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Sample records for marker cholera toxin

  1. Radiolabelling of cholera toxin

    International Nuclear Information System (INIS)

    Santos, R.G.; Neves, Nicoli M.J.; Abdalla, L.F.; Brandao, R.L.; Etchehebehere, L.; Lima, M.E. de; Nicoli, J.R.

    1999-01-01

    Binding of cholera toxin to ganglioside receptors of enterocyte microvilli catalyzes the activation of adenylate cyclase causing a rise in cAMP which final result is a copious diarrhea. Saccharomyces boulardii, a nonpathogenic yeast has been used to prevent diarrhea. Although the antidiarrheic properties of S. boulardii are widely recognized, this yeast has been used on empirical basis, and the mechanism of this protective effect is unknown. The addition of cholera toxin to S. boulardii induces the raising of cAMP that triggers the activation of neutral trehalase. This suggests that toxin specifically binding to cells, is internalized and active the protein phosphorylation cascade. Our objective is labeling the cholera toxin to verify the presence of binding sites on yeast cell surfaces for the cholera toxin. Cholera toxin was radiolabelled with Na 125 I by a chloramine-T method modified from Cuatrecasas and Griffiths et alii. The 125 I-Cholera toxin showed a specific radioactivity at about 1000 cpm/fmol toxin. Biological activity of labeled cholera toxin measured by trehalase activation was similar to the native toxin. (author)

  2. Radiolabelling of cholera toxin

    Energy Technology Data Exchange (ETDEWEB)

    Santos, R.G.; Neves, Nicoli M.J. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN), Belo Horizonte, MG (Brazil); Abdalla, L.F.; Brandao, R.L.; Etchehebehere, L. [Ouro Preto Univ., MG (Brazil). Escola de Farmacia. Lab. de Fisiologia e Bioquimica de Microorganismos; Lima, M.E. de [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Bioquimica e Imunologia; Nicoli, J.R. [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Microbiologia

    1999-11-01

    Binding of cholera toxin to ganglioside receptors of enterocyte microvilli catalyzes the activation of adenylate cyclase causing a rise in cAMP which final result is a copious diarrhea. Saccharomyces boulardii, a nonpathogenic yeast has been used to prevent diarrhea. Although the antidiarrheic properties of S. boulardii are widely recognized, this yeast has been used on empirical basis, and the mechanism of this protective effect is unknown. The addition of cholera toxin to S. boulardii induces the raising of cAMP that triggers the activation of neutral trehalase. This suggests that toxin specifically binding to cells, is internalized and active the protein phosphorylation cascade. Our objective is labeling the cholera toxin to verify the presence of binding sites on yeast cell surfaces for the cholera toxin. Cholera toxin was radiolabelled with Na {sup 125} I by a chloramine-T method modified from Cuatrecasas and Griffiths et alii. The {sup 125} I-Cholera toxin showed a specific radioactivity at about 1000 cpm/fmol toxin. Biological activity of labeled cholera toxin measured by trehalase activation was similar to the native toxin. (author) 5 refs., 3 figs.; e-mail: nevesmj at urano.cdtn.br

  3. Crystallization of isoelectrically homogeneous cholera toxin

    International Nuclear Information System (INIS)

    Spangler, B.D.; Westbrook, E.M.

    1989-01-01

    Past difficulty in growing good crystals of cholera toxin has prevented the study of the crystal structure of this important protein. The authors have determined that failure of cholera toxin to crystallize well has been due to its heterogeneity. They have now succeeded in overcoming the problem by isolating a single isoelectric variant of this oligomeric protein (one A subunit and five B subunits). Cholera toxin purified by their procedure readily forms large single crystals. The crystal form has been described previously. They have recorded data from native crystals of cholera toxin to 3.0-angstrom resolution with our electronic area detectors. With these data, they have found the orientation of a 5-fold symmetry axis within these crystals, perpendicular to the screw dyad of the crystal. They are now determining the crystal structure of cholera toxin by a combination of multiple heavy-atom isomorphous replacement and density modification techniques, making use of rotational 5-fold averaging of the B subunits

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

    International Nuclear Information System (INIS)

    Peterson, J.W.; Berg, W.D. Jr.; Coppenhaver, D.H.

    1987-01-01

    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 [ 3 H] 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 [ 35 S] 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

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

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    Jussi A. O. Meriluoto

    2012-06-01

    Full Text Available Cholera remains a serious health problem, especially in developing countries where basic hygiene standards are not met. The symptoms of cholera are caused by cholera toxin, an enterotoxin, which is produced by the bacterium Vibrio cholerae. We have recently shown that human probiotic bacteria are capable of removing cyanobacterial toxins from aqueous solutions. In the present study we investigate the ability of the human probiotic bacteria, Lactobacillus rhamnosus strain GG (ATCC 53103 and Bifidobacterium longum 46 (DSM 14583, to remove cholera toxin from solution in vitro. Lactobacillus rhamnosus strain GG and Bifidobacterium longum 46 were able to remove 68% and 59% of cholera toxin from aqueous solutions during 18 h of incubation at 37 °C, respectively. The effect was dependent on bacterial concentration and L. rhamnosus GG was more effective at lower bacterial concentrations. No significant effect on cholera toxin concentration was observed when nonviable bacteria or bacterial supernatant was used.

  6. Actions of cholera toxin and the prevention and treatment of cholera

    Science.gov (United States)

    Holmgren, Jan

    1981-07-01

    The drastic intestinal secretion of fluid and electrolytes that is characteristic of cholera is the result of reasonably well understood cellular and biochemical actions of the toxin secreted by Vibrio cholerae. Based on this understanding it is possible to devise new techniques for the treatment and prophylaxis of cholera to complement those based on fluid replacement therapy and sanitation.

  7. Evaluation of Cholera Toxin Expression in Acidic, Alkaline and Neutral Conditions

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    Narges Rahimi

    2015-02-01

    Full Text Available Background: Cholera is a severe disease which is caused by Vibrio cholerae and it is typically transmitted by either contaminated food or water particularly in developing countries. The most important virulence factor of this bacterium is an enterotoxin called cholera toxin which is a protein complex secreted by the Vibrio cholerae. Objectives: In this project, we determined the production of cholera toxin at different pH values. Materials and Methods: Two standard strain of Vibrio cholerae O1 biovar EL Tor N16961 and Vibrio cholerae O1 biovar Classic ATCC 14035 were used. After overnight cultivation of both the strains the total mRNA extracted and converted to total cDNA. Results: By Relative Real-Time PCR analysis the most cholera toxin production in classical and El Tor strains was at pH 8.5 and 8, respectively. Conclusions: Therefore, We may conclude that use of acidic diet will help in reduction of cholera toxin production.

  8. Cholera toxin can catalyze ADP-ribosylation of cytoskeletal proteins

    International Nuclear Information System (INIS)

    Kaslow, H.R.; Groppi, V.E.; Abood, M.E.; Bourne, H.R.

    1981-01-01

    Cholera toxin catalyzes transfer of radiolabel from [ 32 P]NAD + to several peptides in particulate preparations of human foreskin fibroblasts. Resolution of these peptides by two-dimensional gel electrophoresis allowed identification of two peptides of M/sub r/ = 42,000 and 52,000 as peptide subunits of a regulatory component of adenylate cyclase. The radiolabeling of another group of peptides (M/sub r/ = 50,000 to 65,000) suggested that cholera toxin could catalyze ADP-ribosylation of cytoskeletal proteins. This suggestion was confirmed by showing that incubation with cholera toxin and [ 32 P]NAD + caused radiolabeling of purified microtubule and intermediate filament proteins

  9. Changes in intestinal fluid and mucosal immune responses to cholera toxin in Giardia muris infection and binding of cholera toxin to Giardia muris trophozoites.

    Science.gov (United States)

    Ljungström, I; Holmgren, J; Svennerholm, A M; Ferrante, A

    1985-10-01

    The effect of Giardia muris infection on the diarrheal response and gut mucosal antibody response to cholera toxin was examined in mice. The results obtained showed that the fluid accumulation in intestinal loops exposed to cholera toxin was increased in mice infected with a low number (5 X 10(4) ) of G. muris cysts compared with the response in noninfected mice. This effect was associated with a marked reduction in absorption of oral rehydration fluid from the intestine. In contrast, mice infected with a high dose (2 X 10(5) ) of cysts showed a marked decrease in fluid accumulation in response to the toxin. This decrease might be related to the finding that both G. muris and Giardia lamblia trophozoites can bind significant amounts of cholera toxin. Evidence is presented which suggests that the gut mucosal antibody response, mainly immunoglobulin A but also immunoglobulin G, to an immunization course with perorally administered cholera toxin was depressed in mice infected with G. muris. The reduction in antibody levels was particularly evident when the primary immunization was made very early after infection. The serum antitoxin antibodies to the oral immunization with cholera toxin were, however, not affected. Likewise, the delayed-type hypersensitivity response against sheep erythrocytes in animals primed subcutaneously with sheep erythrocytes was not modified during the course of G. muris infection.

  10. ADP-ribosylation by cholera toxin: functional analysis of a cellular system that stimulates the enzymic activity of cholera toxin fragment A1

    International Nuclear Information System (INIS)

    Gill, D.M.; Coburn, J.

    1987-01-01

    The authors have clarified relationships between cholera toxin, cholera toxin substrates, a membrane protein S that is required for toxin activity, and a soluble protein CF that is needed for the function of S. The toxin has little intrinsic ability to catalyze ADP-ribosylations unless it encounters the active form of the S protein, which is S liganded to GTP or to a GTP analogue. In the presence of CF, S x GTP forms readily, though reversibly, but a more permanent active species, S-guanosine 5'-O-(3-thiotriphosphate) (S x GTPγS), forms over a period of 10-15 min at 37 0 C. Both guanosine 5'-O-(2-thiodiphosphate) and GTP block this quasi-permanent activation. Some S x GTPγS forms in membranes that are exposed to CF alone and then to GTPγS, with a wash in between, and it is possible that CF facilitates a G nucleotide exchange. S x GTPγS dissolved by nonionic detergents persists in solution and can be used to support the ADP-ribosylation of nucleotide-free substrates. In this circumstance, added guanyl nucleotides have no further effect. This active form of S is unstable, especially when heated, but the thermal inactivation above 45 0 C is decreased by GTPγS. Active S is required equally for the ADP-ribosylation of all of cholera toxin's protein substrates, regardless of whether they bind GTP or not. They suggest that active S interacts directly with the enzymic A 1 fragments of cholera toxin and not with any toxin substrate. The activation and activity of S are independent of the state, or even the presence, of adenylate cyclase and seem to be involved with the cyclase system only via cholera toxin. S is apparently not related by function to certain other GTP binding proteins, including p21/sup ras/, and appears to be a new GTP binding protein whose physiologic role remains to be identified

  11. ADP-ribosylation of membrane components by pertussis and cholera toxin

    International Nuclear Information System (INIS)

    Ribeiro-Neto, F.A.P.; Mattera, F.; Hildebrandt, J.D.; Codina, J.; Field, J.B.; Birnbaumer, L.; Sekura, R.D.

    1985-01-01

    Pertussis and cholera toxins are important tools to investigate functional and structural aspects of the stimulatory (N/sub s/) and inhibitory (N/sub i/) regulatory components of adenylyl cyclase. Cholera toxin acts on N/sub s/ by ADP-ribosylating its α/sub s/ subunit; pertussis toxin acts on N/sub i/ by ADP-ribosylating its α; subunit. By using [ 32 P]NAD + and determining the transfer of its [ 32 P]ADP-ribose moiety to membrane components, it is possible to obtain information on N/sub s/ and N/sub i/. A set of protocols is presented that can be used to study simultaneously and comparatively the susceptibility of N/sub s/ and N/sub i/ to be ADP-ribosylated by cholera and pertussis toxin

  12. Detection of cholera (ctx) and zonula occludens (zot) toxin genes in Vibrio cholerae O1, O139 and non-O1 strains.

    Science.gov (United States)

    Rivera, I G; Chowdhury, M A; Sanchez, P S; Sato, M I; Huq, A; Colwell, R R; Martins, M T

    1995-09-01

    Vibrio cholerae O1 and V. cholerae non-O1 strains isolated from environmental samples collected in São Paulo, Brazil, during cholera epidemics and pre-epidemic periods were examined for the presence of toxin genes. V. cholerae O1 strains isolated from clinical samples in Peru and Mexico, and V. cholerae O139 strains from India were also examined for the presence of ctx (cholera toxin gene) and zot (zonula occludens toxin gene) by polymerase chain reaction (PCR). A modified DNA-extraction method applied in this study yielded satisfactory recovery of genomic DNA from vibrios. Results showed that strains of V. cholerae O1 isolated during the preepidemic period were ctx (-)/zot (-) whereas strains isolated during the epidemic were ctx (+)/zot (+). All V. cholerae non-O1 strains tested in the study were ctx (-)/zot (-), whereas all V. cholerae O139 strains were ctx (+)/zot (+). Rapid detection of the virulence genes (ctx and zot) can be achieved by PCR and this can serve as an important tool in the epidemiology and surveillance of V. cholerae.

  13. Cholix Toxin, a Novel ADP-ribosylating Factor from Vibrio cholerae

    Energy Technology Data Exchange (ETDEWEB)

    Jorgensen, Rene; Purdy, Alexandra E.; Fieldhouse, Robert J.; Kimber, Matthew S.; Bartlett, Douglas H.; Merrill, A. Rod (Guelph); (NIH); (UCSD)

    2008-07-15

    The ADP-ribosyltransferases are a class of enzymes that display activity in a variety of bacterial pathogens responsible for causing diseases in plants and animals, including those affecting mankind, such as diphtheria, cholera, and whooping cough. We report the characterization of a novel toxin from Vibrio cholerae, which we call cholix toxin. The toxin is active against mammalian cells (IC50 = 4.6 {+-} 0.4 ng/ml) and crustaceans (Artemia nauplii LD50 = 10 {+-} 2 {mu}g/ml). Here we show that this toxin is the third member of the diphthamide-specific class of ADP-ribose transferases and that it possesses specific ADP-ribose transferase activity against ribosomal eukaryotic elongation factor 2. We also describe the high resolution crystal structures of the multidomain toxin and its catalytic domain at 2.1- and 1.25-{angstrom} resolution, respectively. The new structural data show that cholix toxin possesses the necessary molecular features required for infection of eukaryotes by receptor-mediated endocytosis, translocation to the host cytoplasm, and inhibition of protein synthesis by specific modification of elongation factor 2. The crystal structures also provide important insight into the structural basis for activation of toxin ADP-ribosyltransferase activity. These results indicate that cholix toxin may be an important virulence factor of Vibrio cholerae that likely plays a significant role in the survival of the organism in an aquatic environment.

  14. Molecular Dynamics Simulation of Cholera Toxin A-1 Polypeptide

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    Badshah Syed Lal

    2016-01-01

    Full Text Available A molecular dynamics (MD simulation study of the enzymatic portion of cholera toxin; cholera toxin A-1 polypeptide (CTA1 was performed at 283, 310 and 323 K. From total energy analysis it was observed that this toxin is stable thermodynamically and these outcomes were likewise confirmed by root mean square deviations (RMSD investigations. The Cα root mean square fluctuation (RMSF examinations revealed that there are a number of residues inside CTA1, which can be used as target for designing and synthesizing inhibitory drugs, in order to inactivate cholera toxin inside the human body. The fluctuations in the radius of gyration and hydrogen bonding in CTA1 proved that protein unfolding and refolding were normal routine phenomena in its structure at all temperatures. Solvent accessible surface area study identified the hydrophilic nature of the CTA1, and due to this property it can be a potential biological weapon. The structural identification (STRIDE algorithm for proteins was successfully used to determine the partially disordered secondary structure of CTA1. On account of this partially disordered secondary structure, it can easily deceive the proteolytic enzymes of the endoplasmic reticulum of host cells.

  15. Immunization with cholera toxin B subunit induces high-level protection in the suckling mouse model of cholera.

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    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.

  16. Nanomechanical detection of cholera toxin using microcantilevers functionalized with ganglioside nanodiscs

    Energy Technology Data Exchange (ETDEWEB)

    Tark, Soo-Hyun; Dravid, Vinayak P [Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208 (United States); Das, Aditi; Sligar, Stephen, E-mail: s-sligar@illinois.edu, E-mail: v-dravid@northwestern.edu [Department of Biochemistry and Chemistry, Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801 (United States)

    2010-10-29

    The label-free detection of cholera toxin is demonstrated using microcantilevers functionalized with ganglioside nanodiscs. The cholera toxin molecules bind specifically to the active membrane protein encased in nanodiscs, nanoscale lipid bilayers surrounded by an amphipathic protein belt, immobilized on the cantilever surface. The specific molecular binding results in cantilever deflection via the formation of a surface stress-induced bending moment. The nanomechanical cantilever response is quantitatively monitored by optical interference. The consistent and reproducible nanomechanical detection of cholera toxin in nanomolar range concentrations is demonstrated. The results validated with such a model system suggest that the combination of a microcantilever platform with receptor nanodiscs is a promising approach for monitoring invasive pathogens and other types of biomolecular detection relevant to drug discovery.

  17. Nanomechanical detection of cholera toxin using microcantilevers functionalized with ganglioside nanodiscs

    International Nuclear Information System (INIS)

    Tark, Soo-Hyun; Dravid, Vinayak P; Das, Aditi; Sligar, Stephen

    2010-01-01

    The label-free detection of cholera toxin is demonstrated using microcantilevers functionalized with ganglioside nanodiscs. The cholera toxin molecules bind specifically to the active membrane protein encased in nanodiscs, nanoscale lipid bilayers surrounded by an amphipathic protein belt, immobilized on the cantilever surface. The specific molecular binding results in cantilever deflection via the formation of a surface stress-induced bending moment. The nanomechanical cantilever response is quantitatively monitored by optical interference. The consistent and reproducible nanomechanical detection of cholera toxin in nanomolar range concentrations is demonstrated. The results validated with such a model system suggest that the combination of a microcantilever platform with receptor nanodiscs is a promising approach for monitoring invasive pathogens and other types of biomolecular detection relevant to drug discovery.

  18. [GM1-dot-EIA for the detection of toxin-producing Vibrio cholerae strains].

    Science.gov (United States)

    Markina, O V; Alekseeva, L P; Telesmanich, N R; Chemisova, O S; Akulova, M V; Markin, N V

    2011-05-01

    A new variant of enzyme immunoassay (EIA) has been developed on the basis of GM1 gangliosides to detect the toxin-producing Vibrio cholerae strains--GM1-dot-EIA. Experiments were run using a nitrocellulose membrane to bind GM1 gangliosides and polyclonal antitoxic serum to detect cholerogen. GM1-dot-EIA testing identified cholera toxin in 11 of 13 supernatants of V. cholerae eltor ctx(+) strains isolated from man and in 3 of 7 supernatants of V. cholerae eltor ctx(+) strains isolated from water. These data agree with those obtained in CM1-EIA. There was no reaction with the supernatants of other microorganisms. The sensitivity of the technique was 10 ng/ml. Thus, the simple and specific GM1-dot-EIA may be recommended to detect toxin-producing V cholerae strains isolated from man and water.

  19. Persistence of plasmids, cholera toxin genes, and prophage DNA in classical Vibrio cholerae O1.

    Science.gov (United States)

    Cook, W L; Wachsmuth, K; Johnson, S R; Birkness, K A; Samadi, A R

    1984-07-01

    Plasmid profiles, the location of cholera toxin subunit A genes, and the presence of the defective VcA1 prophage genome in classical Vibrio cholerae isolated from patients in Bangladesh in 1982 were compared with those in older classical strains isolated during the sixth pandemic and with those in selected eltor and nontoxigenic O1 isolates. Classical strains typically had two plasmids (21 and 3 megadaltons), eltor strains typically had no plasmids, and nontoxigenic O1 strains had zero to three plasmids. The old and new isolates of classical V. cholerae had two HindIII chromosomal digest fragments containing cholera toxin subunit A genes, whereas the eltor strains from Eastern countries had one fragment. The eltor strains from areas surrounding the Gulf of Mexico also had two subunit A gene fragments, which were smaller and easily distinguished from the classical pattern. All classical strains had 8 to 10 HindIII fragments containing the defective VcA1 prophage genome; none of the Eastern eltor strains had these genes, and the Gulf Coast eltor strains contained a different array of weakly hybridizing genes. These data suggest that the recent isolates of classical cholera in Bangladesh are closely related to the bacterial strain(s) which caused classical cholera during the sixth pandemic. These data do not support hypotheses that either the eltor or the nontoxigenic O1 strains are precursors of the new classical strains.

  20. Cholera toxin entry into pig enterocytes occurs via a lipid raft- and clathrin-dependent mechanism

    DEFF Research Database (Denmark)

    Hansen, Gert H; Dalskov, Stine-Mathilde; Rasmussen, Christina Rehné

    2005-01-01

    The small intestinal brush border is composed of lipid raft microdomains, but little is known about their role in the mechanism whereby cholera toxin gains entry into the enterocyte. The present work characterized the binding of cholera toxin B subunit (CTB) to the brush border and its internaliz......The small intestinal brush border is composed of lipid raft microdomains, but little is known about their role in the mechanism whereby cholera toxin gains entry into the enterocyte. The present work characterized the binding of cholera toxin B subunit (CTB) to the brush border and its...... accompanied the toxin internalization whereas no formation of caveolae was observed. CTB was strongly associated with the buoyant, detergent-insoluble fraction of microvillar membranes. Neither CTB's raft association nor uptake via clathrin-coated pits was affected by methyl-beta-cyclodextrin, indicating...... that membrane cholesterol is not required for toxin binding and entry. The ganglioside GM(1) is known as the receptor for CTB, but surprisingly the toxin also bound to sucrase-isomaltase and coclustered with this glycosidase in apical membrane pits. CTB binds to lipid rafts of the brush border...

  1. Effects of cholera toxin and isobutylmethylxanthine on growth of human fibroblasts

    International Nuclear Information System (INIS)

    Espinoza, B.; Wharton, W.

    1986-01-01

    Cholera toxin produced a dose-dependent decrease in the restimulation of G 0 /G 1 traverse in density-arrested human fibroblasts but did not inhibit the stimulation of cells arrested in G 0 after serum starvation at low density. In addition, cholera toxin did not inhibit the proliferation of sparse logarithmically growing human fibroblasts, even when low concentrations of the phosphodiesterase inhibitor isobutylmethylxanthine (IBMX) were also present. However, the final density to which sparse cells grew was limited by cholera toxin, when added either alone or together with low concentrations of IBMX. In contrast, high concentrations of the phosphodiesterase inhibitor alone produced a profound inhibition in the growth of sparse human fibrobasts. IBMX produced an inhibition both in the G 1 and in the G 2 phases of the cell cycle by a mechanism(s) that was not related to the magnitude of the increases in adenosine 3,5-cyclic monophosphate concentrations

  2. Diversity and distribution of cholix toxin, a novel ADP-ribosylating factor from Vibrio cholerae.

    Science.gov (United States)

    Purdy, Alexandra E; Balch, Deborah; Lizárraga-Partida, Marcial Leonardo; Islam, Mohammad Sirajul; Martinez-Urtaza, Jaime; Huq, Anwar; Colwell, Rita R; Bartlett, Douglas H

    2010-02-01

    Non-toxigenic non-O1, non-O139 Vibrio cholerae strains isolated from both environmental and clinical settings carry a suite of virulence factors aside from cholera toxin. Among V. cholerae strains isolated from coastal waters of southern California, this includes cholix toxin, an ADP-ribosylating factor that is capable of halting protein synthesis in eukaryotic cells. The prevalence of the gene encoding cholix toxin, chxA, was assessed among a collection of 155 diverse V. cholerae strains originating from both clinical and environmental settings in Bangladesh and Mexico and other countries around the globe. The chxA gene was present in 47% of 83 non-O1, non-O139 strains and 16% of 72 O1/O139 strains screened as part of this study. A total of 86 chxA gene sequences were obtained, and phylogenetic analysis revealed that they fall into two distinct clades. These two clades were also observed in the phylogenies of several housekeeping genes, suggesting that the divergence observed in chxA extends to other regions of the V. cholerae genome, and most likely has arisen from vertical descent rather than horizontal transfer. Our results clearly indicate that ChxA is a major toxin of V. cholerae with a worldwide distribution that is preferentially associated with non-pandemic strains. © 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.

  3. Cholera toxin B subunit-binding and ganglioside GM1 immuno-expression are not necessarily correlated in human salivary glands

    DEFF Research Database (Denmark)

    Kirkeby, Svend

    2014-01-01

    human submandibular, parotid and palatinal glands using cholera toxin sub-unit B and two polyclonal antibodies against ganglioside GM1 as biomarkers. RESULTS: Immunofluorescence microscopy showed that the toxin and antibodies were co-localized in some acini but not in others. The cholera toxin mainly...... reacted with the cell membranes of the mucous acini in the submandibular gland, while incubation with the antibody against GM1 gave rise to a staining of the cytoplasm. The cytoplasm in some secretory acinar cells in the parotid gland was stained by the cholera toxin, whereas only small spots...... on the plasma membranes reacted with anti-GM1. The plasma membranes in the parotid excretory ducts appeared to react to anti-GM1, but not to cholera toxin. CONCLUSIONS: Cholera toxin induces the expression of ion channels and carriers in the small intestine and increases the production of secretory mucins...

  4. Fucosylation and protein glycosylation create functional receptors for cholera toxin

    DEFF Research Database (Denmark)

    Wands, Amberlyn M; Fujita, Akiko; McCombs, Janet E

    2015-01-01

    Cholera toxin (CT) enters and intoxicates host cells after binding cell surface receptors using its B subunit (CTB). The ganglioside (glycolipid) GM1 is thought to be the sole CT receptor; however, the mechanism by which CTB binding to GM1 mediates internalization of CT remains enigmatic. Here we...... in normal human intestinal epithelia and could play a role in cholera....

  5. Persistence of plasmids, cholera toxin genes, and prophage DNA in classical Vibrio cholerae O1.

    OpenAIRE

    Cook, W L; Wachsmuth, K; Johnson, S R; Birkness, K A; Samadi, A R

    1984-01-01

    Plasmid profiles, the location of cholera toxin subunit A genes, and the presence of the defective VcA1 prophage genome in classical Vibrio cholerae isolated from patients in Bangladesh in 1982 were compared with those in older classical strains isolated during the sixth pandemic and with those in selected eltor and nontoxigenic O1 isolates. Classical strains typically had two plasmids (21 and 3 megadaltons), eltor strains typically had no plasmids, and nontoxigenic O1 strains had zero to thr...

  6. 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. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. [The immunology of cholera and the molecular biology of cholera toxin. Recent progress and future perspectives].

    Science.gov (United States)

    Carrada-Bravo, T

    1994-01-01

    Vibrio cholerae has recently called the attention of researchers due to its strong immunogenicity and also because it serves as coadjunct immunomodulator of the immune response of the intestinal mucosae for the mixed added antigens as well as for those covalently linked to the toxin. The immunopathogeny of cholera is a complex phenomenon. This article presents the preliminary results of experiments conducted with laboratory rats in order to find the IgA intestinal response of rodents and humans.

  8. 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.

  9. 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.

  10. Action of cholera toxin in the intestinal epithelial cells

    International Nuclear Information System (INIS)

    Hyun, C.S.

    1982-01-01

    The primary event in the action of cholera toxin on the isolated chick intestinal epithelial cell is its interaction with the cell membrane. This involves a large number (17 million per cell) of high affinity binding sites which belong to a single class. Binding of biologically active 125 I-labeled toxin is rapid, temperature-dependent, reversible, and saturable over a wide range of concentrations and includes only a small contribution from nonspecific sites. A characteristic lag phase of 10 min occurs following the complete binding of toxin before any increase in cellular cAMP levels can be detected in the isolated cells. The response (elevation of cellular cAMP) of the enterocytes to cholera toxin is linear with time for 40-50 min and causes a six- to eight-fold increase over control levels at steady stae. cAMP and agents that increase cAMP production inhibit Cl - -independent Na + influx into the isolated enterocytes whereas chlorporomazine (CPZ) which completely abolishes toxin-induced elevation of cAMP both reverses and prevents the cAMP-mediated inhibition of Na + entry. Correlation between cellular cAMP levels and the magnitude of Na + influx into the enterocytes provides evidence for a cAMP-mediated control of intestinal Na + uptake, which may represent the mechanistic basis for the antiabsorptive effect of CT and Na + during induction of intestinal secretion. The effect of cAMP on Na + but no Cl - influx in our villus cell preparation can be partially explained in terms of a cAMP-regulated Na + /H + neutral exchange system

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

    International Nuclear Information System (INIS)

    Smirnova, N.I.; Livanova, L.F.; Shaginyan, I.A.; Motin, V.L.

    1986-01-01

    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

  12. Zinc oxide nanoparticles provide anti-cholera activity by disrupting the interaction of cholera toxin with the human GM1 receptor.

    Science.gov (United States)

    Sarwar, Shamila; Ali, Asif; Pal, Mahadeb; Chakrabarti, Pinak

    2017-11-03

    Vibrio cholerae causes cholera and is the leading cause of diarrhea in developing countries, highlighting the need for the development of new treatment strategies to combat this disease agent. While exploring the possibility of using zinc oxide (ZnO) nanoparticles (NPs) in cholera treatment, we previously found that ZnO NPs reduce fluid accumulation in mouse ileum induced by the cholera toxin (CT) protein. To uncover the mechanism of action of ZnO NPs on CT activity, here we used classical (O395) and El Tor (C6706) V. cholerae biotypes in growth and biochemical assays. We found that a ZnO NP concentration of 10 μg/ml did not affect the growth rates of these two strains, nor did we observe that ZnO NPs reduce the expression levels of CT mRNA and protein. It was observed that ZnO NPs form a complex with CT, appear to disrupt the CT secondary structure, and block its interaction with the GM1 ganglioside receptor in the outer leaflet of the plasma membrane in intestinal (HT-29) cells and thereby reduce CT uptake into the cells. In the range of 2.5-10 μg/ml, ZnO NPs exhibited no cytotoxicity on kidney (HEK293) and HT-29 cells. We conclude that ZnO NPs prevent the first step in the translocation of cholera toxin into intestinal epithelial cells without exerting measurable toxic effects on HEK293 and HT-29 cells. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Detection of Cholera Toxin by Optical Methods: A Mechanism-Based Approach to the Generic Detection of Protein Toxins

    National Research Council Canada - National Science Library

    Young, Ronald

    1997-01-01

    ... after its ADP-ribosylation. The sensitive technique of fluorescence spectroscopy can be employed to monitor the action of cholera toxin without regards to the substituents on the acceptor molecule by use of epsilon NAD...

  14. The 2.3 {angstrom} crystal structure of cholera toxin B subunit pentamer: Choleragenoid

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Rong-Guang; Westbrook, M.L. [Argonne National Lab., IL (United States); Maulik, P.R.; Reed, R.A.; Shipley, G. [Boston Univ., MA (United States). School of Medicine; Westbrook, E.M. [Argonne National Lab., IL (United States)]|[Northwestern Univ., Evanston, IL (United States); Scott, D.L.; Otwinowski, Z. [Yale Univ., New Haven, CT (United States)

    1996-02-01

    Cholera toxin, a heterohexameric AB{sub 5} enterotoxin released by Vibrio cholera, induces a profuse secretory diarrhea in susceptible hosts. Choleragenoid, the B subunit pentamer of cholera toxin, directs the enzymatic A subunit to its target by binding to GM{sub 1} gangliosides exposed on the luminal surface of intestinal epithelial cells. We have solved the crystal structure of choleragenoid at 2.3 {Angstrom} resolution by combining single isomorphous replacement with non-crystallographic symmetry averaging. The structure of the B subunits, and their pentameric arrangement, closely resembles that reported for the intact holotoxin (choleragen), the heat-labile enterotoxin from E. coli, and for a choleragenoid-GM{sub 1} pentasaccharide complex. In the absence of the A subunit the central cavity of the B pentamer is a highly solvated channel. The binding of the A subunit or the receptor pentasaccharide to choleragenoid has only a modest effect on the local stereochemistry and does not perceptibly alter the subunit interface.

  15. Retrograde and transganglionic transport of horseradish peroxidase-conjugated cholera toxin B subunit, wheatgerm agglutinin and isolectin B4 from Griffonia simplicifolia I in primary afferent neurons innervating the rat urinary bladder.

    Science.gov (United States)

    Wang, H F; Shortland, P; Park, M J; Grant, G

    1998-11-01

    myelinated fibres). Double labelling with other neuronal markers showed that 71%, 43% and 36% of the cholera toxin B subunit-immunoreactive cells were calcitonin gene-related peptide-, isolectin B4-binding- and substance P-positive, respectively. A few cholera toxin B subunit cells showed galanin-immunoreactivity, but none were somatostatin-, vasoactive intestinal polypeptide-, or neuropeptide Y-immunoreactive or contained fluoride-resistant acid phosphatase. The results show that cholera toxin B subunit-horseradish peroxidase is a more effective retrograde and transganglionic tracer for pelvic primary afferents from the urinary bladder than wheat germ agglutinin-horseradish peroxidase and isolectin B4-horseradish peroxidase, but in contrast to somatic nerves, it is transported mainly by unmyelinated fibres in the visceral afferents.

  16. Role of the RS1 sequence of the cholera vibrio in amplification of the segment of plasmid DNA carrying the gene of resistance to tetracycline and the genes of cholera toxin

    International Nuclear Information System (INIS)

    Fil'kova, S.L.; Il'ina, T.S.; Gintsburg, A.L.; Yanishevskii, N.V.; Smirnov, G.B.

    1988-01-01

    The hybrid plasmid pCO107, representing cointegrate 14(2)-5(2) of two plasmids, an F-derivative (pOX38) and a PBR322-derivative (pCT105) with an RS1 sequence of the cholera vibrio cloned in its makeup, contains two copes of RS1 at the sites of union of the two plasmids. Using a tetracycline resistance marker (Tc R ) of the plasmid pCT105, clones were isolated which have an elevated level of resistance to tetracycline (an increase of from 4- to 30-fold). Using restriction analysis and the Southern blot method of hybridization it was shown that the increase in the level of resistance of tetracycline is associated with the amplification of pCT105 portion of the cointegrate, and that the process of amplification is governed by the presence of direct repeats of the RS1 sequence at its ends. The increase in the number of copies of the pCT105 segment, which contains in its composition the genes of cholera toxin (vct), is accompanied by an increase in toxin production

  17. Pilot study of whole-blood gamma interferon response to the Vibrio cholerae toxin B subunit and resistance to enterotoxigenic Escherichia coli-associated diarrhea.

    Science.gov (United States)

    Flores, Jose; DuPont, Herbert L; Paredes-Paredes, Mercedes; Aguirre-Garcia, M Magdalena; Rojas, Araceli; Gonzalez, Alexei; Okhuysen, Pablo C

    2010-05-01

    Enterotoxigenic Escherichia coli (ETEC), which produces heat-labile toxin (LT), is a common cause of travelers' diarrhea (TD). The B subunit of ETEC LT is immunologically related to the B subunit of Vibrio cholerae toxin (CT). In this pilot study we evaluated the whole-blood gamma interferon response to CT B in 17 U.S. adults traveling to Mexico. Only one of nine subjects who demonstrated a cellular immune response as determined by whole-blood gamma interferon production to CT B on arrival to Mexico developed diarrhea, whereas five of eight without a cellular response developed diarrhea. Markers of the cellular immune response to ETEC LT could help in identifying individuals immune to ETEC LT, and these markers deserve additional study.

  18. Virulence and the Environment: a Novel Role for Vibrio cholerae Toxin-Coregulated Pili in Biofilm Formation on Chitin

    Science.gov (United States)

    Reguera, Gemma; Kolter, Roberto

    2005-01-01

    The toxin-coregulated pilus (TCP) of Vibrio cholerae is required for intestinal colonization and cholera toxin acquisition. Here we report that TCP mediates bacterial interactions required for biofilm differentiation on chitinaceous surfaces. We also show that undifferentiated TCP− biofilms have reduced ecological fitness and, thus, that chitin colonization may represent an ecological setting outside the host in which selection for a host colonization factor may take place. PMID:15866944

  19. 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

    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...... in the fimA gene. This was carried out by introduction of new restriction sites by PCR-mediated site-directed mutagenesis of fimA in positions predicted to correspond to optimally surface-located regions of the subunit protein. Subsequently, the synthetic cholera-toxin-encoding DNA segment was inserted....... 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...

  20. The inhibition of cholera toxin-induced 5-HT release by the 5-HT3 receptor antagonist, granisetron, in the rat

    Science.gov (United States)

    Turvill, J L; Connor, P; Farthing, M J G

    2000-01-01

    The secretagogue 5-hydroxytryptamine (5-HT) is implicated in the pathophysiology of cholera. 5-HT released from enterochromaffin cells after cholera toxin exposure is thought to activate non-neuronally (5-HT2 dependent) and neuronally (5-HT3 dependent) mediated water and electrolyte secretion. CT-secretion can be reduced by preventing the release of 5-HT. Enterochromaffin cells possess numerous receptors that, under basal conditions, modulate 5-HT release. These include basolateral 5-HT3 receptors, the activation of which is known to enhance 5-HT release. Until now, 5-HT3 receptor antagonists (e.g. granisetron) have been thought to inhibit cholera toxin-induced fluid secretion by blockading 5-HT3 receptors on secretory enteric neurones. Instead we postulated that they act by inhibiting cholera toxin-induced enterochromaffin cell degranulation. Isolated intestinal segments in anaesthetized male Wistar rats, pre-treated with granisetron 75 μg kg−1, lidoocaine 6 mg kg−1 or saline, were instilled with a supramaximal dose of cholera toxin or saline. Net fluid movement was determined by small intestinal perfusion or gravimetry and small intestinal and luminal fluid 5-HT levels were determined by HPLC with fluorimetric detection. Intraluminal 5-HT release was proportional to the reduction in tissue 5-HT levels and to the onset of water and electrolyte secretion, suggesting that luminal 5-HT levels reflect enterochromaffin cell activity. Both lidocaine and granisetron inhibited fluid secretion. However, granisetron alone, and proportionately, reduced 5-HT release. The simultaneous inhibition of 5-HT release and fluid secretion by granisetron suggests that 5-HT release from enterochromaffin cells is potentiated by endogenous 5-HT3 receptors. The accentuated 5-HT release promotes cholera toxin-induced fluid secretion. PMID:10882387

  1. Cholera

    Science.gov (United States)

    ... causes a large amount of watery diarrhea. Causes Cholera is caused by the bacterium Vibrio cholerae . These bacteria release a toxin that causes an ... include: Africa Some parts of Asia India Bangladesh Mexico South and Central America ... Symptoms of cholera can be mild to severe. They include: Abdominal ...

  2. Plasma and Mucosal Immunoglobulin M, Immunoglobulin A, and Immunoglobulin G Responses to the Vibrio cholerae O1 Protein Immunome in Adults With Cholera in Bangladesh.

    Science.gov (United States)

    Charles, Richelle C; Nakajima, Rie; Liang, Li; Jasinskas, Al; Berger, Amanda; Leung, Daniel T; Kelly, Meagan; Xu, Peng; Kovác, Pavol; Giffen, Samantha R; Harbison, James D; Chowdhury, Fahima; Khan, Ashraful I; Calderwood, Stephen B; Bhuiyan, Taufiqur Rahman; Harris, Jason B; Felgner, Philip L; Qadri, Firdausi; Ryan, Edward T

    2017-07-01

    Cholera is a severe dehydrating illness of humans caused by toxigenic strains of Vibrio cholerae O1 or O139. Identification of immunogenic V. cholerae antigens could lead to a better understanding of protective immunity in human cholera. We probed microarrays containing 3652 V. cholerae antigens with plasma and antibody-in-lymphocyte supernatant (ALS, a surrogate marker of mucosal immune responses) from patients with severe cholera caused by V. cholerae O1 in Bangladesh and age-, sex-, and ABO-matched Bangladeshi controls. We validated a subset of identified antigens using enzyme-linked immunosorbent assay. Overall, we identified 608 immunoreactive V. cholerae antigens in our screening, 59 of which had higher immunoreactivity in convalescent compared with acute-stage or healthy control samples (34 in plasma, 39 in mucosal ALS; 13 in both sample sets). Identified antigens included cholera toxin B and A subunits, V. cholerae O-specific polysaccharide and lipopolysaccharide, toxin coregulated pilus A, sialidase, hemolysin A, flagellins (FlaB, FlaC, and FlaD), phosphoenolpyruvate-protein phosphotransferase, and diaminobutyrate-2-oxoglutarate aminotransferase. This study is the first antibody profiling of the mucosal and systemic antibody responses to the nearly complete V. cholerae O1 protein immunome; it has identified antigens that may aid in the development of an improved cholera vaccine. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America.

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

    Science.gov (United States)

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

    2014-01-01

    Vibrio cholerae O1 classical, El Tor and O139 are the primary biotypes that cause epidemic cholera, and they also express cholera toxin (CT). Although classical V. cholerae produces CT in various settings, the El Tor and O139 strains require specific growth conditions for CT induction, such as the so-called AKI conditions, which consist of growth in static conditions followed by growth under aerobic shaking conditions. However, our group has demonstrated that CT production may also take place in shallow static cultures. How these type of cultures induce CT production has been unclear, but we now report that in shallow culture growth conditions, there is virtual depletion of dissolved oxygen after 2.5 h of growth. Concurrently, during the first three to 4 h, endogenous CO2 accumulates in the media and the pH decreases. These findings may explain CT expression at the molecular level because CT production relies on a regulatory cascade, in which the key regulator AphB may be activated by anaerobiosis and by low pH. AphB activation stimulates TcpP synthesis, which induces ToxT production, and ToxT directly stimulates ctxAB expression, which encodes CT. Importantly, ToxT activity is enhanced by bicarbonate. Therefore, we suggest that in shallow cultures, AphB is activated by initial decreases in oxygen and pH, and subsequently, ToxT is activated by intracellular bicarbonate that has been generated from endogenous CO2. This working model would explain CT production in shallow cultures and, possibly, also in other growth conditions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Cholera toxin B subunit labeling in lamina II of spinal cord dorsal horn following chronic inflammation in rats.

    Science.gov (United States)

    Ma, Qing Ping; Tian, Li

    2002-07-26

    We have investigated the effect of inflammation on the labeling pattern of cholera toxin B subunit (CTB)-conjugated horseradish peroxidase, an A-fiber marker, by an intra-sciatic nerve injection of the tracer. Following chronic inflammation in one hind paw in rats, there was substantial CTB labeling in lamina II of the spinal dorsal horn, which is normally absent. However, there was no change in the labeling pattern of wheat germ agglutinin or fluoride resistant acid phosphatase/thiamine monophosphatase, two C-fiber markers. The CTB labeling in lamina II after peripheral nerve injury has been interpreted as central sprouting of A-fibers or uptake of the tracer by injured C-fibers. Our results suggest that chronic inflammation and nerve injury may share some common mechanisms in generating allodynia and hyperalgesia.

  5. Crystallization of the HigBA2 toxin-antitoxin complex from Vibrio cholerae

    DEFF Research Database (Denmark)

    Hadǽi, San; Garcia-Pino, Abel; Martinez-Rodriguez, Sergio

    2013-01-01

    The genome of Vibrio cholerae encodes two higBA toxin-antitoxin (TA) modules that are activated by amino-acid starvation. Here, the TA complex of the second module, higBA2, as well as the C-terminal domain of the corresponding HigA2 antitoxin, have been purified and crystallized. The HigBA2 compl...

  6. Biomarkers of Environmental Enteropathy are Positively Associated with Immune Responses to an Oral Cholera Vaccine in Bangladeshi Children.

    Directory of Open Access Journals (Sweden)

    Muhammad Ikhtear Uddin

    2016-11-01

    Full Text Available Environmental enteropathy (EE is a poorly understood condition that refers to chronic alterations in intestinal permeability, absorption, and inflammation, which mainly affects young children in resource-limited settings. Recently, EE has been linked to suboptimal oral vaccine responses in children, although immunological mechanisms are poorly defined. The objective of this study was to determine host factors associated with immune responses to an oral cholera vaccine (OCV. We measured antibody and memory T cell immune responses to cholera antigens, micronutrient markers in blood, and EE markers in blood and stool from 40 Bangladeshi children aged 3-14 years who received two doses of OCV given 14 days apart. EE markers included stool myeloperoxidase (MPO and alpha anti-trypsin (AAT, and plasma endotoxin core antibody (EndoCab, intestinal fatty acid binding protein (i-FABP, and soluble CD14 (sCD14. We used multiple linear regression analysis with LASSO regularization to identify host factors, including EE markers, micronutrient (nutritional status, age, and HAZ score, predictive for each response of interest. We found stool MPO to be positively associated with IgG antibody responses to the B subunit of cholera toxin (P = 0.03 and IgA responses to LPS (P = 0.02; plasma sCD14 to be positively associated with LPS IgG responses (P = 0.07; plasma i-FABP to be positively associated with LPS IgG responses (P = 0.01 and with memory T cell responses specific to cholera toxin (P = 0.01; stool AAT to be negatively associated with IL-10 (regulatory T cell responses specific to cholera toxin (P = 0.02, and plasma EndoCab to be negatively associated with cholera toxin-specific memory T cell responses (P = 0.02. In summary, in a cohort of children 3-14 years old, we demonstrated that the majority of biomarkers of environmental enteropathy were positively associated with immune responses after vaccination with an OCV.

  7. Purification and characterization of Yersinia enterocolitica and Yersinia pestis LcrV-cholera toxin A(2)/B chimeras.

    Science.gov (United States)

    Tinker, Juliette K; Davis, Chadwick T; Arlian, Britni M

    2010-11-01

    Yersinia pestis is a virulent human pathogen and potential biological weapon. Despite a long history of research on this organism, there is no licensed vaccine to protect against pneumonic forms of Y. pestis disease. In the present study, plasmids were constructed to express cholera toxin A(2)/B chimeric molecules containing the LcrV protective antigen from Yersinia enterocolitica and Y. pestis. These chimeras were expressed and purified to high yields from the supernatant of transformed Escherichia coli. Western and GM(1) ELISA assays were used to characterize the composition, receptor-binding and relative stability of the LcrV-CTA(2)/B chimera in comparison to cholera toxin. In addition, we investigated the ability of the Y. pestis LcrV-CTA(2)/B chimera to bind to and internalize into cultured epithelial cells and macrophages by confocal microscopy. These studies indicate that the uptake and trafficking of the LcrV antigen from the chimera is comparable to the trafficking of native toxin. Together these findings report that stable, receptor-binding, non-toxic LcrV-cholera toxin A(2)/B chimeras can be expressed at high levels in E. coli and purified from the supernatant. In addition, the internalization of antigen in vitro reported here supports the development of these molecules as novel mucosal vaccine candidates. Copyright 2010 Elsevier Inc. All rights reserved.

  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.

  9. Binding of fluorescently labeled cholera toxin subunit B to glycolipids in the human submandibular gland and inhibition of binding by periodate oxidation and by galactose

    DEFF Research Database (Denmark)

    Kirkeby, S

    2016-01-01

    FITC-labeled cholera toxin subunit B (CTB) stained the surfaces of cells of mucous acini in the submandibular gland. CTB, also called choleragenoid, binds to the GM1 glycolipid in the cell membrane. The binding in most acini was inhibited by periodic acid oxidation of the sections, while some acini...... to the internal galactose residue linked to GalNAc, as in the GM1 glycolipid. Inhibition of the GM1 receptor binding to cholera toxin has potential for protection of humans against cholera. Galactose and agents that modify sialic acid inhibit the accessibility of the toxin to the GM1 carbohydrate receptor. Human...

  10. Cholera toxin-induced ADP-ribosylation of a 46 kDa protein is decreased in brains of ethanol-fed mice

    International Nuclear Information System (INIS)

    Nhamburo, P.T.; Hoffman, P.L.; Tabakoff, B.

    1988-01-01

    The acute in vitro effects of ethanol on cerebral cortical adenylate cyclase activity and beta-adrenergic receptor characteristics suggested a site of action of ethanol at Gs, the stimulatory guanine nucleotide binding protein. After chronic ethanol ingestion, the beta-adrenergic receptor appeared to be uncoupled (i.e., the form of the receptor with high affinity for agonist was undetectable), and stimulation of adenylate cyclase activity by isoproterenol or guanine nucleotides was reduced, suggesting an alteration in the properties of Gs. To further characterize this change, cholera and pertussis toxin-mediated 32 P-ADP-ribosylation of mouse cortical membranes was assessed in mice that had chronically ingested ethanol in a liquid diet. 32 P-labeled proteins were separated by SDS-PAGE and quantitated by autoradiography. There was a selective 30-50% decrease in cholera toxin-induced labeling of 46 kDa protein band in membranes of ethanol-fed mice, with no apparent change in pertussis toxin-induced labeling. The 46 kDa protein has a molecular weight similar to that of the alpha subunit of Gs, suggesting a reduced amount of this protein or a change in its characteristics as a substrate for cholera toxin-induced ADP-ribosylation in cortical membranes of ethanol-fed mice

  11. Live Staining and Isolation of Specific Hormone-Producing Cells from Rat Anterior Pituitary by Cytochemistry with Lectins and Cholera Toxin B Subunit

    International Nuclear Information System (INIS)

    Kikuchi, Motoshi; Kusumoto, Kenji; Fujiwara, Ken; Takahashi, Kozue; Tando, Yukiko; Yashiro, Takashi

    2011-01-01

    Anterior pituitary glands contain five types of hormone-producing cells. Distinguishing and isolating specific types of living cells are essential for studying their function. Although many such attempts have been made, the results have been disappointing. In the present study, we labeled specific types of living hormone-producing cells by using potential differences in sugar chains on the cell surfaces. Cytochemical analysis with lectins and cholera toxin B subunit revealed that PNA, S-WGA, and cholera toxin B subunit recognized sugar chains specific to prolactin cells, ACTH cells, and GH cells, respectively, and that UEA-I recognized most of prolactin cells and GH cells. Next, fluorescence-activated cell sorting was used to isolate GH cells labeled by fluoresceinated cholera toxin B. The purity of the GH cell fraction estimated by immunocytochemistry and quantitative real-time PCR for cell type-specific genes was more than 98%, which was higher than that reported in earlier studies, including those using transgenic animals. We conclude that cytochemistry with lectins and cholera toxin B subunit is a straightforward, acceptable method of isolating specific types of anterior pituitary cells and that the cells isolated by this method can serve as useful materials in the study of anterior pituitary cells

  12. Recombinant cholera toxin B subunit in Escherichia coli: high-level secretion, purification, and characterization

    NARCIS (Netherlands)

    Slos, P.; Speck, D.; Accart, N.; Kolbe, H.V.; Schubnel, D.; Bouchon, B.; Bischoff, Rainer; Kieny, M.P.

    1994-01-01

    The gene coding for cholera toxin subunit B (CT-B) was fused to a modified ompA signal sequence and subsequently cloned into a high expression vector based on the regulatory signals of the arabinose operon of Salmonella typhimurium. Upon induction of gene expression in Escherichia coli, a product of

  13. Cholera toxin B subunits assemble into pentamers--proposition of a fly-casting mechanism.

    Directory of Open Access Journals (Sweden)

    Jihad Zrimi

    Full Text Available The cholera toxin B pentamer (CtxB(5, which belongs to the AB(5 toxin family, is used as a model study for protein assembly. The effect of the pH on the reassembly of the toxin was investigated using immunochemical, electrophoretic and spectroscopic methods. Three pH-dependent steps were identified during the toxin reassembly: (i acquisition of a fully assembly-competent fold by the CtxB monomer, (ii association of CtxB monomer into oligomers, (iii acquisition of the native fold by the CtxB pentamer. The results show that CtxB(5 and the related heat labile enterotoxin LTB(5 have distinct mechanisms of assembly despite sharing high sequence identity (84% and almost identical atomic structures. The difference can be pinpointed to four histidines which are spread along the protein sequence and may act together. Thus, most of the toxin B amino acids appear negligible for the assembly, raising the possibility that assembly is driven by a small network of amino acids instead of involving all of them.

  14. Inhibition of Binding of the AB5-Type Enterotoxins LT-I and Cholera Toxin to Ganglioside GM1 by Galactose-Rich Dietary Components

    NARCIS (Netherlands)

    Becker, P.M.; Widjaja-Greefkes, H.C.A.; Wikselaar, van P.G.

    2010-01-01

    Cholera, travelers' diarrhea, or colibacillosis in pigs can possibly be prevented or attenuated by dietary provision of competitive inhibitors that react with the GM1-binding sites of the enterotoxins cholera toxin (CT), human Escherichia coli heat-labile enterotoxin of serogroup I (LTh-I), and

  15. NADP+ enhances cholera and pertussis toxin-catalyzed ADP-ribosylation of membrane proteins

    International Nuclear Information System (INIS)

    Kawai, Y.; Whitsel, C.; Arinze, I.J.

    1986-01-01

    Cholera or pertussis toxin-catalyzed [ 32 P]ADP-ribosylation is frequently used to estimate the concentration of the stimulatory (Ns) or inhibitory (Ni) guanine nucleotide regulatory proteins which modulate the activity of adenylate cyclase. With this assay, however, the degradation of the substrate, NAD + , by endogenous enzymes such as NAD + -glycohydrolase (NADase) present in the test membranes can influence the results. In this study the authors show that both cholera and pertussis toxin-catalyzed [ 32 P]ADP-ribosylation of liver membrane proteins is markedly enhanced by NADP + . The effect is concentration dependent; with 20 μM [ 32 P]NAD + as substrate maximal enhancement is obtained at 0.5-1.0 mM NADP + . The enhancement of [ 32 P]ADP-ribosylation by NADP + was much greater than that by other known effectors such as Mg 2+ , phosphate or isoniazid. The effect of NADP + on ADP-ribosylation may occur by inhibition of the degradation of NAD + probably by acting as an alternate substrate for NADase. Among inhibitors tested (NADP + , isoniazid, imidazole, nicotinamide, L-Arg-methyl-ester and HgCl 2 ) to suppress NADase activity, NADP + was the most effective and, 10 mM, inhibited activity of the enzyme by about 90%. In membranes which contain substantial activities of NADase the inclusion of NADP + in the assay is necessary to obtain maximal ADP-ribosylation

  16. Platelet cytosolic 44-kDa protein is a substrate of cholera toxin-induced ADP-ribosylation and is not recognized by antisera against the α subunit of the stimulatory guanine nucleotide-binding regulatory protein

    International Nuclear Information System (INIS)

    Molina Y Vedia, L.M.; Reep, B.R.; Lapetina, E.G.

    1988-01-01

    ADP-ribosylation induced by cholera toxin and pertussis toxin was studied in particulate and cytosolic fractions of human platelets. Platelets were disrupted by a cycle of freezing and thawing in the presence of a hyposmotic buffer containing protease inhibitors. In both fractions, the A subunit of cholera toxin ADP-ribosylates two proteins with molecular masses of 42 and 44 kDa, whereas pertussis toxin ADP-ribosylates a 41-kDa polypeptide. Two antisera against the α subunit of the stimulatory guanine nucleotide-binding regulatory protein recognize only the 42-kDa polypeptide. Cholera toxin-induced ADP-ribosylation of the 42- and 44-kDa proteins is reduced by pretreatment of platelets with iloprost, a prostacyclin analog. The 44-kDa protein, which is substrate of cholera toxin, could be extracted completely from the membrane and recovered in the cytosolic fraction when the cells were disrupted by Dounce homogenization and the pellet was extensively washed. A 44-kDa protein can also be labeled with 8-azidoguanosine 5'-[α- 32 P]triphosphate in the cytosol and membranes. These finding indicate that cholera and pertussis toxins produced covalent modifications of proteins present in particulate and cytosolic platelet fractions. Moreover, the 44-kDa protein might be an α subunit of a guanine nucleotide-binding regulatory protein that is not recognized by available antisera

  17. [Cytotoxic effect of Vibrio cholerae non-O1 on Vero cells].

    Science.gov (United States)

    Figueroa-Arredondo, P; García-Lozano, H; Gutiérrez-Cogco, L; Valdespino-Gómez, J L

    1994-01-01

    At the present time there is still in Mexico a diarrhoeal outbreak due to Vibrio cholerae O1. In INDRE we have isolated from the same outbreak last year (jan-apr), 70 strains of Vibrio cholerae Non-O1. These were isolated from patients with a diarrhoeal illness different from cholera. Patients were of different ages and sex, and from various geographic areas. The isolated strains were confirmed by serological agglutination test with polyclonal antisera, and they neither belong to O1 serogroup or O139. We assayed all the 70 strains in Vero cells, searching for cytotoxic effect, probably attributed to cholera toxin, or any other toxin. The strains were screened by PCR for cholera toxin gene detection, and negative results were obtained. We have found only one CT-producer strain, but it was a rough one so, we are not able to affirm that is not a V. cholerae O1 serotype. Vibrio cholerae Non-O1 strains, tested in Vero cells assay, produced cytotoxic effect within 24 h. It was found that 48/70 strains (66.6%), had cytotoxic activity, showing rounding and then lysis of cells. From our results we concluded that this cytotoxic effect, is not cholera toxin related, instead we propose it could be due to an unknown virulence factor, probably a different toxin in mexican Vibrio cholerae Non-O1 strains.

  18. Therapeutic Potential of Cholera Toxin B Subunit for the Treatment of Inflammatory Diseases of the Mucosa

    Directory of Open Access Journals (Sweden)

    Joshua M. Royal

    2017-11-01

    Full Text Available Cholera toxin B subunit (CTB is a mucosal immunomodulatory protein that induces robust mucosal and systemic antibody responses. This well-known biological activity has been exploited in cholera prevention (as a component of Dukoral® vaccine and vaccine development for decades. On the other hand, several studies have investigated CTB’s immunotherapeutic potential in the treatment of inflammatory diseases such as Crohn’s disease and asthma. Furthermore, we recently found that a variant of CTB could induce colon epithelial wound healing in mouse colitis models. This review summarizes the possible mechanisms behind CTB’s anti-inflammatory activity and discuss how the protein could impact mucosal inflammatory disease treatment.

  19. The population structure of Vibrio cholerae from the Chandigarh Region of Northern India.

    Directory of Open Access Journals (Sweden)

    Moataz Abd El Ghany

    2014-07-01

    Full Text Available Cholera infection continues to be a threat to global public health. The current cholera pandemic associated with Vibrio cholerae El Tor has now been ongoing for over half a century.Thirty-eight V. cholerae El Tor isolates associated with a cholera outbreak in 2009 from the Chandigarh region of India were characterised by a combination of microbiology, molecular typing and whole-genome sequencing. The genomic analysis indicated that two clones of V. cholera circulated in the region and caused disease during this time. These clones fell into two distinct sub-clades that map independently onto wave 3 of the phylogenetic tree of seventh pandemic V. cholerae El Tor. Sequence analyses of the cholera toxin gene, the Vibrio seventh Pandemic Island II (VSPII and SXT element correlated with this phylogenetic position of the two clades on the El Tor tree. The clade 2 isolates, characterized by a drug-resistant profile and the expression of a distinct cholera toxin, are closely related to the recent V. cholerae isolated elsewhere, including Haiti, but fell on a distinct branch of the tree, showing they were independent outbreaks. Multi-Locus Sequence Typing (MLST distinguishes two sequence types among the 38 isolates, that did not correspond to the clades defined by whole-genome sequencing. Multi-Locus Variable-length tandem-nucleotide repeat Analysis (MLVA identified 16 distinct clusters.The use of whole-genome sequencing enabled the identification of two clones of V. cholerae that circulated during the 2009 Chandigarh outbreak. These clones harboured a similar structure of ICEVchHai1 but differed mainly in the structure of CTX phage and VSPII. The limited capacity of MLST and MLVA to discriminate between the clones that circulated in the 2009 Chandigarh outbreak highlights the value of whole-genome sequencing as a route to the identification of further genetic markers to subtype V. cholerae isolates.

  20. Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin

    DEFF Research Database (Denmark)

    Rissanen, Sami; Grzybek, Michal; Orłowski, Adam

    2017-01-01

    membrane vesicles and giant unilamellar vesicles, specific binding of Cholera Toxin (CTxB) to GM1 glycolipids is a commonly used strategy to label raft domains or Lo membrane environments. However, these studies often use acyl-chain labeled bodipy-GM1 (bdGM1), whose headgroup accessibility and membrane...

  1. Authentic display of a cholera toxin epitope by chimeric type 1 fimbriae

    DEFF Research Database (Denmark)

    Stentebjerg-Olesen, Bodil; Pallesen, Lars; Jensen, Lars Bogø

    1997-01-01

    . 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...... 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...

  2. The protective activity of tea against infection by Vibrio cholerae O1.

    Science.gov (United States)

    Toda, M; Okubo, S; Ikigai, H; Suzuki, T; Suzuki, Y; Shimamura, T

    1991-02-01

    Extracts of black tea exhibited bactericidal activity against Vibrio cholerae O1. The tea extract inhibited the haemolysin activity of V. cholerae O1, El Tor and the morphological changes of Chinese hamster ovary cells induced by cholera toxin. Tea extract also reduced fluid accumulation induced by cholera toxin in sealed adult mice and by V. cholerae O1 in ligated intestinal loops of rabbits. These findings suggest that tea has protective activity against V. cholerae O1.

  3. Action of cholera toxin in the intestinal epithelial cells

    International Nuclear Information System (INIS)

    Hyun, C.S.

    1982-01-01

    The primary event in the action of cholera toxin on the isolated chick intestinal epithelial cell is its interaction with a large number of high affinity binding sites in the cell membrane. Binding of 125 I-labeled toxin is rapid, temperature-dependent, reversible, and saturable over a wide range of concentrations and includes only a small contribution from nonspecific sites. A characteristic lag phase of 10 min occurs following the complete binding of toxin before any increase in cellular cAMP levels can be detected. The response (elevation of cellular cAMP) is linear with time for 40 to 50 min and causes a six- to eight-fold increase over control levels (10 to 15 picomole cAMP/mg cellular protein) at steady state. cAMP and agents that increase cAMP production inhibit Cl - -independent Na + influx into the isolated enterocytes whereas chlorpromazine (CPZ) which completely abolishes toxin-induced elevation of cAMP both reverses and prevents the cAMP-mediated inhibition of Na + entry. Correlation between cellular cAMP levels and the magnitude of Na + influx provides evidence for a cAMP-mediated control of intestinal Na + uptake, which may represent the mechanistic basis for the antiabsorptive effect of CT on Na + during induction of intestinal secretion. The effect of cAMP on Na + but not Cl - influx preparations can be partially explained in terms of a cAMP-regulated Na + /H + neutral exchange system. Data on the coupling relationship between Na + transport and the intra- and extracellular pH in the enterocytes show that an amiloride-sensitive electroneutral Na + /H + exchange process occurs. This coupling between Na + and H + is partially inhibited by CT and dbcAMP, suggesting that the Na + /H + exchange may be a cAMP-regulated process. 31 references, 32 figures, 5 tables

  4. High-Resolution FRET Microsopy of Cholera Toxin B-Subunit and GPI-anchored Proteins in Cell Plasma Mambranes

    Czech Academy of Sciences Publication Activity Database

    Kenworthy, A. K.; Brdičková, Naděžda; Edidin, M.

    2000-01-01

    Roč. 11, č. 5 (2000), s. 1645-1655 ISSN 0248-4900 R&D Projects: GA AV ČR IAA7052904 Institutional research plan: CEZ:AV0Z5052915 Keywords : High-Resolution * Cholera Toxin * Cell Plasma Membrane s Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.670, year: 2000

  5. Effect of in vivo injection of cholera and pertussis toxin on glucose transport in rat skeletal muscle

    DEFF Research Database (Denmark)

    Ploug, Thorkil; Han, X; Petersen, L N

    1997-01-01

    Cholera toxin (CTX) and pertussis toxin (PTX) were examined for their ability to inhibit glucose transport in perfused skeletal muscle. Twenty-five hours after an intravenous injection of CTX, basal transport was decreased approximately 30%, and insulin- and contraction-stimulated transport...... in GLUT-1 protein content was found. In contrast, GLUT-4 mRNA was unchanged, but transcripts for GLUT-1 were increased > or = 150% in all three muscles from CTX-treated rats. The findings suggest that CTX via increased cAMP impairs basal as well as insulin- and contraction-stimulated muscle glucose...

  6. Rotavirus 2/6 Viruslike Particles Administered Intranasally with Cholera Toxin, Escherichia coli Heat-Labile Toxin (LT), and LT-R192G Induce Protection from Rotavirus Challenge

    OpenAIRE

    O’Neal, Christine M.; Clements, John D.; Estes, Mary K.; Conner, Margaret E.

    1998-01-01

    We have shown that rotavirus 2/6 viruslike particles composed of proteins VP2 and VP6 (2/6-VLPs) administered to mice intranasally with cholera toxin (CT) induced protection from rotavirus challenge, as measured by virus shedding. Since it is unclear if CT will be approved for human use, we evaluated the adjuvanticity of Escherichia coli heat-labile toxin (LT) and LT-R192G. Mice were inoculated intranasally with 10 μg of 2/6-VLPs combined with CT, LT, or LT-R192G. All three adjuvants induced ...

  7. Intracellular Signal Triggered by Cholera Toxin in Saccharomyces boulardii and Saccharomyces cerevisiae

    Science.gov (United States)

    Brandão, Rogelio L.; Castro, Ieso M.; Bambirra, Eduardo A.; Amaral, Sheila C.; Fietto, Luciano G.; Tropia, Maria José M.; Neves, Maria José; Dos Santos, Raquel G.; Gomes, Newton C. M.; Nicoli, Jacques R.

    1998-01-01

    As is the case for Saccharomyces boulardii, Saccharomyces cerevisiae W303 protects Fisher rats against cholera toxin (CT). The addition of glucose or dinitrophenol to cells of S. boulardii grown on a nonfermentable carbon source activated trehalase in a manner similar to that observed for S. cerevisiae. The addition of CT to the same cells also resulted in trehalase activation. Experiments performed separately on the A and B subunits of CT showed that both are necessary for activation. Similarly, the addition of CT but not of its separate subunits led to a cyclic AMP (cAMP) signal in both S. boulardii and S. cerevisiae. These data suggest that trehalase stimulation by CT probably occurred through the cAMP-mediated protein phosphorylation cascade. The requirement of CT subunit B for both the cAMP signal and trehalase activation indicates the presence of a specific receptor on the yeasts able to bind to the toxin, a situation similar to that observed for mammalian cells. This hypothesis was reinforced by experiments with 125I-labeled CT showing specific binding of the toxin to yeast cells. The adhesion of CT to a receptor on the yeast surface through the B subunit and internalization of the A subunit (necessary for the cAMP signal and trehalase activation) could be one more mechanism explaining protection against the toxin observed for rats treated with yeasts. PMID:9464394

  8. The Population Structure of Vibrio cholerae from the Chandigarh Region of Northern India

    KAUST Repository

    Abd El Ghany, Moataz

    2014-07-24

    Background:Cholera infection continues to be a threat to global public health. The current cholera pandemic associated with Vibrio cholerae El Tor has now been ongoing for over half a century.Methodology/Principal Findings:Thirty-eight V. cholerae El Tor isolates associated with a cholera outbreak in 2009 from the Chandigarh region of India were characterised by a combination of microbiology, molecular typing and whole-genome sequencing. The genomic analysis indicated that two clones of V. cholera circulated in the region and caused disease during this time. These clones fell into two distinct sub-clades that map independently onto wave 3 of the phylogenetic tree of seventh pandemic V. cholerae El Tor. Sequence analyses of the cholera toxin gene, the Vibrio seventh Pandemic Island II (VSPII) and SXT element correlated with this phylogenetic position of the two clades on the El Tor tree. The clade 2 isolates, characterized by a drug-resistant profile and the expression of a distinct cholera toxin, are closely related to the recent V. cholerae isolated elsewhere, including Haiti, but fell on a distinct branch of the tree, showing they were independent outbreaks. Multi-Locus Sequence Typing (MLST) distinguishes two sequence types among the 38 isolates, that did not correspond to the clades defined by whole-genome sequencing. Multi-Locus Variable-length tandem-nucleotide repeat Analysis (MLVA) identified 16 distinct clusters.Conclusions/Significance:The use of whole-genome sequencing enabled the identification of two clones of V. cholerae that circulated during the 2009 Chandigarh outbreak. These clones harboured a similar structure of ICEVchHai1 but differed mainly in the structure of CTX phage and VSPII. The limited capacity of MLST and MLVA to discriminate between the clones that circulated in the 2009 Chandigarh outbreak highlights the value of whole-genome sequencing as a route to the identification of further genetic markers to subtype V. cholerae isolates.

  9. The Population Structure of Vibrio cholerae from the Chandigarh Region of Northern India

    KAUST Repository

    Abd El Ghany, Moataz; Chander, Jagadish; Mutreja, Ankur; Rashid, Mamoon; Hill-Cawthorne, Grant A.; Ali, Shahjahan; Naeem, Raeece; Thomson, Nicholas R.; Dougan, Gordon; Pain, Arnab

    2014-01-01

    Background:Cholera infection continues to be a threat to global public health. The current cholera pandemic associated with Vibrio cholerae El Tor has now been ongoing for over half a century.Methodology/Principal Findings:Thirty-eight V. cholerae El Tor isolates associated with a cholera outbreak in 2009 from the Chandigarh region of India were characterised by a combination of microbiology, molecular typing and whole-genome sequencing. The genomic analysis indicated that two clones of V. cholera circulated in the region and caused disease during this time. These clones fell into two distinct sub-clades that map independently onto wave 3 of the phylogenetic tree of seventh pandemic V. cholerae El Tor. Sequence analyses of the cholera toxin gene, the Vibrio seventh Pandemic Island II (VSPII) and SXT element correlated with this phylogenetic position of the two clades on the El Tor tree. The clade 2 isolates, characterized by a drug-resistant profile and the expression of a distinct cholera toxin, are closely related to the recent V. cholerae isolated elsewhere, including Haiti, but fell on a distinct branch of the tree, showing they were independent outbreaks. Multi-Locus Sequence Typing (MLST) distinguishes two sequence types among the 38 isolates, that did not correspond to the clades defined by whole-genome sequencing. Multi-Locus Variable-length tandem-nucleotide repeat Analysis (MLVA) identified 16 distinct clusters.Conclusions/Significance:The use of whole-genome sequencing enabled the identification of two clones of V. cholerae that circulated during the 2009 Chandigarh outbreak. These clones harboured a similar structure of ICEVchHai1 but differed mainly in the structure of CTX phage and VSPII. The limited capacity of MLST and MLVA to discriminate between the clones that circulated in the 2009 Chandigarh outbreak highlights the value of whole-genome sequencing as a route to the identification of further genetic markers to subtype V. cholerae isolates.

  10. Cholera toxin subunit B-mediated intracellular trafficking of mesoporous silica nanoparticles toward the endoplasmic reticulum

    Science.gov (United States)

    Walker, William Andrew

    In recent decades, pharmaceutical research has led to the development of numerous treatments for human disease. Nanoscale delivery systems have the potential to maximize therapeutic outcomes by enabling target specific delivery of these therapeutics. The intracellular localization of many of these materials however, is poorly controlled, leading to sequestration in degradative cellular pathways and limiting the efficacy of their payloads. Numerous proteins, particularly bacterial toxins, have evolved mechanisms to subvert the degradative mechanisms of the cell. Here, we have investigated a possible strategy for shunting intracellular delivery of encapsulated cargoes from these pathways by modifying mesoporous silica nanoparticles (MSNs) with the well-characterized bacterial toxin Cholera toxin subunit B (CTxB). Using established optical imaging methods we investigated the internalization, trafficking, and subcellular localization of our modified MSNs in an in vitro animal cell model. We then attempted to demonstrate the practical utility of this approach by using CTxB-modified mesoporous silica nanoparticles to deliver propidium iodide, a membrane-impermeant fluorophore.

  11. Comparison of two recombinant systems for expression of cholera toxin B subunit from Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    M Boustanshenas

    2013-01-01

    Full Text Available Purpose: The aim of this study was to assess the production of recombinant cholera toxin B subunit (rCTB protein in two different expression systems (pAE_ctxB and pQE_ctxB constructs in Escherichia coli BL21 (DE3. Materials and Methods: The ctxB fragment was amplified from Vibrio cholerae O 1 ATCC14035 and cloned in pGETM-T easy vector after which it was transformed to E. coli Top 10F′ and grown on LB-ampicillin agar medium. Sequence analysis confirmed the complete ctxB gene sequence in the construct which was further subcloned to pQE-30 vector. The construct was subsequently transformed to E. coli M15 (pREP4. The recombinant pAE_ctxB and pQE_ctxB were transformed to competent E. coli BL21 (DE3 cells to express CTB protein. Result: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE analysis showed the maximum expression of rCTB in both systems at 5 h after induction and western blot analysis confirmed the presence of recombinant CTB in blotting membranes. Conclusion: Expression of rCTB in pAE_ctxB construct was more efficient (15-fold than pQE_ctxB, and it seems that Lac UV5 in E. coli BL21 (DE3 is more compatible with the former construct. This expression system can be used to produce recombinant CTB in high yield which may enable us to study the oral tolerance or mucosal adjuvant properties of rCTB using animal models.

  12. A Nanocoaxial-Based Electrochemical Sensor for the Detection of Cholera Toxin

    Science.gov (United States)

    Archibald, Michelle; Rizal, Binod; Connolly, Timothy; Burns, Michael J.; Naughton, Michael J.; Chiles, Thomas C.; Biology; Physics Collaboration

    We report a nanocoax-based electrochemical sensor for the detection of bacterial toxins using an electrochemical enzyme-linked immunosorbent assay (ELISA) and differential pulse voltammetry (DPV). The device architecture is composed of vertically-oriented, nanoscale coaxial electrodes, with coax cores and shields serving as integrated working and counter electrodes, respectively. Proof-of-concept was demonstrated for the detection of cholera toxin (CT), with a linear dynamic range of detection was 10 ng/ml - 1 µg/ml, and a limit of detection (LOD) of 2 ng/ml. This level of sensitivity is comparable to the standard optical ELISA used widely in clinical applications. The nanocoax array thus matches the detection profile of the standard ELISA while providing a simple electrochemical readout and a miniaturized platform with multiplexing capabilities, toward point-of-care (POC) implementation. In addition, next generation nanocoax devices with extended cores are currently under development, which would provide a POC platform amenable for biofunctionalization of ELISA receptor proteins directly onto the device. This work was supported by the National Institutes of Health (National Cancer Institute Award No. CA137681 and National Institute of Allergy and Infectious Diseases Award No. AI100216).

  13. Cell Vacuolation Caused by Vibrio cholerae Hemolysin

    Science.gov (United States)

    Figueroa-Arredondo, Paula; Heuser, John E.; Akopyants, Natalia S.; Morisaki, J. Hiroshi; Giono-Cerezo, Silvia; Enríquez-Rincón, Fernando; Berg, Douglas E.

    2001-01-01

    Non-O1 strains of Vibrio cholerae implicated in gastroenteritis and diarrhea generally lack virulence determinants such as cholera toxin that are characteristic of epidemic strains; the factors that contribute to their virulence are not understood. Here we report that at least one-third of diarrhea-associated nonepidemic V. cholerae strains from Mexico cause vacuolation of cultured Vero cells. Detailed analyses indicated that this vacuolation was related to that caused by aerolysin, a pore-forming toxin of Aeromonas; it involved primarily the endoplasmic reticulum at early times (∼1 to 4 h after exposure), and resulted in formation of large, acidic, endosome-like multivesicular vacuoles (probably autophagosomes) only at late times (∼16 h). In contrast to vacuolation caused by Helicobacter pylori VacA protein, that induced by V. cholerae was exacerbated by agents that block vacuolar proton pumping but not by endosome-targeted weak bases. It caused centripetal redistribution of endosomes, reflecting cytoplasmic alkalinization. The gene for V. cholerae vacuolating activity was cloned and was found to correspond to hlyA, the structural gene for hemolysin. HlyA protein is a pore-forming toxin that causes ion leakage and, ultimately, eukaryotic cell lysis. Thus, a distinct form of cell vacuolation precedes cytolysis at low doses of hemolysin. We propose that this vacuolation, in itself, contributes to the virulence of V. cholerae strains, perhaps by perturbing intracellular membrane trafficking or ion exchange in target cells and thereby affecting local intestinal inflammatory or other defense responses. PMID:11179335

  14. Cell vacuolation caused by Vibrio cholerae hemolysin.

    Science.gov (United States)

    Figueroa-Arredondo, P; Heuser, J E; Akopyants, N S; Morisaki, J H; Giono-Cerezo, S; Enríquez-Rincón, F; Berg, D E

    2001-03-01

    Non-O1 strains of Vibrio cholerae implicated in gastroenteritis and diarrhea generally lack virulence determinants such as cholera toxin that are characteristic of epidemic strains; the factors that contribute to their virulence are not understood. Here we report that at least one-third of diarrhea-associated nonepidemic V. cholerae strains from Mexico cause vacuolation of cultured Vero cells. Detailed analyses indicated that this vacuolation was related to that caused by aerolysin, a pore-forming toxin of Aeromonas; it involved primarily the endoplasmic reticulum at early times (approximately 1 to 4 h after exposure), and resulted in formation of large, acidic, endosome-like multivesicular vacuoles (probably autophagosomes) only at late times (approximately 16 h). In contrast to vacuolation caused by Helicobacter pylori VacA protein, that induced by V. cholerae was exacerbated by agents that block vacuolar proton pumping but not by endosome-targeted weak bases. It caused centripetal redistribution of endosomes, reflecting cytoplasmic alkalinization. The gene for V. cholerae vacuolating activity was cloned and was found to correspond to hlyA, the structural gene for hemolysin. HlyA protein is a pore-forming toxin that causes ion leakage and, ultimately, eukaryotic cell lysis. Thus, a distinct form of cell vacuolation precedes cytolysis at low doses of hemolysin. We propose that this vacuolation, in itself, contributes to the virulence of V. cholerae strains, perhaps by perturbing intracellular membrane trafficking or ion exchange in target cells and thereby affecting local intestinal inflammatory or other defense responses.

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

    Directory of Open Access Journals (Sweden)

    Verena Olivier

    2009-10-01

    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.

  16. Oral administration of a recombinant cholera toxin B subunit promotes mucosal healing in the colon.

    Science.gov (United States)

    Baldauf, K J; Royal, J M; Kouokam, J C; Haribabu, B; Jala, V R; Yaddanapudi, K; Hamorsky, K T; Dryden, G W; Matoba, N

    2017-07-01

    Cholera toxin B subunit (CTB) is a component of a licensed oral cholera vaccine. However, CTB has pleiotropic immunomodulatory effects whose impacts on the gut are not fully understood. Here, we found that oral administration in mice of a plant-made recombinant CTB (CTBp) significantly increased several immune cell populations in the colon lamina propria. Global gene expression analysis revealed that CTBp had more pronounced impacts on the colon than the small intestine, with significant activation of TGFβ-mediated pathways in the colon epithelium. The clinical relevance of CTBp-induced impacts on colonic mucosa was examined. In a human colon epithelial model using Caco2 cells, CTBp, but not the non-GM1-binding mutant G33D-CTBp, induced TGFβ-mediated wound healing. In a dextran sodium sulfate (DSS) acute colitis mouse model, oral administration of CTBp protected against colon mucosal damage as manifested by mitigated body weight loss, decreased histopathological scores, and blunted escalation of inflammatory cytokine levels while inducing wound healing-related genes. Furthermore, biweekly oral administration of CTBp significantly reduced disease severity and tumorigenesis in the azoxymethane/DSS model of ulcerative colitis and colon cancer. Altogether, these results demonstrate CTBp's ability to enhance mucosal healing in the colon, highlighting its potential application in ulcerative colitis therapy besides cholera vaccination.

  17. Characterization of a clinical Vibrio cholerae O139 isolate from Mexico.

    Science.gov (United States)

    Parveen, Salina; Farrah, Samuel R; Gonzalez-Bonilla, Celia; Zamudio, Altagracia V; Tamplin, Mark L

    2003-01-01

    Pathogenic strains of Vibrio cholerae O139 possess the cholera toxin A subunit (ctxA) gene as well as the gene for toxin co-regulated pili (tcpA). We report the isolation of a ctxA-negative, tcpA-negative V. cholerae O139 strain (INDREI) from a patient in Mexico diagnosed with gastrointestinal illness. Certain phenotypic characteristics of this strain were identical to those of V. cholerae O1 biotype El Tor. Unlike ctxA-positive V. cholerae O139 strains, this strain was sensitive to a wide panel of antibiotics, including ampicillin, chloramphenicol, ciprofloxacin, gentamicin, furazolidone, nalidixic acid, nitrofurantoin, tetracycline, trimethoprim-sulfamethoxazole, and streptomycin, but was resistant to polymyxin B. Ribotype and pulsed-field gel electrophoresis profiles of INDRE1 differed from those of ctxA-positive V. cholerae O139 and other V. cholerae strains. Phenotypic characteristics of the Mexico strain were similar to those reported for V. cholerae O139 isolates from Argentina and Sri Lanka.

  18. 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.

  19. [Synthesis of protective antigens during submerged cultivation of Vibrio cholerae].

    Science.gov (United States)

    Fedorova, V A; Syrova, N A; Gromova, O V; Tershkina, N E; Devdariani, Z L; Dzhaparidze, M N; Meleshchenko, M V; Dobrova, G V; Beliakova, N I; Ermakov, N M; Eliseev, Iu Iu

    2000-01-01

    The effectiveness of dot immunoanalysis for evaluating the dynamics of the synthesis of O-antigen, cholera toxin, neuraminidase, adhesin CFA1 in the process of the reactor cultivation of V. cholerae used for the production of oral chemical cholera vaccine is shown. The established regularities of the synthesis of the protective antigens of V. cholerae in the process of scaled-up cultivation are discussed.

  20. A novel fluorescent retrograde neural tracer: cholera toxin B conjugated carbon dots

    Science.gov (United States)

    Zhou, Nan; Hao, Zeyu; Zhao, Xiaohuan; Maharjan, Suraj; Zhu, Shoujun; Song, Yubin; Yang, Bai; Lu, Laijin

    2015-09-01

    The retrograde neuroanatomical tracing method is a key technique to study the complex interconnections of the nervous system. Traditional tracers have several drawbacks, including time-consuming immunohistochemical or immunofluorescent staining procedures, rapid fluorescence quenching and low fluorescence intensity. Carbon dots (CDs) have been widely used as a fluorescent bio-probe due to their ultrasmall size, excellent optical properties, chemical stability, biocompatibility and low toxicity. Herein, we develop a novel fluorescent neural tracer: cholera toxin B-carbon dot conjugates (CTB-CDs). It can be taken up and retrogradely transported by neurons in the peripheral nervous system of rats. Our results show that CTB-CDs possess high photoluminescence intensity, good optical stability, a long shelf-life and non-toxicity. Tracing with CTB-CDs is a direct and more economical way of performing retrograde labelling experiments. Therefore, CTB-CDs are reliable fluorescent retrograde tracers.The retrograde neuroanatomical tracing method is a key technique to study the complex interconnections of the nervous system. Traditional tracers have several drawbacks, including time-consuming immunohistochemical or immunofluorescent staining procedures, rapid fluorescence quenching and low fluorescence intensity. Carbon dots (CDs) have been widely used as a fluorescent bio-probe due to their ultrasmall size, excellent optical properties, chemical stability, biocompatibility and low toxicity. Herein, we develop a novel fluorescent neural tracer: cholera toxin B-carbon dot conjugates (CTB-CDs). It can be taken up and retrogradely transported by neurons in the peripheral nervous system of rats. Our results show that CTB-CDs possess high photoluminescence intensity, good optical stability, a long shelf-life and non-toxicity. Tracing with CTB-CDs is a direct and more economical way of performing retrograde labelling experiments. Therefore, CTB-CDs are reliable fluorescent retrograde

  1. Impact of solar irradiation on cholera toxin secretion by different strains of Vibrio cholerae

    CSIR Research Space (South Africa)

    Ssemakalu, CC

    2013-09-01

    Full Text Available , Salomon RN, Garrity K, Reveillaud I, Kopin A, Jackson FR, et al. Vibrio cholerae infection of Drosophila melanogaster mimics the human disease cholera. PLoS Pathog. 2005;1(1):e8. http://dx.doi.org/10.1371/ journal.ppat.0010008 2. World Health...

  2. Quorum-sensing regulators control virulence gene expression in Vibrio cholerae

    OpenAIRE

    Zhu, Jun; Miller, Melissa B.; Vance, Russell E.; Dziejman, Michelle; Bassler, Bonnie L.; Mekalanos, John J.

    2002-01-01

    The production of virulence factors including cholera toxin and the toxin-coregulated pilus in the human pathogen Vibrio cholerae is strongly influenced by environmental conditions. The well-characterized ToxR signal transduction cascade is responsible for sensing and integrating the environmental information and controlling the virulence regulon. We show here that, in addition to the known components of the ToxR signaling circuit, quorum-sensing regulators are involved in regulation of V. ch...

  3. Cholera Toxin Promotes Th17 Cell Differentiation by Modulating Expression of Polarizing Cytokines and the Antigen-Presenting Potential of Dendritic Cells.

    Science.gov (United States)

    Kang, Jung-Ok; Lee, Jee-Boong; Chang, Jun

    2016-01-01

    Cholera toxin (CT), an exotoxin produced by Vibrio cholera, acts as a mucosal adjuvant. In a previous study, we showed that CT skews differentiation of CD4 T cells to IL-17-producing Th17 cells. Here, we found that intranasal administration of CT induced migration of migratory dendritic cell (DC) populations, CD103+ DCs and CD11bhi DCs, to the lung draining mediastinal lymph nodes (medLN). Among those DC subsets, CD11bhi DCs that were relatively immature had a major role in Th17 cell differentiation after administration of CT. CT-treated BMDCs showed reduced expression of MHC class II and CD86, similar to CD11bhi DCs in medLN, and these BMDCs promoted Th17 cell differentiation more potently than other BMDCs expressing higher levels of MHC class II and CD86. By analyzing the expression of activation markers such as CD25 and CD69, proliferation and IL-2 production, we determined that CT-treated BMDCs showed diminished antigen-presenting potential to CD4+ T cells compared with normal BMDCs. We also found that CT-stimulated BMDCs promote activin A expression as well as IL-6 and IL-1β, and activin A had a synergic role with TGF-β1 in CT-mediated Th17 cell differentiation. Taken together, our results suggest that CT-stimulated DCs promote Th17 cell differentiation by not only modulating antigen-presenting potential but also inducing Th polarizing cytokines.

  4. Cholera between 1991 and 1997 in Mexico was associated with infection by classical, El Tor, and El Tor variants of Vibrio cholerae.

    Science.gov (United States)

    Alam, Munirul; Nusrin, Suraia; Islam, Atiqul; Bhuiyan, Nurul A; Rahim, Niaz; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Gil, Ana I; Watanabe, Haruo; Morita, Masatomo; Nair, G Balakrish; Cravioto, Alejandro

    2010-10-01

    Vibrio cholerae O1 biotype El Tor (ET), the cause of the current 7th pandemic, has recently been replaced in Asia and Africa by an altered ET biotype possessing cholera toxin (CTX) of the classical (CL) biotype that originally caused the first six pandemics before becoming extinct in the 1980s. Until recently, the ET prototype was the biotype circulating in Peru; a detailed understanding of the evolutionary trend of V. cholerae causing endemic cholera in Latin America is lacking. The present retrospective microbiological, molecular, and phylogenetic study of V. cholerae isolates recovered in Mexico (n = 91; 1983 to 1997) shows the existence of the pre-1991 CL biotype and the ET and CL biotypes together with the altered ET biotype in both epidemic and endemic cholera between 1991 and 1997. According to sero- and biotyping data, the altered ET, which has shown predominance in Mexico since 1991, emerged locally from ET and CL progenitors that were found coexisting until 1997. In Latin America, ET and CL variants shared a variable number of phenotypic markers, while the altered ET strains had genes encoding the CL CTX (CTX(CL)) prophage, ctxB(CL) and rstR(CL), in addition to resident rstR(ET), as the underlying regional signature. The distinct regional fingerprints for ET in Mexico and Peru and their divergence from ET in Asia and Africa, as confirmed by subclustering patterns in a pulsed-field gel electrophoresis (NotI)-based dendrogram, suggest that the Mexico epidemic in 1991 may have been a local event and not an extension of the epidemics occurring in Asia and South America. Finally, the CL biotype reservoir in Mexico is unprecedented and must have contributed to the changing epidemiology of global cholera in ways that need to be understood.

  5. Cholera in pregnancy: Clinical and immunological aspects.

    Science.gov (United States)

    Khan, Ashraful I; Chowdhury, Fahima; Leung, Daniel T; Larocque, Regina C; Harris, Jason B; Ryan, Edward T; Calderwood, Stephen B; Qadri, Firdausi

    2015-10-01

    The objective of this study was to examine the clinical and immunological features of cholera in pregnancy. Women of reproductive age presenting to the icddr,b Dhaka hospital with cholera, and enrolled as part of a larger cohort study, were tested for pregnancy on admission. We compared initial clinical features and immune responses of pregnant patients with non-pregnant female patients at days 2, 7 and 21 after infection. Among reproductive age women enrolled between January 2001 and May 2006, 9.7% (14/144) were pregnant. The duration of diarrhoea prior to admission tended to be higher in pregnant compared to non-pregnant patients (p=0.08), but other clinical characteristics did not differ. Antibody responses to cholera toxin B subunit (CtxB), toxin-coregulated pilus A (TcpA), Vibrio cholerae lipopolysaccharide (LPS), and serum vibriocidal antibody responses, were comparable between pregnant and non-pregnant patients. There were no deaths among the pregnant cases or non-pregnant controls, and no adverse foetal outcomes, including stillbirths, during 21 days of follow up of pregnant cases. To our knowledge, this is the first report of immune responses in pregnant women with cholera. We found that pregnant woman early in pregnancy has comparable clinical illness and subsequent immune responses compared to non-pregnant women. These findings suggest that the evaluation of safety and immunogenicity of oral cholera vaccines in pregnancy should be an area of future investigations. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  6. Influence of γ-radiation on the immunobiological and immunochemical properties of cholera exotoxin. Com. 2. Immunogenic properties of crudi irradiated toxin filtrate

    International Nuclear Information System (INIS)

    Rubtsov, I.V.; Nedugova, G.I.; Samojlenko, I.I.

    1984-01-01

    The effect of ionizing radiation on immunogenic activity of crude cholera exotoxin (filtrate-toxin), which presents initial raw material to prepare native preventive treatment preparation cholergene-anatoxin has been studied. It is stated, that the use of gamma-radiation for the detoxification and sterilization of cholera exotoxin (crude), preserves its immunogenic properties. The observed increase in immunogenic activity, manifested in the reliable increase of antitoxic antibodies titre to the irradiation preparation over the whole period of observation, as compared with control, in the authors' assumption can be caused by the processes of polymerization and aggregation, taking place in protein molecule during irradiation, which results in the growth of molecule dimensions and in immunogenicity increase

  7. Inhibition of cholera toxin and other AB toxins by polyphenolic compounds

    Science.gov (United States)

    All AB-type protein toxins have intracellular targets despite an initial extracellular location. These toxins use different methods to reach the cytosol and have different effects on the target cell. Broad-spectrum inhibitors against AB toxins are therefore hard to develop because the toxins use dif...

  8. 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

  9. Expression of the Native Cholera Toxin B Subunit Gene and Assembly as Functional Oligomers in Transgenic Tobacco Chloroplasts

    Science.gov (United States)

    Daniell, Henry; Lee, Seung-Bum; Panchal, Tanvi; Wiebe, Peter O.

    2012-01-01

    The B subunits of enterotoxigenic Escherichia coli (LTB) and cholera toxin of Vibrio cholerae (CTB) are candidate vaccine antigens. Integration of an unmodified CTB-coding sequence into chloroplast genomes (up to 10,000 copies per cell), resulted in the accumulation of up to 4.1% of total soluble tobacco leaf protein as functional oligomers (410-fold higher expression levels than that of the unmodified LTB gene expressed via the nuclear genome). However, expresssion levels reported are an underestimation of actual accumulation of CTB in transgenic chloroplasts, due to aggregation of the oligomeric forms in unboiled samples similar to the aggregation observed for purified bacterial antigen. PCR and Southern blot analyses confirmed stable integration of the CTB gene into the chloroplast genome. Western blot analysis showed that the chloroplast-synthesized CTB assembled into oligomers and were antigenically identical with purified native CTB. Also, binding assays confirmed that chloroplast- synthesized CTB binds to the intestinal membrane GM1-ganglioside receptor, indicating correct folding and disulfide bond formation of CTB pentamers within transgenic chloroplasts. In contrast to stunted nuclear transgenic plants, chloroplast transgenic plants were morphologically indistinguishable from untransformed plants, when CTB was constitutively expressed in chloroplasts. Introduced genes were inherited stably in subsequent generations, as confirmed by PCR and Southern blot analyses. Increased production of an efficient transmucosal carrier molecule and delivery system, like CTB, in transgenic chloroplasts makes plant-based oral vaccines and fusion proteins with CTB needing oral administration commercially feasible. Successful expression of foreign genes in transgenic chromoplasts and availability of marker-free chloroplast transformation techniques augurs well for development of vaccines in edible parts of transgenic plants. Furthermore, since the quaternary structure of

  10. 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.

  11. Genetic markers for western corn rootworm resistance to Bt toxin.

    Science.gov (United States)

    Flagel, Lex E; Swarup, Shilpa; Chen, Mao; Bauer, Christopher; Wanjugi, Humphrey; Carroll, Matthew; Hill, Patrick; Tuscan, Meghan; Bansal, Raman; Flannagan, Ronald; Clark, Thomas L; Michel, Andrew P; Head, Graham P; Goldman, Barry S

    2015-01-07

    Western corn rootworm (WCR) is a major maize (Zea mays L.) pest leading to annual economic losses of more than 1 billion dollars in the United States. Transgenic maize expressing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are widely used for the management of WCR. However, cultivation of Bt-expressing maize places intense selection pressure on pest populations to evolve resistance. Instances of resistance to Bt toxins have been reported in WCR. Developing genetic markers for resistance will help in characterizing the extent of existing issues, predicting where future field failures may occur, improving insect resistance management strategies, and in designing and sustainably implementing forthcoming WCR control products. Here, we discover and validate genetic markers in WCR that are associated with resistance to the Cry3Bb1 Bt toxin. A field-derived WCR population known to be resistant to the Cry3Bb1 Bt toxin was used to generate a genetic map and to identify a genomic region associated with Cry3Bb1 resistance. Our results indicate that resistance is inherited in a nearly recessive manner and associated with a single autosomal linkage group. Markers tightly linked with resistance were validated using WCR populations collected from Cry3Bb1 maize fields showing significant WCR damage from across the US Corn Belt. Two markers were found to be correlated with both diet (R2 = 0.14) and plant (R2 = 0.23) bioassays for resistance. These results will assist in assessing resistance risk for different WCR populations, and can be used to improve insect resistance management strategies. Copyright © 2015 Flagel et al.

  12. The protective activity of tea catechins against experimental infection by Vibrio cholerae O1.

    Science.gov (United States)

    Toda, M; Okubo, S; Ikigai, H; Suzuki, T; Suzuki, Y; Hara, Y; Shimamura, T

    1992-01-01

    Tea catechins inhibited the fluid accumulation induced by cholera toxin in sealed adult mice. The catechins also reduced fluid accumulation by Vibrio cholerae O1 in ligated intestinal loops of rabbits. These findings suggest that tea catechins may possess protective activity against V. cholerae O1.

  13. Molecular Epidemiology of Cholera Outbreaks during the Rainy Season in Mandalay, Myanmar.

    Science.gov (United States)

    Roobthaisong, Amonrattana; Okada, Kazuhisa; Htun, Nilar; Aung, Wah Wah; Wongboot, Warawan; Kamjumphol, Watcharaporn; Han, Aye Aye; Yi, Yi; Hamada, Shigeyuki

    2017-11-01

    Cholera, caused by Vibrio cholerae , remains a global threat to public health. In Myanmar, the availability of published information on the occurrence of the disease is scarce. We report here that cholera incidence in Mandalay generally exhibited a single annual peak, with an annual average of 312 patients with severe dehydration over the past 5 years (since 2011) and was closely associated with the rainy season. We analyzed cholera outbreaks, characterized 67 isolates of V. cholerae serogroup O1 in 2015 from patients from Mandalay, and compared them with 22 V. cholerae O1 isolates (12 from Mandalay and 10 from Yangon) in 2014. The isolates carried the classical cholera toxin B subunit ( ctxB ), the toxin-coregulated pilus A ( tcpA ) of Haitian type, and repeat sequence transcriptional regulator ( rstR ) of El Tor type. Two molecular typing methods, pulsed-field gel electrophoresis and multiple-locus variable-number tandem repeat analysis (MLVA), differentiated the 89 isolates into seven pulsotypes and 15 MLVA profiles. Pulsotype Y15 and one MLVA profile (11, 7, 7, 16, 7) were predominantly found in the isolates from cholera outbreaks in Mandalay, 2015. Pulsotypes Y11, Y12, and Y15 with some MLVA profiles were detected in the isolates from two remote areas, Mandalay and Yangon, with temporal changes. These data suggested that cholera spread from the seaside to the inland area in Myanmar.

  14. Vibrio cholerae infection, novel drug targets and phage therapy.

    Science.gov (United States)

    Fazil, Mobashar Hussain Urf Turabe; Singh, Durg V

    2011-10-01

    Vibrio cholerae is the causative agent of the diarrheal disease cholera. Although antibiotic therapy shortens the duration of diarrhea, excessive use has contributed to the emergence of antibiotic resistance in V. cholerae. Mobile genetic elements have been shown to be largely responsible for the shift of drug resistance genes in bacteria, including some V. cholerae strains. Quorum sensing communication systems are used for interaction among bacteria and for sensing environmental signals. Sequence analysis of the ctxB gene of toxigenic V. cholerae strains demonstrated its presence in multiple cholera toxin genotypes. Moreover, bacteriophage that lyse the bacterium have been reported to modulate epidemics by decreasing the required infectious dose of the bacterium. In this article, we will briefly discuss the disease, its clinical manifestation, antimicrobial resistance and the novel approaches to locate drug targets to treat cholera.

  15. Molecular Analyses of Vibrio cholerae O1 Clinical Strains, Including New Nontoxigenic Variants Isolated in Mexico during the Cholera Epidemic Years between 1991 and 2000

    OpenAIRE

    Lizárraga-Partida, Leonardo; Quilici, Marie-Laure

    2009-01-01

    International audience; We studied the evolution of Vibrio cholerae O1 during the 1991 to 2000 cholera epidemic in Mexico by biochemical, serological, and molecular characterization of strains collected during this period. Strains were divided into toxigenic and nontoxigenic groups according to the presence or absence of genes encoding cholera toxin. As previously reported, we characterized two populations among toxigenic strains, which were present from the first year of the epidemic. BglI r...

  16. Bovine lactoferrin decreases cholera-toxin-induced intestinal fluid accumulation in mice by ganglioside interaction.

    Directory of Open Access Journals (Sweden)

    Fulton P Rivera

    Full Text Available Secretory diarrhea caused by cholera toxin (CT is initiated by binding of CT's B subunit (CTB to GM1-ganglioside on the surface of intestinal cells. Lactoferrin, a breast milk glycoprotein, has shown protective effect against several enteropathogens. The aims of this study were to determine the effect of bovine-lactoferrin (bLF on CT-induced intestinal fluid accumulation in mice, and the interaction between bLF and CT/CTB with the GM1-ganglioside receptor. Fluid accumulation induced by CT was evaluated in the mouse ileal loop model using 56 BALB/c mice, with and without bLF added before, after or at the same time of CT administration. The effect of bLF in the interaction of CT and CTB with GM1-ganglioside was evaluated by a GM1-enzyme-linked immunosorbent assay. bLF decreased CT-induced fluid accumulation in the ileal loop of mice. The greatest effect was when bLF was added before CT (median, 0.066 vs. 0.166 g/cm, with and without bLF respectively, p<0.01. We conclude that bLF decreases binding of CT and CTB to GM1-ganglioside, suggesting that bLF suppresses CT-induced fluid accumulation by blocking the binding of CTB to GM1-ganglioside. bLF may be effective as adjunctive therapy for treatment of cholera diarrhea.

  17. Association of Heavy Rainfall on Genotypic Diversity in V. cholerae Isolates from an Outbreak in India

    Directory of Open Access Journals (Sweden)

    A. K. Goel

    2011-01-01

    Full Text Available The outbreak of waterborne disease cholera has been associated with rainfall and flooding events by contamination of potable water with environmental Vibrio cholerae. The continuation of the epidemic in a region, however, is often due to secondary transmission of the initial outbreak strain through human waste. This paper reports, on the contrary, a rapid shift of genotype from one V. cholerae strain to another one in an epidemic region. V. cholerae isolated from patients during 2005 cholera epidemic in Chennai, India were characterized using PCR identification of toxin genes, antibiogram, and genomic fingerprinting analysis. The results showed that in spite of the similarity of toxin genes and antibiogram, the Vibrio isolates grouped into two different clusters based on the ERIC-PCR fingerprinting. Each cluster corresponded to a distinct peak of cholera outbreak, which occurred after separate heavy rainfall. The results suggest that the rainfall event can bring various genotypes of V. cholerae strains causing multiple outbreaks.

  18. Rapid and scalable plant-based production of a cholera toxin B subunit variant to aid in mass vaccination against cholera outbreaks.

    Directory of Open Access Journals (Sweden)

    Krystal Teasley Hamorsky

    Full Text Available INTRODUCTION: Cholera toxin B subunit (CTB is a component of an internationally licensed oral cholera vaccine. The protein induces neutralizing antibodies against the holotoxin, the virulence factor responsible for severe diarrhea. A field clinical trial has suggested that the addition of CTB to killed whole-cell bacteria provides superior short-term protection to whole-cell-only vaccines; however, challenges in CTB biomanufacturing (i.e., cost and scale hamper its implementation to mass vaccination in developing countries. To provide a potential solution to this issue, we developed a rapid, robust, and scalable CTB production system in plants. METHODOLOGY/PRINCIPAL FINDINGS: In a preliminary study of expressing original CTB in transgenic Nicotiana benthamiana, the protein was N-glycosylated with plant-specific glycans. Thus, an aglycosylated CTB variant (pCTB was created and overexpressed via a plant virus vector. Upon additional transgene engineering for retention in the endoplasmic reticulum and optimization of a secretory signal, the yield of pCTB was dramatically improved, reaching >1 g per kg of fresh leaf material. The protein was efficiently purified by simple two-step chromatography. The GM1-ganglioside binding capacity and conformational stability of pCTB were virtually identical to the bacteria-derived original B subunit, as demonstrated in competitive enzyme-linked immunosorbent assay, surface plasmon resonance, and fluorescence-based thermal shift assay. Mammalian cell surface-binding was corroborated by immunofluorescence and flow cytometry. pCTB exhibited strong oral immunogenicity in mice, inducing significant levels of CTB-specific intestinal antibodies that persisted over 6 months. Moreover, these antibodies effectively neutralized the cholera holotoxin in vitro. CONCLUSIONS/SIGNIFICANCE: Taken together, these results demonstrated that pCTB has robust producibility in Nicotiana plants and retains most, if not all, of major

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

    Science.gov (United States)

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

    2016-01-01

    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

  20. Alga-Produced Cholera Toxin-Pfs25 Fusion Proteins as Oral Vaccines

    Science.gov (United States)

    Gregory, James A.; Topol, Aaron B.; Doerner, David Z.

    2013-01-01

    Infectious diseases disproportionately affect indigent regions and are the greatest cause of childhood mortality in developing countries. Practical, low-cost vaccines for use in these countries are paramount to reducing disease burdens and concomitant poverty. Algae are a promising low-cost system for producing vaccines that can be orally delivered, thereby avoiding expensive purification and injectable delivery. We engineered the chloroplast of the eukaryotic alga Chlamydomonas reinhardtii to produce a chimeric protein consisting of the 25-kDa Plasmodium falciparum surface protein (Pfs25) fused to the β subunit of the cholera toxin (CtxB) to investigate an alga-based whole-cell oral vaccine. Pfs25 is a promising malaria transmission-blocking vaccine candidate that has been difficult to produce in traditional recombinant systems due to its structurally complex tandem repeats of epidermal growth factor-like domains. The noncatalytic CtxB domain of the cholera holotoxin assembles into a pentameric structure and acts as a mucosal adjuvant by binding GM1 ganglioside receptors on gut epithelial cells. We demonstrate that CtxB-Pfs25 accumulates as a soluble, properly folded and functional protein within algal chloroplasts, and it is stable in freeze-dried alga cells at ambient temperatures. In mice, oral vaccination using freeze-dried algae that produce CtxB-Pfs25 elicited CtxB-specific serum IgG antibodies and both CtxB- and Pfs25-specific secretory IgA antibodies. These data suggest that algae are a promising system for production and oral delivery of vaccine antigens, but as an orally delivered adjuvant, CtxB is best suited for eliciting secretory IgA antibodies for vaccine antigens against pathogens that invade mucosal surfaces using this strategy. PMID:23603678

  1. Small-molecule inhibitors of toxT expression in Vibrio cholerae.

    Science.gov (United States)

    Anthouard, Rebecca; DiRita, Victor J

    2013-08-06

    Vibrio cholerae, a Gram-negative bacterium, infects humans and causes cholera, a severe disease characterized by vomiting and diarrhea. These symptoms are primarily caused by cholera toxin (CT), whose production by V. cholerae is tightly regulated by the virulence cascade. In this study, we designed and carried out a high-throughput chemical genetic screen to identify inhibitors of the virulence cascade. We identified three compounds, which we named toxtazin A and toxtazin B and B', representing two novel classes of toxT transcription inhibitors. All three compounds reduce production of both CT and the toxin-coregulated pilus (TCP), an important colonization factor. We present evidence that toxtazin A works at the level of the toxT promoter and that toxtazins B and B' work at the level of the tcpP promoter. Treatment with toxtazin B results in a 100-fold reduction in colonization in an infant mouse model of infection, though toxtazin A did not reduce colonization at the concentrations tested. These results add to the growing body of literature indicating that small-molecule inhibitors of virulence genes could be developed to treat infections, as alternatives to antibiotics become increasingly needed. V. cholerae caused more than 580,000 infections worldwide in 2011 alone (WHO, Wkly. Epidemiol. Rec. 87:289-304, 2012). Cholera is treated with an oral rehydration therapy consisting of water, glucose, and electrolytes. However, as V. cholerae is transmitted via contaminated water, treatment can be difficult for communities whose water source is contaminated. In this study, we address the need for new therapeutic approaches by targeting the production of the main virulence factor, cholera toxin (CT). The high-throughput screen presented here led to the identification of two novel classes of inhibitors of the virulence cascade in V. cholerae, toxtazin A and toxtazins B and B'. We demonstrate that (i) small-molecule inhibitors of virulence gene production can be

  2. Effect of cholera toxin on cAMP levels and Na+ influx in isolated intestinal epithelial cells

    International Nuclear Information System (INIS)

    Hyun, C.S.; Kimmich, G.A.

    1982-01-01

    Freshly isolated chicken intestinal cells contain approximately 20 pmol adenosine 3',5'-cyclic monophosphate (cAMP)/mg cellular protein. Incubation with 3 μg/ml cholera toxin (CT) at 37 0 C induces an elevation of cellular cAMP beginning 10-15 min after initial exposure. The response is linear with time for 40-50 min and causes a six- to eightfold increase over control levels at steady state. Dibutyryl cAMP and agents that increase cAMP production inhibit Na + influx into the isolated enterocytes. Chlorpromazine completely abolishes the toxin-induced elevation of cAMP in the isolated cells and also reverses the effect on Na + entry. The data provide evidence for a cAMP-mediated control of intestinal cell Na + uptake, which may represent the mechanistic basis for the antiabsorptive effect of CT on Na + during induction of intestinal secretory activity. Studies on the time-dependent effects of chlorpromazine on both intracellular cAMP concentration and Na + influx suggest that the reactivation of the Na + transport system after cAMP-induced inhibition is slow relative to the disappearance of cAMP

  3. Structure of Vibrio cholerae ToxT reveals a mechanism for fatty acid regulation of virulence genes

    Energy Technology Data Exchange (ETDEWEB)

    Lowden, Michael J.; Skorupski, Karen; Pellegrini, Maria; Chiorazzo, Michael G.; Taylor, Ronald K.; Kull, F. Jon (Dartmouth)

    2010-03-04

    Cholera is an acute intestinal infection caused by the bacterium Vibrio cholerae. In order for V. cholerae to cause disease, it must produce two virulence factors, the toxin-coregulated pilus (TCP) and cholera toxin (CT), whose expression is controlled by a transcriptional cascade culminating with the expression of the AraC-family regulator, ToxT. We have solved the 1.9 {angstrom} resolution crystal structure of ToxT, which reveals folds in the N- and C-terminal domains that share a number of features in common with AraC, MarA, and Rob as well as the unexpected presence of a buried 16-carbon fatty acid, cis-palmitoleate. The finding that cis-palmitoleic acid reduces TCP and CT expression in V. cholerae and prevents ToxT from binding to DNA in vitro provides a direct link between the host environment of V. cholerae and regulation of virulence gene expression.

  4. N-Glycosylation of cholera toxin B subunit: serendipity for novel plant-made vaccines?

    Directory of Open Access Journals (Sweden)

    Nobuyuki eMatoba

    2015-12-01

    Full Text Available The non-toxic B subunit of cholera toxin (CTB has attracted considerable interests from vaccinologists due to strong mucosal immunomodulatory effects and potential utility as a vaccine scaffold for heterologous antigens. Along with other conventional protein expression systems, various plant species have been used as recombinant production hosts for CTB and its fusion proteins. However, it has recently become clear that the protein is N-glycosylated within the endoplasmic reticulum of plant cells – a eukaryotic post-translational modification that is not present in native CTB. While functionally active aglycosylated variants have been successfully engineered to circumvent potential safety and regulatory issues related to glycosylation, this modification may actually provide advantageous characteristics to the protein as a vaccine platform. Based on data from our recent studies, I discuss the unique features of N-glycosylated CTB produced in plants for the development of novel vaccines.

  5. 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.

  6. Nanodiscs for immobilization of lipid bilayers and membrane receptors: kinetic analysis of cholera toxin binding to a glycolipid receptor

    DEFF Research Database (Denmark)

    Borch, Jonas; Torta, Federico; Sligar, Stephen G

    2008-01-01

    nanodiscs and their incorporated membrane receptors can be attached to surface plasmon resonance sensorchips and used to measure the kinetics of the interaction between soluble molecules and membrane receptors inserted in the bilayer of nanodiscs. Cholera toxin and its glycolipid receptor G(M1) constitute...... a system that can be considered a paradigm for interactions of soluble proteins with membrane receptors. In this work, we have investigated different technologies for capturing nanodiscs containing the glycolipid receptor G(M1) in lipid bilayers, enabling measurements of binding of its soluble interaction...

  7. 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

    . 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...... 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...

  8. The molecular epidemiology of cholera in Latin America.

    Science.gov (United States)

    Wachsmuth, I K; Evins, G M; Fields, P I; Olsvik, O; Popovic, T; Bopp, C A; Wells, J G; Carrillo, C; Blake, P A

    1993-03-01

    To explain the sudden appearance and rapid spread of cholera in Latin America in January 1991, molecular techniques were used to define Vibrio cholerae O1 isolates from around the world. Restriction fragment length polymorphisms of rRNA and ctxA genes, DNA sequence of cholera toxin B subunit gene ctxB, and multilocus enzyme electrophoresis data were used to characterize 197 isolates. Worldwide, there are at least four distinct toxigenic El Tor V. cholerae O1 clones: the seventh pandemic (Eastern Hemisphere), US Gulf Coast, Australian, and Latin American. Nontoxigenic V. cholerae O1 previously isolated in Brazil, Mexico, and Peru are unlike current toxigenic isolates. The Latin American clone probably represents an extension of the seventh pandemic into the Western Hemisphere, while the US Gulf Coast clone most likely evolved separately. These data will be useful in monitoring the spread of cholera, determining the origin of outbreaks in both hemispheres, and implicating specific vehicles of transmission.

  9. Increased jejunal prostaglandin E2 concentrations in patients with acute cholera

    NARCIS (Netherlands)

    Speelman, P.; Rabbani, G. H.; Bukhave, K.; Rask-Madsen, J.

    1985-01-01

    Supraphysiologic doses of prostaglandins (PGs) mimic the effect of cholera toxin and cAMP in the small intestine, but not all observations are explicable in terms of the theory that links PGs to cAMP. Because no data exist on endogenous PGs in human cholera we measured PGE2 concentrations in jejunal

  10. Influence of bacterial toxins on the GTPase activity of transducin from bovine retinal rod outer segments

    International Nuclear Information System (INIS)

    Rybin, V.O.; Gureeva, A.A.

    1986-01-01

    The action of cholera toxin, capable of ADP-ribosylation of the activator N/sub s/ protein, and pertussis toxin, capable of ADP-ribosylation of the inhibitor N/sub i/ protein of the adenylate cyclase complex, on transducin, the GTP-binding protein of the rod outer segments of the retina, was investigated. It was shown that under the action of pertussis and cholera toxins, the GTPase activity of transducin is inhibited. Pertussin toxin inhibits the GTPase of native retinal rod outer segments by 30-40%, while GTPase of homogeneous transducin produces a 70-80% inhibition. The action of toxins on transducin depends on the presence and nature of the guanylic nucleotide with which incubation is performed. On the basis of the data obtained it is suggested that pertussis toxin interacts with pretransducin and with the transducin-GDP complex, while cholera toxin ADP-ribosylates the transducin-GTP complex and does not act on transducin lacking GTP

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

    Directory of Open Access Journals (Sweden)

    Goddeeris Bruno M

    2009-12-01

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

  12. 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.

  13. Riverbed Sediments as Reservoirs of Multiple Vibrio cholerae Virulence-Associated Genes: A Potential Trigger for Cholera Outbreaks in Developing Countries.

    Science.gov (United States)

    Abia, Akebe Luther King; Ubomba-Jaswa, Eunice; Momba, Maggy Ndombo Benteke

    2017-01-01

    Africa remains the most cholera stricken continent in the world as many people lacking access to safe drinking water rely mostly on polluted rivers as their main water sources. However, studies in these countries investigating the presence of Vibrio cholerae in aquatic environments have paid little attention to bed sediments. Also, information on the presence of virulence-associated genes (VAGs) in environmental ctx -negative V. cholerae strains in this region is lacking. Thus, we investigated the presence of V. cholerae VAGs in water and riverbed sediment of the Apies River, South Africa. Altogether, 120 samples (60 water and 60 sediment samples) collected from ten sites on the river (January and February 2014) were analysed using PCR. Of the 120 samples, 37 sediment and 31 water samples were positive for at least one of the genes investigated. The haemolysin gene (hlyA) was the most isolated gene. The cholera toxin (ctxAB) and non-O1 heat-stable (stn/sto) genes were not detected. Genes were frequently detected at sites influenced by human activities. Thus, identification of V. cholerae VAGs in sediments suggests the possible presence of V. cholerae and identifies sediments of the Apies River as a reservoir for potentially pathogenic V. cholerae with possible public health implications.

  14. Riverbed Sediments as Reservoirs of Multiple Vibrio cholerae Virulence-Associated Genes: A Potential Trigger for Cholera Outbreaks in Developing Countries

    Directory of Open Access Journals (Sweden)

    Akebe Luther King Abia

    2017-01-01

    Full Text Available Africa remains the most cholera stricken continent in the world as many people lacking access to safe drinking water rely mostly on polluted rivers as their main water sources. However, studies in these countries investigating the presence of Vibrio cholerae in aquatic environments have paid little attention to bed sediments. Also, information on the presence of virulence-associated genes (VAGs in environmental ctx-negative V. cholerae strains in this region is lacking. Thus, we investigated the presence of V. cholerae VAGs in water and riverbed sediment of the Apies River, South Africa. Altogether, 120 samples (60 water and 60 sediment samples collected from ten sites on the river (January and February 2014 were analysed using PCR. Of the 120 samples, 37 sediment and 31 water samples were positive for at least one of the genes investigated. The haemolysin gene (hlyA was the most isolated gene. The cholera toxin (ctxAB and non-O1 heat-stable (stn/sto genes were not detected. Genes were frequently detected at sites influenced by human activities. Thus, identification of V. cholerae VAGs in sediments suggests the possible presence of V. cholerae and identifies sediments of the Apies River as a reservoir for potentially pathogenic V. cholerae with possible public health implications.

  15. ToxR regulates the production of lipoproteins and the expression of serum resistance in Vibrio cholerae

    International Nuclear Information System (INIS)

    Parsot, C.; Taxman, E.; Mekalanos, J.J.

    1991-01-01

    The genes encoding three lipoproteins of Vibrio cholerae were identified by a combination of DNA sequence analysis and [ 3 H]palmitate labeling of hybrid proteins encoded by TnphoA gene fusions. The expression of these three lipoproteins, TagA, AcfD, and TcpC, was controlled by ToxR, the cholera toxin transcriptional activator. The involvement of other bacterial lipoproteins in conferring resistance ot the bactericidal effects of complement prompted us to examine this possibility in V. cholerae. Remarkably, mutations in toxR and tcp genes (including tcpC), involved in the biogenesis of the toxin coregulated pili, rendered V. cholerae about 10 4 - 10 6 times more sensitive to the vibriocidal activity of antibody and complement. Since V. cholerae is a noninvasive organism and toxR and tcp mutants are highly defective in intestinal colonization in animals and humans, these results raise the possibility that resistance to a gut-associated, complement-like bactericidal activity may be a major virulence determinant of V. cholerae and other enterobacterial species

  16. Effect of small doses of ionizing radiation on motility, rosette formation, and antioxidant state of leukocytes under modification of G-proteins by cholera and pertussis toxins

    International Nuclear Information System (INIS)

    Zhirnov, V.V.; Luik, A.I.; Metelitsa, L.A.; Mogilevich, S.E.; Charochkina, L.L.

    2000-01-01

    The responses of motility and rosette formation of leukocytes to small radioactive doses (from 6 centre dot 10 -10 to 6 centre dot 10 -4 Gy) are studied. The influence of these doses on cell functions and oxidative homeostasis are investigated under the modification of transducing components of membrane signal pathways (adenylate cyclase and polyphosphoinositide cascades) with pertussis and cholera toxins

  17. Acalculous cholecystitis and septicemia caused by non-O1 Vibrio cholerae: first reported case and review of biliary infections with Vibrio cholerae.

    Science.gov (United States)

    West, B C; Silberman, R; Otterson, W N

    1998-03-01

    The first case of septicemic acute acalculous cholecystitis caused by non-O1 Vibrio cholerae is described in a healthy traveler, and biliary tract infections from V. cholerae are reviewed. Immediately after a vacation in Cancun, Mexico, a 55-year-old man developed acute cholecystitis. Blood and bile cultures grew non-O1 V. cholerae. At surgery, the gallbladder was acalculous, inflamed, distended, and nearly ruptured. Pathogenetic factors may have included diarrhea prophylaxis with bismuth subsalicylate, distension of the gallbladder from illness-induced fasting, and bacterial toxins in the gallbladder. The patient received i.v. cephapirin, followed by oral cephradine for a total of 10 days, and he made a quick and complete recovery. V. cholerae should be considered in the differential diagnosis of persons from endemic areas who present with cholecystitis or acute jaundice.

  18. Effect of Cholera Toxin Administered Supraspinally or Spinally on the Blood Glucose Level in Pain and D-Glucose Fed Animal Models

    OpenAIRE

    Sim, Yun-Beom; Park, Soo-Hyun; Kang, Yu-Jung; Kim, Sung-Su; Kim, Chea-Ha; Kim, Su-Jin; Jung, Jun-Sub; Ryu, Ohk-Hyun; Choi, Moon-Gi; Choi, Seong-Soo; Suh, Hong-Won

    2013-01-01

    In the present study, the effect of intrathecal (i.t.) or intracerebroventricular (i.c.v.) administration with cholera toxin (CTX) on the blood glucose level was examined in ICR mice. The i.t. treatment with CTX alone for 24 h dose-dependently increased the blood glucose level. However, i.c.v. treatment with CTX for 24 h did not affect the blood glucose level. When mice were orally fed with D-glucose (2 g/kg), the blood glucose level reached to a maximum level at 30 min and almost returned to...

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

    Science.gov (United States)

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

    2014-07-08

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

  20. Molecular analyses of Vibrio cholerae O1 clinical strains, including new nontoxigenic variants isolated in Mexico during the Cholera epidemic years between 1991 and 2000.

    Science.gov (United States)

    Lizárraga-Partida, Marcial Leonardo; Quilici, Marie-Laure

    2009-05-01

    We studied the evolution of Vibrio cholerae O1 during the 1991 to 2000 cholera epidemic in Mexico by biochemical, serological, and molecular characterization of strains collected during this period. Strains were divided into toxigenic and nontoxigenic groups according to the presence or absence of genes encoding cholera toxin. As previously reported, we characterized two populations among toxigenic strains, which were present from the first year of the epidemic. BglI rRNA analysis revealed that these strains had ribotype profiles, denoted M5 and M6 in our study, that were identical to those previously designated Koblavi B5 or Popovic 5 and Popovic 6a or Tamayo B21a, respectively. Ribotype M5 was isolated between 1991 and 1993. This ribotype had a low level of genetic variation as detected by pulsed-field gel electrophoresis (PFGE). Ribotype M6 persisted from 1991 to 2000. However, PFGE profiles suggested that two epidemiologically unrelated strains coexisted within this single ribotype from 1995 until the end of the epidemic. We identified three new BglI ribotypes, Mx1, Mx2, and Mx3, from nontoxigenic V. cholerae O1 strains isolated between 1998 and 2000; one of them grouped strains positive for the toxin-coregulated pilus island. They differed from nontoxigenic clones isolated in Latin America and on the U.S. Gulf Coast and are probably autochthonous Mexican V. cholerae O1 variants. Most of these new variants were isolated from states surrounding the Gulf of Mexico, where the highest incidence of cholera in the country was recorded. Thus, the Mexican Gulf Coast, like the U.S. Gulf Coast, may act as an environmental reservoir of V. cholerae O1.

  1. Application and Development of Biological AFM for the Study of Bacterial Toxins

    National Research Council Canada - National Science Library

    Yang, Jie

    1999-01-01

    ... with other conventional methods. These studies have also established a solid foundation for our structural elucidation of molecular level conformation of membranous bacterial toxins, such as cholera toxin and alpha-hemolysin...

  2. Rotavirus 2/6 Viruslike Particles Administered Intranasally with Cholera Toxin, Escherichia coli Heat-Labile Toxin (LT), and LT-R192G Induce Protection from Rotavirus Challenge

    Science.gov (United States)

    O’Neal, Christine M.; Clements, John D.; Estes, Mary K.; Conner, Margaret E.

    1998-01-01

    We have shown that rotavirus 2/6 viruslike particles composed of proteins VP2 and VP6 (2/6-VLPs) administered to mice intranasally with cholera toxin (CT) induced protection from rotavirus challenge, as measured by virus shedding. Since it is unclear if CT will be approved for human use, we evaluated the adjuvanticity of Escherichia coli heat-labile toxin (LT) and LT-R192G. Mice were inoculated intranasally with 10 μg of 2/6-VLPs combined with CT, LT, or LT-R192G. All three adjuvants induced equivalent geometric mean titers of rotavirus-specific serum antibody and intestinal immunoglobulin G (IgG). Mice inoculated with 2/6-VLPs with LT produced significantly higher titers of intestinal IgA than mice given CT as the adjuvant. All mice inoculated with 2/6-VLPs mixed with LT and LT-R192G were totally protected (100%) from rotavirus challenge, while mice inoculated with 2/6-VLPs mixed with CT showed a mean 91% protection from challenge. The availability of a safe, effective mucosal adjuvant such as LT-R192G will increase the practicality of administering recombinant vaccines mucosally. PMID:9525668

  3. A Mutational Analysis of Residues in Cholera Toxin A1 Necessary for Interaction with Its Substrate, the Stimulatory G Protein Gsα

    Directory of Open Access Journals (Sweden)

    Michael G. Jobling

    2015-03-01

    Full Text Available Pathogenesis of cholera diarrhea requires cholera toxin (CT-mediated adenosine diphosphate (ADP-ribosylation of stimulatory G protein (Gsα in enterocytes. CT is an AB5 toxin with an inactive CTA1 domain linked via CTA2 to a pentameric receptor-binding B subunit. Allosterically activated CTA1 fragment in complex with NAD+ and GTP-bound ADP-ribosylation factor 6 (ARF6-GTP differs conformationally from the CTA1 domain in holotoxin. A surface-exposed knob and a short α-helix (formed, respectively, by rearranging “active-site” and “activation” loops in inactive CTA1 and an ADP ribosylating turn-turn (ARTT motif, all located near the CTA1 catalytic site, were evaluated for possible roles in recognizing Gsα. CT variants with one, two or three alanine substitutions at surface-exposed residues within these CTA1 motifs were tested for assembly into holotoxin and ADP-ribosylating activity against Gsα and diethylamino-(benzylidineamino-guanidine (DEABAG, a small substrate predicted to fit into the CTA1 active site. Variants with single alanine substitutions at H55, R67, L71, S78, or D109 had nearly wild-type activity with DEABAG but significantly decreased activity with Gsα, suggesting that the corresponding residues in native CTA1 participate in recognizing Gsα. As several variants with multiple substitutions at these positions retained partial activity against Gsα, other residues in CTA1 likely also participate in recognizing Gsα.

  4. Influence of gamma radiation on the immunobiological and immunochemical properties of cholera exotoxin. Communication 1. Changes in the biological activity of crude cholera exotoxin under the action of ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Nedugova, G I; Rubtsov, I V; Samojlenko, I I [Ministerstvo Zdravookhraneniya SSSR, Tsentral' nyj Inst. Ehpidemiologii

    1984-02-01

    Native cholera exotoxin (abacterial centrifugalized deposit) has been irradiated using gamma-installations with a /sup 60/Co source. A high inactivating effect of gamma-radiation on native cholera exotoxin is established: with the increase of radiation dose cholerogenity decreased for certain (at the dose 50 kGy) a complete inactivation of all studied series of liquid filtrate-toxin took place), activity of permeability factor and toxicity for mice decreased. A higher radiostability of dry toxin preparations as compared with the liquid ones is detected. Sterilization effect of radiation is achieved at the dose 20 kGy for liquid preparations and at the dose of 30 kGy for dry ones. When preserving the irradiated preparations of raw toxin in different temperature regimes for 6 months to 1.5 year (observation time) toxic properties are not restored, immunogenous properties do not change.

  5. Molecular Analyses of Vibrio cholerae O1 Clinical Strains, Including New Nontoxigenic Variants Isolated in Mexico during the Cholera Epidemic Years between 1991 and 2000▿

    Science.gov (United States)

    Lizárraga-Partida, Marcial Leonardo; Quilici, Marie-Laure

    2009-01-01

    We studied the evolution of Vibrio cholerae O1 during the 1991 to 2000 cholera epidemic in Mexico by biochemical, serological, and molecular characterization of strains collected during this period. Strains were divided into toxigenic and nontoxigenic groups according to the presence or absence of genes encoding cholera toxin. As previously reported, we characterized two populations among toxigenic strains, which were present from the first year of the epidemic. BglI rRNA analysis revealed that these strains had ribotype profiles, denoted M5 and M6 in our study, that were identical to those previously designated Koblavi B5 or Popovic 5 and Popovic 6a or Tamayo B21a, respectively. Ribotype M5 was isolated between 1991 and 1993. This ribotype had a low level of genetic variation as detected by pulsed-field gel electrophoresis (PFGE). Ribotype M6 persisted from 1991 to 2000. However, PFGE profiles suggested that two epidemiologically unrelated strains coexisted within this single ribotype from 1995 until the end of the epidemic. We identified three new BglI ribotypes, Mx1, Mx2, and Mx3, from nontoxigenic V. cholerae O1 strains isolated between 1998 and 2000; one of them grouped strains positive for the toxin-coregulated pilus island. They differed from nontoxigenic clones isolated in Latin America and on the U.S. Gulf Coast and are probably autochthonous Mexican V. cholerae O1 variants. Most of these new variants were isolated from states surrounding the Gulf of Mexico, where the highest incidence of cholera in the country was recorded. Thus, the Mexican Gulf Coast, like the U.S. Gulf Coast, may act as an environmental reservoir of V. cholerae O1. PMID:19213700

  6. A Bistable Switch and Anatomical Site Control Vibrio cholerae Virulence Gene Expression in the Intestine

    DEFF Research Database (Denmark)

    Nielsen, Alex Toftgaard; Dolganov, N. A.; Rasmussen, Thomas

    2010-01-01

    A fundamental, but unanswered question in host-pathogen interactions is the timing, localization and population distribution of virulence gene expression during infection. Here, microarray and in situ single cell expression methods were used to study Vibrio cholerae growth and virulence gene...... expression during infection of the rabbit ligated ileal loop model of cholera. Genes encoding the toxin-coregulated pilus (TCP) and cholera toxin (CT) were powerfully expressed early in the infectious process in bacteria adjacent to epithelial surfaces. Increased growth was found to co......, a chemical inducer of virulence gene expression. Striking bifurcation of the population occurred during entry into stationary phase: one subpopulation continued to express tcpA, whereas the expression declined in the other subpopulation. ctxA, encoding the A subunit of CT, and toxT, encoding the proximal...

  7. Cholera after the consumption of raw oysters. A case report.

    Science.gov (United States)

    Klontz, K C; Tauxe, R V; Cook, W L; Riley, W H; Wachsmuth, I K

    1987-12-01

    In August 1986, a 76-year-old woman in Miami, Florida, developed profuse watery diarrhea and abdominal cramps. Two and four days before the onset of her illness, she had eaten six raw oysters at each of two restaurants in Miami. A stool specimen yielded toxigenic Vibrio cholerae O1 biotype El Tor, serotype Inaba. The results of toxin gene probing of the organism recovered from the patient differed significantly from those of other V. cholerae O1 isolates from the Gulf Coast and elsewhere in the world. A program of active surveillance identified no other cases of cholera in Miami. The source of the raw oysters eaten by the patient was traced to Louisiana. Her case represents the first reported case of cholera associated with eating raw oysters.

  8. Influence of gamma radiation on the immunobiological and immunochemical properties of cholera exotoxin

    International Nuclear Information System (INIS)

    Nedugova, G.I.; Rubtsov, I.V.; Samojlenko, I.I.

    1984-01-01

    Native cholera exotoxin (abacterial centrifugalized deposit) has been irradiated using gamma-installations with a 60 Co source. A high inactivating effect of gamma-radiation on native cholera exotoxin is established: with the increase of radiation dose cholerogenity decreased for certain (at the dose 50 kGy) a complete inactivation of all studied series of liquid filtrate-toxin took place), activity of permeability factor and toxicity for mice decreased. A higher radiostability of dry toxin preparations as compared with the liquid ones is detected. Sterilization effect of radiation is achieved at the dose 20 kGy for liquid preparations and at the dose of 30 kGy for dry ones. When preserving the irradiated preparations of raw toxin in different temperature regimes for 6 months to 1.5 year (observation time) toxic properties are not restored, immunogenous properties do not change

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

    OpenAIRE

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

    2014-01-01

    Vibrio cholerae classical (CL) biotype was isolated, along with biotype El Tor (ET) and altered ET carrying the cholera toxin (CTX) gene of CL biotype, during the 1991 cholera epidemic in Mexico, subsequently becoming endemic until 1997. Microbiological, molecular, and phylogenetic analyses of V. cholerae isolated from both clinical and environmental samples during 1998–2008 confirm important genetic events, namely predominance of ET over CL and altered ET in Mexico. Although altered ET is pr...

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

    Science.gov (United States)

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

    2014-01-01

    The seventh cholera pandemic caused by Vibrio cholerae O1 El Tor (ET) has been superseded in Asia and Africa by altered ET possessing the cholera toxin (CTX) gene of classical (CL) biotype. The CL biotype of V. cholerae was isolated, along with prototypic and altered ET, during the 1991 cholera epidemic in Mexico and subsequently remained endemic until 1997. Microbiological, molecular, and phylogenetic analyses of clinical and environmental V. cholerae isolated in Mexico between 1998 and 2008 revealed important genetic events favoring predominance of ET over CL and altered ET. V. cholerae altered ET was predominant after 1991 but not after 2000. V. cholerae strains isolated between 2001 and 2003 and a majority isolated in 2004 lacked CTX prophage (Φ) genes encoding CTX subunits A and B and repeat sequence transcriptional regulators of ET and CL biotypes: i.e., CTXΦ−. Most CTXΦ− V. cholerae isolated in Mexico between 2001 and 2003 also lacked toxin coregulated pili tcpA whereas some carried either tcpAET or a variant tcpA with noticeable sequence dissimilarity from tcpACL. The tcpA variants were not detected in 2005 after CTXΦ+ ET became dominant. All clinical and environmental V. cholerae O1 strains isolated during 2005–2008 in Mexico were CTXΦ+ ET, carrying an additional truncated CTXΦ instead of RS1 satellite phage. Despite V. cholerae CTXΦ− ET exhibiting heterogeneity in pulsed-field gel electrophoresis patterns, CTXΦ+ ET isolated during 2004–2008 displayed homogeneity and clonal relationship with V. cholerae ET N16961 and V. cholerae ET isolated in Peru. PMID:24958870

  11. The repertoire of glycosphingolipids recognized by Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    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.

  12. Safety of the recombinant cholera toxin B subunit, killed whole-cell (rBS-WC oral cholera vaccine in pregnancy.

    Directory of Open Access Journals (Sweden)

    Ramadhan Hashim

    Full Text Available Mass vaccinations are a main strategy in the deployment of oral cholera vaccines. Campaigns avoid giving vaccine to pregnant women because of the absence of safety data of the killed whole-cell oral cholera (rBS-WC vaccine. Balancing this concern is the known higher risk of cholera and of complications of pregnancy should cholera occur in these women, as well as the lack of expected adverse events from a killed oral bacterial vaccine.From January to February 2009, a mass rBS-WC vaccination campaign of persons over two years of age was conducted in an urban and a rural area (population 51,151 in Zanzibar. Pregnant women were advised not to participate in the campaign. More than nine months after the last dose of the vaccine was administered, we visited all women between 15 and 50 years of age living in the study area. The outcome of pregnancies that were inadvertently exposed to at least one oral cholera vaccine dose and those that were not exposed was evaluated. 13,736 (94% of the target women in the study site were interviewed. 1,151 (79% of the 1,453 deliveries in 2009 occurred during the period when foetal exposure to the vaccine could have occurred. 955 (83% out of these 1,151 mothers had not been vaccinated; the remaining 196 (17% mothers had received at least one dose of the oral cholera vaccine. There were no statistically significant differences in the odds ratios for birth outcomes among the exposed and unexposed pregnancies.We found no statistically significant evidence of a harmful effect of gestational exposure to the rBS-WC vaccine. These findings, along with the absence of a rational basis for expecting a risk from this killed oral bacterial vaccine, are reassuring but the study had insufficient power to detect infrequent events.ClinicalTrials.gov NCT00709410.

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

    International Nuclear Information System (INIS)

    Das, G.; Das, J.

    1983-01-01

    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. (orig.)

  14. Experimental parameters differentially affect the humoral response of the cholera-toxin-based murine model of food allergy

    DEFF Research Database (Denmark)

    Kroghsbo, S.; Christensen, Hanne Risager; Frøkiær, Hanne

    2003-01-01

    Background: Recent studies have developed a murine model of IgE-mediated food allergy based on oral coadministration of antigen and cholera toxin (CT) to establish a maximal response for studying immunopathogenic mechanisms and immunotherapeutic strategies. However, for studying subtle...... interested in characterizing the individual effects of the parameters in the CT-based model: CT dose, antigen type and dose, and number of immunizations. Methods: BALB/c mice were orally sensitized weekly for 3 or 7 weeks with graded doses of CT and various food antigens (soy-trypsin inhibitor, ovalbumin...... of the antibody response depended on the type of antigen and number of immunizations. Conclusions: The critical parameters of the CT-based murine allergy model differentially control the intensity and kinetics of the developing immune response. Adjustment of these parameters could be a key tool for tailoring...

  15. 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.

  16. Lipopolysaccharide-specific memory B cell responses to an attenuated live cholera vaccine are associated with protection against Vibrio cholerae infection.

    Science.gov (United States)

    Haney, Douglas J; Lock, Michael D; Gurwith, Marc; Simon, Jakub K; Ishioka, Glenn; Cohen, Mitchell B; Kirkpatrick, Beth D; Lyon, Caroline E; Chen, Wilbur H; Sztein, Marcelo B; Levine, Myron M; Harris, Jason B

    2018-05-11

    The single-dose live attenuated vaccine CVD 103-HgR protects against experimental Vibrio cholerae infection in cholera-naïve adults for at least 6 months after vaccination. While vaccine-induced vibriocidal seroconversion is associated with protection, vibriocidal titers decline rapidly from their peak 1-2 weeks after vaccination. Although vaccine-induced memory B cells (MBCs) might mediate sustained protection in individuals without detectable circulating antibodies, it is unknown whether oral cholera vaccination induces a MBC response. In a study that enrolled North American adults, we measured lipopolysaccharide (LPS)- and cholera toxin (CtxB)-specific MBC responses to PXVX0200 (derived from the CVD 103-HgR strain) and assessed stool volumes following experimental Vibrio cholerae infection. We then evaluated the association between vaccine-induced MBC responses and protection against cholera. There was a significant increase in % CT-specific IgG, % LPS-specific IgG, and % LPS-specific IgA MBCs which persisted 180 days after vaccination as well as a significant association between vaccine-induced increase in % LPS-specific IgA MBCs and lower post-challenge stool volume (r = -0.56, p < 0.001). Oral cholera vaccination induces antigen-specific MBC responses, and the anamnestic LPS-specific responses may contribute to long-term protection and provide correlates of the duration of vaccine-induced protection. NCT01895855. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  17. Expression of cholera toxin B–proinsulin fusion protein in lettuce and tobacco chloroplasts – oral administration protects against development of insulitis in non-obese diabetic mice

    OpenAIRE

    Ruhlman, Tracey; Ahangari, Raheleh; Devine, Andrew; Samsam, Mohtahsem; Daniell, Henry

    2007-01-01

    Lettuce and tobacco chloroplast transgenic lines expressing the cholera toxin B subunit–human proinsulin (CTB-Pins) fusion protein were generated. CTB-Pins accumulated up to ~16% of total soluble protein (TSP) in tobacco and up to ~2.5% of TSP in lettuce. Eight milligrams of powdered tobacco leaf material expressing CTB-Pins or, as negative controls, CTB–green fluorescent protein (CTB-GFP) or interferon–GFP (IFN-GFP), or untransformed leaf, were administered orally, each week for 7 weeks, to ...

  18. Generation of transgenic corn-derived Actinobacillus pleuropneumoniae ApxIIA fused with the cholera toxin B subunit as a vaccine candidate

    Science.gov (United States)

    Shin, Min-Kyoung; Jung, Myung Hwan; Lee, Won-Jung; Choi, Pil Son; Jang, Yong-Suk

    2011-01-01

    Corn, one of the most important forage crops worldwide, has proven to be a useful expression vehicle due to the availability of established transformation procedures for this well-studied plant. The exotoxin Apx, a major virulence factor, is recognized as a common antigen of Actinobacillus (A.) pleuropneumoniae, the causative agent of porcine pleuropneumonia. In this study, a cholera toxin B (CTB)-ApxIIA#5 fusion protein and full-size ApxIIA expressed in corn seed, as a subunit vaccine candidate, were observed to induce Apx-specific immune responses in mice. These results suggest that transgenic corn-derived ApxIIA and CTB-ApxIIA#5 proteins are potential vaccine candidates against A. pleuropneumoniae infection. PMID:22122907

  19. Novel mucosal DNA-MVA HIV vaccination in which DNA-IL-12 plus Cholera Toxin B subunit (CTB) cooperates to enhance cellular systemic and mucosal genital tract immunity

    OpenAIRE

    Maeto, Cynthia Alejandra; Rodríguez, Ana María; Holgado, María Pía; Falivene, Juliana; Gherardi, Maria Magdalena

    2017-01-01

    Induction of local antiviral immune responses at the mucosal portal surfaces where HIV-1 and other viral pathogens are usually first encountered remains a primary goal for most vaccines against mucosally acquired viral infections. Exploring mucosal immunization regimes in order to find optimal vector combinations and also appropriate mucosal adjuvants in the HIV vaccine development is decisive. In this study we analyzed the interaction of DNA-IL-12 and cholera toxin B subunit (CTB) after thei...

  20. Dorsal lateral geniculate substructure in the Long-Evans rat: A cholera toxin B-subunit study

    Directory of Open Access Journals (Sweden)

    Claire B. Discenza

    2012-09-01

    Full Text Available The pigmented rat is an increasingly important model in visual neuroscience research, yet the lamination of retinal projections in the dLGN has not been examined in sufficient detail. From previous studies it was known that most of the rat dLGN receives monocular input from the contralateral eye, with a small island receiving predominantly ipsilateral projections. Here we revisit the question using cholera toxin B subunit (CTB, a tracer that efficiently fills retinal terminals after intra-ocular injection. We imaged retinal termini throughout the dLGN at 0.5 um resolution and traced areas of ipsilateral and contralateral terminals to obtain a high resolution 3D reconstruction of the projection pattern. Retinal termini in the dLGN are well segregated by eye of origin, as expected. We find, however, that the ipsilateral projections form multiple discrete projection zones in three dimensions, not the single island previously described. It remains to be determined whether these subdomains represent distinct functional sublaminae, as is the case in other mammals.

  1. Intranasal delivery of cholera toxin induces th17-dominated T-cell response to bystander antigens.

    Directory of Open Access Journals (Sweden)

    Jee-Boong Lee

    Full Text Available Cholera toxin (CT is a potent vaccine adjuvant, which promotes mucosal immunity to protein antigen given by nasal route. It has been suggested that CT promotes T helper type 2 (Th2 response and suppresses Th1 response. We here report the induction of Th17-dominated responses in mice by intranasal delivery of CT. This dramatic Th17-driving effect of CT, which was dependent on the B subunit, was observed even in Th1 or Th2-favored conditions of respiratory virus infection. These dominating Th17 responses resulted in the significant neutrophil accumulation in the lungs of mice given CT. Both in vitro and in vivo treatment of CT induced strongly augmented IL-6 production, and Th17-driving ability of CT was completely abolished in IL-6 knockout mice, indicating a role of this cytokine in the Th17-dominated T-cell responses by CT. These data demonstrate a novel Th17-driving activity of CT, and help understand the mechanisms of CT adjuvanticity to demarcate T helper responses.

  2. Vibrio cholerae Colonization of Soft-Shelled Turtles.

    Science.gov (United States)

    Wang, Jiazheng; Yan, Meiying; Gao, He; Lu, Xin; Kan, Biao

    2017-07-15

    Vibrio cholerae is an important human pathogen and environmental microflora species that can both propagate in the human intestine and proliferate in zooplankton and aquatic organisms. Cholera is transmitted through food and water. In recent years, outbreaks caused by V. cholerae -contaminated soft-shelled turtles, contaminated mainly with toxigenic serogroup O139, have been frequently reported, posing a new foodborne disease public health problem. In this study, the colonization by toxigenic V. cholerae on the body surfaces and intestines of soft-shelled turtles was explored. Preferred colonization sites on the turtle body surfaces, mainly the carapace and calipash of the dorsal side, were observed for the O139 and O1 strains. Intestinal colonization was also found. The colonization factors of V. cholerae played different roles in the colonization of the soft-shelled turtle's body surface and intestine. Mannose-sensitive hemagglutinin (MSHA) of V. cholerae was necessary for body surface colonization, but no roles were found for toxin-coregulated pili (TCP) or N -acetylglucosamine-binding protein A (GBPA). Both TCP and GBPA play important roles for colonization in the intestine, whereas the deletion of MSHA revealed only a minor colonization-promoting role for this factor. Our study demonstrated that V. cholerae can colonize the surfaces and the intestines of soft-shelled turtles and indicated that the soft-shelled turtles played a role in the transmission of cholera. In addition, this study showed that the soft-shelled turtle has potential value as an animal model in studies of the colonization and environmental adaption mechanisms of V. cholerae in aquatic organisms. IMPORTANCE Cholera is transmitted through water and food. Soft-shelled turtles contaminated with Vibrio cholerae (commonly the serogroup O139 strains) have caused many foodborne infections and outbreaks in recent years, and they have become a foodborne disease problem. Except for epidemiological

  3. Effects of cholera toxin on human colon carcinoma cell lines.

    Science.gov (United States)

    Barkla, D H; Whitehead, R H; Hayward, I P

    1992-10-01

    This study reports on changes in morphology and membrane transport in 5 human colon carcinoma cell lines treated with cholera toxin (CT). Three of the cell lines that grew as monolayers (LIM 1215, LIM 1899, LIM 2099) and 1 that grew as floating clumps (LIM 2408) did not show morphological changes after CT treatment. However, cell line LIM 1863 that grows as floating "crypt-like" organoids showed rapid and distinctive changes in morphology and membrane transport after CT treatment. At 1 and 6 hrs after CT treatment, light and transmission electron microscopy revealed rapid dilatation of the central lumen of organoids and the appearance of 2 populations of apical vesicular inclusions. The first population was unusual in being non-membrane bound and limited by fuzzy filamentous material. The second population was membrane bound. Scanning electron microscopy at 1-6 hr after CT treatment showed swelling and loss of surface microvilli on some, but not all, cells. At 24 hr after CT treatment the majority of organoids showed evidence of fluid accumulation and small apical vesicles coalesced to form large single vacuoles that obliterated normal cell morphology. By 48 hr, continued swelling produced extreme attenuation of the plasma membrane with cells taking on an "endothelial cell-like" appearance. The response to CT was dose-dependent. Uptake studies using 86Rubidium and blocking studies using ouabain and amiloride indicated that CT is acting on the Na+/K+ ATPase membrane pump to cause the increased fluid uptake by LIM 1863 cells. This study is the first to report specific morphological changes in intestine-derived cells in response to CT.(ABSTRACT TRUNCATED AT 250 WORDS)

  4. Influence of gamma radiation on the immunological and immunochemical properties of cholera

    International Nuclear Information System (INIS)

    Nedugova, G.I.; Rubtsov, I.V.; Samojlenko, I.I.

    1984-01-01

    Results of studying the effect of gamma-radiation on immunochemical properties and serologic activity of unpurified cholera exotoxin are presented. It is found that in irradiated toxin preparations physico-chemical alterations take place as the dose of ionizing radiation increases, which brings about the increase in electrophoretic mobility, aggregation of protein components. It is shown that serologic activity contained in antigene toxin preparations retains within the limits of radiation doses studied

  5. Comparative genomic characterization of a Thailand-Myanmar isolate, MS6, of Vibrio cholerae O1 El Tor, which is phylogenetically related to a "US Gulf Coast" clone.

    Science.gov (United States)

    Okada, Kazuhisa; Na-Ubol, Mathukorn; Natakuathung, Wirongrong; Roobthaisong, Amonrattana; Maruyama, Fumito; Nakagawa, Ichiro; Chantaroj, Siriporn; Hamada, Shigeyuki

    2014-01-01

    The cholera outbreaks in Thailand during 2007-2010 were exclusively caused by the Vibrio cholerae O1 El Tor variant carrying the cholera toxin gene of the classical biotype. We previously isolated a V. cholerae O1 El Tor strain from a patient with diarrhea and designated it MS6. Multilocus sequence-typing analysis revealed that MS6 is most closely related to the U. S. Gulf Coast clone with the exception of two novel housekeeping genes. The nucleotide sequence of the genome of MS6 was determined and compared with those of 26 V. cholerae strains isolated from clinical and environmental sources worldwide. We show here that the MS6 isolate is distantly related to the ongoing seventh pandemic V. cholerae O1 El Tor strains. These strains differ with respect to polymorphisms in housekeeping genes, seventh pandemic group-specific markers, CTX phages, two genes encoding predicted transmembrane proteins, the presence of metY (MS6_A0927) or hchA/luxR in a highly conserved region of the V. cholerae O1 serogroup, and a superintegron (SI). We found that V. cholerae species carry either hchA/luxR or metY and that the V. cholerae O1 clade commonly possesses hchA/luxR, except for MS6 and U. S. Gulf Coast strains. These findings illuminate the evolutionary relationships among V. cholerae O1 strains. Moreover, the MS6 SI carries a quinolone-resistance gene cassette, which was closely related with those present in plasmid-borne integrons of other gram-negative bacteria. Phylogenetic analysis reveals that MS6 is most closely related to a U. S. Gulf Coast clone, indicating their divergence before that of the El Tor biotype strains from a common V. cholerae O1 ancestor. We propose that MS6 serves as an environmental aquatic reservoir of V. cholerae O1.

  6. 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

  7. The increased severity in patients presenting to hospital with diarrhea in Dhaka, Bangladesh since the emergence of the hybrid strain of Vibrio cholerae O1 is not unique to cholera patients.

    Science.gov (United States)

    Chowdhury, Fahima; Kuchta, Alison; Khan, Ashraful Islam; Faruque, A S G; Calderwood, Stephen B; Ryan, Edward T; Qadri, Firdausi

    2015-11-01

    A hybrid strain of Vibrio cholerae O1 El Tor that expresses a classical cholera toxin (CT) emerged in 2001. This hybrid variant rapidly replaced the previous El Tor strain around the world. The global emergence of this variant coincided with anecdotal reports that cholera patients were presenting with more severe dehydration and disease in many locations. A comparison was made of the severity of disease before and after the emergence of the hybrid strain in cholera patients attending an icddr,b hospital in Dhaka, Bangladesh. It was found that cholera patients presented with more severe dehydration and severe disease in the later period. However, this was also true for all non-cholera patients as well. In addition, in sub-analyses of patients who presented with rotavirus and enterotoxigenic Escherichia coli (ETEC), similar results were found. Comparing the two periods for differences in patient characteristics, nutritional status, vaccination status, and income, no plausible cause for patients presenting with more severe disease was identified in the later period. As a shift in severity for both cholera and non-cholera was observed, these results indicate that the altered El Tor strain cannot fully explain the difference in cholera severity before and after 2001. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  8. Co-administration of cholera toxin and apple polyphenol extract as a novel and safe mucosal adjuvant strategy.

    Science.gov (United States)

    Yoshino, Naoto; Fujihashi, Kohtaro; Hagiwara, Yukari; Kanno, Hiroyuki; Takahashi, Kiyomi; Kobayashi, Ryoki; Inaba, Noriyuki; Noda, Masatoshi; Sato, Shigehiro

    2009-07-30

    Although native cholera toxin (CT) is an extremely effective adjuvant, its toxicity prevents its use in humans. We report here that apple polyphenol extract (APE), obtained from unripe apples, reduces CT-induced morphological changes and cAMP accumulation. Based upon this finding, we have attempted to design a novel, effective and safe mucosal vaccine by using CT with several dosages of APE as nasal adjuvants. Mice nasally immunized with OVA plus CT and an optimal dosage of APE showed significantly reduced levels of inflammatory responses as well as total and OVA-specific IgE antibodies when compared with mice given without APE. However, levels of both mucosal and systemic OVA-specific antibody responses were maintained. Further, APE significantly down-regulated accumulation of CT in the olfactory nerves and epithelium. In summary, an optimal dosage of APE would take full advantage of mucosal adjuvanticity of native CT without any toxicity for application in humans.

  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......, all originating from shrimp produced in aquaculture. Six isolated strains agglutinated in polyvalent O antisera, but did not agglutinate in Ogawa or Inaba antisera. The six strains were resistant to colistin and sulfisoxazole; three strains also showed resistance to ampicillin. None of the strains...... contained plasmids or genes encoding cholera toxin (CT) or heat-stable enterotoxin (NAG-ST), The absence of V. cholerae O1 and the low number of samples containing CT and NAG-ST negative non-Ol strains in imported shrimp suggest that I! cholerae in such products may not constitute a public health problem....

  10. Cholera between 1991 and 1997 in Mexico Was Associated with Infection by Classical, El Tor, and El Tor Variants of Vibrio cholerae▿

    OpenAIRE

    Alam, Munirul; Nusrin, Suraia; Islam, Atiqul; Bhuiyan, Nurul A.; Rahim, Niaz; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Gil, Ana I.; Watanabe, Haruo; Morita, Masatomo; Nair, G. Balakrish; Cravioto, Alejandro

    2010-01-01

    Vibrio cholerae O1 biotype El Tor (ET), the cause of the current 7th pandemic, has recently been replaced in Asia and Africa by an altered ET biotype possessing cholera toxin (CTX) of the classical (CL) biotype that originally caused the first six pandemics before becoming extinct in the 1980s. Until recently, the ET prototype was the biotype circulating in Peru; a detailed understanding of the evolutionary trend of V. cholerae causing endemic cholera in Latin America is lacking. The present ...

  11. Steady-state levels of G-protein beta-subunit expression are regulated by treatment of cells with bacterial toxins

    International Nuclear Information System (INIS)

    Watkins, D.C.; Northup, J.K.; Malbon, C.C.

    1987-01-01

    Cultures of 3T3-L1 cells were incubated with either 10 ng/ml cholera toxin or 10 ng/ml pertussis toxin from 4 days prior to the initiation of differentiation and throughout the subsequent incubation. Toxin concentrations were sufficient to completely prevent the labelling of alpha-subunits with [ 32 P]NAD + and pertussis toxin and to prevent by more than 90% the labelling with [ 32 P]NAD + and cholera toxin in membranes prepared from these cells. Neither toxin prevented the differentiation to the adipocyte phenotype. Neither toxin prevented the increases in the relative amounts of G-proteins which occur upon differentiation. Both toxins dramatically decreased the amount of beta-subunits. As measured by quantitative immunoblotting with antisera specific for both the 35 kDa and 36 kDa beta-subunits, levels of beta-subunit were decreased by more than 50% of steady-state level of control cells. Thus, bacterial toxins which modifies G-protein alpha-subunits are capable of modulating the levels of beta-subunits in vivo. The basis for the regulation of G-protein subunit expression by bacterial toxins is under study

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

    Science.gov (United States)

    Madhusudana, Rao B; Surendran, P K

    2013-06-01

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

  13. Phenotypic and genetic characterization of Vibrio cholerae O1 clinical isolates collected through national antimicrobial resistance surveillance network in Nepal.

    Science.gov (United States)

    Shakya, Geeta; Kim, Dong Wook; Clemens, John D; Malla, Sarala; Upadhyaya, Bishnu Prasad; Dumre, Shyam Prakash; Shrestha, Sirjana Devi; Adhikari, Shailaja; Sharma, Supriya; Rijal, Nisha; Shrestha, Sanjaya K; Mason, Carl; Kansakar, Palpasa

    2012-08-01

    Cholera occurs in sporadic cases and outbreaks in Nepal each year. Vibrio cholerae O1 (n = 522) isolated during 2007-2010 from diarrheal patients at 10 different hospital laboratories in Nepal were characterized. Biochemical and serologic identifications showed that all the isolates belonged to serogroup O1, El Tor biotype. Except 72 isolates of Inaba serotype isolated in the year 2007, all the remaining isolates were of Ogawa serotype. All isolates were resistant to nalidixic acid and furazolidone. Resistance to tetracycline, ciprofloxacin, erythromycin and co-trimoxazole were 21, 4, 16 and 90 % respectively. Seventy-seven of these isolates were selected for further characterization for ctxB gene and MLVA typing. Two different variants of classical type cholera toxin were observed. Ogawa strains from 2007 and 2010-Western Nepal outbreak harbored CTX-3 type cholera toxin, whereas Inaba serotypes in 2007 and the remaining Ogawa serotypes in 2008-2010 harbored CTX 3b-type toxin. MLVA analysis showed circulation of four different groups of altered V. cholerae O1 El Tor strains. Two different profiles were seen among 2007 Inaba (9, 3, 6, x, x) and Ogawa (10, 7, 6, x, x) isolates. The MLVA profile of 2008 and 2009 Ogawa isolates were similar to those of Inaba strains of 2007. Isolates from 2010 also showed three different MLVA profiles; profile 9, 3, 6, x, x in 3 isolates, 11, 7, 6, x, x among 2010 Western Nepal outbreak strains and profile 8, 3, 6, x, x among isolates from Butwal and Kathmandu.

  14. Cholera between 1991 and 1997 in Mexico Was Associated with Infection by Classical, El Tor, and El Tor Variants of Vibrio cholerae▿

    Science.gov (United States)

    Alam, Munirul; Nusrin, Suraia; Islam, Atiqul; Bhuiyan, Nurul A.; Rahim, Niaz; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Gil, Ana I.; Watanabe, Haruo; Morita, Masatomo; Nair, G. Balakrish; Cravioto, Alejandro

    2010-01-01

    Vibrio cholerae O1 biotype El Tor (ET), the cause of the current 7th pandemic, has recently been replaced in Asia and Africa by an altered ET biotype possessing cholera toxin (CTX) of the classical (CL) biotype that originally caused the first six pandemics before becoming extinct in the 1980s. Until recently, the ET prototype was the biotype circulating in Peru; a detailed understanding of the evolutionary trend of V. cholerae causing endemic cholera in Latin America is lacking. The present retrospective microbiological, molecular, and phylogenetic study of V. cholerae isolates recovered in Mexico (n = 91; 1983 to 1997) shows the existence of the pre-1991 CL biotype and the ET and CL biotypes together with the altered ET biotype in both epidemic and endemic cholera between 1991 and 1997. According to sero- and biotyping data, the altered ET, which has shown predominance in Mexico since 1991, emerged locally from ET and CL progenitors that were found coexisting until 1997. In Latin America, ET and CL variants shared a variable number of phenotypic markers, while the altered ET strains had genes encoding the CL CTX (CTXCL) prophage, ctxBCL and rstRCL, in addition to resident rstRET, as the underlying regional signature. The distinct regional fingerprints for ET in Mexico and Peru and their divergence from ET in Asia and Africa, as confirmed by subclustering patterns in a pulsed-field gel electrophoresis (NotI)-based dendrogram, suggest that the Mexico epidemic in 1991 may have been a local event and not an extension of the epidemics occurring in Asia and South America. Finally, the CL biotype reservoir in Mexico is unprecedented and must have contributed to the changing epidemiology of global cholera in ways that need to be understood. PMID:20668130

  15. Case studies in cholera: lessons in medical history and science.

    Science.gov (United States)

    Kavic, S. M.; Frehm, E. J.; Segal, A. S.

    1999-01-01

    Cholera, a prototypical secretory diarrheal disease, is an ancient scourge that has both wrought great suffering and taught many valuable lessons, from basic sanitation to molecular signal transduction. Victims experience the voluminous loss of bicarbonate-rich isotonic saline at a rate that may lead to hypovolemic shock, metabolic acidosis, and death within afew hours. Intravenous solution therapy as we know it was first developed in an attempt to provide life-saving volume replacement for cholera patients. Breakthroughs in epithelial membrane transport physiology, such as the discovery of sugar and salt cotransport, have paved the way for oral replacement therapy in areas of the world where intravenous replacement is not readily available. In addition, the discovery of the cholera toxin has yielded vital information about toxigenic infectious diseases, providing a framework in which to study fundamental elements of intracellular signal transduction pathways, such as G-proteins. Cholera may even shed light on the evolution and pathophysiology of cystic fibrosis, the most commonly inherited disease among Caucasians. The goal of this paper is to review, using case studies, some of the lessons learned from cholera throughout the ages, acknowledging those pioneers whose seminal work led to our understanding of many basic concepts in medical epidemiology, microbiology, physiology, and therapeutics. PMID:11138935

  16. Effect of High Pressure and Heat on Bacterial Toxins

    Directory of Open Access Journals (Sweden)

    Dirk Margosch

    2005-01-01

    Full Text Available Even though the inactivation of microorganisms by high pressure treatment is a subject of intense investigations, the effect of high pressure on bacterial toxins has not been studied so far. In this study, the influence of combined pressure/temperature treatment (0.1 to 800 MPa and 5 to 121 °C on bacterial enterotoxins was determined. Therefore, heat-stable enterotoxin (STa of cholera toxin (CT from Vibrio cholerae, staphylococcal enterotoxins A-E, haemolysin BL (HBL from Bacillus cereus, and Escherichia coli (STa were subjected to different treatment schemes. Structural alterations were monitored in enzyme immunoassays (EIAs. Cytotoxicity of the pressure treated supernatant of toxigenic B. cereus DSM 4384 was investigated with Vero cells. High pressure of 200 to 800 MPa at 5 °C leads to a slight increase of the reactivity of the STa of E. coli. However, reactivity decreased at 800 MPa and 80 °C to (66±21 % after 30 min and to (44±0.3 % after 128 min. At ambient pressure no decrease in EIA reactivity could be observed after 128 min. Pressurization (0.1 to 800 MPa of heat stable monomeric staphylococcal toxins at 5 and 20 °C showed no effect. A combined heat (80 °C and pressure (0.1 to 800 MPa treatment lead to a decrease in the immuno-reactivity to 20 % of its maximum. For cholera toxin a significant loss in latex agglutination was observable only at 80 °C and 800 MPa for holding times higher than 20 min. Interestingly, the immuno-reactivity of B. cereus HBL toxin increased with the increase of pressure (182 % at 800 MPa, 30 °C, and high pressure showed only minor effects on cytotoxicity to Vero cells. Our results indicate that pressurization can increase inactivation observed by heat treatment, and combined treatments may be effective at lower temperatures and/or shorter incubation time.

  17. The three-dimensional crystal structure of cholera toxin

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Rong-Guang; Westbrook, M.L.; Nance, S.; Spangler, B.D. [Argonne National Lab., IL (United States); Scott, D.L. [Yale Univ., New Haven, CT (United States). Dept. of Molecular Biophysics and Biochemistry; Westbrook, E.M. [Northwestern Univ., Evanston, IL (United States)

    1996-02-01

    The clinical manifestations of cholera are largely attributable to the actions of a secreted hexameric AB{sub 5} enterotoxin (choleragen). We have solved the three-dimensional structure of choleragen at 2.5 {Angstrom} resolution and compared the refined coordinates with those of choleragenoid (isolated B pentamer) and the heat-labile enterotoxin from Escherichia coli (LT). The crystalline coordinates provide a detailed view of the stereochemistry implicated in binding to GM1 gangliosides and in carrying out ADP-ribosylation. The A2 chain of choleragen, in contrast to that of LT, is a nearly continuous {alpha}-helix with an interpretable carboxyl tail.

  18. Analysis of a cholera toxin B subunit (CTB) and human mucin 1 (MUC1) conjugate protein in a MUC1-tolerant mouse model.

    Science.gov (United States)

    Pinkhasov, Julia; Alvarez, M Lucrecia; Pathangey, Latha B; Tinder, Teresa L; Mason, Hugh S; Walmsley, Amanda M; Gendler, Sandra J; Mukherjee, Pinku

    2010-12-01

    Since epithelial mucin 1 (MUC1) is associated with several adenocarcinomas at the mucosal sites, it is pertinent to test the efficacy of a mucosally targeted vaccine formulation. The B subunit of the Vibrio cholerae cholera toxin (CTB) has great potential to act as a mucosal carrier for subunit vaccines. In the present study we evaluated whether a MUC1 tandem repeat (TR) peptide chemically linked to CTB would break self-antigen tolerance in the transgenic MUC1-tolerant mouse model (MUC1.Tg) through oral or parenteral immunizations. We report that oral immunization with the CTB-MUC1 conjugate along with mucosal adjuvant, unmethylated CpG oligodeoxynucleotide (ODN) and interleukin-12 (IL-12) did not break self-antigen tolerance in MUC1.Tg mice, but induced a strong humoral response in wild-type C57BL/6 mice. However, self-antigen tolerance in the MUC1.Tg mouse model was broken after parenteral immunizations with different doses of the CTB-MUC1 conjugate protein and with the adjuvant CpG ODN co-delivered with CTB-MUC1. Importantly, mice immunized systemically with CpG ODN alone and with CTB-MUC1 exhibited decreased tumor burden when challenged with a mammary gland tumor cell line that expresses human MUC1.

  19. Analysis of a Cholera Toxin B Subunit (CTB) and Human Mucin 1 (MUC1) Conjugate Protein in a MUC1 Tolerant Mouse Model

    Science.gov (United States)

    Pinkhasov, Julia; Alvarez, M. Lucrecia; Pathangey, Latha B.; Tinder, Teresa L.; Mason, Hugh S.; Walmsley, Amanda M.; Gendler, Sandra J.; Mukherjee, Pinku

    2011-01-01

    Since epithelial mucin 1 (MUC1) is associated with several adenocarcinomas at mucosal sites, it is pertinent to test the efficacy of a mucosally targeted vaccine formulation. The B subunit of the Vibrio cholerae cholera toxin (CTB) has great potential to act as a mucosal carrier for subunit vaccines. In the present study we evaluated whether a MUC1 tandem repeat (TR) peptide chemically linked to CTB would break self-antigen tolerance in the transgenic MUC1 tolerant mouse model (MUC1.Tg) through oral or parenteral immunizations. We report that oral immunization with the CTB-MUC1 conjugate along with mucosal adjuvant, unmethylated CpG oligodeoxynucleotide (ODN) and interleukin-12 (IL-12), did not break self-antigen tolerance in MUC1.Tg mice, but induced a strong humoral response in wild-type C57BL/6 mice. However, self-antigen tolerance in the MUC1.Tg mouse model was broken after parenteral immunizations with different doses of the CTB-MUC1 conjugate protein and with the adjuvant CpG ODN co-delivered with CTB-MUC1. Importantly, mice immunized systemically with CpG ODN alone and with CTB-MUC1 exhibited decreased tumor burden when challenged with a mammary gland tumor cell line that expresses human MUC1. PMID:20824430

  20. Histochemical detection of GM1 ganglioside using cholera toxin-B subunit. Evaluation of critical factors optimal for in situ detection with special emphasis to acetone pre-extraction

    Science.gov (United States)

    Petr, T.; Šmíd, V.; Šmídová, J.; Hůlková, H.; Jirkovská, M.; Elleder, M.; Muchová, L.; Vítek, L.; Šmíd, F.

    2010-01-01

    A comparison of histochemical detection of GM1 ganglioside in cryostat sections using cholera toxin B-subunit after fixation with 4% formaldehyde and dry acetone gave tissue-dependent results. In the liver no pre-treatment showed detectable differences related to GM1 reaction products, while studies in the brain showed the superiority of acetone pre-extraction (followed by formaldehyde), which yielded sharper images compared with the diffuse, blurred staining pattern associated with formaldehyde. Therefore, the aim of our study was to define the optimal conditions for the GM1 detection using cholera toxin B-subunit. Ganglioside extractability with acetone, the ever neglected topic, was tested comparing anhydrous acetone with acetone containing admixture of water. TLC analysis of acetone extractable GM1 ganglioside from liver sections did not exceed 2% of the total GM1 ganglioside content using anhydrous acetone at −20°C, and 4% at room temperature. The loss increased to 30.5% using 9:1 acetone/water. Similarly, photometric analysis of lipid sialic acid, extracted from dried liver homogenates with anhydrous acetone, showed the loss of gangliosides into acetone 3.0±0.3% only. The loss from dried brain homogenate was 9.5±1.1%. Thus, anhydrous conditions (dry tissue samples and anhydrous acetone) are crucial factors for optimal in situ ganglioside detection using acetone pre-treatment. This ensures effective physical fixation, especially in tissues rich in polar lipids (precipitation, prevention of in situ diffusion), and removal of cholesterol, which can act as a hydrophobic blocking barrier. PMID:20558344

  1. Histochemical detection of GM1 ganglioside using cholera toxin-B subunit. Evaluation of critical factors optimal for in situ detection with special emphasis to acetone pre-extraction

    Directory of Open Access Journals (Sweden)

    T. Petr

    2010-05-01

    Full Text Available A comparison of histochemical detection of GM1 ganglioside in cryostat sections using cholera toxin B-subunit after fixation with 4% formaldehyde and dry acetone gave tissue-dependent results. In the liver no pre-treatment showed detectable differences related to GM1 reaction products, while studies in the brain showed the superiority of acetone pre-extraction (followed by formaldehyde, which yielded sharper images compared with the diffuse, blurred staining pattern associated with formaldehyde. Therefore, the aim of our study was to define the optimal conditions for the GM1 detection using cholera toxin B-subunit. Ganglioside extractability with acetone, the ever neglected topic, was tested comparing anhydrous acetone with acetone containing admixture of water. TLC analysis of acetone extractable GM1 ganglioside from liver sections did not exceed 2% of the total GM1 ganglioside content using anhydrous acetone at -20°C, and 4% at room temperature. The loss increased to 30.5% using 9:1 acetone/water. Similarly, photometric analysis of lipid sialic acid, extracted from dried liver homogenates with anhydrous acetone, showed the loss of gangliosides into acetone 3.0±0.3% only. The loss from dried brain homogenate was 9.5±1.1%. Thus, anhydrous conditions (dry tissue samples and anhydrous acetone are crucial factors for optimal in situ ganglioside detection using acetone pre-treatment. This ensures effective physical fixation, especially in tissues rich in polar lipids (precipitation, prevention of in situ diffusion, and removal of cholesterol, which can act as a hydrophobic blocking barrier.

  2. Outbreaks of cholera-like diarrhoea caused by enterotoxigenic Escherichia coli in the Brazilian Amazon Rainforest.

    Science.gov (United States)

    Vicente, Ana C P; Teixeira, Luiz F M; Iniguez-Rojas, L; Luna, M G; Silva, L; Andrade, J R C; Guth, B E C

    2005-09-01

    The relationship between enteropathogens and severe diarrhoea in the Brazilian Amazon is poorly understood. In 1998, outbreaks of acute diarrhoea clinically diagnosed as cholera occurred in two small villages localized far from the main cholera route in the Brazilian rainforest. PCR was performed on some enteropathogens and heat-labile (LT) and/or heat-stable (STh) toxin genes, the virulence determinants of enterotoxigenic Escherichia coli (ETEC), were detected. Further characterization of ETEC isolates revealed the presence of two clones, one from each outbreak. One presenting serotype O167:H5 harboured LT-I and STh toxin genes and expressed the CS5CS6 colonization factor. The other, a non-typeable serotype, was positive for the LT-I gene and expressed the CS7 colonization factor. The current study demonstrates the importance of molecular diagnosis in regions such as the Amazon basin, where the enormous distances and local support conditions make standard laboratory diagnosis difficult. Here we also show that the mis-identified cholera cases were in fact associated with ETEC strains. This is the first report of ETEC, molecularly characterized as the aetiological agent of severe diarrhoea in children and adults in the Brazilian Amazon Rainforest.

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

    Directory of Open Access Journals (Sweden)

    M Shamim Hasan Zahid

    Full Text Available 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.

  4. 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.

  5. 3-Amino 1,8-naphthalimide, a structural analog of the anti-cholera drug virstatin inhibits chemically-biased swimming and swarming motility in vibrios

    Science.gov (United States)

    Wang, Hongxia; Silva, Anisia J.; Benitez, Jorge A.

    2017-01-01

    A screen for inhibitors of Vibrio cholerae motility identified the compound 3-amino 1,8-naphthalimide (3-A18NI), a structural analog of the cholera drug virstatin. Similar to virstatin, 3-A18NI diminished cholera toxin production. In contrast, 3-A18NI impeded swimming and/or swarming motility of V. cholerae and V. parahemolyticus suggesting that it could target the chemotaxis pathway shared by the polar and lateral flagellar system of vibrios. 3-A18NI did not inhibit the expression of V. cholerae major flagellin FlaA or the assembly of its polar flagellum. Finally, 3-A18NI enhanced V. cholerae colonization mimicking the phenotype of chemotaxis mutants that exhibit counterclockwise-biased flagellum rotation. PMID:28392408

  6. Array biosensor for detection of toxins

    Science.gov (United States)

    Ligler, Frances S.; Taitt, Chris Rowe; Shriver-Lake, Lisa C.; Sapsford, Kim E.; Shubin, Yura; Golden, Joel P.

    2003-01-01

    The array biosensor is capable of detecting multiple targets rapidly and simultaneously on the surface of a single waveguide. Sandwich and competitive fluoroimmunoassays have been developed to detect high and low molecular weight toxins, respectively, in complex samples. Recognition molecules (usually antibodies) were first immobilized in specific locations on the waveguide and the resultant patterned array was used to interrogate up to 12 different samples for the presence of multiple different analytes. Upon binding of a fluorescent analyte or fluorescent immunocomplex, the pattern of fluorescent spots was detected using a CCD camera. Automated image analysis was used to determine a mean fluorescence value for each assay spot and to subtract the local background signal. The location of the spot and its mean fluorescence value were used to determine the toxin identity and concentration. Toxins were measured in clinical fluids, environmental samples and foods, with minimal sample preparation. Results are shown for rapid analyses of staphylococcal enterotoxin B, ricin, cholera toxin, botulinum toxoids, trinitrotoluene, and the mycotoxin fumonisin. Toxins were detected at levels as low as 0.5 ng mL(-1).

  7. Pertussis toxin treatment of whole blood. A novel approach to assess G protein function in congestive heart failure

    NARCIS (Netherlands)

    Maisel, A. S.; Michel, M. C.; Insel, P. A.; Ennis, C.; Ziegler, M. G.; Phillips, C.

    1990-01-01

    This study was designed to assess G protein function in mononuclear leukocytes (MNL) of patients with congestive heart failure (CHF). MNL membranes were ADP-ribosylated in vitro in the presence of pertussis or cholera toxin. The amount of pertussis toxin substrates did not differ significantly

  8. Review of the inhibition of biological activities of food-related selected toxins by natural compounds.

    Science.gov (United States)

    Friedman, Mendel; Rasooly, Reuven

    2013-04-23

    There is a need to develop food-compatible conditions to alter the structures of fungal, bacterial, and plant toxins, thus transforming toxins to nontoxic molecules. The term 'chemical genetics' has been used to describe this approach. This overview attempts to survey and consolidate the widely scattered literature on the inhibition by natural compounds and plant extracts of the biological (toxicological) activity of the following food-related toxins: aflatoxin B1, fumonisins, and ochratoxin A produced by fungi; cholera toxin produced by Vibrio cholerae bacteria; Shiga toxins produced by E. coli bacteria; staphylococcal enterotoxins produced by Staphylococcus aureus bacteria; ricin produced by seeds of the castor plant Ricinus communis; and the glycoalkaloid α-chaconine synthesized in potato tubers and leaves. The reduction of biological activity has been achieved by one or more of the following approaches: inhibition of the release of the toxin into the environment, especially food; an alteration of the structural integrity of the toxin molecules; changes in the optimum microenvironment, especially pH, for toxin activity; and protection against adverse effects of the toxins in cells, animals, and humans (chemoprevention). The results show that food-compatible and safe compounds with anti-toxin properties can be used to reduce the toxic potential of these toxins. Practical applications and research needs are suggested that may further facilitate reducing the toxic burden of the diet. Researchers are challenged to (a) apply the available methods without adversely affecting the nutritional quality, safety, and sensory attributes of animal feed and human food and (b) educate food producers and processors and the public about available approaches to mitigating the undesirable effects of natural toxins that may present in the diet.

  9. 3-Amino 1,8-naphthalimide, a structural analog of the anti-cholera drug virstatin inhibits chemically-biased swimming and swarming motility in vibrios.

    Science.gov (United States)

    Wang, Hongxia; Silva, Anisia J; Benitez, Jorge A

    2017-06-01

    A screen for inhibitors of Vibrio cholerae motility identified the compound 3-amino 1,8-naphthalimide (3-A18NI), a structural analog of the cholera drug virstatin. Similar to virstatin, 3-A18NI diminished cholera toxin production. In contrast, 3-A18NI impeded swimming and/or swarming motility of V. cholerae and V. parahemolyticus suggesting that it could target the chemotaxis pathway shared by the polar and lateral flagellar system of vibrios. 3-A18NI did not inhibit the expression of V. cholerae major flagellin FlaA or the assembly of its polar flagellum. Finally, 3-A18NI enhanced V. cholerae colonization mimicking the phenotype of chemotaxis mutants that exhibit counterclockwise-biased flagellum rotation. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  10. Temperature and cholera toxin B are factors that influence formation of membrane nanotubes in RT4 and T24 urothelial cancer cell lines

    Directory of Open Access Journals (Sweden)

    Doron Kabaso

    2011-03-01

    Full Text Available Doron Kabaso1*, Maruša Lokar1*, Veronika Kralj-Iglic2, Peter Veranic3, Aleš Iglic11Laboratory of Biophysics, Faculty of Electrical Engineering, 2Laboratory of Clinical Biophysics, Faculty of Medicine, 3Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia; *These two authors equally share the first authorshipAbstract: The growth of membrane nanotubes is crucial for intercellular communication in both normal development and pathological conditions. Therefore, identifying factors that influence their stability and formation are important for both basic research and in development of potential treatments of pathological states. Here we investigate the effect of cholera toxin B (CTB and temperature on two pathological model systems: urothelial cell line RT4, as a model system of a benign tumor, and urothelial cell line T24, as a model system of a metastatic tumor. In particular, the number of intercellular membrane nanotubes (ICNs; ie, membrane nanotubes that bridge neighboring cells was counted. In comparison with RT4 cells, we reveal a significantly higher number in the density of ICNs in T24 cells not derived from RT4 without treatments (P = 0.005, after 20 minutes at room temperature (P = 0.0007, and following CTB treatment (P = 0.000025. The binding of CTB to GM1–lipid complexes in membrane exvaginations or tips of membrane nanotubes may reduce the positive spontaneous (intrinsic curvature of GM1–lipid complexes, which may lead to lipid mediated attractive interactions between CTB–GM1–lipid complexes, their aggregation and consequent formation of enlarged spherical tips of nanotubes. The binding of CTB to GM1 molecules in the outer membrane leaflet of membrane exvaginations and tips of membrane nanotubes may also increase the area difference between the two leaflets and in this way facilitate the growth of membrane nanotubes.Keywords: cancer cells, membrane nanotubes, cholera toxin

  11. Vibrio cholerae O1 secretes an extracellular matrix in response to antibody-mediated agglutination.

    Directory of Open Access Journals (Sweden)

    Danielle E Baranova

    Full Text Available Vibrio cholerae O1 is one of two serogroups responsible for epidemic cholera, a severe watery diarrhea that occurs after the bacterium colonizes the human small intestine and secretes a potent ADP-ribosylating toxin. Immunity to cholera is associated with intestinal anti-lipopolysaccharide (LPS antibodies, which are known to inhibit V. cholerae motility and promote bacterial cell-cell crosslinking and aggregation. Here we report that V. cholerae O1 classical and El Tor biotypes produce an extracellular matrix (ECM when forcibly immobilized and agglutinated by ZAC-3 IgG, an intestinally-derived monoclonal antibody (MAb against the core/lipid A region of LPS. ECM secretion, as demonstrated by crystal violet staining and scanning electron microscopy, occurred within 30 minutes of antibody exposure and peaked by 3 hours. Non-motile mutants of V. cholerae did not secrete ECM following ZAC-3 IgG exposure, even though they were susceptible to agglutination. The ECM was enriched in O-specific polysaccharide (OSP but not Vibrio polysaccharide (VPS. Finally, we demonstrate that ECM production by V. cholerae in response to ZAC-3 IgG was associated with bacterial resistant to a secondary complement-mediated attack. In summary, we propose that V. cholerae O1, upon encountering anti-LPS antibodies in the intestinal lumen, secretes an ECM (or O-antigen capsule possibly as a strategy to shield itself from additional host immune factors and to exit an otherwise inhospitable host environment.

  12. Critical Analysis of Compositions and Protective Efficacies of Oral Killed Cholera Vaccines

    Science.gov (United States)

    2014-01-01

    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

  13. Influence of tunicamycin, sialidase, and cholera toxin on gangliosides and T-lymphocyte responses to interleukin 2

    International Nuclear Information System (INIS)

    Semmes, O.J.; Bailey, J.M.; Merritt, W.D.

    1986-01-01

    The authors have shown that gangliosides inhibit interleukin 2 (IL 2)-dependent proliferation of murine T cells. Tunicamycin (TM), sialidase, and cholera toxin-β subunit (β-CT) are known modulators of cell surface glycoconjugates. To test the possible role of endogenous gangliosides in T cell responses to IL-2, the effect of these agents on ganglioside expression and cell proliferation was studied. Gangliosides were labelled for 24 hrs with 3 H-glucosamine/galactose in the presence of IL-2 and purified sialidase, TM or β-CT. Gangliosides were isolated and the species separated by TLC. Alternatively, proliferation was assayed by 3 H-thymidine uptake after 48 hrs culture. TM treatment at a concentration (10 μg/ml) that completely inhibited proliferation resulted in a 86% reduction of incorporation of saccharide precursors into gangliosides compared to a 50% reduction into proteins. Sialidase treatment (0.1 IU/ml) resulted in a 70% inhibition of proliferation and 30% reduction of radiolabel into gangliosides, of which 3 species were specifically reduced. β-CT, which binds to GM 1 and to a lesser extent GD/sub 1a/, caused a 50% reduction in proliferation response at 35 units/ml. The results support the hypothesis that gangliosides are involved in IL-2-dependent proliferation

  14. 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.

  15. From cholera to corals: Viruses as drivers of virulence in a major coral bacterial pathogen

    KAUST Repository

    Weynberg, Karen D.; Voolstra, Christian R.; Neave, Matthew J.; Buerger, Patrick; van Oppen, Madeleine J. H.

    2015-01-01

    Disease is an increasing threat to reef-building corals. One of the few identified pathogens of coral disease is the bacterium Vibrio coralliilyticus. In Vibrio cholerae, infection by a bacterial virus (bacteriophage) results in the conversion of non-pathogenic strains to pathogenic strains and this can lead to cholera pandemics. Pathogenicity islands encoded in the V. cholerae genome play an important role in pathogenesis. Here we analyse five whole genome sequences of V. coralliilyticus to examine whether virulence is similarly driven by horizontally acquired elements. We demonstrate that bacteriophage genomes encoding toxin genes with homology to those found in pathogenic V. cholerae are integrated in V. coralliilyticus genomes. Virulence factors located on chromosomal pathogenicity islands also exist in some strains of V. coralliilyticus. The presence of these genetic signatures indicates virulence in V. coralliilyticus is driven by prophages and other horizontally acquired elements. Screening for pathogens of coral disease should target conserved regions in these elements.

  16. Toxin-mediated effects on the innate mucosal defenses: implications for enteric vaccines

    DEFF Research Database (Denmark)

    Glenn, Gregory M; Francis, David H; Danielsen, E Michael

    2009-01-01

    mucosal barrier as a key step in enteric pathogen survival. We review key observations relevant to the roles of LT and cholera toxin in protective immunity and the effects of these toxins on innate mucosal defenses. We suggest either that toxin-mediated fluid secretion mechanically disrupts the mucus...... layer or that toxins interfere with innate mucosal defenses by other means. Such a breach gives pathogens access to the enterocyte, leading to binding and pathogenicity by enterotoxigenic E. coli (ETEC) and other organisms. Given the common exposure to LT(+) ETEC by humans visiting or residing...... unexpectedly broad protective effects against LT(+) ETEC and mixed infections when using a toxin-based enteric vaccine. If toxins truly exert barrier-disruptive effects as a key step in pathogenesis, then a return to classic toxin-based vaccine strategies for enteric disease is warranted and can be expected...

  17. Caracterización de aislamientos de Vibrio cholerae no-O1, no-O139 asociados a cuadros de diarrea Characterization of Vibrio cholerae non-O1 and non-O139 isolates associated with diarrhea

    Directory of Open Access Journals (Sweden)

    S. González Fraga

    2009-03-01

    its virulence factors by PCR, antimicrobial susceptibility patterns and genetic diversity by pulsed-field gel electrophoresis. Eight virulence patterns were obtained although no isolate was positive for the cholera toxin or the thermostable toxin. Four isolates were positive for the type three secretion system. The 17.6% of the isolates were resistant or intermediate to ampicillin and 5.9% were resistant to trimethoprim-sulfamethoxazole. By SfiI-PFGE, all isolates were genetically very diverse, as 27 different patterns were identified in 29 typeable isolates by pulsed-field gel electrophoresis. Although it has a low incidence, V. cholerae continues to be a causative agent of diarrhea in children, who are affected by a variety of circulating strains of V. cholerae non-O1, non-O139.

  18. Chloroplast-derived vaccine antigens confer dual immunity against cholera and malaria by oral or injectable delivery.

    Science.gov (United States)

    Davoodi-Semiromi, Abdoreza; Schreiber, Melissa; Nalapalli, Samson; Verma, Dheeraj; Singh, Nameirakpam D; Banks, Robert K; Chakrabarti, Debopam; Daniell, Henry

    2010-02-01

    Cholera and malaria are major diseases causing high mortality. The only licensed cholera vaccine is expensive; immunity is lost in children within 3 years and adults are not fully protected. No vaccine is yet available for malaria. Therefore, in this study, the cholera toxin-B subunit (CTB) of Vibrio cholerae fused to malarial vaccine antigens apical membrane antigen-1 (AMA1) and merozoite surface protein-1 (MSP1) was expressed in lettuce and tobacco chloroplasts. Southern blot analysis confirmed homoplasmy and stable integration of transgenes. CTB-AMA1 and CTB-MSP1 fusion proteins accumulated up to 13.17% and 10.11% (total soluble protein, TSP) in tobacco and up to 7.3% and 6.1% (TSP) in lettuce, respectively. Nine groups of mice (n = 10/group) were immunized subcutaneously (SQV) or orally (ORV) with purified antigens or transplastomic tobacco leaves. Significant levels of antigen-specific antibody titres of immunized mice completely inhibited proliferation of the malarial parasite and cross-reacted with the native parasite proteins in immunoblots and immunofluorescence studies. Protection against cholera toxin challenge in both ORV (100%) and SQV (89%) mice correlated with CTB-specific titres of intestinal, serum IgA and IgG1 in ORV and only IgG1 in SQV mice, but no other immunoglobulin. Increasing numbers of interleukin-10(+) T cell but not Foxp3(+) regulatory T cells, suppression of interferon-gamma and absence of interleukin-17 were observed in protected mice, suggesting that immunity is conferred via the Tr1/Th2 immune response. Dual immunity against two major infectious diseases provided by chloroplast-derived vaccine antigens for long-term (>300 days, 50% of mouse life span) offers a realistic platform for low cost vaccines and insight into mucosal and systemic immunity.

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

    Directory of Open Access Journals (Sweden)

    Soroor Erfanimanesh

    2014-10-01

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

  20. Activation of AMPK inhibits cholera toxin stimulated chloride secretion in human and murine intestine.

    Directory of Open Access Journals (Sweden)

    Ailín C Rogers

    Full Text Available Increased intestinal chloride secretion through chloride channels, such as the cystic fibrosis transmembrane conductance regulator (CFTR, is one of the major molecular mechanisms underlying enterotoxigenic diarrhea. It has been demonstrated in the past that the intracellular energy sensing kinase, the AMP-activated protein kinase (AMPK, can inhibit CFTR opening. We hypothesized that pharmacological activation of AMPK can abrogate the increased chloride flux through CFTR occurring during cholera toxin (CTX mediated diarrhea. Chloride efflux was measured in isolated rat colonic crypts using real-time fluorescence imaging. AICAR and metformin were used to activate AMPK in the presence of the secretagogues CTX or forskolin (FSK. In order to substantiate our findings on the whole tissue level, short-circuit current (SCC was monitored in human and murine colonic mucosa using Ussing chambers. Furthermore, fluid accumulation was measured in excised intestinal loops. CTX and forskolin (FSK significantly increased chloride efflux in isolated colonic crypts. The increase in chloride efflux could be offset by using the AMPK activators AICAR and metformin. In human and mouse mucosal sheets, CTX and FSK increased SCC. AICAR and metformin inhibited the secretagogue induced rise in SCC, thereby confirming the findings made in isolated crypts. Moreover, AICAR decreased CTX stimulated fluid accumulation in excised intestinal segments. The present study suggests that pharmacological activation of AMPK effectively reduces CTX mediated increases in intestinal chloride secretion, which is a key factor for intestinal water accumulation. AMPK activators may therefore represent a supplemental treatment strategy for acute diarrheal illness.

  1. Modulating the Global Response Regulator, LuxO of V. cholerae Quorum Sensing System Using a Pyrazine Dicarboxylic Acid Derivative (PDCApy: An Antivirulence Approach

    Directory of Open Access Journals (Sweden)

    M. Hema

    2017-10-01

    Full Text Available Vibrio cholerae is a Gram-negative pathogen which causes acute diarrhoeal disease, cholera by the expression of virulence genes through quorum sensing (QS mechanism. The QS circuit of V. cholerae is controlled by the global quorum regulator, LuxO, which at low cell density (LCD state produces major virulence factors such as, toxin co-regulated pilus (TCP and cholera toxin (CT to mediate infection. On the contrary, at the high cell density (HCD state the virulent genes are downregulated and the vibrios are detached from the host intestinal epithelial cells, promoted by HapA protease. Hence, targeting the global regulator LuxO would be a promising approach to modulate the QS to curtail V. cholerae pathogenesis. In our earlier studies, LuxO targeted ligand, 2,3 pyrazine dicarboxylic acid (PDCA and its derivatives having desired pharmacophore properties were chemically synthesized and were shown to have biofilm inhibition as well as synergistic activity with the conventionally used antibiotics. In the present study, the QS modulatory effect of the PDCA derivative with pyrrolidine moiety designated as PDCApy against the V. cholerae virulence gene expression was analyzed at various growth phases. The data significantly showed a several fold reduction in the expression of the genes, tcp and ct whereas the expression of hapR was upregulated at the LCD state. In addition, PDCApy reduced the adhesion and invasion of the vibrios onto the INT407 intestinal cell lines. Collectively, our data suggest that PDCApy could be a potential QS modulator (QSM for the antivirulence therapeutic approach.

  2. Cholera Treatment

    Science.gov (United States)

    ... Diagnosis and Detection Laboratory Testing for Cholera Treatment Rehydration Therapy Antibiotic Treatment Zinc Treatment Prevention & Control Five ... page for current cholera treatment recommendations. Cholera Treatments Rehydration therapy , meaning prompt restoration of lost fluids and ...

  3. The aquatic environment as a reservoir of Vibrio cholerae O1 in hydrographic basins of the state of Pernambuco, Brazil.

    Science.gov (United States)

    Mendes-Marques, Carina Lucena; Silveira Filho, Vladimir da Mota; da Costa, Ana Paula Rocha; Nunes, Mariana de Lira; da Silva Filho, Sandoval Vieira; Figueirôa, Ângela Cristina Torres de Araújo; Hofer, Ernesto; de Almeida, Alzira Maria Paiva; Leal, Nilma Cintra

    2013-01-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Carina Lucena Mendes-Marques

    2013-01-01

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

  5. Update: cholera--Western Hemisphere, and recommendations for treatment of cholera.

    Science.gov (United States)

    1991-08-16

    Epidemic cholera appeared in Peru in January 1991 and subsequently spread to Ecuador, Colombia, Chile, Brazil, Mexico, and Guatemala. Cholera can be a severe, life-threatening illness but is highly preventable and easily treated; however, few health-care practitioners in the United States have experience identifying and treating cholera. This report provides an update on cholera in the Western Hemisphere and provides recommendations on the clinical diagnosis and treatment of cholera in the United States.

  6. A Role for the Mannose-Sensitive Hemagglutinin in Biofilm Formation by Vibrio cholerae El Tor

    Science.gov (United States)

    Watnick, Paula I.; Fullner, Karla Jean; Kolter, Roberto

    1999-01-01

    While much has been learned regarding the genetic basis of host-pathogen interactions, less is known about the molecular basis of a pathogen’s survival in the environment. Biofilm formation on abiotic surfaces represents a survival strategy utilized by many microbes. Here it is shown that Vibrio cholerae El Tor does not use the virulence-associated toxin-coregulated pilus to form biofilms on borosilicate but rather uses the mannose-sensitive hemagglutinin (MSHA) pilus, which plays no role in pathogenicity. In contrast, attachment of V. cholerae to chitin is shown to be independent of the MSHA pilus, suggesting divergent pathways for biofilm formation on nutritive and nonnutritive abiotic surfaces. PMID:10348878

  7. Effects of cholera toxin on the potential difference and motor responses induced by distension in the rat proximal small intestine in vivo.

    Science.gov (United States)

    Kordasti, Shirin; Sapnara, Maria; Thomas, Evan A; Lindstrom, Erik; Forsman, Mikael; Bornstein, Joel C; Sjövall, Henrik

    2006-05-01

    Cholera toxin (CT) may induce uncontrolled firing in recurrent networks of secretomotor neurons in the submucous plexus. This hypothesis was tested in chloralose-anesthetized rats in vivo. The secretory reflex response to graded intestinal distension was measured with or without prior exposure to luminal CT. The transmural potential difference (PD) was used as a marker for electrogenic chloride secretion. In controls, distension increased PD, and this response was reduced by the neural blocker tetrodotoxin given serosally and the vasoactive intestinal peptide (VIP) receptor antagonist [4Cl-d-Phe(6),Leu(17)]VIP (2 mug.min(-1).kg(-1) iv) but unaffected by the serotonin 5-HT(3) receptor antagonist granisetron, by the nicotinic receptor antagonist hexamethonium, by the muscarinic receptor antagonist atropine, or by the cyclooxygenase inhibitor indomethacin. Basal PD increased significantly with time in CT-exposed segments, an effect blocked by granisetron, by indomethacin, and by [4Cl-d-Phe(6),Leu(17)]VIP but not by hexamethonium or atropine. In contrast, once the increased basal PD produced by CT was established, [4Cl-d-Phe(6),Leu(17)]VIP and indomethacin had no significant effect, whereas granisetron and hexamethonium markedly depressed basal PD. CT significantly reduced the increase in PD produced by distension, an effect reversed by granisetron, indomethacin, and atropine. CT also activated a specific motility response to distension, repeated cluster contractions, but only in animals pretreated with granisetron, indomethacin, or atropine. These data are compatible with the hypothesis that CT induces uncontrolled activity in submucous secretory networks. Development of this state depends on 5-HT(3) receptors, VIP receptors, and prostaglandin synthesis, whereas its maintenance depends on 5-HT(3) and nicotinic receptors but not VIP receptors. The motility effects of CT (probably reflecting myenteric activity) are partially suppressed via a mechanism involving 5-HT(3

  8. Novel type of specialized transduction for CTX phi or its satellite phage RS1 mediated by filamentous phage VGJ phi in Vibrio cholerae.

    Science.gov (United States)

    Campos, Javier; Martínez, Eriel; Marrero, Karen; Silva, Yussuan; Rodríguez, Boris L; Suzarte, Edith; Ledón, Talena; Fando, Rafael

    2003-12-01

    The main virulence factor of Vibrio cholerae, the cholera toxin, is encoded by the ctxAB operon, which is contained in the genome of the lysogenic filamentous phage CTX phi. This phage transmits ctxAB genes between V. cholerae bacterial populations that express toxin-coregulated pilus (TCP), the CTX phi receptor. In investigating new forms of ctxAB transmission, we found that V. cholerae filamentous phage VGJ phi, which uses the mannose-sensitive hemagglutinin (MSHA) pilus as a receptor, transmits CTX phi or its satellite phage RS1 by an efficient and highly specific TCP-independent mechanism. This is a novel type of specialized transduction consisting in the site-specific cointegration of VGJ phi and CTX phi (or RS1) replicative forms to produce a single hybrid molecule, which generates a single-stranded DNA hybrid genome that is packaged into hybrid viral particles designated HybP phi (for the VGJ phi/CTX phi hybrid) and HybRS phi (for the VGJ phi/RS1 hybrid). The hybrid phages replicate by using the VGJ phi replicating functions and use the VGJ phi capsid, retaining the ability to infect via MSHA. The hybrid phages infect most tested strains more efficiently than CTX phi, even under in vitro optimal conditions for TCP expression. Infection and lysogenization with HybP phi revert the V. cholerae live attenuated vaccine strain 1333 to virulence. Our results reinforce that TCP is not indispensable for the acquisition of CTX phi. Thus, we discuss an alternative to the current accepted evolutionary model for the emergence of new toxigenic strains of V. cholerae and the importance of our findings for the development of an environmentally safer live attenuated cholera vaccine.

  9. [Mexican phenotype and genotype Vibrio cholerae 01].

    Science.gov (United States)

    Giono, S; Gutiérrez Cogno, L; Rodríguez Angeles, G; del Rio Zolezzi, A; Valdespino González, J L; Sepúlveda Amor, J

    1995-01-01

    This paper presents the phenotypical and genotypical characterization of 26922 Vibrio cholerae 01 strains isolated in Mexico from 1991 to 1993. All strains isolated were El Tor biovar. Strains were sensitive to antibiotics excluding furazolidone, streptomycin and sulfisoxasole to which we found resistance in 97% and we are using this characteristic as epidemiological markers. We detected a marked change in frequency of Inaba serotype from 1991, when it was dominant, with 99.5%, until 1992 when Ogawa serotype turned to be dominant with 95% of isolates. All Vibrio cholerae 01 strains, except one Ogawa strain, were to igenic, and V. choleraeno 01 were not toxigenic by ELISA, PCR and cell culture tests. Dominant ribotype was 5, but we found some strains with 6a pattern and two with ribotype 12. We are searching for ribotype 2 among hemolytic strains in order to learn if there is any relation to Gulf Coast strains prevalent in the USA, but until now we have not found any V. cholerae ribotype 2 in our isolates. Even if rapid tests are recommended for immediate diagnosis of cholera, it is necessary to continue bacterial isolation in order to have strains for phenotyping and genotyping studies that may support epidemiological analysis.

  10. 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; Son, Mike S

    2016-01-01

    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 cholera

  11. A bistable switch and anatomical site control Vibrio cholerae virulence gene expression in the intestine.

    Directory of Open Access Journals (Sweden)

    Alex T Nielsen

    2010-09-01

    Full Text Available A fundamental, but unanswered question in host-pathogen interactions is the timing, localization and population distribution of virulence gene expression during infection. Here, microarray and in situ single cell expression methods were used to study Vibrio cholerae growth and virulence gene expression during infection of the rabbit ligated ileal loop model of cholera. Genes encoding the toxin-coregulated pilus (TCP and cholera toxin (CT were powerfully expressed early in the infectious process in bacteria adjacent to epithelial surfaces. Increased growth was found to co-localize with virulence gene expression. Significant heterogeneity in the expression of tcpA, the repeating subunit of TCP, was observed late in the infectious process. The expression of tcpA, studied in single cells in a homogeneous medium, demonstrated unimodal induction of tcpA after addition of bicarbonate, a chemical inducer of virulence gene expression. Striking bifurcation of the population occurred during entry into stationary phase: one subpopulation continued to express tcpA, whereas the expression declined in the other subpopulation. ctxA, encoding the A subunit of CT, and toxT, encoding the proximal master regulator of virulence gene expression also exhibited the bifurcation phenotype. The bifurcation phenotype was found to be reversible, epigenetic and to persist after removal of bicarbonate, features consistent with bistable switches. The bistable switch requires the positive-feedback circuit controlling ToxT expression and formation of the CRP-cAMP complex during entry into stationary phase. Key features of this bistable switch also were demonstrated in vivo, where striking heterogeneity in tcpA expression was observed in luminal fluid in later stages of the infection. When this fluid was diluted into artificial seawater, bacterial aggregates continued to express tcpA for prolonged periods of time. The bistable control of virulence gene expression points to a

  12. Traffic of antibody-secreting cells after immunization with a liposome-associated, CpG-ODN-adjuvanted oral cholera vaccine.

    Science.gov (United States)

    Somroop, Srinuan; Tongtawe, Pongsri; Chaisri, Urai; Tapchaisri, Pramuan; Chongsa-nguan, Manas; Srimanote, Potjanee; Chaicumpa, Wanpen

    2006-12-01

    An oral cholera vaccine made up of heat-treated recombinant cholera toxin (rCT), V. cholerae lipopolysaccharide (LPS), and recombinant toxin-co-regulated pili subunit A (rTcpA), entrapped in liposomes in the presence of unmethylated bacterial CpG-DNA (ODN#1826) was used to orally immunize a group of eight week old rats. A booster dose was given 14 days later. Control rats received placebo (vaccine diluent). The kinetics of the immune response were investigated by enumerating the antigen specific-antibody secreting cells (ASC) in the blood circulation and intestinal lamina propria using the ELISPOT assay and a histo-immunofluorescence assay (IFA), respectively. ASC of all antigenic specificities were detected in the blood of the vaccinated rats as early as two days after the booster dose. The numbers of LPS-ASC and TcpA-ASC in the blood were at their peak at day 3 post booster while the number of CT-ASC was highest at day 4 after the booster immunization. At day 13 post immunization, no ASC were detected in the blood. A several fold increase in the number of ASC of all antigenic specificities in the lamina propria above the background numbers of the control animals were found in all vaccinated rats at days 6 and 13 post booster (earlier and later time points were not studied). Vibriocidal antibody and specific antibodies to CT, LPS and TcpA were detected in 57.1% and 52.4%, 14.3%, and 19.0% of the orally vaccinated rats, respectively. The data indicated that rats orally primed with the vaccine could produce a rapid anamnestic response after re-exposure to the V. cholerae antigens. Thus, a single dose of the vaccine is expected to elicit a similar anamnestic immune response in people from cholera endemic areas who have been naturally primed to V. cholerae antigens, while two doses at a 14 day interval should be adequate for a traveler to a disease endemicarea.

  13. Low doses of cholera toxin and its mediator cAMP induce CTLA-2 secretion by dendritic cells to enhance regulatory T cell conversion.

    Directory of Open Access Journals (Sweden)

    Cinthia Silva-Vilches

    Full Text Available Immature or semi-mature dendritic cells (DCs represent tolerogenic maturation stages that can convert naive T cells into Foxp3+ induced regulatory T cells (iTreg. Here we found that murine bone marrow-derived DCs (BM-DCs treated with cholera toxin (CT matured by up-regulating MHC-II and costimulatory molecules using either high or low doses of CT (CThi, CTlo or with cAMP, a known mediator CT signals. However, all three conditions also induced mRNA of both isoforms of the tolerogenic molecule cytotoxic T lymphocyte antigen 2 (CTLA-2α and CTLA-2β. Only DCs matured under CThi conditions secreted IL-1β, IL-6 and IL-23 leading to the instruction of Th17 cell polarization. In contrast, CTlo- or cAMP-DCs resembled semi-mature DCs and enhanced TGF-β-dependent Foxp3+ iTreg conversion. iTreg conversion could be reduced using siRNA blocking of CTLA-2 and reversely, addition of recombinant CTLA-2α increased iTreg conversion in vitro. Injection of CTlo- or cAMP-DCs exerted MOG peptide-specific protective effects in experimental autoimmune encephalomyelitis (EAE by inducing Foxp3+ Tregs and reducing Th17 responses. Together, we identified CTLA-2 production by DCs as a novel tolerogenic mediator of TGF-β-mediated iTreg induction in vitro and in vivo. The CT-induced and cAMP-mediated up-regulation of CTLA-2 also may point to a novel immune evasion mechanism of Vibrio cholerae.

  14. Immunogenicity and protective efficacy of rotavirus VP8* fused to cholera toxin B subunit in a mouse model.

    Science.gov (United States)

    Xue, Miaoge; Yu, Linqi; Jia, Lianzhi; Li, Yijian; Zeng, Yuanjun; Li, Tingdong; Ge, Shengxiang; Xia, Ningshao

    2016-11-01

    In attempts to develop recombinant subunit vaccines against rotavirus disease, it was previously shown that the N-terminal truncated VP8* protein, VP8-1 (aa26-231), is a good vaccine candidate when used for immunization in combination with Freund's adjuvant. However, this protein stimulated only weak immune response when aluminum hydroxide was used as an adjuvant. In this study, the nontoxic B subunit of cholera toxin (CTB) was employed as intra-molecular adjuvant to improve the immunogenicity of VP8-1. Both, the N-terminal and C-terminal fusion proteins, were purified to homogeneity, at which stage they formed pentamers, and showed significantly higher immunogenicity and protective efficacy than a VP8-1/aluminum hydroxide mixture in a mouse model. Compared to VP8-1-CTB, CTB-VP8-1 showed higher binding activity to both, GM1 and the conformation sensitive neutralizing monoclonal antibodies specific to VP8. More importantly, CTB-VP8-1 elicited higher titers of neutralizing antibodies and conferred higher protective efficacy than VP8-1-CTB. Therefore, the protein CTB-VP8-1, with enhanced immunogenicity and immunoprotectivity, could be considered as a viable candidate for further development of an alternative, replication-incompetent, parenterally administered vaccine against rotavirus disease.

  15. Glucose- but not rice-based oral rehydration therapy enhances the production of virulence determinants in the human pathogen Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    Juliane Kühn

    2014-12-01

    Full Text Available Despite major attempts to prevent cholera transmission, millions of people worldwide still must address this devastating disease. Cholera research has so far mainly focused on the causative agent, the bacterium Vibrio cholerae, or on disease treatment, but rarely were results from both fields interconnected. Indeed, the treatment of this severe diarrheal disease is mostly accomplished by oral rehydration therapy (ORT, whereby water and electrolytes are replenished. Commonly distributed oral rehydration salts also contain glucose. Here, we analyzed the effects of glucose and alternative carbon sources on the production of virulence determinants in the causative agent of cholera, the bacterium Vibrio cholerae during in vitro experimentation. We demonstrate that virulence gene expression and the production of cholera toxin are enhanced in the presence of glucose or similarly transported sugars in a ToxR-, TcpP- and ToxT-dependent manner. The virulence genes were significantly less expressed if alternative non-PTS carbon sources, including rice-based starch, were utilized. Notably, even though glucose-based ORT is commonly used, field studies indicated that rice-based ORT performs better. We therefore used a spatially explicit epidemiological model to demonstrate that the better performing rice-based ORT could have a significant impact on epidemic progression based on the recent outbreak of cholera in Haiti. Our results strongly support a change of carbon source for the treatment of cholera, especially in epidemic settings.

  16. Low-dose oral tolerance due to antigen in the diet suppresses differentially the cholera toxin-adjuvantized IgE, IgA and IgG response

    DEFF Research Database (Denmark)

    Christensen, Hanne Risager; Kjær, Tanja; Frøkiær, Hanne

    2003-01-01

    Background: Cholera toxin (CT) is used as a mucosal adjuvant amongst other applications for studying food allergy because oral administration of antigen with CT induces an antigen-specific type 2 response, including IgE and IgA production. Priorly established oral tolerance due to antigen...... soy-trypsin inhibitor (KSTI) (F0 mice) and mice fed a soy-free diet (F2 mice) were orally immunized with KSTI and CT. KSTI-specific serum IgG1, IgG2a, IgA and IgE and fecal IgA were monitored. KSTI-stimulated cell proliferation and interleukin (IL)-6 production were determined. Results: The anti...... immunizations. However, cell proliferation and IL-6 production were clearly suppressed even after five immunizations. Conclusions: Priorly established low-dose oral tolerance considerably suppressed the CT-adjuvantized KSTI-specific IgE, IgA and cellular immune response but only weakly and transiently the Ig...

  17. Mutations in the Histone-like Nucleoid Structuring Regulatory Gene (hns) Decrease the Adherence of Shiga Toxin-producing Escherichia coli 091:H21 Strain B2F1 to Human Colonic Epithelial Cells and Increase the Production of Hemolysin

    Science.gov (United States)

    1999-10-19

    osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J. Bacteriol. 170:2575-2583. Mobley, H. L., D. M. Green...produced by ETEC organisms is homologous to the toxin encoded by Y: cholerae . These toxins are the primary cause of the watery diarrhea associated with ETEC...Escherichia coli as a cause ofdiarrhea among children in Mexico . J. Clin. Microbiol. 25:1913-1919. Maurelli, A. T., and P. J. Sansonetti. 1988

  18. Antisecretory activity from the flowers of Chiranthodendron pentadactylon and its flavonoids on intestinal fluid accumulation induced by Vibrio cholerae toxin in rats.

    Science.gov (United States)

    Velázquez, Claudia; Calzada, Fernando; Esquivel, Baldomero; Barbosa, Elizabeth; Calzada, Samuel

    2009-12-10

    The flowers of Chiranthodendron pentadactylon Larreat. (Sterculiaceae) has been traditionally used as folk medicine in Mexico for the treatment of gastrointestinal disorders such as diarrhea and dysentery. This study aimed to assess the antisecretory activity which supports the therapeutic use of Chiranthodendron pentadactylon and its flavonoids to treat diarrhea. The methanol extract of Chiranthodendron pentadactylon, subsequent fractions, and flavonoids were evaluated on cholera toxin-induced intestinal secretion in rat jejunal loops model. Three antisecretory flavonoids were isolated by bioassay-guided purification, namely, isoquercitrin 3, (+)-catechin 4 and (-)-epicatechin 5. Among them, epicatechin exhibited the most potent antisecretory activity with ID(50) of 8.3 microM/kg. Its potency was close that of to loperamide (ID(50) 6.1 microM/kg), drug used as control. Isoquercitrin (ID(50) 19.2 microM/kg) and catechin (ID(50) 51.7 microM/kg) showed moderate and weak activity, respectively. The results of the present study lend some support to the anecdotal report for the traditional use of the flowers of Chiranthodendron pentadactylon in the control of dysentery.

  19. Characterization of Vibrio cholerae O1 El Tor Biotype Variant Clinical Isolates from Bangladesh and Haiti, Including a Molecular Genetic Analysis of Virulence Genes ▿

    Science.gov (United States)

    Son, Mike S.; Megli, Christina J.; Kovacikova, Gabriela; Qadri, Firdausi; Taylor, Ronald K.

    2011-01-01

    Vibrio cholerae serogroup O1, the causative agent of the diarrheal disease cholera, is divided into two biotypes: classical and El Tor. Both biotypes produce the major virulence factors toxin-coregulated pilus (TCP) and cholera toxin (CT). Although possessing genotypic and phenotypic differences, El Tor biotype strains displaying classical biotype traits have been reported and subsequently were dubbed El Tor variants. Of particular interest are reports of El Tor variants that produce various levels of CT, including levels typical of classical biotype strains. Here, we report the characterization of 10 clinical isolates from the International Centre for Diarrhoeal Disease Research, Bangladesh, and a representative strain from the 2010 Haiti cholera outbreak. We observed that all 11 strains produced increased CT (2- to 10-fold) compared to that of wild-type El Tor strains under in vitro inducing conditions, but they possessed various TcpA and ToxT expression profiles. Particularly, El Tor variant MQ1795, which produced the highest level of CT and very high levels of TcpA and ToxT, demonstrated hypervirulence compared to the virulence of El Tor wild-type strains in the infant mouse cholera model. Additional genotypic and phenotypic tests were conducted to characterize the variants, including an assessment of biotype-distinguishing characteristics. Notably, the sequencing of ctxB in some El Tor variants revealed two copies of classical ctxB, one per chromosome, contrary to previous reports that located ctxAB only on the large chromosome of El Tor biotype strains. PMID:21880975

  20. 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

  1. Cholera in Zimbabwe

    NARCIS (Netherlands)

    Pruyt, E.

    2009-01-01

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

  2. The global burden of cholera

    Science.gov (United States)

    Lopez, Anna Lena; You, Young Ae; Kim, Young Eun; Sah, Binod; Maskery, Brian; Clemens, John

    2012-01-01

    Abstract Objective To estimate the global burden of cholera using population-based incidence data and reports. Methods Countries with a recent history of cholera were classified as endemic or non-endemic, depending on whether they had reported cholera cases in at least three of the five most recent years. The percentages of the population in each country that lacked access to improved sanitation were used to compute the populations at risk for cholera, and incidence rates from published studies were applied to groups of countries to estimate the annual number of cholera cases in endemic countries. The estimates of cholera cases in non-endemic countries were based on the average numbers of cases reported from 2000 to 2008. Literature-based estimates of cholera case-fatality rates (CFRs) were used to compute the variance-weighted average cholera CFRs for estimating the number of cholera deaths. Findings About 1.4 billion people are at risk for cholera in endemic countries. An estimated 2.8 million cholera cases occur annually in such countries (uncertainty range: 1.4–4.3) and an estimated 87 000 cholera cases occur in non-endemic countries. The incidence is estimated to be greatest in children less than 5 years of age. Every year about 91 000 people (uncertainty range: 28 000 to 142 000) die of cholera in endemic countries and 2500 people die of the disease in non-endemic countries. Conclusion The global burden of cholera, as determined through a systematic review with clearly stated assumptions, is high. The findings of this study provide a contemporary basis for planning public health interventions to control cholera. PMID:22461716

  3. Household and Individual Risk Factors for Cholera among Cholera Vaccine Recipients in Rural Haiti.

    Science.gov (United States)

    Matias, Wilfredo R; Teng, Jessica E; Hilaire, Isabelle J; Harris, Jason B; Franke, Molly F; Ivers, Louise C

    2017-08-01

    Oral cholera vaccination was used as part of cholera control in Haiti, but the vaccine does not provide complete protection. We conducted secondary data analyses of a vaccine effectiveness study in Haiti to evaluate risk factors for cholera among cholera vaccine recipients. Individuals vaccinated against cholera that presented with acute watery diarrhea and had a stool sample positive for Vibrio cholerae O1 were included as cases. Up to four vaccinated individuals who did not present for treatment of diarrhea were included as controls for each case, and matched by location of residence, enrollment time, and age. We evaluated sociodemographic characteristics and risk factors for cholera. Univariable and multivariable logistic regression were performed to identify risk factors for cholera among vaccinees. Thirty-three vaccine recipients with culture-confirmed cholera were included as cases. One-hundred-and-seventeen of their matched controls reported receiving vaccine and were included as controls. In a multivariable analysis, self-reporting use of branded household water disinfection products as a means of treating water (adjusted relative risk [aRR] = 44.3, 95% confidence interval [CI] = 4.19-468.05, P = 0.002), and reporting having a latrine as the main household toilet (aRR = 4.22, 95% CI = 1.23-14.43, P = 0.02), were independent risk factors for cholera. Self-reporting always treating water (aRR = 0.09, 95% CI = 0.01-0.57, P = 0.01) was associated with protection against cholera. The field effectiveness of water, sanitation, and hygiene interventions used in combination with cholera vaccination in cholera control should be measured and monitored over time to identify and remediate shortcomings, and ensure successful impact on disease control.

  4. [Cholera in pediatrics].

    Science.gov (United States)

    Lezama-Basulto, L A; Mota-Hernández, F

    1993-09-01

    Cholerae is a grave and acute bacterial intestine infection which is caused by a bacilo, V. cholerae 01, that produces toxic products. Its clinical symptoms range from abundant liquid diarrhoea combined with vomiting and rapid dehydration. It is highly lethal when right treatment is not applied. There are also cases of cholera where victims do not show any symptoms of it, that is asymptomatic carriers. Any clinical suspicion of cholerae has to be corroborated by epidemiological data and its diagnostic confirmation should be done by isolating the bacteria, V. cholerae. When beginning the treatment, it is not necessary to confirm the diagnostic and this is based on the restitution of the liquids lost through vomiting and facing using any methods that are recommended for any other type of diarrhoea. The antimicrobial treatment is used only for grave cases. This present revision includes recent knowledge about cholerae emphasising on the effective management of cases through an adequate use of right treatment methods and also using the principal prevention measures against dissemination of this disease.

  5. Cost-effectiveness of oral cholera vaccine in a stable refugee population at risk for epidemic cholera and in a population with endemic cholera.

    OpenAIRE

    Murray, J.; McFarland, D. A.; Waldman, R. J.

    1998-01-01

    Recent large epidemics of cholera with high incidence and associated mortality among refugees have raised the question of whether oral cholera vaccines should be considered as an additional preventive measure in high-risk populations. The potential impact of oral cholera vaccines on populations prone to seasonal endemic cholera has also been questioned. This article reviews the potential cost-effectiveness of B-subunit, killed whole-cell (BS-WC) oral cholera vaccine in a stable refugee popula...

  6. Identifying cholera "hotspots" in Uganda: An analysis of cholera surveillance data from 2011 to 2016.

    Science.gov (United States)

    Bwire, Godfrey; Ali, Mohammad; Sack, David A; Nakinsige, Anne; Naigaga, Martha; Debes, Amanda K; Ngwa, Moise C; Brooks, W Abdullah; Garimoi Orach, Christopher

    2017-12-01

    Despite advance in science and technology for prevention, detection and treatment of cholera, this infectious disease remains a major public health problem in many countries in sub-Saharan Africa, Uganda inclusive. The aim of this study was to identify cholera hotspots in Uganda to guide the development of a roadmap for prevention, control and elimination of cholera in the country. We obtained district level confirmed cholera outbreak data from 2011 to 2016 from the Ministry of Health, Uganda. Population and rainfall data were obtained from the Uganda Bureau of Statistics, and water, sanitation and hygiene data from the Ministry of Water and Environment. A spatial scan test was performed to identify the significantly high risk clusters. Cholera hotspots were defined as districts whose center fell within a significantly high risk cluster or where a significantly high risk cluster was completely superimposed onto a district. A zero-inflated negative binomial regression model was employed to identify the district level risk factors for cholera. In total 11,030 cases of cholera were reported during the 6-year period. 37(33%) of 112 districts reported cholera outbreaks in one of the six years, and 20 (18%) districts experienced cholera at least twice in those years. We identified 22 districts as high risk for cholera, of which 13 were near a border of Democratic Republic of Congo (DRC), while 9 districts were near a border of Kenya. The relative risk of having cholera inside the high-risk districts (hotspots) were 2 to 22 times higher than elsewhere in the country. In total, 7 million people were within cholera hotspots. The negative binomial component of the ZINB model shows people living near a lake or the Nile river were at increased risk for cholera (incidence rate ratio, IRR = 0.98, 95% CI: 0.97 to 0.99, p cholera in a district (IRR = 0.99, 95% CI: 0.98 to 1.00, p = .02 and IRR = 1.02, 95% CI: 1.01 to 1.03, p cholera in the district. The study identified cholera

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

    OpenAIRE

    Choi, Seon Young; Rashed, Shah M.; Hasan, Nur A.; Alam, Munirul; Islam, Tarequl; Sadique, Abdus; Johura, Fatema-Tuz; Eppinger, Mark; Ravel, Jacques; Huq, Anwar; Cravioto, Alejandro; Colwell, Rita R.

    2016-01-01

    ABSTRACT 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...

  8. Tumor endothelium marker-8 based decoys exhibit superiority over capillary morphogenesis protein-2 based decoys as anthrax toxin inhibitors.

    Directory of Open Access Journals (Sweden)

    Chenguang Cai

    Full Text Available Anthrax toxin is the major virulence factor produced by Bacillus anthracis. The toxin consists of three protein subunits: protective antigen (PA, lethal factor, and edema factor. Inhibition of PA binding to its receptors, tumor endothelium marker-8 (TEM8 and capillary morphogenesis protein-2 (CMG2 can effectively block anthrax intoxication, which is particularly valuable when the toxin has already been overproduced at the late stage of anthrax infection, thus rendering antibiotics ineffectual. Receptor-like agonists, such as the mammalian cell-expressed von Willebrand factor type A (vWA domain of CMG2 (sCMG2, have demonstrated potency against the anthrax toxin. However, the soluble vWA domain of TEM8 (sTEM8 was ruled out as an anthrax toxin inhibitor candidate due to its inferior affinity to PA. In the present study, we report that L56A, a PA-binding-affinity-elevated mutant of sTEM8, could inhibit anthrax intoxication as effectively as sCMG2 in Fisher 344 rats. Additionally, pharmacokinetics showed that L56A and sTEM8 exhibit advantages over sCMG2 with better lung-targeting and longer plasma retention time, which may contribute to their enhanced protective ability in vivo. Our results suggest that receptor decoys based on TEM8 are promising anthrax toxin inhibitors and, together with the pharmacokinetic studies in this report, may contribute to the development of novel anthrax drugs.

  9. Molecular cloning, characterization, and expression of human ADP-ribosylation factors: Two guanine nucleotide-dependent activators of cholera toxin

    International Nuclear Information System (INIS)

    Bobak, D.A.; Nightingale, M.S.; Murtagh, J.J.; Price, S.R.; Moss, J.; Vaughan, M.

    1989-01-01

    ADP-ribosylation factors (ARFs) are small guanine nucleotide-binding proteins that enhance the enzymatic activities of cholera toxin. Two ARF cDNAs, ARF1 and ARF3, were cloned from a human cerebellum library. Based on deduced amino acid sequences and patterns of hybridization of cDNA and oligonucleotide probes with mammalian brain poly(A) + RNA, human ARF1 is the homologue of bovine ARF1. Human ARF3, which differs from bovine ARF1 and bovine ARF2, appears to represent a newly identified third type of ARF. Hybridization patterns of human ARF cDNA and clone-specific oligonucleotides with poly(A) + RNA are consistent with the presence of at least two, and perhaps four, separate ARF messages in human brain. In vitro translation of ARF1, ARF2, and ARF3 produced proteins that behaved, by SDS/PAGE, similar to a purified soluble brain ARF. Deduced amino acid sequences of human ARF1 and ARF3 contain regions, similar to those in other G proteins, that are believed to be involved in GTP binding and hydrolysis. ARFS also exhibit a modest degree of homology with a bovine phospholipase C. The observations reported here support the conclusion that the ARFs are members of a multigene family of small guanine nucleotide-binding proteins. Definition of the regulation of ARF mRNAs and of function(s) of recombinant ARF proteins will aid in the elucidation of the physiologic role(s) of ARFs

  10. Improving immunization approaches to cholera.

    Science.gov (United States)

    Saha, Amit; Rosewell, Alexander; Hayen, Andrew; MacIntyre, C Raina; Qadri, Firdausi

    2017-03-01

    Cholera's impact is greatest in resource-limited countries. In the last decade several large epidemics have led to a global push to improve and implement the tools for cholera prevention and control. Areas covered: PubMed, Google Scholar and the WHO website were searched to review the literature and summarize the current status of cholera vaccines to make recommendations on improving immunization approaches to cholera. Oral cholera vaccines (OCVs) have demonstrated their effectiveness in endemic, outbreak response and emergency settings, highlighting their potential for wider adoption. While two doses of the currently available OCVs are recommended by manufacturers, a single dose would be easier to implement. Encouragingly, recent studies have shown that cold chain requirements may no longer be essential. The establishment of the global OCV stockpile in 2013 has been a major advance in cholera preparedness. New killed and live-attenuated vaccines are being actively explored as candidate vaccines for endemic settings and/or as a traveller's vaccine. The recent advances in cholera vaccination approaches should be considered in the global cholera control strategy. Expert commentary: The development of affordable cholera vaccines is a major success to improve cholera control. New vaccines and country specific interventions will further reduce the burden of this disease globally.

  11. Selection of cholera toxin specific IgNAR single-domain antibodies from a naïve shark library.

    Science.gov (United States)

    Liu, Jinny L; Anderson, George P; Delehanty, James B; Baumann, Richard; Hayhurst, Andrew; Goldman, Ellen R

    2007-03-01

    Shark immunoglobulin new antigen receptor (IgNAR, also referred to as NAR) variable domains (Vs) are single-domain antibody (sdAb) fragments containing only two hypervariable loop structures forming 3D topologies for a wide range of antigen recognition and binding. Their small size ( approximately 12kDa) and high solubility, thermostability and binding specificity make IgNARs an exceptional alternative source of engineered antibodies for sensor applications. Here, two new shark NAR V display libraries containing >10(7) unique clones from non-immunized (naïve) adult spiny dogfish (Squalus acanthias) and smooth dogfish (Mustelus canis) sharks were constructed. The most conserved consensus sequences derived from random clone sequence were compared with published nurse shark (Ginglymostoma cirratum) sequences. Cholera toxin (CT) was chosen for panning one of the naïve display libraries due to its severe pathogenicity and commercial availability. Three very similar CT binders were selected and purified soluble monomeric anti-CT sdAbs were characterized using Luminex(100) and traditional ELISA assays. These novel anti-CT sdAbs selected from our newly constructed shark NAR V sdAb library specifically bound to soluble antigen, without cross reacting with other irrelevant antigens. They also showed superior heat stability, exhibiting slow loss of activity over the course of one hour at high temperature (95 degrees C), while conventional antibodies lost all activity in the first 5-10min. The successful isolation of target specific sdAbs from one of our non-biased NAR libraries, demonstrate their ability to provide binders against an unacquainted antigen of interest.

  12. Bacteriemia por Vibrio cholerae no-O1, no-O139 en un paciente en hemodiálisis crónica Non-O1, non-O139 Vibrio cholerae bacteremia in a chronic hemodialysis patient

    Directory of Open Access Journals (Sweden)

    Mariela S. Zárate

    2011-06-01

    Full Text Available Vibrio cholerae no-O1, no-O139 es un agente poco frecuente como causal de bacteriemias y no hay informes que documenten su presencia en pacientes en hemodiálisis crónica. Se describe el caso de una paciente en hemodiálisis crónica que presentó un cuadro de sepsis, por lo cual inició un tratamiento con vancomicina y ceftacidima. Al cabo de seis horas y media de incubación en el sistema BACT/ALERT de hemocultivo, se evidenció la presencia de bacilos curvos gram negativos, posteriormente identificados como Vibrio cholerae mediante pruebas bioquímicas convencionales y el uso de los kits API 20 NE y VITEK 2. La evaluación del serogrupo y de la presencia de factores de patogenicidad, realizada en el laboratorio de referencia, determinó que el microorganismo hallado pertenecía al serogrupo no-O1, no-O139. No se detectó la toxina de cólera, tampoco el factor de colonización ni la toxina termoestable. El aislamiento presentó sensibilidad frente a ampicilina, trimetoprima-sulfametoxazol, ciprofloxacina, tetraciclina, ceftacidima y cefotaxima por el método de difusión con discos y por VITEK 2. La paciente cumplió 14 días de tratamiento con ceftacidima endovenosa, con evolución favorable.Non-O1, and non-O139 Vibrio cholerae is an infrequent cause of bacteremia. There are no reports of such bacteremia in chronic hemodialysis patients. This work describes the case of a chronic hemodialysis patient that had an episode of septicemia associated with dialysis. Blood cultures were obtained and treatment was begun with vancomycin and ceftazidime. After 6.5 hours of incubation in the Bact/Alert system there is evidence of gram-negative curved bacilli that were identified as Vibrio cholerae by conventional biochemical tests, API 20 NE and the VITEK 2 system. This microorganism was sent to the reference laboratory for evaluation of serogroup and virulence factors and was identified as belonging to the non-O1 and non-O139 serogroup. The cholera

  13. Impact of oral cholera vaccines in cholera-endemic countries: A mathematical modeling study.

    Science.gov (United States)

    Kim, Jong-Hoon; Mogasale, Vittal; Burgess, Colleen; Wierzba, Thomas F

    2016-04-19

    Impact evaluation of vaccination programs is necessary for making decisions to introduce oral cholera vaccines (OCVs) in cholera-endemic countries. We analyzed data to forecast the future global burden of cholera. We developed a mathematical model of cholera transmission in three countries as examples: Nigeria, Uganda, and Indonesia. After fitting the model, we evaluated the impact of OCVs delivered in four vaccination strategies varying by target age group and frequency of vaccination over the period of 2015-2030. Data suggest that the global annual incidence of cholera will increase from 3046238 in 2015 to 3787385 in 2030 with the highest burden in Asia and Africa where overall population size is large and the proportion of population with access to improved sanitation facilities is low. We estimate that OCV will reduce the cumulative incidence of cholera by half in Indonesia and >80% in Nigeria and Uganda when delivered to 1+ year olds every three years at a coverage rate of 50%, although cholera may persist through higher coverage rates (i.e., >90%). The proportion of person-to-person transmission compared to water-to-person transmission is positively correlated with higher vaccination impact in all three countries. Periodic OCV vaccination every three or five years can significantly reduce the global burden of cholera although cholera may persist even with high OCV coverage. Vaccination impact will likely vary depending on local epidemiological conditions including age distribution of cases and relative contribution of different transmission routes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Effect of pertussis and cholera toxins administered supraspinally on CA3 hippocampal neuronal cell death and the blood glucose level induced by kainic acid in mice.

    Science.gov (United States)

    Kim, Chea-Ha; Park, Soo-Hyun; Sim, Yun-Beom; Sharma, Naveen; Kim, Sung-Su; Lim, Su-Min; Jung, Jun-Sub; Suh, Hong-Won

    2014-12-01

    The effect of cholera toxin (CTX) or pertussis toxin (PTX) administered supraspinally on hippocampal neuronal cell death in CA3 region induced by kainic acid (KA) was examined in mice. After the pretreatment with either PTX or CTX intracerebroventricularly (i.c.v.), mice were administered i.c.v. with KA. The i.c.v. treatment with KA caused a neuronal cell death in CA3 region and PTX, but not CTX, attenuated the KA-induced neuronal cell death. In addition, i.c.v. treatment with KA caused an elevation of the blood glucose level. The i.c.v. PTX pretreatment alone caused a hypoglycemia and inhibited KA-induced hyperglycemic effect. However, i.c.v. pretreatment with CTX did not affect the basal blood glucose level and KA-induced hyperglycemic effect. Moreover, KA administered i.c.v. caused an elevation of corticosterone level and reduction of the blood insulin level. Whereas, i.c.v. pretreatment with PTX further enhanced KA-induced up-regulation of corticosterone level. Furthermore, i.c.v. administration of PTX alone increased the insulin level and KA-induced hypoinsulinemic effect was reversed. In addition, PTX pretreatment reduces the KA-induced seizure activity. Our results suggest that supraspinally administered PTX, exerts neuroprotective effect against KA-induced neuronal cells death in CA3 region and neuroprotective effect of PTX is mediated by the reduction of KA-induced blood glucose level. Copyright © 2014 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

  15. Drinking cholera

    DEFF Research Database (Denmark)

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

    2015-01-01

    . cholerae can survive in the river systems in Bangladesh; however,water sources which have been contaminated with river water are avoided as potential drinkingwater sources. Furthermore, there are no physical connecting points between the river system anddrinking water sources among the study population......, indicating that the primary driver for choleracases in Bangladesh is likely not through the contamination of saline-rich river water into drinkingwater sources.......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...

  16. Independent Regulation of Type VI Secretion in Vibrio cholerae by TfoX and TfoY

    Directory of Open Access Journals (Sweden)

    Lisa C. Metzger

    2016-05-01

    Full Text Available Type VI secretion systems (T6SSs are nanomachines used for interbacterial killing and intoxication of eukaryotes. Although Vibrio cholerae is a model organism for structural studies on T6SSs, the underlying regulatory network is less understood. A recent study showed that the T6SS is part of the natural competence regulon in V. cholerae and is activated by the regulator TfoX. Here, we identify the TfoX homolog TfoY as a second activator of the T6SS. Importantly, despite inducing the same T6SS core machinery, the overall regulons differ significantly for TfoX and TfoY. We show that TfoY does not contribute to competence induction. Instead, TfoY drives the production of T6SS-dependent and T6SS-independent toxins, together with an increased motility phenotype. Hence, we conclude that V. cholerae uses its sole T6SS in response to diverse cues and for distinctive outcomes: either to kill for the prey’s DNA, leading to horizontal gene transfer, or as part of a defensive escape reaction.

  17. Promotion of Cholera Awareness Among Households of Cholera Patients: A Randomized Controlled Trial of the Cholera-Hospital-Based-Intervention-for-7 Days (CHoBI7) Intervention.

    Science.gov (United States)

    Saif-Ur-Rahman, K M; Parvin, Tahmina; Bhuyian, Sazzadul Islam; Zohura, Fatema; Begum, Farzana; Rashid, Mahamud-Ur; Biswas, Shwapon Kumar; Sack, David; Sack, R Bradley; Monira, Shirajum; Alam, Munirul; Shaly, Nusrat Jahan; George, Christine Marie

    2016-12-07

    Previous studies have demonstrated that household contacts of cholera patients are highly susceptible to cholera infections for a 7-day period after the presentation of the index patient in the hospital. However, there is no standard of care to prevent cholera transmission in this high-risk population. Furthermore, there is limited information available on awareness of cholera transmission and prevention among cholera patients and their household contacts. To initiate a standard of care for this high-risk population, we developed the Cholera-Hospital-Based-Intervention-for-7-Days (CHoBI7), which delivers a handwashing with soap and water treatment intervention to household contacts during the time they spend with the admitted cholera patient in the hospital and reinforces these messages through home visits. To test CHoBI7, we conducted a randomized controlled trial among 302 intervention cholera patient household members and 302 control cholera patient household members in Dhaka, Bangladesh. In this study, we evaluated the effectiveness of the CHoBI7 intervention in increasing awareness of cholera transmission and prevention, and the key times for handwashing with soap. We observed a significant increase in cholera knowledge score in the intervention arm compared with the control arm at both the 1-week follow-up {score coefficient = 2.34 (95% confidence interval [CI] = 1.96, 2.71)} and 6 to 12-month follow-up period (score coefficient = 1.59 [95% CI = 1.05, 2.13]). This 1-week hospital- and home-based intervention led to a significant increase in knowledge of cholera transmission and prevention which was sustained 6 to 12 months post-intervention. These findings suggest that the CHoBI7 intervention presents a promising approach to increase cholera awareness among this high-risk population. © The American Society of Tropical Medicine and Hygiene.

  18. Cholera - management and prevention.

    Science.gov (United States)

    Davies, Hannah G; Bowman, Conor; Luby, Stephen P

    2017-06-01

    Cholera is an acute secretory diarrhoeal infection caused by the bacterium Vibrio cholerae. It is likely to have originated in the Indian sub-continent; however, it spread to cause six worldwide pandemics between 1817-1923. The ongoing seventh worldwide pandemic of cholera began in 1961. The intensity, duration and severity of cholera epidemics have been increasing, signaling the need for more effective control and prevention measures. The response to the cholera pandemics of the 19th century led to the development of safe and effective sanitation and water systems which have effectively removed the risk of cholera in many settings. However, such systems are not in place to protect billions of people worldwide. Although some progress has been made in expanding access to water in recent years, achieving optimal infrastructure will, in the most optimistic scenario, take decades. Climate change, extreme weather events and rapid urbanisation suggests that alternatives to the current paradigm of providing large centralised water and sanitation systems should be considered, including smaller decentralised systems. The aim of this review paper is to provide an overview of current knowledge regarding management of cholera with a focus on prevention measures including vaccination and water and sanitation interventions. © 2017 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

    Choi, Seon Young; Rashed, Shah M; Hasan, Nur A; Alam, Munirul; Islam, Tarequl; Sadique, Abdus; Johura, Fatema-Tuz; Eppinger, Mark; Ravel, Jacques; Huq, Anwar; Cravioto, Alejandro; Colwell, Rita R

    2016-03-15

    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. Sequencing of genomes of V. cholerae is critical if genetic changes occurring over time in the circulating population of an area of endemicity are to be understood. Although cholera outbreaks occurred rarely

  20. Phase Partitioning of GM1 and Its Bodipy-Labeled Analog Determine Their Different Binding to Cholera Toxin

    Directory of Open Access Journals (Sweden)

    Sami Rissanen

    2017-05-01

    Full Text Available Driven by interactions between lipids and proteins, biological membranes display lateral heterogeneity that manifests itself in a mosaic of liquid-ordered (Lo or raft, and liquid-disordered (Ld or non-raft domains with a wide range of different properties and compositions. In giant plasma membrane vesicles and giant unilamellar vesicles, specific binding of Cholera Toxin (CTxB to GM1 glycolipids is a commonly used strategy to label raft domains or Lo membrane environments. However, these studies often use acyl-chain labeled bodipy-GM1 (bdGM1, whose headgroup accessibility and membrane order or phase partitioning may differ from those of GM1, rendering the interpretation of CTxB binding data quite problematic. To unravel the molecular basis of CTxB binding to GM1 and bdGM1, we explored the partitioning and the headgroup presentation of these gangliosides in the Lo and Ld phases using atomistic molecular dynamics simulations complemented by CTxB binding experiments. The conformation of both GM1 and bdGM1 was shown to be largely similar in the Lo and Ld phases. However, bdGM1 showed reduction in receptor availability when reconstituted into synthetic bilayer mixtures, highlighting that membrane phase partitioning of the gangliosides plays a considerable role in CTxB binding. Our results suggest that the CTxB binding is predominately modulated by the partitioning of the receptor to an appropriate membrane phase. Further, given that the Lo and Ld partitioning of bdGM1 differs from those of GM1, usage of bdGM1 for studying GM1 behavior in cells can lead to invalid interpretation of experimental data.

  1. Knowledge, Attitudes, and Practices regarding Diarrhea and Cholera following an Oral Cholera Vaccination Campaign in the Solomon Islands.

    Science.gov (United States)

    Burnett, Eleanor; Dalipanda, Tenneth; Ogaoga, Divi; Gaiofa, Jenny; Jilini, Gregory; Halpin, Alison; Dietz, Vance; Date, Kashmira; Mintz, Eric; Hyde, Terri; Wannemuehler, Kathleen; Yen, Catherine

    2016-08-01

    In response to a 2011 cholera outbreak in Papua New Guinea, the Government of the Solomon Islands initiated a cholera prevention program which included cholera disease prevention and treatment messaging, community meetings, and a pre-emptive cholera vaccination campaign targeting 11,000 children aged 1-15 years in selected communities in Choiseul and Western Provinces. We conducted a post-vaccination campaign, household-level survey about knowledge, attitudes, and practices regarding diarrhea and cholera in areas targeted and not targeted for cholera vaccination. Respondents in vaccinated areas were more likely to have received cholera education in the previous 6 months (33% v. 9%; p = 0.04), to know signs and symptoms (64% vs. 22%; p = 0.02) and treatment (96% vs. 50%; p = 0.02) of cholera, and to be aware of cholera vaccine (48% vs. 14%; p = 0.02). There were no differences in water, sanitation, and hygiene practices. This pre-emptive OCV campaign in a cholera-naïve community provided a unique opportunity to assess household-level knowledge, attitudes, and practices regarding diarrhea, cholera, and water, sanitation, and hygiene (WASH). Our findings suggest that education provided during the vaccination campaign may have reinforced earlier mass messaging about cholera and diarrheal disease in vaccinated communities.

  2. Knowledge, Attitudes, and Practices regarding Diarrhea and Cholera following an Oral Cholera Vaccination Campaign in the Solomon Islands.

    Directory of Open Access Journals (Sweden)

    Eleanor Burnett

    2016-08-01

    Full Text Available In response to a 2011 cholera outbreak in Papua New Guinea, the Government of the Solomon Islands initiated a cholera prevention program which included cholera disease prevention and treatment messaging, community meetings, and a pre-emptive cholera vaccination campaign targeting 11,000 children aged 1-15 years in selected communities in Choiseul and Western Provinces.We conducted a post-vaccination campaign, household-level survey about knowledge, attitudes, and practices regarding diarrhea and cholera in areas targeted and not targeted for cholera vaccination. Respondents in vaccinated areas were more likely to have received cholera education in the previous 6 months (33% v. 9%; p = 0.04, to know signs and symptoms (64% vs. 22%; p = 0.02 and treatment (96% vs. 50%; p = 0.02 of cholera, and to be aware of cholera vaccine (48% vs. 14%; p = 0.02. There were no differences in water, sanitation, and hygiene practices.This pre-emptive OCV campaign in a cholera-naïve community provided a unique opportunity to assess household-level knowledge, attitudes, and practices regarding diarrhea, cholera, and water, sanitation, and hygiene (WASH. Our findings suggest that education provided during the vaccination campaign may have reinforced earlier mass messaging about cholera and diarrheal disease in vaccinated communities.

  3. Cholera in Azov area

    Directory of Open Access Journals (Sweden)

    O. N. Domashenko

    2015-01-01

    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

  4. In a time of cholera.

    Science.gov (United States)

    Grace, P A

    2014-03-01

    Dr. Nathaniel Alcock in his book A treatise on cholera described 22 cases of cholera that he treated in 1832. Blood-letting, either by leeches or venesection, was an essential part of the treatment. The belief was that reducing the blood volume would relieve stress on the heart and lungs allowing for better function. The receipts of the Townsend Street Cholera Hospital where Dr. Alcock worked show how extensive the practice was. Outside Dublin, local Boards of Health dealt with the cholera epidemic. Various public measures such as street cleaning and removal of patients to temporary hospitals were undertaken and various cures were tried. The overall mortality rate from cholera in Ireland during the epidemic was 38 %, but in some areas much higher. Even as cholera was spreading in the 1830s, a number of doctors were showing that intravenous fluids could dramatically alter the course of the disease. Unfortunately, their work was ignored and blood-letting continued to be a major component of the treatment of cholera for another 55 years.

  5. Estrogen inhibits chloride secretion caused by cholera and Escherichia coli enterotoxins in female rat distal colon.

    OpenAIRE

    Alzamora, Rodrigo; O'Mahony, Fiona; Harvey, Brian J

    2011-01-01

    Excessive Cl(-) secretion is the driving force for secretory diarrhea. 17β-Estradiol has been shown to inhibit Cl(-) secretion in rat distal colon through a nongenomic pathway. We examined whether 17β-estradiol inhibits Cl(-) secretion in an animal model of secretory diarrhea and the downstream effectors involved. The effect of 17β-estradiol on cholera toxin and heat-stable enterotoxin induced Cl(-) secretion in rat colonic mucosal sheets was studied by current-voltage clamping. Selective per...

  6. Identifying cholera "hotspots" in Uganda: An analysis of cholera surveillance data from 2011 to 2016

    OpenAIRE

    Bwire, Godfrey; Ali, Mohammad; Sack, David A.; Nakinsige, Anne; Naigaga, Martha; Debes, Amanda K.; Ngwa, Moise C.; Brooks, W. Abdullah; Garimoi Orach, Christopher

    2017-01-01

    Background Despite advance in science and technology for prevention, detection and treatment of cholera, this infectious disease remains a major public health problem in many countries in sub-Saharan Africa, Uganda inclusive. The aim of this study was to identify cholera hotspots in Uganda to guide the development of a roadmap for prevention, control and elimination of cholera in the country. Methodology/Principle findings We obtained district level confirmed cholera outbreak data from 2011 t...

  7. What is cholera?

    DEFF Research Database (Denmark)

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

    2016-01-01

    more commonly described than death (47%) as negative effects of cholera. Conclusions: The results from this study are suggestive of a need for reformulation of cholera and diarrhea communication. Messaging should be based on signs of dehydration, foregoing the use of medical terminology....

  8. Pertussis toxin substrate is a guanosine 5'-[beta-thio]diphosphate-, N-ethylmaleimide-, Mg2+- and temperature-sensitive GTP-binding protein.

    OpenAIRE

    Wong, S K; Martin, B R; Tolkovsky, A M

    1985-01-01

    We compared the effects of guanine nucleotides and Mg2+ on ADP-ribosylation of rat brain and liver membrane proteins catalysed by Bordetella pertussis toxin (IAP) and cholera toxin (CT). Labelling of proteins in the presence of [alpha-32P]NAD+, ATP and CT required GTP or guanosine 5'-[gamma-thio]triphosphate (GTP [S]). In contrast, labelling of one (liver) or two (brain) polypeptides by IAP was enhanced by guanosine 5'-[beta-thio]diphosphate (GDP[S]) or GTP, but was blocked by GTP[S] or guano...

  9. Multi-site cholera surveillance within the African Cholera Surveillance Network shows endemicity in Mozambique, 2011-2015.

    Science.gov (United States)

    Semá Baltazar, Cynthia; Langa, José Paulo; Dengo Baloi, Liliana; Wood, Richard; Ouedraogo, Issaka; Njanpop-Lafourcade, Berthe-Marie; Inguane, Dorteia; Elias Chitio, Jucunu; Mhlanga, Themba; Gujral, Lorna; D Gessner, Bradford; Munier, Aline; A Mengel, Martin

    2017-10-01

    Mozambique suffers recurrent annual cholera outbreaks especially during the rainy season between October to March. The African Cholera Surveillance Network (Africhol) was implemented in Mozambique in 2011 to generate accurate detailed surveillance data to support appropriate interventions for cholera control and prevention in the country. Africhol was implemented in enhanced surveillance zones located in the provinces of Sofala (Beira), Zambézia (District Mocuba), and Cabo Delgado (Pemba City). Data were also analyzed from the three outbreak areas that experienced the greatest number of cases during the time period under observation (in the districts of Cuamba, Montepuez, and Nampula). Rectal swabs were collected from suspected cases for identification of Vibrio cholerae, as well as clinical, behavioral, and socio-demographic variables. We analyzed factors associated with confirmed, hospitalized, and fatal cholera using multivariate logistic regression models. A total of 1,863 suspected cases and 23 deaths (case fatality ratio (CFR), 1.2%) were reported from October 2011 to December 2015. Among these suspected cases, 52.2% were tested of which 23.5% were positive for Vibrio cholerae O1 Ogawa. Risk factors independently associated with the occurrence of confirmed cholera were living in Nampula city district, the year 2014, human immunodeficiency virus infection, and the primary water source for drinking. Cholera was endemic in Mozambique during the study period with a high CFR and identifiable risk factors. The study reinforces the importance of continued cholera surveillance, including a strong laboratory component. The results enhanced our understanding of the need to target priority areas and at-risk populations for interventions including oral cholera vaccine (OCV) use, and assess the impact of prevention and control strategies. Our data were instrumental in informing integrated prevention and control efforts during major cholera outbreaks in recent years.

  10. Effect of cholera toxin administered supraspinally or spinally on the blood glucose level in pain and d-glucose fed animal models.

    Science.gov (United States)

    Sim, Yun-Beom; Park, Soo-Hyun; Kang, Yu-Jung; Kim, Sung-Su; Kim, Chea-Ha; Kim, Su-Jin; Jung, Jun-Sub; Ryu, Ohk-Hyun; Choi, Moon-Gi; Choi, Seong-Soo; Suh, Hong-Won

    2013-04-01

    In the present study, the effect of intrathecal (i.t.) or intracerebroventricular (i.c.v.) administration with cholera toxin (CTX) on the blood glucose level was examined in ICR mice. The i.t. treatment with CTX alone for 24 h dose-dependently increased the blood glucose level. However, i.c.v. treatment with CTX for 24 h did not affect the blood glucose level. When mice were orally fed with D-glucose (2 g/kg), the blood glucose level reached to a maximum level at 30 min and almost returned to the control level at 120 min after D-glucose feeding. I.c.v. pretreatment with CTX increased the blood glucose level in a potentiative manner, whereas i.t. pretreatment with CTX increased the blood glucose level in an additive manner in a D-glucose fed group. In addition, the blood glucose level was increased in formalin-induced pain animal model. I.c.v. pretreatment with CTX enhanced the blood glucose level in a potentiative manner in formalin-induced pain animal model. On the other hand, i.t. pretreatment with CTX increased the blood glucose level in an additive manner in formalin-induced pain animal model. Our results suggest that CTX administered supraspinally or spinally differentially modulates the regulation of the blood glucose level in D-glucose fed model as well as in formalin-induced pain model.

  11. Environmental Monitoring of Endemic Cholera

    Science.gov (United States)

    ElNemr, W.; Jutla, A. S.; Constantin de Magny, G.; Hasan, N. A.; Islam, M.; Sack, R.; Huq, A.; Hashem, F.; Colwell, R.

    2012-12-01

    Cholera remains a major public health threat. Since Vibrio cholerae, the causative agent of the disease, is autochthonous to riverine, estuarine, and coastal waters, it is unlikely the bacteria can be eradicated from its natural habitat. Prediction of disease, in conjunction with preventive vaccination can reduce the prevalence rate of a disease. Understanding the influence of environmental parameters on growth and proliferation of bacteria is an essential first step in developing prediction methods for outbreaks. Large scale geophysical variables, such as SST and coastal chlorophyll, are often associated with conditions favoring growth of V. cholerae. However, local environmental factors, meaning biological activity in ponds from where the bulk of populations in endemic regions derive water for daily usage, are either neglected or oversimplified. Using data collected from several sites in two geographically distinct locations in South Asia, we have identified critical local environmental factors associated with cholera outbreak. Of 18 environmental variables monitored for water sources in Mathbaria (a coastal site near the Bay of Bengal) and Bakergonj (an inland site) of Bangladesh, water depth and chlorophyll were found to be important factors associated with initiation of cholera outbreaks. Cholera in coastal regions appears to be related to intrusion. However, monsoonal flooding creates conditions for cholera epidemics in inland regions. This may be one of the first attempts to relate in-situ environmental observations with cholera. We anticipate that it will be useful for further development of prediction models in the resource constrained regions.

  12. [Microbiological characterization of non-O1 Vibrio cholerae isolated in Cuba].

    Science.gov (United States)

    Bravo Fariñas, Laura; Fernández, Anabel; Ramírez, María M; Llop, Alina; Martínez, Gerardo; Hernández, Raquel I; Cabrera, Luis E; Morier, Luis; Fraga, Jorge; Núñez, Fidel A; Aguila, Adalberto

    2007-01-01

    The study of 422 non-01 Vibrio cholerae strains from nine provinces, 9 of them isolated from a water-borne disease outbreak, was performed. All the strains exhibited antimicrobial susceptibility and virulence factors. The nine strains from the outbreak were subjected to a DNA macrorestriction study based on the pulsed field electrophoresis technique. For the first time in Cuba and the Caribbean. The circulation of atypical non-01 V cholerae strains (resistent to vibriostatic compound 0129 and trimethoprim/sulfamethoxazole). The behavior of antimicrobial susceptibility evinced for the first time the circulation of two different resistence patterns in Cuba (ampicilline, trimethoprim/ sulfamethoxazole, sulfonamide and tetracycline, trimethoprim/ sulfamethoxazole, sulfonamide). The frequency of trimethoprim/ sulfamethoxazole-resistent strains was similar during the whole period of study. However, resistance to ampicilline decreased whereas resistance to tetracycline increased. The main found virulence factors were gelatinase, hemolysine, elastase and adherence to Hep-2 cells. On the other hand, the outbreak strains showed higher percentages than the others due to the presence of heat-liable toxin and fimbriae. The results of the molecular and epidemiological studies allowed giving a speedy and accurate response that explained the etiology of the first food-borne disease outbreak.

  13. Antibiotics resistance in El Tor Vibrio cholerae 01 isolated during cholera outbreaks in Mozambique from 2012 to 2015.

    Directory of Open Access Journals (Sweden)

    Liliana Candida Dengo-Baloi

    Full Text Available Mozambique has recorded cyclically epidemic outbreaks of cholera. Antibiotic therapy is recommended in specific situations for management and control of cholera outbreaks. However, an increase in resistance rates to antibiotics by Vibrio cholerae has been reported in several epidemic outbreaks worldwide. On the other hand, there are few recent records of continuous surveillance of antibiotics susceptibility pattern of V. cholerae in Mozambique.The purpose of this study was to evaluate antibiotics resistance pattern of Vibrio cholerae O1 Ogawa isolated during Cholera outbreaks in Mozambique to commonly used antibiotics.We analyzed data from samples received in the context of surveillance and response to Cholera outbreaks in the National Reference Laboratory of Microbiology from the National Institute of Health of Mozambique, 159 samples suspected of cholera from cholera treatment centers of, Metangula (09, Memba (01, Tete City (08, Moatize (01, Morrumbala (01 districts, City of Quelimane (01, Lichinga (06 and Nampula (86 districts, from 2012 to 2015. Laboratory culture and standard biochemical tests were employed to isolate and identify Vibrio cholerae; serotypes were determined by antisera agglutination reaction in blade. Biotype and presence of important virulence factors analysis was done by PCR. Antibiotics susceptibility pattern was detected by disk diffusion method Kirby Bauer. Antibiotic susceptibility and results were interpreted by following as per recommendations of CLSI (Clinical and Laboratory Standards Institute 2014. All samples were collected and tested in the context of Africhol Project, approved by the National Bioethics Committee for Health.Among isolates from of Vibrio cholerae O1 El Tor Ogawa resistance to Sulphamethoxazole-trimethropim was 100% (53/53 to Trimethoprim-, being 100% (54/54 for Ampicillin, 99% (72/74 for Nalidixic Acid, 97% (64/66 to Chloramphenicol, 95% (42/44 for Nitrofurantoin and (19/20 Cotrimoxazole, 83% (80

  14. Antibiotics resistance in El Tor Vibrio cholerae 01 isolated during cholera outbreaks in Mozambique from 2012 to 2015.

    Science.gov (United States)

    Dengo-Baloi, Liliana Candida; Semá-Baltazar, Cynthia Amino; Manhique, Lena Vania; Chitio, Jucunu Elias; Inguane, Dorteia Luísa; Langa, José Paulo

    2017-01-01

    Mozambique has recorded cyclically epidemic outbreaks of cholera. Antibiotic therapy is recommended in specific situations for management and control of cholera outbreaks. However, an increase in resistance rates to antibiotics by Vibrio cholerae has been reported in several epidemic outbreaks worldwide. On the other hand, there are few recent records of continuous surveillance of antibiotics susceptibility pattern of V. cholerae in Mozambique. The purpose of this study was to evaluate antibiotics resistance pattern of Vibrio cholerae O1 Ogawa isolated during Cholera outbreaks in Mozambique to commonly used antibiotics. We analyzed data from samples received in the context of surveillance and response to Cholera outbreaks in the National Reference Laboratory of Microbiology from the National Institute of Health of Mozambique, 159 samples suspected of cholera from cholera treatment centers of, Metangula (09), Memba (01), Tete City (08), Moatize (01), Morrumbala (01) districts, City of Quelimane (01), Lichinga (06) and Nampula (86) districts, from 2012 to 2015. Laboratory culture and standard biochemical tests were employed to isolate and identify Vibrio cholerae; serotypes were determined by antisera agglutination reaction in blade. Biotype and presence of important virulence factors analysis was done by PCR. Antibiotics susceptibility pattern was detected by disk diffusion method Kirby Bauer. Antibiotic susceptibility and results were interpreted by following as per recommendations of CLSI (Clinical and Laboratory Standards Institute) 2014. All samples were collected and tested in the context of Africhol Project, approved by the National Bioethics Committee for Health. Among isolates from of Vibrio cholerae O1 El Tor Ogawa resistance to Sulphamethoxazole-trimethropim was 100% (53/53) to Trimethoprim-, being 100% (54/54) for Ampicillin, 99% (72/74) for Nalidixic Acid, 97% (64/66) to Chloramphenicol, 95% (42/44) for Nitrofurantoin and (19/20) Cotrimoxazole, 83% (80

  15. Cholera Fact Sheet

    Science.gov (United States)

    ... news-room/fact-sheets/detail/cholera","@context":"http://schema.org","@type":"Article"}; العربية 中文 français русский español ... that includes feedback at the local level and information-sharing at the global level. Cholera cases are ...

  16. Luminal cholera toxin alters motility in isolated guinea-pig jejunum via a pathway independent of 5-HT3 receptors

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    Candice eFung

    2010-09-01

    Full Text Available Cholera toxin (CT is well established to produce diarrhoea by producing hyperactivity of the enteric neural circuits that regulate water and electrolyte secretion. Its effects on intestinal motor patterns are less well understood. We examined the effects of luminal CT on motor activity of guinea-pig jejunum in vitro. Segments of jejunum were cannulated at either end and mounted horizontally.Their contractile activity was video-imaged and the recordings were used to construct spatiotemporal maps of contractile activity with CT (1.25 μg/ml or 12.5 μg/ml in the lumen. Both concentrations of CT induced propulsive motor activity in jejunal segments. The effect of 1.25 μg/ml CT was markedly enhanced by coincubation with granisetron (5-HT3 antagonist, 1 μM, which prevents the hypersecretion induced by CT. The increased propulsive activity was not accompanied by increased segmentation and occurred very early after exposure to CT in the presence of granisetron. Luminal CT also reduced the pressure threshold for saline distension evoked propulsive reflexes, an effect resistant to granisetron. In contrast, CT prevented the induction of segmenting contractions by luminal decanoic acid, so its effects on propulsive and segmenting contractile activity are distinctly different. Thus, in addition to producing hypersecretion, CT excites propulsive motor activity with an entirely different time course and pharmacology, but inhibits nutrient induced segmentation. This suggests that CT excites more than one enteric neural circuit and that propulsive and segmenting motor patterns are differentially regulated.

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

    Directory of Open Access Journals (Sweden)

    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

  18. Cholera in the Americas.

    Science.gov (United States)

    1991-01-01

    The cholera epidemic 1st hit South America in January 1991 in the coastal town of Chancay, Peru. In 2 weeks, it spread over 2000 km of the Pacific coast. By the end of the 1st month, it had already reached the mountains and tropical forests. By August 1991, cholera cases were reported in order of appearances in Ecuador, Colombia, Chile, Brazil, the US, Mexico, Guatemala, Bolivia, and El Salvador. Health authorities still do not know how it was introduced into South America. The case fatality rate has remained at a low of 1%, probably due to the prompt actions of health authorities in informing the public of the epidemic and what preventive cautions should be taken. This epidemic is part of the 7th pandemic which originated in Celebes, Indonesia in 1961. Cholera can spread relatively unchecked in Latin America because sewage in urban areas is not treated even though they do have sewage collection systems. The untreated wastewater enters rivers and the ocean. Consumption of raw seafood is not unusual and has been responsible for cholera infection in some cases. In fact, many countries placed import restrictions on marine products from Peru following the outbreak at a loss of $US10-$US40 million. Municipal sewage treatment facilities, especially stabilization ponds, would prevent the spread of cholera and other pathogens. In rural areas, pit latrines located away from wells can effectively dispose of human wastes. Most water supplies in Latin America are not disinfected. Disinfection drinking water with adequate levels of chlorine would effectively destroy V. cholera. If this is not possible, boiling the water for 2-3 minutes would destroy the pathogen. Any cases of cholera must be reported to PAHO. PAHO has responded to the outbreak by forming a Cholera Task Force and arranged transport of oral rehydration salts, intravenous fluids, antibiotics, and other essential medical supplies.

  19. 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.

  20. Multi-site cholera surveillance within the African Cholera Surveillance Network shows endemicity in Mozambique, 2011–2015

    Science.gov (United States)

    Langa, José Paulo; Dengo Baloi, Liliana; Wood, Richard; Ouedraogo, Issaka; Njanpop-Lafourcade, Berthe-Marie; Inguane, Dorteia; Elias Chitio, Jucunu; Mhlanga, Themba; Gujral, Lorna; D. Gessner, Bradford; Munier, Aline; A. Mengel, Martin

    2017-01-01

    Background Mozambique suffers recurrent annual cholera outbreaks especially during the rainy season between October to March. The African Cholera Surveillance Network (Africhol) was implemented in Mozambique in 2011 to generate accurate detailed surveillance data to support appropriate interventions for cholera control and prevention in the country. Methodology/Principal findings Africhol was implemented in enhanced surveillance zones located in the provinces of Sofala (Beira), Zambézia (District Mocuba), and Cabo Delgado (Pemba City). Data were also analyzed from the three outbreak areas that experienced the greatest number of cases during the time period under observation (in the districts of Cuamba, Montepuez, and Nampula). Rectal swabs were collected from suspected cases for identification of Vibrio cholerae, as well as clinical, behavioral, and socio-demographic variables. We analyzed factors associated with confirmed, hospitalized, and fatal cholera using multivariate logistic regression models. A total of 1,863 suspected cases and 23 deaths (case fatality ratio (CFR), 1.2%) were reported from October 2011 to December 2015. Among these suspected cases, 52.2% were tested of which 23.5% were positive for Vibrio cholerae O1 Ogawa. Risk factors independently associated with the occurrence of confirmed cholera were living in Nampula city district, the year 2014, human immunodeficiency virus infection, and the primary water source for drinking. Conclusions/Significance Cholera was endemic in Mozambique during the study period with a high CFR and identifiable risk factors. The study reinforces the importance of continued cholera surveillance, including a strong laboratory component. The results enhanced our understanding of the need to target priority areas and at-risk populations for interventions including oral cholera vaccine (OCV) use, and assess the impact of prevention and control strategies. Our data were instrumental in informing integrated prevention and

  1. The cholera

    International Nuclear Information System (INIS)

    Castaneda, Elizabeth; De la Hoz, Fernando

    1996-01-01

    The cholera is a sharp intestinal infection, characterized by the abrupt appearance of abundant watery diarrhea and vomit, severe dehydration, metabolic imbalance and circulatory collapse; although it can study a symptomatic or to cause light symptomatology. In the cases serious non-treaties the death can take place in the 24 later hours to its appearance; the lethality in these cases can exceed 50 percent. However. With the treatment of re-hydration appropriate. This decreases to less ten percent. The cholera is related in direct form with the poverty; it is presented where accumulation and lack of measures of hygiene exist and of drinkable water. It is for it that the best strategy to prevent the illness is to eliminate the factors that favor its transmission: to improve the quality of the water and to implement elementary measures of hygiene. When the environmental conditions are favorable, the cholera has been absent

  2. Treatment with Entinostat Heals Experimental Cholera by Affecting Physical and Chemical Barrier Functions of Intestinal Epithelia.

    Science.gov (United States)

    Sarker, Protim; Banik, Atanu; Stromberg, Roger; Gudmundsson, Gudmundur H; Raqib, Rubhana; Agerberth, Birgitta

    2017-07-01

    We have shown previously that oral treatment with sodium butyrate or phenylbutyrate in an experimental model of shigellosis improves clinical outcomes and induces the expression of the antimicrobial peptide CAP-18 in the large intestinal epithelia. In a subsequent study, we found that entinostat, an aroylated phenylenediamine compound, has similar therapeutic potential against shigellosis. In this study, we aimed to evaluate entinostat as a potential candidate for host-directed therapy against cholera in an experimental model. Vibrio cholerae -infected rabbits were treated with two different dose regimens of entinostat: either 0.5 mg twice daily for 2 days or 1 mg once daily for 2 days. The effects of treatment on clinical outcomes and V. cholerae shedding (CFU count in stool) were observed. Immunohistochemical analysis was carried out to assess CAP-18 expression in ileal and jejunal mucosae. The serum zonulin level was measured by an enzyme-linked immunosorbent assay (ELISA) to evaluate gut permeability. Infection of rabbits with V. cholerae downregulated CAP-18 expression in the ileal epithelium; the expression was replenished by oral treatment with entinostat at either dose regimen. The level of zonulin, a marker of gut permeability, in serum was upregulated after infection, and this upregulation was counteracted after treatment with entinostat. Entinostat treatment also led to recovery from cholera and a decline in the V. cholerae count in stool. In conclusion, the improved clinical outcome of cholera for rabbits treated with entinostat is associated with the induction of CAP-18 and the reduction of gut epithelial permeability. Copyright © 2017 American Society for Microbiology.

  3. Cholera Prevention and Control

    Science.gov (United States)

    ... name=”commit” type=”submit” value=”Submit” /> Prevention & Control Recommend on Facebook Tweet Share Compartir Prevention of ... of cholera and other diarrheal disease prevention. Prevention & Control Topics Ending Cholera: The Global Roadmap to 2030 ...

  4. Follicle-stimulating hormone receptor-mediated uptake of 45Ca2+ by cultured rat Sertoli cells does not require activation of cholera toxin- or pertussis toxin-sensitive guanine nucleotide binding proteins or adenylate cyclase

    International Nuclear Information System (INIS)

    Grasso, P.; Reichert, L.E. Jr.

    1990-01-01

    We have previously reported that FSH stimulates flux of 45Ca2+ into cultured Sertoli cells from immature rats via voltage-sensitive and voltage-independent calcium channels. In the present study, we show that this effect of FSH does not require cholera toxin (CT)- or pertussis toxin (PT)-sensitive guanine nucleotide binding (G) protein or activation of adenylate cyclase (AC). Significant stimulation of 45Ca2+ influx was observed within 1 min, and maximal response (3.2-fold over basal levels) was achieved within 2 min after exposure to FSH. FSH-stimulated elevations in cellular cAMP paralleled increases in 45Ca2+ uptake, suggesting a possible coupling of AC activation to 45Ca2+ influx. (Bu)2cAMP, however, was not able to enhance 45Ca2+ uptake over basal levels at a final concentration of 1000 microM, although a concentration-related increase in androstenedione conversion to estradiol was evident. Exposure of Sertoli cells to CT (10 ng/ml) consistently stimulated basal levels of androstenedione conversion to estradiol but had no effect on basal levels of 45Ca2+ uptake. Similarly, CT had no effect on FSH-induced 45Ca2+ uptake, but potentiated FSH-stimulated estradiol synthesis. PT (10 ng/ml) augmented basal and FSH-stimulated estradiol secretion without affecting 45Ca2+ influx. The adenosine analog N6-phenylisopropyladenosine, which binds to Gi-coupled adenosine receptors on Sertoli cells, inhibited FSH-stimulated androgen conversion to estradiol in a dose-related (1-1000 nM) manner, but FSH-stimulated 45Ca2+ influx remained unchanged. Our results show that in contrast to FSH-stimulated estradiol synthesis, the flux of 45Ca2+ into Sertoli cells in response to FSH is not mediated either directly or indirectly by CT- or PT-sensitive G protein, nor does it require activation of AC. Our data further suggest that the FSH receptor itself may function as a calcium channel

  5. Cholera Illness and Symptoms

    Science.gov (United States)

    ... Share Compartir Cholera is an acute, diarrheal illness caused by infection of the intestine with the bacterium Vibrio cholerae and is spread by ingestion of contaminated food or water. The infection is often mild or without symptoms, ...

  6. The mucosal adjuvant cholera toxin B instructs non-mucosal dendritic cells to promote IgA production via retinoic acid and TGF-β.

    Directory of Open Access Journals (Sweden)

    Anouk K Gloudemans

    Full Text Available It is currently unknown how mucosal adjuvants cause induction of secretory immunoglobulin A (IgA, and how T cell-dependent (TD or -independent (TI pathways might be involved. Mucosal dendritic cells (DCs are the primary antigen presenting cells driving TI IgA synthesis, by producing a proliferation-inducing ligand (APRIL, B cell activating factor (BAFF, Retinoic Acid (RA, TGF-β or nitric oxide (NO. We hypothesized that the mucosal adjuvant Cholera Toxin subunit B (CTB could imprint non-mucosal DCs to induce IgA synthesis, and studied the mechanism of its induction. In vitro, CTB-treated bone marrow derived DCs primed for IgA production by B cells without the help of T cells, yet required co-signaling by different Toll-like receptor (TLR ligands acting via the MyD88 pathway. CTB-DC induced IgA production was blocked in vitro or in vivo when RA receptor antagonist, TGF-β signaling inhibitor or neutralizing anti-TGF-β was added, demonstrating the involvement of RA and TGF-β in promoting IgA responses. There was no major involvement for BAFF, APRIL or NO. This study highlights that synergism between CTB and MyD88-dependent TLR signals selectively imprints a TI IgA-inducing capacity in non-mucosal DCs, explaining how CTB acts as an IgA promoting adjuvant.

  7. The mucosal adjuvant cholera toxin B instructs non-mucosal dendritic cells to promote IgA production via retinoic acid and TGF-β.

    Science.gov (United States)

    Gloudemans, Anouk K; Plantinga, Maud; Guilliams, Martin; Willart, Monique A; Ozir-Fazalalikhan, Arifa; van der Ham, Alwin; Boon, Louis; Harris, Nicola L; Hammad, Hamida; Hoogsteden, Henk C; Yazdanbakhsh, Maria; Hendriks, Rudi W; Lambrecht, Bart N; Smits, Hermelijn H

    2013-01-01

    It is currently unknown how mucosal adjuvants cause induction of secretory immunoglobulin A (IgA), and how T cell-dependent (TD) or -independent (TI) pathways might be involved. Mucosal dendritic cells (DCs) are the primary antigen presenting cells driving TI IgA synthesis, by producing a proliferation-inducing ligand (APRIL), B cell activating factor (BAFF), Retinoic Acid (RA), TGF-β or nitric oxide (NO). We hypothesized that the mucosal adjuvant Cholera Toxin subunit B (CTB) could imprint non-mucosal DCs to induce IgA synthesis, and studied the mechanism of its induction. In vitro, CTB-treated bone marrow derived DCs primed for IgA production by B cells without the help of T cells, yet required co-signaling by different Toll-like receptor (TLR) ligands acting via the MyD88 pathway. CTB-DC induced IgA production was blocked in vitro or in vivo when RA receptor antagonist, TGF-β signaling inhibitor or neutralizing anti-TGF-β was added, demonstrating the involvement of RA and TGF-β in promoting IgA responses. There was no major involvement for BAFF, APRIL or NO. This study highlights that synergism between CTB and MyD88-dependent TLR signals selectively imprints a TI IgA-inducing capacity in non-mucosal DCs, explaining how CTB acts as an IgA promoting adjuvant.

  8. Cholera Toxin Induces Sustained Hyperexcitability in Myenteric, but Not Submucosal, AH Neurons in Guinea Pig Jejunum

    Directory of Open Access Journals (Sweden)

    Joel C. Bornstein

    2017-04-01

    Full Text Available Background and Aims: Cholera toxin (CT-induced hypersecretion requires activation of secretomotor pathways in the enteric nervous system (ENS. AH neurons, which have been identified as a population of intrinsic sensory neurons (ISNs, are a source of excitatory input to the secretomotor pathways. We therefore examined effects of CT in the intestinal lumen on myenteric and submucosal AH neurons.Methods: Isolated segments of guinea pig jejunum were incubated for 90 min with saline plus CT (12.5 μg/ml or CT + neurotransmitter antagonist, or CT + tetrodotoxin (TTX in their lumen. After washing CT away, submucosal or myenteric plexus preparations were dissected keeping circumferentially adjacent mucosa intact. Submucosal AH neurons were impaled adjacent to intact mucosa and myenteric AH neurons were impaled adjacent to, more than 5 mm from, and in the absence of intact mucosa. Neuronal excitability was monitored by injecting 500 ms current pulses through the recording electrode.Results: After CT pre-treatment, excitability of myenteric AH neurons adjacent to intact mucosa (n = 29 was greater than that of control neurons (n = 24, but submucosal AH neurons (n = 33, control n = 27 were unaffected. CT also induced excitability increases in myenteric AH neurons impaled distant from the mucosa (n = 6 or in its absence (n = 5. Coincubation with tetrodotoxin or SR142801 (NK3 receptor antagonist, but not SR140333 (NK1 antagonist or granisetron (5-HT3 receptor antagonist prevented the increased excitability induced by CT. Increased excitability was associated with a reduction in the characteristic AHP and an increase in the ADP of these neurons, but not a change in the hyperpolarization-activated inward current, Ih.Conclusions: CT increases excitability of myenteric, but not submucosal, AH neurons. This is neurally mediated and depends on NK3, but not 5-HT3 receptors. Therefore, CT may act to amplify the secretomotor response to CT via an increase in the

  9. Agent-based modelling of cholera diffusion

    NARCIS (Netherlands)

    Augustijn-Beckers, Petronella; Doldersum, Tom; Useya, Juliana; Augustijn, Dionysius C.M.

    2016-01-01

    This paper introduces a spatially explicit agent-based simulation model for micro-scale cholera diffusion. The model simulates both an environmental reservoir of naturally occurring V.cholerae bacteria and hyperinfectious V. cholerae. Objective of the research is to test if runoff from open refuse

  10. 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.

  11. Environmental Factors Influencing Epidemic Cholera

    Science.gov (United States)

    Jutla, Antarpreet; Whitcombe, Elizabeth; Hasan, Nur; Haley, Bradd; Akanda, Ali; Huq, Anwar; Alam, Munir; Sack, R. Bradley; Colwell, Rita

    2013-01-01

    Cholera outbreak following the earthquake of 2010 in Haiti has reaffirmed that the disease is a major public health threat. Vibrio cholerae is autochthonous to aquatic environment, hence, it cannot be eradicated but hydroclimatology-based prediction and prevention is an achievable goal. Using data from the 1800s, we describe uniqueness in seasonality and mechanism of occurrence of cholera in the epidemic regions of Asia and Latin America. Epidemic regions are located near regional rivers and are characterized by sporadic outbreaks, which are likely to be initiated during episodes of prevailing warm air temperature with low river flows, creating favorable environmental conditions for growth of cholera bacteria. Heavy rainfall, through inundation or breakdown of sanitary infrastructure, accelerates interaction between contaminated water and human activities, resulting in an epidemic. This causal mechanism is markedly different from endemic cholera where tidal intrusion of seawater carrying bacteria from estuary to inland regions, results in outbreaks. PMID:23897993

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

    Science.gov (United States)

    Boucher, Yan

    2016-05-03

    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. Copyright © 2016 Boucher.

  13. 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.

  14. How Will Climate Change Impact Cholera Outbreaks?

    Science.gov (United States)

    Nasr Azadani, F.; Jutla, A.; Rahimikolu, J.; Akanda, A. S.; Huq, A.; Colwell, R. R.

    2014-12-01

    Environmental parameters associated with cholera are well documented. However, cholera continues to be a global public health threat. Uncertainty in defining environmental processes affecting growth and multiplication of the cholera bacteria can be affected significantly by changing climate at different temporal and spatial scales, either through amplification of the hydroclimatic cycle or by enhanced variability of large scale geophysical processes. Endemic cholera in the Bengal Delta region of South Asia has a unique pattern of two seasonal peaks and there are associated with asymmetric and episodic variability in river discharge. The first cholera outbreak in spring is related with intrusion of bacteria laden coastal seawater during low river discharge. Cholera occurring during the fall season is hypothesized to be associated with high river discharge related to a cross-contamination of water resources and, therefore, a second wave of disease, a phenomenon characteristic primarily in the inland regions. Because of difficulties in establishing linkage between coarse resolutions of the Global Climate Model (GCM) output and localized disease outbreaks, the impact of climate change on diarrheal disease has not been explored. Here using the downscaling method of Support Vector Machines from HADCM3 and ECHAM models, we show how cholera outbreak patterns are changing in the Bengal Delta. Our preliminary results indicate statistically significant changes in both seasonality and magnitude in the occurrence of cholera over the next century. Endemic cholera is likely to transform into epidemic forms and new geographical areas will be at risk for cholera outbreaks.

  15. Vaxchora: A Single-Dose Oral Cholera Vaccine.

    Science.gov (United States)

    Cabrera, Adriana; Lepage, Jayne E; Sullivan, Karyn M; Seed, Sheila M

    2017-07-01

    To review trials evaluating the efficacy and safety of Vaxchora, a reformulated, single-dose, oral, lyophilized Vibrio cholerae CVD 103-HgR vaccine for the prevention of travel-related cholera caused by V cholerae serogroup O1. A literature search was conducted using MEDLINE (1946 to January week 3, 2017) and EMBASE (1996 to 2017 week 3). Keywords included oral cholera vaccine, single-dose, Vaxchora, and CVD 103-HgR. Limits included human, clinical trials published in English since 2010. ClinicalTrials.gov was used as a source for unpublished data. Additional data sources were obtained through bibliographic review of selected articles. Studies that addressed the safety and efficacy of Vaxchora, the reformulated, single-dose oral CVD 103-HgR cholera vaccine, were selected for analysis. Approval of Vaxchora, was based on efficacy of the vaccine in human trials demonstrating 90.3% protection among those challenged with V cholerae 10 days after vaccination and in immunogenicity studies with 90% systemic vibriocidal antibody conversion at 6 months after a single-dose of vaccine. Tolerability was acceptable, with the most common adverse effects reported to be fatigue, headache, and abdominal pain. Vaxchora is the only FDA-approved, single-dose oral vaccine for the prevention of cholera caused by V cholerae serogroup O1 in adult travelers from the United States going to cholera-affected areas. Safety and efficacy has not been established in children, immunocompromised persons, and pregnant or breastfeeding women or those living in cholera-endemic areas.

  16. [Complications and treatment of cholera during pregnancy].

    Science.gov (United States)

    Figueroa Damian, R; Villagrana Zesati, R; Kasis Ariceaga, D

    1994-07-01

    Since 1961 cholera has spread in many countries reaching a pandemic form. Since 1991 Mexico has been involved in this pandemia. Near 20% of all cases of cholera in our country happen in fertile women, so the possibility of the association between cholera and pregnancy is high. We present the case of a pregnant woman, who during her third trimester presented a episode of cholera, developing premature labor. Furthermore is revised the medical literature about the general principles of the management of cholera, and the association between pregnancy and the intestinal infection.

  17. Knowledge, Attitudes, and Practices regarding Diarrhea and Cholera following an Oral Cholera Vaccination Campaign in the Solomon Islands

    OpenAIRE

    Burnett, Eleanor; Dalipanda, Tenneth; Ogaoga, Divi; Gaiofa, Jenny; Jilini, Gregory; Halpin, Alison; Dietz, Vance; Date, Kashmira; Mintz, Eric; Hyde, Terri; Wannemuehler, Kathleen; Yen, Catherine

    2016-01-01

    Background In response to a 2011 cholera outbreak in Papua New Guinea, the Government of the Solomon Islands initiated a cholera prevention program which included cholera disease prevention and treatment messaging, community meetings, and a pre-emptive cholera vaccination campaign targeting 11,000 children aged 1?15 years in selected communities in Choiseul and Western Provinces. Methodology and Principal Findings We conducted a post-vaccination campaign, household-level survey about knowledg...

  18. Induction of indoleamine 2, 3-dioxygenase in human dendritic cells by a cholera toxin B subunit-proinsulin vaccine.

    Directory of Open Access Journals (Sweden)

    Jacques C Mbongue

    Full Text Available Dendritic cells (DC interact with naïve T cells to regulate the delicate balance between immunity and tolerance required to maintain immunological homeostasis. In this study, immature human dendritic cells (iDC were inoculated with a chimeric fusion protein vaccine containing the pancreatic β-cell auto-antigen proinsulin linked to a mucosal adjuvant the cholera toxin B subunit (CTB-INS. Proteomic analysis of vaccine inoculated DCs revealed strong up-regulation of the tryptophan catabolic enzyme indoleamine 2, 3-dioxygenase (IDO1. Increased biosynthesis of the immunosuppressive enzyme was detected in DCs inoculated with the CTB-INS fusion protein but not in DCs inoculated with proinsulin, CTB, or an unlinked combination of the two proteins. Immunoblot and PCR analyses of vaccine treated DCs detected IDO1mRNA by 3 hours and IDO1 protein synthesis by 6 hours after vaccine inoculation. Determination of IDO1 activity in vaccinated DCs by measurement of tryptophan degradation products (kynurenines showed increased tryptophan cleavage into N-formyl kynurenine. Vaccination did not interfere with monocytes differentiation into DC, suggesting the vaccine can function safely in the human immune system. Treatment of vaccinated DCs with pharmacological NF-κB inhibitors ACHP or DHMEQ significantly inhibited IDO1 biosynthesis, suggesting a role for NF-κB signaling in vaccine up-regulation of dendritic cell IDO1. Heat map analysis of the proteomic data revealed an overall down-regulation of vaccinated DC functions, suggesting vaccine suppression of DC maturation. Together, our experimental data indicate that CTB-INS vaccine induction of IDO1 biosynthesis in human DCs may result in the inhibition of DC maturation generating a durable state of immunological tolerance. Understanding how CTB-INS modulates IDO1 activity in human DCs will facilitate vaccine efficacy and safety, moving this immunosuppressive strategy closer to clinical applications for prevention

  19. Genetic Markers for Western Corn Rootworm Resistance to Bt Toxin

    OpenAIRE

    Flagel, Lex E.; Swarup, Shilpa; Chen, Mao; Bauer, Christopher; Wanjugi, Humphrey; Carroll, Matthew; Hill, Patrick; Tuscan, Meghan; Bansal, Raman; Flannagan, Ronald; Clark, Thomas L.; Michel, Andrew P.; Head, Graham P.; Goldman, Barry S.

    2015-01-01

    Western corn rootworm (WCR) is a major maize (Zea mays L.) pest leading to annual economic losses of more than 1 billion dollars in the United States. Transgenic maize expressing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are widely used for the management of WCR. However, cultivation of Bt-expressing maize places intense selection pressure on pest populations to evolve resistance. Instances of resistance to Bt toxins have been reported in WCR. Developing genet...

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

    International Nuclear Information System (INIS)

    Ahmed, S.

    2015-01-01

    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)

  1. Inhibition of cAMP-activated intestinal chloride secretion by diclofenac: cellular mechanism and potential application in cholera.

    Science.gov (United States)

    Pongkorpsakol, Pawin; Pathomthongtaweechai, Nutthapoom; Srimanote, Potjanee; Soodvilai, Sunhapas; Chatsudthipong, Varanuj; Muanprasat, Chatchai

    2014-09-01

    Cyclic AMP-activated intestinal Cl- secretion plays an important role in pathogenesis of cholera. This study aimed to investigate the effect of diclofenac on cAMP-activated Cl- secretion, its underlying mechanisms, and possible application in the treatment of cholera. Diclofenac inhibited cAMP-activated Cl- secretion in human intestinal epithelial (T84) cells with IC50 of ∼ 20 µM. The effect required no cytochrome P450 enzyme-mediated metabolic activation. Interestingly, exposures of T84 cell monolayers to diclofenac, either in apical or basolateral solutions, produced similar degree of inhibitions. Analyses of the apical Cl- current showed that diclofenac reversibly inhibited CFTR Cl- channel activity (IC50 ∼ 10 µM) via mechanisms not involving either changes in intracellular cAMP levels or CFTR channel inactivation by AMP-activated protein kinase and protein phosphatase. Of interest, diclofenac had no effect on Na(+)-K(+) ATPases and Na(+)-K(+)-Cl- cotransporters, but inhibited cAMP-activated basolateral K(+) channels with IC50 of ∼ 3 µM. In addition, diclofenac suppressed Ca(2+)-activated Cl- channels, inwardly rectifying Cl- channels, and Ca(2+)-activated basolateral K(+) channels. Furthermore, diclofenac (up to 200 µM; 24 h of treatment) had no effect on cell viability and barrier function in T84 cells. Importantly, cholera toxin (CT)-induced Cl- secretion across T84 cell monolayers was effectively suppressed by diclofenac. Intraperitoneal administration of diclofenac (30 mg/kg) reduced both CT and Vibrio cholerae-induced intestinal fluid secretion by ∼ 70% without affecting intestinal fluid absorption in mice. Collectively, our results indicate that diclofenac inhibits both cAMP-activated and Ca(2+)-activated Cl- secretion by inhibiting both apical Cl- channels and basolateral K+ channels in intestinal epithelial cells. Diclofenac may be useful in the treatment of cholera and other types of secretory diarrheas resulting from intestinal

  2. Inhibition of cAMP-activated intestinal chloride secretion by diclofenac: cellular mechanism and potential application in cholera.

    Directory of Open Access Journals (Sweden)

    Pawin Pongkorpsakol

    2014-09-01

    Full Text Available Cyclic AMP-activated intestinal Cl- secretion plays an important role in pathogenesis of cholera. This study aimed to investigate the effect of diclofenac on cAMP-activated Cl- secretion, its underlying mechanisms, and possible application in the treatment of cholera. Diclofenac inhibited cAMP-activated Cl- secretion in human intestinal epithelial (T84 cells with IC50 of ∼ 20 µM. The effect required no cytochrome P450 enzyme-mediated metabolic activation. Interestingly, exposures of T84 cell monolayers to diclofenac, either in apical or basolateral solutions, produced similar degree of inhibitions. Analyses of the apical Cl- current showed that diclofenac reversibly inhibited CFTR Cl- channel activity (IC50 ∼ 10 µM via mechanisms not involving either changes in intracellular cAMP levels or CFTR channel inactivation by AMP-activated protein kinase and protein phosphatase. Of interest, diclofenac had no effect on Na(+-K(+ ATPases and Na(+-K(+-Cl- cotransporters, but inhibited cAMP-activated basolateral K(+ channels with IC50 of ∼ 3 µM. In addition, diclofenac suppressed Ca(2+-activated Cl- channels, inwardly rectifying Cl- channels, and Ca(2+-activated basolateral K(+ channels. Furthermore, diclofenac (up to 200 µM; 24 h of treatment had no effect on cell viability and barrier function in T84 cells. Importantly, cholera toxin (CT-induced Cl- secretion across T84 cell monolayers was effectively suppressed by diclofenac. Intraperitoneal administration of diclofenac (30 mg/kg reduced both CT and Vibrio cholerae-induced intestinal fluid secretion by ∼ 70% without affecting intestinal fluid absorption in mice. Collectively, our results indicate that diclofenac inhibits both cAMP-activated and Ca(2+-activated Cl- secretion by inhibiting both apical Cl- channels and basolateral K+ channels in intestinal epithelial cells. Diclofenac may be useful in the treatment of cholera and other types of secretory diarrheas resulting from intestinal

  3. 9 CFR 311.3 - Hog cholera.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Hog cholera. 311.3 Section 311.3... CERTIFICATION DISPOSAL OF DISEASED OR OTHERWISE ADULTERATED CARCASSES AND PARTS § 311.3 Hog cholera. (a) The carcasses of all hogs affected with hog cholera shall be condemned. (b) Inconclusive but suspicious symptoms...

  4. Global climate and infectious disease: the cholera paradigm.

    Science.gov (United States)

    Colwell, R R

    1996-12-20

    The origin of cholera has been elusive, even though scientific evidence clearly shows it is a waterborne disease. However, standard bacteriological procedures for isolation of the cholera vibrio from environmental samples, including water, between epidemics generally were unsuccessful. Vibrio cholerae, a marine vibrio, requiring salt for growth, enters into a dormant, viable but nonculturable stage when conditions are unfavorable for growth and reproduction. The association of Vibrio cholerae with plankton, notably copepods, provides further evidence for the environmental origin of cholera, as well as an explanation for the sporadic and erratic occurrence of cholera epidemics. On a global scale, cholera epidemics can now be related to climate and climatic events, such as El Niño, as well as the global distribution of the plankton host. Remote sensing, with the use of satellite imagery, offers the potential for predicting conditions conducive to cholera outbreaks or epidemics.

  5. Comparative Genomics of Vibrio cholerae O1 Isolated from Cholera Patients in Bangladesh

    DEFF Research Database (Denmark)

    Hossain, Zenat Zebin; Leekitcharoenphon, Pimlapas; Dalsgaard, Anders

    patients was co-infected with two V. cholerae strains (VC-1 and VC-3). Major virulence factors, biotype and antimicrobial resistance genes were identified by WGS. A global phylogenetic tree was inferred using genome wide SNPs (Single Nucleotide Polymorphism) analysis. RESULTS: All the V. cholerae strains...

  6. Cholera in Pregnancy: Outcomes from a Specialized Cholera Treatment Unit for Pregnant Women in L?og?ne, Haiti

    OpenAIRE

    Ciglenecki, Iza; Bichet, Mathieu; Tena, Javier; Mondesir, Erneau; Bastard, Mathieu; Tran, Nguyen-Toan; Antierens, Annick; Staderini, Nelly

    2013-01-01

    BACKGROUND: The association between cholera in pregnancy and negative fetal outcome has been described since the 19(th) century. However, there is limited published literature on the subject. We describe pregnancy outcomes from a specialized multidisciplinary hospital unit at the onset of a large cholera outbreak in Haiti in 2010 and 2011. METHODS: Pregnant women with cholera were hospitalized in a specialized unit within the MSF hospital compound in Léogâne and treated using standard cholera...

  7. Follicle-stimulating hormone receptor-mediated uptake of sup 45 Ca sup 2+ by cultured rat Sertoli cells does not require activation of cholera toxin- or pertussis toxin-sensitive guanine nucleotide binding proteins or adenylate cyclase

    Energy Technology Data Exchange (ETDEWEB)

    Grasso, P.; Reichert, L.E. Jr. (Albany Medical College, NY (USA))

    1990-08-01

    We have previously reported that FSH stimulates flux of 45Ca2+ into cultured Sertoli cells from immature rats via voltage-sensitive and voltage-independent calcium channels. In the present study, we show that this effect of FSH does not require cholera toxin (CT)- or pertussis toxin (PT)-sensitive guanine nucleotide binding (G) protein or activation of adenylate cyclase (AC). Significant stimulation of 45Ca2+ influx was observed within 1 min, and maximal response (3.2-fold over basal levels) was achieved within 2 min after exposure to FSH. FSH-stimulated elevations in cellular cAMP paralleled increases in 45Ca2+ uptake, suggesting a possible coupling of AC activation to 45Ca2+ influx. (Bu)2cAMP, however, was not able to enhance 45Ca2+ uptake over basal levels at a final concentration of 1000 microM, although a concentration-related increase in androstenedione conversion to estradiol was evident. Exposure of Sertoli cells to CT (10 ng/ml) consistently stimulated basal levels of androstenedione conversion to estradiol but had no effect on basal levels of 45Ca2+ uptake. Similarly, CT had no effect on FSH-induced 45Ca2+ uptake, but potentiated FSH-stimulated estradiol synthesis. PT (10 ng/ml) augmented basal and FSH-stimulated estradiol secretion without affecting 45Ca2+ influx. The adenosine analog N6-phenylisopropyladenosine, which binds to Gi-coupled adenosine receptors on Sertoli cells, inhibited FSH-stimulated androgen conversion to estradiol in a dose-related (1-1000 nM) manner, but FSH-stimulated 45Ca2+ influx remained unchanged. Our results show that in contrast to FSH-stimulated estradiol synthesis, the flux of 45Ca2+ into Sertoli cells in response to FSH is not mediated either directly or indirectly by CT- or PT-sensitive G protein, nor does it require activation of AC. Our data further suggest that the FSH receptor itself may function as a calcium channel.

  8. Afferent projections to the hamster intergeniculate leaflet demonstrated by retrograde and anterograde tracing

    DEFF Research Database (Denmark)

    Vrang, Niels; Mrosovsky, N.; Mikkelsen, Jens D.

    2003-01-01

    Circadian rhythms, Suprachiasmatic nucleus, Cholera toxin B, Phaseolus vulgaris-leucoagglutinin, Nonphotic......Circadian rhythms, Suprachiasmatic nucleus, Cholera toxin B, Phaseolus vulgaris-leucoagglutinin, Nonphotic...

  9. Epidemic Risk from Cholera Introductions into Mexico

    OpenAIRE

    Moore, Sean M.; Shannon, Kerry L.; Zelaya, Carla E.; Azman, Andrew S.; Lessler, Justin

    2014-01-01

    Stemming from the 2010 cholera outbreak in Haiti, cholera transmission in Hispaniola continues with over 40,000 cases in 2013. The presence of an ongoing cholera outbreak in the region poses substantial risks to countries throughout the Americas, particularly in areas with poor infrastructure. Since September 9, 2013 nearly 200 cholera cases have been reported in Mexico, as a result of introductions from Hispaniola or Cuba. There appear to have been multiple introductions into Mexico resultin...

  10. A Comparative Analysis of Vibrio cholerae Contamination in Point-of-Drinking and Source Water in a Low-Income Urban Community, Bangladesh.

    Science.gov (United States)

    Ferdous, Jannatul; Sultana, Rebeca; Rashid, Ridwan B; Tasnimuzzaman, Md; Nordland, Andreas; Begum, Anowara; Jensen, Peter K M

    2018-01-01

    Bangladesh is a cholera endemic country with a population at high risk of cholera. Toxigenic and non-toxigenic Vibrio cholerae ( V. cholerae ) can cause cholera and cholera-like diarrheal illness and outbreaks. Drinking water is one of the primary routes of cholera transmission in Bangladesh. The aim of this study was to conduct a comparative assessment of the presence of V. cholerae between point-of-drinking water and source water, and to investigate the variability of virulence profile using molecular methods of a densely populated low-income settlement of Dhaka, Bangladesh. Water samples were collected and tested for V. cholerae from "point-of-drinking" and "source" in 477 study households in routine visits at 6 week intervals over a period of 14 months. We studied the virulence profiles of V. cholerae positive water samples using 22 different virulence gene markers present in toxigenic O1/O139 and non-O1/O139 V. cholerae using polymerase chain reaction (PCR). A total of 1,463 water samples were collected, with 1,082 samples from point-of-drinking water in 388 households and 381 samples from 66 water sources. V. cholerae was detected in 10% of point-of-drinking water samples and in 9% of source water samples. Twenty-three percent of households and 38% of the sources were positive for V. cholerae in at least one visit. Samples collected from point-of-drinking and linked sources in a 7 day interval showed significantly higher odds ( P source [OR = 17.24 (95% CI = 7.14-42.89)] water. Based on the 7 day interval data, 53% (17/32) of source water samples were negative for V. cholerae while linked point-of-drinking water samples were positive. There were significantly higher odds ( p source water samples than in point-of-drinking water samples. Contamination of water at the point-of-drinking is less likely to depend on the contamination at the water source. Hygiene education interventions and programs should focus and emphasize on water at the point

  11. Updated Global Burden of Cholera in Endemic Countries

    Science.gov (United States)

    Ali, Mohammad; Nelson, Allyson R.; Lopez, Anna Lena; Sack, David A.

    2015-01-01

    Background The global burden of cholera is largely unknown because the majority of cases are not reported. The low reporting can be attributed to limited capacity of epidemiological surveillance and laboratories, as well as social, political, and economic disincentives for reporting. We previously estimated 2.8 million cases and 91,000 deaths annually due to cholera in 51 endemic countries. A major limitation in our previous estimate was that the endemic and non-endemic countries were defined based on the countries’ reported cholera cases. We overcame the limitation with the use of a spatial modelling technique in defining endemic countries, and accordingly updated the estimates of the global burden of cholera. Methods/Principal Findings Countries were classified as cholera endemic, cholera non-endemic, or cholera-free based on whether a spatial regression model predicted an incidence rate over a certain threshold in at least three of five years (2008-2012). The at-risk populations were calculated for each country based on the percent of the country without sustainable access to improved sanitation facilities. Incidence rates from population-based published studies were used to calculate the estimated annual number of cases in endemic countries. The number of annual cholera deaths was calculated using inverse variance-weighted average case-fatality rate (CFRs) from literature-based CFR estimates. We found that approximately 1.3 billion people are at risk for cholera in endemic countries. An estimated 2.86 million cholera cases (uncertainty range: 1.3m-4.0m) occur annually in endemic countries. Among these cases, there are an estimated 95,000 deaths (uncertainty range: 21,000-143,000). Conclusion/Significance The global burden of cholera remains high. Sub-Saharan Africa accounts for the majority of this burden. Our findings can inform programmatic decision-making for cholera control. PMID:26043000

  12. The Burden of Cholera in Uganda

    Science.gov (United States)

    Bwire, Godfrey; Malimbo, Mugagga; Maskery, Brian; Kim, Young Eun; Mogasale, Vittal; Levin, Ann

    2013-01-01

    Introduction In 2010, the World Health Organization released a new cholera vaccine position paper, which recommended the use of cholera vaccines in high-risk endemic areas. However, there is a paucity of data on the burden of cholera in endemic countries. This article reviewed available cholera surveillance data from Uganda and assessed the sufficiency of these data to inform country-specific strategies for cholera vaccination. Methods The Uganda Ministry of Health conducts cholera surveillance to guide cholera outbreak control activities. This includes reporting the number of cases based on a standardized clinical definition plus systematic laboratory testing of stool samples from suspected cases at the outset and conclusion of outbreaks. This retrospective study analyzes available data by district and by age to estimate incidence rates. Since surveillance activities focus on more severe hospitalized cases and deaths, a sensitivity analysis was conducted to estimate the number of non-severe cases and unrecognized deaths that may not have been captured. Results Cholera affected all ages, but the geographic distribution of the disease was very heterogeneous in Uganda. We estimated that an average of about 11,000 cholera cases occurred in Uganda each year, which led to approximately 61–182 deaths. The majority of these cases (81%) occurred in a relatively small number of districts comprising just 24% of Uganda's total population. These districts included rural areas bordering the Democratic Republic of Congo, South Sudan, and Kenya as well as the slums of Kampala city. When outbreaks occurred, the average duration was about 15 weeks with a range of 4–44 weeks. Discussion There is a clear subdivision between high-risk and low-risk districts in Uganda. Vaccination efforts should be focused on the high-risk population. However, enhanced or sentinel surveillance activities should be undertaken to better quantify the endemic disease burden and high-risk populations

  13. Epidemic risk from cholera introductions into Mexico.

    Science.gov (United States)

    Moore, Sean M; Shannon, Kerry L; Zelaya, Carla E; Azman, Andrew S; Lessler, Justin

    2014-02-21

    Stemming from the 2010 cholera outbreak in Haiti, cholera transmission in Hispaniola continues with over 40,000 cases in 2013. The presence of an ongoing cholera outbreak in the region poses substantial risks to countries throughout the Americas, particularly in areas with poor infrastructure. Since September 9, 2013 nearly 200 cholera cases have been reported in Mexico, as a result of introductions from Hispaniola or Cuba. There appear to have been multiple introductions into Mexico resulting in outbreaks of 2 to over 150 people. Using publicly available data, we attempt to estimate the reproductive number (R) of cholera in Mexico, and thereby assess the potential of continued introductions to establish a sustained epidemic. We estimate R for cholera in Mexico to be between 0.8 to 1.1, depending on the number of introductions, with the confidence intervals for the most plausible estimates crossing 1. These results suggest that the efficiency of cholera transmission in some regions of Mexico is near that necessary for a large epidemic. Intensive surveillance, evaluation of water and sanitation infrastructure, and planning for rapid response are warranted steps to avoid potential large epidemics in the region.

  14. Cholera toxin subunit B peptide fusion proteins reveal impaired oral tolerance induction in diabetes-prone but not in diabetes-resistant mice.

    Science.gov (United States)

    Presa, Maximiliano; Ortiz, Angela Zarama; Garabatos, Nahir; Izquierdo, Cristina; Rivas, Elisa I; Teyton, Luc; Mora, Conchi; Serreze, David; Stratmann, Thomas

    2013-11-01

    The cholera toxin B subunit (CTB) has been used as adjuvant to improve oral vaccine delivery in type 1 diabetes. The effect of CTB/peptide formulations on Ag-specific CD4(+) T cells has remained largely unexplored. Here, using tetramer analysis, we investigated how oral delivery of CTB fused to two CD4(+) T-cell epitopes, the BDC-2.5 T-cell 2.5 mi mimotope and glutamic acid decarboxylase (GAD) 286-300, affected diabetogenic CD4(+) T cells in nonobese diabetic (NOD) mice. When administered i.p., CTB-2.5 mi activated 2.5 mi(+) T cells and following intragastric delivery generated Ag-specific Foxp3(+) Treg and Th2 cells. While 2.5 mi(+) and GAD-specific T cells were tolerized in diabetes-resistant NODxB6.Foxp3(EGFP) F1 and nonobese resistant (NOR) mice, this did not occur in NOD mice. This indicated that NOD mice had a recessive genetic resistance to induce oral tolerance to both CTB-fused epitopes. In contrast to NODxB6.Foxp3(EGFP) F1 mice, oral treatment in NOD mice lead to strong 2.5 mi(+) T-cell activation and the sequestration of these cells to the effector-memory pool. Oral treatment of NOD mice with CTB-2.5 mi failed to prevent diabetes. These findings underline the importance of investigating the effect of oral vaccine formulations on diabetogenic T cells as in selected cases they may have counterproductive consequences in human patients. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. "Cystic fibrotics could survive cholera, choleraics could survive cystic fibrosis"; hypothesis that explores new horizons in treatment of cystic fibrosis.

    Science.gov (United States)

    Azimi, Arsalan

    2015-12-01

    Cystic fibrosis, the most common inherited disease of white population, is a disease of CFTR channels, in which mucosal function of many organs especially respiratory tract is impaired. Decreased mucociliary clearance and accumulation of mucus in airways facilitates colonization of infectious microorganisms, followed by infection. Following chronic infection, persistent inflammation ensues, which results in airway remodeling and deterioration of mucociliary clearance and result in a vicious cycle. Here, it is hypothesized that cholera toxin (CT) could ameliorate symptoms of cystic fibrosis as CT could dilute the thickened mucus, improve mucociliary clearance and alleviate airway obstruction. CT strengthens immunity of airway mucosa and it could attenuates bacterial growth and reduce persistency of infection. CT also modulates cellular immune response and it could decrease airway inflammation, hinder airway remodeling and prevent respiratory deterioration. Thereby it is hypothesized that CT could target and ameliorate many of pathophysiologic steps of the disease and it explores new horizons in treatment of CF. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Costs of Illness Due to Endemic Cholera

    Science.gov (United States)

    Poulos, C.; Riewpaiboon, A.; Stewart, J.F.; Clemens, J.; Guh, S.; Agtini, M.; Sur, D.; Islam, Z.; Lucas, M.; Whittington, D.

    2013-01-01

    Summary Economic analyses of cholera immunization programmes require estimates of the costs of cholera. The Diseases of the Most Impoverished programme measured the public, provider, and patient costs of culture-confirmed cholera in four study sites with endemic cholera using a combination of hospital- and community-based studies. Families with culture-proven cases were surveyed at home 7 and 14 days after confirmation of illness. Public costs were measured at local health facilities using a micro-costing methodology. Hospital-based studies found that the costs of severe cholera were USD 32 and 47 in Matlab and Beira. Community-based studies in North Jakarta and Kolkata found that cholera cases cost between USD 28 and USD 206, depending on hospitalization. Patient costs of illness as a percentage of average monthly income were 21% and 65% for hospitalized cases in Kolkata and North Jakarta, respectively. This burden on families is not captured by studies that adopt a provider perspective. PMID:21554781

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

    Directory of Open Access Journals (Sweden)

    Pappu Kumar Gupta

    2016-06-01

    Full Text Available Abstract 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 from watery diarrhea during July to December 2012 were obtained from outbreak areas from all over Nepal. Alkaline peptone water and thiosulphate citrate bile salt sucrose agar (TCBS were used to isolate the Vibrio cholerae. The isolates were identified with the help of colony morphology, Gram’s staining, conventional biochemical testing, serotyping and biotyping. Antimicrobial susceptibility testing was performed by determining the minimum inhibitory concentration (MIC by agar dilution method. Results Vibrio cholerae was isolated from 26.72 % of total samples. All isolated Vibrio cholerae were confirmed to be Vibrio cholerae serogoup O1 biotype El Tor and serotype Ogawa. All isolates were resistant to ampicillin and cotrimoxazole. Twenty nine isolates were resistant toward two different classes of antibiotics, one strain was resistant to three different classes of antibiotics and one strain was resistant to four different classes of antibiotics. According to the definition of the multidrug resistant bacteria; 6.45 % of the strains of Vibrio cholerae were found to be multidrug resistant. Conclusions Cholera due to multidrug resistant Vibrio cholerae is also possible in Nepal. According to the antimicrobial susceptibility pattern of Vibrio cholerae in our study we recommend to use any antibiotics among tetracycline, doxycycline, levofloxacin, azithromycin, chloramphenicol and ciprofloxacin for preliminary treatment of cholera in Nepal.

  18. Cholera ante portas – The re-emergence of cholera in Kinshasa after a ten-year hiatus

    Science.gov (United States)

    Bompangue, Didier; Vesenbeckh, Silvan Manuel; Giraudoux, Patrick; Castro, Marcia; Muyembe, Jean-Jacques; Kebela Ilunga, Benoît; Murray, Megan

    2012-01-01

    Background: Cholera is an endemic disease in certain well-defined areas in the east of the Democratic Republic of Congo (DRC). The west of the country, including the mega-city Kinshasa, has been free of cases since mid 2001 when the last outbreak ended. Methods and Findings: We used routinely collected passive surveillance data to construct epidemic curves of the cholera cases and map the spatio-temporal progress of the disease during the first 47 weeks of 2011. We compared the spatial distribution of disease spread to that which occurred in the last cholera epidemic in Kinshasa between 1996 and 2001. To better understand previous determinants of cholera spread in this region, we conducted a correlation analysis to assess the impact of rainfall on weekly health zone cholera case counts between December 1998 and March 2001 and a Generalized Linear Model (GLM) regression analysis to identify factors that have been associated with the most vulnerable health zones within Kinshasa between October 1998 and June 1999. In February 2011, cholera reemerged in a region surrounding Kisangani and gradually spread westwards following the course of the Congo River to Kinshasa, home to 10 million people. Ten sampled isolates were confirmed to be Vibrio cholerae O1, biotype El Tor, serotype Inaba, resistant to trimethoprim-sulfa, furazolidone, nalidixic acid, sulfisoxaole, and streptomycin, and intermediate resistant to Chloramphenicol. An analysis of a previous outbreak in Kinshasa shows that rainfall was correlated with case counts and that health zone population densities as well as fishing and trade activities were predictors of case counts. Conclusion: Cholera is particularly difficult to tackle in the DRC. Given the duration of the rainy season and increased riverine traffic from the eastern provinces in late 2011, we expect further increases in cholera in the coming months and especially within the mega-city Kinshasa. We urge all partners involved in the response to remain

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

    OpenAIRE

    Yan Boucher

    2016-01-01

    ABSTRACT 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 coun...

  20. Use of a real time PCR assay for detection of the ctxA gene of Vibrio cholerae in an environmental survey of Mobile Bay.

    Science.gov (United States)

    Blackstone, George M; Nordstrom, Jessica L; Bowen, Michael D; Meyer, Richard F; Imbro, Paula; DePaola, Angelo

    2007-02-01

    Toxigenic Vibrio cholerae, the etiological agent of cholera, is a natural inhabitant of the marine environment and causes severe diarrheal disease affecting thousands of people each year in developing countries. It is the subject of extensive testing of shrimp produced and exported from these countries. We report the development of a real time PCR (qPCR) assay to detect the gene encoding cholera toxin, ctxA, found in toxigenic V. cholerae strains. This assay was tested against DNA isolated from soil samples collected from diverse locations in the US, a panel of eukaryotic DNA from various sources, and prokaryotic DNA from closely related and unrelated bacterial sources. Only Vibrio strains known to contain ctxA generated a fluorescent signal with the 5' nuclease probe targeting the ctxA gene, thus confirming the specificity of the assay. In addition, the assay was quantitative in pure culture across a six-log dynamic range down to <10 CFU per reaction. To test the robustness of this assay, oysters, aquatic sediments, and seawaters from Mobile Bay, AL, were analyzed by qPCR and traditional culture methods. The assay was applied to overnight alkaline peptone water enrichments of these matrices after boiling the enrichments for 10 min. Toxigenic V. cholerae strains were not detected by either qPCR or conventional methods in the 16 environmental samples examined. A novel exogenous internal amplification control developed by us to prevent false negatives identified the samples that were inhibitory to the PCR. This assay, with the incorporated internal control, provides a highly specific, sensitive, and rapid detection method for the detection of toxigenic strains of V. cholerae.

  1. Human Gut Microbiota Predicts Susceptibility to Vibrio cholerae Infection.

    Science.gov (United States)

    Midani, Firas S; Weil, Ana A; Chowdhury, Fahima; Begum, Yasmin A; Khan, Ashraful I; Debela, Meti D; Durand, Heather K; Reese, Aspen T; Nimmagadda, Sai N; Silverman, Justin D; Ellis, Crystal N; Ryan, Edward T; Calderwood, Stephen B; Harris, Jason B; Qadri, Firdausi; David, Lawrence A; LaRocque, Regina C

    2018-04-12

    Cholera is a public health problem worldwide and the risk factors for infection are only partially understood. We prospectively studied household contacts of cholera patients to compare those who were infected with those who were not. We constructed predictive machine learning models of susceptibility using baseline gut microbiota data. We identified bacterial taxa associated with susceptibility to Vibrio cholerae infection and tested these taxa for interactions with V. cholerae in vitro. We found that machine learning models based on gut microbiota predicted V. cholerae infection as well as models based on known clinical and epidemiological risk factors. A 'predictive gut microbiota' of roughly 100 bacterial taxa discriminated between contacts who developed infection and those who did not. Susceptibility to cholera was associated with depleted levels of microbes from the phylum Bacteroidetes. By contrast, a microbe associated with cholera by our modeling framework, Paracoccus aminovorans, promoted the in vitro growth of V. cholerae. Gut microbiota structure, clinical outcome, and age were also linked. These findings support the hypothesis that abnormal gut microbial communities are a host factor related to V. cholerae susceptibility.

  2. Effectiveness of an oral cholera vaccine campaign to prevent clinically-significant cholera in Odisha State, India.

    Science.gov (United States)

    Wierzba, Thomas F; Kar, Shantanu K; Mogasale, Vijayalaxmi V; Kerketta, Anna S; You, Young Ae; Baral, Prameela; Khuntia, Hemant K; Ali, Mohammad; Kim, Yang Hee; Rath, Shyam Bandhu; Bhattachan, Anuj; Sah, Binod

    2015-05-15

    A clinical trial conducted in India suggests that the oral cholera vaccine, Shanchol, provides 65% protection over five years against clinically-significant cholera. Although the vaccine is efficacious when tested in an experimental setting, policymakers are more likely to use this vaccine after receiving evidence demonstrating protection when delivered to communities using local health department staff, cold chain equipment, and logistics. We used a test-negative, case-control design to evaluate the effectiveness of a vaccination campaign using Shanchol and validated the results using a cohort approach that addressed disparities in healthcare seeking behavior. The campaign was conducted by the local health department using existing resources in a cholera-endemic area of Puri District, Odisha State, India. All non-pregnant residents one year of age and older were offered vaccine. Over the next two years, residents seeking care for diarrhea at one of five health facilities were asked to enroll following informed consent. Cases were patients seeking treatment for laboratory-confirmed V. cholera-associated diarrhea. Controls were patients seeking treatment for V. cholerae negative diarrhea. Of 51,488 eligible residents, 31,552 individuals received one dose and 23,751 residents received two vaccine doses. We identified 44 V. cholerae O1-associated cases and 366 non V. cholerae diarrhea controls. The adjusted protective effectiveness for persons receiving two doses was 69.0% (95% CI: 14.5% to 88.8%), which is similar to the adjusted estimates obtained from the cohort approach. A statistical trend test suggested a single dose provided a modicum of protection (33%, test for trend, p=0.0091). This vaccine was found to be as efficacious as the results reported from a clinical trial when administered to a rural population using local health personnel and resources. This study provides evidence that this vaccine should be widely deployed by public health departments in

  3. 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

  4. Cholera: an overview with reference to the Yemen epidemic.

    Science.gov (United States)

    Rabaan, Ali A

    2018-06-22

    Cholera is a secretory diarrhoeal disease caused by infection with Vibrio cholerae, primarily the V. cholerae O1 El Tor biotype. There are approximately 2.9 million cases in 69 endemic countries annually, resulting in 95 000 deaths. Cholera is associated with poor infrastructure and lack of access to sanitation and clean drinking water. The current cholera epidemic in Yemen, linked to spread of V. cholerae O1 (Ogawa serotype), is associated with the ongoing war. This has devastated infrastructure and health services. The World Health Organization had estimated that 172 286 suspected cases arose between 27th April and 19th June 2017, including 1170 deaths. While there are three oral cholera vaccines prequalified by the World Health Organization, there are issues surrounding vaccination campaigns in conflict situations, exacerbated by external factors such as a global vaccine shortage. Major movements of people complicates surveillance and administration of double doses of vaccines. Cholera therapy mainly depends on rehydration, with use of antibiotics in more severe infections. Concerns have arisen about the rise of antibiotic resistance in cholera, due to mobile genetic elements. In this review, we give an overview of cholera epidemiology, virulence, antibiotic resistance, therapy and vaccines, in the light of the ongoing epidemic in Yemen.

  5. Efficacy of Ciprofloxacin for Treatment of Cholera Associated with Diminished Susceptibility to Ciprofloxacin to Vibrio cholerae O1.

    Science.gov (United States)

    Khan, Wasif Ali; Saha, Debasish; Ahmed, Sabeena; Salam, Mohammed Abdus; Bennish, Michael Louis

    2015-01-01

    We identified a poor clinical response to treatment of cholera with a single 1 g dose of ciprofloxacin, a standard treatment for cholera. To determine reasons for the poor response and better therapeutic approaches we examined the minimal inhibitor concentration (MIC, n = 275) and disc-diffusion zone sizes (n = 205) for ciprofloxacin and nalidixic acid of V. cholerae O1 strains isolated in Bangladesh from 1994 to 2012, and reexamined data from 161 patients infected with Vibrio cholerae O1 recruited in four clinical trials who received single- or multiple-dose ciprofloxacin for treatment of cholera and compared their clinical response to the V. cholerae O1 susceptibility. Although all 275 isolates of V. cholerae O1 remained susceptible to ciprofloxacin using standard MIC and disc-diffusion thresholds, the MIC90 to ciprofloxacin increased from 0.010 in 1994 to 0.475 μgm/ml in 2012. Isolates became frankly resistant to nalidixic with the MIC90 increasing from 21 μgm/ml in 1994 to >256 μgm/ml and 166 of 205 isolates from 1994 to 2005 being frankly resistant using disc-diffusion testing. Isolates resistant to nalidixic acid by disc-diffusion testing had a median ciprofloxacin MIC of 0.190 μgm/ml (10th-90th centiles 0.022 to 0.380); nalidixic acid-susceptible isolates had a median ciprofloxacin MIC of 0.002 (0.002 to 0.012).The rate of clinical success with single-dose ciprofloxacin treatment for nalidixic acid-susceptible strains was 94% (61 of 65 patients) and bacteriologic success 97% (63/65) compared to 18% (12/67) and 8% (5/67) respectively with nalidixic acid-resistant strains (Ptreatment with ciprofloxacin had 86% and 100% clinical and bacteriologic success rates respectively in patients infected with nalidixic acid-susceptible strains of V. cholerae O1 compared to clinical success 67% and bacteriologic success 60% with nalidixic acid-resistant strains. Single-dose ciprofloxacin is not effective for treating cholera caused by V. cholerae O1 with diminished

  6. [The cholera epidemic in Latin America].

    Science.gov (United States)

    Olsvik, O

    1992-05-30

    An outbreak of cholera started in Peru in January 1991 and spread through most Latin American countries within a year. This was the first known epidemic of cholera in America for more than a century. In 1991, 321,334 persons were reported to have cholera in Peru, 119,063 were hospitalized, and 2,906 died. Other countries like Ecuador, Colombia, Guatemala, Brazil, Mexico, Bolivia, Chile, El Salvador, Venezuela and Honduras were also affected, but these countries combined accounted for only 20% of the cases registered in Peru. In April 1992, all Latin American countries except Uruguay, Paraguay and French Guyana have reported cholera. The mortality rate for the epidemic in Latin America was only 1%, mainly owing to good oral rehydration treatment provided by Local health services and the Pan American Health Organization. The causative organism was Vibrio cholerae, serogroup O1, serotype Inaba (and Ogawa) of the El Tor biotype. Genetic characterization shows this strain to be unique, and the designation is reserved for the Latin American strain, distinguishing it from the other El Tor isolates from the 7th pandemic.

  7. Cellular vacuoles induced by Mycoplasma pneumoniae CARDS toxin originate from Rab9-associated compartments.

    Directory of Open Access Journals (Sweden)

    Coreen Johnson

    Full Text Available Recently, we identified an ADP-ribosylating and vacuolating cytotoxin in Mycoplasma pneumoniae designated Community Acquired Respiratory Distress Syndrome (CARDS toxin. In this study we show that vacuoles induced by recombinant CARDS (rCARDS toxin are acidic and derive from the endocytic pathway as determined by the uptake of neutral red and the fluid-phase marker, Lucifer yellow, respectively. Also, we demonstrate that the formation of rCARDS toxin-associated cytoplasmic vacuoles is inhibited by the vacuolar ATPase inhibitor, bafilomycin A1, and the ionophore, monensin. To examine the ontogeny of these vacuoles, we analyzed the distribution of endosomal and lysosomal membrane markers during vacuole formation and observed the enrichment of the late endosomal GTPase, Rab9, around rCARDS toxin-induced vacuoles. Immunogold-labeled Rab9 and overexpression of green fluorescent-tagged Rab9 further confirmed vacuolar association. The late endosomal- and lysosomal-associated membrane proteins, LAMP1 and LAMP2, also localized to the vacuolar membranes, while the late endosomal protein, Rab7, and early endosomal markers, Rab5 and EEA1, were excluded. HeLa cells expressing dominant-negative (DN Rab9 exhibited markedly reduced vacuole formation in the presence of rCARDS toxin, in contrast to cells expressing DN-Rab7, highlighting the importance of Rab9 function in rCARDS toxin-induced vacuolation. Our findings reveal the unique Rab9-association with rCARDS toxin-induced vacuoles and its possible relationship to the characteristic histopathology that accompanies M. pneumoniae infection.

  8. Diversity and Impact of Prokaryotic Toxins on Aquatic Environments: A Review

    Directory of Open Access Journals (Sweden)

    Rogério Tenreiro

    2010-10-01

    Full Text Available Microorganisms are ubiquitous in all habitats and are recognized by their metabolic versatility and ability to produce many bioactive compounds, including toxins. Some of the most common toxins present in water are produced by several cyanobacterial species. As a result, their blooms create major threats to animal and human health, tourism, recreation and aquaculture. Quite a few cyanobacterial toxins have been described, including hepatotoxins, neurotoxins, cytotoxins and dermatotoxins. These toxins are secondary metabolites, presenting a vast diversity of structures and variants. Most of cyanobacterial secondary metabolites are peptides or have peptidic substructures and are assumed to be synthesized by non-ribosomal peptide synthesis (NRPS, involving peptide synthetases, or NRPS/PKS, involving peptide synthetases and polyketide synthases hybrid pathways. Besides cyanobacteria, other bacteria associated with aquatic environments are recognized as significant toxin producers, representing important issues in food safety, public health, and human and animal well being. Vibrio species are one of the most representative groups of aquatic toxin producers, commonly associated with seafood-born infections. Some enterotoxins and hemolysins have been identified as fundamental for V. cholerae and V. vulnificus pathogenesis, but there is evidence for the existence of other potential toxins. Campylobacter spp. and Escherichia coli are also water contaminants and are able to produce important toxins after infecting their hosts. Other bacteria associated with aquatic environments are emerging as toxin producers, namely Legionella pneumophila and Aeromonas hydrophila, described as responsible for the synthesis of several exotoxins, enterotoxins and cytotoxins. Furthermore, several Clostridium species can produce potent neurotoxins. Although not considered aquatic microorganisms, they are ubiquitous in the environment and can easily contaminate drinking

  9. Tracking micro-optical resonances for identifying and sensing novel procaspase-3 protein marker released from cell cultures in response to toxins

    International Nuclear Information System (INIS)

    Chen, Ying-Jen; Vollmer, Frank; Xiang, Wei; Klucken, Jochen

    2016-01-01

    The response of cells to toxins is commonly investigated by detecting intracellular markers for cell death, such as caspase proteins. This requires the introduction of labels by the permeabilization or complete lysis of cells. Here we introduce a non-invasive tool for monitoring a caspase protein in the extracellular medium. The tool is based on highly sensitive optical micro-devices, referred to as whispering-gallery mode biosensors (WGMBs). WGMBs are functionalized with antibodies for the specific and label-free detection of procaspase-3 released from human embryonic kidney HEK293 and neuroglioma H4 cells after introducing staurosporine and rotenone toxins, respectively. Additional tests show that the extracellular accumulation of procaspase-3 is concomitant with a decrease in cell viability. The hitherto unknown release of procaspase-3 from cells in response to toxins and its accumulation in the medium is further investigated by Western blot, showing that the extracellular detection of procaspase-3 is interrelated with cytotoxicity of alpha-synuclein protein (aSyn) overexpressed in H4 cells. These studies provide evidence for procaspase-3 as a novel extracellular biomarker for cell death, with applications in cytotoxicity tests. Such WGMBs could be applied to further identify as-yet unknown extracellular biomarkers using established antibodies against intracellular antigens. (paper)

  10. The health economics of cholera: A systematic review.

    Science.gov (United States)

    Hsiao, Amber; Hall, Angela H; Mogasale, Vittal; Quentin, Wilm

    2018-06-12

    Vibrio cholera is a major contributor of diarrheal illness that causes significant morbidity and mortality globally. While there is literature on the health economics of diarrheal illnesses more generally, few studies have quantified the cost-of-illness and cost-effectiveness of cholera-specific prevention and control interventions. The present systematic review provides a comprehensive overview of the literature specific to cholera as it pertains to key health economic measures. A systematic review was performed with no date restrictions up through February 2017 in PubMed, Econlit, Embase, Web of Science, and Cochrane Review to identify relevant health economics of cholera literature. After removing duplicates, a total of 1993 studies were screened and coded independently by two reviewers, resulting in 22 relevant studies. Data on population, methods, and results (cost-of-illness and cost-effectiveness of vaccination) were compared by country/region. All costs were adjusted to 2017 USD for comparability. Costs per cholera case were found to be rather low: $1000/case. There is adequate evidence to support the economic value of vaccination for the prevention and control of cholera when vaccination is targeted at high-incidence populations and/or areas with high case fatality rates due to cholera. When herd immunity is considered, vaccination also becomes a cost-effective option for the general population and is comparable in cost-effectiveness to other routine immunizations. Cholera vaccination is a viable short-to-medium term option, especially as the upfront costs of building water, sanitation, and hygiene (WASH) infrastructure are considerably higher for countries that face a significant burden of cholera. While WASH may be the more cost-effective solution in the long-term when implemented properly, cholera vaccination can still be a feasible, cost-effective strategy. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. A unique role of the cholera toxin A1-DD adjuvant for long-term plasma and memory B cell development.

    Science.gov (United States)

    Bemark, Mats; Bergqvist, Peter; Stensson, Anneli; Holmberg, Anna; Mattsson, Johan; Lycke, Nils Y

    2011-02-01

    Adjuvants have traditionally been appreciated for their immunoenhancing effects, whereas their impact on immunological memory has largely been neglected. In this paper, we have compared three mechanistically distinct adjuvants: aluminum salts (Alum), Ribi (monophosphoryl lipid A), and the cholera toxin A1 fusion protein CTA1-DD. Their influence on long-term memory development was dramatically different. Whereas a single immunization i.p. with 4-hydroxy-3-nitrophenyl acetyl (NP)-chicken γ-globulin and adjuvant stimulated serum anti-NP IgG titers that were comparable at 5 wk, CTA1-DD-adjuvanted responses were maintained for >16 mo with a half-life of anti-NP IgG ∼36 wk, but DD dose-dependent increase in germinal center (GC) size and numbers was found, with >60% of splenic B cell follicles hosting GC at an optimal CTA1-DD dose. Roughly 7% of these GC were NP specific. This GC-promoting effect correlated well with the persistence of long-term plasma cells in the bone marrow and memory B cells in the spleen. CTA1-DD also facilitated increased somatic hypermutation and affinity maturation of NP-specific IgG Abs in a dose-dependent fashion, hence arguing that large GC not only promotes higher Ab titers but also high-quality Ab production. Adoptive transfer of splenic CD80(+), but not CD80(-), B cells, at 1 y after immunization demonstrated functional long-term anti-NP IgG and IgM memory cells. To our knowledge, this is the first report to specifically compare and document that adjuvants can differ considerably in their support of long-term immune responses. Differential effects on the GC reaction appear to be the basis for these differences.

  12. Combating cholera epidemics by targeting reservoirs of infection ...

    African Journals Online (AJOL)

    Objectives: To determine the parameters which can be investigated for prevention and effective control of cholera. Data sources: Literature search on compact disk-read only memory (CD-ROM), medline and internet, using the key words: cholera outbreaks, and cholera transmission. A few reviews were manually reviewed.

  13. Avian cholera

    Science.gov (United States)

    Friend, Milton

    1999-01-01

    Avian cholera is a contagious disease resulting from infection by the bacterium Pasteurella multocida. Several subspecies of bacteria have been proposed for P. multocida, and at least 16 different P. multocida serotypes or characteristics of antigens in bacterial cells that differentiate bacterial variants from each other have been recognized. The serotypes are further differentiated by other methods, including DNA fingerprinting. These evaluations are useful for studying the ecology of avian cholera (Fig. 7.1), because different serotypes are generally found in poultry and free-ranging migratory birds. These evaluations also show that different P. multocida serotypes are found in wild birds in the eastern United States than those that are found in the birds in the rest of the Nation (Fig. 7.2).

  14. Knowledge of, attitudes toward, and preventive practices relating to cholera and oral cholera vaccine among urban high-risk groups: findings of a cross-sectional study in Dhaka, Bangladesh

    Science.gov (United States)

    2013-01-01

    Background In endemic countries such as Bangladesh, consequences of cholera place an enormous financial and social burden on patients and their families. Cholera vaccines not only provide health benefits to susceptible populations but also have effects on the earning capabilities and financial stability of the family. Community-based research and evaluations are necessary to understand perceptions about and practices of the community relating to cholera and oral cholera vaccines. This may help identify the ways in which such vaccines may be successfully introduced, and other preventive measures can be implemented. The present study assessed the knowledge of, attitudes toward, and preventive practices relating to cholera and oral cholera vaccine among an urban population residing in a high cholera-prone setting in Dhaka, Bangladesh. Methods This cross-sectional study was conducted in an area of high cholera prevalence in 15 randomly-selected clusters in Mirpur, Dhaka city. A study team collected data through a survey and in-depth interviews during December 2010–February 2011. Results Of 2,830 families included in the final analysis, 23% could recognize cholera as acute watery diarrhea and 16% had ever heard of oral cholera vaccine. About 54% of the respondents had poor knowledge about cholera-related issues while 97% had a positive attitude toward cholera and oral cholera vaccine. One-third showed poor practice relating to the prevention of cholera. The findings showed a significant (p cholera were the significant predictors to having poor knowledge. Conclusions The findings suggest the strengthening of health education activities to improve knowledge on cholera, its prevention and treatment and information on cholera vaccination among high-risk populations. The data also underscore the potential of mass cholera vaccination to prevent and control cholera. PMID:23509860

  15. 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)

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

    2016-01-01

    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. 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. 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.

  16. [Seroepidemiology of cholera in Mexico].

    Science.gov (United States)

    González-Bonilla, C; Valle-Valdez, J G; Núñez-León, A; Moguel-Pech, L; Villanueva-Zamudio, A

    1994-01-01

    Antibodies against Vibrio cholerae were determined in 2352 serum samples obtained from patients with clinical diagnosis of cholera. Samples from their contacts and from healthy people living in the same communities were also analyzed. Vibriocidal antibodies with titers 1:160 or higher were observed in 25% of the samples. An increase of vibriocidal and antitoxin antibody titers were observed in 56 to 60% of the patients in which paired samples were available, one obtained in the acute phase of the disease and the other in the convalescence, confirming the diagnosis of cholera. Differences in the antibody titers were noticed when comparing the serotype according to the geographic area and the season of the year.

  17. Projections from the raphe nuclei to the suprachiasmatic nucleus of the rat

    DEFF Research Database (Denmark)

    Hay-Schmidt, Anders; Vrang, N.; Larsen, P.J.

    2003-01-01

    Hypothalamus, Circadian rhythm, Serotonin, Nucleus, Neuronal connections, Phaseolus vulgaris-leucoagglutinin (PHA-L), Cholera toxin (ChB)......Hypothalamus, Circadian rhythm, Serotonin, Nucleus, Neuronal connections, Phaseolus vulgaris-leucoagglutinin (PHA-L), Cholera toxin (ChB)...

  18. Cholera in pregnancy: outcomes from a specialized cholera treatment unit for pregnant women in Léogâne, Haiti.

    Science.gov (United States)

    Ciglenecki, Iza; Bichet, Mathieu; Tena, Javier; Mondesir, Erneau; Bastard, Mathieu; Tran, Nguyen-Toan; Antierens, Annick; Staderini, Nelly

    2013-01-01

    The association between cholera in pregnancy and negative fetal outcome has been described since the 19(th) century. However, there is limited published literature on the subject. We describe pregnancy outcomes from a specialized multidisciplinary hospital unit at the onset of a large cholera outbreak in Haiti in 2010 and 2011. Pregnant women with cholera were hospitalized in a specialized unit within the MSF hospital compound in Léogâne and treated using standard cholera treatment guidelines but with earlier, more intense fluid replacement. All women had intravenous access established at admission regardless of their hydration status, and all received antibiotic treatment. Data were collected on patient demographics, pregnancy and cholera status, and pregnancy outcome. In this analysis we calculated risk ratios for fetal death and performed logistic regression analysis to control for confounding factors. 263 pregnant women with cholera were hospitalized between December 2010 and July 2011. None died during hospitalization, 226 (86%) were discharged with a preserved pregnancy and 16 (6%) had live fullterm singleton births, of whom 2 died within the first 5 days postpartum. The remaining 21 pregnancies (8%) resulted in intrauterine fetal death. The risk of fetal death was associated with factors reflecting severity of the cholera episode: after adjusting for confounding factors, the strongest risk factor for fetal death was severe maternal dehydration (adjusted risk ratio for severe vs. mild dehydration was 9.4, 95% CI 2.5-35.3, p = 0.005), followed by severe vomiting (adjusted risk ratio 5.1, 95% 1.1-23.8, p = 0.041). This is the largest cohort of pregnant women with cholera described to date. The main risk factor identified for fetal death was severity of dehydration. Our experience suggests that establishing specialized multidisciplinary units which facilitate close follow-up of both pregnancy and dehydration status due to cholera could be beneficial

  19. Two forms of Vibrio cholerae O1 El Tor hemolysin derived from identical precursor protein.

    Science.gov (United States)

    Ikigai, H; Ono, T; Nakae, T; Otsuru, H; Shimamura, T

    1999-01-08

    Vibrio cholerae O1 grown in heart infusion broth produces two forms of El Tor hemolysin (ETH) monomers of 65 and 50 kDa. These monomers form several different sizes of mixed oligomers ranging from 180 to 280 kDa in the liposomal membranes. We found that the N-terminal amino acid sequences, NH2-Trp-Pro-Ala-Pro-Ala-Asn-Ser-Glu, of both the 65- and 50-kDa toxins were identical. We assumed, therefore, that the 65- and 50-kDa toxins were derivatives of the identical precursor protein and the 50-kDa protein was a truncated derivative of 65-kDa ETH. To substantiate this assumption, we treated the 260-kDa oligomer with trypsin and obtained a 190-kDa oligomer. This 190-kDa oligomer consisted of only the 50-kDa subunits. Both 260- and 190-kDa oligomers formed ion channels indistinguishable from each other in planar lipid bilayers. These results suggest that the essential part of the ETH in forming the membrane-damaging aggregate is a 50-kDa protein.

  20. 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.

  1. Geographical patterns of cholera in Mexico, 1991-1996.

    Science.gov (United States)

    Borroto, R J; Martinez-Piedra, R

    2000-08-01

    The seventh cholera pandemic has been ongoing in Mexico since 1991 and threatens to become endemic. This paper aims to determine the geographical pattern of cholera in Mexico to define areas at high risk of endemic cholera. Ecologic research was conducted based upon the cartography of disease incidence. The 32 Mexican states were grouped into five strata according to the value of the 1991-1996 cumulative incidence rate of cholera. Rate ratios were computed for strata of states classified by geographical situation, urbanization, and poverty level. Cholera incidence was 2.47 times higher in coastal states than in the interior (95% CI : 2.42-2.52). The disease was negatively associated with urbanization. Incidence in the least urbanized stratum was four times as high as in the most urban stratum (95% CI : 3.9-4.12). The poorest stratum showed the most remarkable incidence, i.e. 5.9 times higher than the rate in the least poor stratum (95% CI : 5.73-6.04). This ecologic research suggests that high poverty level, low urbanization, and southern location are the most important predictors of endemic cholera in Mexican states. It is hypothesized that the natural environment of the coastal plains in southern states may also play a significant role in cholera incidence. Poor communities residing in the southern, predominantly rural, coastal states should be prioritized when it comes to investing in safe water supply facilities, adequate excreta disposal systems and cholera surveillance.

  2. Investigation of household contamination of Vibrio cholerae in Bangladesh

    DEFF Research Database (Denmark)

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

    . cholerae El Tor strain N16961, showed hemolysis and proteolysis activity but none of them exhibited any hemagglutinin activity on human erythrocytes. The study findings indicate that V. cholerae contamination is mostly originated in and around kitchen area rather than latrine area. Contaminated food...... 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......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...

  3. Cholera Epidemiology in Nigeria: an overview | Adagbada | Pan ...

    African Journals Online (AJOL)

    Cholera is an acute diarrhoeal infection caused by ingestion of food or water contaminated with the bacterium, Vibrio cholera. Choleragenic V. cholera O1 and O139 are the only causative agents of the disease. The two most distinguishing epidemiologic features of the disease are its tendency to appear in explosive ...

  4. Distribution ofVibrio cholerae in two Florida estuaries.

    Science.gov (United States)

    Hood, M A; Ness, G E; Rodrick, G E; Blake, N J

    1983-04-01

    The distribution ofVibrio cholerae was examined in 2 Florida estuaries, Apalachicola and Tampa Bay.Vibrio cholerae serotype non-01 was the most abundant serotype, being isolated from 45% of the oyster samples, 30% of the sediments, 50% of the waters, and 75% of the blue crabs.Vibrio cholerae serotype 01 was isolated from only one oyster sample. Strong linear correlations betweenV. cholerae and temperature, salinity, or the other physical/chemical parameters measured,Escherichia coli, or fecal coliforms were not observed, but a range of temperatures and salinities appeared relevant to the distribution of the organism. The organism was present in the highest concentrations when salinities were 10‰-25‰ and temperatures were 20‡C-35‡C.In vitro growth curves of 95V. cholerae environmental isolates further supported that 10‰-25‰ was an ideal salinity range for the organisms. The results suggest thatV. cholerae is a widely distributed organism in the nutrient-rich warm waters of the Gulf Coast estuaries.

  5. Killed oral cholera vaccines: history, development and implementation challenges.

    Science.gov (United States)

    Lopez, Anna Lena; Gonzales, Maria Liza Antoinette; Aldaba, Josephine G; Nair, G Balakrish

    2014-09-01

    Cholera is still a major global health problem, affecting mainly people living in unsanitary conditions and who are at risk for outbreaks of cholera. During the past decade, outbreaks are increasingly reported from more countries. From the early killed oral cholera vaccine, rapid improvements in vaccine development occurred as a result of a better understanding of the epidemiology of the disease, pathogenesis of cholera infection and immunity. The newer-generation oral killed cholera vaccines have been shown to be safe and effective in field trials conducted in cholera endemic areas. Likewise, they have been shown to be protective when used during outbreak settings. Aside from providing direct protection to vaccinated individuals, recent studies have demonstrated that these killed oral vaccines also confer indirect protection through herd immunity. Although new-generation oral cholera vaccines should not be considered in isolation from other preventive approaches in countries where they are most needed, especially improved water quality and sanitation, these vaccines serve as immediately available public health tools for preventing further morbidity and mortality from cholera. However, despite its availability for more than two decades, use of these vaccines has not been optimized. Although there are limitations of the currently available oral cholera vaccines, recent data show that the vaccines are safe, feasible to use even in difficult circumstances and able to provide protection in various settings. Clear identification of the areas and target population groups who will benefit from the use of the cholera vaccines will be required and strategies to facilitate accessibility and usage of these vaccines in these areas and population groups will need to be developed.

  6. High case fatality cholera outbreak in Western Kenya, August 2010

    African Journals Online (AJOL)

    abp

    Abstract. Introduction: Cholera is a disease caused by the bacterium Vibrio cholera and has been an important public health problem since its first pandemic in 1817. Kenya has had numerous outbreaks of cholera ever since it was first detected there during 1971. In August 2010 an outbreak of cholera occurred in Kuria ...

  7. Antimicrobial drugs for treating cholera

    Science.gov (United States)

    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 differences between classes of antimicrobials or dosing schedules. Search methods We searched the Cochrane Infectious Disease Group Specialized Register; the Cochrane Central Register of Controlled Trials (CENTRAL); PubMed; EMBASE; African Index Medicus; LILACS; Science Citation Index; metaRegister of Controlled Trials; WHO International Clinical Trials Registry Platform; conference proceedings; and reference lists to March 2014. Selection criteria Randomized and quasi-randomized controlled clinical trials in adults and children with cholera that compared: 1) any antimicrobial treatment with placebo or no treatment; 2) different antimicrobials head-to-head; or 3) different dosing schedules or different durations of treatment with the same antimicrobial. Data collection and analysis Two reviewers independently applied inclusion and exclusion criteria, and extracted data from included trials. Diarrhoea duration and stool volume were defined as primary outcomes. We calculated mean difference (MD) or ratio of means (ROM) for continuous outcomes, with 95% confidence intervals (CI), and pooled data using a random-effects meta-analysis. The quality of evidence was assessed using the GRADE approach. Main results Thirty-nine trials were included in this review with 4623 participants. Antimicrobials versus placebo or no treatment Overall, antimicrobial therapy shortened the mean duration of diarrhoea by about a day and a half compared to placebo or no treatment (MD -36.77 hours, 95% CI -43

  8. High case fatality cholera outbreak in Western Kenya, August 2010 ...

    African Journals Online (AJOL)

    Introduction: Cholera is a disease caused by the bacterium Vibrio cholera and has been an important public health problem since its first pandemic in 1817. Kenya has had numerous outbreaks of cholera ever since it was first detected there during 1971. In August 2010 an outbreak of cholera occurred in Kuria West District ...

  9. Intestinal Colonization Dynamics of Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    Salvador Almagro-Moreno

    2015-05-01

    Full Text Available To cause the diarrheal disease cholera, Vibrio cholerae must effectively colonize the small intestine. In order to do so, the bacterium needs to successfully travel through the stomach and withstand the presence of agents such as bile and antimicrobial peptides in the intestinal lumen and mucus. The bacterial cells penetrate the viscous mucus layer covering the epithelium and attach and proliferate on its surface. In this review, we discuss recent developments and known aspects of the early stages of V. cholerae intestinal colonization and highlight areas that remain to be fully understood. We propose mechanisms and postulate a model that covers some of the steps that are required in order for the bacterium to efficiently colonize the human host. A deeper understanding of the colonization dynamics of V. cholerae and other intestinal pathogens will provide us with a variety of novel targets and strategies to avoid the diseases caused by these organisms.

  10. Endoplasmic Reticulum-Targeted Subunit Toxins Provide a New Approach to Rescue Misfolded Mutant Proteins and Revert Cell Models of Genetic Diseases.

    Science.gov (United States)

    Adnan, Humaira; Zhang, Zhenbo; Park, Hyun-Joo; Tailor, Chetankumar; Che, Clare; Kamani, Mustafa; Spitalny, George; Binnington, Beth; Lingwood, Clifford

    2016-01-01

    Many germ line diseases stem from a relatively minor disturbance in mutant protein endoplasmic reticulum (ER) 3D assembly. Chaperones are recruited which, on failure to correct folding, sort the mutant for retrotranslocation and cytosolic proteasomal degradation (ER-associated degradation-ERAD), to initiate/exacerbate deficiency-disease symptoms. Several bacterial (and plant) subunit toxins, retrograde transport to the ER after initial cell surface receptor binding/internalization. The A subunit has evolved to mimic a misfolded protein and hijack the ERAD membrane translocon (dislocon), to effect cytosolic access and cytopathology. We show such toxins compete for ERAD to rescue endogenous misfolded proteins. Cholera toxin or verotoxin (Shiga toxin) containing genetically inactivated (± an N-terminal polyleucine tail) A subunit can, within 2-4 hrs, temporarily increase F508delCFTR protein, the major cystic fibrosis (CF) mutant (5-10x), F508delCFTR Golgi maturation (glucocerobrosidase (GCC) in N370SGCC Gaucher Disease fibroblasts (3x), another ERAD-exacerbated misfiling disease. We identify a new, potentially benign approach to the treatment of certain genetic protein misfolding diseases.

  11. Comparison of Escherichia coli Isolates from humans, food, and farm and companion animals for presence of Shiga toxin-producing E. coli virulence markers.

    Science.gov (United States)

    Murinda, Shelton E; Nguyen, Lien T; Landers, Tippi L; Draughon, F Ann; Mathew, Alan G; Hogan, Joseph S; Smith, K Larry; Hancock, Dale D; Oliver, Stephen P

    2004-01-01

    The objective of this study was to characterize Escherichia coli isolates from dairy cows/feedlots, calves, mastitis, pigs, dogs, parrot, iguana, human disease, and food products for prevalence of Shiga toxin-producing E. coli (STEC) virulence markers. The rationale of the study was that, isolates of the same serotypes that were obtained from different sources and possessed the same marker profiles, could be cross-species transmissible. Multiplex polymerase chain reaction (PCR) was used to detect presence of genes encoding Shiga toxin 1 and 2 (stx1 and stx2), H7 flagella (flicC), enterohemolysin (hly) and intimin (eaeA) in E. coli isolates (n = 400). Shiga toxin-producing isolates were tested for production of Shiga toxins (Stx1 and Stx2 and enterohemolysin. Of the E. coli O157:H7/H- strains, 150 of 164 (mostly human, cattle, and food) isolates were stx+. Sixty-five percent of O157 STEC produced both Stx1 and Stx2; 32% and 0.7% produced Stx2 or Stx1, respectively. Ninety-eight percent of O157 STEC had sequences for genes encoding intimin and enterohemolysin. Five of 20 E. coli O111, 4 of 14 O128 and 4 of 10 O26 were stx+ . Five of 6 stx+ O26 and O111 produced Stx1, however, stx+ O128 were Stx-negative. Acid resistance (93.3%) and tellurite resistance (87.3%) were common attributes of O157 STEC, whereas, non-O157 stx+ strains exhibited 38.5% and 30.8% of the respective resistances. stx-positive isolates were mostly associated with humans and cattle, whereas, all isolates from mastitis (n = 105), and pigs, dogs, parrot and iguanas (n = 48) were stx-negative. Multiplex PCR was an effective tool for characterizing STEC pathogenic profiles and distinguished STEC O157:H7 from other STEC. Isolates from cattle and human disease shared similar toxigenic profiles, whereas isolates from other disease sources had few characteristics in common with the former isolates. These data suggest interspecies transmissibility of certain serotypes, in particular, STEC O157:H7, between

  12. The Apoptotic Effects of the P300 Activator on Breast Cancer and Lung Fibroblast Cell Lines

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    Mohammad Reza Salahshoor

    2013-10-01

    Full Text Available Background: P300 is an enzyme that acetylates histones during stress. It alsoacetylates several non-histone proteins, including P53 which is the most important tumorsuppressor gene. P53 plays an important role in the apoptosis of tumor cells. Hereby,this study describes the potency of cholera toxin B subunit as a P300 activator to induceapoptosis in a breast cancer cell line (MCF-7 and a lung fibroblast cell line (MRC-5as a non-tumorigenic control sample. Methods: MCF-7 and MRC-5 were cultured in RPMI-1640 and treated with orwithout cholera toxin B subunit at the concentration of 85.43 μmol/L, based on the half-maximal inhibitory concentration index at different times (24, 48 and 72 h. Thepercentage of apoptotic cells was measured by flow cytometry. Real-time quantitativeRT-PCR was performed to estimate the mRNA expression of P300 in MCF-7 and MRC-5 with cholera toxin B subunit at different times. We used the ELISA and Bradford proteintechniques to detect levels of total and acetylated P53 protein generated in MCF-7 andMRC-5. Results: Our findings indicated that the cholera toxin B subunit effectively andsignificantly induced more apoptosis in MCF-7 compared to MRC-5. We showed thatexpression of P300 up-regulated by increasing the time of the cholera toxin B subunittreatment in MCF-7 but not in MRC-5. In addition, the acetylated and total P53protein levels increased more in MCF-7 cells than in MRC-5 cells.Conclusion: Cholera toxin B subunit induced significant cell death in MCF-7, butit could be well tolerated in MRC-5. Therefore, cholera toxin B subunit can besuggested as an anti-cancer agent.

  13. Cholera Vaccination Campaign Contributes to Improved Knowledge Regarding Cholera and Improved Practice Relevant to Waterborne Disease in Rural Haiti

    Science.gov (United States)

    Aibana, Omowunmi; Franke, Molly; Teng, Jessica; Hilaire, Johanne; Raymond, Max; Ivers, Louise C.

    2013-01-01

    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. PMID:24278498

  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 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.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.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. Local population and regional environmental drivers of cholera in Bangladesh

    Directory of Open Access Journals (Sweden)

    Escamilla Veronica

    2010-01-01

    Full Text Available Abstract Background Regional environmental factors have been shown to be related to cholera. Previous work in Bangladesh found that temporal patterns of cholera are positively related to satellite-derived environmental variables including ocean chlorophyll concentration (OCC. Methods This paper investigates whether local socio-economic status (SES modifies the effect of regional environmental forces. The study area is Matlab, Bangladesh, an area of approximately 200,000 people with an active health and demographic surveillance system. Study data include (1 spatially-referenced demographic and socio-economic characteristics of the population; (2 satellite-derived variables for sea surface temperature (SST, sea surface height (SSH, and OCC; and (3 laboratory confirmed cholera case data for the entire population. Relationships between cholera, the environmental variables, and SES are measured using generalized estimating equations with a logit link function. Additionally two separate seasonal models are built because there are two annual cholera epidemics, one pre-monsoon, and one post-monsoon. Results SES has a significant impact on cholera occurrence: the higher the SES score, the lower the occurrence of cholera. There is a significant negative association between cholera incidence and SSH during the pre-monsoon period but not for the post-monsoon period. OCC is positively associated with cholera during the pre-monsoon period but not for the post-monsoon period. SST is not related to cholera incidence. Conclusions Overall, it appears cholera is influenced by regional environmental variables during the pre-monsoon period and by local-level variables (e.g., water and sanitation during the post-monsoon period. In both pre- and post-monsoon seasons, SES significantly influences these patterns, likely because it is a proxy for poor water quality and sanitation in poorer households.

  16. A novel kit for rapid detection of Vibrio cholerae O1.

    Science.gov (United States)

    Hasan, J A; Huq, A; Tamplin, M L; Siebeling, R J; Colwell, R R

    1994-01-01

    We report on the development and testing of a novel, rapid, colorimetric immunodiagnostic kit, Cholera SMART, for direct detection of the presence of Vibrio cholerae O1 in clinical specimens. Unlike conventional culture methods requiring several days to complete, the Cholera SMART kit can be used directly in the field by untrained or minimally skilled personnel to detect V. cholerae O1 in less than 15 min, without cumbersome laboratory equipment. A total of 120 clinical and environmental bacterial strains, including both O1 and non-O1 serotypes of V. cholerae isolated from samples collected from a variety of geographical regions, were tested, and positive reactions were observed only with V. cholerae O1. Also, results of a field trial in Bangladesh, employing Cholera SMART, showed 100% specificity and 96% sensitivity compared with conventional culture methods. Another field trial, in Mexico, showed that Cholera SMART was 100% in agreement with a recently described coagglutination test when 108 stool specimens were tested.

  17. Adapting to the global shortage of cholera vaccines: targeted single dose cholera vaccine in response to an outbreak in South Sudan.

    Science.gov (United States)

    Parker, Lucy A; Rumunu, John; Jamet, Christine; Kenyi, Yona; Lino, Richard Laku; Wamala, Joseph F; Mpairwe, Allan M; Ciglenecki, Iza; Luquero, Francisco J; Azman, Andrew S; Cabrol, Jean-Clement

    2017-04-01

    Shortages of vaccines for epidemic diseases, such as cholera, meningitis, and yellow fever, have become common over the past decade, hampering efforts to control outbreaks through mass reactive vaccination campaigns. Additionally, various epidemiological, political, and logistical challenges, which are poorly documented in the literature, often lead to delays in reactive campaigns, ultimately reducing the effect of vaccination. In June 2015, a cholera outbreak occurred in Juba, South Sudan, and because of the global shortage of oral cholera vaccine, authorities were unable to secure sufficient doses to vaccinate the entire at-risk population-approximately 1 million people. In this Personal View, we document the first public health use of a reduced, single-dose regimen of oral cholera vaccine, and show the details of the decision-making process and timeline. We also make recommendations to help improve reactive vaccination campaigns against cholera, and discuss the importance of new and flexible context-specific dose regimens and vaccination strategies. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Use of oral cholera vaccine as a vaccine probe to define the geographical dimensions of person-to-person transmission of cholera.

    Science.gov (United States)

    Ali, Mohammad; Kim, Deok Ryun; Kanungo, Suman; Sur, Dipika; Manna, Byomkesh; Digilio, Laura; Dutta, Shanta; Marks, Florian; Bhattacharya, Sujit K; Clemens, John

    2018-01-01

    Cholera is known to be transmitted from person to person, and inactivated oral cholera vaccines (OCVs) have been shown to confer herd protection via interruption of this transmission. However, the geographic dimensions of chains of person-to-person transmission of cholera are uncertain. The ability of OCVs to confer herd protection was used to define these dimensions in two cholera-endemic settings, one in rural Bangladesh and the other in urban India. Two large randomized, placebo-controlled trials of inactivated OCVs, one in rural Matlab, Bangladesh and the other in urban Kolkata, India, were reanalyzed. Vaccine herd protection was evaluated by relating the risk of cholera in placebo recipients to vaccine coverage of surrounding residents residing within concentric rings. In Matlab, concentric rings in 100-m increments up to 700m were evaluated; in Kolkata, 50-m increments up to 350m were evaluated. One hundred and eight cholera cases among 24667 placebo recipients were detected during 1year of post-vaccination follow-up at Matlab; 128 cholera cases among 34968 placebo recipients were detected during 3 years of follow-up in Kolkata. Consistent inverse relationships were observed between vaccine coverage of the ring and the risk of cholera in the central placebo recipient for rings with radii up to 500m in Matlab and up to 150m in Kolkata. These results suggest that the dimensions of chains of person-to-person transmission in endemic settings can be quite large and may differ substantially from setting to setting. Using OCVs as 'probes' to define these dimensions can inform geographical targeting strategies for the deployment of these vaccines in endemic settings. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

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

    NARCIS (Netherlands)

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

    2012-01-01

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

  20. Cholera in travelers: shifting tides in epidemiology, management, and prevention.

    Science.gov (United States)

    Fillion, Katie; Mileno, Maria D

    2015-01-01

    The distribution of cholera's devastating effects has changed. While cholera is endemic in 50 countries mostly in Asia and Africa, more than half of the cases reported in 2012 were in the Western Hemisphere, predominantly Haiti. Since the current epidemic began in Haiti in 2010, there has been spread to the Dominican Republic, Cuba, and most recently Mexico. Several recent case reports document individuals returning home from affected areas with diarrhea from cholera, in some cases severe. Hopeful news reported the containment of an outbreak through the use of a Vibrio cholera vaccine. There are safe and effective oral cholera vaccines available and recommended in outbreaks and endemic areas, although they are not currently available in the USA or to travelers. This review aims to discuss the latest data to aid our current recommendations for the prevention of cholera in travelers beyond standard personal and food hygiene precautions for the prevention of travelers' diarrhea and to offer insights on the most current data available about cholera vaccine progress and potential use.

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

    Directory of Open Access Journals (Sweden)

    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.

  2. Cholera Epidemic - Lusaka, Zambia, October 2017-May 2018.

    Science.gov (United States)

    Sinyange, Nyambe; Brunkard, Joan M; Kapata, Nathan; Mazaba, Mazyanga Lucy; Musonda, Kunda G; Hamoonga, Raymond; Kapina, Muzala; Kapaya, Fred; Mutale, Lwito; Kateule, Ernest; Nanzaluka, Francis; Zulu, James; Musyani, Chileshe Lukwesa; Winstead, Alison V; Davis, William W; N'cho, Hammad S; Mulambya, Nelia L; Sakubita, Patrick; Chewe, Orbie; Nyimbili, Sulani; Onwuekwe, Ezinne V C; Adrien, Nedghie; Blackstock, Anna J; Brown, Travis W; Derado, Gordana; Garrett, Nancy; Kim, Sunkyung; Hubbard, Sydney; Kahler, Amy M; Malambo, Warren; Mintz, Eric; Murphy, Jennifer; Narra, Rupa; Rao, Gouthami G; Riggs, Margaret A; Weber, Nicole; Yard, Ellen; Zyambo, Khozya D; Bakyaita, Nathan; Monze, Namani; Malama, Kennedy; Mulwanda, Jabbin; Mukonka, Victor M

    2018-05-18

    On October 6, 2017, an outbreak of cholera was declared in Zambia after laboratory confirmation of Vibrio cholerae O1, biotype El Tor, serotype Ogawa, from stool specimens from two patients with acute watery diarrhea. The two patients had gone to a clinic in Lusaka, the capital city, on October 4. Cholera cases increased rapidly, from several hundred cases in early December 2017 to approximately 2,000 by early January 2018 (Figure). In collaboration with partners, the Zambia Ministry of Health (MoH) launched a multifaceted public health response that included increased chlorination of the Lusaka municipal water supply, provision of emergency water supplies, water quality monitoring and testing, enhanced surveillance, epidemiologic investigations, a cholera vaccination campaign, aggressive case management and health care worker training, and laboratory testing of clinical samples. In late December 2017, a number of water-related preventive actions were initiated, including increasing chlorine levels throughout the city's water distribution system and placing emergency tanks of chlorinated water in the most affected neighborhoods; cholera cases declined sharply in January 2018. During January 10-February 14, 2018, approximately 2 million doses of oral cholera vaccine were administered to Lusaka residents aged ≥1 year. However, in mid-March, heavy flooding and widespread water shortages occurred, leading to a resurgence of cholera. As of May 12, 2018, the outbreak had affected seven of the 10 provinces in Zambia, with 5,905 suspected cases and a case fatality rate (CFR) of 1.9%. Among the suspected cases, 5,414 (91.7%), including 98 deaths (CFR = 1.8%), occurred in Lusaka residents.

  3. Efficiency of hospital cholera treatment in Ecuador

    Directory of Open Access Journals (Sweden)

    Creamer Germán

    1999-01-01

    Full Text Available This study analyzed the efficiency of cholera treatment in three hospitals representative of the Ecuadorian public health system in order to provide hospital directors and administrators and health service policy-makers with information to plan responses to future epidemics and to reduce the costs of cholera treatment in general. For the study, total and excess cholera treatment costs were calculated using hospital files and statistics and an in-hospital surveillance system of the cholera cases. The type and quantity of each input used for each treatment were analyzed, as well as the number of days hospitalized, according to the severity of the illness. With this process, excess costs were determined in relation to a "treatment norm" that would have been appropriate for each patient. The researchers found that 45% of the cholera treatment costs were excessive. The most important contributor was excess recurrent costs (90%, including extended hospital stays, disproportionate use of intravenous rehydration solutions, and unnecessary laboratory tests. Excess capital costs, from land, buildings, and hospital equipment, represented 10% of the total excess treatment costs. No significant relationship was found between treatment costs and the severity of the illness, nor between costs and a patient's age. A patient's sex appeared to be an important variable, with the cost of treating women being notably higher than for men. An inverse relationship was found between treatment costs and the complexity of the hospital. The researchers concluded there was an inefficient use of resources in the treatment of cholera in the three hospitals where the research was performed.

  4. Requirement for Vibrio cholerae integration host factor in conjugative DNA transfer.

    Science.gov (United States)

    McLeod, Sarah M; Burrus, Vincent; Waldor, Matthew K

    2006-08-01

    The requirement for host factors in the transmission of integrative and conjugative elements (ICEs) has not been extensively explored. Here we tested whether integration host factor (IHF) or Fis, two host-encoded nucleoid proteins, are required for transfer of SXT, a Vibrio cholerae-derived ICE that can be transmitted to many gram-negative species. Fis did not influence the transfer of SXT to or from V. cholerae. In contrast, IHF proved to be required for V. cholerae to act as an SXT donor. In the absence of IHF, V. cholerae displayed a modest defect for serving as an SXT recipient. Surprisingly, SXT integration into or excision from the V. cholerae chromosome, which requires an SXT-encoded integrase related to lambda integrase, did not require IHF. Therefore, the defect in SXT transmission in the V. cholerae IHF mutant is probably not related to IHF's ability to promote DNA recombination. The V. cholerae IHF mutant was also highly impaired as a donor of RP4, a broad-host-range conjugative plasmid. Thus, the V. cholerae IHF mutant appears to have a general defect in conjugation. Escherichia coli IHF mutants were not impaired as donors or recipients of SXT or RP4, indicating that IHF is a V. cholerae-specific conjugation factor.

  5. Hydroclimatic Extremes and Cholera Dynamics in the 21st Century

    Science.gov (United States)

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

    2012-12-01

    Cholera, an acute water-borne diarrheal illness, has reemerged as a significant health threat across much of the developing world. Despite major advances in the ecological and the microbiological understanding of the causative agent, V. cholerae, the role of the underlying climatic and environmental processes in propagating transmission is not adequately understood. Recent findings suggest a more prominent role of hydroclimatic extremes - droughts and floods - on the unique dual cholera peaks in the Bengal Delta region of South Asia, the native homeland of cholera. Increasing water scarcity and abundance, and coastal sea-level rise, influenced by changing climate patterns and large-scale climatic phenomena, is likely to adversely impact cholera transmission in South Asia. We focus on understanding how associated changes in macro-scale conditions in this region will impact micro-scale processes related to cholera in coming decades. We use the PRECIS Regional Climate Model over the Ganges-Brahmaputra-Meghna (GBM) basin region to simulate detailed high resolution projections of climate patterns for the 21st century. Precipitation outputs are analyzed for the 1980-2040 period to identify the trends and changes in hydroclimatic extremes and potential impacts on cholera dynamics over the next three decades (2010-2040), in relation to the cholera surveillance operations over the past three decades (1980-2010). We find that an increased number of extreme precipitation events with prolonged dry periods in the Ganges basin region will likely adversely affect dry season cholera outbreaks. Increased monsoon precipitation volumes in the Brahmaputra basin catchments are likely to cause record floods and subsequently trigger large epidemics in downstream areas. Our results provide new insight by identifying the changes in the two distinctly different, pre and post monsoon, cholera transmission mechanisms related to large-scale climatic controls that prevail in the region. A

  6. [Antibiotic resistance pattern of 24, 526 strains of Vibrio cholerae O1 isolated in Mexico from 1991 to 1993].

    Science.gov (United States)

    Giono-Cerezo, S; Zárate, A; Gutiérrez, L; Valdespino, J L

    1994-01-01

    Profile of antimicrobial resistance by Kirby-Bauer method was performed on 24526 Vibrio cholerae O1 strains isolated in México (1991-1993) from fecal swabs in cholera cases and from asymptomatic carriers. Minimal inhibitory concentration (MIC) tests for tetracycline (Te) and doxycycline (D) were done on selected strains. Single antibiotic discs were used at concentrations of: Te, 30 micrograms; D, 30 micrograms; erythromycin (E), 15 micrograms; chloramphenicol (CM), 30 micrograms; ampicillin (AM), 10 micrograms; trimethoprim-sulfamethoxazole (SXT) 1.25 micrograms/23.75 micrograms. Strains whose halos were of a smaller diameter than the intermediate value were considered resistant. It is important to maintain surveillance on antimicrobial susceptibility as epidemiological marker on geographical selected areas in order to detect changes of resistant patterns.

  7. Estimating effects of improved drinking water and sanitation on cholera.

    Science.gov (United States)

    Leidner, Andrew J; Adusumilli, Naveen C

    2013-12-01

    Demand for adequate provision of drinking-water and sanitation facilities to promote public health and economic growth is increasing in the rapidly urbanizing countries of the developing world. With a panel of data on Asia and Africa from 1990 to 2008, associations are estimated between the occurrence of cholera outbreaks, the case rates in given outbreaks, the mortality rates associated with cholera and two disease control mechanisms, drinking-water and sanitation services. A statistically significant and negative effect is found between drinking-water services and both cholera case rates as well as cholera-related mortality rates. A relatively weak statistical relationship is found between the occurrence of cholera outbreaks and sanitation services.

  8. Rare codons effect on expression of recombinant gene cassette in Escherichia coli BL21(DE3

    Directory of Open Access Journals (Sweden)

    Aghil Esmaeili-Bandboni

    2017-11-01

    Full Text Available Objective: To demonstrate the sensitivity of expression of fusion genes to existence of a large number of rare codons in recombinant gene sequenced. Methods: Primers for amplification of cholera toxin B, Shiga toxin B and gfp genes were designed by Primer3 software and synthesized. All of these 3 genes were cloned. Then the genes were fused together by restriction sites and enzymatic method. Two linkers were used as a flexible bridge in connection of these genes. Results: Cloning and fusion of cholera toxin B, Shiga toxin B and gfp genes were done correctly. After that, expression of the recombinant gene construction was surveyed. Conclusions: According to what was seen, because of the accumulation of 12 rare codons of Shiga toxin B and 19 rare codons of cholera toxin B in this gene cassette, the expression of the recombinant gene cassette, in Escherichia coli BL21, failed.

  9. Influence of human behavior on cholera dynamics.

    Science.gov (United States)

    Wang, Xueying; Gao, Daozhou; Wang, Jin

    2015-09-01

    This paper is devoted to studying the impact of human behavior on cholera infection. We start with a cholera ordinary differential equation (ODE) model that incorporates human behavior via modeling disease prevalence dependent contact rates for direct and indirect transmissions and infectious host shedding. Local and global dynamics of the model are analyzed with respect to the basic reproduction number. We then extend the ODE model to a reaction-convection-diffusion partial differential equation (PDE) model that accounts for the movement of both human hosts and bacteria. Particularly, we investigate the cholera spreading speed by analyzing the traveling wave solutions of the PDE model, and disease threshold dynamics by numerically evaluating the basic reproduction number of the PDE model. Our results show that human behavior can reduce (a) the endemic and epidemic levels, (b) cholera spreading speeds and (c) the risk of infection (characterized by the basic reproduction number). Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Role of phytoplankton in maintaining endemicity and seasonality of cholera in Bangladesh.

    Science.gov (United States)

    Islam, M Sirajul; Islam, M Shafiqul; Mahmud, Zahid H; Cairncross, Sandy; Clemens, John D; Collins, Andrew E

    2015-09-01

    In Bangladesh, cholera is endemic and maintains a regular seasonal pattern. The role of phytoplankton in maintaining endemicity and seasonality of cholera was monitored in Matlab, Bangladesh. Phytoplankton and water samples were collected from two ponds bi-weekly for 1 year. The association of Vibrio cholerae O1 with phytoplankton was studied by culture and direct fluorescent antibody techniques. The bio-physicochemical parameters of water were measured and data for cases of cholera were collected from the records of Matlab hospital. The correlation of cholera cases with levels of phytoplankton, V. cholerae and bio-physicochemical parameters of water was carried out using Pearson's correlation coefficients. V. cholerae O1 survived for 48 days in association with Anabaena variabilis in a culturable state, but survived for a year in a viable but non-culturable (VBNC) state. V. cholerae survived for 12 and 32 days in a culturable state in control water (without algae) and water with algae, respectively. There was a significant correlation between changing levels of cholera cases in the community and the blue green algae and total phytoplankton in the aquatic environment. A significant correlation was also found between the cholera cases and chlorophyll-a and VBNC V. cholerae O1 in the aquatic environment. This study demonstrated the role of phytoplankton in maintaining endemicity and seasonality of cholera in Bangladesh. © The Author 2015. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Cholera dynamics with Bacteriophage infection: A mathematical study

    International Nuclear Information System (INIS)

    Misra, A.K.; Gupta, Alok; Venturino, Ezio

    2016-01-01

    Highlights: • A mathematical model for the biological control of cholera has been proposed. • The feasibility and stability of all the equilibria have been investigated. • The ODE model is found to exhibit Hopf-bifurcation. • Conditions of global asymptotic stability have been obtained. • The impact of important parameters on cholera spread has been shown. - Abstract: Mathematical modeling of waterborne diseases, such as cholera, including a biological control using Bacteriophage viruses in the aquatic reservoirs is of great relevance in epidemiology. In this paper, our aim is twofold: at first, to understand the cholera dynamics in the region around a water body; secondly, to understand how the spread of Bacteriophage infection in the cholera bacterium V. cholerae controls the disease in the human population. For this purpose, we modify the model proposed by Codeço, for the spread of cholera infection in human population and the one proposed by Beretta and Kuang, for the spread of Bacteriophage infection in the bacteria population [1, 2]. We first discuss the feasibility and local asymptotic stability of all the possible equilibria of the proposed model. Further, in the numerical investigation, we have found that the parameter ϕ, called the phage adsorption rate, plays an important role. There is a critical value, ϕ c , at which the model possess Hopf-bifurcation. For lower values than ϕ c , the equilibrium E * is unstable and periodic solutions are observed, while above ϕ c , the equilibrium E * is locally asymptotically stable, and further shown to be also globally asymptotically stable. We investigate the effect of the various parameters on the dynamics of the infected humans by means of numerical simulations.

  12. The Orphans of Cholera Morbus in Yucatan, 1833

    OpenAIRE

    Elsa Malvido; Paola Peniche Moreno

    2013-01-01

    This essay discusses the phenomenon of orphanhood which affected a large number of children after the cholera epidemic that struck Yucatan in July 1833. Moreover, it inquires into the fate of children whose parents died of cholera, the role played by kinship networks to provide them with shelter, and the influence of the Church and the State on the situation. Based on first hand sources, the author suggests that the orphanhood produced by cholera served as a pretext for economically and socia...

  13. Zonula occludens toxin structure-function analysis. Identification of the fragment biologically active on tight junctions and of the zonulin receptor binding domain.

    Science.gov (United States)

    Di Pierro, M; Lu, R; Uzzau, S; Wang, W; Margaretten, K; Pazzani, C; Maimone, F; Fasano, A

    2001-06-01

    Zonula occludens toxin (Zot) is an enterotoxin elaborated by Vibrio cholerae that increases intestinal permeability by interacting with a mammalian cell receptor with subsequent activation of intracellular signaling leading to the disassembly of the intercellular tight junctions. Zot localizes in the bacterial outer membrane of V. cholerae with subsequent cleavage and secretion of a carboxyl-terminal fragment in the host intestinal milieu. To identify the Zot domain(s) directly involved in the protein permeating effect, several zot gene deletion mutants were constructed and tested for their biological activity in the Ussing chamber assay and their ability to bind to the target receptor on intestinal epithelial cell cultures. The Zot biologically active domain was localized toward the carboxyl terminus of the protein and coincided with the predicted cleavage product generated by V. cholerae. This domain shared a putative receptor-binding motif with zonulin, the Zot mammalian analogue involved in tight junction modulation. Amino acid comparison between the Zot active fragment and zonulin, combined with site-directed mutagenesis experiments, confirmed the presence of an octapeptide receptor-binding domain toward the amino terminus of the processed Zot.

  14. Twee Nederlandse reizigers uit Thailand met cholera

    NARCIS (Netherlands)

    Smit, A. A.; Kuijper, E. J.; Schultz, M. J.; Wieling, W.; Speelman, P.

    1994-01-01

    Cholera is a disease rarely imported in the Netherlands. Recently a 34-year-old woman who had returned from a trip through Thailand was admitted to our hospital with complaints of vomiting, watery stools and moderate dehydration. Vibrio cholerae OI serotype Ogawa biotype El Tor was isolated from the

  15. Epidemic cholera in rural El Salvador: risk factors in a region covered by a cholera prevention campaign.

    Science.gov (United States)

    Quick, R E; Thompson, B L; Zuniga, A; Dominguez, G; De Brizuela, E L; De Palma, O; Almeida, S; Valencia, A; Ries, A A; Bean, N H

    1995-04-01

    In response to the Latin American cholera epidemic, El Salvador began a prevention programme in April 1991. The first case was confirmed in August, and 700 cases were reported within 3 months. A matched case-control study was conducted in rural La Libertad Department in November 1991. Illness was associated with eating cold cooked or raw seafood (odds ratio [OR] = 7.0; 95% confidence limits [CL] = 1.4, 35.0) and with drinking water outside the home (OR = 8.8; 95% CL = 1.7, 44.6). Assertion of knowledge about how to prevent cholera (OR = 0.2; 95% CL = 0.1, 0.8) and eating rice (OR = 0.2; 95% CL = 0.1, 0.8) were protective. More controls than patients regularly used soap (OR = 0.3; 95% CL = 0.1, 1.0). This study demonstrated three important points for cholera prevention: (1) seafood should be eaten cooked and hot; (2) populations at risk should be taught to treat household drinking water and to avoid drinking water outside the home unless it is known to be treated; and (3) education about hygiene can be an important tool in preventing cholera.

  16. Vibrio cholerae Classical Biotype Strains Reveal Distinct Signatures in Mexico

    OpenAIRE

    Alam, Munirul; Islam, M. Tarequl; Rashed, Shah Manzur; Johura, Fatema-tuz; Bhuiyan, Nurul A.; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Watanabe, Haruo; Hasan, Nur-A; Colwell, Rita R.; Cravioto, Alejandro

    2012-01-01

    Vibrio cholerae O1 classical (CL) biotype caused the fifth and sixth pandemics, and probably the earlier cholera pandemics, before the El Tor (ET) biotype initiated the seventh pandemic in Asia in the 1970s by completely displacing the CL biotype. Although the CL biotype was thought to be extinct in Asia and although it had never been reported from Latin America, V. cholerae CL and ET biotypes, including a hybrid ET, were found associated with areas of cholera endemicity in Mexico between 199...

  17. 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.

  18. What is Cholera?

    Indian Academy of Sciences (India)

    Cholera is a rapidly dehydrating watery diarrheal disease that can lead to death in less than 24 hours if untreated, making it, according to WHO, “one of the most rapidly fatal infectious illnesses known” ...

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

    Directory of Open Access Journals (Sweden)

    Daniel Rios Garza

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

  20. Non-01 Vibrio cholerae infections in Cancun, Mexico.

    Science.gov (United States)

    Finch, M J; Valdespino, J L; Wells, J G; Perez-Perez, G; Arjona, F; Sepulveda, A; Bessudo, D; Blake, P A

    1987-03-01

    To determine the role of Vibrio cholerae as a cause of diarrheal illness in Cancun, Mexico, an investigation was conducted in July and August 1983. Although toxigenic V. cholerae 01 were not found, non-01 V. cholerae were isolated from 22 (16%) of 134 stools from persons with diarrheal illness and none of 22 stools from well persons; 58 (92%) of 63 sewage samples; 12 (86%) of 14 untreated well water samples; a home storage tank for treated water; and 5 (21%) of 24 samples of raw seafood. None of the V. cholerae isolates from patients were toxigenic. The illness occurred mainly in small children, and were characterized principally by diarrhea and abdominal pain. No patient was seriously ill, and all recovered without sequelae. Seven different serotypes of non-01 V. cholerae were isolated from the stool specimens, and Smith serotype 12 accounted for 10 (46%) of the 22 isolates. A matched-pair case-control study found that cases were more likely than controls to have eaten home prepared gelatin (P = 0.03, OR = 5/0) and seafood (P = 0.06, OR = 4/0).

  1. Seasonality of cholera from 1974 to 2005: a review of global patterns

    Directory of Open Access Journals (Sweden)

    Feldacker Caryl

    2008-06-01

    Full Text Available Abstract Background The seasonality of cholera is described in various study areas throughout the world. However, no study examines how temporal cycles of the disease vary around the world or reviews its hypothesized causes. This paper reviews the literature on the seasonality of cholera and describes its temporal cycles by compiling and analyzing 32 years of global cholera data. This paper also provides a detailed literature review on regional patterns and environmental and climatic drivers of cholera patterns. Data, Methods, and Results Cholera data are compiled from 1974 to 2005 from the World Health Organization Weekly Epidemiological Reports, a database that includes all reported cholera cases in 140 countries. The data are analyzed to measure whether season, latitude, and their interaction are significantly associated with the country-level number of outbreaks in each of the 12 preceding months using separate negative binomial regression models for northern, southern, and combined hemispheres. Likelihood ratios tests are used to determine the model of best fit. The results suggest that cholera outbreaks demonstrate seasonal patterns in higher absolute latitudes, but closer to the equator, cholera outbreaks do not follow a clear seasonal pattern. Conclusion The findings suggest that environmental and climatic factors partially control the temporal variability of cholera. These results also indirectly contribute to the growing debate about the effects of climate change and global warming. As climate change threatens to increase global temperature, resulting rises in sea levels and temperatures may influence the temporal fluctuations of cholera, potentially increasing the frequency and duration of cholera outbreaks.

  2. A simple and convenient microtiter plate assay for the detection of bactericidal antibodies to Vibrio cholerae O1 and Vibrio cholerae O139.

    Science.gov (United States)

    Boutonnier, Alain; Dassy, Bruno; Duménil, Rémy; Guénolé, Alain; Ratsitorahina, Maherisoa; Migliani, René; Fournier, Jean-Michel

    2003-12-01

    It is believed that the correlate of protection for cholera can be determined by the serum vibriocidal assay. The currently available vibriocidal assays, based on the conventional agar plating technique, are labor intensive. We developed a simple and convenient microtiter plate assay for the detection of vibriocidal antibodies that is equally as efficient for Vibrio cholerae O1 and for V. cholerae O139. The addition of succinate and neotetrazolium made it possible to measure the growth of surviving bacterial target cells by monitoring a color change. We evaluated assay parameters (target strains, growth of target cells, complement source and concentration) that may affect the reproducibility of the method for V. cholerae O139. The results obtained with the microtiter plate assay were uniformly similar to those obtained with the conventional agar plating assay, when testing both the Inaba and Ogawa serotypes of V. cholerae O1. The microtiter plate assay was also convenient for measuring the activity of animal sera and mouse monoclonal antibodies.

  3. Salovum egg yolk containing antisecretory factor as an adjunct therapy in severe cholera in adult males: a pilot study.

    Science.gov (United States)

    Alam, Nur H; Ashraf, Hasan; Olesen, Maryam; Salam, Mohammed A; Gyr, Niklaus; Meier, Remy

    2011-08-01

    Cholera involves stimulation of intestinal secretory process in response to cholera toxin leading to profuse watery diarrhoea that might cause death due to dehydration unless timely rehydration therapy is initiated. Efforts to identify and test potential antisecretory agents are ongoing. Antisecretory factor (AF) is a naturally-occurring protein produced in the human secretory organs, including the intestine, with antisectory properties demonstrated in animal and human models of secretory diarrhoea. Salovum egg yolk powder contains antisecretory proteins in a much higher (500 times) concentration than that of normal hen eggs. This is achieved by feeding hens with specially-processed cereals, capable of inducing antisecretory proteins in the yolk. The aim of the study was to examine the effect of Salovum egg yolk powder containing AF in the treatment of adult cholera patients. In an open, randomized controlled trial (pilot study), 40 adult male patients with severe cholera were studied: 20 received standard treatment (oral rehydration solution, antibiotic, and usual hospital diet) plus Salovum egg yolk powder (study group) and 20 received standard treatment alone (control group). All the patients received tablet doxycycline (300 mg) once immediately after randomization. Written informed consent was obtained from each subject before enrollment. The main outcome measures were stool weight and duration of diarrhoea. The demographic and baseline clinical characteristics of the study patients were comparable between the groups. No significant differences were found in the mean stool weight, g/kg of body-weight during the first 24 hours [study vs control group, mean +/- standard deviation (SD), 218 +/- 119 vs 195 +/- 136], second 24 hours (mean +/- SD, 23 +/- 39 vs 22 +/- 34), and cumulative up to 72 hours (mean +/- SD, 245 +/- 152 vs 218 +/- 169). The duration (hours) of diarrhoea after admission in the hospital was also similar in both the groups (mean +/- SD, 33 +/- 14

  4. Health impairments arising from drinking water resources contaminated with Vibrio cholerae.

    Science.gov (United States)

    Ramamurthy, T; Chakraborty, S; Nair, G B; Bhattacharya, S K

    2000-01-01

    The endemic and seasonal nature of cholera depends upon the survival of toxigenic Vibrio cholerae in various niches of the aquatic environment. To understand the transmission and ecology of V. cholerae, it is necessary to know which component in the aquatic ecosystem can harbor it and thus contribute to the endemic presence. Toxigenic V. cholerae is now recognized as an autochthonous member of the microflora in many aquatic environments based on its protracted survival and proliferation without losing the virulence determinants. This article summarizes knowledge about the ecology, survival strategies and elimination techniques of V. cholerae from natural waters with special reference to drinking water.

  5. 9 CFR 309.5 - Swine; disposal because of hog cholera.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Swine; disposal because of hog cholera... INSPECTION AND CERTIFICATION ANTE-MORTEM INSPECTION § 309.5 Swine; disposal because of hog cholera. (a) All swine found by an inspector to be affected with hog cholera shall be identified as U.S. Condemned and...

  6. Breast milk reduces the risk of illness in children of mothers with cholera

    DEFF Research Database (Denmark)

    Qureshi, Katja; Mølbak, Kåre; Sandström, Anita

    2006-01-01

    BACKGROUND: A protective effect of breastfeeding against cholera has been demonstrated in areas endemic of cholera. To assess the protection offered by breast milk from mothers living in an area that had been free from cholera for 7 years, we investigated mothers with cholera and their children...... during an epidemic with Vibrio cholerae El Tor in the capital of Guinea-Bissau. METHODS: Eighty mothers with clinical cholera and their children were identified, and interviewed. Blood samples for vibriocidal and antitoxin antibodies were collected from mother-and-child pairs. Breast milk samples were...... collected from lactating mothers.Cholera was defined as acute watery diarrhea during the epidemic and a vibriocidal reciprocal titer of 20 or above. RESULTS: Three (7%) of 42 breastfed children had cholera as defined above compared with 9 (24%) of 38 nonbreastfed children (RR for breastfed children, 0...

  7. Recommendations of the Advisory Committee on Immunization Practices for Use of Cholera Vaccine.

    Science.gov (United States)

    Wong, Karen K; Burdette, Erin; Mahon, Barbara E; Mintz, Eric D; Ryan, Edward T; Reingold, Arthur L

    2017-05-12

    Cholera, caused by infection with toxigenic Vibrio cholerae bacteria of serogroup O1 (>99% of global cases) or O139, is characterized by watery diarrhea that can be severe and rapidly fatal without prompt rehydration. Cholera is endemic in approximately 60 countries and causes epidemics as well. Globally, cholera results in an estimated 2.9 million cases of disease and 95,000 deaths annually (1). Cholera is rare in the United States, and most U.S. cases occur among travelers to countries where cholera is endemic or epidemic. Forty-two U.S. cases were reported in 2011 after a cholera epidemic began in Haiti (2); however, <25 cases per year have been reported in the United States since 2012.

  8. Coliform and Vibrio cholerae Analysis of Drinking Water Collected from Cholera Outbreak Region of Bhaktapur Municipality

    Directory of Open Access Journals (Sweden)

    Upendra Thapa Shrestha

    2014-09-01

    Full Text Available Water borne infections in Nepal, especially in Kathmandu valley is one the major public health problems, causing thousands of deaths every year. Among three cities in the valley, the water borne infection including cholera is most predominant in Bhaktapur district. So the study was carried out to know the microbial drinking water quality in the city and to determine the prevalence of water borne infections in the specified region of the district in 2012. Altogether eighty (two samples from a single site at different interval-2/3 days water samples were collected from Bhaktapur Municipality, one of the most vulnerable regions for water borne diseases, following standard methods as described by APHA, 2010. All samples were transferred to Microbiology laboratory of Khwopa College, Dekocha, Bhaktapur and preceded immediately for Microbial analysis. The coliform density in the water samples were determined by Most Probable Number (MPN method followed by microscopy, colonial morphology and biochemical characterization. Subsequently, the presence of Vibrio cholerae, a causative agent of Cholera was analyzed in the same samples by enrichment in alkaline peptone water followed by culture on Thiosulphate citrate bile-salt sucrose (TCBS agar, a selective media for Vibrio spp. The biochemical tests were then performed to identify V. cholerae. Among eighty water samples, 87.5 percent water samples contained coliforms and half of which (45% contained feacal coliforms, Escherichia coli and remaining 12.5 percent water samples contained no coliforms. Vibrio cholerae were isolated from four water samples (5%. The drinking water quality in the region was found to be very poor. Therefore, the people in the region were suggested to treat the drinking water by using any of physical or chemical disinfection methods prior to drinking. DOI: http://dx.doi.org/10.3126/ije.v3i3.11073 International Journal of Environment Vol.3(3 2014: 139-145

  9. [A prognostic model of a cholera epidemic].

    Science.gov (United States)

    Boev, B V; Bondarenko, V M; Prokop'eva, N V; San Román, R T; Raygoza-Anaya, M; García de Alba, R

    1994-01-01

    A new model for the prognostication of cholera epidemic on the territory of a large city is proposed. This model reflects the characteristic feature of contacting infection by sensitive individuals due to the preservation of Vibrio cholerae in their water habitat. The mathematical model of the epidemic quantitatively reflects the processes of the spread of infection by kinetic equations describing the interaction of the streams of infected persons, the causative agents and susceptible persons. The functions and parameters of the model are linked with the distribution of individuals according to the duration of the incubation period and infectious process, as well as the period of asymptomatic carrier state. The computer realization of the model by means of IBM PC/AT made it possible to study the cholera epidemic which took place in Mexico in 1833. The verified model of the cholera epidemic was used for the prognostication of the possible spread of this infection in Guadalajara, taking into account changes in the epidemiological situation and the size of the population, as well as improvements in sanitary and hygienic conditions, in the city.

  10. El Niño, Rainfall, and the Shifting Geography of Cholera in Africa

    Science.gov (United States)

    Moore, S.; Azman, A. S.; Zaitchik, B. F.; McKay, H.; Lessler, J.

    2017-12-01

    The El Niño Southern Oscillation (ENSO) and other climate patterns can have profound impacts on the occurrence of infectious diseases. Because of the key role of water supplies in cholera transmission, a relationship between El Niño events and cholera incidence is highly plausible, and previous research has shown a link between El Niño patterns and cholera in Bangladesh. However, there is little systematic evidence for this link in Africa where many cholera cases and deaths are reported. To understand how ENSO affects the geographic distribution of cholera incidence in Africa, we used a hierarchical Bayesian approach to integrate over 17,000 annual observations of cholera incidence from 2000-2014 in over 3,000 unique locations of varying spatial extent, ranging from entire countries to neighborhoods. The resulting maps reflect modeled cholera incidence at a fine spatial resolution using reported counts of cholera cases, key explanatory variables, and a spatially-dependent covariance term. We then examined the potential mechanistic association between ENSO-related changes in cholera incidence and several environmental variables including rainfall. El Niño profoundly changed the annual geographic distribution of cholera in Africa from 2000-2014, shifting the burden to continental East Africa, where almost 50,000 additional cases occur during El Niño years. Cholera incidence during El Niño years was higher in regions of East Africa with increased rainfall, but incidence was also higher in some areas with decreased rainfall suggesting a complex relationship between rainfall and cholera incidence. Here we show clear evidence for a shift in the distribution of cholera incidence throughout Africa in El Niño and non-El Niño years, likely mediated by El Niño's impact on local climatic factors. Knowledge of this relationship between cholera and climate patterns coupled with El Niño forecasting could be used to notify countries in Africa when they are likely to see

  11. An important role for adenine, cholera toxin, hydrocortisone and triiodothyronine in the proliferation, self-renewal and differentiation of limbal stem cells in vitro.

    Science.gov (United States)

    Yu, Min; Bojic, Sanja; Figueiredo, Gustavo S; Rooney, Paul; de Havilland, Julian; Dickinson, Anne; Figueiredo, Francisco C; Lako, Majlinda

    2016-11-01

    The cornea is a self-renewing tissue located at the front of the eye. Its transparency is essential for allowing light to focus onto the retina for visual perception. The continuous renewal of corneal epithelium is supported by limbal stem cells (LSCs) which are located in the border region between conjunctiva and cornea known as the limbus. Ex vivo expansion of LSCs has been successfully applied in the last two decades to treat patients with limbal stem cell deficiency (LSCD). Various methods have been used for their expansion, yet the most widely used culture media contains a number of ingredients derived from animal sources which may compromise the safety profile of human LSC transplantation. In this study we sought to understand the role of these components namely adenine, cholera toxin, hydrocortisone and triiodothyronine with the aim of re-defining a safe and GMP compatible minimal media for the ex vivo expansion of LSCs on human amniotic membrane. Our data suggest that all four components play a critical role in maintaining LSC proliferation and promoting LSC self-renewal. However removal of adenine and triiodothyronine had a more profound impact and led to LSC differentiation and loss of viability respectively, suggesting their essential role for ex vivo expansion of LSCs. Replacement of each of the components with GMP-grade reagents resulted in equal growth to non-GMP grade media, however an enhanced differentiation of LSCs was observed, suggesting that additional combinations of GMP grade reagents need to be tested to achieve similar or better level of LSC maintenance in the same manner as the traditional LSC media. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

  12. 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.

  13. Geospatial and temporal patterns of annual cholera outbreaks in Matlab, Bangladesh

    Science.gov (United States)

    Majumder, M. S.; de Klerk, K.; Meyers, D.

    2012-12-01

    Cholera is a waterborne diarrheal disease endemic to Bangladesh, resulting in 1 million diagnoses annually. Such disease burden results in incalculable lost wages and treatment expenses, taken from the pockets of an already impoverished society. Two seasonally correlated outbreaks of cholera occur in Bangladesh every year. In the spring and early summer, the Bay of Bengal - which serves as a natural reservoir for the cholera bacteria - flows inland, causing the first outbreak amongst coastal communities. Waste containing the cholera bacteria enters the sewage system and remains untreated due to poor water and sanitation infrastructure. Therefore, during the following monsoon season, flooding of cholera-contaminated sewage into drinking water sources results in a second outbreak. Though considered common knowledge among local populations, this geographic and temporal progression has not been empirically verified in the current literature. The aim of our ongoing study is to systematically analyze the seasonal trajectory of endemic cholera in Bangladesh. This paper discusses the results obtained from a comprehensive survey of available cholera data from the International Centre of Diarrheal Disease Research, Bangladesh (ICDDR,B) in Matlab, Bangladesh. Matlab thana is a near-coastal community that consists of 142 villages. Monsoon season takes place from June through October. Due to its proximity to the Meghna River, which opens into the Bay of Bengal, the area experiences significant flooding during these months. Using 10 years of geographically referenced cholera data, cases were plotted in time and space. Preliminary patterns suggest that villages closer to the Meghna River experience the majority of the area's cholera outbreaks and that case count is highest in late spring and late fall. April/May and November/December represent 25% and 23% of total annual case counts respectively. Moreover, villages further from the coastline demonstrate 57% higher relative

  14. A novel kit for rapid detection of Vibrio cholerae O1.

    OpenAIRE

    Hasan, J A; Huq, A; Tamplin, M L; Siebeling, R J; Colwell, R R

    1994-01-01

    We report on the development and testing of a novel, rapid, colorimetric immunodiagnostic kit, Cholera SMART, for direct detection of the presence of Vibrio cholerae O1 in clinical specimens. Unlike conventional culture methods requiring several days to complete, the Cholera SMART kit can be used directly in the field by untrained or minimally skilled personnel to detect V. cholerae O1 in less than 15 min, without cumbersome laboratory equipment. A total of 120 clinical and environmental bact...

  15. antimicrobial susceptibility pattern of vibrio cholerae 01 strains

    African Journals Online (AJOL)

    hi-tech

    East African Medical Journal Vol. 77 No. 7 July 2000. ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF VIBRIO CHOLERAE 01 STRAINS DURING TWO CHOLERA OUTBREAKS IN DAR ES SALAAM,. TANZANIA. W.K. Urassa, MD, MSc, MMed, Lecturer, Department of Microbiology and Immunology, Muhimbili University ...

  16. Human Mobility Patterns and Cholera Epidemics: a Spatially Explicit Modeling Approach

    Science.gov (United States)

    Mari, L.; Bertuzzo, E.; Righetto, L.; Casagrandi, R.; Gatto, M.; Rodriguez-Iturbe, I.; Rinaldo, A.

    2010-12-01

    Cholera is an acute enteric disease caused by the ingestion of water or food contaminated by the bacterium Vibrio cholerae. Although most infected individuals do not develop severe symptoms, their stool may contain huge quantities of V.~cholerae cells. Therefore, while traveling or commuting, asymptomatic carriers can be responsible for the long-range dissemination of the disease. As a consequence, human mobility is an alternative and efficient driver for the spread of cholera, whose primary propagation pathway is hydrological transport through river networks. We present a multi-layer network model that accounts for the interplay between epidemiological dynamics, hydrological transport and long-distance dissemination of V.~cholerae due to human movement. In particular, building on top of state-of-the-art spatially explicit models for cholera spread through surface waters, we describe human movement and its effects on the propagation of the disease by means of a gravity-model approach borrowed from transportation theory. Gravity-like contact processes have been widely used in epidemiology, because they can satisfactorily depict human movement when data on actual mobility patterns are not available. We test our model against epidemiological data recorded during the cholera outbreak occurred in the KwaZulu-Natal province of South Africa during years 2000--2001. We show that human mobility does actually play an important role in the formation of the spatiotemporal patterns of cholera epidemics. In particular, long-range human movement may determine inter-catchment dissemination of V.~cholerae cells, thus in turn explaining the emergence of epidemic patterns that cannot be produced by hydrological transport alone. We also show that particular attention has to be devoted to study how heterogeneously distributed drinking water supplies and sanitation conditions may affect cholera transmission.

  17. Epidemic waves of cholera in the last two decades in Mozambique.

    Science.gov (United States)

    Langa, José Paulo; Sema, Cynthia; De Deus, Nilsa; Colombo, Mauro Maria; Taviani, Elisa

    2015-07-04

    Africa is increasingly affected by cholera. In Mozambique, cholera appeared in the early 1970s when the seventh pandemic entered Africa from the Indian subcontinent. In the following decades, several epidemics were registered in the country, the 1997-1999 epidemic being the most extended. Since then, Mozambique has been considered an endemic area for cholera, characterized by yearly outbreaks occurring with a seasonal pattern. At least three pandemic variants are thought to have originated in the Indian subcontinent and spread worldwide at different times. To understand the epidemiology of cholera in Mozambique, whether the disease re-emerges periodically or is imported by different routes of transmission, we investigated clinical V. cholerae O1 isolated during 1997-1999 and 2012-2014 epidemics. By detecting and characterizing seven genetic elements, the mobilome profile of each isolate was obtained. By comparing it to known seventh pandemic reference strains, it was possible to discern among different V. cholerae O1 variants active in the country. During 1997-1999, epidemic strains showed two different genetic profiles, both related to a pandemic clone that originated from India and was reported in other African countries in the 1990s. Isolates from 2012-2014 outbreaks showed a genetic background related to the pandemic strains currently active as the prevalent causative agent of cholera worldwide. Despite cholera being endemic in Mozambique, the epidemiology of the disease in the past 20 years has been strongly influenced by the cholera seventh pandemic waves that originated in the Indian subcontinent.

  18. c-Jun Proto-Oncoprotein Plays a Protective Role in Lung Epithelial Cells Exposed to Staphylococcal α-Toxin

    Directory of Open Access Journals (Sweden)

    Alejandro J. Moyano

    2018-05-01

    Full Text Available c-Jun is a member of the early mammalian transcriptional regulators belonging to the AP-1 family, which participates in a wide range of cellular processes such as proliferation, apoptosis, tumorigenesis, and differentiation. Despite its established role in cell survival upon stress, its participation in the stress response induced by bacterial infections has been poorly investigated. To study the potential role of c-Jun in this context we choose the widely studied α-toxin produced by Staphylococcus aureus, a pore-forming toxin that is a critical virulence factor in the pathogenesis of these bacteria. We analyzed the effect of α-toxin treatment in the activation, expression, and protein levels of c-Jun in A549 lung epithelial cells. Furthermore, we explored the role of c-Jun in the cellular fate after exposure to α-toxin. Our results show that staphylococcal α-toxin per se is able to activate c-Jun by inducing phosphorylation of its Serine 73 residue. Silencing of the JNK (c-Jun N-terminal Kinase signaling pathway abrogated most of this activation. On the contrary, silencing of the ERK (Extracellular Signal-Regulated Kinase pathway exacerbated this response. Intriguingly, while the exposure to α-toxin induced a marked increase in the levels of c-Jun transcripts, c-Jun protein levels noticeably decreased in the same time-frame as a consequence of active proteolytic degradation through the proteasome-dependent pathway. In addition, we established that c-Jun promoted cell survival when cells were challenged with α-toxin. Similarly, c-Jun phosphorylation was also induced in cells upon intoxication with the cytolysin produced by Vibrio cholerae in a JNK-dependent manner, suggesting that c-Jun-JNK axis would be a conserved responsive cellular pathway to pore-forming toxins. This study contributes to understanding the role of the multifaceted c-Jun proto-oncoprotein in cell response to bacterial pore-forming toxins, positioning it as a relevant

  19. Potential impact of reactive vaccination in controlling cholera ...

    African Journals Online (AJOL)

    Background. To contain ongoing cholera outbreaks, the World Health Organization has suggested that reactive vaccination should be considered in addition to its previous control measures. Objectives. To explore the potential impact of a hypothetical reactive oral cholera vaccination using the example of the recent ...

  20. Effectiveness of Oral Cholera Vaccine in Haiti: 37-Month Follow-Up.

    Science.gov (United States)

    Sévère, Karine; Rouzier, Vanessa; Anglade, Stravinsky Benedict; Bertil, Claudin; Joseph, Patrice; Deroncelay, Alexandra; Mabou, Marie Marcelle; Wright, Peter F; Guillaume, Florence Duperval; Pape, Jean William

    2016-05-04

    The first oral cholera vaccine (OCV) campaign, since its prequalification by the World Health Organization, in response to an ongoing cholera epidemic (reactive vaccination) was successfully conducted in a poor urban slum of approximately 70,000 inhabitants in Port-au-Prince, Haiti, in 2012. Vaccine coverage was 75% of the target population. This report documents the impact of OCV in reducing the number of culture-confirmed cases of cholera admitted to the Groupe Haïtien d'Etude du Sarcome de Kaposi et des Infections Opportunistes (GHESKIO) cholera treatment center from that community in the 37 months postvaccination (April 2012-April 30, 2015). Of 1,788 patients with culture-confirmed cholera, 1,770 (99%) were either from outside the vaccine area (1,400 cases) or from the vaccinated community who had not received OCV (370 cases). Of the 388 people from the catchment area who developed culture-confirmed cholera, 370 occurred among the 17,643 people who had not been vaccinated (2.1%) and the remaining 18 occurred among the 52,357 people (0.034%) who had been vaccinated (P cholera in outbreak settings. © The American Society of Tropical Medicine and Hygiene.

  1. The Orphans of Cholera Morbus in Yucatan, 1833

    Directory of Open Access Journals (Sweden)

    Elsa Malvido

    2013-07-01

    Full Text Available This essay discusses the phenomenon of orphanhood which affected a large number of children after the cholera epidemic that struck Yucatan in July 1833. Moreover, it inquires into the fate of children whose parents died of cholera, the role played by kinship networks to provide them with shelter, and the influence of the Church and the State on the situation. Based on first hand sources, the author suggests that the orphanhood produced by cholera served as a pretext for economically and socially privileged groups to get hold of free labor force both for domestic service and hacienda work.

  2. Oral cholera vaccine--for whom, when, and why?

    Science.gov (United States)

    Topps, Maureen H

    2006-01-01

    The search for a safe, effective, well tolerated, low cost vaccine against the ancient cholera enemy has been ongoing since the 19th century and has been revitalized in the past two decades since the advent of recombinant technology. Large-scale field trials have readily demonstrated the tolerability and safety of oral cholera vaccine in various forms. Variable levels of protection have been shown and one challenge has been to demonstrate whether this is a cost effective treatment in differing environments including its use in endemic and epidemic areas as well as for travelers. A review of recent literature was undertaken to assess the effectiveness and uses of currently available oral cholera vaccine. While the evidence does not support the creation of formal guidelines, some clear recommendations can be made. There is undoubtedly the potential to reduce the burden of illness both in endemic and epidemic situations. For travelers, certain higher risk groups may benefit from protection against cholera. More significantly, the short term cross-protection afforded by whole cell, B subunit (WC BS) oral cholera vaccine formulations against enterotoxigenic E. coli, (ETEC), the commonest causative agent of traveler's diarrhoea, may prove to be the most important raison d'être.

  3. Cholera in the United States, 1965-1991. Risks at home and abroad.

    Science.gov (United States)

    Weber, J T; Levine, W C; Hopkins, D P; Tauxe, R V

    1994-03-14

    To assess risks for cholera in the United States. Review of published reports of cholera outbreaks and sporadic cases and Centers for Disease Control and Prevention (CDC) memoranda and laboratory reports. Persons with symptomatic laboratory-diagnosed cholera treated in the United States and territories. From 1965 through 1991, 136 cases of cholera were reported. Fifty-three percent of the patients were hospitalized and three persons died (case-fatality rate, 0.02). Ninety-three infections were acquired in the United States and 42 overseas; for one case the source was unknown. Domestically acquired cholera was largely related to the endemic Gulf Coast focus of Vibrio cholerae 01 (56 cases). The major domestic food vehicle was shellfish, particularly crabs harvested from the Gulf of Mexico or nearby estuaries. In 1991, 14 (54%) of 26 domestically acquired cases were caused by food from Ecuador (n = 11) and Thailand (n = 3). During 1991, the first cases of cholera in travelers returning from South America were reported. In 1991, the rate of cholera among air travelers returning from South America was estimated as 0.3 per 100,000; among air travelers returning from Ecuador, 2.6 per 100,000. Cholera remains a small but persistent risk in the United States and for travelers. An endemic focus on the Gulf Coast, the continuing global pandemic, and the epidemic in South America make this likely to continue for years to come. Physicians should know how to diagnose and treat cholera and should report all suspected cases to their state health departments.

  4. Comparing sociocultural features of cholera in three endemic African settings

    Science.gov (United States)

    2013-01-01

    Background Cholera mainly affects developing countries where safe water supply and sanitation infrastructure are often rudimentary. Sub-Saharan Africa is a cholera hotspot. Effective cholera control requires not only a professional assessment, but also consideration of community-based priorities. The present work compares local sociocultural features of endemic cholera in urban and rural sites from three field studies in southeastern Democratic Republic of Congo (SE-DRC), western Kenya and Zanzibar. Methods A vignette-based semistructured interview was used in 2008 in Zanzibar to study sociocultural features of cholera-related illness among 356 men and women from urban and rural communities. Similar cross-sectional surveys were performed in western Kenya (n = 379) and in SE-DRC (n = 360) in 2010. Systematic comparison across all settings considered the following domains: illness identification; perceived seriousness, potential fatality and past household episodes; illness-related experience; meaning; knowledge of prevention; help-seeking behavior; and perceived vulnerability. Results Cholera is well known in all three settings and is understood to have a significant impact on people’s lives. Its social impact was mainly characterized by financial concerns. Problems with unsafe water, sanitation and dirty environments were the most common perceived causes across settings; nonetheless, non-biomedical explanations were widespread in rural areas of SE-DRC and Zanzibar. Safe food and water and vaccines were prioritized for prevention in SE-DRC. Safe water was prioritized in western Kenya along with sanitation and health education. The latter two were also prioritized in Zanzibar. Use of oral rehydration solutions and rehydration was a top priority everywhere; healthcare facilities were universally reported as a primary source of help. Respondents in SE-DRC and Zanzibar reported cholera as affecting almost everybody without differentiating much for gender, age

  5. Dealing with cholera: exclusively the domain of environmental ...

    African Journals Online (AJOL)

    Cholera outbreaks have a profound impact on the health and well-being of communities. Especially young children are vulnerable to the disease and schools report high absenteeism during epidemics. There is both the perception and evidence thereof, that educating communities about cholera (its prevention and ...

  6. Epidemic Cholera and American Reform Movements in the 19th Century

    Directory of Open Access Journals (Sweden)

    Seohyung KIM

    2015-12-01

    Full Text Available The 19th century was the age of great reform in American history. After constructing of the canal and railroads, the industrialization began and American society changed so rapidly. In this period, there were so many social crisis and American people tried to solve these problems within the several reform movements. These reform movements were the driving forces to control cholera during the 19th century. Cholera was the endemic disease in Bengal, India, but after the 19th century it had spread globally by the development of trade networks. The 1832 cholera in the United States was the first epidemic cholera in American history. The mortality of cholera was so high, but it was very hard to find out the cause of this fatal infectious disease. So, different social discourses happened to control epidemic cholera in the 19th century, these can be understood within the similar context of American reform movements during this period. Board of Health in New York States made a new public health act to control cholera in 1832, it was ineffective. Some people insisted that the cause of this infectious disease was the corruption of the United States. They emphasized unjust and immoral system in American society. Moral reform expanded to Nativism, because lots of Irish immigrants were the victims of cholera. So, epidemic cholera was the opportunity to spread the desire for moral reform. To control cholera in 1849, the sanitary reform in Britain had affected. The fact that it was so important to improve and maintain the water quality for the control and prevention of disease spread, the sanitary reform happened. There were two different sphere of the sanitary reform. The former was the private reform to improve sewer or privy, the latter was the public reform to build sewage facilities. The 1849 cholera had an important meaning, because the social discourse, which had emphasized the sanitation of people or home expanded to the public sphere. When cholera

  7. [Epidemic Cholera and American Reform Movements in the 19th Century].

    Science.gov (United States)

    Kim, Seohyung

    2015-12-01

    The 19th century was the age of great reform in American history. After constructing of the canal and railroads, the industrialization began and American society changed so rapidly. In this period, there were so many social crisis and American people tried to solve these problems within the several reform movements. These reform movements were the driving forces to control cholera during the 19th century. Cholera was the endemic disease in Bengal, India, but after the 19th century it had spread globally by the development of trade networks. The 1832 cholera in the United States was the first epidemic cholera in American history. The mortality of cholera was so high, but it was very hard to find out the cause of this fatal infectious disease. So, different social discourses happened to control epidemic cholera in the 19th century, these can be understood within the similar context of American reform movements during this period. Board of Health in New York States made a new public health act to control cholera in 1832, it was ineffective. Some people insisted that the cause of this infectious disease was the corruption of the United States. They emphasized unjust and immoral system in American society. Moral reform expanded to Nativism, because lots of Irish immigrants were the victims of cholera. So, epidemic cholera was the opportunity to spread the desire for moral reform. To control cholera in 1849, the sanitary reform in Britain had affected. The fact that it was so important to improve and maintain the water quality for the control and prevention of disease spread, the sanitary reform happened. There were two different sphere of the sanitary reform. The former was the private reform to improve sewer or privy, the latter was the public reform to build sewage facilities. The 1849 cholera had an important meaning, because the social discourse, which had emphasized the sanitation of people or home expanded to the public sphere. When cholera broke out in 1866 again

  8. Knowledge of, attitudes toward, and preventive practices relating to cholera and oral cholera vaccine among urban high-risk groups: findings of a cross-sectional study in Dhaka, Bangladesh

    OpenAIRE

    Wahed, Tasnuva; Kaukab, Sheikh Shah Tanvir; Saha, Nirod Chandra; Khan, Iqbal Ansary; Khanam, Farhana; Chowdhury, Fahima; Saha, Amit; Khan, Ashraful Islam; Siddik, Ashraf Uddin; Cravioto, Alejandro; Qadri, Firdausi; Uddin, Jasim

    2013-01-01

    Background In endemic countries such as Bangladesh, consequences of cholera place an enormous financial and social burden on patients and their families. Cholera vaccines not only provide health benefits to susceptible populations but also have effects on the earning capabilities and financial stability of the family. Community-based research and evaluations are necessary to understand perceptions about and practices of the community relating to cholera and oral cholera vaccines. This may hel...

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

    International Nuclear Information System (INIS)

    Naseer, M.; Jamali, T.

    2014-01-01

    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. (author)

  10. Epidemic Cholera in a Crowded Urban Environment, Port-au-Prince, Haiti

    OpenAIRE

    Dunkle, Stacie E.; Mba-Jonas, Adamma; Loharikar, Anagha; Fouché, Bernadette; Peck, Mireille; Ayers, Tracy; Archer, W. Roodly; De Rochars, Valery M. Beau; Bender, Thomas; Moffett, Daphne B.; Tappero, Jordan W.; Dahourou, George; Roels, Thierry H.; Quick, Robert

    2011-01-01

    We conducted a case–control study to investigate factors associated with epidemic cholera. Water treatment and handwashing may have been protective, highlighting the need for personal hygiene for cholera prevention in contaminated urban environments. We also found a diverse diet, a possible proxy for improved nutrition, was protective against cholera.

  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. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  12. Protection against cholera from killed whole-cell oral cholera vaccines: a systematic review and meta-analysis.

    Science.gov (United States)

    Bi, Qifang; Ferreras, Eva; Pezzoli, Lorenzo; Legros, Dominique; Ivers, Louise C; Date, Kashmira; Qadri, Firdausi; Digilio, Laura; Sack, David A; Ali, Mohammad; Lessler, Justin; Luquero, Francisco J; Azman, Andrew S

    2017-10-01

    Killed whole-cell oral cholera vaccines (kOCVs) are becoming a standard cholera control and prevention tool. However, vaccine efficacy and direct effectiveness estimates have varied, with differences in study design, location, follow-up duration, and vaccine composition posing challenges for public health decision making. We did a systematic review and meta-analysis to generate average estimates of kOCV efficacy and direct effectiveness from the available literature. For this systematic review and meta-analysis, we searched PubMed, Embase, Scopus, and the Cochrane Review Library on July 9, 2016, and ISI Web of Science on July 11, 2016, for randomised controlled trials and observational studies that reported estimates of direct protection against medically attended confirmed cholera conferred by kOCVs. We included studies published on any date in English, Spanish, French, or Chinese. We extracted from the published reports the primary efficacy and effectiveness estimates from each study and also estimates according to number of vaccine doses, duration, and age group. The main study outcome was average efficacy and direct effectiveness of two kOCV doses, which we estimated with random-effect models. This study is registered with PROSPERO, number CRD42016048232. Seven trials (with 695 patients with cholera) and six observational studies (217 patients with cholera) met the inclusion criteria, with an average two-dose efficacy of 58% (95% CI 42-69, I 2 =58%) and effectiveness of 76% (62-85, I 2 =0). Average two-dose efficacy in children younger than 5 years (30% [95% CI 15-42], I 2 =0%) was lower than in those 5 years or older (64% [58-70], I 2 =0%; pcholera for at least 3 years. One kOCV dose provides at least short-term protection, which has important implications for outbreak management. kOCVs are effective tools for cholera control. The Bill & Melinda Gates Foundation. Copyright This is an Open Access article published under the CC BY 3.0 IGO license which permits

  13. Cholera in Children

    Science.gov (United States)

    ... oral antibiotic will be given. More serious skin infections and bacteria in the blood are treated in the hospital with intravenous antibiotics. In some cases, your child may require surgery to drain pus and damaged tissues. Prevention V cholerae can be killed by boiling, filtering, ...

  14. Cholera Incidence and Mortality in Sub-Saharan African Sites during Multi-country Surveillance.

    Science.gov (United States)

    Sauvageot, Delphine; Njanpop-Lafourcade, Berthe-Marie; Akilimali, Laurent; Anne, Jean-Claude; Bidjada, Pawou; Bompangue, Didier; Bwire, Godfrey; Coulibaly, Daouda; Dengo-Baloi, Liliana; Dosso, Mireille; Orach, Christopher Garimoi; Inguane, Dorteia; Kagirita, Atek; Kacou-N'Douba, Adele; Keita, Sakoba; Kere Banla, Abiba; Kouame, Yao Jean-Pierre; Landoh, Dadja Essoya; Langa, Jose Paulo; Makumbi, Issa; Miwanda, Berthe; Malimbo, Muggaga; Mutombo, Guy; Mutombo, Annie; NGuetta, Emilienne Niamke; Saliou, Mamadou; Sarr, Veronique; Senga, Raphael Kakongo; Sory, Fode; Sema, Cynthia; Tante, Ouyi Valentin; Gessner, Bradford D; Mengel, Martin A

    2016-05-01

    Cholera burden in Africa remains unknown, often because of weak national surveillance systems. We analyzed data from the African Cholera Surveillance Network (www.africhol.org). During June 2011-December 2013, we conducted enhanced surveillance in seven zones and four outbreak sites in Togo, the Democratic Republic of Congo (DRC), Guinea, Uganda, Mozambique and Cote d'Ivoire. All health facilities treating cholera cases were included. Cholera incidences were calculated using culture-confirmed cholera cases and culture-confirmed cholera cases corrected for lack of culture testing usually due to overwhelmed health systems and imperfect test sensitivity. Of 13,377 reported suspected cases, 34% occurred in Conakry, Guinea, 47% in Goma, DRC, and 19% in the remaining sites. From 0-40% of suspected cases were aged under five years and from 0.3-86% had rice water stools. Within surveillance zones, 0-37% of suspected cases had confirmed cholera compared to 27-38% during outbreaks. Annual confirmed incidence per 10,000 population was cholera incidence, age distribution, clinical presentation, culture confirmation, and testing frequency. These results can help guide preventive activities, including vaccine use.

  15. Toxigenic Vibrio cholerae O1 in vegetables and fish raised in wastewater irrigated fields and stabilization ponds during a non-cholera outbreak period in Morogoro, Tanzania

    DEFF Research Database (Denmark)

    Hounmanou, Yaovi M G; Mdegela, Robinson H; Dougnon, Tamègnon V

    2016-01-01

    gene (tcpA) and the haemolysin gene (hlyA). RESULTS: The prevalence of V. cholerae in wastewater, vegetables and fish was 36.7, 21.7 and 23.3 %, respectively. Two isolates from fish gills were V. cholerae O1 and tested positive for ctx and tcpA. One of these contained in addition the hlyA gene while......BACKGROUND: Cholera, one of the world's deadliest infectious diseases, remains rampant and frequent in Tanzania and thus hinders existing control measures. The present study was undertaken to evaluate the occurrence of toxigenic Vibrio cholerae O1 in wastewater, fish and vegetables during a non......-outbreak period in Morogoro, Tanzania. METHODS: From October 2014 to February 2015, 60 wastewater samples, 60 fish samples from sewage stabilization ponds and 60 wastewater irrigated vegetable samples were collected. Samples were cultured for identification of V. cholerae using conventional bacteriological...

  16. Prevalence of Vibrio cholerae O1 serogroup in Assam, India: A hospital-based study.

    Science.gov (United States)

    Sharma, Ajanta; Dutta, Bornali Sarmah; Rasul, Elmy Samsun; Barkataki, Dipa; Saikia, Anjanamoyee; Hazarika, Naba Kumar

    2017-09-01

    Although cholera remains to be an important public health problem, studies on reliable population-based estimates of laboratory confirmed cholera in endemic areas are limited worldwide. The aim of this hospital-based study was to evaluate the prevalence of Vibrio cholerae serogroup in Assam, India, during 2003-2013. Stool samples/rectal swabs were collected from acute watery diarrhoea (AWD) cases during 2003-2013 and processed by standard microbiological procedures. Antibiotic sensitivity test was done following the Clinical and Laboratory Standards Institute guidelines. Year-wise epidemiological trend of cholera was analyzed. Cholera contributed to 3.93 per cent of AWD cases. In Assam, cholera was found to be more prevalent in the rural areas (6.7%) followed by the tea gardens (5.06%), urban slum (1.9%) and urban areas (1.4%). Highest proportion of cholera (13.7%) was observed in 0-10 yr age group. Of them, 11.5 per cent belonged to 0-5 yr age group. V. cholerae O1 El Tor serotype Ogawa was the predominant isolate. Multiple drug-resistant isolates of V. cholerae O1 Ogawa were reported in the study. Emergence of resistance amongst V. cholerae towards many antibiotics is a matter of concern. Hence, continuous surveillance for diarrhoeal disorders is necessary to control the future outbreaks of cholera in this region.

  17. Endoplasmic Reticulum-Targeted Subunit Toxins Provide a New Approach to Rescue Misfolded Mutant Proteins and Revert Cell Models of Genetic Diseases.

    Directory of Open Access Journals (Sweden)

    Humaira Adnan

    Full Text Available Many germ line diseases stem from a relatively minor disturbance in mutant protein endoplasmic reticulum (ER 3D assembly. Chaperones are recruited which, on failure to correct folding, sort the mutant for retrotranslocation and cytosolic proteasomal degradation (ER-associated degradation-ERAD, to initiate/exacerbate deficiency-disease symptoms. Several bacterial (and plant subunit toxins, retrograde transport to the ER after initial cell surface receptor binding/internalization. The A subunit has evolved to mimic a misfolded protein and hijack the ERAD membrane translocon (dislocon, to effect cytosolic access and cytopathology. We show such toxins compete for ERAD to rescue endogenous misfolded proteins. Cholera toxin or verotoxin (Shiga toxin containing genetically inactivated (± an N-terminal polyleucine tail A subunit can, within 2-4 hrs, temporarily increase F508delCFTR protein, the major cystic fibrosis (CF mutant (5-10x, F508delCFTR Golgi maturation (<10x, cell surface expression (20x and chloride transport (2x in F508del CFTR transfected cells and patient-derived F508delCFTR bronchiolar epithelia, without apparent cytopathology. These toxoids also increase glucocerobrosidase (GCC in N370SGCC Gaucher Disease fibroblasts (3x, another ERAD-exacerbated misfiling disease. We identify a new, potentially benign approach to the treatment of certain genetic protein misfolding diseases.

  18. Cholera in Thomas Mann's Death in Venice.

    Science.gov (United States)

    Rütten, Thomas

    2009-01-01

    The article sets the cholera motif in Thomas Mann's famous novella Death in Venice against the historical context from which it partially originates. It is shown that this motif, while undoubtedly appropriated to serve Mann's own poetic ends, has a solid grounding in historical and autobiographical fact, thus blurring the boundaries between fact and fiction. The article illustrates the verifiable events of the outbreak of the Venetian cholera epidemic in May 1911, which Mann partly witnessed himself, during a holiday trip to Brioni and Venice, and partly heard and read about. It is established that Thomas Mann's account of the cholera in Venice in his novella is characterised by a rare and almost preternatural insightfulness into an otherwise murky affair that was marked by rumours, speculations and denials.

  19. Modelling cholera epidemics: the role of waterways, human mobility and sanitation.

    Science.gov (United States)

    Mari, L; Bertuzzo, E; Righetto, L; Casagrandi, R; Gatto, M; Rodriguez-Iturbe, I; Rinaldo, A

    2012-02-07

    We investigate the role of human mobility as a driver for long-range spreading of cholera infections, which primarily propagate through hydrologically controlled ecological corridors. Our aim is to build a spatially explicit model of a disease epidemic, which is relevant to both social and scientific issues. We present a two-layer network model that accounts for the interplay between epidemiological dynamics, hydrological transport and long-distance dissemination of the pathogen Vibrio cholerae owing to host movement, described here by means of a gravity-model approach. We test our model against epidemiological data recorded during the extensive cholera outbreak occurred in the KwaZulu-Natal province of South Africa during 2000-2001. We show that long-range human movement is fundamental in quantifying otherwise unexplained inter-catchment transport of V. cholerae, thus playing a key role in the formation of regional patterns of cholera epidemics. We also show quantitatively how heterogeneously distributed drinking water supplies and sanitation conditions may affect large-scale cholera transmission, and analyse the effects of different sanitation policies.

  20. Modelling cholera epidemics: the role of waterways, human mobility and sanitation

    Science.gov (United States)

    Mari, L.; Bertuzzo, E.; Righetto, L.; Casagrandi, R.; Gatto, M.; Rodriguez-Iturbe, I.; Rinaldo, A.

    2012-01-01

    We investigate the role of human mobility as a driver for long-range spreading of cholera infections, which primarily propagate through hydrologically controlled ecological corridors. Our aim is to build a spatially explicit model of a disease epidemic, which is relevant to both social and scientific issues. We present a two-layer network model that accounts for the interplay between epidemiological dynamics, hydrological transport and long-distance dissemination of the pathogen Vibrio cholerae owing to host movement, described here by means of a gravity-model approach. We test our model against epidemiological data recorded during the extensive cholera outbreak occurred in the KwaZulu-Natal province of South Africa during 2000–2001. We show that long-range human movement is fundamental in quantifying otherwise unexplained inter-catchment transport of V. cholerae, thus playing a key role in the formation of regional patterns of cholera epidemics. We also show quantitatively how heterogeneously distributed drinking water supplies and sanitation conditions may affect large-scale cholera transmission, and analyse the effects of different sanitation policies. PMID:21752809

  1. Cholera in Haiti: Reproductive numbers and vaccination coverage estimates

    Science.gov (United States)

    Mukandavire, Zindoga; Smith, David L.; Morris, J. Glenn, Jr.

    2013-01-01

    Cholera reappeared in Haiti in October, 2010 after decades of absence. Cases were first detected in Artibonite region and in the ensuing months the disease spread to every department in the country. The rate of increase in the number of cases at the start of epidemics provides valuable information about the basic reproductive number (). Quantitative analysis of such data gives useful information for planning and evaluating disease control interventions, including vaccination. Using a mathematical model, we fitted data on the cumulative number of reported hospitalized cholera cases in Haiti. varied by department, ranging from 1.06 to 2.63. At a national level, 46% vaccination coverage would result in an () cholera vaccines in endemic and non-endemic regions, our results suggest that moderate cholera vaccine coverage would be an important element of disease control in Haiti.

  2. Antimicrobial Resistance Risks of Cholera Prophylaxis for United Nations Peacekeepers.

    Science.gov (United States)

    Kunkel, Amber; Lewnard, Joseph A; Pitzer, Virginia E; Cohen, Ted

    2017-08-01

    More than 5 years after a United Nations peacekeeping battalion introduced cholera to Haiti, over 150,000 peacekeepers continue to be deployed annually from countries where cholera is endemic. The United Nations has thus far declined to provide antimicrobial chemoprophylaxis to peacekeepers, a policy based largely on concerns that the risks of drug resistance generation and spread would outweigh the potential benefits of preventing future cholera importations. In this study, we sought to better understand the relative benefits and risks of cholera chemoprophylaxis for peacekeepers in terms of antibiotic resistance. Using a stochastic model to quantify the potential impact of chemoprophylaxis on importation and transmission of drug-resistant and drug-sensitive Vibrio cholerae , we found that chemoprophylaxis would decrease the probability of cholera importation but would increase the expected number of drug-resistant infections if an importation event were to occur. Despite this potential increase, we found that at least 10 drug-sensitive infections would likely be averted per excess drug-resistant infection under a wide range of assumptions about the underlying prevalence of drug resistance and risk of acquired resistance. Given these findings, policymakers should reconsider whether the potential resistance risks of providing antimicrobial chemoprophylaxis to peacekeepers are sufficient to outweigh the anticipated benefits. Copyright © 2017 American Society for Microbiology.

  3. Satellite Based Assessment of Hydroclimatic Conditions Related to Cholera in Zimbabwe.

    Directory of Open Access Journals (Sweden)

    Antarpreet Jutla

    Full Text Available Cholera, an infectious diarrheal disease, has been shown to be associated with large scale hydroclimatic processes. The sudden and sporadic occurrence of epidemic cholera is linked with high mortality rates, in part, due to uncertainty in timing and location of outbreaks. Improved understanding of the relationship between pathogenic abundance and climatic processes allows prediction of disease outbreak to be an achievable goal. In this study, we show association of large scale hydroclimatic processes with the cholera epidemic in Zimbabwe reported to have begun in Chitungwiza, a city in Mashonaland East province, in August, 2008.Climatic factors in the region were found to be associated with triggering cholera outbreak and are shown to be related to anomalies of temperature and precipitation, validating the hypothesis that poor conditions of sanitation, coupled with elevated temperatures, and followed by heavy rainfall can initiate outbreaks of cholera. Spatial estimation by satellite of precipitation and global gridded air temperature captured sensitivities in hydroclimatic conditions that permitted identification of the location in the region where the disease outbreak began.Satellite derived hydroclimatic processes can be used to capture environmental conditions related to epidemic cholera, as occurred in Zimbabwe, thereby providing an early warning system. Since cholera cannot be eradicated because the causative agent, Vibrio cholerae, is autochthonous to the aquatic environment, prediction of conditions favorable for its growth and estimation of risks of triggering the disease in a given population can be used to alert responders, potentially decreasing infection and saving lives.

  4. Evaluation of Knowledge and Practices Regarding Cholera, Water Treatment, Hygiene, and Sanitation Before and After an Oral Cholera Vaccination Campaign-Haiti, 2013-2014.

    Science.gov (United States)

    Childs, Lana; François, Jeannot; Choudhury, Alina; Wannemuehler, Kathleen; Dismer, Amber; Hyde, Terri B; Yen, Catherine Y; Date, Kashmira A; Juin, Stanley; Katz, Mark A; Kantor, Erica Felker; Routh, Janell; Etheart, Melissa; Wright, Tracie; Adrien, Paul; Tohme, Rania A

    2016-12-07

    In 2013, the Government of Haiti implemented its first oral cholera vaccine (OCV) campaign in Petite Anse, an urban setting, and Cerca Carvajal, a rural commune. We conducted and compared responses to two independent cross-sectional knowledge and practices household surveys pre- (N = 297) and post- (N = 302) OCV campaign in Petite Anse. No significant differences in knowledge about causes, symptoms, and prevention of cholera were noted. Compared with precampaign respondents, fewer postcampaign respondents reported treating (66% versus 27%, P treatment practices necessary for cholera and other diarrheal diseases prevention were noted in the postcampaign survey. Future OCV campaigns in Haiti should be used as an opportunity to emphasize the importance of maintaining good water, sanitation, and hygiene practices, and include a comprehensive, integrated approach for cholera control. © The American Society of Tropical Medicine and Hygiene.

  5. [Cholera in 1831. Challenges for science and the federal government].

    Science.gov (United States)

    Stamm-Kuhlmann, T

    1989-01-01

    The peak of the first great cholera pandemic in 1831 fomented the controversy among contagionists and non-contagionists. In the following year the public debate centered around the correct interpretation of the recent experiences with cholera. The central government of the bureaucratic-absolutist monarchy in Prussia adhered to a firmly contagionist interpretation of the disease and reacted accordingly. Local authorities in Königsberg and Berlin and the bourgeoisie in the merchant city of Danzig, however, stressed the destructive consequences of the cordon system. They considered the results of an interruption in trade and industry to be worse than the damage inflicted by the epidemic. The summer of 1831 demonstrated that cholera could not be stopped by the cordons, but the King's medical advisors nevertheless remained contagionists. Non-contagionists put forward several hypotheses to explain the origin and the spreading of cholera, mainly "miasma" theory and the Hippocratic paradigm of "epidemic constitution". The correlation between poverty and disease, however, was widely noticed. Physicians in the city of Bremen pointed to the necessity of sanitary precautions to be taken in cholera-free periods. On the other hand, many "honest" citizens believed that individuals with a "dissolute" conduct of life were more at risk to contract cholera than others. Instead of costly sanitary policies, the well-to-do classes preferred to identify the defense against cholera with the segregation of unwelcome elements of society. The article is based on hitherto unpublished sources from the former Prussian State Archives at Merseburg, GDR, and the State Archive of the Hanseatic City of Bremen.

  6. Identification of burden hotspots and risk factors for cholera in India: An observational study.

    Science.gov (United States)

    Ali, Mohammad; Sen Gupta, Sanjukta; Arora, Nisha; Khasnobis, Pradeep; Venkatesh, Srinivas; Sur, Dipika; Nair, Gopinath B; Sack, David A; Ganguly, Nirmal K

    2017-01-01

    Even though cholera has existed for centuries and many parts of the country have sporadic, endemic and epidemic cholera, it is still an under-recognized health problem in India. A Cholera Expert Group in the country was established to gather evidence and to prepare a road map for control of cholera in India. This paper identifies cholera burden hotspots and factors associated with an increased risk of the disease. We acquired district level data on cholera case reports of 2010-2015 from the Integrated Disease Surveillance Program. Socioeconomic characteristics and coverage of water and sanitation was obtained from the 2011 census. Spatial analysis was performed to identify cholera hotspots, and a zero-inflated Poisson regression was employed to identify the factors associated with cholera and predicted case count in the district. 27,615 cholera cases were reported during the 6-year period. Twenty-four of 36 states of India reported cholera during these years, and 13 states were classified as endemic. Of 641 districts, 78 districts in 15 states were identified as "hotspots" based on the reported cases. On the other hand, 111 districts in nine states were identified as "hotspots" from model-based predicted number of cases. The risk for cholera in a district was negatively associated with the coverage of literate persons, households using treated water source and owning mobile telephone, and positively associated with the coverage of poor sanitation and drainage conditions and urbanization level in the district. The study reaffirms that cholera continues to occur throughout a large part of India and identifies the burden hotspots and risk factors. Policymakers may use the findings of the article to develop a roadmap for prevention and control of cholera in India.

  7. An Optimal Cost Effectiveness Study on Zimbabwe Cholera Seasonal Data from 2008–2011

    Science.gov (United States)

    Sardar, Tridip; Mukhopadhyay, Soumalya; Bhowmick, Amiya Ranjan; Chattopadhyay, Joydev

    2013-01-01

    Incidence of cholera outbreak is a serious issue in underdeveloped and developing countries. In Zimbabwe, after the massive outbreak in 2008–09, cholera cases and deaths are reported every year from some provinces. Substantial number of reported cholera cases in some provinces during and after the epidemic in 2008–09 indicates a plausible presence of seasonality in cholera incidence in those regions. We formulate a compartmental mathematical model with periodic slow-fast transmission rate to study such recurrent occurrences and fitted the model to cumulative cholera cases and deaths for different provinces of Zimbabwe from the beginning of cholera outbreak in 2008–09 to June 2011. Daily and weekly reported cholera incidence data were collected from Zimbabwe epidemiological bulletin, Zimbabwe Daily cholera updates and Office for the Coordination of Humanitarian Affairs Zimbabwe (OCHA, Zimbabwe). For each province, the basic reproduction number () in periodic environment is estimated. To the best of our knowledge, this is probably a pioneering attempt to estimate in periodic environment using real-life data set of cholera epidemic for Zimbabwe. Our estimates of agree with the previous estimate for some provinces but differ significantly for Bulawayo, Mashonaland West, Manicaland, Matabeleland South and Matabeleland North. Seasonal trend in cholera incidence is observed in Harare, Mashonaland West, Mashonaland East, Manicaland and Matabeleland South. Our result suggests that, slow transmission is a dominating factor for cholera transmission in most of these provinces. Our model projects cholera cases and cholera deaths during the end of the epidemic in 2008–09 to January 1, 2012. We also determine an optimal cost-effective control strategy among the four government undertaken interventions namely promoting hand-hygiene & clean water distribution, vaccination, treatment and sanitation for each province. PMID:24312540

  8. Effect of toxin-g from Tityus serrulatus scorpion venom on gastric emptying in rats

    Directory of Open Access Journals (Sweden)

    F. Bucaretchi

    1999-04-01

    Full Text Available The effect of toxin-g from Tityus serrulatus scorpion venom on the gastric emptying of liquids was studied in 176 young adult male Wistar rats (2-3 months of age divided into subgroups of 8 animals each. Toxin-g was injected iv at doses of 25, 37.5, 50 or 100 µg/kg and the effect on gastric emptying was assessed 30 min and 8 h later. A time-course study was also performed by injecting 50 µg of toxin-g /kg and measuring the effect on gastric emptying at times 0.25, 0.5, 1, 2, 4, 8, 24 and 48 h post-venom. Each envenomed animal was paired with its saline control and all received a saline test meal solution containing phenol red (60 µg/ml as a marker. Ten minutes after administering the test meal by gavage the animals were sacrificed and gastric retention was determined by measuring the residual marker concentration of the test meal. A significant delay in gastric emptying, at 30 min and 8 h post-venom, was observed only after 50 and 100 µg of toxin-g /kg compared to control values. The responses to these two doses were significantly different after 8 h post-venom. Toxin-g (50 µg/kg significantly delayed the gastric emptying of liquids at all times studied, with a peak response at 4 h after toxin administration compared to control values. These results indicate that the iv injection of toxin-g may induce a rapid, intense and sustained inhibition of gastric emptying 0.25 to 48 h after envenomation.

  9. Development of an antibiotic marker-free platform for heterologous protein production in Streptomyces.

    Science.gov (United States)

    Sevillano, Laura; Díaz, Margarita; Santamaría, Ramón I

    2017-09-26

    The industrial use of enzymes produced by microorganisms is continuously growing due to the need for sustainable solutions. Nevertheless, many of the plasmids used for recombinant production of proteins in bacteria are based on the use of antibiotic resistance genes as selection markers. The safety concerns and legal requirements surrounding the increased use of antibiotic resistance genes have made the development of new antibiotic-free approaches essential. In this work, a system completely free of antibiotic resistance genes and useful for the production of high yields of proteins in Streptomyces is described. This system is based on the separation of the two components of the yefM/yoeBsl (antitoxin/toxin) operon; the toxin (yoeBsl) gene, responsible for host death, is integrated into the genome and the antitoxin gene (yefMsl), which inactivates the toxin, is located in the expression plasmid. To develop this system, the toxin gene was integrated into the genome of a strain lacking the complete operon, and the antibiotic resistance gene integrated along with the toxin was eliminated by Cre recombinase to generate a final host strain free of any antibiotic resistance marker. In the same way, the antibiotic resistance gene from the final expression plasmid was removed by Dre recombinase. The usefulness of this system was analysed by checking the production of two hydrolases from different Streptomyces. Production of both proteins, with potential industrial use, was high and stable over time after strain storage and after serial subcultures. These results support the robustness and stability of the positive selection system developed. The total absence of antibiotic resistance genes makes this system a powerful tool for using Streptomyces as a host to produce proteins at the industrial level. This work is the first Streptomyces antibiotic marker-free system to be described. Graphical abstract Antibiotic marker-free platform for protein expression in Streptomyces

  10. Gene and antigen markers of Shiga-toxin producing E. coli from Michigan and Indiana river water: Occurrence and relation to recreational water quality criteria

    Science.gov (United States)

    Duris, J.W.; Haack, S.K.; Fogarty, L.R.

    2009-01-01

    The relation of bacterial pathogen occurrence to fecal indicator bacteria (FIB) concentrations used for recreational water quality criteria (RWQC) is poorly understood. This study determined the occurrence of Shiga-toxin producing Escherichia coli (STEC) markers and their relation to FIB concentrations in Michigan and Indiana river water. Using 67 fecal coliform (FC) bacteria cultures from 41 river sites in multiple watersheds, we evaluated the occurrence of five STEC markers: the Escherichia coli (EC) O157 antigen and gene, and the STEC virulence genes eaeA, stx1, and stx2. Simple isolations from selected FC cultures yielded viable EC O157. By both antigen and gene assays, EC O157 was detected in a greater proportion of samples exceeding rather than meeting FC RWQC (P public health decision-making. Copyright ?? 2009 by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America. All rights reserved.

  11. Cholera epidemics of the past offer new insights into an old enemy

    DEFF Research Database (Denmark)

    Phelps, Matthew David; Linnet Perner, Mads; Pitzer, Virginia E

    2018-01-01

    BACKGROUND: Although cholera is considered the quintessential long-cycle waterborne disease, studies have emphasized the existence of short-cycle (food, household) transmission. We investigated singular Danish cholera epidemics (1853) to elucidate epidemiological parameters and modes of spread...... intervals. RESULTS: Epidemics were seeded by travelers from cholera-affected cities; initial transmission chains involving household members and caretakers ensued. Cholera killed 3.4-8.9% of the populations, with highest mortality among seniors (16%) and lowest in children (2.7%). Transmissibility (R0...

  12. Evaluation of Knowledge and Practices Regarding Cholera, Water Treatment, Hygiene, and Sanitation before and after an Oral Cholera Vaccination Campaign—Haiti, 2013–2014

    Science.gov (United States)

    Childs, Lana; François, Jeannot; Choudhury, Alina; Wannemuehler, Kathleen; Dismer, Amber; Hyde, Terri B.; Yen, Catherine Y.; Date, Kashmira A.; Juin, Stanley; Katz, Mark A.; Kantor, Erica Felker; Routh, Janell; Etheart, Melissa; Wright, Tracie; Adrien, Paul; Tohme, Rania A.

    2016-01-01

    In 2013, the Government of Haiti implemented its first oral cholera vaccine (OCV) campaign in Petite Anse, an urban setting, and Cerca Carvajal, a rural commune. We conducted and compared responses to two independent cross-sectional knowledge and practices household surveys pre- (N = 297) and post- (N = 302) OCV campaign in Petite Anse. No significant differences in knowledge about causes, symptoms, and prevention of cholera were noted. Compared with precampaign respondents, fewer postcampaign respondents reported treating (66% versus 27%, P treatment practices necessary for cholera and other diarrheal diseases prevention were noted in the postcampaign survey. Future OCV campaigns in Haiti should be used as an opportunity to emphasize the importance of maintaining good water, sanitation, and hygiene practices, and include a comprehensive, integrated approach for cholera control. PMID:27799642

  13. Detection of viable toxigenic Vibrio cholerae and virulent Shigella ...

    African Journals Online (AJOL)

    . cholerae and the invasion plasmid antigen gene (ipaH) of virulent Shigella spp., was performed and the PCR products were visualised by agarose gel electrophoresis. The assay allowed the detection of as few as 1 cfu/100 ml of V. cholerae ...

  14. [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)

  15. Single-dose Live Oral Cholera Vaccine CVD 103-HgR Protects Against Human Experimental Infection With Vibrio cholerae O1 El Tor.

    Science.gov (United States)

    Chen, Wilbur H; Cohen, Mitchell B; Kirkpatrick, Beth D; Brady, Rebecca C; Galloway, David; Gurwith, Marc; Hall, Robert H; Kessler, Robert A; Lock, Michael; Haney, Douglas; Lyon, Caroline E; Pasetti, Marcela F; Simon, Jakub K; Szabo, Flora; Tennant, Sharon; Levine, Myron M

    2016-06-01

    No licensed cholera vaccine is presently available in the United States. Cholera vaccines available in other countries require 2 spaced doses. A single-dose cholera vaccine that can rapidly protect short-notice travelers to high-risk areas and help control explosive outbreaks where logistics render 2-dose immunization regimens impractical would be a major advance.PXVX0200, based on live attenuated Vibrio cholerae O1 classical Inaba vaccine strain CVD 103-HgR, elicits seroconversion of vibriocidal antibodies (a correlate of protection) within 10 days of a single oral dose. We investigated the protection conferred by this vaccine in a human cholera challenge model. Consenting healthy adult volunteers, 18-45 years old, were randomly allocated 1:1 to receive 1 oral dose of vaccine (approximately 5 × 10(8) colony-forming units [CFU]) or placebo in double-blind fashion. Volunteers ingested approximately 1 × 10(5) CFU of wild-type V. cholerae O1 El Tor Inaba strain N16961 10 days or 3 months after vaccination and were observed on an inpatient research ward for stool output measurement and management of hydration. The vaccine was well tolerated, with no difference in adverse event frequency among 95 vaccinees vs 102 placebo recipients. The primary endpoint, moderate (≥3.0 L) to severe (≥5.0 L) diarrheal purge, occurred in 39 of 66 (59.1%) placebo controls but only 2 of 35 (5.7%) vaccinees at 10 days (vaccine efficacy, 90.3%; P < .0001) and 4 of 33 (12.1%) vaccinees at 3 months (vaccine efficacy, 79.5%; P < .0001). The significant vaccine efficacy documented 10 days and 3 months after 1 oral dose of PXVX0200 supports further development as a single-dose cholera vaccine. NCT01895855. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  16. 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.

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

    Science.gov (United States)

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

    2014-01-01

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

  18. Genomic Epidemiology of Vibrio cholerae O1 Associated with Floods, Pakistan, 2010

    Science.gov (United States)

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

    2014-01-01

    In August 2010, Pakistan experienced major floods and a subsequent cholera epidemic. To clarify the population dynamics and transmission of Vibrio cholerae in Pakistan, we sequenced the genomes of all V. cholerae O1 El Tor isolates and compared the sequences to a global collection of 146 V. cholerae strains. Within the global phylogeny, all isolates from Pakistan formed 2 new subclades (PSC-1 and PSC-2), lying in the third transmission wave of the seventh-pandemic lineage that could be distinguished by signature deletions and their antimicrobial susceptibilities. Geographically, PSC-1 isolates originated from the coast, whereas PSC-2 isolates originated from inland areas flooded by the Indus River. Single-nucleotide polymorphism accumulation analysis correlated river flow direction with the spread of PSC-2. We found at least 2 sources of cholera in Pakistan during the 2010 epidemic and illustrate the value of a global genomic data bank in contextualizing cholera outbreaks. PMID:24378019

  19. A prolonged, community-wide cholera outbreak associated with drinking water contaminated by sewage in Kasese District, western Uganda.

    Science.gov (United States)

    Kwesiga, Benon; Pande, Gerald; Ario, Alex Riolexus; Tumwesigye, Nazarius Mbona; Matovu, Joseph K B; Zhu, Bao-Ping

    2017-07-18

    In May 2015, a cholera outbreak that had lasted 3 months and infected over 100 people was reported in Kasese District, Uganda, where multiple cholera outbreaks had occurred previously. We conducted an investigation to identify the mode of transmission to guide control measures. We defined a suspected case as onset of acute watery diarrhoea from 1 February 2015 onwards in a Kasese resident. A confirmed case was a suspected case with Vibrio cholerae O1 El Tor, serotype Inaba cultured from a stool sample. We reviewed medical records to find cases. We conducted a case-control study to compare exposures among confirmed case-persons and asymptomatic controls, matched by village and age-group. We conducted environmental assessments. We tested water samples from the most affected area for total coliforms using the Most Probable Number (MPN) method. We identified 183 suspected cases including 61 confirmed cases of Vibrio cholerae 01; serotype Inaba, with onset between February and July 2015. 2 case-persons died of cholera. The outbreak occurred in 80 villages and affected all age groups; the highest attack rate occurred in the 5-14 year age group (4.1/10,000). The outbreak started in Bwera Sub-County bordering the Democratic Republic of Congo and spread eastward through sustained community transmission. The first case-persons were involved in cross-border trading. The case-control study, which involved 49 confirmed cases and 201 controls, showed that 94% (46/49) of case-persons compared with 79% (160/201) of control-persons drank water without boiling or treatment (OR M-H =4.8, 95% CI: 1.3-18). Water collected from the two main sources, i.e., public pipes (consumed by 39% of case-persons and 38% of control-persons) or streams (consumed by 29% of case-persons and 24% control-persons) had high coliform counts, a marker of faecal contamination. Environmental assessment revealed evidence of open defecation along the streams. No food items were significantly associated with

  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. The case of cholera preparedness, response and prevention in the ...

    African Journals Online (AJOL)

    2011-10-07

    Oct 7, 2011 ... Keywords: Cholera prevention, preparedness and response, socio-political understanding of cholera, socio-cultural understanding .... cies of bacteria or viruses. ... quality such as boiling, chlorination, and filtration are not eco-.

  2. Cultural influences behind cholera transmission in the Far North ...

    African Journals Online (AJOL)

    Introduction: in recent years, the Far North Region of Cameroon has experienced serious and recurrent cholera outbreaks. Yet, understanding of cultural influences on outbreaks and spread remain poorly understood. This qualitative study explored cultural influences on cholera exposure in this region. Methods: interviews ...

  3. Antibiotic Resistance of Vibrio cholerae Isolates from Kashan, Iran

    Directory of Open Access Journals (Sweden)

    Afzali H.MD,

    2016-03-01

    Full Text Available Abstract Aims: Cholera is an acute diarrheal disease that can lead to severe dehydration and death. Antibiotic resistance is a big challenge in infective disease like Cholera. The present study aimed to understand the characteristics and trends of antibiotic resistance of V. cholerae isolations in and around Kashan, Iran. Instrument & Methods: In this descriptive cross-sectional study, samples were gathered using census method from 1998 to 2013 in Kashan, Iran. 1132 fecal samples of patients with acute diarrhea and 237 samples of suspected water samples were taken. The serotypes and biotypes were determined by an enzymatic method. Antibiotic susceptibility test was performed by using Disk Diffusion Method. Data were analyzed using SPSS 23 software. Fisher-exact and Chi-square tests were used to compare the statistical parameters. Findings: 96 fecal samples (8.5% and 18 water samples (7.6% were positive for Vibrio cholerae. Non-agglutinating (Nag isolates (75.4% were more common than serotype Inaba (13.2% and Ogawa (11.4%. Nag serotypes were mostly resistant to cefixime (44% and ampicillin (33%. In contaminated water samples also the most frequent cases were Nag serotype (50%. Nag serotype showed 22.2% of resistance to ampicillin and nitrofurantoin. Conclusion: Vibrio cholerae isolates in Kashan, Iran, are highly resistant to antibiotics, especially Nag serotypes.

  4. Investigating the role of water in the Diffusion of Cholera using Agent-Based simulation

    Science.gov (United States)

    Augustijn, Ellen-Wien; Doldersum, Tom; Augustijn, Denie

    2014-05-01

    Traditionally, cholera was considered to be a waterborne disease. Currently we know that many other factors can contribute to the spread of this disease including human mobility and human behavior. However, the hydrological component in cholera diffusion is significant. The interplay between cholera and water includes bacteria (V. cholera) that survive in the aquatic environment, the possibility that run-off water from dumpsites carries the bacteria to surface water (rivers and lakes), and when the bacteria reach streams they can be carried downstream to infect new locations. Modelling is a very important tool to build theory on the interplay between different types of transmission mechanisms that together are responsible for the spread of Cholera. Agent-based simulation models are very suitable to incorporate behavior at individual level and to reproduce emergence. However, it is more difficult to incorporate the hydrological components in this type of model. In this research we present the hydrological component of an Agent-Based Cholera model developed to study a Cholera epidemic in Kumasi (Ghana) in 2005. The model was calibrated on the relative contribution of each community to the distributed pattern of cholera rather than the absolute number of incidences. Analysis of the results shows that water plays an important role in the diffusion of cholera: 75% of the cholera cases were infected via river water that was contaminated by runoff from the dumpsites. To initiate infections upstream, the probability of environment-to-human transmission seemed to be overestimated compared to what may be expected from literature. Scenario analyses show that there is a strong relation between the epidemic curve and the rainfall. Removing dumpsites that are situated close to the river resulted in a strong decrease in the number of cholera cases. Results are sensitive to the scheduling of the daily activities and the survival time of the cholera bacteria.

  5. Environmental factor analysis of cholera in China using remote sensing and geographical information systems.

    Science.gov (United States)

    Xu, M; Cao, C X; Wang, D C; Kan, B; Xu, Y F; Ni, X L; Zhu, Z C

    2016-04-01

    Cholera is one of a number of infectious diseases that appears to be influenced by climate, geography and other natural environments. This study analysed the environmental factors of the spatial distribution of cholera in China. It shows that temperature, precipitation, elevation, and distance to the coastline have significant impact on the distribution of cholera. It also reveals the oceanic environmental factors associated with cholera in Zhejiang, which is a coastal province of China, using both remote sensing (RS) and geographical information systems (GIS). The analysis has validated the correlation between indirect satellite measurements of sea surface temperature (SST), sea surface height (SSH) and ocean chlorophyll concentration (OCC) and the local number of cholera cases based on 8-year monthly data from 2001 to 2008. The results show the number of cholera cases has been strongly affected by the variables of SST, SSH and OCC. Utilizing this information, a cholera prediction model has been established based on the oceanic and climatic environmental factors. The model indicates that RS and GIS have great potential for designing an early warning system for cholera.

  6. Fuzzy expert systems and GIS for cholera health risk prediction in southern Africa

    CSIR Research Space (South Africa)

    Fleming, GJ

    2007-01-01

    Full Text Available Cholera (Vibrio cholerae) is endemic in southern Africa and frequently breaks out in epidemics along the eastern seaboard. Extensive resources are directed at combating cholera yet it remains a significant problem. Limited resources could better...

  7. Historical epidemiology of the second cholera pandemic: relevance to present day disease dynamics.

    Directory of Open Access Journals (Sweden)

    Christina H Chan

    Full Text Available Despite nearly two centuries of study, the fundamental transmission dynamic properties of cholera remain incompletely characterized. We used historical time-series data on the spread of cholera in twelve European and North American cities during the second cholera pandemic, as reported in Amariah Brigham's 1832 A Treatise on Epidemic Cholera, to parameterize simple mathematical models of cholera transmission. Richards growth models were used to derive estimates of the basic reproductive number (R0 (median: 16.0, range: 1.9 to 550.9 and the proportion of unrecognized cases (mean: 96.3%, SD: 0.04%. Heterogeneity in model-generated R0 estimates was consistent with variability in cholera dynamics described by contemporary investigators and may represent differences in the nature of cholera spread. While subject to limitations associated with measurement and the absence of microbiological diagnosis, historical epidemic data are a potentially rich source for understanding pathogen dynamics in the absence of control measures, particularly when used in conjunction with simple and readily interpretable mathematical models.

  8. Spatially explicit modelling of cholera epidemics

    Science.gov (United States)

    Finger, F.; Bertuzzo, E.; Mari, L.; Knox, A. C.; Gatto, M.; Rinaldo, A.

    2013-12-01

    Epidemiological models can provide crucial understanding about the dynamics of infectious diseases. Possible applications range from real-time forecasting and allocation of health care resources to testing alternative intervention mechanisms such as vaccines, antibiotics or the improvement of sanitary conditions. We apply a spatially explicit model to the cholera epidemic that struck Haiti in October 2010 and is still ongoing. The dynamics of susceptibles as well as symptomatic and asymptomatic infectives are modelled at the scale of local human communities. Dissemination of Vibrio cholerae through hydrological transport and human mobility along the road network is explicitly taken into account, as well as the effect of rainfall as a driver of increasing disease incidence. The model is calibrated using a dataset of reported cholera cases. We further model the long term impact of several types of interventions on the disease dynamics by varying parameters appropriately. Key epidemiological mechanisms and parameters which affect the efficiency of treatments such as antibiotics are identified. Our results lead to conclusions about the influence of different intervention strategies on the overall epidemiological dynamics.

  9. A global map of suitability for coastal Vibrio cholerae under current and future climate conditions.

    Science.gov (United States)

    Escobar, Luis E; Ryan, Sadie J; Stewart-Ibarra, Anna M; Finkelstein, Julia L; King, Christine A; Qiao, Huijie; Polhemus, Mark E

    2015-09-01

    Vibrio cholerae is a globally distributed water-borne pathogen that causes severe diarrheal disease and mortality, with current outbreaks as part of the seventh pandemic. Further understanding of the role of environmental factors in potential pathogen distribution and corresponding V. cholerae disease transmission over time and space is urgently needed to target surveillance of cholera and other climate and water-sensitive diseases. We used an ecological niche model (ENM) to identify environmental variables associated with V. cholerae presence in marine environments, to project a global model of V. cholerae distribution in ocean waters under current and future climate scenarios. We generated an ENM using published reports of V. cholerae in seawater and freely available remotely sensed imagery. Models indicated that factors associated with V. cholerae presence included chlorophyll-a, pH, and sea surface temperature (SST), with chlorophyll-a demonstrating the greatest explanatory power from variables selected for model calibration. We identified specific geographic areas for potential V. cholerae distribution. Coastal Bangladesh, where cholera is endemic, was found to be environmentally similar to coastal areas in Latin America. In a conservative climate change scenario, we observed a predicted increase in areas with environmental conditions suitable for V. cholerae. Findings highlight the potential for vulnerability maps to inform cholera surveillance, early warning systems, and disease prevention and control. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  10. Response to the cholera outbreak in South Sudan

    African Journals Online (AJOL)

    On Thursday, May 15th 2014, the Ministry of Health (MoH) of the Republic of South Sudan declared a cholera outbreak in the capital Juba. As we go to press, the cholera has spread to other parts of the country and the cases are increasing. In its press statement, the MoH said it had “Reactivated a national emergency ...

  11. An Ecosocial Approach to the Epidemic of Cholera in the Marshall Islands

    Directory of Open Access Journals (Sweden)

    Wesley Palmer

    2007-04-01

    Full Text Available Abstract: A cholera outbreak occurred in the Marshall Islands in December 2000 to January 2001 with over 400 cases and six deaths. Within Kwajalein Atoll, cholera occurred on Ebeye Island, while it did not occur on Kwajalein Island, three miles away. We apply Krieger’s ecosocial approach in order to explicate the reasons for this dichotomy. We first examine how Marshallese people came to embody cholera as a disease state. Secondly, we examine the (a arrangements of power, property, production, and consumption in the Ebeye-Kwajalein complex, as well as (b human biology as it has been shaped by the ecological context in order to elucidate the pathways to the embodiment of cholera. Thirdly, we examine the cumulative interplay between exposure to cholera, as well as susceptibility and resistance to the disease at the level of individuals and the island-wide level. Fourthly, we examine who is responsible for the cholera outbreak and who describes the phenomena. We conclude that the outbreak of cholera in the Marshall Islands can be considered the biologic embodiment of disparate political and economic conditions and ecological imbalance. We suggest courses of action for those interested in addressing the inequalities and working towards health.

  12. Linking Satellite Derived Land Surface Temperature with Cholera: A Case Study for South Sudan

    Science.gov (United States)

    Aldaach, H. S. V.; Jutla, A.; Akanda, A. S.; Colwell, R. R.

    2014-12-01

    A sudden onset of cholera in South Sudan, in April 2014 in Northern Bari in Juba town resulted in more than 400 cholera cases after four weeks of initial outbreak with a case of fatality rate of CFR 5.4%. The total number of reported cholera cases for the period of April to July, 2014 were 5,141 including 114 deaths. With the limited efficacy of cholera vaccines, it is necessary to develop mechanisms to predict cholera occurrence and thereafter devise intervention strategies for mitigating impacts of the disease. Hydroclimatic processes, primarily precipitation and air temperature are related to epidemic and episodic outbreak of cholera. However, due to coarse resolution of both datasets, it is not possible to precisely locate the geographical location of disease. Here, using Land Surface Temperature (LST) from MODIS sensors, we have developed an algorithm to identify regions susceptible for cholera. Conditions for occurrence of cholera were detectable at least one month in advance in South Sudan and were statistically sensitive to hydroclimatic anomalies of land surface and air temperature, and precipitation. Our results indicate significant spatial and temporal averaging required to infer usable information from LST over South Sudan. Preliminary results that geographically location of cholera outbreak was identifiable within 1km resolution of the LST data.

  13. Risk Map of Cholera Infection for Vaccine Deployment: The Eastern Kolkata Case

    Science.gov (United States)

    You, Young Ae; Ali, Mohammad; Kanungo, Suman; Sah, Binod; Manna, Byomkesh; Puri, Mahesh; Nair, G. Balakrish; Bhattacharya, Sujit Kumar; Convertino, Matteo; Deen, Jacqueline L.; Lopez, Anna Lena; Wierzba, Thomas F.; Clemens, John; Sur, Dipika

    2013-01-01

    Background Despite advancement of our knowledge, cholera remains a public health concern. During March-April 2010, a large cholera outbreak afflicted the eastern part of Kolkata, India. The quantification of importance of socio-environmental factors in the risk of cholera, and the calculation of the risk is fundamental for deploying vaccination strategies. Here we investigate socio-environmental characteristics between high and low risk areas as well as the potential impact of vaccination on the spatial occurrence of the disease. Methods and Findings The study area comprised three wards of Kolkata Municipal Corporation. A mass cholera vaccination campaign was conducted in mid-2006 as the part of a clinical trial. Cholera cases and data of the trial to identify high risk areas for cholera were analyzed. We used a generalized additive model (GAM) to detect risk areas, and to evaluate the importance of socio-environmental characteristics between high and low risk areas. During the one-year pre-vaccination and two-year post-vaccination periods, 95 and 183 cholera cases were detected in 111,882 and 121,827 study participants, respectively. The GAM model predicts that high risk areas in the west part of the study area where the outbreak largely occurred. High risk areas in both periods were characterized by poor people, use of unsafe water, and proximity to canals used as the main drainage for rain and waste water. Cholera vaccine uptake was significantly lower in the high risk areas compared to low risk areas. Conclusion The study shows that even a parsimonious model like GAM predicts high risk areas where cholera outbreaks largely occurred. This is useful for indicating where interventions would be effective in controlling the disease risk. Data showed that vaccination decreased the risk of infection. Overall, the GAM-based risk map is useful for policymakers, especially those from countries where cholera remains to be endemic with periodic outbreaks. PMID:23936491

  14. 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

  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....... Alternatively, it has been proposed that bacterial pathogens are an integral part of the natural microbial food web and thus their survival is constrained by protozoan predation. Here, we report that both explanations are interrelated. our data show that biofilms are the protective agent enabling V. cholerae...... to survive protozoan grazing while their planktonic counterparts are eliminated. Grazing on planktonic V. cholerae was found to select for the biofilm-enhancing rugose phase variant, which is adapted to the surf ace-associated niche by the production of exopolymers. Interestingly, grazing resistance in V...

  16. Cost-benefit comparisons of investments in improved water supply and cholera vaccination programs.

    Science.gov (United States)

    Jeuland, Marc; Whittington, Dale

    2009-05-18

    This paper presents the first cost-benefit comparison of improved water supply investments and cholera vaccination programs. Specifically, we compare two water supply interventions -- deep wells with public hand pumps and biosand filters (an in-house, point-of-use water treatment technology) -- with two types of cholera immunization programs with new-generation vaccines -- general community-based and targeted and school-based programs. In addition to these four stand-alone investments, we also analyze five combinations of water and vaccine interventions: (1) borehole+hand pump and community-based cholera vaccination, (2) borehole+hand pump and school-based cholera vaccination, (3) biosand filter and community-based cholera vaccination, (4) biosand filter and school-based cholera vaccination, and (5) biosand filter and borehole+hand pump. Using recent data applicable to developing country locations for parameters such as disease incidence, the effectiveness of vaccine and water supply interventions against diarrheal diseases, and the value of a statistical life, we construct cost-benefit models for evaluating these interventions. We then employ probabilistic sensitivity analysis to estimate a frequency distribution of benefit-cost ratios for all four interventions, given a wide variety of possible parameter combinations. Our results demonstrate that there are many plausible conditions in developing countries under which these interventions will be attractive, but that the two improved water supply interventions and the targeted cholera vaccination program are much more likely to yield attractive cost-benefit outcomes than a community-based vaccination program. We show that implementing community-based cholera vaccination programs after borehole+hand pump or biosand filters have already been installed will rarely be justified. This is especially true when the biosand filters are already in place, because these achieve substantial cholera risk reductions on their own

  17. Comparative effectiveness of different strategies of oral cholera vaccination in bangladesh: a modeling study.

    Directory of Open Access Journals (Sweden)

    Dobromir T Dimitrov

    2014-12-01

    Full Text Available Killed, oral cholera vaccines have proven safe and effective, and several large-scale mass cholera vaccination efforts have demonstrated the feasibility of widespread deployment. This study uses a mathematical model of cholera transmission in Bangladesh to examine the effectiveness of potential vaccination strategies.We developed an age-structured mathematical model of cholera transmission and calibrated it to reproduce the dynamics of cholera in Matlab, Bangladesh. We used the model to predict the effectiveness of different cholera vaccination strategies over a period of 20 years. We explored vaccination programs that targeted one of three increasingly focused age groups (the entire vaccine-eligible population of age one year and older, children of ages 1 to 14 years, or preschoolers of ages 1 to 4 years and that could occur either as campaigns recurring every five years or as continuous ongoing vaccination efforts. Our modeling results suggest that vaccinating 70% of the population would avert 90% of cholera cases in the first year but that campaign and continuous vaccination strategies differ in effectiveness over 20 years. Maintaining 70% coverage of the population would be sufficient to prevent sustained transmission of endemic cholera in Matlab, while vaccinating periodically every five years is less effective. Selectively vaccinating children 1-14 years old would prevent the most cholera cases per vaccine administered in both campaign and continuous strategies.We conclude that continuous mass vaccination would be more effective against endemic cholera than periodic campaigns. Vaccinating children averts more cases per dose than vaccinating all age groups, although vaccinating only children is unlikely to control endemic cholera in Bangladesh. Careful consideration must be made before generalizing these results to other regions.

  18. Comparative effectiveness of different strategies of oral cholera vaccination in bangladesh: a modeling study.

    Science.gov (United States)

    Dimitrov, Dobromir T; Troeger, Christopher; Halloran, M Elizabeth; Longini, Ira M; Chao, Dennis L

    2014-12-01

    Killed, oral cholera vaccines have proven safe and effective, and several large-scale mass cholera vaccination efforts have demonstrated the feasibility of widespread deployment. This study uses a mathematical model of cholera transmission in Bangladesh to examine the effectiveness of potential vaccination strategies. We developed an age-structured mathematical model of cholera transmission and calibrated it to reproduce the dynamics of cholera in Matlab, Bangladesh. We used the model to predict the effectiveness of different cholera vaccination strategies over a period of 20 years. We explored vaccination programs that targeted one of three increasingly focused age groups (the entire vaccine-eligible population of age one year and older, children of ages 1 to 14 years, or preschoolers of ages 1 to 4 years) and that could occur either as campaigns recurring every five years or as continuous ongoing vaccination efforts. Our modeling results suggest that vaccinating 70% of the population would avert 90% of cholera cases in the first year but that campaign and continuous vaccination strategies differ in effectiveness over 20 years. Maintaining 70% coverage of the population would be sufficient to prevent sustained transmission of endemic cholera in Matlab, while vaccinating periodically every five years is less effective. Selectively vaccinating children 1-14 years old would prevent the most cholera cases per vaccine administered in both campaign and continuous strategies. We conclude that continuous mass vaccination would be more effective against endemic cholera than periodic campaigns. Vaccinating children averts more cases per dose than vaccinating all age groups, although vaccinating only children is unlikely to control endemic cholera in Bangladesh. Careful consideration must be made before generalizing these results to other regions.

  19. A model to predict when a cholera outbreak might hit the Congo

    Science.gov (United States)

    Schultz, Colin

    2014-09-01

    In 2011, as many as 600,000 people in 58 countries contracted cholera, with thousands succumbing to the disease. In most countries, cholera is rare. In others, like the Democratic Republic of the Congo, cholera is an endemic threat, always lurking in the background waiting for the right set of conditions to spark an outbreak.

  20. [Natural toxins in inter- and intraspecies interaction of human being (elements of ethnotoxinology)].

    Science.gov (United States)

    Gelashvili, D B

    2002-01-01

    The author considers the application of natural toxins as arrow poison by Homo sapiens from ancient time till today for hunting and ethnic wars on the example of natives of Asia, Africa, South America and Oceania. Geographic isolation was important determining the spectrum of natural toxin sources and the methods of their application. Cellular and molecular mechanisms of arrow poisons effects are considered in biogeographical context: aconitin and strychnin in Asia, diamphotoxin in Africa, indole alcaloids of plants and steroid alcaloids of amphibian in Central and South America, palytoxin in Oceania islands. High efficiency and selective effect of natural toxins allow to use them as molecular markers in current studies of functional membrane architecture and cellular structures. Great differences in pace of civilization development leads to the co-existence at the beginning of the XXI century ethnic groups that use natural toxins as arrow poison and human beings that use the same toxins in fundamental and applied investigations within international scientific society.

  1. Characterization of ultraviolet light-induced diphtheria toxin-resistant mutations in normal and Xeroderma pigmentosum human fibroblasts

    International Nuclear Information System (INIS)

    Glover, T.W.

    1979-01-01

    Quantitative mutagenesis studies in human cells have been severely limited by the lack of reliable genetic markers. Experiments were therefore performed to develop and characterize a better quantitative mutation assay for human cells. The uv-induction of diphtheria toxin resistant (DT/sup r/) mutations in normal and excision repair defective xeroderma pigmentosum (XP) fibroblasts has been quantitatively characterized. A concentration of diphtheria toxin to use in the selection of resistant mutants was determined whereby DT/sup r/ cells are cross-resistant to Pseudomonas aeurginosa exotoxin A, indicating mutants have altered elongation factor-2 (EF-2) which is not susceptible to ADP-ribosylation by either toxin. Results of this study indicate that XP fibroblasts have higher uv-induced mutation frequencies per unit uv-dose but similar frequencies per unit survival compared to normal cells as measured using a new genetic marker for quantitative mutagenesis. Furthermore, these results support a prediction of the mutation theory of cancer, namely, that cells from individuals with certain human syndromes that predispose the individual to cancer will have higher induced mutation frequencies than cells from non-susceptible individuals. This newly characterized genetic marker should be useful in quantitative mutagenesis studies in human cells

  2. Model of Cholera Forecasting Using Artificial Neural Network in Chabahar City, Iran

    Directory of Open Access Journals (Sweden)

    Zahra Pezeshki

    2016-02-01

    Full Text Available Background: Cholera as an endemic disease remains a health issue in Iran despite decrease in incidence. Since forecasting epidemic diseases provides appropriate preventive actions in disease spread, different forecasting methods including artificial neural networks have been developed to study parameters involved in incidence and spread of epidemic diseases such as cholera. Objectives: In this study, cholera in rural area of Chabahar, Iran was investigated to achieve a proper forecasting model. Materials and Methods: Data of cholera was gathered from 465 villages, of which 104 reported cholera during ten years period of study. Logistic regression modeling and correlate bivariate were used to determine risk factors and achieve possible predictive model one-hidden-layer perception neural network with backpropagation training algorithm and the sigmoid activation function was trained and tested between the two groups of infected and non-infected villages after preprocessing. For determining validity of prediction, the ROC diagram was used. The study variables included climate conditions and geographical parameters. Results: After determining significant variables of cholera incidence, the described artificial neural network model was capable of forecasting cholera event among villages of test group with accuracy up to 80%. The highest accuracy was achieved when model was trained with variables that were significant in statistical analysis describing that the two methods confirm the result of each other. Conclusions: Application of artificial neural networking assists forecasting cholera for adopting protective measures. For a more accurate prediction, comprehensive information is required including data on hygienic, social and demographic parameters.

  3. Implementation research: reactive mass vaccination with single-dose oral cholera vaccine, Zambia.

    Science.gov (United States)

    Poncin, Marc; Zulu, Gideon; Voute, Caroline; Ferreras, Eva; Muleya, Clara Mbwili; Malama, Kennedy; Pezzoli, Lorenzo; Mufunda, Jacob; Robert, Hugues; Uzzeni, Florent; Luquero, Francisco J; Chizema, Elizabeth; Ciglenecki, Iza

    2018-02-01

    To describe the implementation and feasibility of an innovative mass vaccination strategy - based on single-dose oral cholera vaccine - to curb a cholera epidemic in a large urban setting. In April 2016, in the early stages of a cholera outbreak in Lusaka, Zambia, the health ministry collaborated with Médecins Sans Frontières and the World Health Organization in organizing a mass vaccination campaign, based on single-dose oral cholera vaccine. Over a period of 17 days, partners mobilized 1700 health ministry staff and community volunteers for community sensitization, social mobilization and vaccination activities in 10 townships. On each day, doses of vaccine were delivered to vaccination sites and administrative coverage was estimated. Overall, vaccination teams administered 424 100 doses of vaccine to an estimated target population of 578 043, resulting in an estimated administrative coverage of 73.4%. After the campaign, few cholera cases were reported and there was no evidence of the disease spreading within the vaccinated areas. The total cost of the campaign - 2.31 United States dollars (US$) per dose - included the relatively low cost of local delivery - US$ 0.41 per dose. We found that an early and large-scale targeted reactive campaign using a single-dose oral vaccine, organized in response to a cholera epidemic within a large city, to be feasible and appeared effective. While cholera vaccines remain in short supply, the maximization of the number of vaccines in response to a cholera epidemic, by the use of just one dose per member of an at-risk community, should be considered.

  4. [The clinico-epidemiological characteristics of cholera patients in Mexico City].

    Science.gov (United States)

    Vilchis-Guizar, A E; Uribe-Márquez, S; Pérez-Sánchez, P L

    1999-01-01

    To compare the clinical and epidemiological characteristics of cholera patients and patients with diarrhea of different etiology (controls), treated at Mexican Institute of Social Security medical units in northeast Mexico City, from 1995 to 1998. Cross-sectional analytical study design. Data were collected using the official form "Immediate Notification of Cholera Cases" that each medical unit reports to the Coordination of Community Health. Statistical analysis consisted of comparisons of means and proportions between both groups. In 4,952 notifications, there were 588 cholera cases and 4,364 controls. The age range of cases was 39 to 51 years and 54% were females. During 1995 and 1997 (years with higher proportion of cases) patients with cholera had a greater frequency of watery evacuations than controls (97% vs. 73%), "rice water" appearance (31% vs. 13%), vomiting (72% vs. 63%), muscular spasms (49% vs. 26%), dehydration (83% vs. 71%), hypovolemic shock (10% vs. 1%) and death (0.85% vs. 0.25%). These differences were statistically significant. Cholera presents a biannual cyclic behavior; greater frequencies are associated with greater severity and complications. It is necessary to increase epidemiologic surveillance and medical efforts for opportune diagnosis and treatment.

  5. The role of socioeconomic status in longitudinal trends of cholera in Matlab, Bangladesh, 1993-2007.

    Science.gov (United States)

    Root, Elisabeth Dowling; Rodd, Joshua; Yunus, Mohammad; Emch, Michael

    2013-01-01

    There has been little evidence of a decline in the global burden of cholera in recent years as the number of cholera cases reported to WHO continues to rise. Cholera remains a global threat to public health and a key indicator of lack of socioeconomic development. Overall socioeconomic development is the ultimate solution for control of cholera as evidenced in developed countries. However, most research has focused on cross-county comparisons so that the role of individual- or small area-level socioeconomic status (SES) in cholera dynamics has not been carefully studied. Reported cases of cholera in Matlab, Bangladesh have fluctuated greatly over time and epidemic outbreaks of cholera continue, most recently with the introduction of a new serotype into the region. The wealth of longitudinal data on the population of Matlab provides a unique opportunity to explore the impact of socioeconomic status and other demographic characteristics on the long-term temporal dynamics of cholera in the region. In this population-based study we examine which factors impact the initial number of cholera cases in a bari at the beginning of the 0139 epidemic and the factors impacting the number of cases over time. Cholera data were derived from the ICDDR,B health records and linked to socioeconomic and geographic data collected as part of the Matlab Health and Demographic Surveillance System. Longitudinal zero-inflated Poisson (ZIP) multilevel regression models are used to examine the impact of environmental and socio-demographic factors on cholera counts across baris. Results indicate that baris with a high socioeconomic status had lower initial rates of cholera at the beginning of the 0139 epidemic (γ(01) = -0.147, p = 0.041) and a higher probability of reporting no cholera cases (α(01) = 0.156, p = 0.061). Populations in baris characterized by low SES are more likely to experience higher cholera morbidity at the beginning of an epidemic than populations in high

  6. Expression of cholera toxin B-proinsulin fusion protein in lettuce and tobacco chloroplasts--oral administration protects against development of insulitis in non-obese diabetic mice.

    Science.gov (United States)

    Ruhlman, Tracey; Ahangari, Raheleh; Devine, Andrew; Samsam, Mohtahsem; Daniell, Henry

    2007-07-01

    Lettuce and tobacco chloroplast transgenic lines expressing the cholera toxin B subunit-human proinsulin (CTB-Pins) fusion protein were generated. CTB-Pins accumulated up to ~16% of total soluble protein (TSP) in tobacco and up to ~2.5% of TSP in lettuce. Eight milligrams of powdered tobacco leaf material expressing CTB-Pins or, as negative controls, CTB-green fluorescent protein (CTB-GFP) or interferon-GFP (IFN-GFP), or untransformed leaf, were administered orally, each week for 7 weeks, to 5-week-old female non-obese diabetic (NOD) mice. The pancreas of CTB-Pins-treated mice showed decreased infiltration of cells characteristic of lymphocytes (insulitis); insulin-producing beta-cells in the pancreatic islets of CTB-Pins-treated mice were significantly preserved, with lower blood or urine glucose levels, by contrast with the few beta-cells remaining in the pancreatic islets of the negative controls. Increased expression of immunosuppressive cytokines, such as interleukin-4 and interleukin-10 (IL-4 and IL-10), was observed in the pancreas of CTB-Pins-treated NOD mice. Serum levels of immunoglobulin G1 (IgG1), but not IgG2a, were elevated in CTB-Pins-treated mice. Taken together, T-helper 2 (Th2) lymphocyte-mediated oral tolerance is a likely mechanism for the prevention of pancreatic insulitis and the preservation of insulin-producing beta-cells. This is the first report of expression of a therapeutic protein in transgenic chloroplasts of an edible crop. Transplastomic lettuce plants expressing CTB-Pins grew normally and transgenes were maternally inherited in T(1) progeny. This opens up the possibility for the low-cost production and delivery of human therapeutic proteins, and a strategy for the treatment of various other autoimmune diseases.

  7. 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

  8. [Cholera in children. A report of 8 cases].

    Science.gov (United States)

    Lezama-Basulto, L A; Mota-Hernández, F; Bravo-Barrios, E

    1993-11-01

    Cholera is an acute intestinal infection caused by Vibrio cholerae 01. When an infected person presents severe dehydration and is not adequately treated, he or she will develop hypovolemic shock and eventually could died. There is scarce information concerning this disease in the Pediatric group. Herein we report on eight cases of Pediatric cholera, in children 17 month to four years of age. Seven patients out of eight were admitted presenting dehydration. Four presenting mild or moderate dehydration and three presenting hypovolemic shock. These three patients were rehydrated by intravenous route and thereafter the hydration was maintained by oral therapy. The outcome was uneventful in six patients. One patient developed abdominal distention probably due to hypopotassemia, and another patient presented hyponatremia and seizures. All the patients recovered within five days after admission.

  9. Cholera outbreak in districts around Lake Chilwa, Malawi: Lessons ...

    African Journals Online (AJOL)

    Cholera is endemic in Malawi with seasonal outbreaks during the wet season. People living around Lake Chilwa rely on the lake for their water supply. From May 2009 to May 2010, a cholera outbreak occurred in fishing communities around Lake Chilwa. This paper describes the outbreak response and lessons learned for ...

  10. The cholera epidemic in South Africa, 1980 - 1987 Epidemiological ...

    African Journals Online (AJOL)

    During the cholera epidemic in South Africa, 1980-1987, 25251 cases of cholera were bacteriologically proven. The case-fatality rate was 1,4%. Outbreaks occurred in the summer rainfall season. Age-specific aUack rates followed the pattern typically found during the 'epidemic phase' of the disease in most years. The vast ...

  11. Association between earthquake events and cholera outbreaks: a cross-country 15-year longitudinal analysis.

    Science.gov (United States)

    Sumner, Steven A; Turner, Elizabeth L; Thielman, Nathan M

    2013-12-01

    Large earthquakes can cause population displacement, critical sanitation infrastructure damage, and increased threats to water resources, potentially predisposing populations to waterborne disease epidemics such as cholera. Problem The risk of cholera outbreaks after earthquake disasters remains uncertain. A cross-country analysis of World Health Organization (WHO) cholera data that would contribute to this discussion has yet to be published. A cross-country longitudinal analysis was conducted among 63 low- and middle-income countries from 1995-2009. The association between earthquake disasters of various effect sizes and a relative spike in cholera rates for a given country was assessed utilizing fixed-effects logistic regression and adjusting for gross domestic product per capita, water and sanitation level, flooding events, percent urbanization, and under-five child mortality. Also, the association between large earthquakes and cholera rate increases of various degrees was assessed. Forty-eight of the 63 countries had at least one year with reported cholera infections during the 15-year study period. Thirty-six of these 48 countries had at least one earthquake disaster. In adjusted analyses, country-years with ≥10,000 persons affected by an earthquake had 2.26 times increased odds (95 CI, 0.89-5.72, P = .08) of having a greater than average cholera rate that year compared to country-years having earthquake. The association between large earthquake disasters and cholera infections appeared to weaken as higher levels of cholera rate increases were tested. A trend of increased risk of greater than average cholera rates when more people were affected by an earthquake in a country-year was noted. However these findings did not reach statistical significance at traditional levels and may be due to chance. Frequent large-scale cholera outbreaks after earthquake disasters appeared to be relatively uncommon.

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

    Science.gov (United States)

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

    2017-02-01

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

  13. [Performance of Cholera-SMART and Pathogen-Detection-Kit in the quick diagnosis of cholera].

    Science.gov (United States)

    Bolaños, Hilda María; Acuña, María Teresa; Serrano, Ana María; Obando, Xinia; Mairena, Hazel; Cháves, Lorena; Sandí, Flor; Rodríguez, Gina; Tamplin, Mark L; Pérez, Enrique; Campos, Elena

    2004-10-01

    To compare the performance of two rapid systems for the diagnosis of cholera with the culture method, and to propose a strategy for improving the specificity and sensitivity of these systems and reducing the costs involved in making a diagnosis. The following institutions participated in the study: the National Bacteriology Referral Center (Centro Nacional de Referencia en Bacteriologia, CNRB) of the Costa Rican Institute for Research and Teaching in Nutrition and Health (Instituto Costarricense de Investigacion y Ensenanza en Nutricion y Salud, INCIENSA) and various hospitals in the provinces of Alajuela, Guanacaste and San Jose, in Costa Rica. A total of 237 feces samples were used to asses the performance of two tests for the rapid detection of Vibrio cholerae 01: the Pathogen Detection Kit (PDK, Intelligent Monitoring Systems, Gainesville, Florida, USA) and Cholera-SMART (New Horizons Diagnostics Corp., Columbia, Maryland, USA), both when applied directly (direct SMART and direct PDK) and when applied to specimens cultured in broth-enriched medium for 6 hours (SMART-6 and CPK-6) and for 18 hours (SMART-18 and PDK-18) at 37 degrees C in alkaline peptone water. Liquid and partially formed stools were cultured and examined by means of the rapid direct test; when the initial result was negative, the tests were repeated after culture for periods of 6 and 18 hours. Rectal and fecal swabs were obtained from feces cultured in enriched-broth medium for 6 and 18 hours. In addition, we studied the sensitivity of the rapid testing systems by using pure cultures of V. cholerae 01 (strain SOS-833, CNRB, Costa Rica) that were incubated for 18 to 24 hours, and we assessed the usefulness of observing motility under the microscope in order to rationalize the use of rapid methods. The sensitivity of the direct SMART test and of the direct PDK test was 100% when samples obtained from liquid and partially formed stools and from the intestinal contents of dead bodies were used. With

  14. The "cholera cloud" in the nineteenth-century "British World": history of an object-without-an-essence.

    Science.gov (United States)

    Mukharji, Projit Bihari

    2012-01-01

    The "cholera cloud" is one of the most persistent presences in the archives of nineteenth-century cholera in the "British World." Yet it has seldom received anything more than a passing acknowledgment from historians of cholera. Tracing the history of the cholera cloud as an object promises to open up a new dimension of the historically contingent experience of cholera, as well as make a significant contribution to the emergent literature on "thing theory." By conceptualizing the cholera cloud as an object-without-an-essence, this article demonstrates how global cholera pandemics in the nineteenth century produced globalized objects in which a near-universal recognizability and an utterly context-specific set of meanings, visions, and realities could ironically cohabit.

  15. Isothermal Titration Calorimeter For Research And Education (DURIP-10)

    Science.gov (United States)

    2011-05-12

    Plamen Atanassov, University of New Mexico , Shelley Minteer, Saint Louis University, and Scott Calabrese-Barton, Michigan State University. The...associated proteins will also be explored as targets including the ricin toxin B, cholera toxin B, and botulinum neurotoxin type E. The binding...domain from V. cholerae called VCA0042/PlzD. These measurements showed that cdG binding to VCA0042 is entropically unfavorable. They also

  16. An outbreak of cholera in Medipally village, Andhra Pradesh, India, 2013

    OpenAIRE

    Uthappa, Chengappa K.; Allam, Ramesh R.; Nalini, Chava; Gunti, Deepak; Udaragudi, Prasada R.; Tadi, Geetha P.; Murhekar, Manoj V.

    2015-01-01

    Background Cholera continues to remain endemic in over 50 countries and has caused large epidemics with around 3?5 million cases occurring every year in Asia alone. In India, cholera is endemic in many states. However, etiological information and age-specific incidence related to cholera outbreaks is limited. In November 2013, district authorities reported a cluster of diarrheal disease among residents of Medipally to the state surveillance unit. We investigated this cluster to confirm its et...

  17. [The in vitro action of plants on Vibrio cholerae].

    Science.gov (United States)

    Guevara, J M; Chumpitaz, J; Valencia, E

    1994-01-01

    Natural products of several plants, according to the geographic location, are used by Peruvian people in the popular treatment of diarrhea, with good success. When cholerae cases appeared in Peru, we were interested to know the "in vitro" effect against Vibrio cholerae 01, of these useful plants to treat diarrhea. The following plants were tested: Cichorium intybus, Althaea officinalis, Psorela glandulosa, Geranium maculatum, Punica granatum, Malus sativa, Cydonia oblonga, Chenopodium ambrosoides, Krameria triandria, Tea chinensis, Daucus carota, Persea gratissima, Psidium guayaba and Lippia dulcis. Decoction or infusion of the plants were used in the "in vitro" experiments. The following plants showed no "in vitro" effect against V. cholerae: Cichorium intybus, Althaea officinalis, Psorela glandulosa, Geranium maculatum, Chenopodium ambrosoides, Krameria triandria, Psidium guayaba, Lippia dulcis and Daucus carota. Decoction of Malus sativa and Cydenia oblonga showed bactericidal effect for their acidity and stone avocado (Persea gratissima) a late bactericidal effect. Tea infusión and the decoction of Punica granatum peel, showed the best bactericidal effect and we suggest to use them as to stop cholera spreading.

  18. Hydroclimatic mechanisms of cholera transmission in the Bengal Delta

    Science.gov (United States)

    Tretkoff, Ernie

    2011-07-01

    Cholera, a deadly waterborne disease, remains a major threat in many areas of the world, including the Bengal Delta region. In this region, cholera outbreaks have two annual peaks; the first occurs during the dry season in the spring, and the second occurs in the fall following the wet season. However, the large-scale hydroclimatic processes underlying the propagation of the disease have not been well understood. Akanda et al. show that cholera outbreaks in the Bengal Delta region propagate from the coast to inland and from spring to fall following two distinct transmission cycles. The first outbreak begins in the spring near the coast when northward movement of plankton-rich seawater and increasing salinity promote the growth of cholera-causing bacteria in rivers, which are used for irrigation, sanitation, and consumption. The second outbreak begins in the fall, after summer floods and monsoons affect sanitation conditions that aid in bacterial transmission by contaminating waters over much of Bangladesh. (Water Resources Research, doi:10.1029/ 2010WR009914, 2011)

  19. Ultraviolet inactivation and photoreactivation of the cholera phage 'Kappa'

    International Nuclear Information System (INIS)

    Samad, S.A.; Bhattacharyya, S.C.; Chatterjee, S.N.

    1987-01-01

    The lysogenic cholera phage, 'Kappa' is some ten to twenty folds more resistant to UV (254 nm) than are most of the T. phages of E. coli, or the cholera phage PL 163/10, or the host V. cholerae strain H218 Sm r , the 37% (D 37 ) and 10% (D 10 ) survival doses being 255.8 J/m 2 and 633.6 J/m 2 respectively. The UV-irradiated 'Kappa' phages could be photoreactivated in the host V. cholerae strain H218 Sm r to a maximum extent of 40%. The removal of the number of lethal hits per phage by the survival-enhancement treatment (photoreactivation) with time followed an exponential relation, the constant probability of removal of lethal hit per unit time being 2.8x10 -2 min -1 . The UV-irradiated phages could also be Weigle reactivated in the host strain of H218 Sm r by a small degree, the maximum reactivation factor (ratio of survivals in UV-irradiated and non-irradiated hosts) being 1.50. (orig.)

  20. Assessment of the response to cholera outbreaks in two districts in Ghana.

    Science.gov (United States)

    Ohene, Sally-Ann; Klenyuie, Wisdom; Sarpeh, Mark

    2016-11-02

    Despite recurring outbreaks of cholera in Ghana, very little has been reported on assessments of outbreak response activities undertaken in affected areas. This study assessed the response activities undertaken in two districts, Akatsi District in Volta Region and Komenda-Edina-Eguafo-Abirem (KEEA) Municipal in Central Region during the 2012 cholera epidemic in Ghana. We conducted a retrospective assessment of the events, strengths and weaknesses of the cholera outbreak response activities in the two districts making use of the WHO cholera evaluation tool. Information sources included surveillance and facility records, reports and interviews with relevant health personnel involved in the outbreak response from both district health directorates and health facilities. We collected data on age, sex, area of residence, date of reporting to health facility of cholera cases, district population data and information on the outbreak response activities and performed descriptive analyses of the outbreak data by person, time and place. The cholera outbreak in Akatsi was explosive with a high attack rate (AR) of 374/100,000 and case fatality rate (CFR) of 1.2 % while that in KEEA was on a relatively smaller scale AR of 23/100,000 but with a high case fatality rate of 18.8 %. For both districts, we identified multiple strengths in the response to the outbreak including timely notification of the district health officials which triggered prompt investigation of the suspected outbreak facilitating confirmation of cholera and initiation of public health response activities. Others were coordination of the activities by multi-sectoral committees, instituting water, sanitation and hygiene measures and appropriate case management at health facilities. We also found areas that needed improvement in both districts including incomplete surveillance data, sub-optimal community based surveillance considering the late reporting and the deaths in the community and the inadequate

  1. A cholera outbreak in Alborz Province, Iran: a matched case-control study.

    Science.gov (United States)

    Moradi, Ghobad; Rasouli, Mohammad Aziz; Mohammadi, Parvin; Elahi, Elham; Barati, Hojatollah

    2016-01-01

    A total of 229 confirmed cholera cases were reported in Alborz Province during an outbreak that lasted from June 2011 to August 2011. This study aimed to identify potential sources of transmission in order to determine suitable interventions in similar outbreaks. In other words, the lessons learned from this retrospective study can be utilized to manage future similar outbreaks. An age-matched and sex-matched case-control study was conducted during the outbreak. For each case, two control subjects were selected from the neighborhood. A case of cholera was defined as a bacteriologically confirmed case with signs and symptoms of cholera. This study was conducted from June 14, 2011 through August 23, 2011. The data were analyzed by calculating odds ratios (ORs) using the logistic regression method. In this outbreak, 229 confirmed cholera cases were diagnosed. The following risk factors were found to be associated with cholera: consumption of unrefrigerated leftover food (OR, 3.05; 95% confidence interval [CI], 1.72 to 5.41), consumption of vegetables and fruits in the previous three days (OR, 2.75; 95% CI, 1.95 to 3.89), and a history of traveling in the previous five days (OR, 5.31; 95% CI, 2.21 to 9.72). Consumption of vegetables and fruits has remained an unresolved risk factor in cholera outbreaks in Iran in recent years. In order to reduce the risk of cholera, sanitary standards for fruits and vegetables should be observed at all points from production to consumption, the population should be educated regarding hygienic food storage during outbreaks, and sanitary standards should be maintained when traveling during cholera outbreaks.

  2. A cholera outbreak in Alborz Province, Iran: a matched case-control study

    Directory of Open Access Journals (Sweden)

    Ghobad Moradi

    2016-05-01

    Full Text Available OBJECTIVES: A total of 229 confirmed cholera cases were reported in Alborz Province during an outbreak that lasted from June 2011 to August 2011. This study aimed to identify potential sources of transmission in order to determine suitable interventions in similar outbreaks. In other words, the lessons learned from this retrospective study can be utilized to manage future similar outbreaks. METHODS: An age-matched and sex-matched case-control study was conducted during the outbreak. For each case, two control subjects were selected from the neighborhood. A case of cholera was defined as a bacteriologically confirmed case with signs and symptoms of cholera. This study was conducted from June 14, 2011 through August 23, 2011. The data were analyzed by calculating odds ratios (ORs using the logistic regression method. RESULTS: In this outbreak, 229 confirmed cholera cases were diagnosed. The following risk factors were found to be associated with cholera: consumption of unrefrigerated leftover food (OR, 3.05; 95% confidence interval [CI], 1.72 to 5.41, consumption of vegetables and fruits in the previous three days (OR, 2.75; 95% CI, 1.95 to 3.89, and a history of traveling in the previous five days (OR, 5.31; 95% CI, 2.21 to 9.72. CONCLUSIONS: Consumption of vegetables and fruits has remained an unresolved risk factor in cholera outbreaks in Iran in recent years. In order to reduce the risk of cholera, sanitary standards for fruits and vegetables should be observed at all points from production to consumption, the population should be educated regarding hygienic food storage during outbreaks, and sanitary standards should be maintained when traveling during cholera outbreaks.

  3. 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; pcholera 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%; pcholera in the autumn by 29% [CI:22-33%; pcholera 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.

  4. Cholera Incidence and Mortality in Sub-Saharan African Sites during Multi-country Surveillance

    Science.gov (United States)

    Sauvageot, Delphine; Njanpop-Lafourcade, Berthe-Marie; Akilimali, Laurent; Anne, Jean-Claude; Bidjada, Pawou; Bompangue, Didier; Bwire, Godfrey; Coulibaly, Daouda; Dengo-Baloi, Liliana; Dosso, Mireille; Orach, Christopher Garimoi; Inguane, Dorteia; Kagirita, Atek; Kacou-N’Douba, Adele; Keita, Sakoba; Kere Banla, Abiba; Kouame, Yao Jean-Pierre; Landoh, Dadja Essoya; Langa, Jose Paulo; Makumbi, Issa; Miwanda, Berthe; Malimbo, Muggaga; Mutombo, Guy; Mutombo, Annie; NGuetta, Emilienne Niamke; Saliou, Mamadou; Sarr, Veronique; Senga, Raphael Kakongo; Sory, Fode; Sema, Cynthia; Tante, Ouyi Valentin; Gessner, Bradford D.; Mengel, Martin A.

    2016-01-01

    Background Cholera burden in Africa remains unknown, often because of weak national surveillance systems. We analyzed data from the African Cholera Surveillance Network (www.africhol.org). Methods/ Principal findings During June 2011–December 2013, we conducted enhanced surveillance in seven zones and four outbreak sites in Togo, the Democratic Republic of Congo (DRC), Guinea, Uganda, Mozambique and Cote d’Ivoire. All health facilities treating cholera cases were included. Cholera incidences were calculated using culture-confirmed cholera cases and culture-confirmed cholera cases corrected for lack of culture testing usually due to overwhelmed health systems and imperfect test sensitivity. Of 13,377 reported suspected cases, 34% occurred in Conakry, Guinea, 47% in Goma, DRC, and 19% in the remaining sites. From 0–40% of suspected cases were aged under five years and from 0.3–86% had rice water stools. Within surveillance zones, 0–37% of suspected cases had confirmed cholera compared to 27–38% during outbreaks. Annual confirmed incidence per 10,000 population was Conakry had corrected incidences of 20.2 and 5.8 respectively, while the other zones a median of 0.3. During outbreaks, corrected incidence varied from 2.6 to 13.0. Case fatality ratios ranged from 0–10% (median, 1%) by country. Conclusions/Significance Across different African epidemiological contexts, substantial variation occurred in cholera incidence, age distribution, clinical presentation, culture confirmation, and testing frequency. These results can help guide preventive activities, including vaccine use. PMID:27186885

  5. Cholera: tackling the epidemic in a hostile environment

    Directory of Open Access Journals (Sweden)

    Carlota GONZÁLEZ PÓZEGA

    2015-07-01

    Full Text Available The Painted Veil (2006 is a love story which takes place in the nineteen twenties, between an Englishman, Dr Walter Fane, a bacteriologist, and Kitty, an English upper class girl, who marry for convenience hardly knowing each other. Soon after the wedding they move to Shangai, where Walter is in charge of a government laboratory. Also in Shangai Kitty meets Charles, a married vice consul with whom she has an affair. When Walter discovers that his wife has been unfaithful to him he threatens to divorce her if she refuses to go with him to a village in Inner China, where there is an outbreak of cholera and where his help is required. They settle on the outskirts of Mei?tan?fu. The relationship between Walter and kitty cannot go worse; they hardly talk to each other, and while Walter is working day and night, trying to stop the spread of cholera, Kitty feels lonely and useless . One day she visits a group of French nuns who collaborate in the hospital and run an orphanage, in which Kitty is finally able to help as a music teacher. The fight against cholera is arduous: customs, religion, and politics make the doctor’s work even harder. It is then that Walter and Kitty discover qualities in each other that they did not know of; finally love and reconciliation emerge. When it seems that the outbreak of cholera is already under control, people from downstream villages, where there is no doctor, arrive. Walter feels obliged to set up a refugee camp on the outskirts of the city, where he finally contracts cholera and dies.

  6. Hurricanes, climate change and the cholera epidemic in Puerto Rico of 1855-1856.

    Science.gov (United States)

    Christenson, Bernard

    2008-01-01

    Hurricanes and global climate changes may affect the environmental factors of cholera dynamics in warm coastal areas, vulnerable to seasonal or sporadic outbreaks. The cholera epidemic of Puerto Rico in 1855-1856 had a profound effect on the Puerto Rican society; but it was not influenced by any climatic events, such as preceding hurricanes or storms based on past documentary sources. Particularly, the environmental non-toxigenic strains of Vibrio Cholerae in Puerto Rican water sources can maintain their pathogenic potential for sporadic or erratic toxigenic cholera outbreaks--if a "perfect storm" ever occurs.

  7. Natural Disasters and Cholera Outbreaks: Current Understanding and Future Outlook.

    Science.gov (United States)

    Jutla, Antarpreet; Khan, Rakibul; Colwell, Rita

    2017-03-01

    Diarrheal diseases remain a serious global public health threat, especially for those populations lacking access to safe water and sanitation infrastructure. Although association of several diarrheal diseases, e.g., cholera, shigellosis, etc., with climatic processes has been documented, the global human population remains at heightened risk of outbreak of diseases after natural disasters, such as earthquakes, floods, or droughts. In this review, cholera was selected as a signature diarrheal disease and the role of natural disasters in triggering and transmitting cholera was analyzed. Key observations include identification of an inherent feedback loop that includes societal structure, prevailing climatic processes, and spatio-temporal seasonal variability of natural disasters. Data obtained from satellite-based remote sensing are concluded to have application, although limited, in predicting risks of a cholera outbreak(s). We argue that with the advent of new high spectral and spatial resolution data, earth observation systems should be seamlessly integrated in a decision support mechanism to be mobilize resources when a region suffers a natural disaster. A framework is proposed that can be used to assess the impact of natural disasters with response to outbreak of cholera, providing assessment of short- and long-term influence of climatic processes on disease outbreaks.

  8. Spatiotemporal Variation in Environmental Vibrio cholerae in an Estuary in Southern Coastal Ecuador.

    Science.gov (United States)

    Ryan, Sadie J; Stewart-Ibarra, Anna M; Ordóñez-Enireb, Eunice; Chu, Winnie; Finkelstein, Julia L; King, Christine A; Escobar, Luis E; Lupone, Christina; Heras, Froilan; Tauzer, Erica; Waggoner, Egan; James, Tyler G; Cárdenas, Washington B; Polhemus, Mark

    2018-03-10

    Cholera emergence is strongly linked to local environmental and ecological context. The 1991-2004 pandemic emerged in Perú and spread north into Ecuador's El Oro province, making this a key site for potential re-emergence. Machala, El Oro, is a port city of 250,000 inhabitants, near the Peruvian border. Many livelihoods depend on the estuarine system, from fishing for subsistence and trade, to domestic water use. In 2014, we conducted biweekly sampling for 10 months in five estuarine locations, across a gradient of human use, and ranging from inland to ocean. We measured water-specific environmental variables implicated in cholera growth and persistence: pH, temperature, salinity, and algal concentration, and evaluated samples in five months for pathogenic and non-pathogenic Vibrio cholerae , by polymerase chain reaction (PCR). We found environmental persistence of pandemic strains O1 and O139, but no evidence for toxigenic strains. Vibrio cholerae presence was coupled to algal and salinity concentration, and sites exhibited considerable seasonal and spatial heterogeneity. This study indicates that environmental conditions in Machala are optimal for cholera re-emergence, with risk peaking during September, and higher risk near urban periphery low-income communities. This highlights a need for surveillance of this coupled cholera-estuarine system to anticipate potential future cholera outbreaks.

  9. Randomized Controlled Trial of Hospital-Based Hygiene and Water Treatment Intervention (CHoBI7) to Reduce Cholera.

    Science.gov (United States)

    George, Christine Marie; Monira, Shirajum; Sack, David A; Rashid, Mahamud-ur; Saif-Ur-Rahman, K M; Mahmud, Toslim; Rahman, Zillur; Mustafiz, Munshi; Bhuyian, Sazzadul Islam; Winch, Peter J; Leontsini, Elli; Perin, Jamie; Begum, Farzana; Zohura, Fatema; Biswas, Shwapon; Parvin, Tahmina; Zhang, Xiaotong; Jung, Danielle; Sack, R Bradley; Alam, Munirul

    2016-02-01

    The risk for cholera infection is >100 times higher for household contacts of cholera patients during the week after the index patient seeks hospital care than it is for the general population. To initiate a standard of care for this high-risk population, we developed Cholera-Hospital-Based-Intervention-for-7-Days (CHoBI7), which promotes hand washing with soap and treatment of water. To test CHoBI7, we conducted a randomized controlled trial among 219 intervention household contacts of 82 cholera patients and 220 control contacts of 83 cholera patients in Dhaka, Bangladesh, during 2013-2014. Intervention contacts had significantly fewer symptomatic Vibrio cholerae infections than did control contacts and 47% fewer overall V. cholerae infections. Intervention households had no stored drinking water with V. cholerae and 14 times higher odds of hand washing with soap at key events during structured observation on surveillance days 5, 6, or 7. CHoBI7 presents a promising approach for controlling cholera among highly susceptible household contacts of cholera patients.

  10. Avian cholera in waterfowl: the role of lesser snow and Ross's geese as carriers of avian cholera in the Playa Lakes region

    Science.gov (United States)

    Samuel, M.D.; Shadduck, D.J.; Goldberg, Diana R.; Johnson, W.P.

    2005-01-01

    We collected samples from apparently healthy geese in the Playa Lakes Region (USA) during the winters of 2000a??01 and 2001a??02 to determine whether carriers of Pasteurella multocida, the bacterium that causes avian cholera, were present in wild populations. With the use of methods developed in laboratory challenge trials (Samuel et al., 2003a) and a serotype-specific polymerase chain reaction method for identification of P. multocida serotype 1, we found that a small proportion of 322 wild birds (cholera infection. Our results confirm the hypothesis that wild waterfowl are carriers of avian cholera and add support for the hypothesis that wild birds are a reservoir for this disease. In concert with other research, this work indicates that enzootic infection with avian cholera occurs in lesser snow goose (Chen caerulescens caerulescens) populations throughout their annual cycle. Although fewer Rossa??s geese (Chen rossii) were sampled, we also found these birds were carriers of P. multocida. Even in the absence of disease outbreaks, serologic evidence indicates that chronic disease transmission and recent infection are apparently occurring year-round in these highly gregarious birds and that a small portion of these populations are potential carriers with active infection.

  11. 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.

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

    Science.gov (United States)

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

    2003-06-01

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

  13. A prolonged, community-wide cholera outbreak associated with drinking water contaminated by sewage in Kasese District, western Uganda

    Directory of Open Access Journals (Sweden)

    Benon Kwesiga

    2017-07-01

    Full Text Available Abstract Background In May 2015, a cholera outbreak that had lasted 3 months and infected over 100 people was reported in Kasese District, Uganda, where multiple cholera outbreaks had occurred previously. We conducted an investigation to identify the mode of transmission to guide control measures. Methods We defined a suspected case as onset of acute watery diarrhoea from 1 February 2015 onwards in a Kasese resident. A confirmed case was a suspected case with Vibrio cholerae O1 El Tor, serotype Inaba cultured from a stool sample. We reviewed medical records to find cases. We conducted a case-control study to compare exposures among confirmed case-persons and asymptomatic controls, matched by village and age-group. We conducted environmental assessments. We tested water samples from the most affected area for total coliforms using the Most Probable Number (MPN method. Results We identified 183 suspected cases including 61 confirmed cases of Vibrio cholerae 01; serotype Inaba, with onset between February and July 2015. 2 case-persons died of cholera. The outbreak occurred in 80 villages and affected all age groups; the highest attack rate occurred in the 5–14 year age group (4.1/10,000. The outbreak started in Bwera Sub-County bordering the Democratic Republic of Congo and spread eastward through sustained community transmission. The first case-persons were involved in cross-border trading. The case-control study, which involved 49 confirmed cases and 201 controls, showed that 94% (46/49 of case-persons compared with 79% (160/201 of control-persons drank water without boiling or treatment (ORM-H=4.8, 95% CI: 1.3–18. Water collected from the two main sources, i.e., public pipes (consumed by 39% of case-persons and 38% of control-persons or streams (consumed by 29% of case-persons and 24% control-persons had high coliform counts, a marker of faecal contamination. Environmental assessment revealed evidence of open defecation along the streams. No

  14. Cost-of-illness of cholera to households and health facilities in rural Malawi.

    Directory of Open Access Journals (Sweden)

    Patrick G Ilboudo

    Full Text Available Cholera remains an important public health problem in many low- and middle-income countries. Vaccination has been recommended as a possible intervention for the prevention and control of cholera. Evidence, especially data on disease burden, cost-of-illness, delivery costs and cost-effectiveness to support a wider use of vaccine is still weak. This study aims at estimating the cost-of-illness of cholera to households and health facilities in Machinga and Zomba Districts, Malawi. A cross-sectional study using retrospectively collected cost data was undertaken in this investigation. One hundred patients were purposefully selected for the assessment of the household cost-of-illness and four cholera treatment centres and one health facility were selected for the assessment conducted in health facilities. Data collected for the assessment in households included direct and indirect costs borne by cholera patients and their families while only direct costs were considered for the assessment conducted in health facilities. Whenever possible, descriptive and regression analysis were used to assess difference in mean costs between groups of patients. The average costs to patients' households and health facilities for treating an episode of cholera amounted to US$65.6 and US$59.7 in 2016 for households and health facilities, respectively equivalent to international dollars (I$ 249.9 and 227.5 the same year. Costs incurred in treating a cholera episode were proportional to duration of hospital stay. Moreover, 52% of households used coping strategies to compensate for direct and indirect costs imposed by the disease. Both households and health facilities could avert significant treatment expenditures through a broader use of pre-emptive cholera vaccination. These findings have direct policy implications regarding priority investments for the prevention and control of cholera.

  15. Cost-of-illness of cholera to households and health facilities in rural Malawi.

    Science.gov (United States)

    Ilboudo, Patrick G; Huang, Xiao Xian; Ngwira, Bagrey; Mwanyungwe, Abel; Mogasale, Vittal; Mengel, Martin A; Cavailler, Philippe; Gessner, Bradford D; Le Gargasson, Jean-Bernard

    2017-01-01

    Cholera remains an important public health problem in many low- and middle-income countries. Vaccination has been recommended as a possible intervention for the prevention and control of cholera. Evidence, especially data on disease burden, cost-of-illness, delivery costs and cost-effectiveness to support a wider use of vaccine is still weak. This study aims at estimating the cost-of-illness of cholera to households and health facilities in Machinga and Zomba Districts, Malawi. A cross-sectional study using retrospectively collected cost data was undertaken in this investigation. One hundred patients were purposefully selected for the assessment of the household cost-of-illness and four cholera treatment centres and one health facility were selected for the assessment conducted in health facilities. Data collected for the assessment in households included direct and indirect costs borne by cholera patients and their families while only direct costs were considered for the assessment conducted in health facilities. Whenever possible, descriptive and regression analysis were used to assess difference in mean costs between groups of patients. The average costs to patients' households and health facilities for treating an episode of cholera amounted to US$65.6 and US$59.7 in 2016 for households and health facilities, respectively equivalent to international dollars (I$) 249.9 and 227.5 the same year. Costs incurred in treating a cholera episode were proportional to duration of hospital stay. Moreover, 52% of households used coping strategies to compensate for direct and indirect costs imposed by the disease. Both households and health facilities could avert significant treatment expenditures through a broader use of pre-emptive cholera vaccination. These findings have direct policy implications regarding priority investments for the prevention and control of cholera.

  16. Assessment of Risk of Cholera in Haiti following Hurricane Matthew.

    Science.gov (United States)

    Khan, Rakib; Anwar, Rifat; Akanda, Shafqat; McDonald, Michael D; Huq, Anwar; Jutla, Antarpreet; Colwell, Rita

    2017-09-01

    Damage to the inferior and fragile water and sanitation infrastructure of Haiti after Hurricane Matthew has created an urgent public health emergency in terms of likelihood of cholera occurring in the human population. Using satellite-derived data on precipitation, gridded air temperature, and hurricane path and with information on water and sanitation (WASH) infrastructure, we tracked changing environmental conditions conducive for growth of pathogenic vibrios. Based on these data, we predicted and validated the likelihood of cholera cases occurring past hurricane. The risk of cholera in the southwestern part of Haiti remained relatively high since November 2016 to the present. Findings of this study provide a contemporary process for monitoring ground conditions that can guide public health intervention to control cholera in human population by providing access to vaccines, safe WASH facilities. Assuming current social and behavioral patterns remain constant, it is recommended that WASH infrastructure should be improved and considered a priority especially before 2017 rainy season.

  17. Survival of Vibrio cholerae O1 on fomites

    DEFF Research Database (Denmark)

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

    2016-01-01

    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...

  18. /sup 125/I-labelled tetanus toxin as a neuronal marker in tissue cultures derived from embryonic CNS

    Energy Technology Data Exchange (ETDEWEB)

    Dimpfel, W; Neale, J H; Habermann, E [Giessen Univ. (F.R. Germany). Pharmakologisches Inst.; National Inst. of Child Health and Human Development, Bethesda, Md. (USA). Behavioural Biology Branch)

    1975-01-01

    Primary cultures derived from embryonic mouse brain and spinal cord were exposed to /sup 125/I-labelled tetanus toxin and subjected to autoradioraphy. Cells with neuronal, but not glial, morphology selectively accumulated the toxin. The distribution of the grains over these cells and their processes was not uniform, discrete processes showing heavier labelling.

  19. The Role of Socioeconomic Status in Longitudinal Trends of Cholera in Matlab, Bangladesh, 1993–2007

    Science.gov (United States)

    Root, Elisabeth Dowling; Rodd, Joshua; Yunus, Mohammad; Emch, Michael

    2013-01-01

    There has been little evidence of a decline in the global burden of cholera in recent years as the number of cholera cases reported to WHO continues to rise. Cholera remains a global threat to public health and a key indicator of lack of socioeconomic development. Overall socioeconomic development is the ultimate solution for control of cholera as evidenced in developed countries. However, most research has focused on cross-county comparisons so that the role of individual- or small area-level socioeconomic status (SES) in cholera dynamics has not been carefully studied. Reported cases of cholera in Matlab, Bangladesh have fluctuated greatly over time and epidemic outbreaks of cholera continue, most recently with the introduction of a new serotype into the region. The wealth of longitudinal data on the population of Matlab provides a unique opportunity to explore the impact of socioeconomic status and other demographic characteristics on the long-term temporal dynamics of cholera in the region. In this population-based study we examine which factors impact the initial number of cholera cases in a bari at the beginning of the 0139 epidemic and the factors impacting the number of cases over time. Cholera data were derived from the ICDDR,B health records and linked to socioeconomic and geographic data collected as part of the Matlab Health and Demographic Surveillance System. Longitudinal zero-inflated Poisson (ZIP) multilevel regression models are used to examine the impact of environmental and socio-demographic factors on cholera counts across baris. Results indicate that baris with a high socioeconomic status had lower initial rates of cholera at the beginning of the 0139 epidemic (γ01 = −0.147, p = 0.041) and a higher probability of reporting no cholera cases (α01 = 0.156, p = 0.061). Populations in baris characterized by low SES are more likely to experience higher cholera morbidity at the beginning of an epidemic than populations in high SES

  20. Cholera epidemic in Guinea-Bissau (2008: the importance of "place".

    Directory of Open Access Journals (Sweden)

    Francisco J Luquero

    Full Text Available BACKGROUND: As resources are limited when responding to cholera outbreaks, knowledge about where to orient interventions is crucial. We describe the cholera epidemic affecting Guinea-Bissau in 2008 focusing on the geographical spread in order to guide prevention and control activities. METHODOLOGY/PRINCIPAL FINDINGS: We conducted two studies: 1 a descriptive analysis of the cholera epidemic in Guinea-Bissau focusing on its geographical spread (country level and within the capital; and 2 a cross-sectional study to measure the prevalence of houses with at least one cholera case in the most affected neighbourhood of the capital (Bairro Bandim to detect clustering of households with cases (cluster analysis. All cholera cases attending the cholera treatment centres in Guinea-Bissau who fulfilled a modified World Health Organization clinical case definition during the epidemic were included in the descriptive study. For the cluster analysis, a sample of houses was selected from a satellite photo (Google Earth™; 140 houses (and the four closest houses were assessed from the 2,202 identified structures. We applied K-functions and Kernel smoothing to detect clustering. We confirmed the clustering using Kulldorff's spatial scan statistic. A total of 14,222 cases and 225 deaths were reported in the country (AR = 0.94%, CFR = 1.64%. The more affected regions were Biombo, Bijagos and Bissau (the capital. Bairro Bandim was the most affected neighborhood of the capital (AR = 4.0. We found at least one case in 22.7% of the houses (95%CI: 19.5-26.2 in this neighborhood. The cluster analysis identified two areas within Bairro Bandim at highest risk: a market and an intersection where runoff accumulates waste (p<0.001. CONCLUSIONS/SIGNIFICANCE: Our analysis allowed for the identification of the most affected regions in Guinea-Bissau during the 2008 cholera outbreak, and the most affected areas within the capital. This information was essential for making

  1. Cholera epidemic in Guinea-Bissau (2008): the importance of "place".

    Science.gov (United States)

    Luquero, Francisco J; Banga, Cunhate Na; Remartínez, Daniel; Palma, Pedro Pablo; Baron, Emanuel; Grais, Rebeca F

    2011-05-04

    As resources are limited when responding to cholera outbreaks, knowledge about where to orient interventions is crucial. We describe the cholera epidemic affecting Guinea-Bissau in 2008 focusing on the geographical spread in order to guide prevention and control activities. We conducted two studies: 1) a descriptive analysis of the cholera epidemic in Guinea-Bissau focusing on its geographical spread (country level and within the capital); and 2) a cross-sectional study to measure the prevalence of houses with at least one cholera case in the most affected neighbourhood of the capital (Bairro Bandim) to detect clustering of households with cases (cluster analysis). All cholera cases attending the cholera treatment centres in Guinea-Bissau who fulfilled a modified World Health Organization clinical case definition during the epidemic were included in the descriptive study. For the cluster analysis, a sample of houses was selected from a satellite photo (Google Earth™); 140 houses (and the four closest houses) were assessed from the 2,202 identified structures. We applied K-functions and Kernel smoothing to detect clustering. We confirmed the clustering using Kulldorff's spatial scan statistic. A total of 14,222 cases and 225 deaths were reported in the country (AR = 0.94%, CFR = 1.64%). The more affected regions were Biombo, Bijagos and Bissau (the capital). Bairro Bandim was the most affected neighborhood of the capital (AR = 4.0). We found at least one case in 22.7% of the houses (95%CI: 19.5-26.2) in this neighborhood. The cluster analysis identified two areas within Bairro Bandim at highest risk: a market and an intersection where runoff accumulates waste (p<0.001). Our analysis allowed for the identification of the most affected regions in Guinea-Bissau during the 2008 cholera outbreak, and the most affected areas within the capital. This information was essential for making decisions on where to reinforce treatment and to guide control and prevention

  2. Cholera epidemics, war and disasters around Goma and Lake Kivu: an eight-year survey.

    Directory of Open Access Journals (Sweden)

    Didier Bompangue

    Full Text Available BACKGROUND: During the last eight years, North and South Kivu, located in a lake area in Eastern Democratic Republic of Congo, have been the site of a major volcano eruption and of numerous complex emergencies with population displacements. These conditions have been suspected to favour emergence and spread of cholera epidemics. METHODOLOGY/PRINCIPAL FINDINGS: In order to assess the influence of these conditions on outbreaks, reports of cholera cases were collected weekly from each health district of North Kivu (4,667,699 inhabitants and South Kivu (4,670,121 inhabitants from 2000 through 2007. A geographic information system was established, and in each health district, the relationships between environmental variables and the number of cholera cases were assessed using regression techniques and time series analysis. We further checked for a link between complex emergencies and cholera outbreaks. Finally, we analysed data collected during an epidemiological survey that was implemented in Goma after Nyiragongo eruption. A total of 73,605 cases and 1,612 deaths of cholera were reported. Time series decomposition showed a greater number of cases during the rainy season in South Kivu but not in North Kivu. Spatial distribution of cholera cases exhibited a higher number of cases in health districts bordering lakes (Odds Ratio 7.0, Confidence Interval range 3.8-12.9. Four epidemic reactivations were observed in the 12-week periods following war events, but simulations indicate that the number of reactivations was not larger than that expected during any random selection of period with no war. Nyiragongo volcanic eruption was followed by a marked decrease of cholera incidence. CONCLUSION/SIGNIFICANCE: Our study points out the crucial role of some towns located in lakeside areas in the persistence of cholera in Kivu. Even if complex emergencies were not systematically followed by cholera epidemics, some of them enabled cholera spreading.

  3. Requirement for Vibrio cholerae Integration Host Factor in Conjugative DNA Transfer

    OpenAIRE

    McLeod, Sarah M.; Burrus, Vincent; Waldor, Matthew K.

    2006-01-01

    The requirement for host factors in the transmission of integrative and conjugative elements (ICEs) has not been extensively explored. Here we tested whether integration host factor (IHF) or Fis, two host-encoded nucleoid proteins, are required for transfer of SXT, a Vibrio cholerae-derived ICE that can be transmitted to many gram-negative species. Fis did not influence the transfer of SXT to or from V. cholerae. In contrast, IHF proved to be required for V. cholerae to act as an SXT donor. I...

  4. Cholera Incidence and Mortality in Sub-Saharan African Sites during Multi-country Surveillance.

    Directory of Open Access Journals (Sweden)

    Delphine Sauvageot

    2016-05-01

    Full Text Available Cholera burden in Africa remains unknown, often because of weak national surveillance systems. We analyzed data from the African Cholera Surveillance Network (www.africhol.org.During June 2011-December 2013, we conducted enhanced surveillance in seven zones and four outbreak sites in Togo, the Democratic Republic of Congo (DRC, Guinea, Uganda, Mozambique and Cote d'Ivoire. All health facilities treating cholera cases were included. Cholera incidences were calculated using culture-confirmed cholera cases and culture-confirmed cholera cases corrected for lack of culture testing usually due to overwhelmed health systems and imperfect test sensitivity. Of 13,377 reported suspected cases, 34% occurred in Conakry, Guinea, 47% in Goma, DRC, and 19% in the remaining sites. From 0-40% of suspected cases were aged under five years and from 0.3-86% had rice water stools. Within surveillance zones, 0-37% of suspected cases had confirmed cholera compared to 27-38% during outbreaks. Annual confirmed incidence per 10,000 population was <0.5 in surveillance zones, except Goma where it was 4.6. Goma and Conakry had corrected incidences of 20.2 and 5.8 respectively, while the other zones a median of 0.3. During outbreaks, corrected incidence varied from 2.6 to 13.0. Case fatality ratios ranged from 0-10% (median, 1% by country.Across different African epidemiological contexts, substantial variation occurred in cholera incidence, age distribution, clinical presentation, culture confirmation, and testing frequency. These results can help guide preventive activities, including vaccine use.

  5. Risk factors associated with cholera in Harare City, Zimbabwe, 2008 ...

    African Journals Online (AJOL)

    Objective: Two suspected cholera cases at Beatrice Road Infectious Diseases Hospital were reported to Harare City Health on 14 October 2008 setting in motion investigation and control measures. We determined the extent of the epidemic and risk factors for contracting cholera. Methods: An unmatched 1:1 case-control ...

  6. Expression of cholera toxin B–proinsulin fusion protein in lettuce and tobacco chloroplasts – oral administration protects against development of insulitis in non-obese diabetic mice

    Science.gov (United States)

    Ruhlman, Tracey; Ahangari, Raheleh; Devine, Andrew; Samsam, Mohtahsem; Daniell, Henry

    2008-01-01

    Summary Lettuce and tobacco chloroplast transgenic lines expressing the cholera toxin B subunit–human proinsulin (CTB-Pins) fusion protein were generated. CTB-Pins accumulated up to ~16% of total soluble protein (TSP) in tobacco and up to ~2.5% of TSP in lettuce. Eight milligrams of powdered tobacco leaf material expressing CTB-Pins or, as negative controls, CTB–green fluorescent protein (CTB-GFP) or interferon–GFP (IFN-GFP), or untransformed leaf, were administered orally, each week for 7 weeks, to 5-week-old female non-obese diabetic (NOD) mice. The pancreas of CTB-Pins-treated mice showed decreased infiltration of cells characteristic of lymphocytes (insulitis); insulin-producing β-cells in the pancreatic islets of CTB-Pins-treated mice were significantly preserved, with lower blood or urine glucose levels, by contrast with the few β-cells remaining in the pancreatic islets of the negative controls. Increased expression of immunosuppressive cytokines, such as interleukin-4 and interleukin-10 (IL-4 and IL-10), was observed in the pancreas of CTB-Pins-treated NOD mice. Serum levels of immunoglobulin G1 (IgG1), but not IgG2a, were elevated in CTB-Pins-treated mice. Taken together, T-helper 2 (Th2) lymphocyte-mediated oral tolerance is a likely mechanism for the prevention of pancreatic insulitis and the preservation of insulin-producing β-cells. This is the first report of expression of a therapeutic protein in transgenic chloroplasts of an edible crop. Transplastomic lettuce plants expressing CTB-Pins grew normally and transgenes were maternally inherited in T1 progeny. This opens up the possibility for the low-cost production and delivery of human therapeutic proteins, and a strategy for the treatment of various other autoimmune diseases. PMID:17490448

  7. DPN-Generated Combinatorial Libraries

    Science.gov (United States)

    2012-02-29

    µM final concentration) is added (Fig. 6). Control over deposition parameters was examined for two model proteins, cholera toxin β subunit (CTβ...conjugated anti- cholera toxin beta (anti-CTb). The wells in the mould are inverted pyramids with an average depth of 86 µm, edge length of 120 µm, and...Therapeutics,” (2011). 91. University of New Mexico , Chemistry Department Colloquium, Albuquerque, NM, “The Polyvalent Gold Nanoparticle Conjugate

  8. Rainfall mediations in the spreading of epidemic cholera

    Science.gov (United States)

    Righetto, L.; Bertuzzo, E.; Mari, L.; Schild, E.; Casagrandi, R.; Gatto, M.; Rodriguez-Iturbe, I.; Rinaldo, A.

    2013-10-01

    Following the empirical evidence of a clear correlation between rainfall events and cholera resurgence that was observed in particular during the recent outbreak in Haiti, a spatially explicit model of epidemic cholera is re-examined. Specifically, we test a multivariate Poisson rainfall generator, with parameters varying in space and time, as a driver of enhanced disease transmission. The relevance of the issue relates to the key insight that predictive mathematical models may provide into the course of an ongoing cholera epidemic aiding emergency management (say, in allocating life-saving supplies or health care staff) or in evaluating alternative management strategies. Our model consists of a set of dynamical equations (SIRB-like i.e. subdivided into the compartments of Susceptible, Infected and Recovered individuals, and including a balance of Bacterial concentrations in the water reservoir) describing a connected network of human communities where the infection results from the exposure to excess concentrations of pathogens in the water. These, in turn, are driven by rainfall washout of open-air defecation sites or cesspool overflows, hydrologic transport through waterways and by mobility of susceptible and infected individuals. We perform an a posteriori analysis (from the beginning of the epidemic in October 2010 until December 2011) to test the model reliability in predicting cholera cases and in testing control measures, involving vaccination and sanitation campaigns, for the ongoing epidemic. Even though predicting reliably the timing of the epidemic resurgence proves difficult due to rainfall inter-annual variability, we find that the model can reasonably quantify the total number of reported infection cases in the selected time-span. We then run a multi-seasonal prediction of the course of the epidemic until December 2015, to investigate conditions for further resurgences and endemicity of cholera in the region with a view to policies which may bring to

  9. Vibrio cholerae classical biotype strains reveal distinct signatures in Mexico.

    Science.gov (United States)

    Alam, Munirul; Islam, M Tarequl; Rashed, Shah Manzur; Johura, Fatema-tuz; Bhuiyan, Nurul A; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Watanabe, Haruo; Hasan, Nur-A; Colwell, Rita R; Cravioto, Alejandro

    2012-07-01

    Vibrio cholerae O1 classical (CL) biotype caused the fifth and sixth pandemics, and probably the earlier cholera pandemics, before the El Tor (ET) biotype initiated the seventh pandemic in Asia in the 1970s by completely displacing the CL biotype. Although the CL biotype was thought to be extinct in Asia and although it had never been reported from Latin America, V. cholerae CL and ET biotypes, including a hybrid ET, were found associated with areas of cholera endemicity in Mexico between 1991 and 1997. In this study, CL biotype strains isolated from areas of cholera endemicity in Mexico between 1983 and 1997 were characterized in terms of major phenotypic and genetic traits and compared with CL biotype strains isolated in Bangladesh between 1962 and 1989. According to sero- and biotyping data, all V. cholerae strains tested had the major phenotypic and genotypic characteristics specific for the CL biotype. Antibiograms revealed the majority of the Bangladeshi strains to be resistant to trimethoprim-sulfamethoxazole, furazolidone, ampicillin, and gentamicin, while the Mexican strains were sensitive to all of these drugs, as well as to ciprofloxacin, erythromycin, and tetracycline. Pulsed-field gel electrophoresis (PFGE) of NotI-digested genomic DNA revealed characteristic banding patterns for all of the CL biotype strains although the Mexican strains differed from the Bangladeshi strains in 1 to 2 DNA bands. The difference was subtle but consistent, as confirmed by the subclustering patterns in the PFGE-based dendrogram, and can serve as a regional signature, suggesting the pre-1991 existence and evolution of the CL biotype strains in the Americas, independent from Asia.

  10. Laboratory and Clinical features of EIA Toxin-positive and EIA Toxin-negative Community-acquired Clostridium difficile Infection.

    Science.gov (United States)

    Patel, Hiren; Randhawa, Jeewanjot; Nanavati, Sushant; Marton, L Randy; Baddoura, Walid J; DeBari, Vincent A

    2015-01-01

    Studies have described the clinical course of patients with Clostridium difficile infection (CDI) with positive enzyme immunoassay (EIA) for toxins A and B. Limited information is available for the patients with negative EIA but positive for the toxin B gene (TcdB) by the PCR. The aim of our study is to determine if there are any differences that exist among the clinical and laboratory parameters in the patients tested to be positive by EIA for toxin and those who were negative. This is a retrospective cohort study conducted in a 700-bed teaching hospital. We reviewed charts of the patients with presumptive CDI between January 2006 and July 2013. We divided these patients into two groups, EIA-positive and EIA-negative, based on result of EIA for toxins A and B and the requirement for a positive PCR analysis of the TcdB gene. The EIA-positive group had significantly higher white blood cell counts (p<0.001), with a significantly greater percentage of bands (p<0.0001). Albumin and total protein both exhibit significantly (p<0.0001, both comparisons) lower values in the EIA-positive group. Among clinical findings, the EIA-positive group had significantly longer length of hospital stay (p=0.010). These data suggest that an infection with an EIA-negative strain of C. difficile presents laboratory markers closer to those of healthy subjects and clinical features suggesting considerably less severe than infection with EIA-positive C. difficile. © 2015 by the Association of Clinical Scientists, Inc.

  11. Informal urban settlements and cholera risk in Dar es Salaam, Tanzania.

    Directory of Open Access Journals (Sweden)

    Katherine Penrose

    2010-03-01

    Full Text Available As a result of poor economic opportunities and an increasing shortage of affordable housing, much of the spatial growth in many of the world's fastest-growing cities is a result of the expansion of informal settlements where residents live without security of tenure and with limited access to basic infrastructure. Although inadequate water and sanitation facilities, crowding and other poor living conditions can have a significant impact on the spread of infectious diseases, analyses relating these diseases to ongoing global urbanization, especially at the neighborhood and household level in informal settlements, have been infrequent. To begin to address this deficiency, we analyzed urban environmental data and the burden of cholera in Dar es Salaam, Tanzania.Cholera incidence was examined in relation to the percentage of a ward's residents who were informal, the percentage of a ward's informal residents without an improved water source, the percentage of a ward's informal residents without improved sanitation, distance to the nearest cholera treatment facility, population density, median asset index score in informal areas, and presence or absence of major roads. We found that cholera incidence was most closely associated with informal housing, population density, and the income level of informal residents. Using data available in this study, our model would suggest nearly a one percent increase in cholera incidence for every percentage point increase in informal residents, approximately a two percent increase in cholera incidence for every increase in population density of 1000 people per km(2 in Dar es Salaam in 2006, and close to a fifty percent decrease in cholera incidence in wards where informal residents had minimally improved income levels, as measured by ownership of a radio or CD player on average, in comparison to wards where informal residents did not own any items about which they were asked. In this study, the range of access to

  12. Informal Urban Settlements and Cholera Risk in Dar es Salaam, Tanzania

    Science.gov (United States)

    Penrose, Katherine; de Castro, Marcia Caldas; Werema, Japhet; Ryan, Edward T.

    2010-01-01

    Background As a result of poor economic opportunities and an increasing shortage of affordable housing, much of the spatial growth in many of the world's fastest-growing cities is a result of the expansion of informal settlements where residents live without security of tenure and with limited access to basic infrastructure. Although inadequate water and sanitation facilities, crowding and other poor living conditions can have a significant impact on the spread of infectious diseases, analyses relating these diseases to ongoing global urbanization, especially at the neighborhood and household level in informal settlements, have been infrequent. To begin to address this deficiency, we analyzed urban environmental data and the burden of cholera in Dar es Salaam, Tanzania. Methodology/Principal Findings Cholera incidence was examined in relation to the percentage of a ward's residents who were informal, the percentage of a ward's informal residents without an improved water source, the percentage of a ward's informal residents without improved sanitation, distance to the nearest cholera treatment facility, population density, median asset index score in informal areas, and presence or absence of major roads. We found that cholera incidence was most closely associated with informal housing, population density, and the income level of informal residents. Using data available in this study, our model would suggest nearly a one percent increase in cholera incidence for every percentage point increase in informal residents, approximately a two percent increase in cholera incidence for every increase in population density of 1000 people per km2 in Dar es Salaam in 2006, and close to a fifty percent decrease in cholera incidence in wards where informal residents had minimally improved income levels, as measured by ownership of a radio or CD player on average, in comparison to wards where informal residents did not own any items about which they were asked. In this study, the

  13. Informal urban settlements and cholera risk in Dar es Salaam, Tanzania.

    Science.gov (United States)

    Penrose, Katherine; de Castro, Marcia Caldas; Werema, Japhet; Ryan, Edward T

    2010-03-16

    As a result of poor economic opportunities and an increasing shortage of affordable housing, much of the spatial growth in many of the world's fastest-growing cities is a result of the expansion of informal settlements where residents live without security of tenure and with limited access to basic infrastructure. Although inadequate water and sanitation facilities, crowding and other poor living conditions can have a significant impact on the spread of infectious diseases, analyses relating these diseases to ongoing global urbanization, especially at the neighborhood and household level in informal settlements, have been infrequent. To begin to address this deficiency, we analyzed urban environmental data and the burden of cholera in Dar es Salaam, Tanzania. Cholera incidence was examined in relation to the percentage of a ward's residents who were informal, the percentage of a ward's informal residents without an improved water source, the percentage of a ward's informal residents without improved sanitation, distance to the nearest cholera treatment facility, population density, median asset index score in informal areas, and presence or absence of major roads. We found that cholera incidence was most closely associated with informal housing, population density, and the income level of informal residents. Using data available in this study, our model would suggest nearly a one percent increase in cholera incidence for every percentage point increase in informal residents, approximately a two percent increase in cholera incidence for every increase in population density of 1000 people per km(2) in Dar es Salaam in 2006, and close to a fifty percent decrease in cholera incidence in wards where informal residents had minimally improved income levels, as measured by ownership of a radio or CD player on average, in comparison to wards where informal residents did not own any items about which they were asked. In this study, the range of access to improved sanitation

  14. Laboratory evaluation of immunochromatographic rapid diagnostic tests for cholera in Haiti.

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    Wilfredo R Matias

    Full Text Available Rapid diagnostic tests (RDT for cholera are promising tools for detecting cholera in areas with limited laboratory infrastructure. However, evidence on the characteristics of the many available RDTs is scarce, and their use has been limited by suboptimal performance. We evaluated the performance characteristics of three cholera RDTs from Span Diagnostics, Artron Laboratories, and Standard Diagnostics in a regional laboratory in Haiti.We retrospectively reviewed records from May 2014 to October 2015 of a laboratory-based surveillance program for Vibrio cholerae at Hôpital Saint-Nicolas in Saint-Marc, Haiti. We compared the results of 511 Crystal VC, 129 Artron and 451 SD Bioline RDTs to bacterial culture as the gold standard. Of 905 cultures, 477 (52.7% were positive for V. cholerae O1, of which 27.7% were serotype Inaba. No cultures grew V. cholerae O139. Sensitivity and specificity of Crystal VC were 98.6% (95%CI: 96.5%-99.6% and 71.1% (95%CI: 64.7%-76.9%, respectively. Artron demonstrated a sensitivity of 98.6% (95%CI: 92.7%-100% and specificity of 69.1% (95%CI: 55.2%-80.9%. SD Bioline demonstrated a sensitivity of 81.1% (95%CI: 75.6%-85.8% and specificity of 92.8% (95%CI: 88.4%-95.9%. Crystal VC and Artron frequently showed false positive O139 bands, whereas none were seen with SD Bioline.There is significant variation in the performance of different cholera diagnostic RDTs. Artron and Crystal VC RDTs have high sensitivity and low specificity, while SD Bioline RDT has low to moderate sensitivity and high specificity when performed by laboratory technicians in Haiti. Study limitations included its retrospective design. The suboptimal characteristics of these tests limit their use as clinical point-of-care tests; however, they may be useful in outbreak response, surveillance, and research in resource-limited settings.

  15. Drinking cholera: salinity levels and palatability of drinking water in coastal Bangladesh.

    Science.gov (United States)

    Grant, Stephen Lawrence; Tamason, Charlotte Crim; Hoque, Bilqis Amin; Jensen, Peter Kjaer Mackie

    2015-04-01

    To measure the salinity levels of common water sources in coastal Bangladesh and explore perceptions of water palatability among the local population to investigate the plausibility of linking cholera outbreaks in Bangladesh with ingestion of saline-rich cholera-infected river water. Hundred participants took part in a taste-testing experiment of water with varying levels of salinity. Salinity measurements were taken of both drinking and non-drinking water sources. Informal group discussions were conducted to gain an in-depth understanding of water sources and water uses. Salinity levels of non-drinking water sources suggest that the conditions for Vibrio cholerae survival exist 7-8 days within the local aquatic environment. However, 96% of participants in the taste-testing experiment reported that they would never drink water with salinity levels that would be conducive to V. cholerae survival. Furthermore, salinity levels of participant's drinking water sources were all well below the levels required for optimal survival of V. cholerae. Respondents explained that they preferred less salty and more aesthetically pleasing drinking water. Theoretically, V. cholerae can survive in the river systems in Bangladesh; however, water sources which have been contaminated with river water are avoided as potential drinking water sources. Furthermore, there are no physical connecting points between the river system and drinking water sources among the study population, indicating that the primary driver for cholera cases in Bangladesh is likely not through the contamination of saline-rich river water into drinking water sources. © 2015 John Wiley & Sons Ltd.

  16. Research Protocol - Cholera and pregnancy in Haiti: description of pregnant patients presenting to MSF OCA cholera treatment centers, September 2011-December 2013.

    OpenAIRE

    Schillberg, Erin; Bryson, Lindsay; Pierre, Grand; Lenglet, Annick

    2015-01-01

    Principal objective To understand the demographic, clinical and outcome profiles of pregnant patients that presented with cholera infection to Figaro CTC and CRUO CTU between September 2011 and December 2013. Specific objectives 1. To determine the clinical presentation, treatment regimens and outcomes of pregnant patients with cholera seen at Figaro CTC and CRUO CTU between September 2011 and December 2014; 2. To identify factors related to age, clinical presentation or treatmen...

  17. Stool C difficile toxin

    Science.gov (United States)

    ... toxin; Colitis - toxin; Pseudomembranous - toxin; Necrotizing colitis - toxin; C difficile - toxin ... be analyzed. There are several ways to detect C difficile toxin in the stool sample. Enzyme immunoassay ( ...

  18. PAR-1 mediated apoptosis of breast cancer cells by V. cholerae hemagglutinin protease.

    Science.gov (United States)

    Ray, Tanusree; Pal, Amit

    2016-05-01

    Bacterial toxins have emerged as promising agents in cancer treatment strategy. Hemagglutinin (HAP) protease secreted by Vibrio cholerae induced apoptosis in breast cancer cells and regresses tumor growth in mice model. The success of novel cancer therapies depends on their selectivity for cancer cells with limited toxicity for normal tissues. Increased expression of Protease Activated Receptor-1 (PAR-1) has been reported in different malignant cells. In this study we report that HAP induced activation and over expression of PAR-1 in breast cancer cells (EAC). Immunoprecipitation studies have shown that HAP specifically binds with PAR-1. HAP mediated activation of PAR-1 caused nuclear translocation of p50-p65 and the phosphorylation of p38 which triggered the activation of NFκB and MAP kinase signaling pathways. These signaling pathways enhanced the cellular ROS level in malignant cells that induced the intrinsic pathway of cell apoptosis. PAR-1 mediated apoptosis by HAP of malignant breast cells without effecting normal healthy cells in the same environment makes it a good therapeutic agent for treatment of cancer.

  19. Social and cultural features of cholera and shigellosis in peri-urban and rural communities of Zanzibar

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    Hutubessy Raymond

    2010-11-01

    Full Text Available Abstract Background Responding to the high burden of cholera in developing countries, the WHO now considers vaccination as a supplement to the provision of safe drinking water and improved sanitation in the strategy for cholera control in endemic settings. Cultural concepts of illness affect many aspects of public health. In the first step of a two-step strategy to examine determinants of cholera vaccine acceptance, this study identified social and cultural features of diarrhoeal illness for cholera control in endemic communities. Methods A cultural epidemiological study with locally adapted vignette-based interviews was conducted in two cholera-endemic communities of Zanzibar. A random sample of unaffected peri-urban (n = 179 and rural (n = 177 adults was interviewed to study community ideas of cholera and shigellosis, considering categories of distress, perceived causes, and help-seeking behaviour. Results Cholera was recognised by 88%. Symptoms of dehydration were most prominent in reports at the peri-urban site. Interference with work leading to strain on household finances was frequently emphasised. Dirty environment was the most prominent perceived cause, followed by unsafe drinking water and germ-carrying flies. Causes unrelated to the biomedical basis of cholera were reported more often by rural respondents. Rural women had more difficulty (20% to identify a cause than men (7.1%, p = 0.016. Peri-urban self treatment emphasised rehydration; the rural community preferred herbal treatment and antibiotics. Shigellosis was recognised by 70%. Fewer regarded it as very serious compared with cholera (76% vs. 97%, p Conclusions This study clarified local views of cholera and shigellosis relevant for diarrhoeal disease control in Zanzibar. The finding that rural women were less likely than men to specify causes of cholera suggests more attention to them is required. Better health education is needed for cholera in rural areas and for shigellosis

  20. Genetic characterization of ØVC8 lytic phage for Vibrio cholerae O1.

    Science.gov (United States)

    Solís-Sánchez, Alejandro; Hernández-Chiñas, Ulises; Navarro-Ocaña, Armando; De la Mora, Javier; Xicohtencatl-Cortes, Juan; Eslava-Campos, Carlos

    2016-03-22

    Epidemics and pandemics of cholera, a diarrheal disease, are attributed to Vibrio cholera serogroups O1 and O139. In recent years, specific lytic phages of V. cholera have been proposed to be important factors in the cyclic occurrence of cholera in endemic areas. However, the role and potential participation of lytic phages during long interepidemic periods of cholera in non-endemic regions have not yet been described. The purpose of this study was to isolate and characterize specific lytic phages of V. cholera O1 strains. Sixteen phages were isolated from wastewater samples collected at the Endhó Dam in Hidalgo State, Mexico, concentrated with PEG/NaCl, and purified by density gradient. The lytic activity of the purified phages was tested using different V. cholerae O1 and O139 strains. Phage morphology was visualized by transmission electron microscopy (TEM), and phage genome sequencing was performed using the Genome Analyzer IIx System. Genome assembly and bioinformatics analysis were performed using a set of high-throughput programs. Phage structural proteins were analyzed by mass spectrometry. Sixteen phages with lytic and lysogenic activity were isolated; only phage ØVC8 showed specific lytic activity against V. cholerae O1 strains. TEM images of ØVC8 revealed a phage with a short tail and an isometric head. The ØVC8 genome comprises linear double-stranded DNA of 39,422 bp with 50.8 % G + C. Of the 48 annotated ORFs, 16 exhibit homology with sequences of known function and several conserved domains. Bioinformatics analysis showed multiple conserved domains, including an Ig domain, suggesting that ØVC8 might adhere to different mucus substrates such as the human intestinal epithelium. The results suggest that ØVC8 genome utilize the "single-stranded cohesive ends" packaging strategy of the lambda-like group. The two structural proteins sequenced and analyzed are proteins of known function. ØVC8 is a lytic phage with specific activity against V. cholerae

  1. Hydroclimatological Controls of Endemic and Non-endemic Cholera of the 20th Century

    Science.gov (United States)

    Jutla, A. S.; Whitcombe, E.; Colwell, R.

    2012-12-01

    Cholera remains a major public health threat for the developing countries. Since the causative agent, Vibrio cholerae, is autochthonous to aquatic environment, it is not possible to eradicate the agent of the disease. Hydroclimatology based prediction and prevention strategies can be implemented in disease susceptible regions for reducing incidence rates. However, the precise role of hydrological and climatological processes, which will further aid in development of suitable prediction models, in creating spatial and temporal environmental conditions favorable for disease outbreak has not been adequately quantified. Here, we show distinction between seasonality and occurrence of cholera in epidemic and non-endemic regions. Using historical cholera mortality data, from the late 1800s for 27 locations in the Indian subcontinent, we show that non-endemic regions are generally located close to regional river systems but away from the coasts and are characterized by single sporadic outbreak in a given year. Increase in air temperature during the low river flow season increases evaporation, leading to an optimal salinity and pH required for bacterial growth. Thereafter, monsoonal rainfall, leads to interactions of contaminated river waters via human activity resulting in cholera epidemics. Endemic regions are located close to coasts where cholera outbreak occurs twice (spring and fall) in a year. Spring outbreak is generally associated with intrusion of bacterial seawater to inland whereas the fall peak is correlated with widespread flooding and cross-contamination of water resources via increased precipitation. This may be one of the first studies to hydroclimatologically quantitatively the seasonality of cholera in both endemic and non-endemic regions. Our results prompt the need of region and cause-specific prediction models for cholera, employing appropriate environmental determinants.

  2. Human resources for health: lessons from the cholera outbreak in Papua New Guinea.

    Science.gov (United States)

    Rosewell, Alexander; Bieb, Sibauk; Clark, Geoff; Miller, Geoff; MacIntyre, Raina; Zwi, Anthony

    2013-01-01

    Papua New Guinea is striving to achieve the minimum core requirements under the International Health Regulations in surveillance and outbreak response, and has experienced challenges in the availability and distribution of health professionals. Since mid-2009, a large cholera outbreak spread across lowland regions of the country and has been associated with more than 15 500 notifications at a case fatality ratio of 3.2%. The outbreak placed significant pressure on clinical and public health services. We describe some of the challenges to cholera preparedness and response in this human resource-limited setting, the strategies used to ensure effective cholera management and lessons learnt. Cholera task forces were useful to establish a clear system of leadership and accountability for cholera outbreak response and ensure efficiencies in each technical area. Cholera outbreak preparedness and response was strongest when human resource and health systems functioned well before the outbreak. Communication relied on coordination of existing networks and methods for empowering local leaders and villagers to modify behaviours of the population. In line with the national health emergencies plan, the successes of human resource strategies during the cholera outbreak should be built upon through emergency exercises, especially in non-affected provinces. Population needs for all public health professionals involved in health emergency preparedness and response should be mapped, and planning should be implemented to increase the numbers in relevant areas. Human resource planning should be integrated with health emergency planning. It is essential to maintain and strengthen the human resource capacities and experiences gained during the cholera outbreak to ensure a more effective response to the next health emergency.

  3. Estimating the cost of cholera-vaccine delivery from the societal point of view: A case of introduction of cholera vaccine in Bangladesh.

    Science.gov (United States)

    Sarker, Abdur Razzaque; Islam, Ziaul; Khan, Iqbal Ansary; Saha, Amit; Chowdhury, Fahima; Khan, Ashraful Islam; Cravioto, Alejandro; Clemens, John David; Qadri, Firdausi; Khan, Jahangir A M

    2015-09-11

    Cholera is a major global public health problem that causes both epidemic and endemic disease. The World Health Organization recommends oral cholera vaccines as a public health tool in addition to traditional prevention practices and treatments in both epidemic and endemic settings. In many developing countries like Bangladesh, the major issue concerns the affordability of this vaccine. In February 2011, a feasibility study entitled, "Introduction of Cholera Vaccine in Bangladesh (ICVB)", was conducted for a vaccination campaign using inactivated whole-cell cholera vaccine (Shanchol) in a high risk area of Mirpur, Dhaka. Empirical data obtained from this trial was used to determine the vaccination cost for a fully immunized person from the societal perspective. A total of 123,661 people were fully vaccinated receiving two doses of the vaccine, while 18,178 people received one dose of the same vaccine. The total cost for vaccine delivery was US$ 492,238 giving a total vaccination cost per fully-vaccinated individual of US$ 3.98. The purchase cost of the vaccine accounted for 58% of the overall cost of vaccination. Attempts to reduce the per-dose cost of the vaccine are likely to have a large impact on the cost of similar vaccination campaigns in the future. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. A highly specific phage defense system is a conserved feature of the Vibrio cholerae mobilome.

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    Brendan J O'Hara

    2017-06-01

    Full Text Available Vibrio cholerae-specific bacteriophages are common features of the microbial community during cholera infection in humans. Phages impose strong selective pressure that favors the expansion of phage-resistant strains over their vulnerable counterparts. The mechanisms allowing virulent V. cholerae strains to defend against the ubiquitous threat of predatory phages have not been established. Here, we show that V. cholerae PLEs (phage-inducible chromosomal island-like elements are widespread genomic islands dedicated to phage defense. Analysis of V. cholerae isolates spanning a 60-year collection period identified five unique PLEs. Remarkably, we found that all PLEs (regardless of geographic or temporal origin respond to infection by a myovirus called ICP1, the most prominent V. cholerae phage found in cholera patient stool samples from Bangladesh. We found that PLE activity reduces phage genome replication and accelerates cell lysis following ICP1 infection, killing infected host cells and preventing the production of progeny phage. PLEs are mobilized by ICP1 infection and can spread to neighboring cells such that protection from phage predation can be horizontally acquired. Our results reveal that PLEs are a persistent feature of the V. cholerae mobilome that are adapted to providing protection from a single predatory phage and advance our understanding of how phages influence pathogen evolution.

  5. A highly specific phage defense system is a conserved feature of the Vibrio cholerae mobilome.

    Science.gov (United States)

    O'Hara, Brendan J; Barth, Zachary K; McKitterick, Amelia C; Seed, Kimberley D

    2017-06-01

    Vibrio cholerae-specific bacteriophages are common features of the microbial community during cholera infection in humans. Phages impose strong selective pressure that favors the expansion of phage-resistant strains over their vulnerable counterparts. The mechanisms allowing virulent V. cholerae strains to defend against the ubiquitous threat of predatory phages have not been established. Here, we show that V. cholerae PLEs (phage-inducible chromosomal island-like elements) are widespread genomic islands dedicated to phage defense. Analysis of V. cholerae isolates spanning a 60-year collection period identified five unique PLEs. Remarkably, we found that all PLEs (regardless of geographic or temporal origin) respond to infection by a myovirus called ICP1, the most prominent V. cholerae phage found in cholera patient stool samples from Bangladesh. We found that PLE activity reduces phage genome replication and accelerates cell lysis following ICP1 infection, killing infected host cells and preventing the production of progeny phage. PLEs are mobilized by ICP1 infection and can spread to neighboring cells such that protection from phage predation can be horizontally acquired. Our results reveal that PLEs are a persistent feature of the V. cholerae mobilome that are adapted to providing protection from a single predatory phage and advance our understanding of how phages influence pathogen evolution.

  6. 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.

  7. Flow cytofluorometric monitoring of leukocyte apoptosis in experimental cholera

    Science.gov (United States)

    Lotsmanova, Ekaterina Y.; Kravtsov, Alexander L.; Livanova, Ludmila F.; Kobkova, Irina M.; Kuznetsov, Oleg S.; Shchukovskaya, Tatyana N.; Smirnova, Nina I.; Kutyrev, Vladimir V.

    2003-10-01

    Flow cytofluorometric DNA analysis was applied to determine of the relative contents of proliferative (more then 2C DNA per cell) and apoptotic (less then 2C DNA per cell) leukocytes in blood of adult rabbits, challenged with 10,000 times the 50 % effective dose of Vibrio cholerae virulent strain by the RITARD technique. It has been shown that irreversible increase the percentage of cells carrying DNA in the degradation stage brings to disbalance between the genetically controlled cell proliferation and apoptosis that leads to animal death from the cholera infection. Such fatal changes were not observed in challenging of immunized animals that were not died. Thus received data show that the flow cytofluorometric measurements may be used for detection of transgressions in homeostasis during acute infection diseases, for outlet prognosis of the cholera infection.

  8. Cyclic AMP in rat pancreatic islets

    International Nuclear Information System (INIS)

    Grill, V.; Borglund, E.; Cerasi, E.; Uppsala Univ.

    1977-01-01

    The incorporation of [ 3 H]adenine into cyclic AMP was studied in rat pancreatic islets under varying conditions of labeling. Prolonging the exposure to [ 3 H]adenine progressively augmented the islet cyclic [ 3 H]AMP level. Islets labeled for different periods of time and subsequently incubated (without adenine) in the presence of D-glucose or cholera toxin showed stimulations of intra-islet cyclic [ 3 H]AMP that were proportionate to the levels of radioactive nucleotide present under non-stimulatory conditions. Labeling the islets in a high glucose concentration (27.7 mM) did not modify the nucleotide responses to glucose or cholera toxin. The specific activity of cyclic [ 3 H]AMP, determined by simultaneous assay of cyclic [ 3 H]AMP and total cyclic AMP, was not influenced by glucose or cholera toxin. Glucose had no effect on the specific activity of labeled ATP

  9. Microbial glycolipoprotein-capped silver nanoparticles as emerging antibacterial agents against cholera.

    Science.gov (United States)

    Gahlawat, Geeta; Shikha, Sristy; Chaddha, Baldev Singh; Chaudhuri, Saumya Ray; Mayilraj, Shanmugam; Choudhury, Anirban Roy

    2016-02-01

    With the increased number of cholera outbreaks and emergence of multidrug resistance in Vibrio cholerae strains it has become necessary for the scientific community to devise and develop novel therapeutic approaches against cholera. Recent studies have indicated plausibility of therapeutic application of metal nano-materials. Among these, silver nanoparticles (AgNPs) have emerged as a potential antimicrobial agent to combat infectious diseases. At present nanoparticles are mostly produced using physical or chemical techniques which are toxic and hazardous. Thus exploitation of microbial systems could be a green eco-friendly approach for the synthesis of nanoparticles having similar or even better antimicrobial activity and biocompatibility. Hence, it would be worth to explore the possibility of utilization of microbial silver nanoparticles and their conjugates as potential novel therapeutic agent against infectious diseases like cholera. The present study attempted utilization of Ochrobactrum rhizosphaerae for the production of AgNPs and focused on investigating their role as antimicrobial agents against cholera. Later the exopolymer, purified from the culture supernatant, was used for the synthesis of spherical shaped AgNPs of around 10 nm size. Further the exopolymer was characterized as glycolipoprotein (GLP). Antibacterial activity of the novel GLP-AgNPs conjugate was evaluated by minimum inhibitory concentration, XTT reduction assay, scanning electron microscopy (SEM) and growth curve analysis. SEM studies revealed that AgNPs treatment resulted in intracellular contents leakage and cell lysis. The potential of microbially synthesized nanoparticles, as novel therapeutic agents, is still relatively less explored. In fact, the present study first time demonstrated that a glycolipoprotein secreted by the O. rhizosphaerae strain can be exploited for production of AgNPs which can further be employed to treat infectious diseases. Although this type of polymer has

  10. Clinical Features of Human Immunodeficiency Virus-Infected Patients Presenting with Cholera in Port-au-Prince, Haiti.

    Science.gov (United States)

    Sévère, Karine; Anglade, Stravinsky B; Bertil, Claudin; Duncan, Aynsley; Joseph, Patrice; Deroncenay, Alexandra; Mabou, Marie M; Ocheretina, Oksana; Reif, Lindsey; Seo, Grace; Pape, Jean W; Fitzgerald, Daniel W

    2016-11-02

    Human immunodeficiency virus (HIV) infection has been postulated to alter the natural history of cholera, including increased susceptibility to infection, severity of illness, and chronic carriage of Vibrio cholerae Haiti has a generalized HIV epidemic with an adult HIV prevalence of 1.9% and recently suffered a cholera epidemic. We conducted a prospective study at the cholera treatment center (CTC) of GHESKIO in Haiti to characterize the coinfection. Adults admitted at the CTC for acute diarrhea were invited to participate in the study. Vital signs, frequency, and volume of stools and/or vomiting were monitored, and single-dose doxycycline was administered. After counseling, participants were screened for HIV by enzyme-linked immunosorbent assay and for cholera by culture. Of 729 adults admitted to the CTC, 99 (13.6%) had HIV infection, and 457 (63%) had culture-confirmed cholera. HIV prevalence was three times higher in patients without cholera (23%, 63/272) than in those with culture-confirmed cholera (7.9%, 36/457). HIV prevalence in patients with culture-confirmed cholera (7.9%) was four times higher than the adult prevalence in Port-au-Prince (1.9%). Of the 36 HIV-infected patients with cholera, 25 (69%) had moderate/severe dehydration versus 302/421 (72%) in the HIV negative. Of 30 HIV-infected patients with weekly stool cultures performed after discharge, 29 (97%) were negative at week 1. Of 50 HIV-negative patients with weekly stool cultures, 49 (98%) were negative at week 1. In countries with endemic HIV infection, clinicians should consider screening patients presenting with suspected cholera for HIV coinfection. © The American Society of Tropical Medicine and Hygiene.

  11. Mechanisms of pertussis toxin-induced barrier dysfunction in bovine pulmonary artery endothelial cell monolayers.

    Science.gov (United States)

    Patterson, C E; Stasek, J E; Schaphorst, K L; Davis, H W; Garcia, J G

    1995-06-01

    We have previously characterized several G proteins in endothelial cells (EC) as substrates for the ADP-ribosyltransferase activity of both pertussis (PT) and cholera toxin and described the modulation of key EC physiological responses, including gap formation and barrier function, by these toxins. In this study, we investigated the mechanisms involved in PT-mediated regulation of bovine pulmonary artery endothelial cells barrier function. PT caused a dose-dependent increase in albumin transfer, dependent upon action of the holotoxin, since neither the heat-inactivated PT, the isolated oligomer, nor the protomer induced EC permeability. PT-induced gap formation and barrier dysfunction were additive to either thrombin- or thrombin receptor-activating peptide-induced permeability, suggesting that thrombin and PT utilize distinct mechanisms. PT did not result in Ca2+ mobilization or alter either basal or thrombin-induced myosin light chain phosphorylation. However, PT stimulated protein kinase C (PKC) activation, and both PKC downregulation and PKC inhibition attenuated PT-induced permeability, indicating that PKC activity is involved in PT-induced barrier dysfunction. Like thrombin-induced permeability, the PT effect was blocked by prior increases in adenosine 3',5'-cyclic monophosphate. Thus PT-catalyzed ADP-ribosylation of a G protein (possibly other than Gi) may regulate cytoskeletal protein interactions, leading to EC barrier dysfunction.

  12. 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.

  13. Estudo da radiosensibilidade ao 60CO do Vibrio cholerae O1 incorporado em ostras Radiosensibility of Vibrio cholerae o1 incorporated in oysters, to 60CO

    Directory of Open Access Journals (Sweden)

    Ivany R de Moraes

    2000-02-01

    Full Text Available OBJETIVO: Avaliar a eficiência da radiação ionizante por 60CO na eliminação de Vibrio cholerae O1, El Tor Ogawa, não-toxigênico, incorporados laboratorialmente em ostras vivas da espécie Crassostrea brasiliana. MÉTODO: Foram selecionadas amostras de ostras provenientes de Canan��ia (litoral sul de São Paulo, Brasil, as quais foram contaminadas com Vibrio cholerae e irradiadas com doses de 0,5 kGy e 1,0 kGy. RESULTADOS: Foram observadas diminuições significativas do número inicial do microrganismo indicado: de 3,4.10(7 para 10³ e 10², respectivamente. Os valores de D10 correspondentes foram de 0,173 a 0,235. CONCLUSÃO: Adotando-se o fator 6 como nível de segurança, conclui-se que a dose de irradiação de 1,41 kGy é necessária para eliminar números elevados de células viáveis de V. cholerae em ostras. Os experimentos foram realizados com os controles respectivos.OBJECTIVE: Evaluate the effect of ionizing irradiation by 60Co on Vibrio cholerae O1, El-Tor, Ogawa, non-toxigenic, incorporated in live oysters Crassostrea brasiliana. METHODS: Samples of oysters were selected from Cananéia town in the South coast of S. Paulo state, Brazil, contaminated with Vibrio cholerae and irradiated with 60Co at 0.5 and 1.0 kGy dosages. RESULTS: Showed significant reductions of the initial number of V. cholerae , ranging from 3.4 x10(7 to 10³ and 10², respectively. The D10 values related with the respective doses of irradiation were 0.173 and 0.235. CONCLUSION: Considering a 6 value as safety factor, it is concluded that 1.41 kGy irradiation dosage is necessary to eliminate a high number of V. cholerae viable cells in oysters. Controls were used in the experiment.

  14. Respon Imun Anak Babi Pasca Vaksinasi Hog Cholera

    Directory of Open Access Journals (Sweden)

    I Made Jayanata

    2016-10-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui pengaruh antibodi maternal terhadap titer antibodi anak babi yang di vaksin hog cholera umur 7 hari. Penelitian menggunakan tujuh sampel babi dari induk yang divaksin secara teratur yang diberikan perlakuan vaksinasi pada umur 7 hari. Pengambilan sampel serum dilakukan pravaksinasi (7 hari, dan satu minggu serta dua minggu pasca vaksinasi. Untuk menentukan titer antibodi virus Hog cholera pada sampel anak babi dilakukan uji ELISA. Data yang diperoleh kemudian dianalisis mengunakan paired sampel T test antara titer antibodi hog cholera. Hasil paired sample T test menunjukkan terjadinya penurunan titer antibodi maternal yang nyata (p<0,05 pada pra vaksinasi ( umur 7 hari dengan satu minggu pasca vaksinasi dan sangat nyata (p<0,01 dengan hari dua minggu pasca vaksinasi. Dari hasil penelitian ini dapat disimpulkan bahwa antibodi maternal yang tinggi akan mengakibatkan penurunan pada titer antibodi pasca vaksinasi. Perlu dilakukan penelitian lebih lanjut untuk mengetahui waktu vaksinasi yang efektif

  15. Review of two decades of cholera diagnostics--how far have we really come?

    Directory of Open Access Journals (Sweden)

    Michal H Dick

    Full Text Available Cholera, an ancient scourge, continues to inflict high rates of mortality today. The rising incidence of epidemics in areas of poor sanitation and crowding highlight the need for better epidemic prevention and early response. Such interventions require the availability of rapid and accurate diagnostic techniques to trigger timely response and mitigate the scale of the outbreak. The current gold standard of bacterial culture is inadequate for rapid diagnosis, highlighting the overarching neglect of field diagnostic needs. This paper was written to support the World Health Organisation's Global Task Force on Cholera Control mandated Cholera and diarrhoeal disease laboratory Network (CholdiNet in devising a protocol for the validation of Rapid Diagnostic Tests (RDTs for Vibrio cholerae. The status of diagnostic tools for Vibrio cholerae is assessed, describing products that have been commercialised over the last two decades and discussing their peer-reviewed evaluation.Review of post-1990 peer-reviewed and grey literature on rapid diagnostic tests for Vibrio cholerae.Since 1990, twenty four diagnostic tests have been developed for the detection of Vibrio cholerae in human faecal samples. Fourteen of these have also been described in the literature, with rapid chromatographic-immuno assays (CIA featuring strongly. Polymerase chain reaction (PCR assays maintain the ability to detect the lowest amount of bacteria; however CIAs achieve both low detection thresholds and high sensitivity and specificity, making them possible candidates for use in field conditions. Field and laboratory studies were performed in a wide range of settings demonstrating variability in performance, however only a few of these studies were sufficiently stringent, highlighting five RDTs that showed promise in field conditions; COAT, IP cholera dipstick, SMART, IP dipstick and Medicos. In light of non-independent reporting, the authors would like to see these five products

  16. Review of two decades of cholera diagnostics--how far have we really come?

    Science.gov (United States)

    Dick, Michal H; Guillerm, Martine; Moussy, Francis; Chaignat, Claire-Lise

    2012-01-01

    Cholera, an ancient scourge, continues to inflict high rates of mortality today. The rising incidence of epidemics in areas of poor sanitation and crowding highlight the need for better epidemic prevention and early response. Such interventions require the availability of rapid and accurate diagnostic techniques to trigger timely response and mitigate the scale of the outbreak. The current gold standard of bacterial culture is inadequate for rapid diagnosis, highlighting the overarching neglect of field diagnostic needs. This paper was written to support the World Health Organisation's Global Task Force on Cholera Control mandated Cholera and diarrhoeal disease laboratory Network (CholdiNet) in devising a protocol for the validation of Rapid Diagnostic Tests (RDTs) for Vibrio cholerae. The status of diagnostic tools for Vibrio cholerae is assessed, describing products that have been commercialised over the last two decades and discussing their peer-reviewed evaluation. Review of post-1990 peer-reviewed and grey literature on rapid diagnostic tests for Vibrio cholerae. Since 1990, twenty four diagnostic tests have been developed for the detection of Vibrio cholerae in human faecal samples. Fourteen of these have also been described in the literature, with rapid chromatographic-immuno assays (CIA) featuring strongly. Polymerase chain reaction (PCR) assays maintain the ability to detect the lowest amount of bacteria; however CIAs achieve both low detection thresholds and high sensitivity and specificity, making them possible candidates for use in field conditions. Field and laboratory studies were performed in a wide range of settings demonstrating variability in performance, however only a few of these studies were sufficiently stringent, highlighting five RDTs that showed promise in field conditions; COAT, IP cholera dipstick, SMART, IP dipstick and Medicos. In light of non-independent reporting, the authors would like to see these five products undergoing

  17. Chitin-induced T6SS in Vibrio cholerae is dependent on ChiS activation.

    Science.gov (United States)

    Chourashi, Rhishita; Das, Suman; Dhar, Debarpan; Okamoto, Keinosuke; Mukhopadhyay, Asish K; Chatterjee, Nabendu Sekhar

    2018-05-01

    Vibrio cholerae regularly colonizes the chitinous exoskeleton of crustacean shells in the aquatic region. The type 6 secretion system (T6SS) in V. cholerae is an interbacterial killing device. This system is thought to provide a competitive advantage to V. cholerae in a polymicrobial community of the aquatic region under nutrient-poor conditions. V. cholerae chitin sensing is known to be initiated by the activation of a two-component sensor histidine kinase ChiS in the presence of GlcNAc2 (N,N'-diacetylchitobiose) residues generated by the action of chitinases on chitin. It is known that T6SS in V. cholerae is generally induced by chitin. However, the effect of ChiS activation on T6SS is unknown. Here, we found that ChiS inactivation resulted in impaired bacterial killing and reduced expression of T6SS genes. Active ChiS positively affected T6SS-mediated natural transformation in V. cholerae. ChiS depletion or inactivation also resulted in reduced colonization on insoluble chitin surfaces. Therefore, we have shown that V. cholerae colonization on chitinous surfaces activates ChiS, which promotes T6SS-dependent bacterial killing and horizontal gene transfer. We also highlight the importance of chitinases in T6SS upregulation.

  18. 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.

  19. Antimicrobial drugs for treating cholera.

    Science.gov (United States)

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

    2014-06-19

    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. To quantify the benefit of antimicrobial treatment for patients with cholera, and determine whether there are differences between classes of antimicrobials or dosing schedules. We searched the Cochrane Infectious Disease Group Specialized Register; the Cochrane Central Register of Controlled Trials (CENTRAL); PubMed; EMBASE; African Index Medicus; LILACS; Science Citation Index; metaRegister of Controlled Trials; WHO International Clinical Trials Registry Platform; conference proceedings; and reference lists to March 2014. Randomized and quasi-randomized controlled clinical trials in adults and children with cholera that compared: 1) any antimicrobial treatment with placebo or no treatment; 2) different antimicrobials head-to-head; or 3) different dosing schedules or different durations of treatment with the same antimicrobial. Two reviewers independently applied inclusion and exclusion criteria, and extracted data from included trials. Diarrhoea duration and stool volume were defined as primary outcomes. We calculated mean difference (MD) or ratio of means (ROM) for continuous outcomes, with 95% confidence intervals (CI), and pooled data using a random-effects meta-analysis. The quality of evidence was assessed using the GRADE approach. Thirty-nine trials were included in this review with 4623 participants. Antimicrobials versus placebo or no treatment Overall, antimicrobial therapy shortened the mean duration of diarrhoea by about a day and a half compared to placebo or no treatment (MD -36.77 hours, 95% CI -43.51 to -30.03, 19 trials, 1013 participants, moderate quality evidence). Antimicrobial therapy also

  20. GM1 erythroimmunoassay for detection and titration of Escherichia coli heat-labile enterotoxin.

    OpenAIRE

    Germani, Y; Bégaud, E; Guesdon, J L; Moreau, J P

    1986-01-01

    A GM1 ganglioside erythroimmunoassay for the detection of heat-labile Escherichia coli enterotoxin (LT) was developed for use in poorly equipped laboratories in developing countries. This assay is based on the immunological similarity between Vibrio cholerae toxin and LT and uses cholera toxin antiserum and sheep anti-rabbit immunoglobulin covalently coupled to sheep erythrocytes as conjugate. This assay has the following advantages over other currently available techniques: the reagents it u...