Survival potential of non-culturable vibrio cholerae 01 by laboratory

Abstract

Objective: Assess the survival potential of V, cholerae 01 using conventional cultural, fluorescent antibody and recently developed polymerase chain reaction techniques. In Bangladesh, cholera is endemic in certain areas and flares into seasonal epidemics. V. cholerae 01 is one of the causative agents of cholera. The survival potential of V. cholerae 01 was carried out in various environmental samples by using conventional techniques. Methods: The strain V. cholerae Ol biotype El Tor serotype Ogawa was used in this study carried out at the ICDDR,B's laboratories in Dhaka, Bangladesh. A measured inoculum of about 1(P V. cholerae 01 per ml was added to 100 mi autoclaved pond water in a 500-ml conical flask, mixed and stored at room temperature. Culturable cells were counted on gelatin agar (GA) and taurocholate tellurite gelatin agar at various time intervals until the bacteria were no longer culturable. Results: The non-culturable V. cholerae 01 was detected by fluorescent antibody and PCR techniques. The culturable V. cholerae Ol was isolated up to 44 days from the pond water microcosms. The non-culturable V. cholerae 01 was detected up to 7 weeks by FA and PCR techniques after they lost their culturability. Conclusions: The non-culturable stage reported here for V. cholerae 01 is significant for understanding the epidemiology of cholera because the non-culturable state of V. cholerae Ol may pose health problems. Volunteer studies have shown that non-culturable V. cholerae Ol became culturable in volunteers intestine. This study demonstrated the survival of non-culturable V. cholerae 01 in surface water which may be important from the view point of public health