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Sample records for cholerae o1 strains

  1. Purification and characterization of pili of a Vibrio cholerae non-O1 strain.

    OpenAIRE

    Yamashiro, T.; Nakasone, N; Iwanaga, M

    1993-01-01

    Pili of the Vibrio cholerae non-O1 strain V10 were purified and characterized. The V10 pili were physicochemically and immunologically different from those of the previously reported V. cholerae non-O1 strain S7, although the pili of the two strains had homologous N-terminal amino acid sequences. V10 plus antigen was detected only in V. cholerae non-O1 strains.

  2. 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

  3. Pulmonary Cholera Due to Infection with a Non-O1 Vibrio cholerae Strain

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    Shannon, Jack D.; Kimbrough, Robert C.

    2006-01-01

    We present 2 cases of primary pulmonary non-O1 Vibrio cholerae infection. We believe that these are the first documented cases of primary pulmonary infection due to this organism from a freshwater source.

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

    Science.gov (United States)

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

    Science.gov (United States)

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

    2016-01-01

    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. PMID:26956038

  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. Genome Sequence and Comparative Genomics Analysis of a Vibrio cholerae O1 Strain Isolated from a Cholera Patient in Malaysia

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    Osama, Abdulrazak; Gan, Han Ming; Teh, Cindy Shuan Ju; Yap, Kien-Pong

    2012-01-01

    The genome sequence analysis of a clinical Vibrio cholerae VC35 strain from an outbreak case in Malaysia indicates multiple genes involved in host adaptation and a novel Na+-driven multidrug efflux pump-coding gene in the genome of Vibrio cholerae with the highest similarity to VMA_001754 of Vibrio mimicus VMA223. PMID:23209200

  9. Genome Sequence and Comparative Genomics Analysis of a Vibrio cholerae O1 Strain Isolated from a Cholera Patient in Malaysia

    OpenAIRE

    Osama, Abdulrazak; Gan, Han Ming; Teh, Cindy Shuan Ju; Yap, Kien-Pong; Thong, Kwai-Lin

    2012-01-01

    The genome sequence analysis of a clinical Vibrio cholerae VC35 strain from an outbreak case in Malaysia indicates multiple genes involved in host adaptation and a novel Na+-driven multidrug efflux pump-coding gene in the genome of Vibrio cholerae with the highest similarity to VMA_001754 of Vibrio mimicus VMA223.

  10. 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

  11. Vibrio cholerae O1 lineages driving cholera outbreaks during seventh cholera pandemic in Ghana.

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    Thompson, Cristiane C; Freitas, Fernanda S; Marin, Michel A; Fonseca, Erica L; Okeke, Iruka N; Vicente, Ana Carolina P

    2011-12-01

    In recent years, the frequency of cholera epidemics across Africa has increased significantly with thousands of people dying each year. However, there still exists a lack of information concerning the Vibrio cholerae O1 lineages driving early and contemporary epidemics since the seventh cholera pandemic started in the continent. This compromises the understanding of the forces determining the epidemiology of cholera in Africa and its control. This study aimed to analyze a collection of V. cholerae O1 strains from the beginning of the seventh cholera pandemic in Ghana and to compare them with recent isolates to understand the evolution of the cholera epidemic in Ghana. V. cholerae O1 strains were characterized by means of Multilocus Sequence Analysis (MLSA), genes from the virulence core genome (VCG), and genes related to the choleragenic phenotype. Our results revealed two major clusters of Ghanaian V. cholerae O1 strains, El Tor and Amazonia/Ghana. Concerning the virulence genes, all strains harbored the set of VCG and most were positive for VSP-II genomic island. The ctxB gene of the contemporary strains was characterized as Altered El Tor. The strains from 1970 to 1980 were susceptible to all antibiotics tested, except for the Amazonia/Ghana cluster that was resistant to aminoglycosides and carried the class 2 integron with the sat2-aadA1 arrangement. This study showed that distinct V. cholerae O1 were the determinants of cholera outbreaks in Ghana. Thus, in endemic regions, such as Africa, cholera can be caused by various V. cholerae O1 genotypes. PMID:21896336

  12. Genetic traits of Vibrio cholerae O1 Haitian isolates that are absent in contemporary strains from Kolkata, India.

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    Priyanka Ghosh

    Full Text Available The world's worst cholera epidemic in Haiti (2010 coerced to trace the origin and dissemination of the causative agent Vibrio cholerae O1 for proper management of cholera. Sequence analysis of the Haitian strain showed several variations in the genes encoding cholera toxin B subunit (ctxB; toxin-co-regulated pilus (tcpA, repeat in toxins (rtxA, quinolone resistance-determining region (QRDR of gyrase A (gyrA, rstB of RS element along with the change in the number of repeat sequences at the promoter region of ctxAB. Our earlier studies showed that variant tcpA (tcpA CIRS and ctxB (ctxB7 first appeared in Kolkata during 2003 and 2006, respectively. The present study revealed that a variant rtxA was first isolated in Kolkata during 2004 and probably formed the genetic background for the emergence of the ctxB7 allele as we were unable to detect a single strain with the combination of El Tor rtxA and ctxB7. The variant gyrA was first time detected in Kolkata during 1994. The Kolkata strains contained four heptad repeats (TTTTGAT in their CT promoter regions whereas Haitian strains carried 5 heptad repeats. Haitian strains had 3 nucleotide deletions at the rstB gene, which is a unique feature of the classical biotype strains. But the Kolkata strains did not have such deletion mutations in the rstB. Our study demonstrated the existence of some Haitian genetic traits in Kolkata isolates along with the dissimilarities in genomic content with respect to rstB and ctxAB promoter region. Finally, we conclude that Haitian variant strain may be evolved due to sequential event in the Indian subcontinent strain with some cryptic modification in the genome.

  13. 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

  14. 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.

  15. Enterotoxigenicity of Mature 45-Kilodalton and Processed 35-Kilodalton Forms of Hemagglutinin Protease Purified from a Cholera Toxin Gene-Negative Vibrio cholerae Non-O1, Non-O139 Strain

    OpenAIRE

    Ghosh, A; Saha, D. R.; Hoque, K. M.; Asakuna, M.; Yamasaki, S.; Koley, H; Das, S. S.; Chakrabarti, M K; Pal, A.

    2006-01-01

    Cholera toxin gene-negative Vibrio cholerae non-O1, non-O139 strain PL-21 is the etiologic agent of cholera-like syndrome. Hemagglutinin protease (HAP) is one of the major secretory proteins of PL-21. The mature 45-kDa and processed 35-kDa forms of HAP were purified in the presence and absence of EDTA from culture supernatants of PL-21. Enterotoxigenicities of both forms of HAP were tested in rabbit ileal loop (RIL), Ussing chamber, and tissue culture assays. The 35-kDa HAP showed hemorrhagic...

  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. Draft Genome Sequence of Environmental Vibrio cholerae 2012EL-1759 with Similarities to the V. cholerae O1 Classical Biotype

    OpenAIRE

    Katz, Lee S.; Turnsek, Maryann; Kahler, Amy; Hill, Vincent R.; Boyd, E. Fidelma; Tarr, Cheryl L.

    2014-01-01

    Vibrio cholerae 2012EL-1759 is an environmental isolate from Haiti that was recovered in 2012 during a cholera outbreak. The genomic backbone is similar to that of the prototypical V. cholerae O1 classical biotype strain O395, and it carries the Vibrio pathogenicity islands (VPI-1 and VPI-2) and a cholera toxin (CTX) prephage.

  18. Vibrio cholerae O1 isolated in Kenya.

    OpenAIRE

    Iwanaga, M; Mori, K.; Kaviti, J N

    1982-01-01

    Biological and serological analyses of 272 isolates of Vibrio cholerae O1 from six epidemics and from a few sporadic cases in Kenya were carried out. All of the isolates were identified as V. cholerae biotype E1 Tor, and 210 out of 272 isolates were hemolytic as examined by Feeley's method.

  19. 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.

  20. The Resistance of Vibrio cholerae O1 El Tor Strains to the Typing Phage 919TP, a Member of K139 Phage Family.

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    Shen, Xiaona; Zhang, Jingyun; Xu, Jialiang; Du, Pengcheng; Pang, Bo; Li, Jie; Kan, Biao

    2016-01-01

    Bacteriophage 919TP is a temperate phage of Vibrio cholerae serogroup O1 El Tor and is used as a subtyping phage in the phage-biotyping scheme in cholera surveillance in China. In this study, sequencing of the 919TP genome showed that it belonged to the Vibrio phage K139 family. The mechanisms conferring resistance to 919TP infection of El Tor strains were explored to help understand the subtyping basis of phage 919TP and mutations related to 919TP resistance. Among the test strains resistant to phage 919TP, most contained the temperate 919TP phage genome, which facilitated superinfection exclusion to 919TP. Our data suggested that this immunity to Vibrio phage 919TP occurred after absorption of the phage onto the bacteria. Other strains contained LPS receptor synthesis gene mutations that disable adsorption of phage 919TP. Several strains resistant to 919TP infection possessed unknown resistance mechanisms, since they did not contain LPS receptor mutations or temperate K139 phage genome. Further research is required to elucidate the phage infection steps involved in the resistance of these strains to phage infection. PMID:27242744

  1. 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.

  2. 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.

    Science.gov (United States)

    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. PMID:27470490

  3. 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. PMID:27487957

  4. Neuraminidase production by Vibrio cholerae O1 and other diarrheagenic bacteria.

    OpenAIRE

    Kabir, S; Ahmad, N.; Ali, S.

    1984-01-01

    Vibrio cholerae O1 strains belonging to both biotypes (classical and El Tor) and both serotypes (Ogawa and Inaba) produced neuraminidase which was released rather than cell bound. Classical strains made more neuraminidase than did El Tor strains. About one-third of V. cholerae non-O1 strains and one-fourth of Aeromonas hydrophila strains were neuraminidase positive. Strains of enterotoxigenic Escherichia coli, Vibrio parahaemolyticus, and Shigella spp. did not produce detectable neuraminidase.

  5. Survival of Vibrio cholerae O1 on fomites.

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    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.

  6. 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. PMID:27430513

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

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    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.

  8. Non-O1, non-O139 Vibrio cholerae bacteraemia in a cirrhotic patient

    OpenAIRE

    Petsaris, O.; Nousbaum, J B; Quilici, M L; Le Coadou, G; PAYAN, C; Abalain, M L

    2010-01-01

    Vibrio cholerae serogroups O1 or O139 are the aetiological agents of cholera. The pathogenicity of non-O1, non-O139 V. cholerae is less well known. These worldwide bacteria are responsible for gastrointestinal infections or, more rarely, bacteraemia in patients with an underlying disease, leading to life-threatening complications. We report a case of non-O1, non-O139 V. cholerae bacteraemia due to a haemolytic strain in a cirrhotic patient. Early antibiotherapy allowed a good outcome. The aim...

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

    OpenAIRE

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

    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.

  10. Worldwide occurrence of integrative conjugative element encoding multidrug resistance determinants in epidemic Vibrio cholerae O1.

    Science.gov (United States)

    Marin, Michel A; Fonseca, Erica L; Andrade, Bruno N; Cabral, Adriana C; Vicente, Ana Carolina P

    2014-01-01

    In the last decades, there has been an increase of cholera epidemics caused by multidrug resistant strains. Particularly, the integrative and conjugative element (ICE) seems to play a major role in the emergence of multidrug resistant Vibrio cholerae. This study fully characterized, by whole genome sequencing, new ICEs carried by multidrug resistant V. cholerae O1 strains from Nigeria (2010) (ICEVchNig1) and Nepal (1994) (ICEVchNep1). The gene content and gene order of these two ICEs are the same, and identical to ICEVchInd5, ICEVchBan5 and ICEVchHai1 previously identified in multidrug resistant V. cholerae O1. This ICE is characterized by dfrA1, sul2, strAB and floR antimicrobial resistance genes, and by unique gene content in HS4 and HS5 ICE regions. Screening for ICEs, in publicly available V. cholerae genomes, revealed the occurrence and widespread distribution of this ICE among V. cholerae O1. Metagenomic analysis found segments of this ICE in marine environments far from the direct influence of the cholera epidemic. Therefore, this study revealed the epidemiology of a spatio-temporal prevalent ICE in V. cholerae O1. Its occurrence and dispersion in V. cholerae O1 strains from different continents throughout more than two decades can be indicative of its role in the fitness of the current pandemic lineage.

  11. 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

  12. Molecular epidemiology of non-O1 Vibrio cholerae and Vibrio mimicus in the U.S. Gulf Coast region.

    OpenAIRE

    Kaper, J B; Nataro, J P; Roberts, N C; Siebeling, R J; Bradford, H B

    1986-01-01

    Ten toxigenic Vibrio cholerae non-O1 and V. mimicus strains isolated from clinical and environmental sources in the U.S. Gulf Coast region were examined for genetic relatedness. Restriction digest patterns of chromosomal DNA and Southern blot analysis with a cholera toxin gene probe revealed that the strains exhibited greater genetic divergence than the highly conserved V. cholerae O1 strains isolated from clinical and sewage samples in this region.

  13. Molecular epidemiology of Vibrio cholerae O1 isolated in Nepal by southern hybridization with a cholera toxin gene probe.

    Science.gov (United States)

    Yamamoto, K; Shrestha, J; Iida, T; Yoh, M; Honda, T

    1995-06-01

    A cholera epidemic broke out in 1992 due to Vibrio cholerae O1 biotype El Tor in the eastern and southern belt of Nepal mainly among the Bhutanese refugees. Restriction fragment profiles (RFP) of DNA fragments of V. cholerae O1 isolates hybridized with an enzyme-labelled oligonucleotide probe for cholera toxin gene (ctx) by Southern Hybridization were compared. The probe hybridized with the 13- and 8-kb fragments of PstI-digested total DNA in all isolates observed in the epidemic. This RFP in the Nepalese strain was not observed in the strains isolated during other epidemics but was observed in the strains isolated from the exported marine products from Taiwan and Thailand. PMID:7594311

  14. 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.

  15. 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.

  16. 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.

  17. Synergistic effect of various virulence factors leading to high toxicity of environmental V. cholerae non-O1/ non-O139 isolates lacking ctx gene : comparative study with clinical strains.

    Directory of Open Access Journals (Sweden)

    Neha Rajpara

    Full Text Available BACKGROUND: Vibrio cholerae non-O1/ non-O139 serogroups have been reported to cause sporadic diarrhoea in humans. Cholera toxins have been mostly implicated for hypersecretion of ions and water into the small intestine. Though most of the V. cholerae non-O1/ non-O139 strains lack these cholera toxins, several other innate virulence factors contribute towards their pathogenicity. The environmental isolates may thus act as reservoirs for potential spreading of these virulence genes in the natural environment which may cause the emergence of epidemic-causing organisms. RESULTS: The environmental isolates of vibrios were obtained from water samples, zooplanktons and phytoplanktons, from a village pond in Gandhinagar, Gujarat, India. They were confirmed as Vibrio cholerae non-O1/ non-O139 using standard biochemical and serotyping tests. PCR experiments revealed that the isolates lacked ctxA, ctxB, tcpA, zot and ace genes whereas other pathogenicity genes like toxR, rtxC, hlyA, hapA and prtV were detected in these isolates. Compared with epidemic strain V. cholerae O1 El Tor N16961, culture supernatants from most of these isolates caused higher cytotoxicity to HT29 cells and higher hemolytic, hemagglutinin and protease activities. In rabbit ileal loop assays, the environmental isolates showed only 2-4 folds lesser fluid accumulation in comparison to N16961 and a V. cholerae clinical isolate IDH02365 of 2009. Pulsed Field Gel electrophoresis and Random amplification of Polymorphic DNA indicated that these isolates showed considerable diversity and did not share the same clonal lineage even though they were derived from the same water source. All the isolates showed resistance to one or more antibiotics. CONCLUSION: Though these environmental isolates lacked the cholera toxins, they seem to have adopted other survival strategies by optimally utilising a diverse array of several other toxins. The current findings indicate the possibility that these

  18. 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

  19. 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

  20. 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

  1. 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.

  2. 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

  3. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  5. Vibrio cholerae non-serogroup O1 cystitis.

    OpenAIRE

    Dumler, J.S.; Osterhout, G J; Spangler, J G; Dick, J D

    1989-01-01

    We report a case of a patient who developed cystitis caused by non-serogroup O1 Vibrio cholerae after swimming in the Chesapeake Bay. Treatment was empirical, with complete symptomatic resolution. Genitourinary tract infections by Vibrio spp. are uncommon but should be considered when cystitis occurs after saltwater exposure in appropriate geographic regions.

  6. 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.

  7. 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.

  8. 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

  9. Characteristics of Vibrio cholerae O1 isolated from water of the River Ganga, Varanasi, India

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    S S Mohaptra

    2015-01-01

    Full Text Available Background: Vibrio cholerae is an autochthonous inhabitant of fresh and brackish water and estuarine system. Investigation of V. cholerae from the River Ganga seems important to find variation in CTX arrangement and genomic diversity. Objectives: To investigate V. cholerae O1 strains for the presence of virulence and regulatory genes, variation in number and organisation of the pre-CTXΦ and/or CTXΦ, and for the genomic diversity. Materials and Methods: Polymerase chain reaction (PCR was used to detect virulence and regulatory genes, type of rstR and location of CTXΦ on the chromosome. Southern hybridisation was conducted to see the number and arrangement of pre-CTXΦ and CTXΦ. Ribotyping and pulsed-field gel electrophoresis were used to find genetic relatedness. Results: Seven strains gave positive results by PCR for the gene encoding for ctx A, zot, ace, tcp A (El Tor, omp U, and tox R, except one strain that was negative for the ctx A. Three strains were positive for the tcp A (El Tor, omp U and tox R genes. Determination of CTX organisation showed that among the ctx-positive strains, four harboured two copies of CTXETΦ arranged in tandem and two harboured one copy of CTXETΦ, and one ctx-negative strain harboured only one copy of pre-CTXETΦ. Pulsotype and ribotype analysis showed existence of at least three pulsotype and ribotypes indicating diversity in genomic content among them. Conclusion: This study thus indicates that multiple clones (ribotypes/pulsotypes of V. cholerae O1 carrying pre-CTXΦ and/or CTXΦ and ctx-negative strains were present in the water of the River Ganga, Varanasi, India.

  10. Genetic Diversity of Vibrio cholerae O1 in Argentina and Emergence of a New Variant

    OpenAIRE

    Pichel, Mariana; Rivas, Marta; Chinen, Isabel; Martín, Fernando; Ibarra, Cristina; Binsztein, Norma

    2003-01-01

    The genetic diversity of Vibrio cholerae O1 strains from Argentina was estimated by random amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE). Twenty-nine isolates carrying the virulence genes ctxA, zot, ace, and tcpA appeared to represent a single clone by both typing methods; while 11 strains lacking these virulence genes exhibited several heterogeneous RAPD and PFGE patterns. Among the last group, a set of isolates from the province Tucumán showed a singl...

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. Attachment of non-culturable toxigenic Vibrio cholerae O1 and non-O1 and Aeromonas spp. to the aquatic arthropod Gerris spinolae and plants in the River Ganga, Varanasi.

    Science.gov (United States)

    Shukla, B N; Singh, D V; Sanyal, S C

    1995-10-01

    Non-cultivable, pathogenic O1 and non-O1 Vibrio cholerae and Aeromonas spp. were resuscitated from aquatic arthropods and plant homogenate respectively, by rabbit ileal loop (RIL) assay. These organisms adhered to the aquatic arthropod Gerris spinolae and various species of phytoplankton in the River Ganga, but failed to grow after direct inoculation on artificial media except for only 10 homogenates of the arthropod. The number of non-O1 V. cholerae and Aeromonas recovered on direct inoculation of G. spinolae homogenates were in the order of 10(5)-10(6) whereas those of the Ganga water were 10(2)-10(3) ml-1. A total of 119 strains of O1 and non-O1 V. cholerae and Aeromonas spp. (69 isolates from G. spinolae and 50 from aquatic plants) were recovered from the loop contents. The results indicate that production of the enzyme chitinase by O1 and non-O1 V. cholerae and Aeromonas spp. might facilitate their adsorption and multiplication on different species of zoo- and phyto-plankton. Most of the isolates were enterotoxic, haemolytic and resistant to different antibiotics. This study suggests that species of zoo- and phyto-planktons, until now not reported to be associated with O1 and non-O1 V. cholerae, may act as reservoirs of these organisms as well as different species of Aeromonas in a fresh-water riverine ecosystem.

  17. 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.

  18. 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.

  19. 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...

  20. 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

  1. 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.

  2. 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.

  3. Vibrio cholerae O1 em amostras de ambientes aquáticos e de alimentos analisados no Estado de Pernambuco, Brasil Isolation of Vibrio cholerae O1 from aquatic environments and foods in Pernambuco State, Brazil

    Directory of Open Access Journals (Sweden)

    Waldêny Colaço

    1998-07-01

    Full Text Available 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 do sorovar Ogawa, foi evidenciada a produção de toxina colérica. A maior incidência do vibrião colérico em águas de rios, canais e de esgoto, representando 86% dos isolados, indicou a contaminação fecal por excretores como a causa preponderante na disseminação da bactéria nos sistemas aquáticos. Assinala-se a discreta ocorrrência de V. cholerae O1 nos alimentos processados (2,1%.Incidence of Vibrio cholerae O1 was studied in 2,585 samples from different aquatic environments and 91 from foods in Pernambuco State, northeastern Brazil, from 1992 to 1994. A total of 193 (7.21% samples of V. cholerae were isolated with a higher prevalence of the Inaba serovar (183-94.8% than the Ogawa serotype (10-5.1%. All isolates were classified as biotype El Tor, and resistance patterns to antibiotics showed that all strains were susceptible tetracycline. Some 70 random samples of Vibrio cholerae proved toxigenic, including all the Ogawa serovars. Incidence of V. cholerae O1 in river water and sewage (86.0% pointed to fecal contamination as the most common source and vehicle for rapid spread of the microorganism in the aquatic environment. The vibrio was isolated in 2.1% of all food examined, which was less than expected.

  4. 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

    2016-02-01

    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. PMID:27180742

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

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    Meer T Alam

    Full Text Available 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.

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

    Science.gov (United States)

    Boucher, Yan

    2016-01-01

    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. PMID:27143391

  7. 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.

  8. Evaluation in mice of a conjugate vaccine for cholera made from Vibrio cholerae O1 (Ogawa O-specific polysaccharide.

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    Mohammad Murshid Alam

    2014-02-01

    Full Text Available BACKGROUND: Protective immunity against cholera is serogroup specific. Serogroup specificity in Vibrio cholerae is determined by the O-specific polysaccharide (OSP of lipopolysaccharide (LPS. Generally, polysaccharides are poorly immunogenic, especially in young children. METHODOLOGY: Here we report the evaluation in mice of a conjugate vaccine for cholera (OSP:TThc made from V. cholerae O1 Ogawa O-Specific Polysaccharide-core (OSP and recombinant tetanus toxoid heavy chain fragment (TThc. We immunized mice intramuscularly on days 0, 21, and 42 with OSP:TThc or OSP only, with or without dmLT, a non-toxigenic immunoadjuvant derived from heat labile toxin of Escherichia coli. PRINCIPAL FINDINGS: We detected significant serum IgG antibody responses targeting OSP following a single immunization in mice receiving OSP:TThc with or without adjuvant. Anti-LPS IgG responses were detected following a second immunization in these cohorts. No anti-OSP or anti-LPS IgG responses were detected at any time in animals receiving un-conjugated OSP with or without immunoadjuvant, and in animals receiving immunoadjuvant alone. Responses were highest following immunization with adjuvant. Serum anti-OSP IgM responses were detected in mice receiving OSP:TThc with or without immunoadjuvant, and in mice receiving unconjugated OSP. Serum anti-LPS IgM and vibriocidal responses were detected in all vaccine cohorts except in mice receiving immunoadjuvant alone. No significant IgA anti-OSP or anti-LPS responses developed in any group. Administration of OSP:TThc and adjuvant also induced memory B cell responses targeting OSP and resulted in 95% protective efficacy in a mouse lethality cholera challenge model. CONCLUSION: We describe a protectively immunogenic cholera conjugate in mice. Development of a cholera conjugate vaccine could assist in inducing long-term protective immunity, especially in young children who respond poorly to polysaccharide antigens.

  9. Unique Clones of Vibrio cholerae O1 El Tor with Haitian Type ctxB Allele Implicated in the Recent Cholera Epidemics from Nigeria, Africa

    Science.gov (United States)

    Pazhani, Gururaja Perumal; Abiodun, Iwalokun Bamidele; Afolabi, Oluwadun; Kolawole, Olukoya Daniel; Mukhopadhyay, Asish K.; Ramamurthy, Thanadarayan

    2016-01-01

    Background and Objectives The antimicrobial susceptibility patterns and genetic characteristics of Vibrio cholerae O1, which is responsible for several cholera epidemics in Nigeria, are not reported in detail since 2007. In this study, we screened V. cholerae O1 El Tor biotype isolates from cholera cases and water samples from different states to investigate their phenotypic and genetic attributes with special reference to their clonality. Results All the V. cholerae O1 biotype El Tor isolates isolated during 2007–2013 were susceptible to fluoroquinolones and tetracycline, the drugs currently used in the treatment of cholera cases in Nigeria. Emergence of CT genotype 7 (Haitian type of ctxB allele) was predominantly seen among Ogawa serotype and the CT genotype 1 (classical ctxB allele) was mostly found in Inaba serotype. Overall, V. cholerae O1 from clinical and water samples were found to be closely related as determined by the pulsed-field gel electrophoresis. V. cholerae isolates from Abia, Kano and Bauchi were found to be genetically distinct from the other states of Nigeria. Conclusion Fecal contamination of the water sources may be the possible source of the cholera infection. Combined prevalence of Haitian and classical ctxB alleles were detected in Ogawa and Inaba serotypes, respectively. This study further demonstrated that V. cholerae O1 with the ctxB has been emerged similar to the isolates reported in Haiti. Our findings suggest that the use of fluoroquinolones or tetracycline/doxycycline may help in the effective management of acute cholera in the affected Nigerian states. In addition, strengthening the existing surveillance in the hospitals of all the states and supply of clean drinking water may control cholera outbreaks in the future. PMID:27479360

  10. 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

  11. 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. PMID:26828665

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

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    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.

  13. 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.

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

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    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.

  15. vttRA and vttRB Encode ToxR Family Proteins That Mediate Bile-Induced Expression of Type Three Secretion System Genes in a Non-O1/Non-O139 Vibrio cholerae Strain▿

    OpenAIRE

    Alam, Ashfaqul; Tam, Vincent; Hamilton, Elaine; Dziejman, Michelle

    2010-01-01

    Strain AM-19226 is a pathogenic non-O1/non-O139 serogroup Vibrio cholerae strain that does not encode the toxin-coregulated pilus or cholera toxin but instead causes disease using a type three secretion system (T3SS). Two genes within the T3SS pathogenicity island, herein named vttRA (locus tag A33_1664) and vttRB (locus tag A33_1675), are predicted to encode proteins that show similarity to the transcriptional regulator ToxR, which is found in all strains of V. cholerae. Strains with a delet...

  16. 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.

  17. 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.

  18. 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. PMID:27222028

  19. New Vibrio cholerae O1 Biotype ElTor bacteriophages

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    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.

  20. 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.

  1. 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. PMID:27255911

  2. 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. PMID:26579088

  3. 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

    Directory of Open Access Journals (Sweden)

    Nadja eBier

    2015-10-01

    Full Text Available 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 towards aminoglycosides. In case of V. cholerae, a noticeable proportion of strains was non-susceptible to aminopenicillins and aminoglycosides. In addition, resistance towards 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.

  4. 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

  5. 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.

  6. Vibrio cholerae non-O1 non-O139 infection in an immunocompromised patient returning from Spain, July 2009.

    Science.gov (United States)

    Rozemeijer, W; Korswagen, L A; Voskuyl, A E; Budding, A E

    2009-01-01

    We describe a severe gastroenteritis with non-O1, non-O139 Vibrio cholerae in an immunocompromised patient returning from a holiday in Spain in July 2009. Predisposing factors and possible cholera enterotoxin production could explain the unusually grave symptomatology. Patient recovered after doxycyclin treatment. PMID:19679033

  7. 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. PMID:27297393

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:26904017

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    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

  13. 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.

  14. 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

  15. 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

  16. 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.

  17. 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. PMID:26881083

  18. 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.

  19. Use of vaginal tampons in sewer surveys for non-O1. Vibrio cholerae.

    OpenAIRE

    Pazzaglia, G; Lesmana, M; Tjaniadi, P; Subekti, D; Kay, B.

    1993-01-01

    Vaginal tampons were shown to be a practical alternative to conventional Moore swabs for isolating Vibrio cholerae from sewage. Associated laboratory investigations demonstrated improved isolation of V. cholerae by using 12- or 18-h enrichments in alkaline peptone water, in comparison with 6-h enrichments, when cultures were incubated at ambient temperatures.

  20. 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.

  1. 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

  2. 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

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

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    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.

  4. Cholera

    Science.gov (United States)

    ... in various settings, either in the form of reactive campaigns in areas experiencing an outbreak or pre- ... to: support the design and implementation of global strategies to contribute to capacity development for cholera prevention ...

  5. Cholera

    Science.gov (United States)

    ... has it), do not offer them. previous continue Prevention Some countries have cholera vaccines that can help ... ON THIS TOPIC Staying Healthy While You Travel Dengue Fever Malaria Typhoid Fever Ebola First Aid: Diarrhea ...

  6. 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).

  7. 甲鱼及养殖水中5株带毒力基因O1群霍乱弧菌的分离与鉴定%Detection and analysis of 5 strains of Vibrio cholerae O1 with virulence gene from green turtles and aquaculture water

    Institute of Scientific and Technical Information of China (English)

    方叶珍; 汪皓秋; 包芳珍; 黄世旺; 邵景莺

    2010-01-01

    目的 监测杭州市江干区水产品中O1群和O139群霍乱弧菌并对分离菌株进行病原学鉴定及分子分型.方法 按第5版对51份样品进行霍乱弧菌分离与鉴定.运用改良Kirby-Bauer纸片法和PCR检测5株霍乱弧菌对14种临床常用抗生素的敏感性及是否携带ctxA和tcpA毒力基因,并用脉冲场凝胶电泳(PFGE)进行分子分型.结果 从51份样品中检出O1群霍乱弧菌5株,其中小川型4株,稻叶型1株.5株霍乱弧菌的ctxA和tcpA毒力基因结果均为阳性.5株菌株对11种抗生素敏感,对红霉素和利福平中介,稻叶型菌株对氨苄西林敏感,小川型菌株对其耐药.PFGE分为两型,两型间仅有一条带的差异.结论 本次分离的5株O1群霍乱弧菌均携带ctxA和tcpA毒力基因,故有必要加强对水产品尤其是甲鱼的监测及管理.

  8. 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.

  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. 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.

  11. 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,

  12. Nontoxigenic Vibrio cholerae Septicemia in an Immunocompromised Patient

    OpenAIRE

    Kamran Kadkhoda; Heather Adam; Gilmour, Matthew W.; Hammond, Gregory W.

    2012-01-01

    We report a recent case of non-O1, non-O139, nontoxigenic Vibrio cholerae septicemia in a post-liver-transplant immunocompromised patient associated with prior seafood consumption. Non-O1, non-O139 V. cholerae strains have been reported in several cases of extraintestinal infections and seem to be emerging infectious agents especially in patients with immunocompromising conditions.

  13. O-Specific Polysaccharide-Specific Memory B Cell Responses in Young Children, Older Children, and Adults Infected with Vibrio cholerae O1 Ogawa in Bangladesh.

    Science.gov (United States)

    Aktar, Amena; Rahman, M Arifur; Afrin, Sadia; Faruk, M Omar; Uddin, Taher; Akter, Aklima; Sami, M Israk Nur; Yasmin, Tahirah; Chowdhury, Fahima; Khan, Ashraful I; Leung, Daniel T; LaRocque, Regina C; Charles, Richelle C; Bhuiyan, Taufiqur Rahman; Mandlik, Anjali; Kelly, Meagan; Kováč, Pavol; Xu, Peng; Calderwood, Stephen B; Harris, Jason B; Qadri, Firdausi; Ryan, Edward T

    2016-05-01

    Cholera caused by Vibrio cholerae O1 confers at least 3 to 10 years of protection against subsequent disease regardless of age, despite a relatively rapid fall in antibody levels in peripheral blood, suggesting that memory B cell responses may play an important role in protection. The V. cholerae O1-specific polysaccharide (OSP) component of lipopolysaccharide (LPS) is responsible for serogroup specificity, and it is unclear if young children are capable of developing memory B cell responses against OSP, a T cell-independent antigen, following cholera. To address this, we assessed OSP-specific memory B cell responses in young children (2 to 5 years, n = 11), older children (6 to 17 years, n = 21), and adults (18 to 55 years, n = 28) with cholera caused by V. cholerae O1 in Dhaka, Bangladesh. We also assessed memory B cell responses against LPS and vibriocidal responses, and plasma antibody responses against OSP, LPS, and cholera toxin B subunit (CtxB; a T cell-dependent antigen) on days 2 and 7, as well as days 30, 90, and 180 after convalescence. In all age cohorts, vibriocidal responses and plasma OSP, LPS, and CtxB-specific responses peaked on day 7 and fell toward baseline over the follow-up period. In comparison, we were able to detect OSP memory B cell responses in all age cohorts of patients with detectable responses over baseline for 90 to 180 days. Our results suggest that OSP-specific memory B cell responses can occur following cholera, even in the youngest children, and may explain in part the age-independent induction of long-term immunity following naturally acquired disease. PMID:27009211

  14. Cholera outbreak caused by drug resistant Vibrio cholerae serogroup O1 biotype ElTor serotype Ogawa in Nepal; a cross-sectional study

    OpenAIRE

    Gupta, Pappu Kumar; Pant, Narayan Dutt; Bhandari, Ramkrishna; Shrestha, Padma

    2016-01-01

    Background Cholera is a major cause of mortality and morbidity in underdeveloped countries including Nepal. Recently drug resistance in Vibrio cholerae has become a serious problem mainly in developing countries. The main objectives of our study were to investigate the occurrence of Vibrio cholerae in stool samples from patients with watery diarrhea and to determine the antimicrobial susceptibility patterns of V. cholerae isolates. Methods A total of 116 stool samples from patients suffering ...

  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. 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

  18. Hybridoma as a specific, sensitive, and ready to use sensing element: a rapid fluorescence assay for detection of Vibrio cholerae O1.

    Science.gov (United States)

    Zamani, Parichehr; Sajedi, Reza H; Hosseinkhani, Saman; Zeinoddini, Mehdi

    2016-09-01

    Over the last decade, isolation and purification of monoclonal antibodies, for diagnostic analysis, have been carried out using the hybridoma expression system. The present study describes a novel example of a detection system using hybridoma cells containing antibody against O1 antigen directly for V. cholerae diagnosis, which is a major health problem in many parts of the world, especially in developing countries. This method has advantages such as simplicity, ease of process, and it does not require manipulation of hybridoma cell. For this approach, an efficient amount of fluorescence calcium indicator, fura 2-AM, was utilized, which emitted light when the intracellular calcium concentration increased as result of antigen binding to specific antibody. More reliable results are obtained via this method and it is considerably faster than other methods, which has the response time of less than 45 s for detection of V. Cholerae O1. Also, the limit of detection was computed to be 50 CFU/mL (cholerae O1. Furthermore, this method was successfully applied to V. cholerae O1 detection in spiked environmental samples, including water and stool samples without any pretreatment. All results reveal that hybridoma cells can provide a valuable, simple, and ready to use tool for rapid detection of other pathogenic bacteria, toxins, and analytes. PMID:27438715

  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 Salmonella Spp., Shigella (Flexneri and Sonnei) and Vibrio Cholerae O1 by Polymerase Chain Reaction (PCR) in Exported Shrimp from the Mexican Northeast Coast

    International Nuclear Information System (INIS)

    The objective of the present work was to use the PCR technique for the simultaneous detection of Salmonella spp and Vibrio cholerae O1 in frozen shrimp for export. The DNA segments located in the gene A [284 pairs of bases (pb)] from Salmonella spp. locus ial (217 and 320 pb) from Shigella flexneri and Shigella sonnei and the gene ctxA and ctxB (777 pb) from Vibrio cholerae O1 were amplified. The different primers that amplify these segments were assayed in a PCR reaction for the simultaneous detection of DNA from the microorganisms. It was not possible to amplify the gene of Shigella sonnei and Shigella flexneri under the assay’s conditions, whilst those of Salmonella spp. and Vibrio cholerae O1 were successfully amplified. The amplification conditions for the PCR were: 94° C, 58° C and 72° C during 30 cycles, allowing a reduction from 15 days test time with the official microbiological methods to 28 hours (24 for the pre-enrichment and four for the PCR). Samples of raw-frozen-headless shrimps were taken from production plants located in the State of Sinaloa, Mexico. A random sampling procedure was used, according to the guidelines described by the International Commission of Microbiological Specifications for Foods (ICMSF, 1999). Five packages per lot per production plant were obtained. From each individual package (5 pounds 80 OZ ≈ 2.27 kg) three samples were taken for the bacteriological assays to search for Salmonella spp. and Vibrio cholerae O1, respectively. The samples were also analyzed by PCR. Results showed that none of the samples were positive by PCR to any of the studied bacteria. Salmonella spp. and Vibrio cholerae O1 were not detected in these samples by the official methods. However, the latter were able to identify other Vibrio species and enterobacteria like Proteus and Acromobacter. These results confirmed PCR’s rapidity, sensitivity and specificity. (author)

  1. 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.

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

    Science.gov (United States)

    Rivard, Nicolas; Ceccarelli, Daniela; Colwell, Rita R.

    2016-01-01

    ABSTRACT 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. PMID:27435459

  3. [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 < 1) indicating the prevalence of the negative V. 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.

  4. Comparison of inferred relatedness based on multilocus variable-number tandem-repeat analysis and whole genome sequencing of Vibrio cholerae O1.

    Science.gov (United States)

    Rashid, Mahamud-Ur; Almeida, Mathieu; Azman, Andrew S; Lindsay, Brianna R; Sack, David A; Colwell, Rita R; Huq, Anwar; Morris, J Glenn; Alam, Munirul; Stine, O Colin

    2016-06-01

    Vibrio cholerae causes cholera, a severe diarrheal disease. Understanding the local genetic diversity and transmission of V. cholerae will improve our ability to control cholera. Vibrio cholerae isolates clustered in genetically related groups (clonal complexes, CC) by multilocus variable tandem-repeat analysis (MLVA) were compared by whole genome sequencing (WGS). Isolates in CC1 had been isolated from two geographical locations. Isolates in a second genetically distinct group, CC2, were isolated only at one location. Using WGS, CC1 isolates from both locations revealed, on average, 43.8 nucleotide differences, while those strains comprising CC2 averaged 19.7 differences. Strains from both MLVA-CCs had an average difference of 106.6. Thus, isolates comprising CC1 were more closely related (P < 10(-6)) to each other than to isolates in CC2. Within a MLVA-CC, after removing all paralogs, alternative alleles were found in all possible combinations on separate chromosomes indicative of recombination within the core genome. Including recombination did not affect the distinctiveness of the MLVA-CCs when measured by WGS. We found that WGS generally reflected the same genetic relatedness of isolates as MLVA, indicating that isolates from the same MLVA-CC shared a more recent common ancestor than isolates from the same location that clustered in a distinct MLVA-CC. PMID:27190166

  5. 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

    OpenAIRE

    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 ...

  6. 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.

  7. Vibrio cholerae non-O1 bacteraemia: description of three cases in the Netherlands and a literature review.

    Science.gov (United States)

    Engel, Madelon F; Muijsken, Mariette A; Mooi-Kokenberg, Esther; Kuijper, Ed J; van Westerloo, David J

    2016-04-14

    Vibrio cholerae non-O1 serogroup (VCNO) bacteraemia is a severe condition with a high case-fatality rate. We report three cases diagnosed in the Netherlands, identified during a national microbiological congress, and provide a literature review on VCNO bacteraemia. A search strategy including synonyms for 'VCNO' and 'bacteraemia' was applied to PubMed, Medline, Web of Science and Embase databases. The three cases were reported in elderly male patients after fish consumption and/or surface water contact. The literature search yielded 82 case reports on 90 cases and six case series. Thirty case reports were from Asia (30/90; 33%), concerned males (67/90; 74%), and around one third (38/90; 42%) involved a history of alcohol abuse and/or liver cirrhosis The presenting symptom often was gastroenteritis (47/90; 52%) which occurred after seafood consumption in 32% of the cases (15/47).Aside from the most frequent symptom being fever, results of case series concurred with these findings. Published cases also included rare presentations e.g. endophthalmitis and neonatal meningitis. Based on the limited data available, cephalosporins seemed the most effective treatment. Although mainly reported in Asia, VCNO bacteraemia occurs worldwide. While some risk factors for VCNO were identified in this study, the source of infection remains often unclear. Clinical presentation may vary greatly and therefore a quick microbiological diagnosis is indispensable. PMID:27104237

  8. 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.

  9. Occurrence, Diversity, and Pathogenicity of Halophilic Vibrio spp. and Non-O1 Vibrio cholerae from Estuarine Waters along the Italian Adriatic Coast

    OpenAIRE

    Barbieri, Elena; Falzano, Loredana; Fiorentini, Carla; Pianetti, Anna; Baffone, Wally; Fabbri, Alessia; Matarrese, Paola; Casiere, Annarita; Katouli, Mohammad; Kühn, Inger; Möllby, Roland; Bruscolini, Francesca; Donelli, Gianfranco

    1999-01-01

    The occurrence, diversity, and pathogenicity of Vibrio spp. were investigated in two estuaries along the Italian Adriatic coast. Vibrio alginolyticus was the predominant species, followed by Vibrio parahaemolyticus, non-O1 Vibrio cholerae, and Vibrio vulnificus. By using a biochemical fingerprinting method, all isolates were grouped into nine phenotypes with similarity levels of 75 to 97.5%. The production of toxins capable of causing cytoskeleton-dependent changes was detected in a large num...

  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. Non-O1 Vibrio cholerae inguinal skin and soft tissue infection with bullous skin lesions in a patient with a penis squamous cell carcinoma

    OpenAIRE

    García-Tutor Emilio; del Pozo Jose L; Yuste Jose R; Portillo María E; Aguinaga Aitziber; Pérez-Gracia Jose L; Leiva José

    2009-01-01

    Abstract Vibrio spp. is a pathogen rarely isolated in cancer patients, and in most cases it is associated with haematological diseases. Cutaneous manifestations of this organism are even rarer. We report a case of Non-O1 Vibrio cholerae inguinal skin and soft tissue infection presenting bullous skin lesions in a young type II diabetic patient with a penis squamous cell carcinoma having a seawater exposure history.

  12. Non-O1 Vibrio cholerae inguinal skin and soft tissue infection with bullous skin lesions in a patient with a penis squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    García-Tutor Emilio

    2009-05-01

    Full Text Available Abstract Vibrio spp. is a pathogen rarely isolated in cancer patients, and in most cases it is associated with haematological diseases. Cutaneous manifestations of this organism are even rarer. We report a case of Non-O1 Vibrio cholerae inguinal skin and soft tissue infection presenting bullous skin lesions in a young type II diabetic patient with a penis squamous cell carcinoma having a seawater exposure history.

  13. 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

  14. 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

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

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    Waldêny Colaço

    1998-07-01

    Full Text Available 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 do sorovar Ogawa, foi evidenciada a produção de toxina colérica. A maior incidência do vibrião colérico em águas de rios, canais e de esgoto, representando 86% dos isolados, indicou a contaminação fecal por excretores como a causa preponderante na disseminação da bactéria nos sistemas aquáticos. Assinala-se a discreta ocorrrência de V. cholerae O1 nos alimentos processados (2,1%.

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

    Directory of Open Access Journals (Sweden)

    Colaço Waldêny

    1998-01-01

    Full Text Available 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 do sorovar Ogawa, foi evidenciada a produção de toxina colérica. A maior incidência do vibrião colérico em águas de rios, canais e de esgoto, representando 86% dos isolados, indicou a contaminação fecal por excretores como a causa preponderante na disseminação da bactéria nos sistemas aquáticos. Assinala-se a discreta ocorrrência de V. cholerae O1 nos alimentos processados (2,1%.

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

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    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

  18. Radiation-sensitive mutant of hypertoxinogenic strain 569B of Vibro cholerae

    International Nuclear Information System (INIS)

    A radiation-sensitive mutant of the hypertoxinogenic strain 569B of Vibrio cholerae was isolated and characterized. The mutant, designated V. cholerae 569Bsub(s), lacks both excision- and medium-dependent dark-repair mechanisms of UV-induced DNA damage while retaining the wild-type photoreactivating capability. Analysis of the UV-irradiated cell DNA by velocity sedimentation in alkaline sucrose gradient suggests that UV-induced pyrimidine dimers may not be incised in these cells. In contrast to the wild-type cells, the mutant cell DNA was degraded after treatment with nalidixic acid. The mutant cells failed to produce any detectable amount of cholera toxin as measured by ileal-loop assay. (orig.)

  19. Radiation-sensitive mutant of hypertoxinogenic strain 569B of Vibro cholerae

    Energy Technology Data Exchange (ETDEWEB)

    Das, G.; Das, J. (Indian Inst. of Chemical Biology, Calcutta. Biophysics Div.)

    1983-04-01

    A radiation-sensitive mutant of the hypertoxinogenic strain 569B of Vibrio cholerae was isolated and characterized. The mutant, designated V. cholerae 569Bsub(s), lacks both excision- and medium-dependent dark-repair mechanisms of UV-induced DNA damage while retaining the wild-type photoreactivating capability. Analysis of the UV-irradiated cell DNA by velocity sedimentation in alkaline sucrose gradient suggests that UV-induced pyrimidine dimers may not be incised in these cells. In contrast to the wild-type cells, the mutant cell DNA was degraded after treatment with nalidixic acid. The mutant cells failed to produce any detectable amount of cholera toxin as measured by ileal-loop assay.

  20. Radiation-sensitive mutant of hypertoxinogenic strain 569B of Vibrio cholerae.

    Science.gov (United States)

    Das, G; Das, J

    1983-04-01

    A radiation-sensitive mutant of the hypertoxinogenic strain 569B of Vibrio cholerae was isolated and characterized. The mutant, designated V. cholerae 569Bs, lacks both excision- and medium-dependent dark-repair mechanisms of UV-induced DNA damage while retaining the wild-type photoreactivating capability. Analysis of the UV-irradiated cell DNA by velocity sedimentation in alkaline sucrose gradient suggests that UV-induced pyrimidine dimers may not be incised in these cells. In contrast to the wild-type cells, the mutant cell DNA was degraded after treatment with nalidixic acid. The mutant cells failed to produce any detectable amount of cholera toxin as measured by ileal-loop assay.

  1. 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

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    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

  2. 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. PMID:24983529

  3. 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

  4. 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. PMID:27077312

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

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    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.

  6. Cholera in Azov area

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    O. N. Domashenko

    2015-07-01

    Full Text Available 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 waters of the city. Typical and watery diarrhoea, vomiting, abdominal pain and lack of normal body temperature, dehydration syndrome, characterized clinical cholera for adults in most cases. The mean duration of diarrhoea was 6,6 days. At 46.9% observed atypical symptoms in 10 (31,3% – abdominal pain (1 patient cramping in 7 cases, localized in the epigastria region, at 2-over stomach. In 5 patients (15,6% had an increase in body temperature to 37,2–37,7 degrees Celsius. In 15 (46,9% patients had severe nausea accompanied by vomiting. Easy for cholera was observed in 1 (3.1%, moderate – in 14 (43,8%, heavy – in 17 (53,1% patients. Dehydration I level is set at 4 (12,5%, II – from 6 (18,7%, III – in 18 (56,3%, IV – 4 (12,5% patients. Cholera outbreak was characterized by a predominance of severe disease and severe dehydration (III and IV, which was observed in 68.8% of patients. The decisive factor in the treatment of cholera patients was initiated in a timely manner rehydration therapy, in particular the introduction of the solution «Trisol». Against the background of rehydration therapy hyperkalaemia was observed in 9,4% of cases, vascular rehydration at 9,4%, the cell rehydration in 3,1% of patients. Fatal accidents cholera outbreaks have not been observed. Conclusion. Clinical diagnosis of cholera and the provision of medical care in the prehospital phase were poor, indicating the need for systematic conducting training seminars among experts

  7. Cholera in 1994. Part I.

    Science.gov (United States)

    1995-07-14

    In 1994, Vibrio cholerae O1 biotype El Tor, the agent responsible for the seventh cholera pandemic which began in 1961, continued to spread in all regions of the world (Map 1). In all, 384,403 cholera cases were officially reported to WHO in 1994 (a 2% increase over 1993), reversing the downward trend which started in 1992. A total of 10,692 deaths were reported in 1994, increasing the reported global case-fatality ratio (CFR) to 2.8% from 1.8% in 1993. Cholera cases were notified from 94 countries/areas, the highest number ever reporting in one year. (Table 1 and Fig. 1). The year was marked by the dramatic cholera outbreak that devastated the Rwandan refugee camps in Goma, Zaire in July. Major outbreaks also affected Afghanistan, Brazil, Guinea, Guinea-Bissau and Somalia. Africa reported a rise in the number of cholera cases, and was the continent accounting for the largest proportion of all reported cases. The incidence of cholera cases in the Americas continued to fall and reported CFR was the lowest recorded since the disease reached Latin America in 1991. Asia reported a 17% increase in cholera cases compared with 1993. Europe, which usually reports only imported cases, registered a significant increase in the number of indigenous cholera cases. Oceania reported 6 cases, 5 of which were imported, and no deaths (Figs. 2 and 3). The new V. cholerae strain O139 (Bengal) has affected 10 countries in Asia since it was first detected in India at PMID:7544150

  8. Comparative and genetic analyses of the putative Vibrio cholerae lipopolysaccharide core oligosaccharide biosynthesis (wav) gene cluster.

    Science.gov (United States)

    Nesper, Jutta; Kraiss, Anita; Schild, Stefan; Blass, Julia; Klose, Karl E; Bockemühl, Jochen; Reidl, Joachim

    2002-05-01

    We identified five different putative wav gene cluster types, which are responsible for the synthesis of the core oligosaccharide (OS) region of Vibrio cholerae lipopolysaccharide. Preliminary evidence that the genes encoded by this cluster are involved in core OS biosynthesis came from analysis of the recently released O1 El Tor V. cholerae genome sequence and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of O1 El Tor mutant strains defective in three genes (waaF, waaL, and wavB). Investigations of 38 different V. cholerae strains by Southern blotting, PCR, and sequencing analyses showed that the O1 El Tor wav gene cluster type is prevalent among clinical isolates of different serogroups associated with cholera and environmental O1 strains. In contrast, we found differences in the wav gene contents of 19 unrelated non-O1, non-O139 environmental and human isolates not associated with cholera. These strains contained four new wav gene cluster types that differ from each other in distinct gene loci, providing evidence for horizontal transfer of wav genes and for limited structural diversity of the core OS among V. cholerae isolates. Our results show genetic diversity in the core OS biosynthesis gene cluster and predominance of the type 1 wav gene locus in strains associated with clinical cholera, suggesting that a specific core OS structure could contribute to V. cholerae virulence.

  9. 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.

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

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    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

  11. 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.

  12. 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.

  13. [Non-toxigenic hemolytic Vibrio cholerae non-O1 non-O139 fatal septicemia. Report of one case].

    Science.gov (United States)

    Briceño L, Isabel; Puebla A, Claudio; Guerra A, Francisco; Jensen F, Daniela; Núñez B, Harold; Ulloa F, María T; Osorio A, Carlos G

    2009-09-01

    We report a 70-year-old woman, who had recently consumed shellfish, that was admitted to the intensive care unit with septic shock and died 19 hours later due to a multi-organic failure. Microbiological, serological and molecular assays confirmed a hemolytic tdh+ Vibrio cholerae non-01, non 0139 as the etiologic agent. PMID:20011960

  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. The El Tor Biotype of Vibrio cholerae Exhibits a Growth Advantage in the Stationary Phase in Mixed Cultures with the Classical Biotype▿

    OpenAIRE

    Pradhan, Subhra; Baidya, Amit K.; Ghosh, Amalendu; Paul, Kalidas; Chowdhury, Rukhsana

    2009-01-01

    Vibrio cholerae strains of the O1 serogroup that typically cause epidemic cholera can be classified into two biotypes, classical and El Tor. The El Tor biotype emerged in 1961 and subsequently displaced the classical biotype as a cause of cholera throughout the world. In this study we demonstrate that when strains of the El Tor and classical biotypes were cocultured in standard LB medium, the El Tor strains clearly had a competitive growth advantage over the classical biotype starting from th...

  16. 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.

  17. 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。

  18. Radiation decontamination of Peruvian marine ''lead snail'' (Thais chocolata) inoculated with Vibrio cholerae O1 El Tor

    International Nuclear Information System (INIS)

    In vivo studies were conducted using marine snails (Thais chocolata) artificially contaminated in a tank containing sea water inoculated with a pure culture of Vibrio cholerae, such that 105 colony forming units per gram (CFU/g) were uptaken by the mollusks in 1.5 h. A radiation D10 value of 0.12 kGy was determined for V. cholerae upon subsequent irradiation of the live snails at doses in the range 0.0-4.0 kGy. A second series of tests were conducted using naturally contaminated, non-inoculated snails, shelled and packaged simulating commercial procedures, irradiated at 0.0-3.0 kGy, and stored at 2-4 deg. C. These tests indicated that a dose of 2.0 kGy was optimal to extend the microbiological shelf-life of the snails to 21 days without inducing significant adverse sensory or chemical effects. Non-irradiated snails similarly treated and stored spoiled after only seven days. (author)

  19. 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.

  20. 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.

  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...

  2. Genome Sequence of the K139-Like Phage VcP032 Originating from the Vibrio cholerae O1 El Tor Ogawa Serotype.

    Science.gov (United States)

    Jäckel, Claudia; Strauch, Eckhard; Hammerl, Jens Andre

    2016-01-01

    Vibrio cholerae is the cause of large cholera outbreaks, especially in endemic regions with high poverty and inadequate sanitation. Here, we announce the complete genome sequence of the virulence-associated broad host range V. cholerae phage VcP032, including a brief summary of its genotypic and phenotypic features. PMID:27445393

  3. Genome Sequence of the K139-Like Phage VcP032 Originating from Vibrio cholerae O1 El Tor Ogawa Serotype

    Science.gov (United States)

    Jäckel, Claudia; Hammerl, Jens Andre

    2016-01-01

    Vibrio cholerae is the cause of large cholera outbreaks, especially in endemic regions with high poverty and inadequate sanitation. Here, we announce the complete genome sequence of the virulence-associated broad host range V. cholerae phage VcP032, including a brief summary of its genotypic and phenotypic features. PMID:27445393

  4. 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...

  5. Study about the sensibility in vitro of different strains of Vibrio cholera 01 exposed to 60 Co gamma radiation

    International Nuclear Information System (INIS)

    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 60 Co. The results are compared with other data related to bacterial food borne diseases, including water. (author)

  6. 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.

  7. 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.

  8. 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

  9. 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

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

    Directory of Open Access Journals (Sweden)

    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

  11. 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.

  12. 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...

  13. 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,...

  14. 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)

  15. 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.

  16. High Prevalence of Vibrio cholerae Non-O1 Carrying Heat-Stable-Enterotoxin-Encoding Genes among Vibrio Isolates from a Temperate-Climate River Basin of Central Italy

    OpenAIRE

    Caldini, G.; Neri, A.; Cresti, S; Boddi, V.; Rossolini, G. M.; Lanciotti, E.

    1997-01-01

    Vibrio spp. of clinical interest from the Arno River basin (Tuscany, Italy) were investigated in this study. Vibrios were isolated from 70% of water samples. Vibrio cholerae non-O1 was the most prevalent species (82% of isolates), followed by Vibrio mimicus (10%) and Vibrio metschnikovii (8%). Recovery of vibrios was correlated with temperature, pH, and various indicators of municipal pollution. None of the 150 Vibrio isolates carried ctx-related genomic sequences, whereas 18 (14.6%) of the 1...

  17. 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

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  20. Critical analysis of compositions and protective efficacies of oral killed cholera vaccines.

    Science.gov (United States)

    Kabir, Shahjahan

    2014-09-01

    Two cholera vaccines, sold as Shanchol and Dukoral, are currently available. This review presents a critical analysis of the protective efficacies of these vaccines. Children under 5 years of age are very vulnerable to cholera and account for the highest incidence of cholera cases and more than half of the resulting deaths. Both Shanchol and Dukoral are two-spaced-dose oral vaccines comprising large numbers of killed cholera bacteria. The former contains Vibrio cholerae O1 and O139 cells, and the latter contains V. cholerae O1 cells with the recombinant B subunit of cholera toxin. In a field trial in Kolkata (India), Shanchol, the preferred vaccine, protected 45% of the test subjects in all of the age groups and only 17% of the children under 5 years of age during the first year of surveillance. In a field trial in Peru, two spaced doses of Dukoral offered negative protection in children under 5 years of age and little protection (15%) in vaccinees over 6 years of age during the first year of surveillance. Little is known about Dukoral's long-term protective efficacy. Both of these vaccines have questionable compositions, using V. cholerae O1 strains isolated in 1947 that have been inactivated by heat and formalin treatments that may denature protein. Immunological studies revealed Dukoral's reduced and short-lived efficacy, as measured by several immunological endpoints. Various factors, such as the necessity for multiple doses, poor protection of children under 5 years of age, the requirement of a cold supply chain, production costs, and complex logistics of vaccine delivery, greatly reduce the suitability of either of these vaccines for endemic or epidemic cholera control in resource-poor settings. PMID:25056361

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  3. 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.

  4. [Antibiotic sensitivity to epidemic strains of Vibrio cholerae and Shigella dysenteriae 1 isolated in Rwandan refugee camps in Zaire].

    Science.gov (United States)

    Cavallo, J D; Niel, L; Talarmin, A; Dubrous, P

    1995-01-01

    Multiresistance or epidemic enteric bacteria to antibiotics greatly complicates treatment, and in some cases prophylaxis, of severe invasive gastroenteritis. During the summer of 1994, two epidemics of diarrhea, one due to Vibrio cholerae and the other to Shigella dysenteriae 1 isolated from the Goma and Bukavu camps was determined by measurement of the Agar Minimal Inhibitory Concentration. Multiresistance to tetracyclins, aminopenicillins, trimethoprimsulfamethoxazole, and nifuroxazide was observed. After intensive treatment mutant forms of both bacteria resistant to nalidixic acid rapidly appeared. Only fluoroquinolones remained active on these mutant strains, but the availability of this agent in Africa is restricted due to cost. The most effective way of preventing resistance is to limit the spread of enteric infections by health education and improvement of hygiene. This can be difficult during wartime. PMID:8830219

  5. Cholera: foodborne transmission and its prevention.

    Science.gov (United States)

    Estrada-García, T; Mintz, E D

    1996-10-01

    The last several years have witnessed a tremendous increase in reported cholera cases across the globe. The explosive arrival of the seventh cholera pandemic in Latin American in 1991, dramatic epidemics of cholera on the Indian subcontinent and in Southeast Asia due to the newly recognized Vibrio cholerae O139 strain, and the often deadly presence of cholera among populations affected by political and social upheaval in Africa and Eastern Europe are evidence that many countries have failed to adopt effective measures for cholera prevention and control. Foodborne transmission of cholera has been well documented by epidemiologic investigations in nearly every continent, and its interruption is a critical component to any integrated programme for cholera prevention and control. We emphasize clear and effective guidelines for the prevention of foodborne cholera transmission that are drawn from a comprehensive review of relevant epidemiologic and laboratory data. PMID:8905306

  6. Regulation of competence-mediated horizontal gene transfer in the natural habitat of Vibrio cholerae.

    Science.gov (United States)

    Metzger, Lisa C; Blokesch, Melanie

    2016-04-01

    The human pathogen Vibrio cholerae is an autochthonous inhabitant of aquatic environments where it often interacts with zooplankton and their chitinous molts. Chitin induces natural competence for transformation in V. cholerae, a key mode of horizontal gene transfer (HGT). Recent comparative genomic analyses were indicative of extensive HGT in this species. However, we can still expand our understanding of the complex regulatory network that drives competence in V. cholerae. Here, we present recent advances, including the elucidation of bipartite competence regulation mediated by QstR, the inclusion of the type VI secretion system in the competence regulon of pandemic O1 El Tor strains, and the identification of TfoS as a transcriptional regulator that links chitin to competence induction in V. cholerae. PMID:26615332

  7. [The role of food in cholera transmission].

    Science.gov (United States)

    Dobosch, D; Gomez Zavaglia, A; Kuljich, A

    1995-01-01

    The spreading of cholera, from Peru to other Latinoamerican countries in 1991, raised questions regarding food safety, food transportation and handling. Control, prevention and risks implied in food import-export were also matters of concern. We deemed it interesting to determine the viability of Vibrio cholerae in wide consumption food locally. Selected food had different intrinsic characteristics such as: acidity (pH), water activity (aw), chemical composition, indigenous flora and other biologic and physic parameters. Twenty food products were contaminated with V. cholerae O1, Ogawa, toxigenic and not toxigenic strains: yoghurt, cream cheese, apricot marmelade, hip rose marmelade, mayonnaise, italian pasta for "empanadas", "dulce de leche", meat sausage, meat and spinach ravioli, margarine, milk dessert (made with cocoa, milk confiture, starch and additives), lettuce, tuna fish, ricotta and sterilized milk. Table I shows the viability of V. cholerae in tested foods, its pH and the reasons why the experiments were ended: 75% of the products studied could tolerate the development of the microorganism for a period ranging from one day (pasta for "empanadas") to ninety days (sterilized milk). Foods with acredity higher than pH 5.5 did not favor the growth of Vibrio. When pH was neutral or slightly acid, viability persisted independently from aw, microbial antagonisms and other physic, chemical or biologic parameters. Nevertheless, other factors such as: surface adherence, amino acids, magnesium and environmental influences not yet well determined, could eventually modify the persistence of V. cholerae in food. According to this study, most food products could tolerate growth and persistence of the infectant agent, up for three months in some cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7565031

  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. Host Response in Rabbits to Infection with Pasteurella multocida Serogroup F Strains Originating from Fowl Cholera

    Science.gov (United States)

    The ability of two avian Pasteurella multocida serogroup F strains to induce disease in rabbits was investigated in this study. Two groups of 18 Pasteurella-free rabbits each were intranasally challenged with strains isolated from chicken and turkey, respectively. Half the animals in each challenge ...

  10. 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.

  11. [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.

  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. 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.

  14. 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

  15. 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

  16. Characterization and Nucleotide Sequence of CARB-6, a New Carbenicillin-Hydrolyzing β-Lactamase from Vibrio cholerae

    OpenAIRE

    Choury, Danièle; Aubert, Gérald; Szajnert, Marie-France; Azibi, Kemal; Delpech, Marc; Paul, Gérard

    1999-01-01

    A clinical strain of Vibrio cholerae non-O1 non-O139 isolated in France produced a new β-lactamase with a pI of 5.35. The purified enzyme, with a molecular mass of 33,000 Da, was characterized. Its kinetic constants show it to be a carbenicillin-hydrolyzing enzyme comparable to the five previously reported CARB β-lactamases and to SAR-1, another carbenicillin-hydrolyzing β-lactamase that has a pI of 4.9 and that is produced by a V. cholerae strain from Tanzania. This β-lactamase is designated...

  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. Vibrio cholerae bacteremia associated with gastrectomy.

    OpenAIRE

    Toeg, A; Berger, S A; Battat, A; Hoffman, M.; Yust, I

    1990-01-01

    Bacteremia due to Vibrio cholerae is rare. Each of 15 cases previously reported in the English language literature occurred in the setting of immune deficiency. We describe an instance of non-serogroup O1 V. cholerae septicemia in an otherwise healthy patient. Susceptibility to such infection may have been enhanced by a prior gastrectomy for duodenal ulcer.

  19. 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.

  20. Emergência da múltipla resistência a antimicrobianos em Vibrio cholerae isolados de pacientes com gastroenterite no Ceará, Brasil Emergence of multiple drug resistance in Vibrio cholerae isolated from patients with gastroenteritis in Ceará, Brazil

    Directory of Open Access Journals (Sweden)

    Ernesto Hofer

    1999-04-01

    Full Text Available Das 7058 amostras de Vibrio cholerae isoladas de pacientes com suspeita de síndrome coleriforme, no período de 1991 a 1993, no Estado do Ceará, foram detectadas duas com as características de múltipla resistência aos antimicrobianos (tetraciclina, ampicilina, eritromicina, sulfametoxazol-trimetoprima e ao composto vibriostático O/129 (2,4-diamino-6,7-diisopropilpteridina. Do ponto de vista bacteriológico uma amostra foi identificada como V. cholerae sorogrupo O:1, biotipo El Tor e sorovar Inaba e a outra, caracterizada como V. cholerae sorogrupo O:22, classificada bioquimicamente no tipo II de Heiberg. Foi demonstrado que apenas na amostra do sorogrupo O:1, a multirresistência era codificada por um plasmídio, transferível por conjugação para Escherichia coli K12 e amostras sensíveis de V. cholerae O1 e não O1, numa freqüência entre 8x10-2 a 5x10-6. O plasmídio responsável pela multirresistência apresentou um peso molecular de 147 Kb, compatível com as descrições em outras partes do mundo.Of 7058 Vibrio cholerae strains recovered from patients suspected of cholera in the State of Ceará between December 1991 and September 1993, two were resistant to antimicrobials (Ampicillin, erythromycin, trimethoprim-sulfamethoxazole, tetracycline and to vibriostatic agent O/129 (2,4-diamino-6,7-diisopropylpteridine. From the bacteriological standpoint, one strain was identified as V. cholerae serogroup O:1, biotype El Tor, serovar Inaba, and another as V. cholerae serogroup O:22, biochemically classified as Heiberg type II. It was shown that only in the serogroup O:1 strain, multiple resistance was encoded by a plasmid transferrable by conjugation to Escherichia coli K12 and a sensitive strains of V. cholerae O1 and non-O1, with at a frequency between 8x10-2 and 5x10-6. The plasmid, with a molecular weight of 147 Kb, encoded both multiple resistance to antimicrobials and the vibriostatic compound (O/129, compatible with descriptions

  1. Shelf-life extension and decontamination of fish fillets (Trachurus picturatus murphyi and Mugil cephalus) and shrimp tails (Penaeus vannamei) inoculated with toxigenic Vibrio cholerae O1 El Tor using gamma radiation

    International Nuclear Information System (INIS)

    The radiation decimal reduction dose (D10) of toxigenic Vibrio cholerae O1 El Tor, Inaba was determined in vitro (0.13 kGy) and in inoculated fresh fillets of saurel (Trachurus picturatus murphyi) (0.12 kGy) and another Pacific fish species known in Peru as ''lisa'', Mugil cephalus (0.13 kGy), both of which are frequently consumed raw in ''ceviche''. The D10 value was similarly determined in tails of the shrimp species Penaeus vannamei (0.13 kGy). In a second phase of the study, radiation doses in the range 1.0-4.0 kGy were evaluated for use in microbiological shelf-life extension of the selected seafood, and for adverse effects on various sensory attributes (appearance, odor, flavor, and texture). A dose of 1.0 kGy doubled the microbiological shelf-life of fish fillets during post-irradiation storage at 0-1 deg. C to approximately 30 days. This dose was deemed optimal also for preserving all sensory characteristics evaluated except appearance, due to a darkening of fillets. Best results in shrimp tails were obtained using 2.0 kGy, which doubled their microbiological shelf-life to 20 days at 0-1 deg. C. Dipping the fillets in a 10% solution of sodium tripolyphosphate before irradiation prevented radiation-induced drip losses. (author)

  2. Inactivation of Vibrio cholerae O1 El Tor inoculated into Peruvian ''choro'' mussels (Aulacomya ater) and two species of clams (Argopecten purpuratus and Gari solida) using medium-dose irradiation

    International Nuclear Information System (INIS)

    The radiation decimal reduction dose (D10) for Vibrio cholerae O1 biotype El Tor inoculated through the natural feeding system into three species of bivalve mollusks from the Peruvian Pacific coast: ''choro'' mussels (Aulacome ater), ''abanico'' clams (Argopecten purpuratus), and common clams (Gari solida), was determined in vivo. The D10 value obtained in vivo was 0.14 kGy in all mollusks tested. Concurrent studies conducted to determine the potential use of irradiation to extend the microbiological shelf-life of the mollusks during post-irradiation storage at 0-1 deg. C indicated that a dose of 1.0 kGy was optimal for choro mussels and abanico clams, whereas 2.0 kGy produced the best results when treating common clams. Shelf-life extension thus achieved was 31 days for choro mussels, 16 days for abanico clams, and 21 days for common clams. Non-irradiated control samples of all mollusks spoiled after 17 days of refrigerated storage. There were no significant (p<.05) adverse effects from the application of the optimal radiation treatments on the sensory characteristics (i.e. appearance, odor, flavor, and texture) of the mollusks. Total volatile basic nitrogen (VBN) and pH values were examined for use as indexes of seafood freshness. (author)

  3. The Vibrio cholerae trh Gene Is Coordinately Regulated In Vitro with Type III Secretion System Genes by VttRA/VttRB but Does Not Contribute to Caco2-BBE Cell Cytotoxicity

    OpenAIRE

    Miller, Kelly A.; Hamilton, Elaine; Dziejman, Michelle

    2012-01-01

    Numerous virulence factors have been associated with pathogenic non-O1/non-O139 serogroup strains of Vibrio cholerae. Among them are the thermostable direct hemolysin (TDH) and the TDH-related hemolysin (TRH), which share amino acid similarities to the TDH and TRH proteins of Vibrio parahaemolyticus, where they have been shown to contribute to pathogenesis. Although TDH and TRH homologs can be encoded on extrachromosomal elements in V. cholerae, type III secretion system (T3SS)-positive strai...

  4. Drinking cholera

    DEFF Research Database (Denmark)

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

    2015-01-01

    Objectives: To measure the salinity levels of common water sources in coastal Bangladesh andexplore perceptions of water palatability among the local population to investigate the plausibility oflinking cholera outbreaks in Bangladesh with ingestion of saline-rich cholera-infected river water. Me...

  5. Phylogeny of Vibrio cholerae Based on recA Sequence

    OpenAIRE

    Stine, O. Colin; Sozhamannan, Shanmuga; Gou, Qing; Zheng, Siqen; Morris, J. Glenn; Johnson, Judith A.

    2000-01-01

    We sequenced a 705-bp fragment of the recA gene from 113 Vibrio cholerae strains and closely related species. One hundred eighty-seven nucleotides were phylogenetically informative, 55 were phylogenetically uninformative, and 463 were invariant. Not unexpectedly, Vibrio parahaemolyticus and Vibrio vulnificus strains formed out-groups; we also identified isolates which resembled V. cholerae biochemically but which did not cluster with V. cholerae. In many instances, V. cholerae serogroup desig...

  6. 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.

  7. 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.

  8. 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.

  9. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. 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.

  12. 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

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

    Science.gov (United States)

    Rebaudet, Stanislas; Mengel, Martin A; Koivogui, Lamine; Moore, Sandra; Mutreja, Ankur; Kande, Yacouba; Yattara, Ousmane; Sarr Keita, Véronique; Njanpop-Lafourcade, Berthe-Marie; Fournier, Pierre-Edouard; Garnotel, Eric; Keita, Sakoba; Piarroux, Renaud

    2014-06-01

    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 disease control

  14. Phage specific for Vibrio cholerae O139 Bengal.

    OpenAIRE

    Albert, M J; Bhuiyan, N A; Rahman, A; Ghosh, A. N.; Hultenby, K; Weintraub, A; Nahar, S; Kibriya, A K; Ansaruzzaman, M; Shimada, T

    1996-01-01

    From the stool of a Vibrio cholerae O139 Bengal-infected patient, a phage that specifically lysed capsulated V. cholerae O139 strains only was isolated. The phage is useful for the confirmatory diagnosis of V. cholerae O139 infection and for the differentiation of variants that lack the capsule.

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Vibrio cholerae hemagglutinin(HA)/protease: An extracellular metalloprotease with multiple pathogenic activities.

    Science.gov (United States)

    Benitez, Jorge A; Silva, Anisia J

    2016-06-01

    Vibrio cholerae of serogroup O1 and O139, the etiological agent of the diarrheal disease cholera, expresses the extracellular Zn-dependent metalloprotease hemagglutinin (HA)/protease also reported as vibriolysin. This enzyme is also produced by non-O1/O139 (non-cholera) strains that cause mild, sporadic illness (i.e. gastroenteritis, wound or ear infections). Orthologs of HA/protease are present in other members of the Vibrionaceae family pathogenic to humans and fish. HA/protease belongs to the M4 neutral peptidase family and displays significant amino acid sequence homology to Pseudomonas aeruginosa elastase (LasB) and Bacillus thermoproteolyticus thermolysin. It exhibits a broad range of potentially pathogenic activities in cell culture and animal models. These activities range from the covalent modification of other toxins, the degradation of the protective mucus barrier and disruption of intestinal tight junctions. Here we review (i) the structure and regulation of HA/protease expression, (ii) its interaction with other toxins and the intestinal mucosa and (iii) discuss the possible role(s) of HA/protease in the pathogenesis of cholera. PMID:26952544

  18. Vibrio Cholerae

    OpenAIRE

    Sri Amelia

    2006-01-01

    Vibrio cholerae adalah salah satu bakteri yang masuk dalam family Vibrionaceae selain dari Aeromonas dan Plesiomonas, dan merupakan bagian dari genus Vibrio. Bakteri ini pertama kali ditemukan oleh Robert Koch pada tahun 1884 dan sangat penting dalam dunia kedokteran karena menyebabkan penyakit kolera, oleh Sri Amelia

  19. 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.

  20. EVALUASI MEDIUM PENGAYAAN Vibrio cholerae UNTUK DIAGNOSIS KOLERA MENGGUNAKAN IMMUNOCHROMATOGRAPHIC STRIP TEST

    Directory of Open Access Journals (Sweden)

    Kambang Sariadji

    2013-05-01

    Full Text Available Abstract. Vibrio cholerae strains are capable of causing  outbreak cholera in developing country with poor sanitation and hygiene .Conventional culture methods currently available for detection of V. cholerae 01 takes 3 – 5 days. Other diagnostic tools are by using rapid immunochromatographic strip test for controlling and preventing the spreading of cholera outbreak. This method has limitation in detection of V.cholerae O1, especially under 105 cfu/mL. Furthermore rapid method can be improved by  enrichment media and incubation in 37° C for 6 – 8 hours. The aims of research are to analyse enrichments media in increasingl V.cholerae O1, and it’s to improve the finding of the laboratory diagnosis of cholera cases. The research was conducted at the Laboratory of Bacteriology, Center of Biomedical and Basic Technology of Health National Institute of Health Research and Development (NIHRD from January - July 2011. Medium evaluation was done by making serial dilutions of Vibrio cholerae O1 from 107-101 cfu / ml  inoculated into three mediums: alkaline peptone water, bismuth sulfite, and gelatin phosphate salt broth medium. Then were incubated 37°C for 8 hours and every two hours was tested by immunochromatographic strip test. The data analysis to determine treatment and individual differences  each group was done by one way ANOVA test. The results showed that alkali peptone water are better than gelatine phosphate salt broth and bismuth sulfite in increasing V.cholerae O1, p.value 0.000 means significant different. Meanwhile from 24 samples dilutions which were inoculated in three enrichment media, and detected by rapid immunochromatographic in every 2 hours for 8 hours showed positive result in enrichments media are 17 samples for alkali peptone water, 13 samples for gelatine phosphate salt broth  and 8 samples for bismuth sulfite. Key Words : V.Cholerae, Enrichment Media, Immunochromatographic strip test   Abstrak. Vibrio cholerae O1

  1. 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

  2. Evaluation of the protective efficacy of Vibrio cholerae ghost (VCG) candidate vaccines in rabbits.

    Science.gov (United States)

    Eko, Francis O; Schukovskaya, Tatiana; Lotzmanova, E Y; Firstova, V V; Emalyanova, N V; Klueva, S N; Kravtzov, A L; Livanova, L F; Kutyrev, Vladimir V; Igietseme, Joseph U; Lubitz, Werner

    2003-09-01

    An effective Vibrio cholerae vaccine is needed to reduce the morbidity and mortality caused by this pathogen. Despite the availability of current oral vaccines with measurable efficacy, there is need for more effective vaccines with broad-spectrum efficacy in target populations. Recent studies have shown that bacterial ghosts, produced by the expression of cloned lysis gene E, possess adjuvant properties and are immunogenic. In this study, ghosts were prepared from V. cholerae O1 or O139 and evaluated as vaccines in the reversible intestinal tie adult rabbit diarrhea (RITARD) model. Rabbits were orally immunized with different doses of V. cholerae ghost (VCG) formulations. The vaccine formulations elicited high levels of serum vibriocidal titers against indicator strains. The magnitude of the response was measured as the geometric mean titer (GMT) increase for all rabbits in relation to prevaccination titers. The induction of cross protection was evidenced by the ability of serum from VCG-immunized rabbits to mediate complement-dependent killing of both the homologous and the heterologous strains. Immunized rabbits were protected against intraduodenal challenge 30 days after primary immunization. Protective immunity against challenge appeared to be dose dependent and was associated with marked inhibition of colonization. These results indicate that VCGs represent a novel approach to cholera vaccine development and constitute an effective vaccine delivery vehicle. PMID:12922096

  3. 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.

  4. 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...

  5. FK phage for differentiating the classical and El T or groups of Vibrio cholerae.

    OpenAIRE

    Takeya, K; Otohuji, T; Tokiwa, H

    1981-01-01

    A new vibrio-infecting phage (FK phage) isolated from sewage lysed all strains of Vibrio cholerae biovar cholerae, whereas all strains of V. cholerae biovar El Tor were resistant to it. FK phage was entirely different from Mukerjee group IV phage in morphology and antigenicity. In addition to group IV phage, the use of FK phage will be useful in the examination and typing of V. cholerae.

  6. CARB-9, a Carbenicillinase Encoded in the VCR Region of Vibrio cholerae Non-O1, Non-O139 Belongs to a Family of Cassette-Encoded β-Lactamases†

    OpenAIRE

    Petroni, Alejandro; Melano, Roberto G.; Saka, Héctor A.; Garutti, Alicia; Mange, Laura; Pasterán, Fernando; Rapoport, Melina; Miranda, Mariana; Faccone, Diego; Rossi, Alicia; Hoffman, Paul S.; Galas, Marcelo F.

    2004-01-01

    The gene blaCARB-9 was located in the Vibrio cholerae super-integron, but in a different location relative to blaCARB-7. CARB-9 (pI 5.2) conferred β-lactam MICs four to eight times lower than those conferred by CARB-7, differing at Ambler's positions V97I, L124F, and T228K. Comparison of the genetic environments of all reported blaCARB genes indicated that the CARB enzymes constitute a family of cassette-encoded β-lactamases.

  7. 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

  8. 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

  9. 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.

  10. 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 str...

  11. 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.

  12. Epidemiology, determinants and dynamics of cholera in pakistan: gaps and prospects for future research

    International Nuclear Information System (INIS)

    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. (author)

  13. 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.

  14. 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. PMID:27247094

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. Polymerase chain reaction for detection of the cholera enterotoxin operon of Vibrio cholerae.

    OpenAIRE

    1991-01-01

    We report a set of oligonucleotide primers and amplification conditions for the polymerase chain reaction to detect the ctx operon of Vibrio cholerae. The results of this assay on strains of V. cholerae and related organisms were identical with those obtained by the DNA colony hybridization test with the polynucleotide probe. The detection limit of this system was 1 pg of chromosomal DNA or broth culture containing three viable cells. The polymerase chain reaction method could directly detect...

  2. 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

  3. 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

  4. 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.

  5. 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.

  6. Cholera risk factors, Papua New Guinea, 2010

    Directory of Open Access Journals (Sweden)

    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.

  7. Vibrio cholerae Biofilms and Cholera Pathogenesis.

    Directory of Open Access Journals (Sweden)

    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.

  8. 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.

  9. 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, ...

  10. Designing an efficient multi-epitope peptide vaccine against Vibrio cholerae via combined immunoinformatics and protein interaction based approaches.

    Science.gov (United States)

    Nezafat, Navid; Karimi, Zeinab; Eslami, Mahboobeh; Mohkam, Milad; Zandian, Sanam; Ghasemi, Younes

    2016-06-01

    Cholera continues to be a major global health concern. Among different Vibrio cholerae strains, only O1 and O139 cause acute diarrheal diseases that are related to epidemic and pandemic outbreaks. The currently available cholera vaccines are mainly lived and attenuated vaccines consisting of V. cholerae virulence factors such as toxin-coregulated pili (TCP), outer membrane proteins (Omps), and nontoxic cholera toxin B subunit (CTB). Nowadays, there is a great interest in designing an efficient epitope vaccine against cholera. Epitope vaccines consisting of immunodominant epitopes and adjuvant molecules enhance the possibility of inciting potent protective immunity. In this study, V. cholerae protective antigens (OmpW, OmpU, TcpA and TcpF) and the CTB, which is broadly used as an immunostimulatory adjuvant, were analyzed using different bioinformatics and immunoinformatics tools. The common regions between promiscuous epitopes, binding to various HLA-II supertype alleles, and B-cell epitopes were defined based upon the aforementioned protective antigens. The ultimately selected epitopes and CTB adjuvant were fused together using proper GPGPG linkers to enhance vaccine immunogenicity. A three-dimensional model of the thus constructed vaccine was generated using I-TASSER. The model was structurally validated using the ProSA-web error-detection software and the Ramachandran plot. The validation results indicated that the initial 3D model needed refinement. Subsequently, a high-quality model obtained after various refinement cycles was used for defining conformational B-cell epitopes. Several linear and conformational B-cell epitopes were determined within the epitope vaccine, suggesting likely antibody triggering features of our designed vaccine. Next, molecular docking was performed between the 3D vaccine model and the tertiary structure of the toll like receptor 2 (TLR2). To gain further insight into the interaction between vaccine and TLR2, molecular dynamics

  11. 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...

  12. 江苏省霍乱病例病原学特征及与外环境污染间的关系%PATHOGENIC CHARACTERISTIC OF CHOLERAE CASES AND ITS CORRELATION WITH ENVIRONMENTAL VIBIRO CHOLERAE POLLUTION IN JIANGSU PROVINCE

    Institute of Scientific and Technical Information of China (English)

    庄菱; 张雪峰; 钱慧敏; 朱凤才; 汤奋扬

    2011-01-01

    [目的]了解江苏省外环境中霍乱弧菌的污染现状,探讨其与病例发生、疾病流行问的关系.[方法]检测水体、水产品和熟食中霍乱弧菌的携带情况,比较外环境来源株与病例株在血清型、耐药性及毒索基因等方面的差异,应用PFGE分型技术分析不同来源菌株的遗传相关性.[结果]株外环境来源株均为不带毒力、对抗生素敏感的O1群非流行株,病人来源株为具有ctxA、zot、ace、tcpA毒力基因的O139群.11株O139群被分成7种PFGE带型,与O1群相似性系数为76.14%.[结论] 外环境来源株在表型及基因水平上与病例株差异较大,不是病例发生的原因.目前霍乱在江苏省处于低频的高度散发状态,近期大规模暴发和流行的可能性较小.%[Objective]To explore the environmental vibrio cholerae pollution situation and its correlation with the occurrence and epidemic of cholera in Jiangsu province.[Methods]Examined vibrio cholerae from water, aquatic products and cooked food.Compared serum type, drug-resistance and virulence genes between strains from environment and patients.Analyzed the genetical correlation among strains from different resources by PFGE.[Results]3 isolates from environment were non-epidemic strains without virulence genes and not sensitive to antibiotics.The strains from patients were 0139 with toxic genes ctxA, zot, ace, tcpA.11 strains of 0139 were classified into 7 PFGE types, and the similarity coefficient was 76.14% with 01 strains.[Conclusion]The strains from environment are different from those from patients on the level of phenotype and gene.The cases are not caused by the strains from environment.At present, the epidemic intensity of cholera in jiangsu province is low and highly sporadic, and the chance of outbreak and prevalent is small.

  13. 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. PMID:27555841

  14. 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. PMID:27555841

  15. 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.

  16. Removal of Cholera Toxin from Aqueous Solution by Probiotic Bacteria

    Directory of Open Access Journals (Sweden)

    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.

  17. 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.

  18. 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.

  19. Third-generation cephalosporin-resistant Vibrio cholerae, India.

    Science.gov (United States)

    Mandal, Jharna; Sangeetha, Vilwanathan; Ganesan, Vithiya; Parveen, Mohamudha; Preethi, Venkatesan; Harish, Belgode Narasimha; Srinivasan, Sampath; Parija, Subhash Chandra

    2012-08-01

    Vibrio cholerae resistance to third-generation cephalosporins is rarely reported. We detected a strain that was negative for extended-spectrum β-lactamase and positive for the AmpC disk test, modified Hodge test, and EDTA disk synergy test and harbored the blaDHA-1 and blaNDM-1 genes. The antimicrobial drug susceptibility profile of V. cholerae should be monitored. PMID:22840562

  20. 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.

  1. 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.

  2. Immunochemical Properties of the Major Outer Membrane Protein of Vibrio cholerae

    OpenAIRE

    Kabir, Shahjahan

    1983-01-01

    Antisera to the major outer membrane protein of Vibrio cholerae (molecular weight, 48,000) raised in rabbits (i) agglutinated several strains of V. cholerae and (ii) immunoprecipitated outer membrane proteins prepared from both the biotypes and serotypes of V. cholerae. Antibodies of all isotypes to the major outer membrane protein were detected in immune human sera by enzyme-linked immunosorbent assay. These results suggest that the major outer membrane protein was the common outer membrane ...

  3. Vibrio cholerae expresses iron-regulated outer membrane proteins in vivo.

    OpenAIRE

    Sciortino, C V; Finkelstein, R A

    1983-01-01

    A comparison was made, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, of the outer membrane proteins of four strains of Vibrio cholerae grown in vivo in infant rabbits and in vitro in low-iron and iron-supplemented defined media. In vivo-grown V. cholerae expressed novel outer membrane-associated proteins which, in part, were similar to those observed on V. cholerae grown in vitro under conditions of iron deprivation.

  4. 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.

  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. 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。

  7. Stabile kloningsvektorer til Vibrio cholerae

    OpenAIRE

    Cohen, Malene; Jakobsen, Henrik; Larsen, Marie B.; Rasmussen, Maria H; Schrøder, Nanna B.

    2005-01-01

    Vi har forsøgt at konstruere stabile, plasmidbaserede ekspressions-/kloningsvektorer til Vibrio cholerae. Ideelt burde sådanne vektorer baseres på naturlige plasmider fra Vibrio cholerae, men idet Vibrio cholerae sjældent indeholder plasmider, tog vi i stedet udgangspunkt i velkarakteriserede kloningsvektorer fra Escherichia coli. I projektet undersøges fire kloningsvektorer med hver deres karakteristika. Det lykkedes at indføre dem i Vibrio cholerae. Med udgangspunkt i den ene vektor vises, ...

  8. Antimicrobial drugs for treating cholera

    OpenAIRE

    Leibovici-Weissman, Ya'ara; Neuberger, Ami; Bitterman, Roni; Sinclair, David; Salam, Mohammed Abdus; Paul, Mical

    2014-01-01

    Background Cholera is an acute watery diarrhoea caused by infection with the bacterium Vibrio cholerae, which if severe can cause rapid dehydration and death. Effective management requires early diagnosis and rehydration using oral rehydration salts or intravenous fluids. In this review, we evaluate the additional benefits of treating cholera with antimicrobial drugs. Objectives To quantify the benefit of antimicrobial treatment for patients with cholera, and determine whether there are diffe...

  9. 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.

  10. 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...

  11. 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.

  12. A localized outbreak of cholera due to vibrio cholerae, ogawa resistant to tetracyclines

    International Nuclear Information System (INIS)

    To study the clinical and laboratory parameters of a localized Cholera outbreak and determine the sensitivity pattern of the subtype involved. Study Design: A descriptive study. Place and Duration of Study: Combined Military Hospital, Lahore. Duration of Study: Two weeks. Patients and Methods: The study is about a localized outbreak of cholera in a group of soldiers, who consumed water from a single contaminated source of water. We are presenting here an account of the clinical and laboratory parameters of 39 hospitalized cases of cholera, who presented with profuse watery diarrhoea and vomiting. There vital signs, hydration status and systemic examination findings were recorded. Stool samples were sent for routine and microscopic examination and bacteriological culture. Blood samples were taken for complete blood count, serum sodium, potassium, urea and creatinine examination. SPSS 18 was used for statistical analysis of the results. Results: The average age of thirty nine men studied in this outbreak was 24.9 ± 6.9 years. There was no statistically significant difference between confirmed and suspected cholera cases on descriptive analysis of the clinical and laboratory parameters. Majority of patients showed pre-renal azotemia which improved within 48 to 72 hours of hospitalization. Stool cultures isolated Vibrio cholerae, subtype Ogawa, which was resistant to tetracyclines, cotrimoxazole and nalidixic acid but sensitive to fluoroquinolones and third generation cephalosporins. The outbreak was controlled when the contaminated water source was sealed and rectified. Conclusion: Multiple drug resistance strains of Vibrio cholera are causing large outbreaks which should be controlled by prevention of the disease and avoiding inappropriate use of antibiotics. (author)

  13. 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.

  14. The core oligosaccharide and thioredoxin of Vibrio cholerae are necessary for binding and propagation of its typing phage VP3.

    Science.gov (United States)

    Zhang, Jingyun; Li, Wei; Zhang, Qian; Wang, Hongxia; Xu, Xiao; Diao, Baowei; Zhang, Lijuan; Kan, Biao

    2009-04-01

    VP3 is a T7-like phage and was used as one of the typing phages in a phage-biotyping scheme that has been used for the typing of Vibrio cholerae O1 biotype El Tor. Here, we studied the receptor and other host genes of V. cholerae necessary for the lytic propagation of VP3. Six mutants resistant to VP3 infection were obtained from the random transposon insertion mutant bank of the sensitive strain N16961. The genes VC0229 and VC0231, which belong to the wav gene cluster encoding the core oligosaccharide (OS) region of lipopolysaccharide, were found to be interrupted by the transposon in five mutants, and the sixth mutant had the transposon inserted between the genes rhlB and trxA, which encode the ATP-dependent RNA helicase RhlB and thioredoxin, respectively. Gene complementation, transcription analysis, and the loss of VP3 sensitivity by the gene deletion mutants confirmed the relationship between VP3 resistance and VC0229, VC0231, and trxA mutation. The product of VP3 gene 44 (gp44) was predicted to be a tail fiber protein. gp44 could bind to the sensitive wild-type strain and the trxA mutant, but not to VC0229 and VC0231 mutants. The results showed that OS is a VP3 receptor on the surface of N16961, thioredoxin of the host strain is involved in the propagation of the phage, and gp44 is the tail fiber protein of VP3. This revealed the first step in the infection mechanism of the T7-like phage VP3 in V. cholerae.

  15. 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.

  16. 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.

  17. 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.

  18. CHOLERA IN BIDAR DISTRICT OF KARNATAKA STATE, INDIA

    Directory of Open Access Journals (Sweden)

    Pramod S

    2014-11-01

    Full Text Available OBJECTIVE: The purpose of this study was to investigate cholera from 2008 August to 2009 August, and create public health importance, in acute diarrhea diseases. Epidemic of cholera have been reported from various parts of our country. (1(2 A total fecal specimen 562 were collected, isolation and identification of pathogen was done according to standard methodology. (3 Antibiotic susceptibility pattern of the isolates were studied, and picked up randomly positive 25 isolates and were sent to national institute of cholera and enteric diseases (NICED Kolkata for confirmation and Phage typing. RESULTS: Of the 562 of patients 57 (10.142% isolates were found to be positive for vibrio. Cholera o1 and non 01/non-0139, out of 25 isolates 19 (76%. The vibrio Cholera 01 ogawa serotype, biotype Eltor and phage type T 27, T 24, T 21, T 13, along with 6 isolates (24% of Non- 0139 was also noticeable as per the report of the (NICED Kolkata. Majority of the clinical isolate were found to be resistant to more than one drug. (1(2

  19. 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

  20. Growth Phase, Oxygen, Temperature, and Starvation Affect the Development of Viable but Non-culturable State of Vibrio cholerae.

    Science.gov (United States)

    Wu, Bin; Liang, Weili; Kan, Biao

    2016-01-01

    Vibrio cholerae can enter into a viable but non-culturable (VBNC) state in order to survive in unfavorable 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 10(6)-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 or 37°C in ASW, most of the cells rapidly died and the culturable cell count reduced from about 10(8) to 10(6)-10(5) 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 roles during the progress of VBNC

  1. 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.

  2. 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.

  3. Genetic mapping of the regulator gene determining enterotoxin synthesis in Vibrio cholerae

    International Nuclear Information System (INIS)

    Data on the genetic mapping of mutation tox-7 (the mutation affecting the synthesis of the cholera toxin) were obtained by conjugation crosses between the atoxigenic donor strain Vibrio cholerae Eltor and the toxigenic recipient strain V. cholera classica. The molecular and genetic analysis of the Tox- recombinants indicated that, when the synthesis of the cholera toxin is disrupted in these strains, the tox-7 mutation (which impairs the regulator gene tox) is gained. Close linkage between the tox-7 and pur-63 mutations was established (during the selection procedure there was 81.1% combined transfer with respect to marker pur-63 situated in the donor strain chromosome more proximal than mutation tox-7). The markers were localized in the following order in the region under investigation: asp-cys-nal-pur-61-trp-his-pur-63-tox-7-ile

  4. 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.

  5. (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.

  6. 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

  7. 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-

  8. 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.

  9. 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.

  10. VIBRIO CHOLERAE 01 CAN ASSUME A "RUGOSE" SURVIVAL FORM THAT RESISTS KILLING BY CHLORINE, YET RETAINS VIRULENCE

    Science.gov (United States)

    V. cholerae 01 are able to shift between smooth and "wrinkled" or "rugose" colonial morphologies. ultures of smooth V. cholerae strains were inactivated in less than 40 and 20 seconds by concentrations of 0.5 mg/L and 1.0 mg/L free chlorine, respectively. n contrast, cultures of ...

  11. 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.

  12. 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。

  13. 2009-2014年梧州市霍乱弧菌监测结果%Surveillance for Vibrio cholerae in Wuzhou city in 2009-2014

    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

  14. A cholera epidemic among the Nicobarese tribe of Nancowry, Andaman, and Nicobar, India.

    Science.gov (United States)

    Sugunan, Attayoor P; Ghosh, Asit R; Roy, Subarna; Gupte, Mohan D; Sehgal, Subhash C

    2004-12-01

    Cholera has not been reported from the Andaman and Nicobar Islands in India. In October 2002, an outbreak of diarrhea occurred among the Nicobarese tribe of the Nancowry group of islands. The outbreak affected 16 of the 45 inhabited villages of three islands with an attack rate of 12.8% and a case fatality ratio of 1.3%. Vibrio cholerae O1 El Tor was isolated from 18 of the 67 patients tested. A study conducted in one of the villages indicated that the outbreak was started there by a person who traveled to a nearby village where an outbreak was occurring. No specific water source could be identified as the source of infection because persons consuming water from all wells were affected. Water samples from 55 sources were tested and 38 of them were contaminated with Escherichia coli. The possible sources of V. cholerae are effluents from ships or poachers from neighboring countries where cholera is endemic. PMID:15642977

  15. Field evaluation of a rapid immunochromatographic dipstick test for the diagnosis of cholera in a high-risk population

    Directory of Open Access Journals (Sweden)

    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.

  16. Virulence factors in environmental and clinical Vibrio cholerae from endemic areas in Kenya

    Directory of Open Access Journals (Sweden)

    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.

  17. 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.

  18. 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.

  19. Genetic Studies of Vibrio cholerae in South West Cameroon—A Phylogenetic Analysis of Isolates from the 2010-2011 Epidemic

    Science.gov (United States)

    Ngwa, Moise C.; Masalla, Thomas; Esemu, Seraphine; Fumoloh, Foche Francis; Kracalik, Ian; Cella, Eleonora; Alam, Meer Taifur; Akoachere, Jane-Francis; Liang, Song; Salemi, Marco; Morris, J. Glenn; Ali, Afsar; Ndip, Lucy M.

    2016-01-01

    Introduction: During the cholera outbreak from 2010 to 2011 in Cameroon, 33,192 cases with 1,440 deaths (case fatality ratio 4.34%) were reported to the World Health Organization. Of these, the South West Region reported 3,120 clinical cases. This region is in the Equatorial Monsoon climatic subzone of Cameroon, close to the coast, raising questions as to whether cases were linked with development of environmental reservoirs. Methods: In an investigation conducted by the Laboratory for Emerging Infectious Diseases, University of Buea, toxigenic V. cholerae O1 were isolated from diarrheal stool samples from 18 patients, with ages ranging from <3 to 70 years. Coordinates for clinical centers at which cases were identified were obtained using a handheld GPS, and were mapped using ArcGIS. Antibiotic susceptibility testing was performed using the Kirby ‘Bauer agar disc diffusion method. The full genomes of these strains were sequenced with the Illumina MiSeq platform. De novo assembly of cholera genomes and multiple sequence alignment were carried out using the bioinformatics pipeline developed in the Emerging Pathogens Institute laboratory at the University of Florida. Results/Discussion: Genetic comparisons showed that isolates were closely related, with pairwise p-distances ranging from 2.25 to 14.52 10-5 nt substitutions per site, and no statistically significant correlation between the pairwise genetic distances and the geographic distances among sampling locations. Indeed, the phylogeny of the Cameroonian strains displays the typical star-like topology and intermixing of strains from different locations that are characteristic of an exponential outbreak localized around a relatively restricted area with occasional spillover to other parts of the country, likely mediated by direct human contact and human movement. Findings highlight the utility of whole genome sequencing and phylogenetic analysis in understanding transmission patterns at the local level. PMID

  20. 广东省2007年霍乱监测的病原特征分析%The etiologic characteristics of Vibrio cholerae in Guangdong province in 2007

    Institute of Scientific and Technical Information of China (English)

    邓小玲; 李柏生; 谭海玲; 孙立梅; 柯碧霞; 柯昌文; 王多春; 阚飙; 钟豪杰

    2008-01-01

    analysis showed that the similarity among Inaba phage 1d strains from different areas were from 94.5% to 100%.However.16 isolates were collected from environment surveillance programs and the predominated biotype could not be found.Additionally,the biotype distribution of cases isolates was not consistent with those isolates through surveillance.High phylogenetic diversity was observed for the same biotypes isolates from cases and surveillance samples.Conclusion Our data showed that V.cholerae O1 El Tor Inaba phage 1d was the predominated biotype with multi-clone coexisting and circulating in Guangdong province in 2007.It also appeared to be the characteristics of cholera in the non-epidemic period,suggesting that it was necessary to enhance the alert surveillance programs for cholera epidemic based on the molecular typing techniques.

  1. 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.

  2. Genome engineering in Vibrio cholerae

    DEFF Research Database (Denmark)

    Val, Marie-Eve; Skovgaard, Ole; Ducos-Galand, Magaly;

    2012-01-01

    Although bacteria with multipartite genomes are prevalent, our knowledge of the mechanisms maintaining their genome is very limited, and much remains to be learned about the structural and functional interrelationships of multiple chromosomes. Owing to its bi-chromosomal genome architecture and its...... 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...

  3. NAD metabolism in Vibrio cholerae.

    OpenAIRE

    Foster, J. W.; Brestel, C

    1982-01-01

    Extracts of Vibrio cholerae were assayed for various enzymatic activities associated with pyridine nucleotide cycle metabolism. The activities measured include NAD glycohydrolase, nicotinamide deamidase, nicotinamide mononucleotide deamidase, and nicotinic acid phosphoribosyltransferase. The results obtained demonstrate the existence in V. cholerae of the five-membered pyridine nucleotide cycle and the potential for a four-membered pyridine nucleotide cycle. The data presented also suggest th...

  4. Sensitivity of Vibrio cholerae cells to lethal and mutagenic effect of UV-irradiation mediated by plasmids

    International Nuclear Information System (INIS)

    The effect of UV-irradiation on Vibrio cholerae cells and its changes mediated by the plasmid R245 have been studied. Vibrio cholerae strains 569B and RV31 have been shown to be considerably more sensitive to lethal effect of UV-irradiation as compared with Escherichia coli and Salmonella typhimurium cells. Highly toxigenic strain 569B and practically atoxigenic strain RV31 have the same UV-sensitivity. Lethla effect of UV-irradiation on Vibrio cholerae cells is incresed when the irradiated cells are plated on enriched media. UV-induction of mutations was not registered in plasmidless strains of Vibrio cholerae. Plasmid R245 increase UV-resistance of vibrio cells and makes them UV-mutable

  5. 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.

  6. EROTYPE IDENTIFICATION OF VIBRIO CHOLERAE BACTERIAWHICH ISOLATED FROM ICE AMONGTUBE AND CUBE ICE TYPE IN FOOD AND BEVERAGES SELLER AT DENPASAR CITY, BALI

    Directory of Open Access Journals (Sweden)

    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.

  7. Water quality in Lake Xochimilco, Mexico: zooplankton indicators and Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Sarma Nandini

    2015-08-01

    Full Text Available Lake Xochimilco is a eutrophic water body in Mexico City used by the local population for aquaculture and agriculture. Water level is maintained with inputs of partially treated waste water from the Cerro de la Estrella treatment plant. In this study we analysed the water quality at two sites of Lake Xochimilco, Lake Xaltocan and the Santa Cruz Canal using various indicators such as zooplankton diversity, saprobic indices, bacterial concentrations and physico-chemical variables. Eighty litres of water was filtered from Lake Xochimilco from each site, once a month from March to October of 2012, and the rotifers, cladocerans and copepods were enumerated and identified. Physico-chemical parameters such as temperature, pH, Secchi depth, water depth, nitrogen and phosphorus and chlorophyll a concentrations, and bacterial densities were measured. During the study we recorded 33 species of rotifers, the most abundant being Brachionus angularis, B. calyciflorus and B. havanaensis.  Among the microcrustaceans the most abundant were the cladoceran Moina micrura and the copepods Acanthocyclops americanus and Arctodiaptomus dorsalis. The species diversity was around 2 bits/ind. and the saprobic index between 1.5-2.0, indicating that both sites were β meso-saprobic.  At both sites nitrogen was <1 mg/L and phosphorus ranged between 2.5-7.8 mg/L. Chlorophyll a concentrations were between 66-136 µg/L. The toxigenic (Vibrio cholerae No-O1/No-O139 and the non-toxigenic (Vibrio cholerae No-O135 strains of the bacterium were recorded, closely associated with littoral rotifers and cladocerans particularly Brachionus quadridentatus and  Alona sp. All variables indicate that these sites in Lake Xochimilco are eutrophic and highly contaminated and that the water quality needs to be improved.

  8. 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.

  9. Adhesion of Vibrio cholerae to Granular Starches

    OpenAIRE

    Gancz, Hanan; Niderman-Meyer, Orly; Broza, Meir; Kashi, Yechezkel; Shimoni, Eyal

    2005-01-01

    Cholera is a severe diarrheal disease caused by specific serogroups of Vibrio cholerae that are pathogenic to humans. Cholera can become epidemic and deadly without adequate medical care. Appropriate rehydration therapy can reduce the mortality rate from as much as 50% of the affected individuals to

  10. A randomized, placebo-controlled trial of the bivalent killed, whole-cell, oral cholera vaccine in adults and children in a cholera endemic area in Kolkata, India.

    Directory of Open Access Journals (Sweden)

    Dilip Mahalanabis

    Full Text Available OBJECTIVES: An effective vaccine against cholera has been used for public health purposes in Vietnam since the 1990s. This vaccine was reformulated to meet WHO requirements. We assessed the safety and immunogenicity of the reformulated bivalent (Vibrio cholerae 01 and 0139 killed whole cell oral vaccine in a cholera endemic area in Kolkata, India. DESIGN: Double-blind, randomized, placebo controlled trial. SETTING: The trial was conducted in the clinical trial ward of the Infectious Diseases Hospital in Kolkata, India. PARTICIPANTS: The participants were 101 healthy adults (males and non-pregnant females aged 18-40 years and 100 healthy children (males and non-pregnant females aged 1-17 years. INTERVENTIONS: Participants were randomized to receive either the bivalent killed whole cell oral cholera vaccine or placebo (killed oral Escherichia coli K12. OUTCOME MEASURES: For safety: proportion of subjects with adverse events during the duration of study participation. For immunogenicity: Proportion of subjects who had a > or = 4-fold rise in serum vibriocidal antibody titers 14 days after the second dose of vaccine or placebo. RESULTS: Adverse reactions were observed with similar frequency among vaccine and placebo recipients in both age groups. Among adults 4% of vaccine and 8% of placebo recipients and among children 4% of vaccine and 2% of placebo recipients had at least one adverse event within 28 days of the first dose of the vaccine. Following immunization, 53% of adult and 80% of children vaccinees showed a > or = 4 fold rise in serum V. cholerae O1 vibriocidal antibody titers. A less pronounced response to V. cholerae O139 vibriocidal antibody titers post-immunization was noted among vaccinees. CONCLUSIONS: We found the vaccine to be safe and immunogenic in a cholera-endemic area in India. TRIAL REGISTRATION: ClinicalTrials.gov NCT00119197.

  11. 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. PMID:27407302

  12. 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)

    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. PMID:21731695

  13. Genome sequencing reveals unique mutations in characteristic metabolic pathways and the transfer of virulence genes between V. mimicus and V. cholerae.

    Directory of Open Access Journals (Sweden)

    Duochun Wang

    Full Text Available 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.

  14. 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.

  15. Lack of Outer Membrane Protein A Enhances the Release of Outer Membrane Vesicles and Survival of Vibrio cholerae and Suppresses Viability of Acanthamoeba castellanii

    Directory of Open Access Journals (Sweden)

    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.

  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. Use of a Cholera Rapid Diagnostic Test during a Mass Vaccination Campaign in Response to an Epidemic in Guinea, 2012

    Science.gov (United States)

    Martinez-Pino, Isabel; Luquero, Francisco J.; Sakoba, Kéïta; Sylla, Souleymane; Haile, Melatwork; Grais, Rebecca F.; Ciglenecki, Iza; Quilici, Marie-Laure; Page, Anne-Laure

    2013-01-01

    Background During the 2012 cholera outbreak in the Republic of Guinea, the Ministry of Health, supported by Médecins Sans Frontières - Operational Center Geneva, used the oral cholera vaccine Shanchol as a part of the emergency response. The rapid diagnostic test (RDT) Crystal VC, widely used during outbreaks, detects lipopolysaccharide antigens of Vibrio cholerae O1 and O139, both included in Shanchol. In the context of reactive use of a whole-cell cholera vaccine in a region where cholera cases have been reported, it is essential to know what proportion of vaccinated individuals would be reactive to the RDT and for how long after vaccination. Methodology/Principal Findings A total of 108 vaccinated individuals, selected systematically among all persons older than one year, were included at vaccination sites and 106 were included in the analysis. Stools samples of this cohort of vaccinated participants were collected and tested with the RDT every day until the test was negative for two consecutive visits or for a maximum of 7 days. A total of 94.3% of cholera vaccine recipients had a positive test after vaccination; all except one of these positive results were reactive only with the O139 antigen. The mean time to become negative in those with an initial positive result after vaccination was 3.8 days, standard deviation 1.1 days. Conclusions/Significance The RDT Crystal VC becomes positive in persons recently vaccinated against cholera, although almost exclusively to the O139 antigen. This reactivity largely disappeared within five days after vaccination. These results suggest that the test can be used normally as soon as 24 hours after vaccination in a context of O1 epidemics, which represent the vast majority of cases, and after a period of five days in areas where V. cholerae O139 is present. The reason why only O139 test line became positive remains to be investigated. PMID:23967359

  19. 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.

  20. In vitro antibacterial activity of onion (allium cepa) against clinical isolates of vibrio cholera

    International Nuclear Information System (INIS)

    Background: Cholera is a major public health problem in developing countries of the world. Bacterial resistance, lack of surveillance data and proper microbiological facilities are major problems regarding diagnosis of cholera. The spread of microbial drug resistance is a global public health challenge that results in increased illness and death rate. Newer antimicrobials or agents are urgently required to overcome this problem. This work was therefore done to investigate the antimicrobial potential of onion against thirty-three clinical isolates of Vibrio cholera. Methods: The extract was prepared by reflux extraction method. Antibacterial screening of clinical isolates of V. cholerae was done by agar well diffusion method. Agar dilution method was used to assess the Minimum Inhibitory Concentration (MIC). Results: All tested strains of V. cholerae were sensitive to onion (Allium cepa) extracts of two types (purple and yellow). Purple type of extract had MIC range of 19.2-21.6 mg/ml. The extract of yellow type onion had an MIC range of 66-68.4 mg/ml. Conclusion: The results indicated that onion (Allium cepa) has an inhibitory effect on V. cholerae. Keeping in view the anti-bacterial activity of this compound can be exploited as a therapeutic agent in an animal model. This finding is a positive point for further investigation of this herb of traditional medicine. (author)

  1. Cholera in the United States

    Centers for Disease Control (CDC) Podcasts

    2011-11-08

    Anna Newton, Surveillance Epidemiologist at CDC, discusses cholera that was brought to the United States during an outbreak in Haiti and the Dominican Republic (Hispaniola).  Created: 11/8/2011 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 11/8/2011.

  2. Maladi Kolera PSA (:60) (Cholera)

    Centers for Disease Control (CDC) Podcasts

    2010-02-18

    This is an important public health announcement about ways you can prevent the spread of cholera. Language: Haitian Creole.  Created: 2/18/2010 by Centers for Disease Control and Prevention (CDC).   Date Released: 2/18/2010.

  3. Of cabbages and chlorine: cholera in Peru.

    Science.gov (United States)

    1992-07-01

    The low case fatality rates (1%) from the 1991 cholera epidemic in Peru was more a result of including diarrheas of a less virulent etiology than that of cholera. In fact, a study during the early phases of the cholera epidemic in Trujillo, Peru revealed that only 79% of suspected cholera cases were infected with vibrio cholera 01. Further other people contended that the government of Peru did not chlorinate many water supplies because studies by the US Environmental Protection Agency suggested that chlorine increases the cancer risk. It reacts with organic matter to make trihalomethanes. 1 study noted that this risk may explain as many as 700 cases of cancer/year in the US, yet cholera was responsible for nearly 40009 deaths in Latin America the 1st year. Besides in Trujillo, Peru the reason for not chlorinating the water supply was not due to a conscious decision to not do so on the part of the government, but because no funds had been made available to purchase chlorinators and chlorine. This is typical of many towns in developing countries. Further raw fish also played a role in transmitting cholera in Peru. Moreover the study in Trujillo indicated that water stored in containers in the home, and not the water supply, was the most important vehicle of transmission. Nevertheless chlorination of both the water supply and stored water would have prevented cholera transmission. In addition, cabbage irrigated with raw wastewater contributed to cholera transmission in Trujillo. But a concern arises if developing countries follow the advice of WHO of 1st treating wastewater in stabilization ponds. Aquatic blue green algae, other zooplankton, and phytoplankton from a microhabitat suitable for V. cholera. In fact, a study in Peru identified a seasonal pattern of the cholera epidemic with the seasonality of V. cholera non-01 from sewage lagoons in Lima. PMID:1351603

  4. Controlling endemic cholera with oral vaccines.

    Directory of Open Access Journals (Sweden)

    Ira M Longini

    2007-11-01

    Full Text Available BACKGROUND: Although advances in rehydration therapy have made cholera a treatable disease with low case-fatality in settings with appropriate medical care, cholera continues to impose considerable mortality in the world's most impoverished populations. Internationally licensed, killed whole-cell based oral cholera vaccines (OCVs have been available for over a decade, but have not been used for the control of cholera. Recently, these vaccines were shown to confer significant levels of herd protection, suggesting that the protective potential of these vaccines has been underestimated and that these vaccines may be highly effective in cholera control when deployed in mass immunization programs. We used a large-scale stochastic simulation model to investigate the possibility of controlling endemic cholera with OCVs. METHODS AND FINDINGS: We construct a large-scale, stochastic cholera transmission model of Matlab, Bangladesh. We find that cholera transmission could be controlled in endemic areas with 50% coverage with OCVs. At this level of coverage, the model predicts that there would be an 89% (95% confidence interval [CI] 72%-98% reduction in cholera cases among the unvaccinated, and a 93% (95% CI 82%-99% reduction overall in the entire population. Even a more modest coverage of 30% would result in a 76% (95% CI 44%-95% reduction in cholera incidence for the population area covered. For populations that have less natural immunity than the population of Matlab, 70% coverage would probably be necessary for cholera control, i.e., an annual incidence rate of < or = 1 case per 1,000 people in the population. CONCLUSIONS: Endemic cholera could be reduced to an annual incidence rate of < or = 1 case per 1,000 people in endemic areas with biennial vaccination with OCVs if coverage could reach 50%-70% depending on the level of prior immunity in the population. These vaccination efforts could be targeted with careful use of ecological data.

  5. Evaluation of a highly discriminating multiplex multi-locus variable-number of tandem-repeats (MLVA) analysis for Vibrio cholerae.

    Science.gov (United States)

    Olsen, Jaran S; Aarskaug, Tone; Skogan, Gunnar; Fykse, Else Marie; Ellingsen, Anette Bauer; Blatny, Janet M

    2009-09-01

    Vibrio cholerae is the etiological agent of cholera and may be used in bioterror actions due to the easiness of its dissemination, and the public fear for acquiring the cholera disease. A simple and highly discriminating method for connecting clinical and environmental isolates of V. cholerae is needed in microbial forensics. Twelve different loci containing variable numbers of tandem-repeats (VNTRs) were evaluated in which six loci were polymorphic. Two multiplex reactions containing PCR primers targeting these six VNTRs resulted in successful DNA amplification of 142 various environmental and clinical V. cholerae isolates. The genetic distribution inside the V. cholerae strain collection was used to evaluate the discriminating power (Simpsons Diversity Index=0.99) of this new MLVA analysis, showing that the assay have a potential to differentiate between various strains, but also to identify those isolates which are collected from a common V. cholerae outbreak. This work has established a rapid and highly discriminating MLVA assay useful for track back analyses and/or forensic studies of V. cholerae infections. PMID:19555725

  6. Toxin-coregulated pilus-loaded microparticles as a vaccine against Vibrio cholerae O139

    Institute of Scientific and Technical Information of China (English)

    杜艳; 贾文祥; 刘莉

    2004-01-01

    @@ The cholera epidemics is an important public health problem in many developing countries. Highly effective and preventive vaccines against cholera are under investigation as alternatives to the one available presently. Much of the vaccine research focuses on colonization factors. Colonization of a human by the Vibrio cholerae (V. cholerae Ol strain is mediated by toxin-coregulated pilus (TCP), 1 which was shown to play a role in the infant mouse cholera model and subsequently in human volunteers. 2 TCP-loaded vaccines could potentially provide cross-protection among experimental strains. Data have indicated that poly (D,L-lactide)-polyethylene glycol copolymer (PELA)microparticles loaded antigens were strongly immunogenic, 3 and that these microparticles served as an effective delivery system for a single dose of vaccine. 4Microparticle formulation could represent the next generation of vaccines, as they are highly effective at delivery of vaccines, thus requiring fewer doses. 5 We prepared PELA microparticles loaded with TCP for testing as a vaccine; their targeting distributions were identified and related immune responses were analyzed.

  7. Characterization of Vibrio cholerae bacteriophages isolated from the environmental waters of the Lake Victoria region of Kenya.

    Science.gov (United States)

    Maina, Alice Nyambura; Mwaura, Francis B; Oyugi, Julius; Goulding, David; Toribio, Ana L; Kariuki, Samuel

    2014-01-01

    Over the last decade, cholera outbreaks have become common in some parts of Kenya. The most recent cholera outbreak occurred in Coastal and Lake Victoria region during January 2009 and May 2010, where a total of 11,769 cases and 274 deaths were reported by the Ministry of Public Health and Sanitation. The objective of this study is to isolate Vibrio cholerae bacteriophages from the environmental waters of the Lake Victoria region of Kenya with potential for use as a biocontrol for cholera outbreaks. Water samples from wells, ponds, sewage effluent, boreholes, rivers, and lakes of the Lake Victoria region of Kenya were enriched for 48 h at 37 °C in broth containing a an environmental strain of V. cholerae. Bacteriophages were isolated from 5 out of the 42 environmental water samples taken. Isolated phages produced tiny, round, and clear plaques suggesting that these phages were lytic to V. cholerae. Transmission electron microscope examination revealed that all the nine phages belonged to the family Myoviridae, with typical icosahedral heads, long contractile tails, and fibers. Head had an average diameter of 88.3 nm and tail of length and width 84.9 and 16.1 nm, respectively. Vibriophages isolated from the Lake Victoria region of Kenya have been characterized and the isolated phages may have a potential to be used as antibacterial agents to control pathogenic V. cholerae bacteria in water reservoirs.

  8. 霍乱弧菌及其它致病弧菌分子遗传特征和DNA多态性研究%The inherent characteristics and DNA polymorphism of Vibrio cholerae and other vibrios

    Institute of Scientific and Technical Information of China (English)

    王军; 李跃旗; 石建时; 李立新; 白文林; 虞爱华; 姜素椿

    2002-01-01

    Objective To investigate the inherent characteristics of Vibrio cholerae (V. cholerae) and other vibrios and their relationship. Methods Polymerase chain reaction (PCR), DNA sequence analysis, randomly amplified polymorphic DNA (RAPD) analysis and average linkage cluster analysis were used to study 3 isolates of V. cholerae strains O139, three isolates O1 biotype El Tor, four isolates O1 biotype classical and 3 other vibrios.Conclusion V. cholerae and other vibrios are polymorphic in inherent characteristics. The inherent characteristics of V. cholerae O139 are the same as El Tor biotype. O139 may have evolved from the El Tor biotype. The inherent characteristics of vibrio paraheamolyticus are the same as vibrio vulnificus.%目的探讨霍乱弧菌和其它致病弧菌分子遗传特征及相互关系.方法用聚合酶链反应(Polymerase chain reaction; PCR)、DNA 序列分析、随机扩增多态性 DNA (Randomly amplified polymorphic DNA; RAPD)和平均链锁聚类分析,对3株O139群霍乱弧菌、3株O1群 El Tor型、4株古典型和3株副溶血等致病弧菌进行检测.结果 O139群霍乱弧菌和O1群含有相同的霍乱肠毒素(CTX)A2-B亚单位基因, 核苷酸序列同源性为97.1%-98.9%.RAPD 将不同弧菌分成4类;即①O139群和El Tor型、②古典型、③副溶血和创伤弧菌及④河弧菌.O139群与O1群El Tor型DNA指纹图谱几乎完全一致,平均链锁距离为0,与古典型相似,平均链锁距离为2.07. 与副溶血等致病弧菌差别较大,平均链锁距离为6.76-8.54.结论 O139群霍乱弧菌与O1群El Tor型遗传特征相同,前者很可能由El Tor型进化而来.副溶血与创伤弧菌也具有相同遗传特征.霍乱弧菌和其它致病弧菌遗传特征表现出多态性.

  9. Single Nucleotide Polymorphisms in Regulator-Encoding Genes Have an Additive Effect on Virulence Gene Expression in a Vibrio cholerae Clinical Isolate

    Science.gov (United States)

    Carignan, Bailey M.; Brumfield, Kyle D.

    2016-01-01

    ABSTRACT Vibrio cholerae is the etiological agent of the infectious disease cholera, which is characterized by vomiting and severe watery diarrhea. Recently, V. cholerae clinical isolates have demonstrated increased virulence capabilities, causing more severe symptoms with a much higher rate of disease progression than previously observed. We have identified single nucleotide polymorphisms (SNPs) in four virulence-regulatory genes (hapR, hns, luxO, and vieA) of a hypervirulent V. cholerae clinical isolate, MQ1795. Herein, all SNPs and SNP combinations of interest were introduced into the prototypical El Tor reference strain N16961, and the effects on the production of numerous virulence-related factors, including cholera toxin (CT), the toxin-coregulated pilus (TCP), and ToxT, were analyzed. Our data show that triple-SNP (hapR hns luxO and hns luxO vieA) and quadruple-SNP combinations produced the greatest increases in CT, TCP, and ToxT production. The hns and hns luxO SNP combinations were sufficient for increased TCP and ToxT production. Notably, the hns luxO vieA triple-SNP combination strain produced TCP and ToxT levels similar to those of MQ1795. Certain SNP combinations (hapR and hapR vieA) had the opposite effect on CT, TCP, and ToxT expression. Interestingly, the hns vieA double-SNP combination strain increased TCP production while decreasing CT production. Our findings suggest that SNPs identified in the four regulatory genes, in various combinations, are associated with increased virulence capabilities observed in V. cholerae clinical isolates. These studies provide insight into the evolution of highly virulent strains. IMPORTANCE Cholera, an infectious disease of the small intestine caused by the aquatic bacterium Vibrio cholerae, often results in vomiting and acute watery diarrhea. If left untreated or if the response is too slow, the symptoms can quickly lead to extreme dehydration and ultimately death of the patient. Recent anecdotal evidence of

  10. Comparative genomic analysis shows that avian pathogenic Escherichia coli isolate IMT5155 (O2:K1:H5; ST complex 95, ST140 shares close relationship with ST95 APEC O1:K1 and human ExPEC O18:K1 strains.

    Directory of Open Access Journals (Sweden)

    Xiangkai Zhu Ge

    Full Text Available Avian pathogenic E. coli and human extraintestinal pathogenic E. coli serotypes O1, O2 and O18 strains isolated from different hosts are generally located in phylogroup B2 and ST complex 95, and they share similar genetic characteristics and pathogenicity, with no or minimal host specificity. They are popular objects for the study of ExPEC genetic characteristics and pathogenesis in recent years. Here, we investigated the evolution and genetic blueprint of APEC pathotype by performing phylogenetic and comparative genome analysis of avian pathogenic E. coli strain IMT5155 (O2:K1:H5; ST complex 95, ST140 with other E. coli pathotypes. Phylogeny analyses indicated that IMT5155 has closest evolutionary relationship with APEC O1, IHE3034, and UTI89. Comparative genomic analysis showed that IMT5155 and APEC O1 shared significant genetic overlap/similarities with human ExPEC dominant O18:K1 strains (IHE3034 and UTI89. Furthermore, the unique PAI I5155 (GI-12 was identified and found to be conserved in APEC O2 serotype isolates. GI-7 and GI-16 encoding two typical T6SSs in IMT5155 might be useful markers for the identification of ExPEC dominant serotypes (O1, O2, and O18 strains. IMT5155 contained a ColV plasmid p1ColV5155, which defined the APEC pathotype. The distribution analysis of 10 sequenced ExPEC pan-genome virulence factors among 47 sequenced E. coli strains provided meaningful information for B2 APEC/ExPEC-specific virulence factors, including several adhesins, invasins, toxins, iron acquisition systems, and so on. The pathogenicity tests of IMT5155 and other APEC O1:K1 and O2:K1 serotypes strains (isolated in China through four animal models showed that they were highly virulent for avian colisepticemia and able to cause septicemia and meningitis in neonatal rats, suggesting zoonotic potential of these APEC O1:K1 and O2:K1 isolates.

  11. Comparative genomics of 274 Vibrio cholerae genomes reveals mobile functions structuring three niche dimensions

    NARCIS (Netherlands)

    Dutilh, B.E.; Thompson, C.C.; Vicente, A.C.; Marin, M.A.; Lee, C.; Silva, G.G.; Schmieder, R.; Andrade, B.G.; Chimetto, L.; Cuevas, D.; Garza, D.R.; Okeke, I.N.; Aboderin, A.O.; Spangler, J.; Ross, T.; Dinsdale, E.A.; Thompson, F.L.; Harkins, T.T.; Edwards, R.A.

    2014-01-01

    BACKGROUND: Vibrio cholerae is a globally dispersed pathogen that has evolved with humans for centuries, but also includes non-pathogenic environmental strains. Here, we identify the genomic variability underlying this remarkable persistence across the three major niche dimensions space, time, and h

  12. Assessing clonal correlation of epidemic Vibrio cholerae isolates during 2011 in 16 provinces of Iran.

    Science.gov (United States)

    Hajia, Massoud; Rahbar, Mohamad; Farzami, Marjan Rahnamye; Asl, Hossein Masoumi; Dolatyar, Alireza; Imani, Mohsen; Saburian, Roghieh; Mafi, Moharam; Bakhshi, Bita

    2015-03-01

    A total of 1,187 Vibrio cholerae isolates were received during 2011 cholera outbreaks from 16 provinces in different geographical location to Iranian reference Health laboratory. A random selection was performed, and 61 isolates were subjected to further investigations. Cholera cases were come up from May with nine cases and reached to its maximum rate at August (57 cases) and continued to October after which a fall occurred in September. All of the isolates were susceptible to three antimicrobial agents including ciprofloxacin, cefixime, and ampicillin. The highest rate of resistance was seen to nalidixic acid (96.7 %) and co-trimoxazole (91.8 %). Clonality of isolates was investigated through genotyping by PFGE method. A total of seven pulsotypes were obtained from 61 isolates under study. The pulsotypes were highly related with only 1-3 bands differences. Three pulsotypes (PT5, PT6, and PT7) constituted 93.4 % of total isolates. One environmentally isolated strain showed distinct pattern from clinical specimens. This strain although had no any evidence in identified cholera infections, highlighted selecting more environmental specimens in any future outbreaks as long as human samples. In conclusion, emergence and dominance of Ogawa serotypes after about 7 years in Iran are alarming due to fear of import of new V. cholerae clones from out of the country. Approximately, one third of patients in 2011 cholera outbreak in Iran were of Afghan or Pakistani nationality which makes the hypothesis of import of Ogawa serotype strains from neighboring countries more documented and signifies the need to monitor and protect the boundaries.

  13. The Type II secretion system delivers matrix proteins for biofilm formation by Vibrio cholerae.

    Science.gov (United States)

    Johnson, Tanya L; Fong, Jiunn C; Rule, Chelsea; Rogers, Andrew; Yildiz, Fitnat H; Sandkvist, Maria

    2014-12-01

    Gram-negative bacteria have evolved several highly dedicated pathways for extracellular protein secretion, including the type II secretion (T2S) system. Since substrates secreted via the T2S system include both virulence factors and degradative enzymes, this secretion system is considered a major survival mechanism for pathogenic and environmental species. Previous analyses revealed that the T2S system mediates the export of ≥ 20 proteins in Vibrio cholerae, a human pathogen that is indigenous to the marine environment. Here we demonstrate a new role in biofilm formation for the V. cholerae T2S system, since wild-type V. cholerae was found to secrete the biofilm matrix proteins RbmC, RbmA, and Bap1 into the culture supernatant, while an isogenic T2S mutant could not. In agreement with this finding, the level of biofilm formation in a static microtiter assay was diminished in T2S mutants. Moreover, inactivation of the T2S system in a rugose V. cholerae strain prevented the development of colony corrugation and pellicle formation at the air-liquid interface. In contrast, extracellular secretion of the exopolysaccharide VPS, an essential component of the biofilm matrix, remained unaffected in the T2S mutants. Our results indicate that the T2S system provides a mechanism for the delivery of extracellular matrix proteins known to be important for biofilm formation by V. cholerae. Because the T2S system contributes to the pathogenicity of V. cholerae by secreting proteins such as cholera toxin and biofilm matrix proteins, elucidation of the molecular mechanism of T2S has the potential to lead to the development of novel preventions and therapies. PMID:25266381

  14. [Various aspects of the seventh cholera pandemic].

    Science.gov (United States)

    Ladnyĭ, I D

    1976-07-01

    Problems on epidemiological surveillance over cholera are elucidated; recommendations elaborated at the meeting in Madrid in 1975 are given. Data on cholera morbidity on different continent in 1961-1975, with indication of the number of countries which sent their reports to the WHO, are presented. PMID:1007738

  15. Repair of ultraviolet light-induced DNA damage in cholera bacteriophages

    Energy Technology Data Exchange (ETDEWEB)

    Palit, B.N.; Das, G.; Das, J. (Indian Inst. of Chemical Biology, Calcutta. Dept. of Biophysics)

    1983-08-01

    DNA repair-proficient and -deficient strains of Vibrio cholerae were used to examine host cell reactivation, Weigle reactivation and photoreactivation of u.v.-irradiated cholera bacteriophages. U.v. light-induced DNA damage in phages of different morphological and serological groups could be efficiently photoreactivated. Host cell reactivation of irradiated phages of different groups was different on the same indicator host. Phage phi149 was the most sensitive, and phi138 the most resistant to u.v. irradiation. While phi138 showed appreciable host cell reactivation, this was minimal for phi149. Attempts to demonstrate Weigle reactivation of u.v.-irradiated cholera phages were not successful, although u.v.-induced filamentation of host cells was observed.

  16. Repair of ultraviolet light-induced DNA damage in cholera bacteriophages

    International Nuclear Information System (INIS)

    DNA repair-proficient and -deficient strains of Vibrio cholerae were used to examine host cell reactivation, Weigle reactivation and photoreactivation of u.v.-irradiated cholera bacteriophages. U.v. light-induced DNA damage in phages of different morphological and serological groups could be efficiently photoreactivated. Host cell reactivation of irradiated phages of different groups was different on the same indicator host. Phage phi149 was the most sensitive, and phi138 the most resistant to u.v. irradiation. While phi138 showed appreciable host cell reactivation, this was minimal for phi149. Attempts to demonstrate Weigle reactivation of u.v.-irradiated cholera phages were not successful, although u.v.-induced filamentation of host cells was observed. (author)

  17. Why treatment centres failed to prevent cholera deaths among Rwandan refugees in Goma, Zaire.

    Science.gov (United States)

    Siddique, A K; Salam, A; Islam, M S; Akram, K; Majumdar, R N; Zaman, K; Fronczak, N; Laston, S

    1995-02-11

    In July, 1994, in one of the worst cholera outbreaks in recent times, an estimated 12,000 Rwandan refugees died in Goma in eastern Zaire. The Vibrio cholerae strains were resistant to tetracycline and doxycycline, the commonly used drugs for cholera treatment. Despite the efforts of international organisations, which provided medical relief by establishing treatment centres in Goma, mortality from the disease was much higher than expected. In the area of Muganga camp, which had the largest concentration of refugees and where most of the medical aid organisations were active, the highest reported case-fatality ratio for a single day was 48%. The slow rate of rehydration, inadequate use of oral rehydration therapy, use of inappropriate intravenous fluids, and inadequate experience of health workers in management of severe cholera are thought to be some of the factors associated with the failure to prevent so many deaths during the epidemic. In one of the temporary treatment centres with the worst case-fatality record, our team showed that improvement of these factors could increase the odds of survival of cholera patients even in a disaster setting.

  18. Extracts of edible and medicinal plants damage membranes of Vibrio cholerae.

    Science.gov (United States)

    Sánchez, Eduardo; García, Santos; Heredia, Norma

    2010-10-01

    The use of natural compounds from plants can provide an alternative approach against food-borne pathogens. The mechanisms of action of most plant extracts with antimicrobial activity have been poorly studied. In this work, changes in membrane integrity, membrane potential, internal pH (pH(in)), and ATP synthesis were measured in Vibrio cholerae cells after exposure to extracts of edible and medicinal plants. A preliminary screen of methanolic, ethanolic, and aqueous extracts of medicinal and edible plants was performed. Minimal bactericidal concentrations (MBCs) were measured for extracts showing high antimicrobial activity. Our results indicate that methanolic extracts of basil (Ocimum basilicum L.), nopal cactus (Opuntia ficus-indica var. Villanueva L.), sweet acacia (Acacia farnesiana L.), and white sagebrush (Artemisia ludoviciana Nutt.) are the most active against V. cholera, with MBCs ranging from 0.5 to 3.0 mg/ml. Using four fluorogenic techniques, we studied the membrane integrity of V. cholerae cells after exposure to these four extracts. Extracts from these plants were able to disrupt the cell membranes of V. cholerae cells, causing increased membrane permeability, a clear decrease in cytoplasmic pH, cell membrane hyperpolarization, and a decrease in cellular ATP concentration in all strains tested. These four plant extracts could be studied as future alternatives to control V. cholerae contamination in foods and the diseases associated with this microorganism. PMID:20802077

  19. Specific Detection of Toxigenic Vibrio cholerae Based on in situ PCR in Combination With Flow Cytometry

    Institute of Scientific and Technical Information of China (English)

    LI ZHU; JUN-PENG CAI; QING CHEN; SHOU-YI YU

    2007-01-01

    Objective To develop an in situ PCR in combination with flow cytometry (ISPCR-FCM) for monitoring cholera toxin positive Vibrio cholerae. Methods In running this method, 4% paraformaldehyde was used to fix the Vibrio cholerae cells and 1 mg/mL lysozyme for 20 min to permeabilize the cells. Before the PCR thermal cycling, 2.5% glycerol was added into the PCR reaction mixture in order to protect the integrality of the cells. Results A length of 1037bp DNA sequence was amplified, which is specific for the cholera toxin gene (ctxAB gene). Cells subjected to ISPCR showed the presences of ctxAB gene both in epifiuorescence microscopy and in flow cytometric analysis. The specificity and sensitivity of the method were investigated. The sensitivity was relatively low (105 cells/mL), while the specificity was high. Conclusion We have successfully developed a new technique for detection of toxigenic Vibrio cholerae strains. Further study is needed to enhance its sensitivities. ISPCR-FCM shows a great promise in monitoring specific bacteria and their physiological states in environmental samples.

  20. Vibrio cholerae as a predator: lessons from evolutionary principles

    OpenAIRE

    StefanPukatzki; DanieleProvenzano

    2013-01-01

    Diarrheal diseases are the second-most common cause of death among children under the age of five worldwide. Cholera alone, caused by the marine bacterium Vibrio cholerae, is responsible for several million cases and over 120,000 deaths annually. When contaminated water is ingested, V. cholerae passes through the gastric acid barrier, penetrates the mucin layer of the small intestine, and adheres to the underlying epithelial lining. V. cholerae multiplies rapidly, secretes cholera toxin, and ...

  1. Investigation of water sources as reservoirs of Vibrio cholerae in Bepanda, Douala and determination of physico-chemical factors maintaining its endemicity

    Directory of Open Access Journals (Sweden)

    Akoachere J.-F.K. Tatah

    2012-06-01

    Full Text Available Cholera remains a significant cause of mortality in developing countries. Outbreaks of the disease are associated with poverty, lack of potable water and poor sanitation. The survival and persistence of Vibrio cholerae in water has been shown to depend on physico-chemical factors. We studied water sources in Bepanda, an overcrowded neighbourhood in Douala, Cameroon, with limited access to portable water and very poor sanitary conditions as reservoirs of V. cholerae. We analysed 318 samples from various sources (well, tap, stream from February to July 2009 using standard microbiological techniques and characterised isolates serologically using the polyvalent O1/O139 antisera. Susceptibility to antibiotics previously used for cholera treatment in Douala was studied using the disk diffusion method. Physico-chemical factors (temperature, pH and salinity that could maintain the endemicity of the organism were analysed using standard methods. Eighty-seven (27.4% samples were contaminated, with high isolation rates being obtained from streams (52.4% and wells (29.8%. The number of isolates was significantly higher (P < 0.05 in the rainy season (35.5%. We detected 23 (24% O1 serogroup isolates in streams and wells, whilst 64 (66.6% were non-O1/non-O139. Temperature and salinity correlated positively with the occurrence of the organisms. All isolates were susceptible to fluoroquinolones but high resistance rates to trimethoprim or sulfamethozaxole and tetracycline were observed.Vibrio cholerae is endemic in Bepanda with O1 and non-O1/non-O139 serogroups co-existing in the streams and wells hence the possibility of future outbreaks of cholera if sanitation and drinking water quality are not improved. Temperature and salinity are amongst the factors maintaining the endemicity of the organism.

  2. ToxR of Vibrio cholerae affects biofilm, rugosity and survival with Acanthamoeba castellanii

    Directory of Open Access Journals (Sweden)

    Valeru Soni P

    2012-01-01

    Full Text Available Abstract Background Vibrio cholerae causes the diarrheal disease cholera and utilizes different survival strategies in aquatic environments. V. cholerae can survive as free-living or in association with zooplankton and can build biofilm and rugose colonies. The bacterium expresses cholera toxin (CT and toxin-coregulated pilus (TCP as the main virulence factors. These factors are co-regulated by a transcriptional regulator ToxR, which modulates expression of outer membrane proteins (OmpU and (OmpT. The aims of this study were to disclose the role of ToxR in expression of OmpU and OmpT, biofilm and rugose colony formation as well as in association with the free-living amoeba Acanthamoeba castellanii at different temperatures. Results The toxR mutant V. cholerae produced OmpT, significant biofilm and rugose colonies compared to the wild type that produced OmpU, decreased biofilm and did not form rugoes colonies at 30°C. Interestingly, neither the wild type nor toxR mutant strain could form rugose colonies in association with the amoebae. However, during the association with the amoebae it was observed that A. castellanii enhanced survival of V. cholerae wild type compared to toxR mutant strain at 37°C. Conclusions ToxR does seem to play some regulatory role in the OmpT/OmpU expression shift, the changes in biofilm, rugosity and survival with A. castellanii, suggesting a new role for this regulatory protein in the environments.

  3. Genome engineering in Vibrio cholerae

    DEFF Research Database (Denmark)

    Val, Marie-Eve; Skovgaard, Ole; Ducos-Galand, Magaly;

    2012-01-01

    Although bacteria with multipartite genomes are prevalent, our knowledge of the mechanisms maintaining their genome is very limited, and much remains to be learned about the structural and functional interrelationships of multiple chromosomes. Owing to its bi-chromosomal genome architecture and its....... 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...

  4. [Cholera in a district of Peru].

    Science.gov (United States)

    Pérez Rodríguez, A E; Monté Boada, R; de la Vega Díaz, J E; Molina, R; García Gómez, V; Arca González, J M

    1996-01-01

    Taking to consideration the low report of cholera patients and with the main knowing the reality about the introduction of Vibrio cholerae (V. cholerae) in Peru, a sample of 101 cases with acute diarrheal disease (ADD) was taken at the Distrito Villa El Salvador. They were selected by a systematic randomized sampling defined for each health care unit in the District, according to the daily average occurrence of ADD cases attended a week before the beginning of the study. All of them took part in a epidemiological survey. A sample was taken by rectal swab in order to isolate V. cholerae. 53 positive cases were found (52.2% and a confidence interval from 42.29 to 62.5%) with significant differences (p < 0.01) between the frequency in adults (67.3%) and children (34.8%). V. cholerae was isolated only in 13 (61.9%) of the 21 cases who had contact with cholera patients, for a relative risk of 1.24 (0.83 < RR < 1.85). A high positivity was also found, 21 cases (72.4%) among those who had raw food. A significant difference (p < 0.01) was observed in connection with those who had cooked food. In the multivariate logistic regression analysis it was only found a significant relationship with age and with the ingestion of raw food as regards the occurrence of cholera. PMID:9805053

  5. The antimicrobial activity of ZnO nanoparticles against Vibrio cholerae: Variation in response depends on biotype.

    Science.gov (United States)

    Sarwar, Shamila; Chakraborti, Soumyananda; Bera, Supriyo; Sheikh, Irshad Ali; Hoque, Kazi Mirajul; Chakrabarti, Pinak

    2016-08-01

    The potency of zinc oxide nanoparticles (NPs), with a core size of ~7-10nm, to inhibit cholera disease was investigated by demonstrating the effect on two biotypes (classical and El Tor) of O1 serogroup of Vibrio cholerae-El Tor was more susceptible both in planktonic and in biofilm forms. Interaction with ZnO NP results in deformed cellular architecture. Increased fluidity and depolarization of membrane, and protein leakage further confirmed the damages inflicted on Vibrio by NP. NP was shown to produce reactive oxygen species (ROS) and induce DNA damage. These results suggest that the antibacterial mechanism of ZnO action is most likely due to generation of ROS and disruption of bacterial membrane. The antimicrobial efficacy of NP has been validated in animal model. The synergistic action of NP and antibiotic suggests an alternative for the treatment of cholera. PMID:26970029

  6. Production of Vibrio cholerae accessory cholera enterotoxin (Ace) in the yeast Pichia pastoris.

    OpenAIRE

    Trucksis, M; Conn, T L; Fasano, A; Kaper, J B

    1997-01-01

    Accessory cholera enterotoxin (Ace) is a recently identified toxin of Vibrio cholerae. Preliminary studies using crude toxin extracts in animal models indicate that Ace increases transcellular ion transport, which is proposed to contribute to diarrhea in cholera. The lack of purified toxin has hindered elucidation of the mechanism of action of Ace. In this study, ace was cloned and was expressed in and secreted by the methylotrophic yeast Pichia pastoris. Secreted toxin constituted 50% of the...

  7. Comparative genomic analysis reveals evidence of two novel Vibrio species closely related to V. cholerae

    Directory of Open Access Journals (Sweden)

    Brettin Thomas S

    2010-05-01

    Full Text Available Abstract Background In recent years genome sequencing has been used to characterize new bacterial species, a method of analysis available as a result of improved methodology and reduced cost. Included in a constantly expanding list of Vibrio species are several that have been reclassified as novel members of the Vibrionaceae. The description of two putative new Vibrio species, Vibrio sp. RC341 and Vibrio sp. RC586 for which we propose the names V. metecus and V. parilis, respectively, previously characterized as non-toxigenic environmental variants of V. cholerae is presented in this study. Results Based on results of whole-genome average nucleotide identity (ANI, average amino acid identity (AAI, rpoB similarity, MLSA, and phylogenetic analysis, the new species are concluded to be phylogenetically closely related to V. cholerae and V. mimicus. Vibrio sp. RC341 and Vibrio sp. RC586 demonstrate features characteristic of V. cholerae and V. mimicus, respectively, on differential and selective media, but their genomes show a 12 to 15% divergence (88 to 85% ANI and 92 to 91% AAI compared to the sequences of V. cholerae and V. mimicus genomes (ANI Vibrio sp. RC341 and Vibrio sp. RC586 share 2104 ORFs (59% and 2058 ORFs (56% with the published core genome of V. cholerae and 2956 (82% and 3048 ORFs (84% with V. mimicus MB-451, respectively. The novel species share 2926 ORFs with each other (81% Vibrio sp. RC341 and 81% Vibrio sp. RC586. Virulence-associated factors and genomic islands of V. cholerae and V. mimicus, including VSP-I and II, were found in these environmental Vibrio spp. Conclusions Results of this analysis demonstrate these two environmental vibrios, previously characterized as variant V. cholerae strains, are new species which have evolved from ancestral lineages of the V. cholerae and V. mimicus clade. The presence of conserved integration loci for genomic islands as well as evidence of horizontal gene transfer between these two new

  8. Comparison of DOT-ELISA and Standard-ELISA for Detection of the Vibrio cholerae Toxin in Culture Supernatants of Bacteria Isolated from Human and Environmental Samples.

    Science.gov (United States)

    Meza-Lucas, Antonio; Pérez-Villagómez, María-Fernanda; Martínez-López, José-Patricio; García-Rodea, Ricardo; Martínez-Castelán, María-Guadalupe; Escobar-Gutiérrez, Alejandro; de-la-Rosa-Arana, Jorge-Luis; Villanueva-Zamudio, Altagracia

    2016-09-01

    A comparison of DOT-ELISA and Standard-ELISA was made for detection of Vibrio cholerae toxin in culture supernatants of bacteria isolated from human and environmental samples. A total of 293 supernatants were tested in a double blind assay. A correlation of 100 % was obtained between both techniques. The cholera toxin was found in 20 Inaba and 3 Ogawa strains. Positive samples were from seafood (17 samples), potable water (1 sample) and sewage (5 samples). The DOT-ELISA was useful as the standard-ELISA to confirm the presence of cholera toxin in the environmental samples. PMID:27407304

  9. 进出口水产品中霍乱弧菌的检测与分析%Detection and analysis of Vibrio cholerae from import and export aquatic products

    Institute of Scientific and Technical Information of China (English)

    李正义; 贾俊涛; 梁成珠; 卢丽丽; 姜英辉; 祝素珍; 赵丽青; 房保海; 雷质文

    2011-01-01

    目的 监测并分析进出口水产品中的霍乱弧菌.方法 采用传统两步增菌,平板划线,分离可疑单菌落,VITEK 2 Compact全自动微生物鉴定仪、普通PCR、实时荧光定量PCR和血清凝集试验确认分离菌株.结果 从水产品样品分离的菌株全自动微生物鉴定仪鉴定结果相似度达97%,普通PCR和实时荧光定量PCR检测为霍乱弧菌,ctx毒素基因检测为阴性,血清凝集试验为非O1非O139群霍乱弧菌.结论 从水产品中检出霍乱弧菌非流行株,但仍需要加强主动监测,预防疫情的暴发.%Objective To monitor and analysis Vibrio cholerae from import and export aquatic products. Methods The suspicious single colonies by traditional two step enrichment and flat crossed separation was adopted. After isolation, the suspicious single colonies were identified by VITEK 2 Compact automated microbe identification analyzer, common PCR, real-time PCR and serological tests. Results The probability of the isolated strain was 97% by VITEK 2 Compact automated microbe identification analyzer. It was detected as Vibrio cholera by common PCR and real-time PCR and the primer and fluorescent probe were from industrial standard. The virulence gene ctx in the isolated strain was not : detected by common PCR. It was confirmed as non-O1 and non-O139 Vibrio cholera by serum agglutination test. Conclusion Although one non-epidemic strain of Vibrio chloerae was detected from the monitoring aquatic products, the monitoring work should be strengthened for epidemic prevention.

  10. DNA damage and reactive nitrogen species are barriers to Vibrio cholerae colonization of the infant mouse intestine.

    Directory of Open Access Journals (Sweden)

    Bryan W Davies

    2011-02-01

    Full Text Available Ingested Vibrio cholerae pass through the stomach and colonize the small intestines of its host. Here, we show that V. cholerae requires at least two types of DNA repair systems to efficiently compete for colonization of the infant mouse intestine. These results show that V. cholerae experiences increased DNA damage in the murine gastrointestinal tract. Agreeing with this, we show that passage through the murine gut increases the mutation frequency of V. cholerae compared to liquid culture passage. Our genetic analysis identifies known and novel defense enzymes required for detoxifying reactive nitrogen species (but not reactive oxygen species that are also required for V. cholerae to efficiently colonize the infant mouse intestine, pointing to reactive nitrogen species as the potential cause of DNA damage. We demonstrate that potential reactive nitrogen species deleterious for V. cholerae are not generated by host inducible nitric oxide synthase (iNOS activity and instead may be derived from acidified nitrite in the stomach. Agreeing with this hypothesis, we show that strains deficient in DNA repair or reactive nitrogen species defense that are defective in intestinal colonization have decreased growth or increased mutation frequency in acidified nitrite containing media. Moreover, we demonstrate that neutralizing stomach acid rescues the colonization defect of the DNA repair and reactive nitrogen species defense defective mutants suggesting a common defense pathway for these mutants.

  11. Nanostructured magnesium oxide biosensing platform for cholera detection

    Science.gov (United States)

    Patel, Manoj K.; Azahar Ali, Md.; Agrawal, Ved V.; Ansari, Z. A.; Ansari, S. G.; Malhotra, B. D.

    2013-04-01

    We report fabrication of highly crystalline nanostructured magnesium oxide (NanoMgO, size >30 nm) film electrophoretically deposited onto indium-tin-oxide (ITO) glass substrate for Vibrio cholerae detection. The single stranded deoxyribonucleic acid (ssDNA) probe, consisting of 23 bases (O1 gene sequence) immobilized onto NanoMgO/ITO electrode surface, has been characterized using electrochemical, Fourier Transform-Infra Red, and UltraViolet-visible spectroscopic techniques. The hybridization studies of ssDNA/NanoMgO/ITO bioelectrode with fragmented target DNA conducted using differential pulse voltammetry reveal sensitivity as 16.80 nA/ng/cm2, response time of 3 s, linearity as 100-500 ng/μL, and stability of about 120 days.

  12. Diversity of Clinical and Environmental Isolates of Vibrio cholerae in Natural Transformation and Contact-Dependent Bacterial Killing Indicative of Type VI Secretion System Activity.

    Science.gov (United States)

    Bernardy, Eryn E; Turnsek, Maryann A; Wilson, Sarah K; Tarr, Cheryl L; Hammer, Brian K

    2016-05-01

    The bacterial pathogen Vibrio cholerae can occupy both the human gut and aquatic reservoirs, where it may colonize chitinous surfaces that induce the expression of factors for three phenotypes: chitin utilization, DNA uptake by natural transformation, and contact-dependent bacterial killing via a type VI secretion system (T6SS). In this study, we surveyed a diverse set of 53 isolates from different geographic locales collected over the past century from human clinical and environmental specimens for each phenotype outlined above. The set included pandemic isolates of serogroup O1, as well as several serogroup O139 and non-O1/non-O139 strains. We found that while chitin utilization was common, only 22.6% of the isolates tested were proficient at chitin-induced natural transformation, suggesting that transformation is expendable. Constitutive contact-dependent killing of Escherichia coli prey, which is indicative of a functional T6SS, was rare among clinical isolates (only 4 of 29) but common among environmental isolates (22 of 24). These results bolster the pathoadaptive model in which tight regulation of T6SS-mediated bacterial killing is beneficial in a human host, whereas constitutive killing by environmental isolates may give a competitive advantage in natural settings. Future sequence analysis of this set of diverse isolates may identify previously unknown regulators and structural components for both natural transformation and T6SS. PMID:26944842

  13. Signaling beyond Punching Holes: Modulation of Cellular Responses by Vibrio cholerae Cytolysin

    Directory of Open Access Journals (Sweden)

    Barkha Khilwani

    2015-08-01

    Full Text Available Pore-forming toxins (PFTs are a distinct class of membrane-damaging cytolytic proteins that contribute significantly towards the virulence processes employed by various pathogenic bacteria. Vibrio cholerae cytolysin (VCC is a prominent member of the beta-barrel PFT (beta-PFT family. It is secreted by most of the pathogenic strains of the intestinal pathogen V. cholerae. Owing to its potent membrane-damaging cell-killing activity, VCC is believed to play critical roles in V. cholerae pathogenesis, particularly in those strains that lack the cholera toxin. Large numbers of studies have explored the mechanistic basis of the cell-killing activity of VCC. Consistent with the beta-PFT mode of action, VCC has been shown to act on the target cells by forming transmembrane oligomeric beta-barrel pores, thereby leading to permeabilization of the target cell membranes. Apart from the pore-formation-induced direct cell-killing action, VCC exhibits the potential to initiate a plethora of signal transduction pathways that may lead to apoptosis, or may act to enhance the cell survival/activation responses, depending on the type of target cells. In this review, we will present a concise view of our current understanding regarding the multiple aspects of these cellular responses, and their underlying signaling mechanisms, evoked by VCC.

  14. Temperature affects c-di-GMP signalling and biofilm formation in Vibrio cholerae.

    Science.gov (United States)

    Townsley, Loni; Yildiz, Fitnat H

    2015-11-01

    Biofilm formation is crucial to the environmental survival and transmission of Vibrio cholerae, the facultative human pathogen responsible for the disease cholera. During its infectious cycle, V. cholerae experiences fluctuations in temperature within the aquatic environment and during the transition between human host and aquatic reservoirs. In this study, we report that biofilm formation is induced at low temperatures through increased levels of the signalling molecule, cyclic diguanylate (c-di-GMP). Strains harbouring in frame deletions of all V. cholerae genes that are predicted to encode diguanylate cyclases (DGCs) or phosphodiesterases (PDEs) were screened for their involvement in low-temperature-induced biofilm formation and Vibrio polysaccharide gene expression. Of the 52 mutants tested, deletions of six DGCs and three PDEs were found to affect these phenotypes at low temperatures. Unlike wild type, a strain lacking all six DGCs did not exhibit a low-temperature-dependent increase in c-di-GMP, indicating that these DGCs are required for temperature modulation of c-di-GMP levels. We also show that temperature modulates c-di-GMP levels in a similar fashion in the Gram-negative pathogen Pseudomonas aeruginosa but not in the Gram-positive pathogen Listeria monocytogenes. This study uncovers the role of temperature in environmental regulation of biofilm formation and c-di-GMP signalling.

  15. Pandemic spread of cholera: genetic diversity and relationships within the seventh pandemic clone of Vibrio cholerae determined by amplified fragment length polymorphism.

    Science.gov (United States)

    Lan, Ruiting; Reeves, Peter R

    2002-01-01

    The seventh cholera pandemic started in 1961 and continues today. A collection of 45 seventh pandemic isolates of V. cholerae sampled over a 33-year period were analyzed by amplified fragment length polymorphism (AFLP) fingerprinting. All but four pairs and one set of three isolates were distinguished. AFLP revealed far more variation than ribotyping, which was until now the most useful method of revealing variation within the pandemic clone. Unfortunately, the ribotype variation observed is mainly due to recombination between the multiple copies of the rrn genes (R. Lan and P. R. Reeves, Microbiology 144:1213-1221, 1998), which makes changes susceptible to repeat occurrences and reversion. This AFLP study shows that particularly for the common ribotypes G and H, such events have indeed occurred. AFLP grouped most of the 45 isolates into two clusters. Cluster I consists mainly of strains from the 1960s and 1970s, while cluster II contains mainly strains from the 1980s and 1990s, revealing a temporal pattern of change in the clone. This is best seen in the relationships of the strains from Africa, which correlate with the epidemiology of epidemics on that continent. The data confirm independent introductions to Africa during the 1970s outbreak and reveal several other African introductions. In the 1991 cholera upsurge, isolates from the Southern and Eastern African epidemic focus are markedly different from those from the West African epidemic focus. An isolate from 1987 in Algeria was identical to the West epidemic isolates, suggesting that the strain was present in Africa at least 3 years before causing large outbreaks. These observations have major implications for our understanding of cholera epidemiology. PMID:11773113

  16. Evaluation of a bivalent (CVD 103-HgR/CVD 111) live oral cholera vaccine in adult volunteers from the United States and Peru.

    Science.gov (United States)

    Taylor, D N; Tacket, C O; Losonsky, G; Castro, O; Gutierrez, J; Meza, R; Nataro, J P; Kaper, J B; Wasserman, S S; Edelman, R; Levine, M M; Cryz, S J

    1997-09-01

    To provide optimum protection against classical and El Tor biotypes of Vibrio cholerae O1, a single-dose, oral cholera vaccine was developed by combining two live, attenuated vaccine strains, CVD 103-HgR (classical, Inaba) and CVD 111 (El Tor, Ogawa). The vaccines were formulated in a double-chamber sachet; one chamber contained lyophilized bacteria, and the other contained buffer. In the first study, 23 U.S. adult volunteers received CVD 103-HgR at 10(8) CFU plus CVD 111 at 10(8), 10(7), or 10(6) CFU, CVD 111 alone at 10(7) CFU, or placebo. In the second study, 275 Peruvian adults were randomized to receive CVD 103-HgR at 10(9) CFU plus CVD 111 at 10(9) or 10(8) CFU, CVD 111 alone at 10(9) CFU, CVD 103-HgR alone at 10(9) CFU, or placebo. Three of 15 U.S. volunteers who received CVD 111 at 10(7) or 10(8) CFU developed mild diarrhea, compared to none of 4 who received CVD 111 at 10(6) CFU and 1 of 4 who received placebo. Twelve (63%) of 19 vaccine recipients shed the El Tor vaccine strain. All but one volunteer developed significant Ogawa and Inaba vibriocidal antibody titers. Volunteers who received CVD 111 at 10(7) CFU had geometric mean Ogawa titers four to five times higher than those of volunteers who received the lower dose. In the second study, all dosage regimens were well tolerated in Peruvians. About 20% of volunteers who received CVD 111 at the high dose excreted the El Tor organism, compared to 7% in the low-dose group. CVD 111 was detected in the stools of two placebo recipients, neither of whom had symptoms or seroconverted. In all vaccine groups, 69 to 76% developed fourfold rises in Inaba vibriocidal antibodies. Among those who received the bivalent vaccine, 53 to 75% also developed significant rises in Ogawa vibriocidal antibodies. We conclude that it is feasible to produce a single-dose, oral bivalent vaccine that is safe and immunogenic against both biotypes (El Tor and classical) and both serotypes (Inaba and Ogawa) of cholera for populations in

  17. EFFECT OF AGGREGATION ON VIBRIO CHOLERA INACTIVATION

    Science.gov (United States)

    Extensive research has shown that microorganisms exhibit increased resistance due to clumping, aggregation, particle association or modification of antecedent growth conditions. uring the course of investigating a major waterborne V. Cholerae outbreak in Peru, U.S. EPA investigat...

  18. FOWL CHOLERA IN A BREEDER FLOCK

    OpenAIRE

    Z. Parveen, A. A. Nasir, K.Tasneem and A. Shah

    2003-01-01

    During January, 2003 Pasteurella multocida the causative agent of fowl cholera was isolated from a breeder flock in Lahore District. The age of the flock was 245 days. Increased mortality, swollen wattles and lameness were the clinical findings present in almost all the affected birds, while gross lesions were typical of fowl cholera. To prove the virulence of the organism, mice and six-week old cockerals were infected and P. multocida was reisolated.

  19. A genomic island integrated into recA of Vibrio cholerae contains a divergent recA and provides multi-pathway protection from DNA damage.

    Science.gov (United States)

    Rapa, Rita A; Islam, Atiqul; Monahan, Leigh G; Mutreja, Ankur; Thomson, Nicholas; Charles, Ian G; Stokes, Harold W; Labbate, Maurizio

    2015-04-01

    Lateral gene transfer (LGT) has been crucial in the evolution of the cholera pathogen, Vibrio cholerae. The two major virulence factors are present on two different mobile genetic elements, a bacteriophage containing the cholera toxin genes and a genomic island (GI) containing the intestinal adhesin genes. Non-toxigenic V. cholerae in the aquatic environment are a major source of novel DNA that allows the pathogen to morph via LGT. In this study, we report a novel GI from a non-toxigenic V. cholerae strain containing multiple genes involved in DNA repair including the recombination repair gene recA that is 23% divergent from the indigenous recA and genes involved in the translesion synthesis pathway. This is the first report of a GI containing the critical gene recA and the first report of a GI that targets insertion into a specific site within recA. We show that possession of the island in Escherichia coli is protective against DNA damage induced by UV-irradiation and DNA targeting antibiotics. This study highlights the importance of genetic elements such as GIs in the evolution of V. cholerae and emphasizes the importance of environmental strains as a source of novel DNA that can influence the pathogenicity of toxigenic strains.

  20. Etiological Characteristics and Traceability of Vibrio cholerae in Hunan Province in 2010%2010年湖南省霍乱弧菌病原学特征及溯源分析

    Institute of Scientific and Technical Information of China (English)

    覃迪; 湛志飞; 夏昕; 胡世雄; 邓志红; 刘运芝; 黄一伟; 龙智钢; 张红

    2012-01-01

    目的 分析2010年湖南省霍乱弧菌分离株的病原学特征,比较霍乱疫情分离株与常规监测分离株之间的克隆相关性,追溯传染源. 方法 对疫情与监测分离到的42株霍乱弧菌进行常规生物分型和PCR检测毒力基因,对23株代表株进行药敏试验,对18株代表株通过脉冲场凝胶电泳( PFGE)获得电泳图谱,利用BioNumerics软件对图谱进行聚类分析,探讨菌株间的相关性. 结果 2010年从湖南省霍乱疫情中分离10株霍乱弧菌均为O139群,ctxA阳性率100%.常规监测分离霍乱弧菌32株,其中O1群15株,全部为ctxA阴性株;O139群17株,ctxA阳性率94.11%.23株霍乱弧菌耐药结果显示强力霉素、复方新诺明的耐药率分别为47.83%、56.52%,发现1株对诺氟沙星、环丙沙星耐药.PFGE方法显示有5种脉冲场凝胶电泳图谱,相似率在82% ~ 100%之间,甲鱼中分离的O139群霍乱弧菌与霍乱疫情分离菌株之间高度同源. 结论 湖南省霍乱弧菌存在紧密相关的流行克隆群;被O139群霍乱弧菌污染的甲鱼很可能是湖南省霍乱疫情发生的主要传染来源,海冰产品的监测是霍乱防控的重点;要密切关注对诺氟沙星、环丙沙星的耐药变化.%Objective To analyze the pathogenic characteristics of Vibrio cholerae strains isolated in Hunan province in 2010, to compare cholera epidemic strains with routine monitoring strains in homology, and to trace the source of infection. Methods Forty-two strains isolated from epidemic and routine monitoring were tested for conventional biotyping and virulence gene by PCR. Twenty-three representative strains were tested for drug susceptibility. Electrophoresis maps were obtained from 18 representative strains by pulsed-field gel electrophoresis (PFGE) and analyzed for clusters and correlations by BioNumerics software. Results All of the 10 strains isolated from Vibrio cholerae outbreaks in Hunan province belonged to 0139 group. The

  1. BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING

    Directory of Open Access Journals (Sweden)

    Milagro FERNÁNDEZ-DELGADO

    2016-01-01

    Full Text Available Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM. Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS, while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing.

  2. BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING

    Science.gov (United States)

    FERNÁNDEZ-DELGADO, Milagro; ROJAS, Héctor; DUQUE, Zoilabet; SUÁREZ, Paula; CONTRERAS, Monica; GARCÍA-AMADO, M. Alexandra; ALCIATURI, Carlos

    2016-01-01

    Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM). Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS), while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing. PMID:27253749

  3. BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING.

    Science.gov (United States)

    Fernández-Delgado, Milagro; Rojas, Héctor; Duque, Zoilabet; Suárez, Paula; Contreras, Monica; García-Amado, M Alexandra; Alciaturi, Carlos

    2016-01-01

    Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM). Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS), while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing. PMID:27253749

  4. Sensitive and rapid detection of Vibrio cholerae O139 by loop-mediated isothermal amplification%应用环介导等温扩增法快速检测O139群霍乱弧菌

    Institute of Scientific and Technical Information of China (English)

    朱水荣; 陈寅; 王志刚; 张政; 朱敏; 卢亦愚

    2009-01-01

    Objective To develop a loop-mediated isothermal amplification method for rapid diag-nosing Vibrio cholerae O139 in inspection department and small-scale laboratory. Methods Four primers (2 inner primer and 2 outer primer) for the LAMP test were designed by targeting the wbfR gene of Vibrio chol-erae O139, and the reaction condition and reaction system of LAMP were optimized. Thirty Vibrio cholerae O139, 13 Vibrio cholerae reference strains, 10 O1 biotype Vibrio cholera* and 32 other enterobacterias were analyzed and the LAMP results were determined by visual inspection or electrophoretie analysis . Results All of the Vibrio cholerae O139's amplification products were observed as green by visual inspection and had a ladder-like pattern on the gel, but O1 biotype Vibrio cholera* and other enterobacteria's products were dis-played as orange by visual examination and had no ladder-like pattern on the gel. In addition, the reaction time of the LAMP method was only 1.5 h and the detection limit of this assay was 63 CFU/reaction. Con-clnsion LAMP assay targeting the wbfR gene of Vibrio cholera* O139 is rapid, specific, and sensitive for the detection of Vibrio cholerae O139. This method not only reduced the dependence of complicated equip-ment but also had a potential for wider use in inspection department, small-scale laboratory, emergency mo-tor vehicles and field survey.%目的 建立一种适合基层检验部门及小型实验室使用的快速检测O139群霍乱弧菌的方法.方法 应用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP),针对O139霍乱弧菌wbfR基因设计4条引物(2条内引物、2条外引物);优化LAMP反应条件和反应体系,对反应体系中的引物、dNTP、Mg2+/Mn2+离子及Calcium等浓度进行优化;并对13株种系背景明确的霍乱弧菌不同实验对照株、30株O139群霍乱弧菌地方分离株、10株O1群霍乱弧菌地方分离株、32株其他肠道菌进行检测,验证该方法的特异

  5. Enhancement of enterotoxin production by carbon dioxide in Vibrio cholerae.

    OpenAIRE

    Shimamura, T; Watanabe, S; Sasaki, S.

    1985-01-01

    We found that Vibrio cholerae 569B produced much more cholera enterotoxin in the presence of added carbon dioxide than in its absence. An atmosphere of 10% carbon dioxide was optimal for maximal enterotoxin production.

  6. 萍乡地区O139群霍乱弧菌TcpA基因序列分析%Gene Sequencing Analysis on Vibrio Cholera O139 Strains TcpA of Pingxiang Area

    Institute of Scientific and Technical Information of China (English)

    王长奇; 姚彬泉; 贺丁山; 陈燕萍; 罗娜; 周文峰

    2003-01-01

    目的探讨O139群霍乱JX株的TcpA基因序列,分析该株基因的特点.方法采用PCR扩增技术,检测含O139群霍乱标本,对TcpA基因阳性者进行核苷酸序列测定,分析基因变异情况.结果发现阳性标本中,对其中1株进行了TcpA基因序列测定,该序列与TcpAET(O139株)同源性为100%,与EVC序列同源性为96%.结论本株与O1群(EVC)遗传关系最近,对萍乡地区霍乱防治具有重要意义.

  7. Repair of ultraviolet-light-induced DNA damage in Vibrio cholerae

    International Nuclear Information System (INIS)

    Repair of ultraviolet-light-induced DNA damage in a highly pathogenic Gram-negative bacterium, Vibrio cholerae, has been examined. All three strains of V. cholerae belonging to two serotypes, Inaba and Ogawa, are very sensitive to ultraviolet irradiation, having inactivation cross-sections ranging from 0.18 to 0.24 m2/J. Although these cells are proficient in repairing the DNA damage by a photoreactivation mechanism, they do not possess efficient dark repair systems. The mild toxinogenic strain 154 of classical Vibrios presumably lacks any excision repair mechanism and studies of irradiated cell DNA indicate that the ultraviolet-induced pyrimidine dimers may not be excised. Ultraviolet-irradiated cells after saturation of dark repair can be further photoreactivated. (Auth.)

  8. Highly Potent, Chemically Stable Quorum Sensing Agonists for Vibrio Cholerae

    OpenAIRE

    Perez, Lark J; Karagounis, Theodora K.; Hurley, Amanda; Bassler, Bonnie L.; Semmelhack, Martin F.

    2013-01-01

    In the Vibrio cholerae pathogen, initiation of bacterial quorum sensing pathways serves to suppress virulence. We describe herein a potent and chemically stable small molecule agonist of V. cholerae quorum sensing, which was identified through rational drug design based on the native quorum sensing signal. This novel agonist may serve as a useful lead compound for the control of virulence in V. cholerae.

  9. 21 CFR 866.3930 - Vibrio cholerae serological reagents.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Vibrio cholerae serological reagents. 866.3930... cholerae serological reagents. (a) Identification. Vibrio cholerae serological reagents are devices that are used in the agglutination (an antigen-antibody clumping reaction) test to identify Vibrio...

  10. Bile Salts Modulate the Mucin-Activated Type VI Secretion System of Pandemic Vibrio cholerae.

    Science.gov (United States)

    Bachmann, Verena; Kostiuk, Benjamin; Unterweger, Daniel; Diaz-Satizabal, Laura; Ogg, Stephen; Pukatzki, Stefan

    2015-01-01

    The causative agent of cholera, Vibrio cholerae, regulates its diverse virulence factors to thrive in the human small intestine and environmental reservoirs. Among this pathogen's arsenal of virulence factors is the tightly regulated type VI secretion system (T6SS). This system acts as an inverted bacteriophage to inject toxins into competing bacteria and eukaryotic phagocytes. V. cholerae strains responsible for the current 7th pandemic activate their T6SS within the host. We established that T6SS-mediated competition occurs upon T6SS activation in the infant mouse, and that this system is functional under anaerobic conditions. When investigating the intestinal host factors mucins (a glycoprotein component of mucus) and bile for potential regulatory roles in controlling the T6SS, we discovered that once mucins activate the T6SS, bile acids can further modulate T6SS activity. Microbiota modify bile acids to inhibit T6SS-mediated killing of commensal bacteria. This interplay is a novel interaction between commensal bacteria, host factors, and the V. cholerae T6SS, showing an active host role in infection.

  11. Bile Salts Modulate the Mucin-Activated Type VI Secretion System of Pandemic Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    Verena Bachmann

    Full Text Available The causative agent of cholera, Vibrio cholerae, regulates its diverse virulence factors to thrive in the human small intestine and environmental reservoirs. Among this pathogen's arsenal of virulence factors is the tightly regulated type VI secretion system (T6SS. This system acts as an inverted bacteriophage to inject toxins into competing bacteria and eukaryotic phagocytes. V. cholerae strains responsible for the current 7th pandemic activate their T6SS within the host. We established that T6SS-mediated competition occurs upon T6SS activation in the infant mouse, and that this system is functional under anaerobic conditions. When investigating the intestinal host factors mucins (a glycoprotein component of mucus and bile for potential regulatory roles in controlling the T6SS, we discovered that once mucins activate the T6SS, bile acids can further modulate T6SS activity. Microbiota modify bile acids to inhibit T6SS-mediated killing of commensal bacteria. This interplay is a novel interaction between commensal bacteria, host factors, and the V. cholerae T6SS, showing an active host role in infection.

  12. Quorum regulatory small RNAs repress type VI secretion in Vibrio cholerae.

    Science.gov (United States)

    Shao, Yi; Bassler, Bonnie L

    2014-06-01

    Type VI secretion is critical for Vibrio cholerae to successfully combat phagocytic eukaryotes and to survive in the presence of competing bacterial species. V. cholerae type VI secretion system genes are encoded in one large and two small clusters. In V. cholerae, type VI secretion is controlled by quorum sensing, the cell-cell communication process that enables bacteria to orchestrate group behaviours. The quorum-sensing response regulator LuxO represses type VI secretion genes at low cell density and the quorum-sensing regulator HapR activates type VI secretion genes at high cell density. We demonstrate that the quorum regulatory small RNAs (Qrr sRNAs) that function between LuxO and HapR in the quorum-sensing cascade are required for these regulatory effects. The Qrr sRNAs control type VI secretion via two mechanisms: they repress expression of the large type VI secretion system cluster through base pairing and they repress HapR, the activator of the two small type VI secretion clusters. This regulatory arrangement ensures that the large cluster encoding many components of the secretory machine is expressed prior to the two small clusters that encode the secreted effectors. Qrr sRNA-dependent regulation of the type VI secretion system is conserved in pandemic and non-pandemic V. cholerae strains.

  13. A mutation in the dam gene of Vibrio cholerae: 2-aminopurine sensitivity with intact GATC methylase activity

    International Nuclear Information System (INIS)

    Vibrio cholerae mutants sensitive to 2-aminopurine (2AP) but with DNA adenine methylase activity similar to parental cells have been isolated. The mutant strains were sensitive to ultraviolet light (UV), methyl methanesulfonate (MMS) and 9-aminoacridine. The spontaneous mutation frequency of the mutants were not significantly affected. Attempts to isolate dam V. cholerae cells by screening 2AP sensitive cells have not been successful. All the mutant phenotypes could be suppressed by introducing the plasmid pRB103 carrying the dam gene of Escherichia coli into the mutant cells

  14. Development of a live, oral, attenuated vaccine against El Tor cholera.

    Science.gov (United States)

    Taylor, D N; Killeen, K P; Hack, D C; Kenner, J R; Coster, T S; Beattie, D T; Ezzell, J; Hyman, T; Trofa, A; Sjogren, M H

    1994-12-01

    Vibrio cholerae El Tor strains from Peru, Bangladesh, and Bahrain were attenuated by deletion of a genetic element that encodes virulence factors and RS1. The B subunit of ctx (ctxB) was reintroduced into the recA gene of the deletion mutants, rendering them unable to recombine with exogenous genetic elements and generating Peru-3, Bang-3, and Bah-3. Fifteen volunteers received one dose of various vaccine strains at 4 x 10(6) to 1 x 10(8) cfu. All strains colonized the gut. A > or = 4-fold rise in vibriocidal titer was observed in 14 volunteers, with titers of > or = 1600 in 13. Peru-3 was the least reactogenic, but 2 of 6 volunteers had loose stools. Peru-14, a filamentous motility-deficient mutant of Peru-3, was well tolerated and colonized 18 of 21 volunteers at doses of 2 x 10(6) to 1 x 10(9) cfu. Also, when 8 Peru-3 or Peru-5 vaccinees, 5 Peru-14 vaccinees, and 8 controls were challenged with 2 x 10(6) cfu V. cholerae El Tor Inaba (N16961), 11 vaccinees were protected compared with no controls. Peru-14 shows promise as a safe, effective, single-dose oral vaccine against El Tor cholera.

  15. Cloning of Vibrio cholerae outer membrane protein W in Pichia pastoris.

    Directory of Open Access Journals (Sweden)

    Javad Alizadeh

    2013-09-01

    Full Text Available The outer membrane protein W (ompW of Vibrio cholerae is involved in stimulating the immune response via induction of protective immunity. It also plays an important role in bacterial pathogenesis by increasing the adaptability of pathogenic strains. In this study we aimed to clone V. cholerae ompW gene in the strain X-33 of Pichia pastoris.A gene encoding ompW was cloned into the Ppicza vector downstream of alcohol oxidase promoter. Then recombinant vector was transformed into the genome of the strain X-33 of P. pastoris. After growth of zeocin-resistant transformants, clones were selected and subsequently confirmed for cloning by PCR enzymatic digestion and sequencing.PCR, enzymatic digestion and sequencing showed that the ompW gene was correctly cloned into P. pastoris genome.Results of our study showed that the methylotrophic yeast P. pastoris can be considered as an appropriate host instead of mammalian and prokaryotic systems for cloning of ompW. As far as data show, this is the first time that ompW of V. cholera is cloned into the methylotrophic P. pastoris.

  16. Computer based screening for novel inhibitors against Vibrio cholerae using NCI diversity set-II: an alternative approach by targeting transcriptional activator ToxT.

    Science.gov (United States)

    Mondal, Shakhinur Islam; Khadka, Bijendra; Akter, Arzuba; Roy, Pradip Kumar; Sultana, Razia

    2014-06-01

    Cholera is a severe diarrheal disease caused by Vibrio cholerae and remains as a major health risk in developing countries. The emergence and spread of multi-drug resistant V. cholerae strains during the past two decades is now a major problem in the treatment of cholera and have created the urgent need for the development of novel therapeutic agents. Targeting transcriptional factor is now a novel approach to tackle the development of multi-drug resistant strain. In the recent year virtual high throughput screening has emerged as a widely accepted powerful technology in the identification of novel and diverse lead. This study provides new insight to the search for new potent and selective inhibitors that still remains necessary to avoid the risk of possible resistance and reduce toxicity and side effects of currently available cholera drugs. The publications of high resolution X-ray structure of V. cholerae ToxT has open the way to the structure based virtual screening to identify new small molecular inhibitors which still remain necessary to avoid the risk of possible resistance and reduce toxicity and side effects of currently available cholera drugs. In this study we have performed structure based virtual screening approach using NCI diversity set-II to look for novel inhibitor of ToxT and proposed eight candidate compounds with high scoring function. Thus from complex scoring and binding ability it is elucidated that these compounds could be the promising inhibitors or could be developed as novel lead compounds for drug design against cholera.

  17. Vibrio cholerae as a predator: lessons from evolutionary principles

    Directory of Open Access Journals (Sweden)

    Stefan ePukatzki

    2013-12-01

    Full Text Available Diarrheal diseases are the second-most common cause of death among children under the age of five worldwide. Cholera alone, caused by the marine bacterium Vibrio cholerae, is responsible for several million cases and over 120,000 deaths annually. When contaminated water is ingested, V. cholerae passes through the gastric acid barrier, penetrates the mucin layer of the small intestine, and adheres to the underlying epithelial lining. V. cholerae multiplies rapidly, secretes cholera toxin, and exits the human host in vast numbers during diarrheal purges. How V. cholerae rapidly reaches such high numbers during each purge is not clearly understood. We propose that V. cholerae employs its bactericidal type VI secretion system to engage in intraspecies and intraguild predation for nutrient acquisition to support rapid growth and multiplication.

  18. Population-Level Effect of Cholera Vaccine on Displaced Populations, South Sudan, 2014.

    Science.gov (United States)

    Azman, Andrew S; Rumunu, John; Abubakar, Abdinasir; West, Haley; Ciglenecki, Iza; Helderman, Trina; Wamala, Joseph Francis; Vázquez, Olimpia de la Rosa; Perea, William; Sack, David A; Legros, Dominique; Martin, Stephen; Lessler, Justin; Luquero, Francisco J

    2016-06-01

    Following mass population displacements in South Sudan, preventive cholera vaccination campaigns were conducted in displaced persons camps before a 2014 cholera outbreak. We compare cholera transmission in vaccinated and unvaccinated areas and show vaccination likely halted transmission within vaccinated areas, illustrating the potential for oral cholera vaccine to stop cholera transmission in vulnerable populations.

  19. Extinction of Vibrio cholerae in acidic substrata: contaminated cabbage and lettuce treated with lime juice.

    Science.gov (United States)

    Mata, L; Vargas, C; Saborío, D; Vives, M

    1994-12-01

    Lime juice killed millions of Vibrio cholerae O1, El Tor, Inaba, present on cabbage and lettuce contaminated in the laboratory. The lethal effect was evident within 5 min of exposure to lime juice. No vibrios could be recovered at dilution 1:10 using alkaline peptone water (APW) and thiosulfate-citrate-bile salts-saccharose agar (TCBS). More than 99.9% of the initial inoculum was effectively destroyed. The number of vibrios killed by lime juice was 2 to 6 logarithms greater than the maximum infecting dose, and 4 to 8 logs greater than the minimum infecting dose for cholera El Tor. The time interval needed for killing was smaller than the usual waiting time for serving food in homes and restaurants. The addition of lime juice to non-acidic foods, beverages and water, is strongly recommended to prevent infection with cholera vibrios and other acid-sensitive microorganisms. This measure is particularly important for rural and slum populations in the tropics and subtropics.

  20. Sequence analysis of toxin-coregulated pilus gene of Vibrio cholerae strains isolated from China%我国霍乱弧菌毒力协同调节菌毛基因序列分析

    Institute of Scientific and Technical Information of China (English)

    芮勇宇; 阚飙; 刘朋; 高守一; 刘延清; 祁国明

    2006-01-01

    目的对我国霍乱弧菌(VC)毒力协同调节菌毛(tcpA)基因进行序列分析.方法 38株VC的tcpA基因经聚合酶链反应扩增、测序和序列分析.结果 13株O139群VC产毒株和15株EVC产毒株(均为流行株)的tcpA类型与EVC标准株N16961为同一类型,共有4个位点碱基发生变异,分为5个亚型.EVC流行株中2 株非产毒株的tcpA类型,1株同N16961,1株为新类型.6株EVC非流行株均为非产毒株,分为2种新类型.2株非O1非O139群VC均为非产毒株,为2种新类型.4种新类型与国外4种类型基因和氨基酸序列比较,同源性为63.8%~84.6%和60.6%~91.7%,趋异性为17.8%~44.6%和8.8%~45.8%,tcpA基因所在的VPI毒力岛的结构是完整的,但部分基因存在一定变异.结论 O139群VC产毒株可能起源于EVC产毒株,然后再进一步分化;在非产毒VC中发现4种新类型tcpA基因.

  1. The case for reactive mass oral cholera vaccinations.

    Directory of Open Access Journals (Sweden)

    Rita Reyburn

    Full Text Available INTRODUCTION: The outbreak of cholera in Zimbabwe intensified interest in the control and prevention of cholera. While there is agreement that safe water, sanitation, and personal hygiene are ideal for the long term control of cholera, there is controversy about the role of newer approaches such as oral cholera vaccines (OCVs. In October 2009 the Strategic Advisory Group of Experts advised the World Health Organization to consider reactive vaccination campaigns in response to large cholera outbreaks. To evaluate the potential benefit of this pivotal change in WHO policy, we used existing data from cholera outbreaks to simulate the number of cholera cases preventable by reactive mass vaccination. METHODS: Datasets of cholera outbreaks from three sites with varying cholera endemicity--Zimbabwe, Kolkata (India, and Zanzibar (Tanzania--were analysed to estimate the number of cholera cases preventable under differing response times, vaccine coverage, and vaccine doses. FINDINGS: The large cholera outbreak in Zimbabwe started in mid August 2008 and by July 2009, 98,591 cholera cases had been reported with 4,288 deaths attributed to cholera. If a rapid response had taken place and half of the population had been vaccinated once the first 400 cases had occurred, as many as 34,900 (40% cholera cases and 1,695 deaths (40% could have been prevented. In the sites with endemic cholera, Kolkata and Zanzibar, a significant number of cases could have been prevented but the impact would have been less dramatic. A brisk response is required for outbreaks with the majority of cases occurring during the early weeks. Even a delayed response can save a substantial number of cases and deaths in long, drawn-out outbreaks. If circumstances prevent a rapid response there are good reasons to roll out cholera mass vaccination campaigns well into the outbreak. Once a substantial proportion of a population is vaccinated, outbreaks in subsequent years may be reduced if not

  2. Relationship between Distinct African Cholera Epidemics Revealed via MLVA Haplotyping of 337 Vibrio cholerae Isolates.

    Directory of Open Access Journals (Sweden)

    Sandra Moore

    Full Text Available Since cholera appeared in Africa during the 1970s, cases have been reported on the continent every year. In Sub-Saharan Africa, cholera outbreaks primarily cluster at certain hotspots including the African Great Lakes Region and West Africa.In this study, we applied MLVA (Multi-Locus Variable Number Tandem Repeat Analysis typing of 337 Vibrio cholerae isolates from recent cholera epidemics in the Democratic Republic of the Congo (DRC, Zambia, Guinea and Togo. We aimed to assess the relationship between outbreaks. Applying this method, we identified 89 unique MLVA haplotypes across our isolate collection. MLVA typing revealed the short-term divergence and microevolution of these Vibrio cholerae populations to provide insight into the dynamics of cholera outbreaks in each country. Our analyses also revealed strong geographical clustering. Isolates from the African Great Lakes Region (DRC and Zambia formed a closely related group, while West African isolates (Togo and Guinea constituted a separate cluster. At a country-level scale our analyses revealed several distinct MLVA groups, most notably DRC 2011/2012, DRC 2009, Zambia 2012 and Guinea 2012. We also found that certain MLVA types collected in the DRC persisted in the country for several years, occasionally giving rise to expansive epidemics. Finally, we found that the six environmental isolates in our panel were unrelated to the epidemic isolates.To effectively combat the disease, it is critical to understand the mechanisms of cholera emergence and diffusion in a region-specific manner. Overall, these findings demonstrate the relationship between distinct epidemics in West Africa and the African Great Lakes Region. This study also highlights the importance of monitoring and analyzing Vibrio cholerae isolates.

  3. Pentavalent outer membrane vesicles of Vibrio cholerae induce adaptive immune response and protective efficacy in both adult and passive suckling mice models.

    Science.gov (United States)

    Sinha, Ritam; Koley, Hemanta; Nag, Dhrubajyoti; Mitra, Soma; Mukhopadhyay, Asish K; Chattopadhyay, Brajadulal

    2015-03-01

    Recently, we demonstrated oral immunizations with single serotype outer membrane vesicles of Vibrio cholerae induced serogroup specific protective immunity in the RITARD model. In our present study, we advanced our research by formulating multi-serotype outer membrane vesicles, mixing the OMVs of five virulent V. cholerae strains. Four doses of oral immunization with cholera pentavalent outer membrane vesicles (CPMVs) induced V. cholerae specific B and T cell responses. CPMVs-immunized mice generated long lasting serum IgG, IgA, IgM as well as mucosal sIgA and also elicited a higher percentage of CD4+ T cell distribution in spleen. Our study revealed that in vitro CPMVs-activated dendritic cells were secreting T cell polarizing cytokines, IL-12p40, IL-4, IL-6 and IL-1β. Moreover, purified splenic CD4+ T cells of immunized mice also secreted IL-4, IL-13 and IL-17 cytokines, indicating the initiation of Th2 and Th17 cell mediated immune responses. CPMVs immunized adult female mice and their offspring were significantly protected from heterologous challenge with wild type V. cholerae. CPMVs could be exploited for the development of a novel non-living vaccine against circulating cholera in near future.

  4. EFFECT OF AGGREGATION ON VIBRIO CHOLERAE INACTIVATION

    Science.gov (United States)

    Extensive research has shown that microorganisms exhibit increased resistance due to clumping, aggregation, particle association, or modification of antecedent growth conditions. During the course of investigating a major water-borne Vibrio cholerae outbreak in Peru, U.S. EPA inv...

  5. Maladi Kolera 1 PSA (:30) (Cholera 1)

    Centers for Disease Control (CDC) Podcasts

    2010-02-18

    This is an important public health announcement about cholera symptoms and ways you can prevent the spread of disease. Language: Haitian Creole.  Created: 2/18/2010 by Centers for Disease Control and Prevention (CDC).   Date Released: 2/18/2010.

  6. Maladi Kolera 2 PSA (:30) (Cholera 2)

    Centers for Disease Control (CDC) Podcasts

    2010-02-18

    This is an important public health announcement about cholera prevention and food preparation tips you can use to prevent the spread of disease. Language: Haitian Creole.  Created: 2/18/2010 by Centers for Disease Control and Prevention (CDC).   Date Released: 2/18/2010.

  7. Surface-attachment sequence in Vibrio Cholerae

    Science.gov (United States)

    Utada, Andrew; Gibiansky, Maxsim; Wong, Gerard

    2013-03-01

    Vibrio cholerae is a gram-negative bacterium that causes the human disease cholera. It is found natively in brackish costal waters in temperate climates, where it attaches to the surfaces of a variety of different aquatic life. V. cholerae has a single polar flagellum making it highly motile, as well as a number of different pili types, enabling it to attach to both biotic and abiotic surfaces. Using in-house built tracking software we track all surface-attaching bacteria from high-speed movies to examine the early-time attachment profile of v. cholerae onto a smooth glass surface. Similar to previous work, we observe right-handed circular swimming trajectories near surfaces; however, in addition we see a host of distinct motility mechanisms that enable rapid exploration of the surface before forming a more permanent attachment. Using isogenic mutants we show that the motility mechanisms observed are due to a complex combination of hydrodynamics and pili-surface interactions. Lauga, E., DiLuzio, W. R., Whitesides, G. M., Stone, H. A. Biophys. J. 90, 400 (2006).

  8. Preventing Maritime Transfer of Toxigenic Vibrio cholerae

    Science.gov (United States)

    Slaten, Douglas D.; Marano, Nina; Tappero, Jordan W.; Wellman, Michael; Albert, Ryan J.; Hill, Vincent R.; Espey, David; Handzel, Thomas; Henry, Ariel; Tauxe, Robert V.

    2012-01-01

    Organisms, including Vibrio cholerae, can be transferred between harbors in the ballast water of ships. Zones in the Caribbean region where distance from shore and water depth meet International Maritime Organization guidelines for ballast water exchange are extremely limited. Use of ballast water treatment systems could mitigate the risk for organism transfer. PMID:23017338

  9. Ecología de Vibrio cholerae en relación al Fitoplancton y variables fisicoquímicas en ríos de Tucumán (Argentina Ecology of Vibrio cholerae in relation to phytoplankton and physico-chemical variables in rivers of Tucumán (Argentina

    Directory of Open Access Journals (Sweden)

    V. Mirande

    Full Text Available Vibrio cholerae muestra gran diversidad serológica en base a su antígeno somático O, conociéndose al menos 200 serogrupos. De éstos, solamente O1 y O139 son causantes de epidemias o pandemias. En Latinoamérica el serogrupo O1 reapareció en 1991, tras cien años de no presentar brotes en el continente. Esta bacteria sobrevive y se multiplica asociada al plancton, independientemente de la aparición de infecciones humanas. Desde la década del noventa, en Tucumán, se detectaron casos esporádicos de diarrea por Vibrio cholerae no-O1. El objetivo del presente trabajo fue estudiar la posible relación entre la presencia de especímenes de fitoplancton, variables fisicoquímicas y aislamientos de Vibrio cholerae en ríos de Tucumán. Se realizaron 18 campañas en los ríos Lules y Salí entre 2003-2005. Se estudiaron las variables fisicoquímicas del agua (pH, temperatura, conductividad y oxígeno disuelto, el fitoplancton (riqueza y frecuencia relativa y las cepas aisladas de V. cholerae. Los resultados evidenciaron diferencias en la calidad del agua, observándose períodos de anoxia en el río Salí. Las diatomeas sobresalieron en la mayoría de los meses y generalmente estuvieron en porcentajes superiores al 85 %. Sólo se aisló Vibrio cholerae no-O1, no-O139, detectándose más frecuentemente en los meses cálidos, con pH alcalino, aún con baja concentración de oxígeno.Vibrio cholerae shows a great serologic diversity in relation to his O somatic antigen and we know at least 200 serogroups. About these, only O1 and O139 are responsible of epidemics and pandemics. The serogroup O1 reemerged in Latin America in 1991 after being absent from the continent for nearly a century. This bacterium survives and grows up associated to plankton, independently of appearance of human infections. From 90 th decade, there were sporadic cases of diarrhea because of Vibrio cholerae O1 in Tucumán. The aims of this paper were to study the possible

  10. Pasteurella multocida from outbreaks of avian cholera in wild and captive birds in Denmark

    DEFF Research Database (Denmark)

    Pedersen, Karl; Dietz, Hans-Henrik; Jørgensen, J.C.;

    2003-01-01

    An outbreak of avian cholera was observed among wild birds in a few localities in Denmark in 2001. The highest mortalities were among breeding ciders (Somateria mollissima) and gulls (Larus spp.). Pulsed-field gel electrophoresis (PFGE) was conducted using ApaI and SmaI as restriction enzymes...... and restriction enzyme analysis (REA) using HpaII. The Pasteurella multocida subsp. multocida strain isolated from birds in this outbreak was indistinguishable from a strain that caused outbreaks in 1996 and 2003. Most isolates from domestic poultry had other PFGE patterns but some were indistinguishable from...

  11. Effects of Small Molecule Calcium-Activated Chloride Channel Inhibitors on Structure and Function of Accessory Cholera Enterotoxin (Ace of Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    Tanaya Chatterjee

    Full Text Available Cholera pathogenesis occurs due to synergistic pro-secretory effects of several toxins, such as cholera toxin (CTX and Accessory cholera enterotoxin (Ace secreted by Vibrio cholerae strains. Ace activates chloride channels stimulating chloride/bicarbonate transport that augments fluid secretion resulting in diarrhea. These channels have been targeted for drug development. However, lesser attention has been paid to the interaction of chloride channel modulators with bacterial toxins. Here we report the modulation of the structure/function of recombinant Ace by small molecule calcium-activated chloride channel (CaCC inhibitors, namely CaCCinh-A01, digallic acid (DGA and tannic acid. Biophysical studies indicate that the unfolding (induced by urea free energy increases upon binding CaCCinh-A01 and DGA, compared to native Ace, whereas binding of tannic acid destabilizes the protein. Far-UV CD experiments revealed that the α-helical content of Ace-CaCCinh-A01 and Ace-DGA complexes increased relative to Ace. In contrast, binding to tannic acid had the opposite effect, indicating the loss of protein secondary structure. The modulation of Ace structure induced by CaCC inhibitors was also analyzed using docking and molecular dynamics (MD simulation. Functional studies, performed using mouse ileal loops and Ussing chamber experiments, corroborate biophysical data, all pointing to the fact that tannic acid destabilizes Ace, inhibiting its function, whereas DGA stabilizes the toxin with enhanced fluid accumulation in mouse ileal loop. The efficacy of tannic acid in mouse model suggests that the targeted modulation of Ace structure may be of therapeutic benefit for gastrointestinal disorders.

  12. Effects of Small Molecule Calcium-Activated Chloride Channel Inhibitors on Structure and Function of Accessory Cholera Enterotoxin (Ace) of Vibrio cholerae

    Science.gov (United States)

    Chatterjee, Tanaya; Sheikh, Irshad Ali; Chakravarty, Devlina; Chakrabarti, Pinak; Sarkar, Paramita; Saha, Tultul; Chakrabarti, Manoj K.; Hoque, Kazi Mirajul

    2015-01-01

    Cholera pathogenesis occurs due to synergistic pro-secretory effects of several toxins, such as cholera toxin (CTX) and Accessory cholera enterotoxin (Ace) secreted by Vibrio cholerae strains. Ace activates chloride channels stimulating chloride/bicarbonate transport that augments fluid secretion resulting in diarrhea. These channels have been targeted for drug development. However, lesser attention has been paid to the interaction of chloride channel modulators with bacterial toxins. Here we report the modulation of the structure/function of recombinant Ace by small molecule calcium-activated chloride channel (CaCC) inhibitors, namely CaCCinh-A01, digallic acid (DGA) and tannic acid. Biophysical studies indicate that the unfolding (induced by urea) free energy increases upon binding CaCCinh-A01 and DGA, compared to native Ace, whereas binding of tannic acid destabilizes the protein. Far-UV CD experiments revealed that the α-helical content of Ace-CaCCinh-A01 and Ace-DGA complexes increased relative to Ace. In contrast, binding to tannic acid had the opposite effect, indicating the loss of protein secondary structure. The modulation of Ace structure induced by CaCC inhibitors was also analyzed using docking and molecular dynamics (MD) simulation. Functional studies, performed using mouse ileal loops and Ussing chamber experiments, corroborate biophysical data, all pointing to the fact that tannic acid destabilizes Ace, inhibiting its function, whereas DGA stabilizes the toxin with enhanced fluid accumulation in mouse ileal loop. The efficacy of tannic acid in mouse model suggests that the targeted modulation of Ace structure may be of therapeutic benefit for gastrointestinal disorders. PMID:26540279

  13. Review of oral cholera vaccines: efficacy in young children

    Directory of Open Access Journals (Sweden)

    Masuet Aumatell C

    2011-09-01

    Full Text Available Cristina Masuet Aumatell1, JM Ramon Torrell1, Jane N Zuckerman21International Health Centre, Preventive Medicine Department, Bellvitge Hospital, L'Hospitalet de Llobregat, Barcelona, Spain; 2World Health Organization Collaborating Centre for Reference, Research and Training in Travel Medicine, University College London Medical School, London, UKBackground: Young children are one of the most vulnerable groups who may be infected with cholera. The following literature review of the efficacy of the currently available cholera vaccines provides a clear evidence base for the clinical administration of cholera vaccine, particularly in an epidemic situation.Aim: To assess the efficacy of oral cholera vaccines in preventing cases of cholera in young children.Methods: A systematic literature review was undertaken for the period 1983 to 2011 using PubMed and the search terms “oral cholera vaccines,” “children,” and “efficacy,” limited to “clinical trials” and “human studies”.Results: Oral cholera vaccine provides an acceptable level of protection in young children, with the level of protection being greater at 12 or 24 months following immunization.Conclusions: Children exposed to a potential risk of cholera are recommended to be vaccinated with an oral cholera vaccine, irrespective of whether its constituents include the B subunit.Keywords: efficacy, oral cholera vaccine, children

  14. Genome Sequence of Vibrio cholerae G4222, a South African Clinical Isolate

    OpenAIRE

    le Roux, Wouter J.; Chan, Wai Yin; De Maayer, Pieter; Venter, Stephanus N.

    2013-01-01

    Vibrio cholerae, a Gram-negative pathogen autochthonous to the aquatic environment, is the causative agent of cholera. Here, we report the complete genome sequence of V. cholerae G4222, a clinical isolate from South Africa.

  15. Biofilm formation and phenotypic variation enhance predation-driven persistence of Vibrio cholerae

    DEFF Research Database (Denmark)

    Matz, Carsten; McDougald, D.; Moreno, A.M.;

    2005-01-01

    Persistence of the opportunistic bacterial pathogen Vibrio cholerae in aquatic environments is the principal cause for seasonal occurrence of cholera epidemics. This causality has been explained by postulating that V. cholerae forms biofilms in association with animate and inanimate surfaces...

  16. Evaluation of Peru-15, a new live oral vaccine for cholera, in volunteers.

    Science.gov (United States)

    Sack, D A; Sack, R B; Shimko, J; Gomes, G; O'Sullivan, D; Metcalfe, K; Spriggs, D

    1997-07-01

    A new live oral cholera vaccine, Peru-15, was studied for safety, immunogenicity, and excretion in 2 groups of healthy volunteers. Twelve inpatient volunteers received freshly harvested vaccine in doses of either 10(7) or 10(9) cfu. Subsequently 50 outpatient volunteers received freeze-dried vaccine in doses of 10(8) or 10(9) cfu or placebo in a three-cell, double-masked, placebo-controlled trial. The strain was well tolerated at all dose levels, and it stimulated high levels of vibriocidal antibodies in most inpatient volunteers and in all outpatient volunteers. Although antitoxin responses were less frequent and of lower magnitude than the vibriocidal responses, antitoxin responses were seen in >60% of the outpatient volunteers. About 60% of the volunteers excreted the vaccine in their feces; however, fecal excretion did not correlate with serologic responses. It is concluded that Peru-15 is a safe and immunogenic oral vaccine for cholera.

  17. Sanitation in the time of cholera.

    Science.gov (United States)

    Misch, A

    1991-01-01

    Cholera, identified by violent diarrhea, cramps, vomiting, and dehydration, is spreading through Peru into Colombia, Ecuador, Child, and Brazil. Water contaminated with Vibrio cholerae is used for washing food and/or drinking thereby transmitting the disease. PAHO estimates 6 million people in South America may get cholera within the next 3 years. This cholera epidemic is the result of unsanitary conditions in which the urban poor in South America live. In fact, in Lima, Peru, 40% of the people do not have potable, piped water available. These individuals fetch their water from far away taps and private vendors both of which are not necessarily safe. In addition, 40% do not have access to a sewage system. Further, 80% of sick people in developing countries have a water related illness, be it transmitted by contaminated water or by insects and snails that reproduce in the water. Diarrhea is the most deadly of these conditions. Indeed every year 10-20 million children die from the effects of diarrhea which include malnutrition, dehydration, and shock. Yet 940 million people in developing countries have no access to safe water and 1.7 billion do not have a sanitary means of disposing of human wastes, despite the fact that the UN decreed the 1980s the International Drinking Water Supply and Sanitation Decade. Nevertheless UNICEF efforts did bring communal taps, odorless latrines, and/or pour flush toilets to 1.2 billion people. These types of sanitation costs $20-25/person whereas conventional sewers cost $350/person. Low technology supplied water averages $30/person compared to $200/person for piped water. Peru has spent $43 million on emergency medical care for cholera victims which could have provided low cost clean water and sanitation for almost 800,000 poor.

  18. Cytotoxic and Inflammatory Responses Induced by Outer Membrane Vesicle-Associated Biologically Active Proteases from Vibrio cholerae.

    Science.gov (United States)

    Mondal, Ayan; Tapader, Rima; Chatterjee, Nabendu Sekhar; Ghosh, Amit; Sinha, Ritam; Koley, Hemanta; Saha, Dhira Rani; Chakrabarti, Manoj K; Wai, Sun Nyunt; Pal, Amit

    2016-05-01

    Proteases in Vibrio cholerae have been shown to play a role in its pathogenesis. V. cholerae secretes Zn-dependent hemagglutinin protease (HAP) and calcium-dependent trypsin-like serine protease (VesC) by using the type II secretion system (TIISS). Our present studies demonstrated that these proteases are also secreted in association with outer membrane vesicles (OMVs) and transported to human intestinal epithelial cells in an active form. OMV-associated HAP induces dose-dependent apoptosis in Int407 cells and an enterotoxic response in the mouse ileal loop (MIL) assay, whereas OMV-associated VesC showed a hemorrhagic fluid response in the MIL assay, necrosis in Int407 cells, and an increased interleukin-8 (IL-8) response in T84 cells, which were significantly reduced in OMVs from VesC mutant strain. Our results also showed that serine protease VesC plays a role in intestinal colonization of V. cholerae strains in adult mice. In conclusion, our study shows that V. cholerae OMVs secrete biologically active proteases which may play a role in cytotoxic and inflammatory responses. PMID:26930702

  19. Epidemic cholera in Ecuador: multidrug-resistance and transmission by water and seafood.

    Science.gov (United States)

    Weber, J T; Mintz, E D; Cañizares, R; Semiglia, A; Gomez, I; Sempértegui, R; Dávila, A; Greene, K D; Puhr, N D; Cameron, D N

    1994-02-01

    To determine risk factors for cholera in an epidemic-disease area in South America, a case-control investigation was performed in Guayaquil, Ecuador, in July 1991. Residents > 5 years old who were hospitalized for treatment of acute, watery diarrhoea and two matched controls for each were interviewed regarding sources of water and food, and eating, drinking, and hygienic habits. Interviewers inspected homes of case-patients and controls to document water treatment, food-handling, and hygienic practices. Faecal specimens and shellfish were cultured for Vibrio cholerae O 1. Isolates were tested for susceptibility to a variety of antimicrobial agents. Drinking unboiled water (odds ratio [OR] = 4.0, confidence interval [CI] = 1.8-7.5), drinking a beverage from a street vendor (OR = 2.8, CI = 1.3-5.9), eating raw seafood (OR = 3.4, CI = 1.4-11.5), and eating cooked crab (OR = 5.1, CI = 1.4-19.2) were associated with illness. Always boiling drinking water at home (OR = 0.5, CI = 0.2-0.9) was protective against illness. The presence of soap in either the kitchen (OR = 0.3, CI = 0.2-0.8) or bathroom (OR = 0.4, CI = 0.2-0.9) at home was also protective. V. cholerae O 1 was recovered from a pooled sample of a bivalve mollusc and from 68% of stool samples from case-patients. Thirty-six percent of the isolates from stool specimens were resistant to multiple antimicrobial agents. Specific prevention measures may prevent transmission through these vehicles in the future. The appearance of antimicrobial resistance suggests the need for changes in current methods of prevention and treatment. PMID:8119348

  20. Introduction on molecular biological techniques for Vibrio cholerae%霍乱弧菌分子生物学检测技术介绍

    Institute of Scientific and Technical Information of China (English)

    熊长辉

    2012-01-01

    随着医学分子生物学的迅速发展,大量的分子生物学技术被用于霍乱弧菌的研究,为霍乱弧菌快速检测及分型提供了重要依据,也进一步从分子水平阐明了霍乱弧菌的变异和不同菌株之间的遗传关系,以及霍乱疫情的溯源、菌株类型和流行性质.%Molecular biological techniques provide an important basis for the rapid detection of Vibrio cholerae with the rapid development of Medical Molecular Biology, increasing number of molecular biology techniques were used to detect Vibrio cholerae variation and genetic relationships between different strains, the origin, strain type and nature of the epidemic cholera.

  1. SURVEY ON V. CHOLERAE, V. VULNIFICUS AND V. PARAHAEMOLYTICUS IN BIVALVE MOLLUSCS OF THE ADRIATIC SEA AND PROPOSAL OF AN ANALYTICAL PROTOCOL

    Directory of Open Access Journals (Sweden)

    M.L. Valeri

    2009-06-01

    Full Text Available Bivalve molluscs from Adriatic sea were analyzed for V. parahaemolyticus, V. cholerae e V. vulnificus presence. The isolates on TCBS Agar and m-CPC Agar were selected on the basis of a new biochemical screening, that showed a good performance, because among 2344 strains from primary culture only 237 (10% were presumptively assigned to the species of interest. The PCR analyses was performed for the target genes toxR hlyA, ctxA, tcpI (V. cholerae, toxR, tl, tdh, trh (V. parahaemolyticus, vvhA and viuB (V. vulnificus. Among the 9 strains confirmed to belong to V. parahaemolyticus specie, 6 were sucrose positive. On 215 samples of molluscs only 5 resulted positive for V. parahaemolyticus being toxR+, tl+, although non pathogenic (tdh-, trh-, and none for V. cholerae e V. vulnificus.

  2. The role of Vibrio cholerae genotyping in Africa.

    Science.gov (United States)

    De, Rituparna; Ghosh, Jayeeta Banerjee; Sen Gupta, Sourav; Takeda, Yoshifumi; Nair, G Balakrish

    2013-11-01

    Toxigenic Vibrio cholerae, the causative agent of the disease cholera, is prevalent in the African continent from the 1970s when the seventh pandemic spread from Asia to Africa. In the past decade, cholera has caused devastating outbreaks in much of Africa, illustrated by the recent cholera epidemics in Zimbabwe and regions of central Africa. Given the extent of cholera in Africa, a robust and efficient surveillance system should be in place to prevent and control the disease in this continent. Such a surveillance system would be greatly bolstered by use of molecular typing techniques to identify genetic subtypes. In this review, we highlight the role that modern molecular typing techniques can play in tracking and aborting the spread of cholera. PMID:24101642

  3. Detection and identification of enterohemorrhagic Escherichia coli O157:H7 and Vibrio cholerae O139 using oligonucleotide microarray

    Directory of Open Access Journals (Sweden)

    Zhang Zheng

    2007-12-01

    Full Text Available Abstract Background The rapid and accurate detection and identification of the new subtype of the pathogens is crucial for diagnosis, treatment and control of the contagious disease outbreak. Here, in this study, an approach to detect and identify Escherichia coli O157:H7 and Vibrio cholerae O139 was established using oligonucleotide microarray. We coupled multiplex PCR with oligonucleotide microarray to construct an assay suitable for simultaneous identification of two subtypes of the pathogens. Results The stx1, stx2 gene and uidA gene having the specific mutant spot were chosen as the targets for Escherichia coli O157:H7, and meanwhile the ctxA, tcpA, and LPSgt gene for Vibrio cholerae O139. The oligonucleotide microarray was composed of eight probes including negative control and positive control from 16S rDNA gene. The six primers were designed to amplify target fragments in two triplex PCR, and then hybridized with oligonucleotide microarray. An internal control would be to run a PCR reaction in parallel. Multiplex PCR did not produce any non-specific amplicons when 149 related species or genera of standard bacteria were tested (100% specificity. In addition, Escherichia coli O157:H7 and Escherichia coli O157:non-H7, Vibrio cholerae O139 and Vibrio cholerae O1 had been discriminated respectively. Using recombinant plasmid and target pathogens, we were able to detect positive hybridization signals with 102 copies/μL and 103 cfu/mL per reaction. Conclusion The DNA microarray assay reported here could detect and identify Escherichia coli O157:H7 and Vibrio cholerae O139, and furthermore the subtype was distinguished. This assay was a specific and sensitive tool for simultaneous detection and identification of the new subtype of two pathogens causing diarrhea in human.

  4. Cholera in Portugal, 1974. II. Transmission by bottled mineral water.

    Science.gov (United States)

    Blake, P A; Rosenberg, M L; Florencia, J; Costa, J B; do Prado Quintino, L; Gangarosa, E J

    1977-04-01

    During a cholera epidemic, Vibrio cholerae was isolated from two springs which supplied mineral water to a spa and to a commercial water bottling plant. Epidemiologic investigation found that cholera attack rates were 10-fold greater among visitors to the spa than among non-visitors. A subsequent matched-pair case-control study which excluded persons who had visted the spa showed that a history of consumption of the bottled non-carbonated water was significantly more common among bacteriologically confirmed cholera cases than among paired controls.

  5. Comparative microscopy study of Vibrio cholerae flagella

    Science.gov (United States)

    Konnov, Nikolai P.; Baiburin, Vil B.; Zadnova, Svetlana P.; Volkov, Uryi P.

    1999-06-01

    A fine structure of bacteria flagella is an important problem of molecular cell biology. Bacteria flagella are the self-assembled structures that allow to use the flagellum protein in a number of biotechnological applications. However, at present, there is a little information about high resolution scanning probe microscopy study of flagellum structure, in particular, about investigation of Vibrio cholerae flagella. In our lab have been carried out the high resolution comparative investigation of V. cholerae flagella by means of various microscopes: tunneling (STM), scanning force (SFM) and electron transmission. As a scanning probe microscope is used designed in our lab versatile SPM with replaceable measuring heads. Bacteria were grown, fixed and treated according to the conventional techniques. For STM investigations samples were covered with Pt/Ir thin films by rotated vacuum evaporation, in SFM investigations were used uncovered samples. Electron microscopy of the negatively stained bacteria was used as a test procedure.

  6. Comparative analysis of different oral approaches to treat Vibrio cholerae infection in adult mice.

    Science.gov (United States)

    Jaiswal, Abhishek; Koley, Hemanta; Mitra, Soma; Saha, Dhira Rani; Sarkar, Banwarilal

    2014-05-01

    In this study, we have established an oral phage cocktail therapy in adult mice model and also performed a comparative analysis between phage cocktail, antibiotic and oral rehydration treatment for orally developed Vibrio cholerae infection. Four groups of mice were orally infected with Vibrio cholerae MAK 757 strain. Phage cocktail and antibiotic treated groups received 1×10(8) plaque forming unit/ml (once a daily) and 40mg/kg (once a daily) as an oral dose respectively for consecutive three days after bacterial infection. In case of oral rehydration group, the solution was supplied after bacterial infection mixed with the drinking water. To evaluate the better and safer approach of treatment, tissue and serum samples were collected. Here, phage cocktail treated mice reduced the log10 numbers of colony per gram by 3log10 (p0.05). Besides, it was evident that antibiotic and phage cocktail treated group had a gradual decrease in both IL-6 and TNF-α level for 3 days (pVibrio cholerae infection.

  7. Mapping to Support Fine Scale Epidemiological Cholera Investigations: A Case Study of Spatial Video in Haiti.

    Science.gov (United States)

    Curtis, Andrew; Blackburn, Jason K; Smiley, Sarah L; Yen, Minmin; Camilli, Andrew; Alam, Meer Taifur; Ali, Afsar; Morris, J Glenn

    2016-02-03

    The cartographic challenge in many developing world environments suffering a high disease burden is a lack of granular environmental covariates suitable for modeling disease outcomes. As a result, epidemiological questions, such as how disease diffuses at intra urban scales are extremely difficult to answer. This paper presents a novel geospatial methodology, spatial video, which can be used to collect and map environmental covariates, while also supporting field epidemiology. An example of epidemic cholera in a coastal town of Haiti is used to illustrate the potential of this new method. Water risks from a 2012 spatial video collection are used to guide a 2014 survey, which concurrently included the collection of water samples, two of which resulted in positive lab results "of interest" (bacteriophage specific for clinical cholera strains) to the current cholera situation. By overlaying sample sites on 2012 water risk maps, a further fifteen proposed water sample locations are suggested. These resulted in a third spatial video survey and an additional "of interest" positive water sample. A potential spatial connection between the "of interest" water samples is suggested. The paper concludes with how spatial video can be an integral part of future fine-scale epidemiological investigations for different pathogens.

  8. Mapping to Support Fine Scale Epidemiological Cholera Investigations: A Case Study of Spatial Video in Haiti

    Directory of Open Access Journals (Sweden)

    Andrew Curtis

    2016-02-01

    Full Text Available The cartographic challenge in many developing world environments suffering a high disease burden is a lack of granular environmental covariates suitable for modeling disease outcomes. As a result, epidemiological questions, such as how disease diffuses at intra urban scales are extremely difficult to answer. This paper presents a novel geospatial methodology, spatial video, which can be used to collect and map environmental covariates, while also supporting field epidemiology. An example of epidemic cholera in a coastal town of Haiti is used to illustrate the potential of this new method. Water risks from a 2012 spatial video collection are used to guide a 2014 survey, which concurrently included the collection of water samples, two of which resulted in positive lab results “of interest” (bacteriophage specific for clinical cholera strains to the current cholera situation. By overlaying sample sites on 2012 water risk maps, a further fifteen proposed water sample locations are suggested. These resulted in a third spatial video survey and an additional “of interest” positive water sample. A potential spatial connection between the “of interest” water samples is suggested. The paper concludes with how spatial video can be an integral part of future fine-scale epidemiological investigations for different pathogens.

  9. An analysis of the V1 and V2 regions of Vibrio cholerae and Vibrio mimicus 16S rRNA.

    Science.gov (United States)

    Coelho, A; Momen, H; Vicente, A C; Salles, C A

    1994-02-01

    The V1 and V2 variable regions of the 16S rRNA gene of three strains of V. cholerae and one strain of V. mimicus were amplified by PCR. Fragments containing both regions were cloned into M13mp18 using Smal and sequenced by the dideoxy method. The 263-bp sequence from a strain isolated during the 1991 cholera outbreak in Brazil was deposited in Genbank under the accession number L05178. Except for an extra G in one of the strains, the three V. cholerae sequences were identical. The V. mimicus sequence was very similar, with only two substitutions. We compared these sequences with the Vibrio 16S rRNA sequences described by Dorsch et al. in 1992. It was noted that the V1 region, including helix 6 and its associated loop, comprised two different sizes and sequences in the various Vibrio species. While V. cholerae, V. mimicus, V. vulnificus, V. anguillarum and V. diazotrophicus had a 46-nucleotide V1, other species such as V. parahaemolyticus, V. proteolyticus, V. alginolyticus, V. campbellii and V. hollisae had longer 54- or 55-nucleotide regions, with a different consensus sequence. The phylogeny of Vibrio was analysed using the sequenced region and its equivalent in other species, by means of the "Phylip" software package. Species with a short helix 6 were grouped together, as were species with a long helix. Dorsh et al.'s analysis is discussed in relation to this "helix 6 split".

  10. CHOLERA EL-TOR EN IRAN

    Directory of Open Access Journals (Sweden)

    M. Ghodssi

    1969-01-01

    Full Text Available The bacteriological analysis shows that we have been confronted with the ELTor type, and only that type, until the end of the epidemic.The clinical study presents the symptoms of the real cholera with all its grievous consequences.The epidemiological supertnisicn stales that the El..Tor cholera is not agressiveat all in town areas whereas it presents its usual aspect in country areas, because of a lack of hygiene. there.That disease can be completely cured if the balance between the electrolytesis quickly restored.The disease was all the more dreadful since it came as a surprise and spread from one province to the other.L'examen bacteriologique montrc qu'il s'agit du type EI_ Tor et uniquement du meme type jusqu'a la fin de l'epldemie.La surveillance epidemiologique constate que Ie cholera EI_Tor n'cst nullement agressif dans Ie milieu urbain; mais qu'H revet l'aspect classique dans les milieuxruraux, depourvus d'hyglene.La maludic est totalement guerissablc a condition que l'equilibre des electrolytes so it rapidement retabli. L'evenement a tHe maladie se repandit d'une12

  11. The highly conserved bacterial RNase YbeY is essential in Vibrio cholerae, playing a critical role in virulence, stress regulation, and RNA processing.

    Science.gov (United States)

    Vercruysse, Maarten; Köhrer, Caroline; Davies, Bryan W; Arnold, Markus F F; Mekalanos, John J; RajBhandary, Uttam L; Walker, Graham C

    2014-06-01

    YbeY, a highly conserved protein, is an RNase in E. coli and plays key roles in both processing of the critical 3' end of 16 S rRNA and in 70 S ribosome quality control under stress. These central roles account for YbeY's inclusion in the postulated minimal bacterial genome. However, YbeY is not essential in E. coli although loss of ybeY severely sensitizes it to multiple physiological stresses. Here, we show that YbeY is an essential endoribonuclease in Vibrio cholerae and is crucial for virulence, stress regulation, RNA processing and ribosome quality control, and is part of a core set of RNases essential in most representative pathogens. To understand its function, we analyzed the rRNA and ribosome profiles of a V. cholerae strain partially depleted for YbeY and other RNase mutants associated with 16 S rRNA processing; our results demonstrate that YbeY is also crucial for 16 S rRNA 3' end maturation in V. cholerae and that its depletion impedes subunit assembly into 70 S ribosomes. YbeY's importance to V. cholerae pathogenesis was demonstrated by the complete loss of mice colonization and biofilm formation, reduced cholera toxin production, and altered expression levels of virulence-associated small RNAs of a V. cholerae strain partially depleted for YbeY. Notably, the ybeY genes of several distantly related pathogens can fully complement an E. coli ΔybeY strain under various stress conditions, demonstrating the high conservation of YbeY's activity in stress regulation. Taken together, this work provides the first comprehensive exploration of YbeY's physiological role in a human pathogen, showing its conserved function across species in essential cellular processes.

  12. Authentic display of a cholera toxin epitope by chimeric type 1 fimbriae: effects of insert position and host background

    DEFF Research Database (Denmark)

    Stentebjerg-Olesen, B; Pallesen, L; Jensen, LB;

    1997-01-01

    with respect to host background in three different Escherichia coli strains, i.e. an isogenic set of K-12 strains, differing in the presence of an indigenous fim gene cluster, as well as a wild-type isolate. Immunization of rabbits with purified chimeric fimbriae resulted in serum which specifically recognized......The potential of the major structural protein of type 1 fimbriae as a display system for heterologous sequences was tested. As a reporter-epitope, a heterologous sequence mimicking a neutralizing epitope of the cholera toxin B chain was inserted, in one or two copies, into four different positions....... Several of the chosen positions seemed amenable even for large foreign inserts; the chimeric proteins were exposed on the bacterial surface and the cholera toxin epitope was authentically displayed, i.e. it was recognized on bacteria by specific antiserum. Display of chimeric fimbriae was tested...

  13. FtsK-dependent dimer resolution on multiple chromosomes in the pathogen Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    Marie-Eve Val

    Full Text Available Unlike most bacteria, Vibrio cholerae harbors two distinct, nonhomologous circular chromosomes (chromosome I and II. Many features of chromosome II are plasmid-like, which raised questions concerning its chromosomal nature. Plasmid replication and segregation are generally not coordinated with the bacterial cell cycle, further calling into question the mechanisms ensuring the synchronous management of chromosome I and II. Maintenance of circular replicons requires the resolution of dimers created by homologous recombination events. In Escherichia coli, chromosome dimers are resolved by the addition of a crossover at a specific site, dif, by two tyrosine recombinases, XerC and XerD. The process is coordinated with cell division through the activity of a DNA translocase, FtsK. Many E. coli plasmids also use XerCD for dimer resolution. However, the process is FtsK-independent. The two chromosomes of the V. cholerae N16961 strain carry divergent dimer resolution sites, dif1 and dif2. Here, we show that V. cholerae FtsK controls the addition of a crossover at dif1 and dif2 by a common pair of Xer recombinases. In addition, we show that specific DNA motifs dictate its orientation of translocation, the distribution of these motifs on chromosome I and chromosome II supporting the idea that FtsK translocation serves to bring together the resolution sites carried by a dimer at the time of cell division. Taken together, these results suggest that the same FtsK-dependent mechanism coordinates dimer resolution with cell division for each of the two V. cholerae chromosomes. Chromosome II dimer resolution thus stands as a bona fide chromosomal process.

  14. Central role of the Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR) in sodium bioenergetics of Vibrio cholerae.

    Science.gov (United States)

    Steuber, Julia; Halang, Petra; Vorburger, Thomas; Steffen, Wojtek; Vohl, Georg; Fritz, Günter

    2014-12-01

    Vibrio cholerae is a Gram-negative bacterium that lives in brackish or sea water environments. Strains of V. cholerae carrying the pathogenicity islands infect the human gut and cause the fatal disease cholera. Vibrio cholerae maintains a Na(+) gradient at its cytoplasmic membrane that drives substrate uptake, motility, and efflux of antibiotics. Here, we summarize the major Na(+)-dependent transport processes and describe the central role of the Na(+)-translocating NADH:quinone oxidoreductase (Na(+)-NQR), a primary Na(+) pump, in maintaining a Na(+)-motive force. The Na(+)-NQR is a membrane protein complex with a mass of about 220 kDa that couples the exergonic oxidation of NADH to the transport of Na(+) across the cytoplasmic membrane. We describe the molecular architecture of this respiratory complex and summarize the findings how electron transport might be coupled to Na(+)-translocation. Moreover, recent advances in the determination of the three-dimensional structure of this complex are reported.

  15. Differential RNA-seq of Vibrio cholerae identifies the VqmR small RNA as a regulator of biofilm formation.

    Science.gov (United States)

    Papenfort, Kai; Förstner, Konrad U; Cong, Jian-Ping; Sharma, Cynthia M; Bassler, Bonnie L

    2015-02-17

    Quorum sensing (QS) is a process of cell-to-cell communication that enables bacteria to transition between individual and collective lifestyles. QS controls virulence and biofilm formation in Vibrio cholerae, the causative agent of cholera disease. Differential RNA sequencing (RNA-seq) of wild-type V. cholerae and a locked low-cell-density QS-mutant strain identified 7,240 transcriptional start sites with ∼ 47% initiated in the antisense direction. A total of 107 of the transcripts do not appear to encode proteins, suggesting they specify regulatory RNAs. We focused on one such transcript that we name VqmR. vqmR is located upstream of the vqmA gene encoding a DNA-binding transcription factor. Mutagenesis and microarray analyses demonstrate that VqmA activates vqmR transcription, that vqmR encodes a regulatory RNA, and VqmR directly controls at least eight mRNA targets including the rtx (repeats in toxin) toxin genes and the vpsT transcriptional regulator of biofilm production. We show that VqmR inhibits biofilm formation through repression of vpsT. Together, these data provide to our knowledege the first global annotation of the transcriptional start sites in V. cholerae and highlight the importance of posttranscriptional regulation for collective behaviors in this human pathogen.

  16. Vibrio cholerae O139 in Thailand in 1994.

    OpenAIRE

    Bodhidatta, L.; Echeverria, P; Hoge, C W; Pitarangsi, C; Serichantalergs, O; Henprasert-Tae, N.; Harikul, S.; Kitpoka, P.

    1995-01-01

    Vibrio cholerae O139 first appeared in India and Bangladesh in 1992. Surveillance for O139 was started at three hospitals in Thailand in 1993. By 1994 all three hospitals surveyed in Thailand had experienced an increase in Vibrio cholerae O139 infections.

  17. Synthesis of protein in intestinal cells exposed to cholera toxin

    International Nuclear Information System (INIS)

    The mechanism by which cyclic adenosine monophosphate (AMP), formed by intestinal epithelial cells in response to cholera toxin, ultimately results in alterations in water and electrolyte transport is poorly understood. Several studies have indicated that inhibitors of transcription or translation block much of the transport of ions and water in the intestine and edema formation in tissue elicited by cholera toxin. Data presented in this study confirmed the inhibitory effects of cycloheximide on cholera toxin-induced fluid accumulation in the rabbit intestinal loop model. Neither cycloheximide nor actinomycin D altered the amount of cyclic AMP that accumulated in intestinal cells and Chinese hamster ovary cells exposed to cholera toxin. An increase in [3H] leucine incorporation was readily demonstrable in intestinal epithelial cells from rabbits challenged with Vibrio cholerae. Similarly, intestinal epithelial cells incubated with cholera toxin for 4 hr synthesized substantially more protein than controls as determined by relative incorporation of [35S] methionine. Most of the new protein synthesized in response to cholera toxin was membrane associated and of high molecular weight. The possible significance of the toxin-induced protein relative to cholera pathogenesis was discussed

  18. Understanding the Hydrology of Cholera in South Asia

    Science.gov (United States)

    Akanda, A. S.; Jutla, A. S.; Islam, S.

    2007-12-01

    Cholera is an acute waterborne illness caused by the bacterium Vibrio cholerae. The disease remains a major public health issue in several regions of the developing world, mainly in coastal areas around the tropics. Cholera incidences have been historically linked to climate variables and more recently with El Nino-Southern Oscillation. The occurrence of cholera shows bi-annual seasonal peaks and strong inter-annual variability in the Ganges basin region of South Asia. However, the role of hydrologic variables in the seasonal patterns of cholera epidemics is less understood. Preliminary results suggest that a unique combination of increasing water temperature and higher salinity in the coastal zone during the low flow season provide the situation amenable to the first outbreak of cholera in the spring season. Other major factors contributing to the subsequent spread of the disease are sea surface height, monsoon precipitation, and coastal phytoplankton concentration. We will further examine the lag periods between the dominant environmental variables and cholera incidences to understand the seasonal dynamics of cholera in South Asia.

  19. Investigation of household contamination of Vibrio cholerae in Bangladesh

    DEFF Research Database (Denmark)

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

    The role of in-house transmission on the incidence of Vibrio cholerae, the deadly waterborne pathogen, is still not developed. The aim of the current study was to investigate possible contamination routes in household domain for effective cholera control in Bangladesh. To examine the prevalence...... and water supply may be the reason behind this relatively high presence of virulence factors in food plates and water pots. Direct exposure routes of disease transmission should be a major consideration in cholera prevention policies. Investigation of household contamination of Vibrio cholerae in Bangladesh........ Available from: https://www.researchgate.net/publication/305215719_Investigation_of_household_contamination_of_Vibrio_cholerae_in_Bangladesh [accessed Oct 14, 2016]....

  20. USE OF MODIFIED CAMP TEST FOR PRELIMINARY NONSEROLOGIC IDENTIFICATION OF VIBRIO CHOLERAE IN STOOL SPECIMENS

    Directory of Open Access Journals (Sweden)

    Murad Lesmana

    2012-09-01

    Full Text Available Suatu modifikasi uji CAMP digunakan bersama dengan reaksi biokimiawi untuk identifikasi Vibrio cholerae pada sampel klinis. Dari 579 usap dubur penderita diare, 92 (16% memberikan hasil isolasi V. cholerae 01 biotipe El Tor dan 34 (6% V. cholerae non-01. Semua isolat V. cholerae 01 El Tor menunjukkan reaksi CAMP positif kuat dengan gambaran hemolisis sinergistik lengkap berbentuk sosis; sedangkan V. cholerae non-01 memberikan reaksi CAMP yang sempit dengan pola hemolisis menyerupai bulan sabit. Hasil uji CAMP yang dilakukan bersama dengan reaksi biokimiawi sesuai dengan metode biakan konvensional yang menyertakan tes aglutinasi dengan antiserum V. cholerae 01 untuk mengidentifikasi V. cholerae.

  1. Spatially selective colonization of the arthropod intestine through activation of Vibrio cholerae biofilm formation

    OpenAIRE

    Purdy, Alexandra E.; Watnick, Paula I.

    2011-01-01

    Vibrio cholerae is an estuarine bacterium and the human pathogen responsible for the diarrheal disease cholera. In the environment, arthropods are proposed to be carriers and reservoirs of V. cholerae. However, the molecular basis of the association between V. cholerae and viable arthropods has not been elucidated previously. Here, we show that the V. cholerae Vibrio polysaccharide (VPS)-dependent biofilm is highly activated upon entry into the arthropod intestine and is specifically required...

  2. A Metalloprotease Secreted by the Type II Secretion System Links Vibrio cholerae with Collagen

    OpenAIRE

    Park, Bo R.; Ryszard A Zielke; Wierzbicki, Igor H.; Mitchell, Kristie C.; Withey, Jeffrey H.; Sikora, Aleksandra E.

    2015-01-01

    Vibrio cholerae is autochthonous to various aquatic niches and is the etiological agent of the life-threatening diarrheal disease cholera. The persistence of V. cholerae in natural habitats is a crucial factor in the epidemiology of cholera. In contrast to the well-studied V. cholerae-chitin connection, scarce information is available about the factors employed by the bacteria for the interaction with collagens. Collagens might serve as biologically relevant substrates, because they are the m...

  3. Isolation and characterization of Vibrio cholerae isolates from seafood in Hat Yai City, Songkhla, Thailand.

    Science.gov (United States)

    Preeprem, Sutima; Mittraparp-arthorn, Pimonsri; Bhoopong, Phuangthip; Vuddhakul, Varaporn

    2014-11-01

    Seafood has been identified as an important source of Vibrio cholerae in Thailand, especially in the Southern coastal region. In this study, we isolated and characterized V. cholerae from seafood obtained from several markets in Hat Yai city, Southern Thailand. A total of 100 V. cholerae isolates were obtained from 55 of 125 seafood samples. The dominant serotype was non-O1/non-O139. Polymerase chain reaction (PCR) analysis was used to detect the presence of pathogenesis-related genes. The stn/sto and hlyA El Tor virulence genes were detected in 20% and 96% of the isolates, respectively. None of the isolates were positive for the ctxA, tcpA, zot, and ace genes. Only 6% of the isolates carried the T3SS gene (vcsV2); however, the majority of the isolates (96%) carried the T6SS gene (vasH). Representative isolates (n=35) that exhibited various virulence gene patterns were randomly selected and analyzed for their hemolytic activity, antibiotic susceptibility, biofilm formation, and genotype. Hemolytic activity using sheep red blood cells was detected in only one of the hlyA-negative isolates. Apart from ampicillin, all isolates were pansusceptible to five test antibiotics. Biofilm production was observed in most of the isolates, and there was no difference in the presence of a biofilm between the smooth and rugose isolates. Using the enterobacterial repetitive intergenic consensus-PCR method, clonal relationships were observed among the isolates that exhibited identical virulence gene patterns.

  4. Host cell contact induces expression of virulence factors and VieA, a cyclic di-GMP phosphodiesterase, in Vibrio cholerae.

    Science.gov (United States)

    Dey, Amit K; Bhagat, Abha; Chowdhury, Rukhsana

    2013-05-01

    Vibrio cholerae, a noninvasive bacterium, colonizes the intestinal epithelium and secretes cholera toxin (CT), a potent enterotoxin that causes the severe fluid loss characteristic of the disease cholera. In this study, we demonstrate that adherence of V. cholerae to the intestinal epithelial cell line INT 407 strongly induces the expression of the major virulence genes ctxAB and tcpA and the virulence regulatory gene toxT. No induction of toxR and tcpP, which encode transcriptional activators of toxT, was observed in adhered bacteria, and the adherence-dependent upregulation of toxT expression was independent of ToxR and TcpP. A sharp increase in the expression of the vieA gene, which encodes a cyclic di-GMP (c-di-GMP) phosphodiesterase, was observed in INT 407-adhered V. cholerae immediately after infection. Induction of toxT, ctxAB, and tcpA in INT 407-adhered vieA mutant strain O395 ΔvieA was consistently lower than in the parent strain, although no effect was observed in unadhered bacteria, suggesting that VieA has a role in the upregulation of toxT expression specifically in host cell-adhered V. cholerae. Furthermore, though VieA has both a DNA binding helix-turn-helix domain and an EAL domain conferring c-di-GMP phosphodiesterase activity, the c-di-GMP phosphodiesterase activity of VieA is necessary and sufficient for the upregulation of toxT expression.

  5. A factor converting viable but nonculturable Vibrio cholerae to a culturable state in eukaryotic cells is a human catalase.

    Science.gov (United States)

    Senoh, Mitsutoshi; Hamabata, Takashi; Takeda, Yoshifumi

    2015-08-01

    In our previous work, we demonstrated that viable but nonculturable (VBNC) Vibrio cholerae O1 and O139 were converted to culturable by coculture with eukaryotic cells. Furthermore, we isolated a factor converting VBNC V. cholerae to culturable (FCVC) from a eukaryotic cell line, HT-29. In this study, we purified FCVC by successive column chromatographies comprising UNO Q-6 anion exchange, Bio-Scale CHT2-1 hydroxyapatite, and Superdex 200 10/300 GL. Homogeneity of the purified FCVC was demonstrated by SDS-PAGE. Nano-LC MS/MS analysis showed that the purified FCVC was a human catalase. An experiment of RNAi knockdown of catalase mRNA from HT-29 cells and treatment of the purified FCVC with a catalase inhibitor, 3-amino-1,2,4-triazole confirmed that the FCVC was a catalase. A possible role of the catalase in converting a VBNC V. cholerae to a culturable state in the human intestine is discussed.

  6. Molecular tools in understanding the evolution of Vibrio cholerae.

    Science.gov (United States)

    Rahaman, Md Habibur; Islam, Tarequl; Colwell, Rita R; Alam, Munirul

    2015-01-01

    Vibrio cholerae, the etiological agent of cholera, has been a scourge for centuries. Cholera remains a serious health threat for developing countries and has been responsible for millions of deaths globally over the past 200 years. Identification of V. cholerae has been accomplished using a variety of methods, ranging from phenotypic strategies to DNA based molecular typing and currently whole genomic approaches. This array of methods has been adopted in epidemiological investigations, either singly or in the aggregate, and more recently for evolutionary analyses of V. cholerae. Because the new technologies have been developed at an ever increasing pace, this review of the range of fingerprinting strategies, their relative advantages and limitations, and cholera case studies was undertaken. The task was challenging, considering the vast amount of the information available. To assist the study, key references representative of several areas of research are provided with the intent to provide readers with a comprehensive view of recent advances in the molecular epidemiology of V. cholerae. Suggestions for ways to obviate many of the current limitations of typing techniques are also provided. In summary, a comparative report has been prepared that includes the range from traditional typing to whole genomic strategies.

  7. Vibrio cholerae Response Regulator VxrB Controls Colonization and Regulates the Type VI Secretion System.

    Directory of Open Access Journals (Sweden)

    Andrew T Cheng

    2015-05-01

    Full Text Available Two-component signal transduction systems (TCS are used by bacteria to sense and respond to their environment. TCS are typically composed of a sensor histidine kinase (HK and a response regulator (RR. The Vibrio cholerae genome encodes 52 RR, but the role of these RRs in V. cholerae pathogenesis is largely unknown. To identify RRs that control V. cholerae colonization, in-frame deletions of each RR were generated and the resulting mutants analyzed using an infant mouse intestine colonization assay. We found that 12 of the 52 RR were involved in intestinal colonization. Mutants lacking one previously uncharacterized RR, VCA0566 (renamed VxrB, displayed a significant colonization defect. Further experiments showed that VxrB phosphorylation state on the predicted conserved aspartate contributes to intestine colonization. The VxrB regulon was determined using whole genome expression analysis. It consists of several genes, including those genes that create the type VI secretion system (T6SS. We determined that VxrB is required for T6SS expression using several in vitro assays and bacterial killing assays, and furthermore that the T6SS is required for intestinal colonization. vxrB is encoded in a four gene operon and the other vxr operon members also modulate intestinal colonization. Lastly, though ΔvxrB exhibited a defect in single-strain intestinal colonization, the ΔvxrB strain did not show any in vitro growth defect. Overall, our work revealed that a small set of RRs is required for intestinal colonization and one of these regulators, VxrB affects colonization at least in part through its regulation of T6SS genes.

  8. Vibrio cholerae Response Regulator VxrB Controls Colonization and Regulates the Type VI Secretion System.

    Science.gov (United States)

    Cheng, Andrew T; Ottemann, Karen M; Yildiz, Fitnat H

    2015-05-01

    Two-component signal transduction systems (TCS) are used by bacteria to sense and respond to their environment. TCS are typically composed of a sensor histidine kinase (HK) and a response regulator (RR). The Vibrio cholerae genome encodes 52 RR, but the role of these RRs in V. cholerae pathogenesis is largely unknown. To identify RRs that control V. cholerae colonization, in-frame deletions of each RR were generated and the resulting mutants analyzed using an infant mouse intestine colonization assay. We found that 12 of the 52 RR were involved in intestinal colonization. Mutants lacking one previously uncharacterized RR, VCA0566 (renamed VxrB), displayed a significant colonization defect. Further experiments showed that VxrB phosphorylation state on the predicted conserved aspartate contributes to intestine colonization. The VxrB regulon was determined using whole genome expression analysis. It consists of several genes, including those genes that create the type VI secretion system (T6SS). We determined that VxrB is required for T6SS expression using several in vitro assays and bacterial killing assays, and furthermore that the T6SS is required for intestinal colonization. vxrB is encoded in a four gene operon and the other vxr operon members also modulate intestinal colonization. Lastly, though ΔvxrB exhibited a defect in single-strain intestinal colonization, the ΔvxrB strain did not show any in vitro growth defect. Overall, our work revealed that a small set of RRs is required for intestinal colonization and one of these regulators, VxrB affects colonization at least in part through its regulation of T6SS genes.

  9. Cholera outbreak in Senegal in 2005: was climate a factor?

    Directory of Open Access Journals (Sweden)

    Guillaume Constantin de Magny

    Full Text Available Cholera is an acute diarrheal illness caused by Vibrio cholerae and occurs as widespread epidemics in Africa. In 2005, there were 31,719 cholera cases, with 458 deaths in the Republic of Senegal. We retrospectively investigated the climate origin of the devastating floods in mid-August 2005, in the Dakar Region of Senegal and the subsequent outbreak of cholera along with the pattern of cholera outbreaks in three other regions of that country. We compared rainfall patterns between 2002 and 2005 and the relationship between the sea surface temperature (SST gradient in the tropical Atlantic Ocean and precipitation over Senegal for 2005. Results showed a specific pattern of rainfall throughout the Dakar region during August, 2005, and the associated rainfall anomaly coincided with an exacerbation of the cholera epidemic. Comparison of rainfall and epidemiological patterns revealed that the temporal dynamics of precipitation, which was abrupt and heavy, was presumably the determining factor. Analysis of the SST gradient showed that the Atlantic Ocean SST variability in 2005 differed from that of 2002 to 2004, a result of a prominent Atlantic meridional mode. The influence of this intense precipitation on cholera transmission over a densely populated and crowded region was detectable for both Dakar and Thiès, Senegal. Thus, high resolution rainfall forecasts at subseasonal time scales should provide a way forward for an early warning system in Africa for cholera and, thereby, trigger epidemic preparedness. Clearly, attention must be paid to both natural and human induced environmental factors to devise appropriate action to prevent cholera and other waterborne disease epidemics in the region.

  10. Cholera Outbreak in Senegal in 2005: Was Climate a Factor?

    Science.gov (United States)

    Constantin de Magny, Guillaume; Thiaw, Wassila; Kumar, Vadlamani; Manga, Noël M.; Diop, Bernard M.; Gueye, Lamine; Kamara, Mamina; Roche, Benjamin; Murtugudde, Raghu; Colwell, Rita R.

    2012-01-01

    Cholera is an acute diarrheal illness caused by Vibrio cholerae and occurs as widespread epidemics in Africa. In 2005, there were 31,719 cholera cases, with 458 deaths in the Republic of Senegal. We retrospectively investigated the climate origin of the devastating floods in mid-August 2005, in the Dakar Region of Senegal and the subsequent outbreak of cholera along with the pattern of cholera outbreaks in three other regions of that country. We compared rainfall patterns between 2002 and 2005 and the relationship between the sea surface temperature (SST) gradient in the tropical Atlantic Ocean and precipitation over Senegal for 2005. Results showed a specific pattern of rainfall throughout the Dakar region during August, 2005, and the associated rainfall anomaly coincided with an exacerbation of the cholera epidemic. Comparison of rainfall and epidemiological patterns revealed that the temporal dynamics of precipitation, which was abrupt and heavy, was presumably the determining factor. Analysis of the SST gradient showed that the Atlantic Ocean SST variability in 2005 differed from that of 2002 to 2004, a result of a prominent Atlantic meridional mode. The influence of this intense precipitation on cholera transmission over a densely populated and crowded region was detectable for both Dakar and Thiès, Senegal. Thus, high resolution rainfall forecasts at subseasonal time scales should provide a way forward for an early warning system in Africa for cholera and, thereby, trigger epidemic preparedness. Clearly, attention must be paid to both natural and human induced environmental factors to devise appropriate action to prevent cholera and other waterborne disease epidemics in the region. PMID:22952995

  11. Predictive modeling of cholera using GRACE and TRMM satellite data

    Science.gov (United States)

    Jutla, A.; Akanda, A. S. S.; Colwell, R. R.

    2015-12-01

    Cholera outbreaks can be classified in three forms- epidemic (sudden or seasonal outbreaks), endemic (recurrence and persistence of the disease for several consecutive years) and mixed-mode endemic (combination of certain epidemic and endemic conditions) with significant spatial and temporal heterogeneity. Endemic cholera is related to floods and droughts in regions where water and sanitation infrastructure are inadequate or insufficient. With more than a decade of terrestrial water storage (TWS) data obtained from Gravity Recovery and Climate Experiment (GRACE), understanding dynamics of river discharge is now feasible. We explored lead-lag relationships between TWS in the Ganges-Brahmaputra-Meghna (GBM) basin and endemic cholera in Bangladesh. Since bimodal seasonal peaks in cholera in Bangladesh occur during the spring and autumn season, two separate models, between TWS and disease time series (2002 to 2010) were developed. TWS, hence water availability, showed an asymmetrical, strong association with spring (τ=-0.53; p<0.001) and autumn (τ=0.45; p<0.001) cholera prevalence up to five to six months in advance. One unit (cm of water) decrease in water availability in the basin increased odds of above normal cholera by 24% [confidence interval (CI) 20-31%; p<0.05] in the spring season, while an increase in regional water by one unit, through floods, increased odds of above average cholera in the autumn by 29% [CI:22-33%; p<0.05]. Epidemic cholera is related with warm temperatures and heavy rainfall. Using TRMM data for several locations in Asia and Africa, probability of cholera increases 18% [CI:15-23%; p<0.05] after heavy precipitation resulted in a societal conditions where access to safe water and sanitation was disrupted. Results from mechanistic modeling framework using systems approach that include satellite based hydroclimatic information with tradition disease transmission models will also be presented.

  12. Study on the monitoring methods of Vibrio cholerae in aquatic products%水产品中霍乱弧菌的监测方法研究

    Institute of Scientific and Technical Information of China (English)

    谢朝梅; 胡世雄; 邓志红; 湛志飞; 华伟湘; 熊伯华; 谢燕湘; 邓海斌; 卜昕琳

    2012-01-01

    Objective:To study the monitoring methods of Vibrio cholerae in aquatic products, find the aquatic products contaminated by Vibrio cholerae timely, and take effective measures to prevent and control cholera outbreak. Methods:The real - time fluorescence PCR, the colloidal gold method, the isolation and culture method were used for Vibrio cholerae detection in 180 aquatic product samples respectively. Results; With the three methods, 44 nucleic acid positive specimens were detected, in which 18 specimens were cholera enterotoxin positive, and 8 Vibrio cholerae strains were isolated. Conclusion; During cholera surveillance of aquatic products, nucleic acid positive samples were screened first by real - time PCR, and then cholera enterotoxin detection was done to find the polluted aquatic products timely. Colloidal gold method was employed to isolate, culture and type the nucleic acid positive samples for further confirmation and analysis of cholera spread.%目的:研究水产品中霍乱弧菌的监测方法,及时发现被霍乱弧菌污染的水产品,采取有效防控措施,防范霍乱疫情的发生.方法:用实时荧光PCR法、胶体金法、分离培养法三种方法分别对180份水产品进行霍乱弧菌检测,阳性样品再进行霍乱肠毒素检测.结果:三种方法检出了44份核酸阳性标本,18份霍乱肠毒素阳性标本,分离出8株霍乱弧菌.结论:在进行水产品霍乱监测时,可以先用实时荧光PCR进行初筛,筛出核酸阳性标本,进行霍乱肠毒素检测,及时发现被霍乱产毒株污染的水产品,再对筛出的核酸阳性标本结合胶体金法进行分离培养和分型鉴定,进一步完成霍乱疫情的确证和分析.

  13. Concurrent outbreaks of cholera and peripheral neuropathy associated with high mortality among persons internally displaced by a volcanic eruption.

    Directory of Open Access Journals (Sweden)

    Alexander Rosewell

    Full Text Available BACKGROUND: In October 2004, Manam Island volcano in Papua New Guinea erupted, causing over 10 000 villagers to flee to internally displaced person (IDP camps, including 550 from Dugulaba village. Following violence over land access in March 2010, the IDPs fled the camps, and four months later concurrent outbreaks of acute watery diarrhea and unusual neurological complaints were reported in this population. MATERIALS AND METHODS: A retrospective case-control study was conducted to identify the risk factors for peripheral neuropathy. Rectal swabs were collected from cases of acute watery diarrhea. Hair and serum metals and metalloids were analyzed by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS. RESULTS: There were 17 deaths among the 550 village inhabitants during the outbreak period at a crude mortality rate 21-fold that of a humanitarian crisis. Vibrio cholerae O1 El Tor Ogawa was confirmed among the population. Access to community-level rehydration was crucial to mortality. Peripheral neuropathy was diagnosed among cases with neurological symptoms. A balanced diet was significantly protective against neuropathy. A dose-response relationship was seen between peripheral neuropathy and a decreasing number of micronutrient- rich foods in the diet. Deficiencies in copper, iron, selenium and zinc were identified among the cases of peripheral neuropathy. CONCLUSIONS: Cholera likely caused the mostly preventable excess mortality. Peripheral neuropathy was not caused by cholera, but cholera may worsen existing nutritional deficiencies. The peripheral neuropathy was likely caused by complex micronutrient deficiencies linked to non-diversified diets that potentially increased the vulnerability of this population, however a new zinc-associated neuropathy could not be ruled out. Reoccurrence can be prevented by addressing the root cause of displacement and ensuring access to arable land and timely resettlement.

  14. New developments in the understanding of cholera.

    Science.gov (United States)

    Butler, T

    2001-08-01

    Recent advances in prevention and treatment of cholera have occurred in the areas of vaccine testing, modifications of oral-rehydration solutions (ORS), and antimicrobial treatment. Oral vaccines consisting of killed whole bacterial cells (WC) with and without the B-subunit of cholera toxin (BS) were shown to be effective in large trials in Bangladesh, Peru, and Vietnam. However, the trials did not resolve whether two or three doses of vaccine are required and whether BS adds significantly to the immune protection of WC. Live, attenuated bacterial vaccines that are immunogenic and have been shown protective in human volunteer studies are candidates for future field trials. Rehydration of patients is a life- saving effort. The best ORS contains rice powder in place of glucose, and solutions with reduced osmolarity (245 mOsm/L, sodium 75 mEq/L) are as effective as standard ORS. Ciprofloxacin in a single dose is effective in adults, and erythromycin or ampicillin in multiple doses is effective in children. PMID:11470000

  15. Single-molecule tracking in live Vibrio cholerae reveals that ToxR recruits the membrane-bound virulence regulator TcpP to the toxT promoter.

    Science.gov (United States)

    Haas, Beth L; Matson, Jyl S; DiRita, Victor J; Biteen, Julie S

    2015-04-01

    Vibrio cholerae causes the human disease cholera by producing a potent toxin. The V. cholerae virulence pathway involves an unusual transcription step: the bitopic inner-membrane proteins TcpP and ToxR activate toxT transcription. As ToxT is the primary direct transcription activator in V. cholerae pathogenicity, its regulation by membrane-localized activators is key in the disease process. However, the molecular mechanisms by which membrane-localized activators engage the transcription process have yet to be uncovered in live cells. Here we report the use of super-resolution microscopy, single-molecule tracking, and gene knockouts to examine the dynamics of individual TcpP proteins in live V. cholerae cells with < 40 nm spatial resolution on a 50 ms timescale. Single-molecule trajectory analysis reveals that TcpP diffusion is heterogeneous and can be described by three populations of TcpP motion: one fast, one slow, and one immobile. By comparing TcpP diffusion in wild-type V. cholerae to that in mutant strains lacking either toxR or the toxT promoter, we determine that TcpP mobility is greater in the presence of its interaction partners than in their absence. Our findings support a mechanism in which ToxR recruits TcpP to the toxT promoter for transcription activation.

  16. Exoproteome and secretome derived broad spectrum novel drug and vaccine candidates in Vibrio cholerae targeted by Piper betel derived compounds.

    Directory of Open Access Journals (Sweden)

    Debmalya Barh

    Full Text Available Vibrio cholerae is the causal organism of the cholera epidemic, which is mostly prevalent in developing and underdeveloped countries. However, incidences of cholera in developed countries are also alarming. Because of the emergence of new drug-resistant strains, even though several generic drugs and vaccines have been developed over time, Vibrio infections remain a global health problem that appeals for the development of novel drugs and vaccines against the pathogen. Here, applying comparative proteomic and reverse vaccinology approaches to the exoproteome and secretome of the pathogen, we have identified three candidate targets (ompU, uppP and yajC for most of the pathogenic Vibrio strains. Two targets (uppP and yajC are novel to Vibrio, and two targets (uppP and ompU can be used to develop both drugs and vaccines (dual targets against broad spectrum Vibrio serotypes. Using our novel computational approach, we have identified three peptide vaccine candidates that have high potential to induce both B- and T-cell-mediated immune responses from our identified two dual targets. These two targets were modeled and subjected to virtual screening against natural compounds derived from Piper betel. Seven compounds were identified first time from Piper betel to be highly effective to render the function of these targets to identify them as emerging potential drugs against Vibrio. Our preliminary validation suggests that these identified peptide vaccines and betel compounds are highly effective against Vibrio cholerae. Currently we are exhaustively validating these targets, candidate peptide vaccines, and betel derived lead compounds against a number of Vibrio species.

  17. Cholera Toxin B: One Subunit with Many Pharmaceutical Applications

    Directory of Open Access Journals (Sweden)

    Keegan J. Baldauf

    2015-03-01

    Full Text Available Cholera, a waterborne acute diarrheal disease caused by Vibrio cholerae, remains prevalent in underdeveloped countries and is a serious health threat to those living in unsanitary conditions. The major virulence factor is cholera toxin (CT, which consists of two subunits: the A subunit (CTA and the B subunit (CTB. CTB is a 55 kD homopentameric, non-toxic protein binding to the GM1 ganglioside on mammalian cells with high affinity. Currently, recombinantly produced CTB is used as a component of an internationally licensed oral cholera vaccine, as the protein induces potent humoral immunity that can neutralize CT in the gut. Additionally, recent studies have revealed that CTB administration leads to the induction of anti-inflammatory mechanisms in vivo. This review will cover the potential of CTB as an immunomodulatory and anti-inflammatory agent. We will also summarize various recombinant expression systems available for recombinant CTB bioproduction.

  18. Pandemic serotypes of Vibrio cholerae isolated from ships' ballast tanks and coastal waters: assessment of antibiotic resistance and virulence genes (tcpA and ctxA).

    Science.gov (United States)

    Dobbs, Fred C; Goodrich, Amanda L; Thomson, Frank K; Hynes, Wayne

    2013-05-01

    There is concern that ships' ballasting operations may disseminate Vibrio cholerae to ports throughout the world. Given evidence that the bacterium is indeed transported by ships, we isolated pandemic serotypes O1 and O139 from ballast tanks and characterized them with respect to antibiotic resistance and virulence genes ctxA and tcpA. We carried out concurrent studies with V. cholerae isolated from coastal waters. Of 284 isolates, 30 were serotype O1 and 59 were serotype O139. These serotypes were overrepresented in ballast tanks relative to the coastal waters sampled. All locations, whether coastal waters or ballast tanks, yielded samples from which serotype O1, O139, or both were isolated. There were three groups among the 62 isolates for which antibiotic characterization was conclusive: those exhibiting β-lactamase activity and resistance to at least one of the 12 antibiotics tested; those negative for β-lactamase but having antibiotic resistance; those negative for β-lactamase and registering no antibiotic resistance. When present, antibiotic resistance in nearly all cases was to ampicillin; resistance to multiple antibiotics was uncommon. PCR assays revealed that none of the isolates contained the ctxA gene and only two isolates, one O139 and one O1, contained the tcpA gene; both isolates originated from ballast water. These results support the bacteriological regulations proposed by the International Maritime Association for discharged ballast water.

  19. The Role of Cyanobacteria Blooms in Cholera Epidemic in Bangladesh

    Science.gov (United States)

    Sagir Ahmed, Md.; Raknuzzaman, Md.; Akther, Hafeza; Ahmed, Sumaiya

    A study was conducted on association of Vibrio cholerae with plankton specially emphasis on cyanobacteria in relation to some physico-chemical parameters in the River Buriganga, Dhaka, from January to December 2002. Monthly abundance of phytoplankton and zooplankton varied from 457 to 14166 and from 169 to 1055 individual L-1, respectively. Monthly average of faecal coliform in water, zooplankton and phytoplankton samples were 3.99x109, 4.54x103 and 4.28x102 (CFU L-1), respectively. During epidemics, toxigenic V. cholerae 01 and 0139 were isolated from the patients as well as from the surface water. V. cholerae 01 and 0139 were also isolated from plankton samples. More over, it was observed that ctx (cholera toxic) positive in water and phytoplankton samples of the river. A bloom of Oscillatoria sp. (1.6x104 individual L-1) occurred in the upper reaches of the River Buriganga in May 2002. Methanol-water extract of bloom sample was analyzed by high performance liquid chromatography with UV detection and Mass Spectrum (MS) detected microcystin-RR. Cyanobacteria are abundant in the aquatic environment of Bangladesh and it was established that V. cholerae maintain a symbiotic relationship with these algae particularly mucilaginous cyanobacteria. During epidemics, patients symptoms included diarrhea, vomiting and hemorrhagic enteritis and in severe cases hemorrhagic diarrhea. So, question has arisen that which is responsible, microcystins or cholera for death of cholera/diarrhea patients in Bangladesh. Future research should be directed to isolate microcystins and cholera toxins from the epidemic areas to clarify the fact.

  20. Cholera in Ecuador: Current relevance of past lessons learnt

    OpenAIRE

    S S Malavade; A Narvaez; A. Mitra; Ochoa, T; Naik, E.; Sharma, M.; S Galwankar; M D Breglia; Izurieta, R

    2011-01-01

    This report analyses the trends in the cholera epidemic that hit Ecuador in 1991. The study is based on personal experiences and analysis of epidemiological databases from the Ministry of Public Health of Ecuador. The number of cases and initial attack rates in an immunologically naive population are described by province. An analysis of the Andean and coastal cholera patterns of transmission are described along with its associated risk factors. The logistical, environmental, and socio-cultur...

  1. Community health facility preparedness for a cholera surge in Haiti.

    Science.gov (United States)

    Mobula, Linda Meta; Jacquet, Gabrielle A; Weinhauer, Kristin; Alcidas, Gladys; Thomas, Hans-Muller; Burnham, Gilbert

    2013-01-01

    With increasing population displacement and worsening water insecurity after the 2010 earthquake, Haiti experienced a large cholera outbreak. Our goal was to evaluate the strengths and weaknesses of seven community health facilities' ability to respond to a surge in cholera cases. Since 2010, Catholic Relief Services (CRS) with a number of public and private donors has been working with seven health facilities in an effort to reduce morbidity and mortality from cholera infection. In November 2012, CRS through the Centers for Disease Control and Prevention (CDC)'s support, asked the Johns Hopkins Center for Refugee and Disaster Response to conduct a cholera surge simulation tabletop exercise at these health facilities to improve each facility's response in the event of a cholera surge. Using simulation development guidelines from the Pan American Health Organization and others, a simulation scenario script was produced that included situations of differing severity, supply chain, as well as a surge of patients. A total of 119 hospital staff from seven sites participated in the simulation exercise including community health workers, clinicians, managers, pharmacists, cleaners, and security guards. Clinics that had challenges during the simulated clinical care of patients were those that did not appropriately treat all cholera patients according to protocol, particularly those that were vulnerable, those that would need additional staff to properly treat patients during a surge of cholera, and those that required a better inventory of supplies. Simulation-based activities have the potential to identify healthcare delivery system vulnerabilities that are amenable to intervention prior to a cholera surge. PMID:24481887

  2. Warming Oceans, Phytoplankton, and River Discharge: Implications for Cholera Outbreaks

    OpenAIRE

    Jutla, Antarpreet S; Akanda, Ali S.; Griffiths, Jeffrey K; Colwell, Rita; Islam, Shafiqul

    2011-01-01

    Phytoplankton abundance is inversely related to sea surface temperature (SST). However, a positive relationship is observed between SST and phytoplankton abundance in coastal waters of Bay of Bengal. This has led to an assertion that in a warming climate, rise in SST may increase phytoplankton blooms and, therefore, cholera outbreaks. Here, we explain why a positive SST-phytoplankton relationship exists in the Bay of Bengal and the implications of such a relationship on cholera dynamics. We f...

  3. Analysis on monitoring data of vibrio cholerae in Yulin during 2003 to 2010%2003-2010年玉林市霍乱弧菌监测结果分析

    Institute of Scientific and Technical Information of China (English)

    李劲锋; 李文; 权怡; 梁炯明; 王鸣柳; 刘义威; 罗铭; 胡昱

    2011-01-01

    Objective To analyze the monitoring data of vibrio cholerae from 2003 to 2010 in Yulin, and provide scientific basis for prevention and control the disease. Method According to strategy of surveillance of cholera in China and the fifth edition of manuals of vibrio cholera, detection, isolation, and identification the samples of external environment water, fishery products , food and diarrhea patients and focus groups. Positive strains were carried out drug resistance analysis, and detected Ctx and Zot virulence genes by PCR nucleic acid. Results 23 391 samples were collected during 2003 to 2010 and vibrio cholera were found in 27 samples, positive rate was 0. 12%. Among them, the positive rate of water was 0. 23% , positive samples account for 25.93%. Fishery products were 0. 40% . account for 74. 07%㏑ and the positive rate of frogs were highest than others ( 4. 18% ) . Moreover, vibrio cholera did not detected from food, diarrhea patients and focus groups. Serotypes showed that there were 16 inaba samples ( account for 59. 26% ) , 3 ogawa samples ( 11. 11 % ) . 4 hikojima samples (14. 81% ) and 4 O139 ( 14. 81% ) . Virulence gene detection showed that 75% O139 carried Ctx and Zot genes, and accounted for ( 3/4) , O1 group did not carry Ctx, but 30. 43% (7/23) carried Zot. 27 strains were all sensitive to norfloxacin, ciprofloxacin, amikacin, cefalotin, ceftazidime, and drug resistance to streptomycin, tetracycline,sulfa. Conclusions V. cholera existed in external environmental in Yulin city, we should enhance monitoring, study toxic gene charactenstics, leam drug resistance changes, to enhance scientific prediction and pertinence of prevention and control work.%目的 分析玉林市2003-2010霍乱监测的情况,为今后预防控制策略提供科学依据.方法 按及(第5版)的规定,对外环境水体、海(水)产品、食品及腹泻病人和重点人群进行采样监测、分离、鉴定,阳性株进行耐药性分析,PCR核

  4. The repertoire of glycosphingolipids recognized by Vibrio cholerae.

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    John Benktander

    Full Text Available The binding of cholera toxin to the ganglioside GM1 as the initial step in the process leading to diarrhea is nowadays textbook knowledge. In contrast, the knowledge about the mechanisms for attachment of Vibrio cholerae bacterial cells to the intestinal epithelium is limited. In order to clarify this issue, a large number of glycosphingolipid mixtures were screened for binding of El Tor V. cholerae. Several specific interactions with minor complex non-acid glycosphingolipids were thereby detected. After isolation of binding-active glycosphingolipids, characterization by mass spectrometry and proton NMR, and comparative binding studies, three distinct glycosphingolipid binding patterns were defined. Firstly, V. cholerae bound to complex lacto/neolacto glycosphingolipids with the GlcNAcβ3Galβ4GlcNAc sequence as the minimal binding epitope. Secondly, glycosphingolipids with a terminal Galα3Galα3Gal moiety were recognized, and the third specificity was the binding to lactosylceramide and related compounds. V. cholerae binding to lacto/neolacto glycosphingolipids, and to the other classes of binding-active compounds, remained after deletion of the chitin binding protein GbpA. Thus, the binding of V. cholerae to chitin and to lacto/neolacto containing glycosphingolipids represents two separate binding specificities.

  5. Surveillance on Vibrio Cholera isolated from water and aquatic products in Guangxi,2006 - 2009%2006年-2009年广西水体及海水产品霍乱弧菌监测分析

    Institute of Scientific and Technical Information of China (English)

    权怡; 方锦嵩; 周凌云; 林玫; 王鸣柳; 秦卫文

    2011-01-01

    Objective:To early detect the source and control the outbreaks via surveillance on Vibrio Cholera in aquatic products. Methods: Aquatic products were collected and cultured for Vibrio Cholera. The isolates from diverse sources were identified by serology, phage typing, antibiotic susceptibility tests and molecular biology. Results: A total of 75 strains of Vibrio Cholera were isolated between 2006 and 2009, of which, 17 were identified Vibrio Cholera serotype Ogawa, 48 Vibrio Cholera serotype Inaba and 5 Vibrio Cholera serogroup O139. Five strains of Vibrio Cholera serogtype Hikojima was detected for the first time. Cholera enterotoxin (ctxA)was detected in 3 strains of Vibrio Cholera O139 out of 75 strains isolated, while the remaining were non -toxigenic strains. The results of antimicrobiai susceptibility tests showed that the strains were susceptible to norfloxacin, aminkacin, ciprofloxacin, ceftazidime and cefalothin, and were resistant to gentamycin, SMZ - Co, nitrofurantoin, tetracycline, ampicillin, doxycycline, chloramphenicol, streptomycin and sulfanilamide. Conclusion: Vibrio Cholera was detected in the environment in Guangxi. The vibrio isolated were mainly non - toxigenic strains. Resistance was found in different serotypes of Vibrio Cholera.No outbreak of cholera was observed in this area.%目的:通过水体及海水产品的霍乱弧菌监测,及时发现疫情传播来源,以有效控制霍乱疫情的发生.方法:采集各类海水产品分离培养霍乱弧菌,采用血清学、嗜菌体生物分型、药物敏感试验和分子生物学方法对不同来源的菌株进行鉴定.结果:2006年-2009年共分离出75株霍乱弧菌,其中小川型霍乱弧菌17株,稻叶型霍乱弧菌48株,O139群霍乱弧菌5株,首次分离出彦岛型霍乱弧菌5株.75株霍乱弧菌中仅3株O139群霍乱弧菌携带霍乱肠毒素(ctxA),其余菌株均为非流行株或非产毒株.药敏结果显示,除对氟哌酸、阿米卡星、环丙沙星、头孢

  6. Elevation and cholera: an epidemiological spatial analysis of the cholera epidemic in Harare, Zimbabwe, 2008-2009

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    Luque Fernandez Miguel A

    2012-06-01

    Full Text Available Abstract Background In highly populated African urban areas where access to clean water is a challenge, water source contamination is one of the most cited risk factors in a cholera epidemic. During the rainy season, where there is either no sewage disposal or working sewer system, runoff of rains follows the slopes and gets into the lower parts of towns where shallow wells could easily become contaminated by excretes. In cholera endemic areas, spatial information about topographical elevation could help to guide preventive interventions. This study aims to analyze the association between topographic elevation and the distribution of cholera cases in Harare during the cholera epidemic in 2008 and 2009. Methods We developed an ecological study using secondary data. First, we described attack rates by suburb and then calculated rate ratios using whole Harare as reference. We illustrated the average elevation and cholera cases by suburbs using geographical information. Finally, we estimated a generalized linear mixed model (under the assumption of a Poisson distribution with an Empirical Bayesian approach to model the relation between the risk of cholera and the elevation in meters in Harare. We used a random intercept to allow for spatial correlation of neighboring suburbs. Results This study identifies a spatial pattern of the distribution of cholera cases in the Harare epidemic, characterized by a lower cholera risk in the highest elevation suburbs of Harare. The generalized linear mixed model showed that for each 100 meters of increase in the topographical elevation, the cholera risk was 30% lower with a rate ratio of 0.70 (95% confidence interval=0.66-0.76. Sensitivity analysis confirmed the risk reduction with an overall estimate of the rate ratio between 20% and 40%. Conclusion This study highlights the importance of considering topographical elevation as a geographical and environmental risk factor in order to plan cholera preventive

  7. Hybrid microarray based on double biomolecular markers of DNA and carbohydrate for simultaneous genotypic and phenotypic detection of cholera toxin-producing Vibrio cholerae.

    Science.gov (United States)

    Shin, Hwa Hui; Seo, Jeong Hyun; Kim, Chang Sup; Hwang, Byeong Hee; Cha, Hyung Joon

    2016-05-15

    Life-threatening diarrheal cholera is usually caused by water or food contaminated with cholera toxin-producing Vibrio cholerae. For the prevention and surveillance of cholera, it is crucial to rapidly and precisely detect and identify the etiological causes, such as V. cholerae and/or its toxin. In the present work, we propose the use of a hybrid double biomolecular marker (DBM) microarray containing 16S rRNA-based DNA capture probe to genotypically identify V. cholerae and GM1 pentasaccharide capture probe to phenotypically detect cholera toxin. We employed a simple sample preparation method to directly obtain genomic DNA and secreted cholera toxin as target materials from bacterial cells. By utilizing the constructed DBM microarray and prepared samples, V. cholerae and cholera toxin were detected successfully, selectively, and simultaneously; the DBM microarray was able to analyze the pathogenicity of the identified V. cholerae regardless of whether the bacteria produces toxin. Therefore, our proposed DBM microarray is a new effective platform for identifying bacteria and analyzing bacterial pathogenicity simultaneously. PMID:26735874

  8. Cholera at the Crossroads: The Association Between Endemic Cholera and National Access to Improved Water Sources and Sanitation

    OpenAIRE

    Nygren, Benjamin L.; Blackstock, Anna J.; Eric D Mintz

    2014-01-01

    We evaluated World Health Organization (WHO) national water and sanitation coverage levels and the infant mortality rate as predictors of endemic cholera in the 5-year period following water and sanitation coverage estimates using logistic regression, receiver operator characteristic curves, and different definitions of endemicity. Each was a significant predictors of endemic cholera at P < 0.001. Using a value of 250 for annual cases reported in 3 of 5 years, a national water access level of...

  9. Characterization of tryptophanase from Vibrio cholerae.

    Science.gov (United States)

    Nuidate, Taiyeebah; Tansila, Natta; Chomchuen, Piraporn; Phattaranit, Phattiphong; Eangchuan, Supachok; Vuddhakul, Varaporn

    2015-01-01

    Tryptophanase (Trpase) is a pyridoxal phosphate (PLP)-dependent enzyme responsible for the production of indole, an important intra- and interspecies signaling molecule in bacteria. In this study, the tnaA gene of Vibrio cholerae coding for VcTrpase was cloned into the pET-20b(+) vector and expressed in Escherichia coli BL21(DE3) tn5:tnaA. Using Ni(2+)-nitrilotriacetic acid (NTA) chromatography, VcTrpase was purified, and it possessed a molecular mass of ∼49 kDa with specific absorption peaks at 330 and 435 nm and a specific activity of 3 U/mg protein. The VcTrpase had an 80 % homology to the Trpase of Haemophilus influenzae and E. coli, but only around 50 % identity to the Trpase of Proteus vulgaris and Porphyromonas gingivalis. The optimum conditions for the enzyme were at pH 9.0 and 45 °C. Recombinant VcTrpase exhibited analogous kinetic reactivity to the EcTrpase with K m and k cat values of 0.612 × 10(-3) M and 5.252 s(-1), respectively. The enzyme catalyzed S-methyl-L-cysteine and S-benzyl-L-cysteine degradation, but not L-phenylalanine and L-serine. Using a site-directed mutagenesis technique, eight residues (Thr52, Tyr74, Arg103, Asp137, Arg230, Lys269, Lys270, and His463) were conserved for maintaining enzyme catalysis. All amino acid substitutions at these sites either eliminated or remarkably diminished Trpase activity. These sites are thus potential targets for the design of drugs to control the V. cholerae Trpase and to further investigate its functions.

  10. Bile salt-induced intermolecular disulfide bond formation activates Vibrio cholerae virulence.

    Science.gov (United States)

    Yang, Menghua; Liu, Zhi; Hughes, Chambers; Stern, Andrew M; Wang, Hui; Zhong, Zengtao; Kan, Biao; Fenical, William; Zhu, Jun

    2013-02-01

    To be successful pathogens, bacteria must often restrict the expression of virulence genes to host environments. This requires a physical or chemical marker of the host environment as well as a cognate bacterial system for sensing the presence of a host to appropriately time the activation of virulence. However, there have been remarkably few such signal-sensor pairs identified, and the molecular mechanisms for host-sensing are virtually unknown. By directly applying a reporter strain of Vibrio cholerae, the causative agent of cholera, to a thin layer chromatography (TLC) plate containing mouse intestinal extracts, we found two host signals that activate virulence gene transcription. One of these was revealed to be the bile salt taurocholate. We then show that a set of bile salts cause dimerization of the transmembrane transcription factor TcpP by inducing intermolecular disulfide bonds between cysteine (C)-207 residues in its periplasmic domain. Various genetic and biochemical analyses led us to propose a model in which the other cysteine in the periplasmic domain, C218, forms an inhibitory intramolecular disulfide bond with C207 that must be isomerized to form the active C207-C207 intermolecular bond. We then found bile salt-dependent effects of these cysteine mutations on survival in vivo, correlating to our in vitro model. Our results are a demonstration of a mechanism for direct activation of the V. cholerae virulence cascade by a host signal molecule. They further provide a paradigm for recognition of the host environment in pathogenic bacteria through periplasmic cysteine oxidation.

  11. Vibriocidal antibody responses to a bivalent killed whole-cell oral cholera vaccine in a phase III trial in Kolkata, India.

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    Suman Kanungo

    Full Text Available BACKGROUND: During the development of a vaccine, identification of the correlates of protection is of paramount importance for establishing an objective criterion for the protective performance of the vaccine. However, the ascertainment of correlates of immunity conferred by any vaccine is a difficult task. METHODS: While conducting a phase three double-blind, cluster-randomized, placebo-controlled trial of a bivalent killed whole-cell oral cholera vaccine in Kolkata, we evaluated the immunogenicity of the vaccine in a subset of participants. Randomly chosen participants (recipients of vaccine or placebo were invited to provide blood samples at baseline, 14 days after the second dose and one year after the first dose. At these time points, serum geometric mean titers (GMT of vibriocidal antibodies and seroconversion rates for vaccine and placebo arms were calculated and compared across the age strata (1 to 5 years, 5 to 15 years and more than 15 years as well as for all age groups. RESULTS: Out of 137 subjects included in analysis, 69 were vaccinees and 68 received placebo. There were 5•7 and 5•8 geometric mean fold (GMF rises in titers to Vibrio cholerae Inaba and Ogawa, respectively at 14 days after the second dose, with 57% and 61% of vaccinees showing a four-fold or greater titer rise, respectively. After one year, the titers to Inaba and Ogawa remained 1•7 and 2•8 fold higher, respectively, compared to baseline. Serum vibriocidal antibody response to V. cholerae O139 was much lower than that to Inaba or Ogawa. No significant differences in the GMF-rises were observed among the age groups. CONCLUSIONS: The reformulated oral cholera vaccine induced a statistically significant anti-O1 Inaba and O1 Ogawa vibriocidal antibody response 14 days after vaccination, which although declined after one year remained significantly higher than baseline. Despite this decline, the vaccine remained protective five years after vaccination.

  12. Vibrio cholerae O139: Surgimiento, evolución y estructura genética de CTXO

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    Talena Ledón

    2007-01-01

    Full Text Available El surgimiento de Vibrio cholerae O139 Bengal entre 1992 y 1993 estuvo asociado con grandes brotes epidémicos en la India y Bangladesh. En un inicio, estas cepas lograron desplazar a las de Vibrio cholerae O1 circulantes, responsables hasta ese momento de los casos de cólera reportados. No obstante, los vibrios O1 emergieron nuevamente en 1994 y se iniciaron ciclos de mayor y menor incidencia de Vibrio cholerae O139. Esta intermitencia epidemiológica se ha relacionado con cambios genéticos y fenotípicos en las cepas de Vibrio cholerae O139. El análisis genético molecular del polimorfismo de los fragmentos de restricción ha sido una herramienta muy útil para estudiar los cambios mencionados anteriormente. Estos estudios apuntan a la existencia de una diversidad clonal entre las cepas O139, lo cual se revela en las diversas estructuras en que se presenta el fago CTX Fi en el genoma de estas cepas, las cuales incluyen cambios en el número de copias, en la localización en el genoma y en la propia estructura de CTX Fi. En este trabajo se hace una revisión de la información disponible sobre este tema y se reporta la estructura genética de CTX Fi en cepas de reciente aislamiento de la India, en las cuales existen copias de CTX Fi intactas y carentes de toxina colérica coexistiendo en la misma cepa. Las copias de CTX Fi ctx-negativas pertenecen a los tipos Calcuta y El Tor y pudieran considerarse precursores de ambas clases de CTX Fi, respectivamente. Un análisis comparativo entre parte de la secuencia de ellos y lo reportado por otros autores, indicó diferencias en sus orígenes. Las cepas analizadas poseen además, copias diferentes de CTX Fi con el mismo grupo de inmunidad. En el artículo se discute la implicación de este hecho sobre el concepto de heteroinmunidad y en el diseño de vacunas.

  13. Detection and characterization of integrative and conjugative elements (ICEs)-positive Vibrio cholerae isolates from aquacultured shrimp and the environment in Shanghai, China.

    Science.gov (United States)

    He, Yu; Tang, Yuyi; Sun, Fengjiao; Chen, Lanming

    2015-12-30

    Increasing industrialization and use of antimicrobial agents in aquaculture production, have led to heavy metals and multidrug resistant (MDR) pathogens becoming serious problems. These resistances are conferred in two ways: intrinsic and transfer via conjugation, or transformation by the major transmission mediators. Integrative and conjugative elements (ICEs) are one of the major mediators; however, few studies on ICEs of environmental origin have been reported in Asia. Herein, we determined the prevalence, antimicrobial susceptibility, heavy metal resistance and genotypes of 126 strains of Vibrio cholerae isolated from aquatic products and the environment in Shanghai, China. 92.3% of isolates were ICEs-positive from aquaculture water and 89.3% of isolates from shrimp showed MDR. Tracing the V. cholerae genotypes, showed no significant relevance of genotype among the antimicrobial resistance strains bearing the ICEs or not. Thus, in aquaculture, ICEs are not the major transmission mediators of resistance to antibiotics or heavy metals.

  14. RpoS plays a central role in the SOS induction by sub-lethal aminoglycoside concentrations in Vibrio cholerae.

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    Zeynep Baharoglu

    Full Text Available Bacteria encounter sub-inhibitory concentrations of antibiotics in various niches, where these low doses play a key role for antibiotic resistance selection. However, the physiological effects of these sub-lethal concentrations and their observed connection to the cellular mechanisms generating genetic diversification are still poorly understood. It is known that, unlike for the model bacterium Escherichia coli, sub-minimal inhibitory concentrations (sub-MIC of aminoglycosides (AGs induce the SOS response in Vibrio cholerae. SOS is induced upon DNA damage, and since AGs do not directly target DNA, we addressed two issues in this study: how sub-MIC AGs induce SOS in V. cholerae and why they do not do so in E. coli. We found that when bacteria are grown with tobramycin at a concentration 100-fold below the MIC, intracellular reactive oxygen species strongly increase in V. cholerae but not in E. coli. Using flow cytometry and gfp fusions with the SOS regulated promoter of intIA, we followed AG-dependent SOS induction. Testing the different mutation repair pathways, we found that over-expression of the base excision repair (BER pathway protein MutY relieved this SOS induction in V. cholerae, suggesting a role for oxidized guanine in AG-mediated indirect DNA damage. As a corollary, we established that a BER pathway deficient E. coli strain induces SOS in response to sub-MIC AGs. We finally demonstrate that the RpoS general stress regulator prevents oxidative stress-mediated DNA damage formation in E. coli. We further show that AG-mediated SOS induction is conserved among the distantly related Gram negative pathogens Klebsiella pneumoniae and Photorhabdus luminescens, suggesting that E. coli is more of an exception than a paradigm for the physiological response to antibiotics sub-MIC.

  15. ANTAGONISM AGAINST VIBRIO CHOLERAE BY BACTERIAL DIFFUSIBLE COMPOUND IN THE FECAL MICROBIOTA OF RODENTS

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    Simone Helena da Silva

    1998-09-01

    Full Text Available In an ex vivo agar plate assay, we monitored the appearance of an inhibitory halo against Vibrio cholerae from the feces of Wistar and Fischer rats aged 10 to 42 days. The frequency of Wistar rats showing halo increased from 0% (10 days to a maximum of 80.0% (29 days and then decreased to 53.3% (42 days. A similar pattern was obtained with Fischer rats but with a lower intensity (maximum frequency of 50.0% by day 36. In a separate experiment, when Wistar rats were fed a low-protein diet for 7 days, the inhibitory halo decreased drastically. Three apparently different colony morphologies were isolated from the dominant fecal microbiota: a facultative anaerobe (FAN and two strict anaerobes (SAN. The ex vivo inhibitory test showed a halo around the feces of germfree mice monoassociated with the FAN bacterium or one of the SAN bacterium but not of the germfree ones. After oral challenge of all groups with V. cholerae, a permissive and a drastic barrier effects were observed in mice with FAN and SAN associated bacteria, respectively. The FAN and one SAN bacteria used in the in vivo challenges were identified as Escherichia coli and Streptococcus intermedius, respectively. The potent antagonism developed by the rat intestinal microbiota against V. cholerae seems to be due, in part, to diffusible compounds and this phenomenon depends apparently on age, strain and nutrition of the animals. These preliminary results also suggest that this effect was due to more than one bacterial component at any given moment.O aparecimento de halo de inibição contra o Vibrio cholerae a partir das fezes de ratos Wistar e Fischer nas idades de 10 a 42 dias foi observado usando um teste ex vivo em placa. A frequência de ratos Wistar apresentando halo aumentou de 0% (10 dias até um máximo de 80,0% (29 dias antes de decair para 53,3% (42 dias. Um perfil similar foi obtido com os ratos Fischer mas com valores inferiores (frequência máxima de 50,0% no dia 36. Num

  16. The Transmission and Antibiotic Resistance Variation in a Multiple Drug Resistance Clade of Vibrio cholerae Circulating in Multiple Countries in Asia.

    Science.gov (United States)

    Pang, Bo; Du, Pengcheng; Zhou, Zhemin; Diao, Baowei; Cui, Zhigang; Zhou, Haijian; Kan, Biao

    2016-01-01

    Vibrio cholerae has caused massive outbreaks and even trans-continental epidemics. In 2008 and 2010, at least 3 remarkable cholera outbreaks occurred in Hainan, Anhui and Jiangsu provinces of China. To address the possible transmissions and the relationships to the 7th pandemic strains of those 3 outbreaks, we sequenced the whole genomes of the outbreak isolates and compared with the global isolates from the 7th pandemic. The three outbreaks in this study were caused by a cluster of V. cholerae in clade 3.B which is parallel to the clade 3.C that was transmitted from Nepal to Haiti and caused an outbreak in 2010. Pan-genome analysis provided additional evolution information on the mobile element and acquired multiple antibiotic resistance genes. We suggested that clade 3.B should be monitored because the multiple antibiotic resistant characteristics of this clade and the 'amplifier' function of China in the global transmission of current Cholera pandemic. We also show that dedicated whole genome sequencing analysis provided more information than the previous techniques and should be applied in the disease surveillance networks.

  17. The Transmission and Antibiotic Resistance Variation in a Multiple Drug Resistance Clade of Vibrio cholerae Circulating in Multiple Countries in Asia.

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    Bo Pang

    Full Text Available Vibrio cholerae has caused massive outbreaks and even trans-continental epidemics. In 2008 and 2010, at least 3 remarkable cholera outbreaks occurred in Hainan, Anhui and Jiangsu provinces of China. To address the possible transmissions and the relationships to the 7th pandemic strains of those 3 outbreaks, we sequenced the whole genomes of the outbreak isolates and compared with the global isolates from the 7th pandemic. The three outbreaks in this study were caused by a cluster of V. cholerae in clade 3.B which is parallel to the clade 3.C that was transmitted from Nepal to Haiti and caused an outbreak in 2010. Pan-genome analysis provided additional evolution information on the mobile element and acquired multiple antibiotic resistance genes. We suggested that clade 3.B should be monitored because the multiple antibiotic resistant characteristics of this clade and the 'amplifier' function of China in the global transmission of current Cholera pandemic. We also show that dedicated whole genome sequencing analysis provided more information than the previous techniques and should be applied in the disease surveillance networks.

  18. Associação de Vibrio cholerae com o zooplâncton de águas estuárias da Baía de São Marcos/São Luis - MA, Brasil Association between Vibrio cholerae and zooplankton of estuaries of São Marcos Bay/São Luis - MA, Brazil

    OpenAIRE

    Eloisa da Graça do Rosario Gonçalves; Maria José Saraiva Lopes; Eurípedes Gomes de Oliveira; Ernesto Hofer

    2004-01-01

    Foi investigado, no período de outubro de 1997 a outubro de 1998, a possível associação de Vibrio cholerae com o zooplâncton dos estuários dos rios Anil e Bacanga, em São Luis - MA, Brasil, a presença da forma viável, mas não cultivável de Vibrio cholerae O1 e a correlação entre pH, salinidade e temperatura da água com a sobrevivência da bactéria. Amostras de zooplâncton foram coletadas em dois pontos fixos em cada estuário. O método clássico de isolamento e imunofluorescência direta foram em...

  19. Cyclo(valine-valine) inhibits Vibrio cholerae virulence gene expression.

    Science.gov (United States)

    Vikram, Amit; Ante, Vanessa M; Bina, X Renee; Zhu, Qin; Liu, Xinyu; Bina, James E

    2014-06-01

    Vibrio cholerae has been shown to produce a cyclic dipeptide, cyclo(phenylalanine-proline) (cFP), that functions to repress virulence factor production. The objective of this study was to determine if heterologous cyclic dipeptides could repress V. cholerae virulence factor production. To that end, three synthetic cyclic dipeptides that differed in their side chains from cFP were assayed for virulence inhibitory activity in V. cholerae. The results revealed that cyclo(valine-valine) (cVV) inhibited virulence factor production by a ToxR-dependent process that resulted in the repression of the virulence regulator aphA. cVV-dependent repression of aphA was found to be independent of known aphA regulatory genes. The results demonstrated that V. cholerae was able to respond to exogenous cyclic dipeptides and implicated the hydrophobic amino acid side chains on both arms of the cyclo dipeptide scaffold as structural requirements for inhibitory activity. The results further suggest that cyclic dipeptides have potential as therapeutics for cholera treatment.

  20. On the probability of extinction of the Haiti cholera epidemic

    Science.gov (United States)

    Bertuzzo, Enrico; Finger, Flavio; Mari, Lorenzo; Gatto, Marino; Rinaldo, Andrea

    2014-05-01

    Nearly 3 years after its appearance in Haiti, cholera has already exacted more than 8,200 deaths and 670,000 reported cases and it is feared to become endemic. However, no clear evidence of a stable environmental reservoir of pathogenic Vibrio cholerae, the infective agent of the disease, has emerged so far, suggesting that the transmission cycle of the disease is being maintained by bacteria freshly shed by infected individuals. Thus in principle cholera could possibly be eradicated from Haiti. Here, we develop a framework for the estimation of the probability of extinction of the epidemic based on current epidemiological dynamics and health-care practice. Cholera spreading is modelled by an individual-based spatially-explicit stochastic model that accounts for the dynamics of susceptible, infected and recovered individuals hosted in different local communities connected through hydrologic and human mobility networks. Our results indicate that the probability that the epidemic goes extinct before the end of 2016 is of the order of 1%. This low probability of extinction highlights the need for more targeted and effective interventions to possibly stop cholera in Haiti.

  1. Chemoproteomic profiling of host and pathogen enzymes active in cholera.

    Science.gov (United States)

    Hatzios, Stavroula K; Abel, Sören; Martell, Julianne; Hubbard, Troy; Sasabe, Jumpei; Munera, Diana; Clark, Lars; Bachovchin, Daniel A; Qadri, Firdausi; Ryan, Edward T; Davis, Brigid M; Weerapana, Eranthie; Waldor, Matthew K

    2016-04-01

    Activity-based protein profiling (ABPP) is a chemoproteomic tool for detecting active enzymes in complex biological systems. We used ABPP to identify secreted bacterial and host serine hydrolases that are active in animals infected with the cholera pathogen Vibrio cholerae. Four V. cholerae proteases were consistently active in infected rabbits, and one, VC0157 (renamed IvaP), was also active in human choleric stool. Inactivation of IvaP influenced the activity of other secreted V. cholerae and rabbit enzymes in vivo, and genetic disruption of all four proteases increased the abundance of intelectin, an intestinal lectin, and its binding to V. cholerae in infected rabbits. Intelectin also bound to other enteric bacterial pathogens, suggesting that it may constitute a previously unrecognized mechanism of bacterial surveillance in the intestine that is inhibited by pathogen-secreted proteases. Our work demonstrates the power of activity-based proteomics to reveal host-pathogen enzymatic dialog in an animal model of infection. PMID:26900865

  2. Environmental reservoirs and mechanisms of persistence of Vibrio cholerae

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    Carla eLutz

    2013-12-01

    Full Text Available It is now well accepted that Vibrio cholerae, the causative agent of the water-borne disease cholera, is acquired from environmental sources where it persists between outbreaks of the disease. Recent advances in molecular technology have demonstrated that this bacterium could be detected in areas where it had not been isolated from before, indicating a much broader, global distribution of this bacterium rather than specifically within regions where cholera is endemic. The environmental persistence of V. cholerae in the aquatic environment can be attributed to multiple intra- and interspecific strategies such as responsive gene regulation and biofilm formation on biotic and abiotic surfaces, as well as interactions with a multitude of other organisms. This review will discuss some of the mechanisms that enable the persistence of the bacterium in the sometimes hostile environment. In particular, we will discuss how V. cholerae can survive stressors such as starvation, temperature and salinity fluctuations as well as how the organism persists under constant predation by heterotrophic protists.

  3. Detection of Vibrio parahaemolyticus and Vibrio cholerae in oyster, Crassostrea rhizophorae, collected from a natural nursery in the Cocó river estuary, Fortaleza, Ceará, Brazil Isolamento de Vibrio parahaemolyticus e Vibrio cholerae em ostras, Crassostrea rhizophorae, coletadas em um criadouro natural no estuário do rio Cocó, Fortaleza, Ceará, Brasil

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    Oscarina Viana de Sousa

    2004-04-01

    Full Text Available Oysters are edible organisms that are often ingested partially cooked or even raw, presenting therefore a very high risk to the consumers' health, especially in tropical regions. The presence of Vibrio cholerae and Vibrio parahaemolyticus in oysters sampled at an estuary in the Brazilian northeastern region was studied, with 300 oysters tested through an 8-months period. The salinity of the water at the sampling point varied between 3% and 27‰. V. cholerae was the most frequently detected species (33.3% of the samples, and of the 22 V. cholerae isolates, 20 were identified as non-O1/non-O139, with two of the colonies presenting a rough surface and most of remaining ones belonging to the Heiberg II fermentation group. V. parahaemolyticus was isolated from just one of the samples. Other bacteria such as Providencia spp., Klebsiella spp. and Morganella morganii were also isolated.As ostras são alimentos marinhos freqüentemente ingeridos crus ou parcialmente cozidos. Por esta razão, o risco para a saúde dos consumidores desses produtos é muito elevado, principalmente, quando são de regiões tropicais. Foi estudada a presença de Vibrio cholerae e Vibrio parahaemolyticus em ostras de um estuário na região Nordeste do Brasil. Trezentas ostras foram analisadas, em um período de 8 meses. A salinidade da água, no local de coleta, variou de 3 a 27‰. V. cholerae foi o vibrio mais freqüentemente detectado (33,3% das amostras. Dos 22 isolados, 20 foram identificados como V. cholerae não-O1/não-O139, duas delas apresentando forma rugosa sendo a maioria das demais pertencente ao tipo fermentativo Heiberg II. V. parahaemolyticus foi isolado em apenas umas das coletas. Foram, também, identificadas nas amostras isolados de Providencia spp., Klebsiella spp., Proteus spp. e Morganella morganii.

  4. Conjugated Linoleic Acid Reduces Cholera Toxin Production In Vitro and In Vivo by Inhibiting Vibrio cholerae ToxT Activity.

    Science.gov (United States)

    Withey, Jeffrey H; Nag, Drubhajyoti; Plecha, Sarah C; Sinha, Ritam; Koley, Hemanta

    2015-12-01

    The severe diarrheal disease cholera is endemic in over 50 countries. Current therapies for cholera patients involve oral and/or intravenous rehydration, often combined with the use of antibiotics to shorten the duration and intensity of the disease. However, as antibiotic resistance increases, treatment options will become limited. Linoleic acid has been shown to be a potent negative effector of V. cholerae virulence that acts on the major virulence transcription regulator protein, ToxT, to inhibit virulence gene expression. ToxT activates transcription of the two major virulence factors required for disease, cholera toxin (CT) and toxin-coregulated pilus (TCP). A conjugated form of linoleic acid (CLA) is currently sold over the counter as a dietary supplement and is generally recognized as safe by the U.S. Food and Drug Administration. This study examined whether CLA could be used as a new therapy to reduce CT production, which, in turn, would decrease disease duration and intensity in cholera patients. CLA could be used in place of traditional antibiotics and would be very unlikely to generate resistance, as it affects only virulence factor production and not bacterial growth or survival.

  5. Cholera in Ecuador: Current relevance of past lessons learnt

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    S S Malavade

    2011-01-01

    Full Text Available This report analyses the trends in the cholera epidemic that hit Ecuador in 1991. The study is based on personal experiences and analysis of epidemiological databases from the Ministry of Public Health of Ecuador. The number of cases and initial attack rates in an immunologically naive population are described by province. An analysis of the Andean and coastal cholera patterns of transmission are described along with its associated risk factors. The logistical, environmental, and socio-cultural risk factors prevalent during the epidemic and the control measures implemented are also reviewed. Also, the role of the epidemic in the development of the public health and healthcare resources in Ecuador is discussed here. Current data indicate favorable conditions for another outbreak of cholera in Ecuador. In view of the existing risk factors, new strategies are proposed to prevent such an epidemic in the future.

  6. Cholera Surveillance in Siming District of Xiamen, 2005 -2010%2005-2010年厦门市思明区霍乱监测结果分析

    Institute of Scientific and Technical Information of China (English)

    陈沁; 阮菁如; 刘建明; 苏志寿

    2011-01-01

    目的 为了解思明区食品及外环境水体中霍乱弧菌的污染状况,为制订霍乱防控策略提供依据.方法根据2005-2010年《福建省霍乱监测方案》要求,对思明区食品及外环境水体开展霍乱弧菌监测.结果共采集各类标本4861份,检出霍乱弧菌78份,总阳性率1.60%.食品检测3517份,检出霍乱弧菌73份,阳性率2.08%,各类食品以蛙类阳性率(12.30%)最高,其次为甲壳类(2.69%)、鱼类(2.18%).水体检测1344份,检出霍乱弧茵5份,阳性率0.43%.检出的霍乱弧菌中,45例为01群稻叶型,33例为01群小川型,经霍乱毒素基因检测均为非产毒株.结论思明区水产品中的霍乱弧菌污染程度较高,应加强对蛙类食品的监测和管理.%Objective To investigate the contamination situation of Vibrio Cholera in foods and external environmental water bodies in Siming district, and provide scientific evidence for the prevention and control of cholera. Methods Foods and external environmental water bodies were monitored for Vibrio Cholera in Siming district according to the requirements of Cholera Surveillance Program In Fujian Province (2005 -2010). Results A total of 4 861 specimens of different types were collected, and 78 samples with Vibrio Cholera were detected. The total positive rate was 1. 60%. In which 3 517 foods were collected, and Vibrio Cholera were detected in 73 samples, with positive rate 2.08%. The highest positive rate was from frog species (12. 30% ) in food, fol-lowed by crustaceans (2. 69% ) , and fishes (2. 18% ); 1 344 water bodies were collected, and 5 samples with Vibrio Cholera were detected, with positive rate 0.43%. Among the detections, Vibrio Cholera in 45 samples were Inaba type and in 33 samples were Ogawa type, and all the bacterial strains were non - toxigenic strains according to the cholera toxin gene detection. Conclusion The contamination levels of Vibrio Cholera in the aquatic products are higher than expected, the

  7. Spatial and environmental connectivity analysis in a cholera vaccine trial.

    Science.gov (United States)

    Emch, Michael; Ali, Mohammad; Root, Elisabeth D; Yunus, Mohammad

    2009-02-01

    This paper develops theory and methods for vaccine trials that utilize spatial and environmental information. Satellite imagery is used to identify whether households are connected to one another via water bodies in a study area in rural Bangladesh. Then relationships between neighborhood-level cholera vaccine coverage and placebo incidence and neighborhood-level spatial variables are measured. The study hypothesis is that unvaccinated people who are environmentally connected to people who have been vaccinated will be at lower risk compared to unvaccinated people who are environmentally connected to people who have not been vaccinated. We use four datasets including: a cholera vaccine trial database, a longitudinal demographic database of the rural population from which the vaccine trial participants were selected, a household-level geographic information system (GIS) database of the same study area, and high resolution Quickbird satellite imagery. An environmental connectivity metric was constructed by integrating the satellite imagery with the vaccine and demographic databases linked with GIS. The results show that there is a relationship between neighborhood rates of cholera vaccination and placebo incidence. Thus, people are indirectly protected when more people in their environmentally connected neighborhood are vaccinated. This result is similar to our previous work that used a simpler Euclidean distance neighborhood to measure neighborhood vaccine coverage [Ali, M., Emch, M., von Seidlein, L., Yunus, M., Sack, D. A., Holmgren, J., et al. (2005). Herd immunity conferred by killed oral cholera vaccines in Bangladesh. Lancet, 366(9479), 44-49]. Our new method of measuring environmental connectivity is more precise since it takes into account the transmission mode of cholera and therefore this study validates our assertion that the oral cholera vaccine provides indirect protection in addition to direct protection.

  8. Influence of climate factors on Vibrio cholerae dynamics in the Pearl River estuary, South China.

    Science.gov (United States)

    Yue, Yujuan; Gong, Jianhua; Wang, Duochun; Kan, Biao; Li, Baisheng; Ke, Changwen

    2014-06-01

    Current research has seldom focused on the quantitative relationships between Vibrio cholerae (V. cholerae) and climate factors owing to the complexities and high cost of field observation in the aquatic environment. This study has focused on the relationships between V. cholerae and climate factors based on linear regression method and data partition method. Data gathered from 2008 to 2009 in the Pearl River estuary, South China, were adopted. Positive rate of V. cholerae was correlated closely with monthly climate factors of water temperature and air temperature, respectively in 2009. Quarterly data analysis from 2008 to 2009 showed that there existed seasonal characteristic for V. cholerae. Positive rate of V. cholerae was correlated positively with quarterly climate factors of land surface temperature, pH, water temperature, air temperature and rainfall, respectively and negatively with quarterly air pressure. Partition data analysis in 2009 showed that there existed geography region characteristic for V. cholerae. V. cholerae dynamics was closely correlated to climate factors in the downstream area. However, it was more greatly affected by human geography factors in the urban area. Positive annual rate of V. cholerae was higher in the downstream area than in the urban area both in 2008 and 2009. At last, a cellular automaton model was used to simulate V. cholerae diffusion downstream, and the distribution of V. cholerae obtained from this model was similar to that obtained from the field observations.

  9. An intracellular replication niche for Vibrio cholerae in the amoeba Acanthamoeba castellanii.

    Science.gov (United States)

    Van der Henst, Charles; Scrignari, Tiziana; Maclachlan, Catherine; Blokesch, Melanie

    2016-04-01

    Vibrio cholerae is a human pathogen and the causative agent of cholera. The persistence of this bacterium in aquatic environments is a key epidemiological concern, as cholera is transmitted through contaminated water. Predatory protists, such as amoebae, are major regulators of bacterial populations in such environments. Therefore, we investigated the interaction between V. cholerae and the amoeba Acanthamoeba castellanii at the single-cell level. We observed that V. cholerae can resist intracellular killing. The non-digested bacteria were either released or, alternatively, established a replication niche within the contractile vacuole of A. castellanii. V. cholerae was maintained within this compartment even upon encystment. The pathogen ultimately returned to its aquatic habitat through lysis of A. castellanii, a process that was dependent on the production of extracellular polysaccharide by the pathogen. This study reinforces the concept that V. cholerae is a facultative intracellular bacterium and describes a new host-pathogen interaction.

  10. Field evaluation of environmental sanitation measures against cholera.

    Science.gov (United States)

    Azurin, J C; Alvero, M

    1974-01-01

    Data obtained in a controlled field study over 5 years in 4 communities showed that the provision of sanitary facilities for human waste disposal can reduce the incidence of cholera by as much as 68%, while the provision of a safe water supply can decrease it by 73%. Where both toilets and water supplies are provided, the incidence can be reduced by as much as 76%. There was evidence that cholera infection gaining access to communities with these facilities tends to spread less and produce fewer secondary cases than in a community where such facilities are not provided. PMID:4549038

  11. Field evaluation of environmental sanitation measures against cholera*

    Science.gov (United States)

    Azurin, J. C.; Alvero, M.

    1974-01-01

    Data obtained in a controlled field study over 5 years in 4 communities showed that the provision of sanitary facilities for human waste disposal can reduce the incidence of cholera by as much as 68%, while the provision of a safe water supply can decrease it by 73%. Where both toilets and water supplies are provided, the incidence can be reduced by as much as 76%. There was evidence that cholera infection gaining access to communities with these facilities tends to spread less and produce fewer secondary cases than in a community where such facilities are not provided. PMID:4549038

  12. Occurrence of Vibrio Parahaemolyticus, Vibrio Cholerae and Vibrio Vulnificus in the Clam Ruditapes Philippinarum (Adams & Reeve, 1850) from Emilia Romagna and Sardinia, Italy

    Science.gov (United States)

    Passalacqua, Pier Luca; Zavatta, Emanuele; Bignami, Giorgia; Serraino, Andrea

    2016-01-01

    Marine vibrios, Vibrio parahaemolyticus, V. vulnificus and V. cholerae are responsible of the majority of food-borne human infections by consumption of bivalve shellfish. The aim of the present study was to ascertain the occurrence of these bacteria, and their potential pathogenicity, in the Manila clam R. philippinarum from Emilia Romagna (ER) and Sardinia (SR) regions, Italy. Isolation was performed on CHROMagarTM vibrio with subculture on (thiosulfate-citrate-bile salts-sucrose) Agar and m-modified-cellobiose-polymyxin b-colistin (-CPC) Agar. Suspected strains were purified, biochemically characterized and genotyped by simplex polymerase chain reaction (PCR) for the specie-specific and pathogenic gene markers: V. parahaemolyticus (toxRP, tdh and trh); V. vulnificus (vvhA, hsp, vcgC, vcgE, CPS operon allele 1, CPS operon allele 2, 16s-rRNA operon allele A, 16s-rRNA operon allele B; V. cholerae (toxRC, hlya, tcpI, tcpA, ctxA, ctxB, stn/sto). Moreover a multiplex PCR was applied to the SR bivalve shellfish, for the simultaneous detection of the three targets directly on homogenate samples, targeting the species-specific gene for V. cholerae (toxRC), V. parahaemolyticus (toxRP) and V. vulnificus (vvhA). As a result of phenotyping and genotyping of isolates, bivalve shellfish from ER resulted positive for V. parahaemolyticus (27.8%) and V. vulnificus (10.1%), but negative for V. cholerae. Shellfish from SR resulted positive for V. parahaemolyticus (30.3%), V. vulnificus (6.1%) and V. cholerae (3%). No significant differences emerged between the two areas (P>0.05).

  13. O1群霍乱弧菌新变异株的研究进展

    Institute of Scientific and Technical Information of China (English)

    沈定树; 余素飞; 历世笑

    2012-01-01

    O1群霍乱弧菌( Vibrio cholerae)是霍乱的病原体[1].根据其"O"抗原的不同可分为200多个血清群[2].根据其表型差异,O1群霍乱弧菌又可分为古典型(Classical biotypeV.cholerae,CVC)和El Tor型(El Tor biotype V.cholerae,EVC)2个生物型.古典型与El Tor型不仅在表型和基因特性上不同,在其致病性以及在宿主体内的感染模式与菌体残留时间也有所不同.例如,El Tor型通常是无症状感染,死亡率相对较低,在人宿主体内存活率高,在宿主与宿主间的传播较快;El Tor型还具有很强的生态适应性,在适宜环境中可持续存在时间长[3,4].一般来说,古典型引起的霍乱比El Tor型其后果更严重.近年来在非洲和亚洲的许多国家中分离出霍乱弧菌变异菌株,这些变异菌株中隐藏有古典型与El Tor型的某些表型和基因型.Na- Ubol等[5]对泰国1986年至2009年间临床分离的O1群霍乱弧菌的基因分析特征表明,自1986年开始泰国的霍乱流行是以典型El Tor型与杂种生物型联合循环(co- circulating),至1992年是典型E1 Tor型最后一次分离出.

  14. Cholera vaccination campaign contributes to improved knowledge regarding cholera and improved practice relevant to waterborne disease in rural Haiti.

    Directory of Open Access Journals (Sweden)

    Omowunmi Aibana

    2013-11-01

    Full Text Available BACKGROUND: Haiti's cholera epidemic has been devastating partly due to underlying weak infrastructure and limited clean water and sanitation. A comprehensive approach to cholera control is crucial, yet some have argued that oral cholera vaccination (OCV might result in reduced hygiene practice among recipients. We evaluated the impact of an OCV campaign on knowledge and health practice in rural Haiti. METHODOLOGY/PRINCIPAL FINDINGS: We administered baseline surveys on knowledge and practice relevant to cholera and waterborne disease to every 10th household during a census in rural Haiti in February 2012 (N = 811. An OCV campaign occurred from May-June 2012 after which we administered identical surveys to 518 households randomly chosen from the same region in September 2012. We compared responses pre- and post-OCV campaign. Post-vaccination, there was improved knowledge with significant increase in percentage of respondents with ≥ 3 correct responses on cholera transmission mechanisms (odds ratio[OR] 1.91; 95% confidence interval[CI] 1.52-2.40, preventive methods (OR 1.83; 95% CI 1.46-2.30, and water treatment modalities (OR 2.75; 95% CI 2.16-3.50. Relative to pre-vaccination, participants were more likely post-OCV to report always treating water (OR 1.62; 95% CI 1.28-2.05. Respondents were also more likely to report hand washing with soap and water >4 times daily post-vaccine (OR 1.30; 95% CI 1.03-1.64. Knowledge of treating water as a cholera prevention measure was associated with practice of always treating water (OR 1.47; 95% CI 1.14-1.89. Post-vaccination, knowledge was associated with frequent hand washing (OR 2.47; 95% CI 1.35-4.51. CONCLUSION: An OCV campaign in rural Haiti was associated with significant improvement in cholera knowledge and practices related to waterborne disease. OCV can be part of comprehensive cholera control and reinforce, not detract from, other control efforts in Haiti.

  15. Vibrio vulnificus phage PV94 is closely related to temperate phages of V. cholerae and other Vibrio species.

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    Mark Pryshliak

    Full Text Available BACKGROUND: Vibrio vulnificus is an important pathogen which can cause serious infections in humans. Yet, there is limited knowledge on its virulence factors and the question whether temperate phages might be involved in pathogenicity, as is the case with V. cholerae. Thus far, only two phages (SSP002 and VvAW1 infecting V. vulnificus have been genetically characterized. These phages were isolated from the environment and are not related to Vibrio cholerae phages. The lack of inf