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PubMed1976

[The dehydrogenation of butandiol by "vibrio parahaemolyticus" and other vibrios (author's transl)].

The butandiol dehydrogenase was examined in 64 vibrios isolated in Togo 1970/71. This enzyme was found in all of the not halophilic strains and in the species Vibrio alginolyticus, whereas the species V. parahaemolyticus exhibited as well dehydrating as not dehydrating strains. The butandiol dehydrogenase could be a good marker for epidemiologic analysis in characterizing V. parahaemolyticus strains isolated from different sources, especially if no antisera are available.

Alcohol Oxidoreductases↗
PubMed1991

Analysis of the gene of Vibrio hollisae encoding the hemolysin similar to the thermostable direct hemolysin of Vibrio parahaemolyticus.

The gene (designated as Vh-tdh) of Vibrio hollisae 9041 encoding a hemolysin similar to the thermostable direct hemolysin (TDH) of V. parahaemolyticus contained a 567-base-pair open reading frame (ORF), which was 93.3-93.5% homologous to those of the tdh genes of V. parahaemolyticus, V. cholerae non-01, and V. mimicus encoding TDH or similar hemolysins. Comparative analysis of the nucleotide sequence containing the Vh-tdh ORF with published nucleotide and amino acid sequences suggested that the Vh-tdh gene and other tdh genes diverged from a common ancestral gene, that the divergence was closely associated with the evolutionary divergence of V. hollisae from other species of genus Vibrio, and that strain-to-strain variation of the Vh-tdh gene exists in V. hollisae.

Amino Acid Sequence↗
PubMed1991

Similarity of the tdh gene-bearing plasmids of Vibrio cholerae non-O1 and Vibrio parahaemolyticus.

The gene encoding a hemolysin similar to the thermostable direct hemolysin (TDH) of Vibrio parahaemolyticus was previously cloned from a plasmid of Vibrio cholerae non-O1. The gene (designated as NAG-tdh) was subcloned and its nucleotide sequence was determined and compared with reported sequences of the four tdh gene copies encoding TDH, of which three were cloned from the chromosome and one was cloned from a plasmid of V. parahaemolyticus. In the coding region, the NAG-tdh gene had 100% homology with the plasmid-borne tdh gene (tdh4) whereas the NAG-tdh gene was 96.7-98.6% homologous to the three chromosomal tdh genes. The sequences of the NAG-tdh and tdh4 genes were nearly identical in the further upstream and downstream regions. The entire plasmids carrying the two tdh genes were found to be highly homologous when compared by restriction endonuclease and Southern blot analyses. The results suggest that the tdh gene has been transferred between V. cholerae non-O1 and V. parahaemolyticus by a plasmid, directly or indirectly, and that the nucleotide sequences of the tdh gene-bearing plasmids have undergone minor base changes in the respective genetic backgrounds.

Amino Acid Sequence↗
PubMed2004

The small RNA chaperone Hfq and multiple small RNAs control quorum sensing in Vibrio harveyi and Vibrio cholerae.

Quorum-sensing bacteria communicate with extracellular signal molecules called autoinducers. This process allows community-wide synchronization of gene expression. A screen for additional components of the Vibrio harveyi and Vibrio cholerae quorum-sensing circuits revealed the protein Hfq. Hfq mediates interactions between small, regulatory RNAs (sRNAs) and specific messenger RNA (mRNA) targets. These interactions typically alter the stability of the target transcripts. We show that Hfq mediates the destabilization of the mRNA encoding the quorum-sensing master regulators LuxR (V. harveyi) and HapR (V. cholerae), implicating an sRNA in the circuit. Using a bioinformatics approach to identify putative sRNAs, we identified four candidate sRNAs in V. cholerae. The simultaneous deletion of all four sRNAs is required to stabilize hapR mRNA. We propose that Hfq, together with these sRNAs, creates an ultrasensitive regulatory switch that controls the critical transition into the high cell density, quorum-sensing mode.

Amino Acid Sequence↗
PubMed2005

A variant type of Vibrio cholerae SXT element in a multidrug-resistant strain of Vibrio fluvialis.

Vibrio fluvialis strain H-08942 was isolated from an infant aged 6 months who was suffering from cholera-like diarrhea in India. This strain showed the typical multidrug-resistance phenotype of an SXT element. It was resistant to sulfamethoxazole (Su), trimethoprim (Tm), chloramphenicol (Cm) and streptomycin (Sm), in addition to other antibiotics such as ampicillin (Am), furazolidone (Fz), nalidixic acid (Na), and gentamicin (Gm). The SXT element is a Vibrio cholerae-derived integrative and conjugative element (ICE) that has also been referred to as a conjugative transposon. Our goal was to find a relationship between these resistant phenotypes and the presence of the SXT element in this unique strain. By using PCR, we detected the antibiotic resistance genes, the integrase gene and the attP attachment site of SXT element. Cloning and DNA sequencing results showed that both the SXT integrase gene and its attP site of V. fluvialis were similar but not identical to those of V. cholerae. The SXT integrase gene of V. fluvialis has a 99% identity to that of V. cholerae, and the attP site of SXT of V. fluvialis is variant and shorter (641 bp) than that of V. cholerae (785 bp). It was possible for the SXT of V. fluvialis to be transferred by conjugation to a laboratory strain of Escherichia coli. Here, we report the detection of a variant SXT element in species other than V. cholerae, with molecular characterization and analysis of its integrase gene and its attP site.

Conjugation, Genetic↗
PubMed1982

Illness caused by Vibrio damsela and Vibrio hollisae.

Vibrio damsela was isolated from six wound infections in otherwise healthy persons. In five of the six cases the wounds were known to have been exposed to salt or brackish water at the time of the injury. Vibrio hollisae was isolated from an index stool culture in nine cases in which no other enteric pathogen was identified. All nine patients had diarrhoea and abdominal pain; one patient had bloody diarrhoea. Six of the nine patients were known to have eaten raw seafood in the five days before they became ill. These data suggest that both V. damsela and V. hollisae can produce diseases with distinct clinical and epidemiological characteristics.

Abdomen↗
PubMed2005

Isolation of Vibrio parahaemolyticus and Vibrio vulnificus from wild aquatic birds in Japan.

Vibrio parahaemolyticus and Vibrio vulnificus were isolated from faecal samples of wild aquatic birds in winter. Although V. parahaemolyticus and V. vulnificus were present in low numbers in seawater in the area where the faecal samples of the birds were collected, the pathogens were isolated from the faeces of the birds. This study demonstrates that wild aquatic birds are a vehicle for V. parahaemolyticus and V. vulnificus to survive in winter.

Animals↗
PubMed2003

Molecular characterisation and antimicrobial resistance of Vibrio vulnificus and Vibrio alginolyticus isolated from mussels (Mytilus galloprovincialis).

Twelve Vibrio vulnificus biotype 1 and 11 Vibrio alginolyticus isolated from mussels in Italy were analysed by antimicrobial resistance, plasmid profiles, random amplification of polymorphic DNA (RAPD), and single enzyme amplified fragment length polymorphism (sAFLP). Plasmid DNA was detected in three V. vulnificus and four V. alginolyticus cultures. All isolates were resistant to at least two antimicrobial agents: all isolates were resistant to ampicillin, carbenicillin and streptomycin, except one V. alginolyticus which was sensitive to carbenicillin and two V. alginolyticus which were sensitive to streptomycin. No association was detected between the presence of plasmid DNA and antimicrobial resistance. Seven of the twelve V. vulnificus and two of the eleven V. alginolyticus cultures were susceptible to the 10 microg of the vibriostatic compound O/129; all cultures were susceptible to the 150 microg of O/129. Both RAPD and sAFLP was found to be reproducible. Ten sAFLP and seven RAPD profiles were detected amongst the 12 V. vulnificus cultures: three cultures were identified as indistinguishable by both methods. RAPD and sAFLP analysis of V. alginolyticus generated nine and seven profiles respectively, and these two methods were independent. These results demonstrate extreme variability of V. vulnificus and V. alginolyticus isolated from mussels, and both RAPD and sAFLP provided information on intraspecific differences which will be useful for molecular epidemiological or ecological studies. A combination of methods gave optimal discrimination, although a single method could provide sufficient information to characterise V. vulnificus isolates.

Animals↗
PubMed2002

Vibrio trachuri Iwamoto et al. 1995 is a junior synonym of Vibrio harveyi (Johnson and Shunk 1936) Baumann et al. 1981.

The taxonomic position of Vibrio trachuri was examined through a polyphasic approach using 16S rDNA sequencing, fluorescent amplified fragment length polymorphisms (FAFLP), DNA-DNA hybridization experiments, G+C content of DNA and phenotypical tests. Phylogenetic analysis showed that Vibrio harveyi is the closest neighbour of V. trachuri, sharing about 98.8% similarity in the 16S rDNA gene. Moreover, numerical analysis of the FAFLP patterns revealed that both species have highly related genomes, sharing 55% pattern similarity. DNA-DNA hybridization experiments and G+C content measurements reinforced these results, since V. trachuri and V. harveyi had at least 74% DNA similarity and 44.5-45.2 mol % G+C. Phenotypical features of both species were also very similar, except that V. trachuri utilized itaconic acid, whereas V. harveyi did not. Therefore, it is proposed that the species V. trachuri should be reclassified as V. harveyi.

Animals↗
PubMed1991

Two generalized transducing phages in Vibrio parahaemolyticus and Vibrio alginolyticus.

Two bacteriophages named phi VP253 and phi VP143 isolated after ultraviolet induction from lysogenic strains of Vibrio parahaemolyticus have been shown to be generalized transducing phages. So far, seven different auxotrophic markers of a V. parahaemolyticus strain could be transduced at the frequencies ranging from 2.2 x 10(-7) to 7.5 x 10(-5) per infected cell at the m.o.i. of approximately 1.0. The phage phi VP143, but not phi VP253, lysed 20 of the 28 strains of V. alginolyticus and the occurrence of generalized transduction by this phage in this Vibrio species has been confirmed. Molecular size of the genomes of both phages were estimated to be approximately 48 kb as judged from electrophoretic mobilities of the DNAs digested with HindIII endonuclease. The results and similarity of the two phages in morphology and other properties suggest very close relatedness of the phages.

Animals↗
PubMed1999

Disappearance of glyceraldehyde 3-phosphate dehydrogenase from erythrocyte membrane by hemolysis with thermostable direct hemolysin of Vibrio parahaemolyticus or Vibrio cholerae El Tor hemolysin.

It is believed that the thermostable direct hemolysin (TDH) of Vibrio parahaemolyticus and El Tor hemolysin (ETH) of Vibrio cholerae damage erythrocytes and other cells by acting as pore-forming toxins. In this study, we found that a protein band with a molecular weight of 37,000 daltons specifically disappeared from erythrocyte membrane after hemolysis by TDH and ETH, but not after hypotonic hemolysis. The 37 kDa band was identified as glyceraldehyde 3-phosphate dehydrogenase (G3PD), a glycolytic enzyme, based on N-terminal 14 amino acid sequencing. The role of G3PD in hemolysis is discussed.

Bacterial Proteins↗
PubMed1983

Enterotoxin production by Vibrio cholerae and Vibrio mimicus grown in continuous culture with microbial cell recycle.

We have examined the effect of complete cell recycle on the production of cholera toxin (CT) by Vibrio cholerae and CT-like toxin by Vibrio mimicus in continuous culture fermentations. Complete cell recycle was obtained by filtering culture fluids through Amicon hollow fibers with an exclusion limit of 100,000 daltons (H1P100-20) and returning the concentrated cell slurry to the fermentor. A single 1-liter laboratory fermentor system modified with this recycle loop was capable of producing over 20 liters of cell-free culture filtrate per day. Toxin production in this system was compared with yields obtained in traditional continuous cultures and in shake flask cultures. Yields of CT from V. cholerae 569B in the recycle fermentor were highest at the highest dilution rate employed (1.0 vol/vol per h). The use of complete cell recycle dramatically increased yields over those obtained in continuous culture and equaled those obtained in shake flasks. The concentration of CT in the filtrate was slightly less than half of that measured in culture fluids sampled at the same time. Similarly, V. mimicus 61892 grown in the presence of 50 micrograms of lincomycin per ml produced 280 ng of CT per ml in the recycle fermentor, compared with 210 ng/ml in shake flasks under optimal conditions. The sterile filtrate from this fermentation contained 110 ng/ml.

Cholera Toxin↗
PubMed1999

Analysis of 16S-23S rRNA intergenic spacer regions of Vibrio cholerae and Vibrio mimicus.

Vibrio cholerae identification based on molecular sequence data has been hampered by a lack of sequence variation from the closely related Vibrio mimicus. The two species share many genes coding for proteins, such as ctxAB, and show almost identical 16S DNA coding for rRNA (rDNA) sequences. Primers targeting conserved sequences flanking the 3' end of the 16S and the 5' end of the 23S rDNAs were used to amplify the 16S-23S rRNA intergenic spacer regions of V. cholerae and V. mimicus. Two major (ca. 580 and 500 bp) and one minor (ca. 750 bp) amplicons were consistently generated for both species, and their sequences were determined. The largest fragment contains three tRNA genes (tDNAs) coding for tRNAGlu, tRNALys, and tRNAVal, which has not previously been found in bacteria examined to date. The 580-bp amplicon contained tDNAIle and tDNAAla, whereas the 500-bp fragment had single tDNA coding either tRNAGlu or tRNAAla. Little variation, i.e., 0 to 0.4%, was found among V. cholerae O1 classical, O1 El Tor, and O139 epidemic strains. Slightly more variation was found against the non-O1/non-O139 serotypes (ca. 1% difference) and V. mimicus (2 to 3% difference). A pair of oligonucleotide primers were designed, based on the region differentiating all of V. cholerae strains from V. mimicus. The PCR system developed was subsequently evaluated by using representatives of V. cholerae from environmental and clinical sources, and of other taxa, including V. mimicus. This study provides the first molecular tool for identifying the species V. cholerae.

Base Sequence↗
PubMed1983

Production of cholera toxin-like toxin by Vibrio mimicus and non-O1 Vibrio cholerae: batch culture conditions for optimum yields and isolation of hypertoxigenic lincomycin-resistant mutants.

Vibrio mimicus 61892, isolated in 1977 from a case of watery diarrhea in Bangladesh, produces an enterotoxin which possesses activity in Y-1 mouse adrenal cells and in rabbit ileal loops which is identical to the prototype cholera toxin (CT) produced by Vibrio cholerae 569B. The neutralization of the adrenal cell activity of 61892 toxin and 569B CT by homologous and heterologous antisera generates parallel titration curves which show complete neutralization in all cases. Paired titrations in the ganglioside GM1 enzyme-linked immunosorbent assay (using either CT or Escherichia coli heat-labile toxin antitoxin) of both toxins indicates that 61892 toxin is antigenically indistinguishable from 569B CT. The specific activity of the two toxins in the rabbit ileal loop is virtually identical. Batch culture production of CT-like toxin and CT by isolates of V. mimicus and different biotypes of V. cholerae was found to be highest in shake flask cultures of Casamino Acids-yeast extract broth grown at 27 degrees C with vigorous aeration. Incorporation of lincomycin into the growth medium at a concentration of 50 micrograms/ml increased yields from wild-type strains. Dramatically higher yields were obtained when a spontaneous resistance mutant of strain 61892 was grown in the presence of 200 to 300 micrograms of lincomycin per ml. Under these conditions, yields of CT-like toxin were increased by 300- to 500-fold, and the highest yields reached more than 100 micrograms/ml after 44 h of culture. This is substantially higher than that reported in the literature for CT production by any strain of V. cholerae, including hypertoxigenic strain 569B.

Bacterial Toxins↗
PubMed1986

Purification and partial characterization of a non-O1 Vibrio cholerae hemolysin that cross-reacts with thermostable direct hemolysin of Vibrio parahaemolyticus.

A newly identified hemolysin (NAG-rTDH), which is related to the thermostable direct hemolysin (Vp-TDH) of Vibrio parahaemolyticus produced by non-O1 Vibrio cholerae, was studied. NAG-rTDH was purified by successive column chromatographies on DEAE-cellulose and an immunoaffinity column coupled with anti-Vp-TDH immunoglobulin. The molecular weight of NAG-rTDH was estimated as 18,500, similar to that of Vp-TDH, as judged by sodium dodecyl sulfate slab gel electrophoresis, but its charge or molecular shape was different, judging from its electrophoretic mobility. The lytic activities of NAG-rTDH on erythrocytes of most animals were essentially similar to those of Vp-TDH, but that on sheep erythrocytes was different. The hemolytic activity of NAG-rTDH was stable on heating at 100 degrees C for 10 min, as was that of Vp-TDH. Immunological cross-reactivity between NAG-rTDH and Vp-TDH was demonstrated by both the Ouchterlony test and the neutralization test. Thus, we conclude that non-O1 V. cholerae produce a new type of hemolysin that is similar but not identical to the thermostable direct hemolysin of V. parahaemolyticus.

Cross Reactions↗
PubMed1990

The cytolysin gene of Vibrio vulnificus: sequence and relationship to the Vibrio cholerae E1 Tor hemolysin gene.

A cytolysin of ca. 56 kilodaltons has been suggested as a possible virulence factor in Vibrio vulnificus infections. We sequenced the DNA encoding cytolytic activity and found that the sequence contained two open reading frames, vvhA and vvhB. vvhA encoded the structural gene for the cytolysin and contained the N-terminal amino acid sequence previously reported for the protein. Regions of the vvhA gene showed homology to the structural gene for the Vibrio cholerae E1 Tor hemolysin.

Amino Acid Sequence↗