Laboratory methods in cholera: Isolation of vibrio cholerae (el tor and classical) on TCBS medium in minimally equipped laboratories

Gangarosa, Eugene J.; DeWitt, Wallis E.; Huq, I.; Zarifi, Abdolvahed

doi:10.1016/0035-9203(68)90121-1

Cited by 25 publications

(7 citation statements)

References 5 publications

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“…The results of this investigation allow us to conclude with confidence that 6-h APW enrichment offers no significant ad-vantage over 24-h APW enrichment for the isolation of V. cholerae O1 when the two methods are compared either as single procedures or as combined with direct plating of stool swabs on TCBS media. Also, in contrast to the opinion that the value of enrichment broth cultures may be limited to convalescing and/or antibiotic-treated patients who excrete V. cholerae in reduced numbers (5,6,12), we found that APW enrichment (for either 6 h or 24 h) significantly improves the recovery of V. cholerae from the acute cholera patient. Our results here were in agreement with those reported previously (9) which evaluated the use of APW enrichment for isolating V. cholerae and demonstrated that the combination of enrichment and direct TCBS plating was superior to direct plating alone.…”

Section: Discussioncontrasting

confidence: 99%

Comparison of direct-plating and enrichment methods for isolation of Vibrio cholerae from diarrhea patients

et al. 1997

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A direct-plating method on thiosulfate citrate bile salts sucrose agar (DIR-TCBS) in conjunction with enrichment in alkaline peptone water (APW) incubated for both 6 h and 24 h followed by subculture onto TCBS (APW6h-TCBS and APW24h-TCBS, respectively) was performed on 16,034 rectal swab samples for isolating Vibrio cholerae. A total of 2,932 (18.3%) rectal swab samples were positive for V. cholerae O1 biotype El Tor, with the Ogawa serotype constituting 99.2% of the isolates. There were no significant differences in V. cholerae O1 isolation rates between the three culture systems nor between the combinations of any two systems. However, direct plating plus enrichment demonstrated a significantly higher V. cholerae O1 isolation rate than DIR-TCBS alone (P < 0.02). Conversely, enrichment procedure, alone or in combination with DIR-TCBS, yielded significantly more (P < 0.0001) V. cholerae non-O1 isolates than DIR-TCBS alone. The length of incubation time of the enrichment broth, 6 h, offers no significant advantages over 24 h for the isolation of V. cholerae O1 and non-O1. A 24-h enrichment broth incubation period has the practical advantage of being easy to integrate into a normal laboratory workday, whereas 6-h broth enrichment, although more commonly recommended, requires that arrangements be made for after-hours subculture.

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Section: Discussioncontrasting

confidence: 99%

Comparison of direct-plating and enrichment methods for isolation of Vibrio cholerae from diarrhea patients

et al. 1997

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“…In all 3 of these treatment settings, all patients presenting with diarrhea were clinically evaluated and treated. For those residing in the ICDDR,B study area, fecal samples were collected and evaluated for V. cholerae, by serogroup, biotype, and serotype, using conventional microbiological methods [14,15]. All patient services in these facilities were free of charge, and there was a free boat service for patients who needed to be transported for care on an emergency basis.…”

Section: Study Areamentioning

confidence: 99%

Natural Cholera Infection-Derived Immunity in an Endemic Setting

Ali¹,

Emch²,

Park³

et al. 2011

Journal of Infectious Diseases

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Background. Live oral cholera vaccines may protect against cholera in a manner similar to natural cholera infections. However, information on which to base these vaccines is limited. Methods. The study was conducted in a cholera-endemic population in Bangladesh. Patients with cholera (index patients) detected between 1991 and 2000 were age-matched to 4 cholera-free controls and then followed up during the subsequent 3 years. Results. El Tor cholera was associated with a 65% (95% confidence interval [CI], 37%-81%; P , .001) lower risk of a subsequent El Tor episode. Reduction of the risk of subsequent El Tor cholera was similar for children ,5 years and for older persons and was sustained during all 3 years of follow-up. Having El Tor Inaba cholera was associated with lower risks of both El Tor Inaba and El Tor Ogawa cholera, but having El Tor Ogawa cholera was associated only with a reduced risk of El Tor Ogawa cholera. O139 cholera was associated with a 63% (95% CI, 261% to 92%; P 5 .18) lower risk of subsequent O139 cholera, but there was no evidence of cross-protection between the O1 and O139 serogroups. Conclusions. Live oral cholera vaccines designed to protect against the O1 and O139 serogroups should contain at least the Inaba serotype and strains of both serogroups.

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“…Excellent correlation was observed in test results for both TTGA and modified TTGA media. Results for the 78 vibrios examined on modified TTGA medium are presented in 11 (100) 1(100) 0 (0) 0 (0) 1 (100) 0 (0) 1 (100) 1 (100) V. fluvialis (5) 5 (100) 5 (100) 4 (80) 0 (0) 1 (20) 4 (80) 5 (100) 5 (100) V. harveyii (1) 1 (100) 0 (0) 0 (0) 1 (100) 0 (0) 0 (0) 1 (100) 1(100) V. metschnikoviid (1) 0 (0) 1 (100)…”

Section: Resultsmentioning

confidence: 99%

“…Taurocholatetellurite-gelatin agar (TTGA) medium, devised by Monsur (9) and recommended by the World Health Organization (15), has several distinct advantages over TCBS agar. It is relatively inexpensive to prepare and there are no problems in follow-up serological tests (4,10). Furthermore, oxidase activity can be determined directly from the plate (10).…”

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confidence: 99%

Modified taurocholate-tellurite-gelatin agar for improved differentiation of Vibrio species

O’Brien¹,

Colwell²

1985

J Clin Microbiol

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A total of 78 strains, representing 21 Vibrio species, were examined by using taurocholate-tellurite-gelatin agar (TTGA) medium and modified TTGA medium containing 4-methylumbelliferyl-I8-D-galactoside (150 ,ig/ml). Modified TTGA medium allowed for simple and direct detection of P-D-galactosidase (P-gal) activity. This feature, in conjunction with other differential characteristics of TTGA medium, gave improved differentiation of the vibrios tested. The modified TTGA medium allowed for easy differentiation of Vibrio cholerae (jP-gal') from Vibrioparahaemolyticus (P-gali). The 4-methylumbelliferyl-4-D-galactoside substrate is inexpensive and very stable. Incorporation into the agar did not affect the performance of TTGA as a * Corresponding author.

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Laboratory methods in cholera: Isolation of vibrio cholerae (el tor and classical) on TCBS medium in minimally equipped laboratories

Cited by 25 publications

References 5 publications

Comparison of direct-plating and enrichment methods for isolation of Vibrio cholerae from diarrhea patients

Comparison of direct-plating and enrichment methods for isolation of Vibrio cholerae from diarrhea patients

Natural Cholera Infection-Derived Immunity in an Endemic Setting

Modified taurocholate-tellurite-gelatin agar for improved differentiation of Vibrio species

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