Vibrio parahaemolyticus is the leading cause of seafood borne bacterial gastroenteritis in the world, often associated with the consumption of raw or undercooked seafood. However, not all strains of V. parahaemolyticus are pathogenic. The thermostable direct hemolysin (TDH) or TDH-related hemolysin (TRH) encoded by tdh and trh genes, respectively, are considered major virulence factors in V. parahaemolyticus. However, about 10% of clinical strains do not contain tdh and/or trh. Environmental isolates of V. parahaemolyticus lacking tdh and/or trh are also highly cytotoxic to human gastrointestinal cells. Even in the absence of these hemolysins, V. parahaemolyticus remains pathogenic indicating other virulence factors exist. This mini review aims at discussing the possible roles of tdh and trh genes in clinical and environmental isolates of V. parahaemolyticus.
Vibrio abundance generally displays seasonal patterns. In temperate coastal areas, temperature and salinity influence Vibrio growth, whereas in tropical areas this pattern is not obvious. The present study assessed the dynamics of Vibrio in the Arabian Sea, 1-2 km off Mangalore on the south-west coast of India, during temporally separated periods. The two sampling periods were signified by oligotrophic conditions, and stable temperatures and salinity. Vibrio abundance was estimated by culture-independent techniques in relation to phytoplankton community composition and environmental variables. The results showed that the Vibrio density during December 2007 was 10- to 100-fold higher compared with the February-March 2008 period. High Vibrio abundance in December coincided with a diatom-dominated phytoplankton assemblage. A partial least squares (PLS) regression model indicated that diatom biomass was the primary predictor variable. Low nutrient levels suggested high water column turnover rate, which bacteria compensated for by using organic molecules leaking from phytoplankton. The abundance of potential Vibrio predators was low during both sampling periods; therefore it is suggested that resource supply from primary producers is more important than top-down control by predators.
Aims: The main aims of this study were to clone and express an outer membrane protein (OMP), OmpW, of Aeromonas hydrophila and to study its distribution in Aeromonas spp.
Methods and Results: The gene encoding OmpW in A. hydrophila has been cloned and expressed in Escherichia coli. Primers were designed for amplification of full‐length ompW gene and used for identification of this gene in different Aeromonas spp. Of the 42 Aeromonas strains tested, all the isolates were positive by polymerase chain reaction (PCR) except one strain of Aeromonas veronii biovar veronii (VTE338). None of the other gram‐negative bacteria were positive by PCR with primers specific to ompW gene of A. hydrophila. Polyclonal antibodies were raised in rabbit against the purified recombinant protein and the reaction of these antibodies was confirmed by western blotting using the purified recombinant protein and 42 Aeromonas cultures grown at various salt concentrations.
Conclusions: The ompW‐based PCR method developed in this study was found to be 100% specific and 97% sensitive. Expression of OmpW protein of Aeromonas was found to be salt‐dependant. Recombinant OmpW protein was found to be highly immunogenic in fish.
Significance and Impact of the Study: To our knowledge, this is the first report on cloning and expression of OmpW protein of A. hydrophila. Full‐length ompW gene amplification by PCR can be used for the detection of Aeromonas. Recombinant OmpW protein can be useful for vaccination of fish against Aeromonas spp.
We report the results from a mesocosm study investigating the interrelationship between microalgae and vibrios. The mesocosms were inoculated with plankton, plankton + sediment, or sediment. We followed the diatom bloom and increases in the abundance of Vibrio spp. and V. parahaemolyticus in conjunction with several environmental variables in all mesocosms and at a reference site. The dominating diatom genera were also identified. Temperature, salinity, and pH were nearly invariant in the mesocosms and did not contribute to the results. The principal environmental variables that correlated to vibrio abundance were total bacterial plate counts, phosphorus and ammonia (positive relationship), and oxygen and silica (negative). Nitrate, total bacterial counts and chlorophyll a (chl a) did not correlate with vibrio growth. The highest diatom abundances were followed by increases in vibrios in all mesocosms. This was also observed in field sampling. Together, these results suggest that diatom blooms could support Vibrio spp. growth. V. parahaemolyticus was initially favoured by sediment. The contribution of V. parahaemolyticus to the total bacterial population was low, on average 0.5%, but constituted a rather high proportion of the vibrio population in the mesocosm systems, i.e. on average 18%. Some of the identified diatom genera, e.g. Chaetoceros and Skeletonema, were negatively correlated to vibrios, while Coscinodiscus was positively correlated. The results indicate that phytoplankton blooms, when recorded as high levels of chl a, should be used with caution as predictors for future vibrio epidemics, since the origin of the chl a might have a significant effect on vibrio abundance. KEY WORDS: Mesocosm · Phytoplankton blooms · Vibrio spp.
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OPEN PEN
Aims: Evaluation of protein profiling for typing Vibrio parahaemolyticus using 71 strains isolated from different seafood and comparison with other molecular typing techniques such as random amplified polymorphic DNA analysis (RAPD) and enterobacterial repetitive intergenic consensus sequence (ERIC)‐PCR.
Methods and Results: Three molecular typing methods were used for the typing of 71 V. parahaemolyticus isolates from seafood. RAPD had a discriminatory index (DI) of 0·95, while ERIC‐PCR showed a DI of 0·94. Though protein profiling had less discriminatory power, use of this method can be helpful in identifying new proteins which might have a role in establishment in the host or virulence of the organism.
Conclusions: The use of protein profiling in combination with other established typing methods such as RAPD and ERIC‐PCR generates useful information in the case of V. parahaemolyticus associated with seafood.
Significance and Impact of the Study: The study demonstrates the usefulness of nucleic acid and protein‐based studies in understanding the relationship between various isolates from seafood.
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