Anwar Huq

Viable but nonculturable Vibrio cholerae O1 in biofilms in the aquatic environment and their role in cholera transmission

Vibrio cholerae persists in aquatic environments predominantly in a nonculturable state. In this study coccoid, nonculturable V. cholerae O1 in biofilms maintained for 495 days in Mathbaria, Bangladesh, pond water became culturable upon animal passage. Culturability, biofilm formation, and the wbe, ctxA, and rstR2 genes were monitored by culture, direct fluorescent antibody (DFA), and multiplex PCR. DFA counts were not possible after formation of biofilm. Furthermore, wbe, but not ctxA, were amplifiable, even after incubation for 54 and 68 days at room temperature (≈25°C) and 4°C, respectively, when no growth was detectable. Slower biofilm formation and extended culturability were observed for cultures incubated at 4°C, compared with ≈25°C, suggesting biofilm production to be temperature dependent and linked to loss of culturability. Small colonies appearing after incubation in microcosms for 54 and 68 days at 25°C and 4°C, respectively, were wbe positive and ctxA and rstR2 negative, indicating loss of bacteriophage CTXΦ. The coccoid V. cholerae O1 observed as free cells in microcosms incubated for 495 days could not be cultured, but biofilms in the same microcosms yielded culturable cells. It is concluded that biofilms can act as a reservoir for V. cholerae O1 between epidemics because of its long-term viability in biofilms. In contrast to biofilms produced in Mathbaria pond water, V. cholerae O1 in biofilms present in cholera stools and incubated under identical conditions as the Mathbaria pond water biofilms could not be cultured after 2 months, indicating that those V. cholerae cells freshly discharged into the environment are significantly less robust than cells adapted to environmental conditions.

Marine ecosystems and cholera

Historically, most of the major epidemics or outbreaks of cholera around the world have originated in coastal regions. The most dramatic of recent outbreaks of cholera occurred in India and Bangladesh in 1991, followed by an outbreak of cholera after almost a century without cholera in South America in 1991. Both of these recent epidemics were reported first in the coastal regions of India and Peru, respectively. Cholera epidemics are seasonal, occurring during the spring and fall months. Outbreaks of cholera in noncholera epidemic areas have been ascribed to travel and shipping activities, but there is compelling evidence that V. cholerae always is present in the aquatic environment and proliferates under nonepidemic conditions while attached to, or associated with, eucaryotic organisms. It is hypothesized that climate directly influences the incidence and geographic distribution of the cholera bacterium.

Seasonal Cholera from Multiple Small Outbreaks, Rural Bangladesh

Clinical and environmental Vibrio cholerae organisms collected from February 2004 through April 2005 were systematically isolated from 2 rural Bangladeshi locales. Their genetic relatedness was evaluated at 5 loci that contained a variable number of tandem repeats (VNTR). The observed minimal overlap in VNTR patterns between the 2 communities was consistent with sequential, small outbreaks from local sources.

Is protection against shigellosis induced by natural infection with Plesiomonas shigelloides

Shigellosis due to Shigella sonnei is rare among people growing up and living in developing countries; however, infections due to S sonnei becomes more common than those due to S flexneri as societies develop economically. The relation between risk of S sonnei infection and economic development may be explained by the exposure of developing-country populations to Plesiomonas shigelloides. P shigelloides is often found in surface water, and one serotype (serotype 17) possesses a cell-wall lipopolysaccharide identical to that of S sonnei. Thus, exposure to P shigelloides by drinking contaminated water may immunise populations to S sonnei. As economic development occurs, water quality improves and populations become susceptible to S sonnei. Although drinking water has many advantages, immunisation against S sonnei may be one benefit of traditional water sources.

The influence of environmental conditions on the production of pigment bySerratia marcescens

Serratia marcescens biovar A2/A6, isolated from an Indonesian freshwater source, was identified based on extensive morphological, biochemical and genetic characterization. Formation of pigment was found to be strongly influenced by environmental conditions. Placket-Burman design was used to analyze the effect of carbon and nitrogen sources. Based on results of physiological and biochemical studies, the optimum conditions for growth and pigment formation were incubation 30°C in a neutral to slightly alkaline medium containing lactic acid and beef extract.

Occurrence and Expression of Luminescence in Vibrio cholerae

ABSTRACT Several species of the genus Vibrio, including Vibrio cholerae, are bioluminescent or contain bioluminescent strains. Previous studies have reported that only 10% of V. cholerae strains are luminescent. Analysis of 224 isolates of non-O1/non-O139 V. cholerae collected from Chesapeake Bay, MD, revealed that 52% (116/224) were luminescent when an improved assay method was employed and 58% (130/224) of isolates harbored the luxA gene. In contrast, 334 non-O1/non-O139 V. cholerae strains isolated from two rural provinces in Bangladesh yielded only 21 (6.3%) luminescent and 35 (10.5%) luxA+ isolates. An additional 270 clinical and environmental isolates of V. cholerae serogroups O1 and O139 were tested, and none were luminescent or harbored luxA. These results indicate that bioluminescence may be a trait specific for non-O1/non-O139 V. cholerae strains that frequently occur in certain environments. Luminescence expression patterns of V. cholerae were also investigated, and isolates could be grouped based on expression level. Several strains with defective expression of the lux operon, including natural K variants, were identified.

Rapid detection of Vibrio cholerae 01 in West Africa.

Most cholera epidemics begin in coastal areas lending credence to the hypothesis that the marine environment serves as a reservoir for the vibrio. In many cases viable but non-culturable Vibrio cholerae O1 may survive for some time in marine waters and then end dormancy with an unknown trigger resulting in an outbreak. This study evaluated the potential of two monitoring systems--Sensitive Membrane Antigen Rapid Test (SMART) and Cholera Direct Fluorescent Antibody (DFA)--to serve as an early warning system for outbreaks. Samples were collected in the summer of 1996 from a series of sites in the Ebrie lagoon in Abidjan Ivory Coast. The only sample positive for V cholerae O1 by the SMART test was the small mesh-sized particulate sample taken from Koumassi. The DFA gave a positive result for the Koumassi sample as well as those of small mesh-sized particulates from Yopougon. Koumassi and Yopougon are the two areas in Abidjan where the most cholera cases occur. The remaining low-mesh and all large mesh-sized particulates were negative for V cholerae O1. Further research on where in the food chain the vibrios are most concentrated their autochthonous presence in the lagoon and how the infection is spread to humans is necessary to facilitate development of a public health strategy for the control of cholera in the area.