Exploring the dynamics of toxigenic environmental Vibrio mimicus and its comparative analysis with Vibrio cholerae of the southern Gangetic delta.

Vibrio mimicus and Vibrio cholerae are closely related species. Environmental V.mimicus were comparatively analyzed with V.cholerae, for the presence of virulence genes, antibiotic susceptibility, resistance genes, in-vitro hemolysis, and biofilm formation. Phylogenetic analysis was performed depending on toxin-gene disposition and isolation area. One V.mimicus isolate harbored ctxA, tcp El-Tor, toxT and toxS, whereas several strains contained incomplete copies of virulence cassettes and associated toxin genes. V.cholerae isolates harbored ctx, tcp and toxT genes, with a higher preponderance of hlyA, rtxA and toxR genes. V.mimicus were highly sensitive to amino/carboxy-penicillins, furazolidone & gentamycin, with quinolone & tetracycline resistance genes. V.cholerae isolates were sensitive to penicillins and cephalosporins, with 29% of the strains bearing the sxt gene. Phylogenetically, the apomorphic strains of both species were unique to the inland sites. V.cholerae has embodied an enormous public health burden globally but our findings emphasize the role of V.mimicus as an emerging etiological agent with similar epidemic potential.

Seasonal influence, enteropathogenic microbial load and diarrhoeal enigma in the Gangetic Delta, India: Present scenario and health implications.

BACKGROUND: Environmental diarrheagenic enteropathogens, effect of surrounding attributes and disease dynamicity remains far from being conclusively explored. Population flux, poor sanitation and hygienic practice poses potential health threat in diarrhoea endemic tropical countries like India. We aim to identify environmental attributes, seasonality of water-borne enteropathogens and health risk assessment off the river Ganges. METHODS: A yearlong sampling data generated from three sites on either sides of the River was analysed and implications have been reported. Immediately after sample collection, physico chemical and bacterial indices were measured at the sampling site and laboratory respectively, followed by further statistical analysis of the findings. RESULTS: Annual variation of physico-chemical indices viz., temperature 18°C-36°C, pH 7.49-8.67, conductivity 215-468µS/cm and turbidity 25.6-593 NTU was recorded in the riverine water samples. High temperature and turbidity were recorded in the summer and monsoon at all sites. High bacterial dispersion has been positively correlated with turbidity and temperature variation (P<0.01; P<0.1) as we report TBC 103-105CFU/ml, TCC 103-104CFU/ml and CVC 4-212CFU/ml, with higher distribution in the monsoon and reverse in the winter. This suggests that the bacterial pool proliferates at higher temperature whereas turbidity enhances their survival providing the substratum for the bacterial pool. CVC could be positively correlated with conductivity which implies that ionic content of water augments the Vibrio load. Adaptive capability of Vibrios to sustain in very low saline riverine setting seems to be assisted by turbid water coupled with nutrient rich organic matter. CONCLUSION: Our present work establishes the interplay of seasonal variants on the dynamicity of enteropathogenic bacteria in flowing aquatic ecosystem. It also categorises the existing microbial threats in the Ganga River to help monitor the conventional as well as emerging diarrhoeal pathogens to reduce diarrheal recurrences.

Waterborne outbreaks in diarrhoea endemic foci of India: a longitudinal exploration and its implications.

Diarrhoea remains a global public health enigma raising deep concerns for the health planners since contaminated potable water often spoils the community health structure. We hereby report a 6-year odd continuing outbreak surveillance report based on potable water indices, during which 264 water samples were screened from different districts of West Bengal, India. Samples were analysed for the presence of different enteropathogenic bacterial species by conventional molecular tools and their sensitivity to antibiotics. 78.03% samples were positive for enteropathogenic bacterial organisms and 75% samples harbored Coliform. 45.45, 12.12, and 4.16% samples were positive for E.coli, V. cholerae, and V. mimicus, respectively. Diarrhoeagenic E.coli 7 EPEC, 10 ETEC, and 2 EIEC were isolated along with 2 V. cholerae O1 Ogawa (ctxA and tcpA ElTor positive), one each from tube well and pond. Interestingly, 4 V.cholerae non-O1/non-O139 also harbored hlyA gene. The detection of toxin genes among this bacterial pool of sampled water indicates the fallout of the potable water sources, thus enabling us to establish that it is none other than the contaminated potable water system which often wreaks havoc in the south Bengal diarrhoeal menace. The consequences are further complicated by the presence of drug-resistant pathogenic bacterial pool to fluoroquinolone, beta-lactams, and cephalosporins, in the accessible potable water, with threats of outbreaks exploding into an epidemic, given suitable environment, poor sanitation, and unhygienic practices. Therefore, we strongly recommend re-modelling of 'point-of-use water disinfection' measures and adequate personal hygiene for healthier community life.

Environmental Vibrio cholerae non O1/ non O139 from the Gangetic delta: a diarrhoeal disease purview.

Diarrhoea still remains an unsolved enigma in developing countries, a major concern for the health planners. We targeted the abundance and toxicity of Vibrio cholerae non-O1/non-O139 (NOVC) in Gangetic riverine-estuarine ecosystem. A total of 74 V. cholerae were isolated from 120 water samples (68 NOVC, 6 V. cholerae O1) from two sampling sites off river Ganges. V. cholerae showed distinct seasonality, with steady increase from summer to monsoon, steep ascent in post-monsoon and an abrupt decline in winter. Highest number of NOVC was isolated form Howrah, attributed to low salinity and high anthropogenic influence. Environmental NOVC harboured hlyA (94.0 %), rtxA (81.0 %) and toxR (28.0 %) genes. About 23.4 % of the hlyA harbouring NOVC showed haemolytic activity. Accessory toxin genes (tlcR, toxT, RJ and LJ and aldA), among 3-5 % of the NOVC carry significant health implications. Haemolytic activity and biofilm formation in NOVC, during unfavourable conditions, facilitates gene transfer and emphasises the role of environmental NOVC in diarrhoeal incidence in South Bengal, India.

Specificity of coliphages in evaluating marker efficacy: a new insight for water quality indicators.

Conventional procedures for qualitative assessment of coliphage are time consuming multiple step approach for achieving results. A modified and rapid technique has been introduced for determination of coliphage contamination among potable water sources during water borne outbreaks. During December 2013, 40 water samples from different potable water sources, were received for water quality analyses, from a jaundice affected Municipality of West Bengal, India. Altogether, 30% water samples were contaminated with coliform (1-20 cfu/ml) and 5% with E. coli (2-5 cfu/ml). Among post-outbreak samples, preponderance of coliform has decreased (1-4 cfu/ml) with total absence of E. coli. While standard technique has detected 55% outbreak samples with coliphage contamination, modified technique revealed that 80%, double than that of bacteriological identification rate, were contaminated with coliphages (4-20 pfu/10 ml). However, post-outbreak samples were detected with 1-5 pfu/10 ml coliphages among 20% samples. Coliphage detection rate through modified technique was nearly double (50%) than that of standard technique (27.5%). In few samples (with coliform load of 10-100 cfu/ml), while modified technique could detect coliphages among six samples (10-20 pfu/10 ml), standard protocol failed to detect coliphage in any of them. An easy, rapid and accurate modified technique has thereby been implemented for coliphage assessment from water samples. Coliform free water does not always signify pathogen free potable water and it is demonstrated that coliphage is a more reliable 'biomarker' to ascertain contamination level in potable water.