, Cheryl L; Boucher, Yan

    2015-10-09

    Vibrio metoecus is the closest relative of Vibrio cholerae, the causative agent of the potent diarrheal disease cholera. Although the pathogenic potential of this new species is yet to be studied in depth, it has been co-isolated with V. cholerae in coastal waters and found in clinical specimens in the United States. We used these two organisms to investigate the genetic interaction between closely related species in their natural environment. The genomes of 20 V. cholerae and 4 V. metoecus strains isolated from a brackish coastal pond on the US east coast, as well as 4 clinical V. metoecus strains were sequenced and compared with reference strains. Whole genome comparison shows 86-87% average nucleotide identity (ANI) in their core genes between the two species. On the other hand, the chromosomal integron, which occupies approximately 3% of their genomes, shows higher conservation in ANI between species than any other region of their genomes. The ANI of 93-94% observed in this region is not significantly greater within than between species, meaning that it does not follow species boundaries. Vibrio metoecus does not encode toxigenic V. cholerae major virulence factors, the cholera toxin and toxin-coregulated pilus. However, some of the pathogenicity islands found in pandemic V. cholerae were either present in the common ancestor it shares with V. metoecus, or acquired by clinical and environmental V. metoecus in partial fragments. The virulence factors of V. cholerae are therefore both more ancient and more widespread than previously believed. There is high interspecies recombination in the core genome, which has been detected in 24% of the single-copy core genes, including genes involved in pathogenicity. Vibrio metoecus was six times more often the recipient of DNA from V. cholerae as it was the donor, indicating a strong bias in the direction of gene transfer in the environment.

  18. Chromosome segregation in Vibrio cholerae.

    Science.gov (United States)

    Ramachandran, Revathy; Jha, Jyoti; Chattoraj, Dhruba K

    2014-01-01

    The study of chromosome segregation is currently one of the most exciting research frontiers in cell biology. In this review, we discuss our current knowledge of the chromosome segregation process in Vibrio cholerae, based primarily on findings from fluorescence microscopy experiments. This bacterium is of special interest because of its eukaryotic feature of having a divided genome, a feature shared with 10% of known bacteria. We also discuss how the segregation mechanisms of V. cholerae compare with those in other bacteria, and highlight some of the remaining questions regarding the process of bacterial chromosome segregation.

  19. Epidemiology of cholera in the Philippines.

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    Anna Lena Lopez

    2015-01-01

    Full Text Available Despite being a cholera-endemic country, data on cholera in the Philippines remain sparse. Knowing the areas where cholera is known to occur and the factors that lead to its occurrence will assist in planning preventive measures and disaster mitigation.Using sentinel surveillance data, PubMed and ProMED searches covering information from 2008-2013 and event-based surveillance reports from 2010-2013, we assessed the epidemiology of cholera in the Philippines. Using spatial log regression, we assessed the role of water, sanitation and population density on the incidence of cholera.We identified 12 articles from ProMED and none from PubMed that reported on cholera in the Philippines from 2008 to 2013. Data from ProMed and surveillance revealed 42,071 suspected and confirmed cholera cases reported from 2008 to 2013, among which only 5,006 were confirmed. 38 (47% of 81 provinces and metropolitan regions reported at least one confirmed case of cholera and 32 (40% reported at least one suspected case. The overall case fatality ratio in sentinel sites was 0.62%, but was 2% in outbreaks. All age groups were affected. Using both confirmed and suspected cholera cases, the average annual incidence in 2010-2013 was 9.1 per 100,000 population. Poor access to improved sanitation was consistently associated with higher cholera incidence. Paradoxically, access to improved water sources was associated with higher cholera incidence using both suspected and confirmed cholera data sources. This finding may have been due to the breakdown in the infrastructure and non-chlorination of water supplies, emphasizing the need to maintain public water systems.Our findings confirm that cholera affects a large proportion of the provinces in the country. Identifying areas most at risk for cholera will support the development and implementation of policies to minimize the morbidity and mortality due to this disease.

  20. Cholera toxin-B (ctxB) antigen expressing Salmonella Typhimurium polyvalent vaccine exerts protective immune response against Vibrio cholerae infection.

    Science.gov (United States)

    Vishwakarma, Vikalp; Sahoo, Sushree Sangita; Das, Susmita; Ray, Shilpa; Hardt, Wolf-Dietrich; Suar, Mrutyunjay

    2015-04-08

    Live attenuated vaccines are cost effective approach for preventing a broad range of infectious diseases, and thus are of great interest. However, immune-defects can predispose the patient to infections by the vaccine candidate itself. So far, few live vaccine candidates have been designed specifically for immune compromised individuals. Recently, we reported a new Salmonella Typhimurium Z234-vaccine strain (Periaswamy et al., PLoS ONE 2012;7:e45433), which was specifically attenuated in the NADPH-oxidase deficient host. In the present study, the Z234-vaccine strain was further engineered to express heterologous antigen (Vibrio cholerae toxin antigen subunit-B, i.e. CtxB) with the intention of creating a vector for simultaneous protection against Cholera and Salmonellosis. The primary aim of this study was to ensure the expression of CtxB antigen by the recombinant vaccine strain Z234-pMS101. The antigen CtxB was expressed through Z234 as a fusion protein with N-terminal signal sequence of Salmonella outer protein (SopE), an effector protein from Salmonella under the control of SopE promoter. The CtxB-expressing plasmid construct pMS101 (pM968-pSopE-ctxB) was found to be stable both in vitro and in vivo. In an oral mouse infection model, the vaccine strain Z234-pMS101 efficiently colonized the host gut. The extent of protection was confirmed after challenging the immunized hosts with live V. cholerae. Vaccinated mice showed reduced gut colonization by V. cholerae. Further assessment of immunological parameters supported the possibility of conferring effective immune response by Z234-pMS101 vaccine strain. Overall, the Z234-pMS101 vaccine strain showed potential as a promising polyvalent vaccine candidate to protect against S. Typhimurium and V. cholerae infection simultaneously.

  1. RpoS controls the Vibrio cholerae mucosal escape response.

    Directory of Open Access Journals (Sweden)

    Alex Toftgaard Nielsen

    2006-10-01

    Full Text Available Vibrio cholerae causes a severe diarrhoeal disease by secreting a toxin during colonization of the epithelium in the small intestine. Whereas the initial steps of the infectious process have been intensively studied, the last phases have received little attention. Confocal microscopy of V. cholerae O1-infected rabbit ileal loops captured a distinctive stage in the infectious process: 12 h post-inoculation, bacteria detach from the epithelial surface and move into the fluid-filled lumen. Designated the "mucosal escape response," this phenomenon requires RpoS, the stationary phase alternative sigma factor. Quantitative in vivo localization assays corroborated the rpoS phenotype and showed that it also requires HapR. Expression profiling of bacteria isolated from ileal loop fluid and mucus demonstrated a significant RpoS-dependent upregulation of many chemotaxis and motility genes coincident with the emigration of bacteria from the epithelial surface. In stationary phase cultures, RpoS was also required for upregulation of chemotaxis and motility genes, for production of flagella, and for movement of bacteria across low nutrient swarm plates. The hapR mutant produced near-normal numbers of flagellated cells, but was significantly less motile than the wild-type parent. During in vitro growth under virulence-inducing conditions, the rpoS mutant produced 10- to 100-fold more cholera toxin than the wild-type parent. Although the rpoS mutant caused only a small over-expression of the genes encoding cholera toxin in the ileal loop, it resulted in a 30% increase in fluid accumulation compared to the wild-type. Together, these results show that the mucosal escape response is orchestrated by an RpoS-dependent genetic program that activates chemotaxis and motility functions. This may furthermore coincide with reduced virulence gene expression, thus preparing the organism for the next stage in its life cycle.

  2. Genomic analysis of immune response against Vibrio cholerae hemolysin in Caenorhabditis elegans.

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    Surasri N Sahu

    Full Text Available Vibrio cholerae cytolysin (VCC is among the accessory V. cholerae virulence factors that may contribute to disease pathogenesis in humans. VCC, encoded by hlyA gene, belongs to the most common class of bacterial toxins, known as pore-forming toxins (PFTs. V. cholerae infects and kills Caenorhabditis elegans via cholerae toxin independent manner. VCC is required for the lethality, growth retardation and intestinal cell vacuolation during the infection. However, little is known about the host gene expression responses against VCC. To address this question we performed a microarray study in C. elegans exposed to V. cholerae strains with intact and deleted hlyA genes.Many of the VCC regulated genes identified, including C-type lectins, Prion-like (glutamine [Q]/asparagine [N]-rich-domain containing genes, genes regulated by insulin/IGF-1-mediated signaling (IIS pathway, were previously report