Ecological study of Vibrio cholerae in Vellore.
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Vellore is endemic for cholera due to Vibrio cholerae O1 and O139. In a previous study the prevalence of Vibrio cholerae in drinking water, lakes and sewage outfalls in a single 2-months period in Vellore, India was determined. In addition water samples from three sites were also tested for the presence of V. cholerae O1 and O139 by fluorescent antibody staining. This follow on study has examined how the environmental distribution of V. cholerae at the same sites alters over a 12-month period and the relationship to the clinical pattern of cholera in Vellore. Samples of water were collected from fixed sites at three water bodies each month between April 1997 and March 1998. Bacteria isolated from samples were identified by standard biochemical tests and isolated strains of V. cholerae tested for their ability to agglutinate O1 and O139 antisera. Samples were also tested for the presence of V. cholerae O1 and O139 by fluorescent antibody staining. The clinical isolation rate of V. cholerae in Vellore, maximum temperature and rainfall were also studied. The results demonstrate the presence in the environment of viable but non-cultivable (VNC) V. cholerae in 10 of 12 months of the study year as well as their viability. Their prevalence in the environment also correlated with the isolation of these pathogens from clinical samples over the same study period. (+info)
Two-year study of the protective efficacy of the oral whole cell plus recombinant B subunit cholera vaccine in Peru.
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The protective efficacy of an oral inactivated whole cell Vibrio cholerae plus recombinant B subunit cholera vaccine was determined against El Tor cholera among Peruvian children and adults (2-65 years old) in a randomized, double-blind manner. Study subjects received 2 doses of vaccine or placebo 2 weeks apart, followed by a booster dose 10 months later. Surveillance for cholera was performed actively, with 2 visits per week to each household, and passively, at a local hospital. Stool samples were collected during diarrhea episodes and were cultured for V. cholerae. A total of 17,799 persons received 2 doses of vaccine or placebo, and 14,997 of these persons received the booster dose. After 2 doses (first surveillance period), V. cholerae biotype O1 was isolated from 17 vaccinees and 16 placebo recipients, demonstrating vaccine efficacy (VE) of -4%. After 3 doses (second surveillance period), V. cholerae O1 was isolated from 13 vaccinees and 32 placebo recipients, demonstrating VE of 61% (95% confidence interval inverted question markCI, 28%-79%). In the second surveillance period, the VE for illness requiring hospitalization was 82% (95% CI, 27%-96%). VE was also higher for persons >15 years old (VE, 72%; 95% CI, 28%-89%). (+info)
Use of RNA arbitrarily primed-PCR fingerprinting to identify Vibrio cholerae genes differentially expressed in the host following infection.
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Evidence suggests that a repertoire of Vibrio cholerae genes are differentially expressed in vivo, and regulation of virulence factors in vivo may follow a different pathway. Our work was aimed at characterization of in vivo-grown bacteria and identification of genes that are differentially expressed following infection by RNA arbitrarily primed (RAP)-PCR fingerprinting. The ligated rabbit ileal loop model was used. The motility of in vivo-grown bacteria increased by 350% over that of in vitro-grown bacteria. Also, the in vivo-grown cells were more resistant to killing by human serum. By using the RAP-PCR strategy, five differentially expressed transcripts were identified. Two in vitro-induced transcripts encoded polypeptides for the leucine tRNA synthatase and the 50S ribosomal protein, and the three in vivo-induced transcripts encoded the SucA and MurE proteins and a polypeptide of unknown function. MurE is a protein involved in the peptidoglycan biosynthetic pathway. The lytic profiles of in vivo- and in vitro-grown cells suspended in distilled water were compared; the former was found to be slightly less sensitive to lysis. Ultrathin sections of both cells observed under the transmission electron microscope revealed that in contrast to the usual wavy discontinuous membrane structure of the in vitro-grown cells, in vivo-grown cells had a more rigid, clearly visible double-layered structure. The V. cholerae murE gene was cloned and sequenced. The sequence contained an open reading frame of 1,488 nucleotides with its own ribosome-binding site. A plasmid containing the murE gene of V. cholerae was transformed into V. cholerae 569B, and a transformed strain, 569BME, containing the plasmid was obtained. Ultrathin sections of 569BME viewed under a transmission electron microscope revealed a slightly more rigid cell wall than that of wild-type 569B. When V. cholerae 569B and 569BME cells were injected separately into ligated rabbit ileal loops, the transformed cells had a preference for growth in the ileal loops versus laboratory conditions. (+info)
Effect of sulfadoxine on transmission of Vibrio cholerae infection among family contacts of cholera patients in Calcutta.
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Sulfadoxine, a long-acting sulfonamide, and tetracycline were compared as regards their effectiveness in reducing transmission of cholera infection among the contacts of cholera patients in Calcutta. A total of 109 healthy family contacts of confirmed hospitalized cholera patients were treated with a single oral dose of sulfadoxine graded according to age. Another similar group of 101 contacts received 6 divided doses of oral tetracycline over a period of 3 days. All these contacts were bacteriologically examined for 15 days. Results showed that tetracycline was effective in significantly reducing the load of cholera infection from the 2nd to 6th day, while sulfadoxine was effective from the 3rd to the 6th day. The advantages and disadvantages of the two drugs as chemoprophylactic agents in cholera are discussed. (+info)
Doxycycline in the treatment of cholera.
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Doxycycline was compared with tetracycline in the treatment of cholera. Four types of treatment were compared: Group A was given 200 mg of doxycycline on admission and 100 mg on the second day; Group B was given 200 mg of doxycycline on admission only; Group C was given 300 mg of doxycycline on admission only; and Group D received 500 mg of tetracycline every 6 h for 48 h. Tetracycline showed a slight advantage in respect of duration of diarrhoea and vibrio excretion compared with doxycycline given as a single dose of 300 mg, but fluid intake and output were about the same in these two groups. The other two doxycycline treatment schedules did not compare well with tetracycline treatment. (+info)
Value of phage typing of Vibrio cholerae biotype eltor in West Africa.
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The epidemiological value of phage typing of El Tor vibrios in West Africa was assessed by testing 211 representative strains from all outbreaks of the cholera epidemic in Togo (1970-73) with the prophage typing method of Nicolle et al. and the lytic El Tor phages of Basu & Mukerjee. Prophage typing proved to be of limited epidemiological value in Togo since 203 of the 211 strains tested belonged to the same phage type-namely, type 2 of Nicolle et al., which corresponds to the Celebes type of Takeya. Six strains belonged to type 1, 1 strain to type 4, and 1 strain was untypable.Tested by the lytic El Tor phages, 175 strains were found to belong to type 4 of Basu & Mukerjee, 35 strains to type 1, and 1 strain was untypable. All but two strains of type 1 were isolated from 3 outbreaks in which this phage type was found almost exclusively. The results are in good agreement with epidemiological data collected during the epidemic. El Tor phage V reacted with a specificity of over 99% when tested against 601 strains of El Tor and NCV vibrios from Togo and may be recommended for the rapid identification of El Tor vibrios. (+info)
Microbiological safety of water.
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Significant advances in water treatment over the last century have resulted in massive improvements in the microbiological safety of public drinking water supplies in the UK and the developed countries. Incidences of illness due to poor treatment or post-treatment contamination are rare, but when they occur tend to attract considerable media attention. A well managed water treatment works and supply system can provide high quality drinking water wherever in the world it is located. As a rule, throughout the world, private supplies tend to be of a poorer quality than public supplies, but poorly managed public supplies have the potential to make a large number of people ill and continued effort is needed to maintain and improve drinking water quality world-wide. (+info)
Sunlight-induced propagation of the lysogenic phage encoding cholera toxin.
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In toxigenic Vibrio cholerae, the cholera enterotoxin (CT) is encoded by CTXPhi, a lysogenic bacteriophage. The propagation of this filamentous phage can result in the origination of new toxigenic strains. To understand the nature of possible environmental factors associated with the propagation of CTXPhi, we examined the effects of temperature, pH, salinity, and exposure to direct sunlight on the induction of the CTX prophage and studied the transmission of the phage to potential recipient strains. Exposure of cultures of CTXPhi lysogens to direct sunlight resulted in approximately 10,000-fold increases in phage titers. Variation in temperature, pH, or salinity of the culture did not have a substantial effect on the induction of the prophage, but these factors influenced the stability of CTXPhi particles. Exposure of mixed cultures of CTXPhi lysogens and potential recipient strains to sunlight significantly increased both the in vitro and in vivo (in rabbit ileal loops) transduction of the recipient strains by CTXPhi. Included in these transduction experiments were two environmental nontoxigenic (CTXPhi(-)) strains of V. cholerae O139. These two O139 strains were transduced at high efficiency by CTXPhi, and the phage genome integrated into the O139 host chromosome. The resulting CTXPhi lysogens produced biologically active CT both in vitro and in rabbit ileal loops. This finding suggests a possible mechanism explaining the origination of toxigenic V. cholerae O139 strains from nontoxigenic progenitors. This study indicates that sunlight is a significant inducer of the CTX prophage and suggests that sunlight-induced transmission of CTXPhi may constitute part of a natural mechanism for the origination of new toxigenic strains of V. cholerae. (+info)