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At least 19 recordsLinked to original sources
PubMed2005

Vibrio parahaemolyticus, Vibrio vulnificus and microorganisms of fecal origin in mussels (Mytilus galloprovincialis) sold in the Puglia region (Italy).

Mytilus galloprovincialis is one of the most commonly consumed of all bivalve molluscs. The consumption of raw bivalve molluscs has caused outbreaks of food poisoning due to Vibrio parahaemolyticus and Vibrio vulnificus. This paper reports the results of a survey on the presence of V. parahaemolyticus, V. vulnificus fecal coliform bacteria, Escherichia coli and Salmonella spp. in 600 M. galloprovincialis samples collected from retail outlets in the Puglia region. V. parahaemolyticus and V. vulnificus were found in 47 (7.83%) and 17 (2.83%) of the samples, respectively. One sample (0.16%) was contaminated with Salmonella spp. but no relationship was observed between vibrios and fecal coliforms and E. coli. There were no significant differences among vibrios present in bivalve molluscs during the 3-year survey.

Animals↗
PubMed1990

Genetic transformation of Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio cholerae non O-1 with plasmid DNA by electroporation.

An electroporation procedure for the plasmid-mediated transformation of the genus Vibrio was performed, as part of an effort to develop recombinant DNA techniques for genetic manipulation of the genus Vibrio. Vibrio parahaemolyticus, V. alginolyticus, and V. cholerae non O-1 (9 different strains) were transformed with 3 vector plasmids (pACYC184, pHSG398, and pBR325). The efficiency of transformation was highly dependent on three parameters: the concentration of plasmid DNA; the strength of the electric field; and the combination of plasmid DNA and recipient strain. The drug-resistance genes on the vector plasmid were expressed in the Vibrio strains.

DNA, Bacterial↗
PubMed2001

Identification and characterization of the sodA genes encoding manganese superoxide dismutases in Vibrio parahaemolyticus, Vibrio mimicus, and Vibrio vulnificus.

Sequencing of Fur titration assay-positive clones obtained from genomic DNA libraries of Vibrio parahaemolyticus, V. mimicus and V. vulnificus revealed open reading frames encoding proteins of 202, 205 and 202 amino acid residues, respectively. Each open reading frame was preceded by a predicted Fur box which overlaps a likely promoter with similarity to the -10 and -35 consensus sequence of Escherichia coli. The deduced amino acid sequences shared considerable homology with bacterial Mn-containing superoxide dismutases (MnSODs). Consistent with this, these Vibrio strains produced proteins with SOD activity resistant to inhibition by H2O2 and KCN only when grown under iron-limiting conditions. Primer extension analysis of the total RNA from these vibrios revealed iron-repressible expression of the genes. Furthermore, when grown under iron-limiting conditions, E. coli carrying a plasmid with each cloned gene overexpressed protein with the same electrophoretic mobility and insensitivity of SOD activity to H2O2 and KCN. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by N-terminal amino acid sequencing revealed that proteins (MnSODs) having N-terminal amino acid sequences consistent with those deduced from the corresponding genes were present in cell lysates of the vibrios grown under these iron-limited conditions. These results demonstrate that the genes cloned in this study are sodA homologs encoding MnSODs, whose expression is regulated by the iron status of the growth medium. PCR using a primer set based on the V. parahaemolyticus sodA sequence revealed the presence of homologous genes in certain other Vibrio species.

Amino Acid Sequence↗
PubMed1986

Report of a wound infection caused by Vibrio parahaemolyticus and Vibrio vulnificus.

The present case describes a foot wound caused by a clam shell from which both Vibrio parahaemolyticus and Vibrio vulnificus were recovered. Although extraintestinal infections associated with Vibrio parahaemolyticus have been reported previously, the simultaneous isolation of two marine vibrios from our case suggests that these organisms may coexist in mixed infections from a common source.

Adult↗
PubMed1997

The lonS gene regulates swarmer cell differentiation of Vibrio parahaemolyticus.

Vibrio parahaemolyticus differentiates from a polarly flagellated, short, rod-shaped cell known as the swimmer to the elongated, hyperflagellated, and multinucleated swarmer cell type when it is grown on a surface. The swarmer is adapted to movement over and colonization of surfaces. To understand the signal transduction mechanism by which the bacterium recognizes surfaces and reprograms gene expression, we isolated a new class of mutants defective in surface sensing. These mutants were constitutive for swarmer cell gene expression, inappropriately expressing high levels of a swarmer cell gene fusion product when grown in liquid. They showed no defect in the swimming motility system, unlike all previously isolated constitutive mutants which have defects in the alternate, polar motility system. The lesions in the majority of the newly isolated mutants were found to be in a gene, lonS, which encodes a polypeptide exhibiting 81% sequence identity to the Escherichia coli Lon protein, an ATP-dependent protease. Upstream sequences preceding the lonS coding region resemble a heat shock promoter, and the homology extends to sequences flanking lonS. The gene order appears to be clpX lonS hupB, like the organization of the E. coli locus. V. parahaemolyticus lonS complemented E. coli lon mutants to restore UV resistance and capsular polysaccharide regulation to that of the wild type. Vibrio lonS mutants were UV sensitive. In addition, when grown in liquid and examined in a light microscope, lonS mutant cells were extremely long and thus resembled swarmer cells harvested from a surface.

ATP-Dependent Proteases↗
PubMed1998

OpaR, a homolog of Vibrio harveyi LuxR, controls opacity of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is an organism well adapted to communal life on surfaces. When grown on a surface or in a viscous layer, the bacterium induces a large gene system and differentiates to swarmer cells capable of movement over and colonization of surfaces. V. parahaemolyticus displays additional phenotypic versatility manifested as variable colony morphology, switching between translucent and opaque colony types. Although not itself luminescent, V. parahaemolyticus produces autoinducer molecules capable of inducing luminescence in Vibrio harveyi. To examine the role of quorum signaling in the lifestyles of V. parahaemolyticus, the functional homolog of the gene encoding the V. harveyi autoinducer-controlled transcriptional regulatory protein LuxR was cloned. Sequence analysis of the clone predicted an open reading frame with a deduced product 96% identical to LuxR. Introduction of the clone carrying the luxR-like locus into V. parahaemolyticus dramatically affected colony morphology, converting a translucent strain to an opaque one. When the coding sequence for the luxR homolog was placed under the control of the Ptac promoter, conversion to the opaque phenotype became inducible by isopropyl-beta-D-thiogalactopyranoside. Allelic disruption of the luxR-like gene on the chromosome of an opaque strain produced a translucent strain proficient in swarming ability. Primer extension mapping demonstrated opaR transcription in opaque but not translucent cell types. It is postulated that this gene, which has been named opaR, encodes a transcription factor controlling cell type. The underlying genetic basis for opaque-translucent variation may be the consequence of a genomic alteration detected in the opaR locus of opaque and translucent strains.

Bacterial Proteins↗
PubMed2005

Physiological roles of three Na+/H+ antiporters in the halophilic bacterium Vibrio parahaemolyticus.

Vibrio parahaemolyticus mutants lacking three Na+/H+ antiporters (NhaA, NhaB, NhaD) were constructed. The DeltanhaA strains showed significantly higher sensitivity to LiCl regarding their growth compared to the parental strain. The DeltanhaA and DeltanhaB strains exhibited higher sensitivities to LiCl. The mutant XACabd lacking all of the three antiporters could not grow in the presence of 500 mM LiCl at pH 7.0, or 50 mM at pH 8.5. The XACabd mutant was also sensitive to 1.0 M NaCl at pH 8.5. These results suggest that Na+/H+ antiporters, especially NhaA, are responsible for resistance to LiCl and to high concentrations of NaCl. Reduced Na+/H+ and Li+/H+ antiport activities were observed with everted membrane vesicles of DeltanhaB strains. However, Li+/H+ antiport activities of DeltanhaB strains were two times higher than those of DeltanhaA strains when cells were cultured at pH 8.5. It seems that expression of nhaA and nhaB is dependent on medium pH to some extent. In addition, HQNO (2-heptyl-4-hydroxyquinoline N-oxide), which is a potent inhibitor of the respiratory Na+ pump, inhibited growth of XACabd, but not of the wild type strain. Moreover, survival rate of XACabd under hypoosmotic stress was lower than that of wild type strain. It is likely that the Na+/H+ antiporters are involved in osmoregulation under hypoosmotic stress. Based on these findings, we propose that the Na+/H+ antiporters cooperate with the respiratory Na+ pump in ionic homeostasis in V. parahaemolyticus.

Lithium↗
PubMed2004

Functional characterization of two type III secretion systems of Vibrio parahaemolyticus.

Vibrio parahaemolyticus, a gram-negative marine bacterium, is a worldwide cause of food-borne gastroenteritis. Recent genome sequencing of the clinical V. parahaemolyticus strain RIMD2210633 identified two sets of genes for the type III secretion system (TTSS), TTSS1 and TTSS2. Here, we constructed a series of mutant strains from RIMD2210633 to determine whether the two putative TTSS apparatus are functional. The cytotoxic activity of mutant strains having a deletion in one of the TTSS1 genes was significantly decreased compared with that of the parent and TTSS2-related mutant strains. In an enterotoxicity assay with the rabbit ileal loop test, intestinal fluid accumulation was diminished by deletion of the TTSS2-related genes while TTSS1-related mutants caused a level of fluid accumulation similar to that of the parent. VopD, a protein encoded in the proximity of the TTSS1 region and a homologue of the Yersinia YopD, was secreted in a TTSS1-dependent manner. In contrast, VopP, which is encoded by a pathogenicity island on chromosome 2 and is homologous to the Yersinia YopP, was secreted via the TTSS2 pathway. These results provide evidence that V. parahaemolyticus TTSSs function as secretion systems and may have a role in the pathogenicity of the organism. This is the first report of functional TTSSs in Vibrio species. The presence of TTSS apparatus gene homologues was demonstrated in other vibrios, such as Vibrio alginolyticus, Vibrio harveyi, and Vibrio tubiashii, suggesting that some other vibrios also contain TTSS and that the TTSS has a role in protein secretion in those organisms during interaction with eukaryotic cells.

Animals↗
PubMed1999

The multiple identities of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a ubiquitous marine bacterium and human pathogen. The organism possesses multiple cell types appropriate for life under different circumstances. The swimmer cell, with a single polar flagellum, is adapted to life in liquid environments. The polar flagellum is powered by the sodium motive force and can propel the bacterium at fast speeds. The swarmer cell, propelled by many proton-powered lateral flagella, can move through highly viscous environments, colonize surfaces, and form multicellular communities which sometimes display highly periodic architecture. Signals that induce differentiation to the surface-adapted cell type are both physical and chemical in nature. Surface-induced gene expression may aid survival, whether attached to inanimate surfaces or in a host organism. Genetic rearrangements create additional phenotypic versatility, which is manifested as variable opaque and translucent colony morphotypes. Discovery that a LuxR homolog controls the opaque cell type implicates intercellular signaling as an additional survival strategy. The alternating identities of V. parahaemolyticus may play important roles in attachment and detachment, how bacterial populations adapt to growth on surfaces, form structured communities, and develop biofilms.

Animals↗
PubMed1996

Utilization of iron sources and its possible roles in the pathogenesis of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is an important enteropathogen in Japan, Taiwan and other coastal regions. The influence of the regulation of iron on the pathogenesis of this pathogen has not been well characterized. The growth of pathogenic and non-pathogenic strains of V. parahaemolyticus on iron-limited agar plates was stimulated by ferritin, lactoferrin and transferrin at 30 microM, and also by hemin, hemoglobin and ferric ammonium citrate at 100 microM. Spontaneous iron-utilizing mutant strains (mutants) were derived from a clinical strain, ST550. Compared with the parent strain, lowered virulence was demonstrated for these mutants, as assayed by adult mouse and suckling mouse models. The in vivo growth and enterotoxigenicity of these mutants were also lower in the suckling mice. Adherence of the mutants to excised mouse intestine was lower as demonstrated by scanning electron microscopy. The iron-regulated outer membrane protein profile also changed in selected mutants. These results indicate that iron-regulated outer membrane proteins and other unknown factors associated with iron utilization may have profound influences, besides iron acquisition, on the pathogenesis of V. parahaemolyticus.

Animals↗
PubMed1993

Cation flux studies of the lesion induced in human erythrocyte membranes by the thermostable direct hemolysin of Vibrio parahaemolyticus.

Vibrio parahaemolyticus, an important agent of seafood-borne gastroenteritis, expresses several putative virulence factors that could account for the disease symptoms of infected humans, namely, diarrhea, nausea, and abdominal cramps. The pathogenicity of V. parahaemolyticus correlates well with the Kanagawa phenomenon (the hemolytic ability of strains grown on Wagatsuma blood agar), implicating the thermostable direct hemolysin (TDH) as the predominant toxin responsible for pathogenicity. TDH-induced hemolysis could be inhibited by the addition of the osmolyte sorbitol to the extracellular solution, supporting the hypothesis that hemolysis occurs through colloid osmosis secondary to an increase in the cation permeability of the membrane. The effect of TDH on cation permeability was investigated by measuring K+ (congener, 86Rb+) influx into human erythrocytes in which the endogenous cation transporters had been blocked (by use of ouabain, bumetanide, and nitrendipine). TDH increased K+ influx into these cells; this increase was rapid in onset and constant in magnitude, suggesting a direct action by TDH on the membrane. The kinetics of leak generation were examined; the relationship between counts accumulated and hematocrit indicated that the TDH-induced lesion is multihit in nature. TDH-induced K+ influx was sensitive to Zn2+. Time courses of hemolysis in isosmotic solutions of monovalent cation chlorides were used to obtain the selectivity series for the TDH-induced leak: Cs+ > Li+ > K+ > Rb+ > Na+. Both the Zn2+ sensitivity and this selectivity series were obtained for crude culture supernatants, suggesting that TDH is the predominant leak-inducing agent. Thus, we have identified several features of the TDH-induced leak likely to be important in the diarrhetic action of V. parahaemolyticus in the human intestine.

Adult↗
PubMed1996

Purification, characterization, and pathogenicity of urease produced by Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a halophilic bacterium associated with gastroenteritis and traveller's diarrhea. The urease-positive, Kanagawa-negative V. parahaemolyticus had been isolated from patients and the environment in the Pacific Northwest, first reported by Kelly et al. (5). Recently, we purified the urease produced by a clinical isolated of V. parahaemolyticus, and its characterization and pathogenicity has been studied. The urease isolation procedure included a water extraction and anion exchange chromatography. The molecular weight for the native enzyme was 275 KDa, and the three subunits of 85 KDa, 59 KDa, and 33 KDa were determined. The isoelectric focusing of urease was 5.2. The purified urease also can cause intestinal fluid accumulation and demonstrate a positive result in the suckling mouse test. These results suggested that the urease produced by V. parahaemolyticus may be another important indicator for the pathogenesis of the bacteria.

Animals↗
PubMed1992

Effects of iron limitation on production of a siderophore, outer membrane proteins, and hemolysin and on hydrophobicity, cell adherence, and lethality for mice of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is one of the most important enteropathogens in Taiwan, Japan, and other coastal regions. The pathogenesis of V. parahaemolyticus disease is not clearly understood. The expression of some factors by V. parahaemolyticus in iron-rich and iron-limited media was analyzed. In the clinical hemolytic strains, the production of a siderophore, two outer membrane proteins (77 and 80 kDa), and thermostable direct hemolysin was significantly enhanced in iron-limited culture, and hemolytic activities, cell hydrophobicity, HEp-2 cell adherence, and lethality for mice were also enhanced. The environmental nonhemolytic strain CCRC12958 that was cultured in iron-limited medium exhibited lethal activity for mice, and other factors except hemolysis were also enhanced like the responses of clinical strains were. These results suggested that a virulent factor(s) of V. parahaemolyticus may be induced or enhanced under iron-limited conditions. The iron-regulated factors reported in this paper may be important in the pathogenesis of V. parahaemolyticus disease.

Animals↗
PubMed2004

Cloning, sequencing and expression of GroEL-like protein gene of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is an important enteropathogen in regions where much seafood is consumed. Substantial quantity of GroEL-like protein is produced during the heat shock of V. parahaemolyticus and located in periplasmic and extracellular fractions. In this study, the GroEL-like protein gene of this pathogen was cloned and sequenced and its properties were analyzed. The open reading frame consisted of 1647 bp, encoded a 57.6-kDa GroEL-like protein of 548 amino acids. The amino acid sequence, hydrophobicity and antigenic pattern of V. parahaemolyticus GroEL-like protein were similar to the GroEL proteins of other vibrios, Escherichia coli and Salmonella enterica Typhi. The groEL-like gene of V. parahaemolyticus was cloned into the pQE-30 expression vector and the expression of a His-tagged GroEL-like protein was rapidly induced by isopropyl-beta-D-thiogalactopyranoside, largely in an insoluble form. The results of this study facilitate the study of the functions of the GroEL-like protein of V. parahaemolyticus.

Amino Acid Sequence↗
PubMed1980

Sugar composition of O-antigenic lipopolysaccharides isolated from Vibrio parahaemolyticus.

Vibrio parahaemolyticus, a causative bacterium of food poisoning unique for its particular primary association with sea products, is now divided serologically into 11 or 12 O-forms based on agglutination and agglutinin-absorption tests. We determined the sugar composition of the somatic O-antigens, i.e., lipopolysaccharides (LPS), of representative strains of each O-form. Of particular interest is the absence of evidence for the presence of 2-keto-3-deoxy-octonic acid (KDO), a regular sugar component of gram-negative bacterial LPS, in any LPS examined, with the exception of 06. Furthermore, 07 and 012 LPS contained a KDO-like compound that is, however, not identical with KDO. Glucose, glucosamine, and L-glycero-D-mannoheptose were found as common sugar constituents. Three unidentified amino sugars, designated here as P1, P2, and P3, were found. Various combinations of each of these unidentified amino sugars, and of galactose, fucose, arabinose, D-glyucero-D-mannoheptose, galactosamine, KDO, and the KDO-like substance were detected in accordance with the O-form of LPS. On the basis of the sugar composition, LPS of the 12 O-forms of V. parahaemolyticus can be classified into nine chemotypes, because 03, 05, and 011 LPS belong to the same chemotype and 07 and 012 to another chemotype.

Antigens, Bacterial↗
PubMed2004

Biochemical and virulence characterization of viable but nonculturable cells of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a common foodborne pathogen frequently causing outbreaks in summer. Maintenance of virulence by the viable but nonculturable (VBNC) state of this pathogen would allow its threat to human health to persist. This study reports on the change in virulence and concomitant changes in activity of two enzymes and fatty acid profiles when V. parahaemolyticus ST550 entered the VBNC state in the modified Morita mineral salt-0.5% NaCl medium incubated at 4 degrees C. The major change in fatty acid composition occurred in the first week, with a rapid increase in C15:0 fatty acid and saturated/unsaturated ratio while a rapid decrease in C16:1 was observed. The activity level of the inducible protective enzyme superoxide dismutase became undetectable in the VBNC state, whereas that of constitutive glucose-6-phosphate dehydrogenase did not change in either the exponential phase or the VBNC state. Cytotoxicity against HEp-2 cells and a suckling mouse assay showed that virulence was lowered in the VBNC state compared with exponential-phase cells. Longer incubation times were required by the VBNC cells to achieve the same level of virulence as seen in exponential-phase cells. Culturable cells were recovered on selective agar medium from the VBNC cultures injected into suckling mice, probably as the result of in vivo resuscitation. Results of this study add to our understanding of the biochemical and physiological changes that have not been reported when V. parahaemolyticus enters into the VBNC state.

Animals↗
PubMed2005

Genetic determinants of biofilm development of opaque and translucent Vibrio parahaemolyticus.

Vibrio parahaemolyticus isolates display variation in colony morphology, alternating between opaque (OP) and translucent (TR) cell types. Phase variation is the consequence of genetic alterations in the locus encoding the quorum sensing output regulator OpaR. Here, we show that both cell types form stable, but distinguishable biofilms that differ with respect to attachment and detachment profiles to polystyrene, pellicle formation and stability at the air/medium interface, and submerged biofilm architecture and dispersion at a solid/liquid interface. The pellicle, which is a cohesive mat of cells, was exploited to identify mutants having altered or defective biofilm formation. Transposon insertion mutants were obtained with defects in genes affecting multiple cell surface characteristics, including extracellular polysaccharide, mannose-sensitive haemagglutinin type 4 pili and polar (but not lateral) flagella. Other insertions disrupted genes coding for potential secreted proteins or transporters of secreted proteins, specifically haemolysin co-regulated protein and an RTX toxin-like membrane fusion transporter, as well as potential modifiers of cell surface molecules (nagAC operon). The pellicle screen also identified mutants with lesions in regulatory genes encoding H-NS, a CsgD-like repressor and an AraC-like protein. This work initiates the characterization of V. parahaemolyticus biofilm formation in the OP and TR cell types and identifies a diverse repertoire of cell surface elements that participate in determining multicellular architecture.

Bacterial Proteins↗