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PubMed2005

Molecular evolution of Vibrio pathogenicity island-2 (VPI-2): mosaic structure among Vibrio cholerae and Vibrio mimicus natural isolates.

Vibrio cholerae is a Gram-negative rod that inhabits the aquatic environment and is the aetiological agent of cholera, a disease that is endemic in much of Southern Asia. The 57.3 kb Vibrio pathogenicity island-2 (VPI-2) is confined predominantly to toxigenic V. cholerae O1 and O139 serogroup isolates and encodes 52 ORFs (VC1758 to VC1809), which include homologues of an integrase (VC1758), a restriction modification system, a sialic acid metabolism gene cluster (VC1773-VC1783), a neuraminidase (VC1784) and a gene cluster that shows homology to Mu phage. In this study, a 14.1 kb region of VPI-2 comprising ORFs VC1773 to VC1787 was identified by PCR and Southern blot analyses in all 17 Vibrio mimicus isolates examined. The VPI-2 region in V. mimicus was inserted adjacent to a serine tRNA similar to VPI-2 in V. cholerae. In 11 of the 17 V. mimicus isolates examined, an additional 5.3 kb region encoding VC1758 and VC1804 to VC1809 was present adjacent to VC1787. The evolutionary history of VPI-2 was reconstructed by comparative analysis of the nanH (VC1784) gene tree with the species gene tree, deduced from the housekeeping gene malate dehydrogenase (mdh), among V. cholerae and V. mimicus isolates. Both gene trees showed an overall congruence; on both gene trees V. cholerae O1 and O139 serogroup isolates clustered together, whereas non-O1/non-O139 serogroup isolates formed separate divergent branches with similar clustering of strains within the branches. One exception was noted: on the mdh gene tree, V. mimicus sequences formed a distinct divergent lineage from V. cholerae sequences; however, on the nanH gene tree, V. mimicus clustered with V. cholerae non-O1/non-O139 isolates, suggesting horizontal transfer of this region between these species.

Bacterial Proteins↗
PubMed2006

Quorum sensing-disrupting brominated furanones protect the gnotobiotic brine shrimp Artemia franciscana from pathogenic Vibrio harveyi, Vibrio campbellii, and Vibrio parahaemolyticus isolates.

Autoinducer 2 (AI-2) quorum sensing was shown before to regulate the virulence of Vibrio harveyi towards the brine shrimp Artemia franciscana. In this study, several different pathogenic V. harveyi, Vibrio campbellii, and Vibrio parahaemolyticus isolates were shown to produce AI-2. Furthermore, disruption of AI-2 quorum sensing by a natural and a synthetic brominated furanone protected gnotobiotic Artemia from the pathogenic isolates in in vivo challenge tests.

Animals↗
PubMed1994

Identification of the genes encoding NAD(P)H-flavin oxidoreductases that are similar in sequence to Escherichia coli Fre in four species of luminous bacteria: Photorhabdus luminescens, Vibrio fischeri, Vibrio harveyi, and Vibrio orientalis.

Genes encoding NAD(P)H-flavin oxidoreductases (flavin reductases) similar in both size and sequence to Fre, the most abundant flavin reductase in Escherichia coli, were identified in four species of luminous bacteria, Photorhabdus luminescens (ATCC 29999), Vibrio fischeri (ATCC 7744), Vibrio harveyi (ATCC 33843), and Vibrio orientalis (ATCC 33934). Nucleotide sequence analysis showed Fre-like flavin reductases in P. luminescens and V. fischeri to consist of 233 and 236 amino acids, respectively. As in E. coli Fre, Fre-like enzymes in luminous bacteria preferably used riboflavin as an electron acceptor when NADPH was used as an electron donor. These enzymes also were good suppliers of reduced flavin mononucleotide (FMNH2) to the bioluminescence reaction. In V. fischeri, the Fre-like enzyme is a minor flavin reductase representing < 10% of the total FMN reductase. That the V. fischeri Fre-like enzyme has no appreciable homology in amino acid sequence to the major flavin reductase in V. fischeri, FRase I, indicates that at least two different types of flavin reductases supply FMNH2 to the luminescence system in V. fischeri. Although Fre-like flavin reductases are highly similar in sequence to luxG gene products (LuxGs), Fre-like flavin reductases and LuxGs appear to constitute two separate groups of flavin-associated proteins.

Amino Acid Sequence↗
PubMed2006

PCR-based identification of Vibrio cholerae and the closely related species Vibrio mimicus using the large chromosomal ori sequence of Vibrio cholerae.

The bacterial chromosomal replication origin (ori) sequences are a highly conserved essential genetic element. In this study, the large chromosomal replication origin sequence of Vibrio cholerae (oriCIVC) has been targeted for identification of the organism, including the biotypes of serogroup O1. The oriCIVC sequence-based PCR assay specifically amplified an 890 bp fragment from all the V. cholerae strains examined. A point mutation in the oriCIVC sequence of the classical biotype of O1 serogroup led to the loss of a BglII site, which was utilized for differentiation from El Tor vibrios. Interestingly, the PCR assay amplified a similarly sized ori segment, designated as oriCIVM, from V. mimicus strains, but failed to produce any amplicon with other strains. Cloning and sequencing of the oriCIVM revealed high sequence similarity (96%) with oriCIVC. The results indicate that V. mimicus is indeed very closely related to V. cholerae. In addition, the BglII restriction fragment length polymorphism (RFLP) between oriCIVM and oriCIVC sequences allowed us to differentiate the two species. The ori sequence-based PCR-RFLP assay developed in this study appears to be a useful method for rapid identification and differentiation of V. cholerae and V. mimicus strains, as well as for the delineation of classical and El Tor biotypes of V. cholerae O1.

Bacterial Typing Techniques↗
PubMed1988

A comparative study of the sugar composition of lipopolysaccharides isolated from Vibrio cholerae, 'Vibrio albensis' and Vibrio metschnikovii.

A comparative study was made of the quantitative sugar composition of lipopolysaccharides (LPS) isolated from Vibrio cholerae (O1 and non O1 groups), 'V. albensis', 'V. proteus' and V. metschnikovii. The amino sugars 4-amino-4,6-dideoxy-D-mannose (perosamine) and 2-amino-2,6-dideoxy-D-glucose (quinovosamine) were present exclusively in LPS isolated from S-form O1 group of V. cholerae regardless of serotype (i.e. Ogawa or Inaba) and biotype (i.e. classical or eltor). Classical O1 group V. cholerae was distinguishable from eltor O1 group V. cholerae on the basis of the fructose content of the LPS: greater than 3% and less than or equal to 1%, respectively. Distinct differences in the sugar composition of LPS were observed between V. cholerae and 'V. albensis', 'V. proteus' and V. metschnikovii.

Carbohydrates↗
PubMed1983

Medium-dependent production of extracellular enterotoxins by non-O-1 Vibrio cholerae, Vibrio mimicus, and Vibrio fluvialis.

Fluid accumulation at 4 h in the intestines of suckling mice enabled us to distinguish non-O-1 Vibrio cholerae, V. mimicus, and V. fluvialis clinical isolates from environmental isolates. Enterotoxin production was culture medium dependent. Filtrates of cultures grown in tryptic soy broth without glucose but with added 0.5% NaCl did not exhibit marked enterotoxin activity in the assay. Culture filtrates of all clinical strains grown in brain heart infusion broth supplemented with 0.5% NaCl induced large amounts of fluid accumulation in mouse intestines. However, most environmental strains grown in brain heart infusion broth amended as described above were unable to induce fluid accumulation. The enterotoxin present in culture filtrates lost activity at 56 degrees C and appeared to be distinct from previously described virulence factors, including the well-described cholera toxin. The new enterotoxin could represent an important virulence mechanism common to all three species.

Culture Media↗
PubMed1989

Sugar composition of the polysaccharide portion of lipopolysaccharides of Vibrio fluvialis, Vibrio vulnificus, and Vibrio mimicus.

A chemotaxonomic study was carried out on Vibrio fluvialis and V. vulnificus on the basis of the sugar composition of the polysaccharide portion of their lipopolysaccharides (LPS). A previously developed rapid method of preparing samples for compositional sugar analysis was employed. Nineteen O-serogroups of V. fluvialis were divided into 14 chemotypes while seven O-serogroups of V. vulnifucus were divided also into seven chemotypes since the polysaccharide portion of LPS of each serogroup has a different sugar composition from that of the other serogroups. Close similarities in the sugar composition of the same portion were demonstrated between serologically cross-reacting non-O1 group V. cholerae and V. fluvialis, and non-O1 V. cholerae and V. mimicus.

Antigens, Bacterial↗
PubMed1994

Sequence determination of rRNA genes of pathogenic Vibrio species and whole-cell identification of Vibrio vulnificus with rRNA-targeted oligonucleotide probes.

A comparative analysis of seven new 16S rRNA gene sequences of pathogenic Vibrio species with previously published vibrio sequences confirmed that Vibrio vulnificus represents a group that is not closely related to the core organisms of the genus Vibrio. In addition, we found that V. vulnificus, Listonella (Vibrio) anguillarum and Vibrio diazotrophicus branch off separately from the core group. A comparison of the 16S rRNA gene sequences of V. vulnificus strains belonging to biotypes 1 and 2 revealed that the sequences of all but four biotype 1 strains were identical to each other but slightly different (17 bases) from the sequences of the rest of the V. vulnificus strains investigated. In addition, the sequences of variable regions of the 23S rRNA genes of Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio cholerae, and V. vulnificus C7184 and TW1 were determined, aligned, and compared with all available bacterial 23S rRNA sequences in order to search for specific target sites. As a result, four oligonucleotide probes specific for V. vulnificus were synthesized, and the specificities of these probes were evaluated by dot blot hybridization to membrane-bound RNAs from 21 V. vulnificus strains, 13 strains belonging to other Vibrio species, 61 strains belonging to species that are members of the alpha, beta, and gamma subclasses of the Proteobacteria, and 3 eucaryotic microorganisms. Two probes hybridized with all of the V. vulnificus strains tested, and the other two probes distinguished V. vulnificus biotype 1 strains from all other organisms. In situ identification of V. vulnificus by using tetramethylrhodamine- or fluorescein-labelled oligonucleotides is now possible.

Bacterial Typing Techniques↗
PubMed1975

Vibrio cholerae flagellar antigens: a serodiagnostic test, functional implications of H-reactivity and taxonomic importance of cross-reactions within the Vibrio genus.

Serodiagnostic tests for all serotypes of Vibrio cholerae using H-antisera were investigated. Activity motile cell lines of 155 stock and international reference cultures of human, animal, fish, and halophilic Vibrios, Aeromonas, Comomonas, Pseudomonas, Salmonella, and Escherichia were investigated. Without exception, all cholera vibrios (including the NAG serotypes) reacted with H sera. Positive reactions were obtained specifically (a) within 2 hrs at 52 degrees C in the tube test using thick formalized suspensions and H antisera at optimal proportion titre and (b) within 30 sec by slide agglutination of fresh cultures. The other vibrios investigated reacted similarly with their homologous H antisera. 2. The rapid diagnostic techniques of fluorescent antibody labeling or immobilization were unsuccessful, V. cholerae flagella being refractive to H sera in these tests. V. cholerae was, however, sensitive in a type-specific manner to O antisera. These and related observations suggest that O antigen has a functional role in Vibrio motility. 3. Interspecies H cross-reactions between V. cholerae and fish and animal vibrios which correlated with bacteriologic similarity, were demonstrated. O antigens of these vibrios were strain specific. Cross-absorption analysis indicated that the H antigens of vibrios were characteristic and homogenous within the species, and therefore a potentially important taxonomic criterion of Vibrio species.

Adsorption↗
PubMed2000

Phenotypic and genotypic characterization of Vibrio viscosus sp. nov. and Vibrio wodanis sp. nov. isolated from Atlantic salmon (Salmo salar) with 'winter ulcer'.

Two groups of Vibrio strains isolated from Atlantic salmon with 'winter ulcer' were characterized phenotypically and genotypically. The data obtained indicated that each of the two groups represented a new species in the genus Vibrio. The names Vibrio viscosus sp. nov. [type strain NVI 88/478T (= NCIMB 13584T)] and Vibrio wodanis sp. nov. [type strain NVI 88/441T (= NCIMB 13582T)] are proposed for the new species. V. viscosus strains exhibited a similar total DNA RFLP pattern and a similar plasmid DNA profile. DNA relatedness (hydroxyapatite method) of the V. viscosus type strain to nine other V. viscosus strains was 81-93% at 60 degrees C. Divergence within related sequences was 0.0-1.5% and relatedness at 75 degrees C was 74-100%. V. wodanis strains exhibited marked heterogeneity on the basis of RFLP analysis and plasmid profiles. DNA relatedness of the V. wodanis type strain to 10 other V. wodanis strains was 66-94% at 60 degrees C. Divergence within related sequences was 0.0-1.5% and relatedness at 75 degrees C was 55-97%. Relatedness between V. viscosus and V. wodanis type strains was approximately 20%. Among other Vibrio species, the closest relative of V. viscosus was Vibrio marinus (ATCC 15381T) (43% relatedness at 60 degrees C) and that of V. wodanis was Vibrio logei (ATCC 15382) (57% relatedness at 60 degrees C). These same pairs were the closest phenotypic relatives. DNA sequence analysis of the 16S rRNA gene of V. viscosus indicated an intimate relationship to V. marinus. A total evaluation of the results, however, supports V. viscosus to be a separate species in the genus Vibrio. The analysis of the sequence of the 16S rRNA gene of V. wodanis supports that V. logei (ATCC 15382) was the most related species. Ability to degrade casein, oxidative production of acid from trehalose and production of lysine decarboxylase are important biochemical tests that will differentiate between V. viscosus, V. wodanis, V. marinus (ATCC 15381T) and V. logei (ATCC 15382).

Animals↗
PubMed1978

Incidence of Vibrio parahaemolyticus bacteriophages and other Vibrio bacteriophages in marine samples.

Vibrio bacteriophages were isolated by enrichment from 177 of 643 samples of marine molluscan shellfish, crustaceans, seawater, and sediments. The predominant bacteriophage types isolated were specific for some strains of Vibrio parahaemolyticus. A high frequency of phage isolations was also observed with strains of agar-digesting vibrios (21 of 56) and psychrophilic vibrios (14 of 72) that were originally isolated from non-shellfish growing areas. No bacteriophages were isolated against V. alginolyticus and only rarely for V. anguillarum even though these were the two most abundant species found in near-shore environments. No V. cholerae phages were isolated. It was also determined from quantitative studies on the Pacific oyster (Crassostrea gigas) obtained from two environments in Washington and Oregon that the titers of V. parahaemolyticus bacteriophages increased with increasing seasonal water temperatures and that this was proportional to the increase in numbers of mesophilic vibrios and not with the incidence of V. parahaemolyticus. Titers of V. parahaemolyticus bacteriophages occasionally exceeded 10(6) per g of oyster during the summer months. Specific V. parahaemolyticus bacteriophages were also isolated from market seafoods and other marine samples that originated in cold environments where no mesophilic vibrios are expected to be found. The possibility that V. parahaemolyticus bacteriophages originate from Vibrio spp. other than V. parahaemolyticus and the role of these bacteriophages in the ecology of marine vibrios are discussed.

Animals↗