Vibrio parahaemolyticus has two flagellar systems, the polar flagellum and lateral flagella, which are both intricately regulated by a multitude of factors.
CalR, a LysR-type transcriptional regulator, is sensitive to calcium (Ca) and plays a crucial role in regulating the virulence and swarming motility of V.
parahaemolyticus. In this study, we have demonstrated that the deletion of calR significantly enhances the swimming motility of V. parahaemolyticus under low Ca conditions but not under high Ca conditions or in the absence of Ca. CalR binds to the regulatory DNA regions of flgM, flgA, and flgB, which are located within the polar flagellar gene loci, with the purpose of repressing their transcription. Additionally, it exerts an indirect negative control over the transcription of flgK. The overexpression of CalR in Escherichia coli resulted in a reduction in the expression levels of flgM, flgA, and flgB, while having no impact on the expression of flgK. In summary, this research demonstrates that the negative regulation of V. parahaemolyticus swimming motility by CalR under low Ca conditions is achieved through its regulation on the transcription of polar flagellar genes.
Community-based microbial source tracking (MST) can be
used to determine fecal contamination from multiple sources
in the aquatic environment. However, there is little scientific
information on its application potential in water environmental
management. Here, we compared SourceTracker and
Fast Expectation-maximization Microbial Source Tracking
(FEAST) performances on environmental water bodies exposed
to low fecal pollution and evaluated treatment effects
of fecal pollution in the watershed utilizing community-based
MST. Our results showed that FEAST overall outperformed
SourceTracker in sensitivity and stability, and was able to discern
multi-source fecal contamination (mainly chicken feces)
in ambient water bodies exposed to low fecal inputs. Consistent
with our previous PCR/qPCR-based MST assays, FEAST
analysis indicates that fecal pollution has been significantly
mitigated through comprehensive environmental treatment
by the local government. This study suggests that FEAST can
be a powerful tool for accurately evaluating the contribution
of multi-source fecal contamination in environmental water,
facilitating environmental management.
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In order to adapt to different environments, Vibrio parahaemolyticus
employed a complicated quorum sensing system to
orchestrate gene expression and diverse colony morphology
patterns. In this study, the function of the putative quorum
sensing signal synthase gene cqsA (VPA0711 in V. parahaemolyticus
strain RIMD2210633 genome) was investigated.
The cloning and expression of V. parahaemolyticus cqsA in
Escherichia coli system induced the production of a new quorum
sensing signal that was found in its culture supernatant.
The signal was purified by high performance liquid chromatography methods and determined to be 3-hydroxyundecan-
4-one by indirect and direct mass spectra assays. The deletion
of cqsA in RIMD2210633 changed V. parahaemolyticus
colony morphology from the classical ‘fried-egg’ shape (thick
and opaque in the center, while thin and translucent in the
edge) of the wild-type colony to a ‘pancake’ shape (no significant
difference between the centre and the edge) of the cqsAdeleted
colony. This morphological change could be restored
by complementary experiment with cqsA gene or the signal
extract. In addition, the expression of opaR, a well-known
quorum sensing regulatory gene, could be up-regulated by
cqsA deletion. Our results suggested that V. parahaemolyticus
used cqsA to produce 3-hydroxyundecan-4-one signal
and thereby regulated colony morphology and other quorum
sensing-associated behaviors.
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Vibrio parahaemolyticus is a halophilic bacterium endemic
to coastal areas, and its pathogenicity has caused widespread
seafood poisoning. In our previous research, the protein expression
of V. parahaemolyticus in Fe3+ medium was determined
using isobaric tags for relative and absolute quantitation
(iTRAQ). Here, nuclear magnetic resonance (NMR) was
used to detect changes in the V. parahaemolyticus metabolome.
NMR spectra were obtained using methanol-water extracts
of intracellular metabolites from V. parahaemolyticus under
various culture conditions, and 62 metabolites were identified,
including serine, arginine, alanine, ornithine, tryptophan,
glutamine, malate, NAD+, NADP+, oxypurinol, xanthosine,
dCTP, uracil, thymine, hypoxanthine, and betaine. Among
these, 21 metabolites were up-regulated after the stimulation
of the cells by ferric iron, and 9 metabolites were down-regulated.
These metabolites are involved in amino acid and protein
synthesis, energy metabolism, DNA and RNA synthesis
and osmolality. Based on these results, we conclude that Fe3+
influences the metabolite profiles of V. parahaemolyticus.
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Vibrio parahaemolyticus, a Gram-negative marine bacterial pathogen, is emerging as a major cause of food-borne illnesses worldwide due to the consumption of raw seafood leading to diseases including gastroenteritis, wound infection, and septicemia. The bacteria utilize toxins and type III secretion system (T3SS) to trigger virulence. T3SS is a multi-subunit needle-like apparatus used to deliver bacterial proteins, termed effectors, into the host cytoplasm which then target various eukaryotic signaling pathways. V. parahaemolyticus carries two T3SSs in each of its two chromosomes, named T3SS1 and T3SS2, both of which play crucial yet distinct roles during infection: T3SS1 causes cytotoxicity whereas T3SS2 is mainly associated with enterotoxicity. Each T3SS secretes a unique set of effectors that contribute to virulence by acting on different host targets and serving different functions. Emerging studies on T3SS2 of V. parahaemolyticus, reveal its regulation, translocation, discovery, characterization of its effectors, and development of animal models to understand the enterotoxicity. This review on recent findings for T3SS2 of V. parahaemolyticus highlights a novel mechanism of invasion that appears to be conserved by other marine bacteria.
Ingestion of shellfish-associated Vibrio parahaemolyticus is the primary cause of potentially severe gastroenteritis in many countries. However, only Kanagawa phenomenon (hemolysin) positive (KP^+) strains of V. parahaemolyticus are isolated from patients, whereas >99% of strains isolated from the environment do not produce this hemolysin (i.e. are KP^-). The reasons for these differences are not known. Following a temperature downshift, Vibrio parahaemolyticus enters the viable but nonculturable (VBNC) state wherein cells maintain viability but cannot be cultured on routine microbiological media. We speculated that KP^+ and KP^- strains may respond differently to the temperature and salinity conditions of seawater by entering into this state which might account for the low numbers of culturable KP^+ strains isolated from estuarine waters. The response of eleven KP^+ and KP^- strains of V. parahaemolyticus following exposure to a nutrient and temperature downshift in different salinities, similar to conditions encountered in their environment, was examined. The strains included those from which the KP^+ genes had been selectively removed or added. Our results indicated that the ability to produce hemolysin did not affect entrance into the VBNC state. Further, VBNC cells of both biotypes could be restored to the culturable state following an overnight temperature upshift.
Vibrio parahaemolyticus is a halophilic bacterium associated with seafood gastroenteritis. An unusual strain of Kanagawa-positive urease producing Vibrio parahaemolyticus O1 : K1 was isolated from the environment and identified. A polymerase chain reaction assay revealed that this strain harbored both the tdh and trh genes. The urease from this strain was studied. Maximum urease production was induced in LB medium containing 0.2% urea, 0.5% glucose, 2% NaCl and pH 5.5 with 6 h of cultivation at 37 C under aeration. Purification of urease was achieved by the process of whole cell lysate, 65% ammonium sulphate precipitation, DEAE-cellulose ion exchange column chromatography, Sepharose CL-6B gel filtration and oxirane activated Sepharose 6B-urea affinity chromatography with 203 fold purification and 2.2% yield. Analysis of the purified enzyme by SDS-PAGE demonstrated the presence of the subunits with a molecular weight of 85 kDa, 59 kDa, 41 kDa and the molecular weight for the native enzyme by nondenaturing PAGE and gel filtration chromatography was 255 kDa. The purified urease was stable at pH 7.5 and the optimal pH in HEPES buffer was 8.0. The enzyme was stable at 60 C for 2 h with a residual activity of 32%. The addition of 10 uM of NiCl_2 maintained stability for 30 min. The Km value of the purified enzyme was 35.6 mM in urea substrate. The TD_50 (median toxic dose) of the purified urease was 2.5 ug/ml on human leukemia cells.