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PubMed2002

The inherent characteristics and DNA polymorphism of Vibrio cholerae and other vibrios.

OBJECTIVE: To investigate the inherent characteristics of Vibrio cholerae (V. cholerae) and other vibrios and their relationship. METHODS: Polymerase chain reaction (PCR), DNA sequence analysis, randomly amplified polymorphic DNA (RAPD) analysis and average linkage cluster analysis were used to study 3 isolates of V. cholerae strains O139, three isolates O1 biotype El Tor, four isolates O1 biotype classical and 3 other vibrios. RESULTS: V. cholerae O139 contained the genomic sequences of ctx A2-B as well as V. cholerae O1. V. cholerae and others vibrios were divided into 4 groups by fingerprint patterns of RAPD, that is (1) V. cholerae O139 and V. cholerae O1 El Tor; (2) V. cholerae O1 classical; (3) V. paraheamolyticus and V. vulnificus and (4) V. flluvialis. V. cholerae O139 DNA fingerprint of RAPD was consistent with the El Tor biotype: average linkage cluster distance was 0, and slightly different from the classical biotype, with a distance of 2.07. It was much more different from vibrio paraheamolyticus and others, with a distance of 6.76 - 8.54. CONCLUSION: V. cholerae and other vibrios are polymorphic in inherent characteristics. The inherent characteristics of V. cholerae O139 are the same as El Tor biotype. O139 may have evolved from the El Tor biotype. The inherent characteristics of vibrio paraheamolyticus are the same as vibrio vulnificus.

DNA, Bacterial↗
PubMed2003

Vibrio fortis sp. nov. and Vibrio hepatarius sp. nov., isolated from aquatic animals and the marine environment.

In this study, the taxonomic positions of 19 Vibrio isolates disclosed in a previous study were evaluated. Phylogenetic analysis based on 16S rDNA sequences partitioned these isolates into groups that were closely related (98.8-99.1 % similarity) to Vibrio pelagius and Vibrio xuii, respectively. DNA-DNA hybridization experiments further showed that these groups had <70 % similarity to other Vibrio species. Two novel Vibrio species are proposed to accommodate these groups: Vibrio fortis sp. nov. (type strain, LMG 21557(T)=CAIM 629(T)) and Vibrio hepatarius sp. nov. (type strain, LMG 20362(T)=CAIM 693(T)). The DNA G+C content of both novel species is 45.6 mol%. Useful phenotypic features for discriminating V. fortis and V. hepatarius from other Vibrio species include production of indole and acetoin, utilization of cellobiose, fermentation of amygdalin, melibiose and mannitol, beta-galactosidase and tryptophan deaminase activities and fatty acid composition.

Animals↗
PubMed1982

Distribution and ecology of Vibrio vulnificus and other lactose-fermenting marine vibrios in coastal waters of the southeastern United States.

Water, sediment, plankton, and animal samples from five coastal sites from North Carolina to Georgia were sampled for their lactose-fermenting vibrio populations. Over 20% of all vibrios tested were sucrose negative and o-nitrophenyl-beta-D-galactopyranoside (ONPG) positive, suggesting identification as the human pathogen Vibrio vulnificus. These vibrios were isolated from all sample sites and sources (water, sediment, plankton, and animals). Correlations with several of 19 environmental parameters monitored at each site were found for total vibrios. The presence of ONPG-positive, sucrose-negative vibrios was correlated with hydrocarbon levels in the water and, in the case of plankton samples, with salinity. A total of 279 sucrose-negative, ONPG-positive isolates were subjected to numerical taxonomic analysis, which resulted in three major clusters. Cluster I corresponded to and included 11 reference strains of V. vulnificus. Cluster II contained the largest number (133) of isolates, of which the great majority were bioluminescent. Although having a resemblance to V. harveyi, the isolates were ONPG positive and many were H2S positive. Cluster III consisted of strains similar to the group F vibrios (V. fluvialis). Of all of the isolates, 55% were luminescent, of which over 20% were lethal when injected into mice. Problems involved in detecting lactose fermentation among marine vibrios and the potential pathogenicity of these organisms are discussed.

Animals↗
PubMed1982

[Quantitative evaluation of Vibrio cholerae colonization and lymphocyte count in small intestine villi of rabbits immunized with vibrio and cholera toxin].

The influence of parenteral immunization with live virulent vibrios and lipopolysaccharide-containing choleragen toxoid and oral immunization with liver avirulent vibrios on the interaction of V. eltor and the mucous membrane was studied on the ligated loops of the small intestine in 164 rabbits. Protection from enterotoxigenicity (the accumulation of fluid) was compared with the suppressive effect on the adhesion and colonization of vibrios. The coefficient of immunization efficacy was determined microbiologically, as the ratio of the adhesion index (i. e. the percentage of the number of vibrios in the media inoculated with the homogenized matter of the washed intestinal wall from the sum of this number and the amount of vibrios in the contents) in the control rabbits to the adhesion index in the immunized ones, and histologically, as the ratio of the average number of vibrios attached to the epithelial number of vibrios attached to the epithelial cells of one intestinal villus in the control rabbits to the corresponding number in the immunized animals. Protection from the enterotoxigenicity of vibrios resulted from the suppression of their adhesion and colonization, lipopolysaccharide being, probably, the factor of adhesiveness. Effective protection was accompanied by an increase in the number of lymphocytes (especially "granular" lymphocytes with lysosome-like granules) between intestinal villi and the number of lymphoid and plasmic cells in the lamina propria of the villi, which indicates the role of lymphocytes in cholera-induced immunity.

Adhesiveness↗
PubMed1997

High Prevalence of Vibrio cholerae Non-O1 Carrying Heat-Stable-Enterotoxin-Encoding Genes among Vibrio Isolates from a Temperate-Climate River Basin of Central Italy.

Vibrio spp. of clinical interest from the Arno River basin (Tuscany, Italy) were investigated in this study. Vibrios were isolated from 70% of water samples. Vibrio cholerae non-O1 was the most prevalent species (82% of isolates), followed by Vibrio mimicus (10%) and Vibrio metschnikovii (8%). Recovery of vibrios was correlated with temperature, pH, and various indicators of municipal pollution. None of the 150 Vibrio isolates carried ctx-related genomic sequences, whereas 18 (14.6%) of the 123 V. cholerae non-O1 isolates and 1 (6.7%) of the 15 V. mimicus isolates carried sto alleles. These findings indicate that considerable circulation of sto-positive vibrios may occur in temperate-climate freshwater environments.

Journal Article↗
PubMed2000

Distribution of Vibrio cholerae virulence genes among different Vibrio species isolated in Sardinia, Italy.

The members of the genus Vibrio include harmless aquatic strains as well as strains capable of causing epidemics of cholera. Diarrhoea caused by Vibrio cholerae is attributed to cholerae enterotoxin (CT) codified by the ctx operon and regulated by a number of virulence genes such as toxT, toxR and toxS. Fifty-two Vibrio strains were isolated from different aquatic environments in and around Sardinia and searched by PCR for the presence of ctxA, zot, ace, toxR, toxS, toxT, tcpA and vpi virulence genes in the genomes of the isolates. The toxR operon was found in 27 Vibrio alginolyticus strains out of 42 analysed, in three out of four V. cholerae non-O1 strains and in three Vibrio parahaemolyticus isolates. A positive amplification for the virulence pathogenic island (vpi) was produced by five V. alginolyticus strains. Finally, the ace expected amplification fragment was found in two V. alginolyticus isolates whereas the amplification with zot primers produced the expected fragment in one V. alginolyticus isolate. Differentiation of these strains with a PCR fingerprinting technique revealed no association between the presence of virulence genes and a particular fingerprinting pattern. Although most Vibrio species are considered non-pathogenic or only potentially harmful to humans, the finding of V. cholerae virulence genes in other members of the genus Vibrio, and the recent reports of the creation and evolution of pandemic strains of V. cholerae, may give a new perspective to the significance of these results.

DNA Fingerprinting↗
PubMed1978

Ecological relationship between Vibrio parahaemolyticus and agar-digesting vibrios as evidenced by bacteriophage susceptibility patterns.

Twenty bacteriophages active against Vibrio parahaemolyticus and agar-digesting vibrios, isolated from oysters (Crassostrea gigas) and Dungeness crab (Cancer magister) and by induction of a lysogenic agar digester, were tested as to their host range. These phages were specific for V. parahaemolyticus and various agar-digesting vibrios, and interspecies lysis occurred only between these two groups. V. alginolyticus, V. anguillarum and related species, V. cholerae, and a group of marine psychrophilic and psychrotrophic vibrios were not affected. No correlation was observed between the O and K serotypes of V. parahaemolyticus strains and bacteriophage susceptibility patterns, and 7 of 28 strains of V. parahaemolyticus were not lysed by any of the phages. Only two of the phage isolates were capable of lysing all susceptible V. parahaemolyticus strains. No correlation was observed between the inter-and intraspecies genetic relatedness (DNA homologies) of V. parahaemolyticus and agar-digesting vibrios and susceptibility patterns to different bacteriophages. Some of the phages were capable of plaque formation on V. parahaemolyticus as well as on some strains of agar-digesting vibrios that were separated by 70 to 80% differences in their DNA homologies. The possible ecological significance of these vibrio bacteriophages, particularly those having a wide host range, is discussed.

Agar↗
PubMed1999

Occurrence, diversity, and pathogenicity of halophilic Vibrio spp. and non-O1 Vibrio cholerae from estuarine waters along the Italian Adriatic coast.

The occurrence, diversity, and pathogenicity of Vibrio spp. were investigated in two estuaries along the Italian Adriatic coast. Vibrio alginolyticus was the predominant species, followed by Vibrio parahaemolyticus, non-O1 Vibrio cholerae, and Vibrio vulnificus. By using a biochemical fingerprinting method, all isolates were grouped into nine phenotypes with similarity levels of 75 to 97.5%. The production of toxins capable of causing cytoskeleton-dependent changes was detected in a large number of Vibrio strains. These findings indicate a significant presence of potentially pathogenic Vibrio strains along the Adriatic coast.

Animals↗
PubMed2002

Effects of depuration of molluscs experimentally contaminated with Escherichia coli, Vibrio cholerae 01 and Vibrio parahaemolyticus.

AIMS: The aim of the present study was to investigate the behaviour of two pathogenic vibrios (Vibrio cholerae O1 and Vibrio parahaemolyticus) during depuration and to compare it with that of Escherichia coli, used as an indicator of suitability for consumption. METHODS AND RESULTS: Samples of Mytilus galloprovincialis were experimentally contaminated with E. coli, V. cholerae O1 and V. parahaemolyticus, depurated in a pilot plant using ozone and analysed at selected intervals. Numbers of E. coli and vibrios were estimated using an MPN method. The presence of vibrios was confirmed by the use of PCR. The target genes used were ctx for V. cholerae O1 and the restriction fragment pR72H for V. parahaemolyticus. There was a substantially smaller reduction in the numbers of both vibrios (approximately 1 log) during the depuration process than of E. coli (approximately 3 log). CONCLUSIONS: The results confirm the inadequacy of E. coli as an indicator that molluscs have been cleansed of other microbiological agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The study confirms the risk associated with the consumption of mussels and the need to correctly conserve and cook them prior to consumption.

Animals↗
PubMed1985

Incidence of Vibrio cholerae and related vibrios in a coastal lagoon and seawater influenced by lake discharges along an annual cycle.

Most probable numbers of Vibrio cholerae and related vibrios were determined in Albufera Lake, Valencia, Spain, and in coastal waters under the influence of the lake discharges over the course of an annual cycle. The influence of temperature, kind of water, and characteristics of the different sampling sites on the numbers of vibrios recovered was evaluated. Maximum recovery of vibrios reached 10(3)/ml in both types of waters analyzed. V. cholerae numbers reached 10(3)/ml in the lake and 10(2) in one of the coastal sites. Frequently during the warm season, all vibrios isolated were identified as V. cholerae. Occasionally, no V. cholerae was recovered. The recovery of vibrios was significantly influenced by the temperature of the water and the type of water analyzed. Most of the V. cholerae isolates were included in Heiberg groups I and II, and nearly 50% of the strains used chitin as sole carbon source. Indole was not produced by 100% of the strains. All strains tested were non-O1 serovars.

Chitin↗
PubMed1987

New selective and differential medium for Vibrio cholerae and Vibrio vulnificus.

Thiosulfate-citrate-bile salts-sucrose agar has been routinely used for the isolation of pathogenic vibrios, although its selectivity for Vibrio cholerae and Vibrio vulnificus is inadequate. Therefore, a new plating medium, cellobiose-polymyxin B-colistin agar, was developed for the isolation of these two species. Cellobiose-polymyxin B-colistin agar demonstrated a significant advantage over other media designed for the isolation or differentiation of vibrios: of both the 136 strains representing 19 Vibrio species and the marine isolates of the genera Pseudomonas, Flavobacterium, and Photobacterium, only V. vulnificus and V. cholerae were able to grow. Furthermore, the fermentation of cellobiose by V. vulnificus allowed for the easy differentiation of these two species. This medium offers significant potential as a selective and differential medium for these two pathogenic vibrios.

Culture Media↗
PubMed2000

The marine pathogen Vibrio vulnificus encodes a putative homologue of the Vibrio harveyi regulatory gene, luxR: a genetic and phylogenetic comparison.

Vibrio vulnificus is an opportunistic pathogen that exhibits numerous virulence factors, including the secretion of a zinc metalloprotease and the production of a capsule. We have cloned and sequenced a gene from V. vulnificus that is a homologue of the positive transcriptional regulator, luxR, of the lux operon in Vibrio harveyi. This gene encodes a putative, single complete open reading frame designated smcR, which shares greater than 75% nucleotide identity with luxR of V. harveyi. The deduced amino acid sequence of the putative SmcR protein is more than 90% identical and 95% similar to that of LuxR of V. harveyi, suggesting that V. vulnificus possesses a member of the family of signal-response genes recently described in Vibrio cholerae and in Vibrio parahaemolyticus. Our data also demonstrate that, in addition to V. vulnificus, all six Vibrio spp. tested contained genes that hybridized with the luxR probe. We also present evidence that this regulatory protein was inherited from a common ancestor, and that the gene is ancient and widespread in marine Vibrio spp.

Amino Acid Sequence↗
PubMed1989

Vibrio parahaemolyticus and other halophilic vibrios associated with seafood in Hong Kong.

The summer prevalence of Vibrio parahaemolyticus and other halophilic vibrios in seafood from Hong Kong markets was investigated. Halophilic vibrios were isolated from all seven types of seafood examined, and comprised 9.1%, 8% and 6.1% of contaminating aerobic heterotrophic bacteria from mussels, clams and oysters respectively. Sucrose-positive vibrios were more common than sucrose-negative varieties. Vibrio alginolyticus was the most frequently isolated species, followed by V. parahaemolyticus, V. harveyi, V. fluvialis, V. vulnificus, V. pelagius, V. campbellii, V. spendidus and V. marinus. Mussels contained the highest concentration of V. parahaemolyticus (4.6 x 10(4)/g); oysters and clams contained 3.4 x 10(4)/g and 6.5 x 10(3)/g respectively. The ubiquity and relatively high concentrations of V. parahaemolyticus and other pathogenic vibrios in shellfish is a potential public health hazard in Hong Kong and other subtropical Asian countries.

Animals↗
PubMed1983

Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment.

During the summer of 1981, 3,887 sucrose-negative vibrios were isolated from seawater, sediment, plankton, and animal samples taken from 80 sites from Miami, Fla., to Portland, Maine. Of these, 4.2% were able to ferment lactose. The lactose-positive strains isolated from the various samples correlated positively with pH and turbidity of the water, vibrios in the sediment and oysters, and total bacterial counts in oysters. Negative correlations were obtained for water salinity. Numerical taxonomy was performed on 95 of the lactose-fermenting environmental isolates and 23 reference strains. Five clusters resulted, with the major cluster containing 33 of the environmental isolates and all of the Vibrio vulnificus reference strains. The 33 isolates, which produced an acid reaction in lactose broth within hours of initial inoculation, represented 20% of all lactose-fermenting vibrios studied. These isolates were nearly identical phenotypically to clinical strains of V. vulnificus studied by the Centers for Disease Control, Atlanta, Ga., and by our laboratory, and their identification was confirmed by DNA-DNA hybridization studies. V. vulnificus was isolated from all sample types and from Miami to Cape Cod, Mass., and comparison of the environmental parameters of the eight subsites yielding this species with those of all 80 subsites revealed no significant differences. The majority of the isolates were obtained from animals, with clams providing most (84%) of these. On injection into mice, 82% of the V. vulnificus isolates resulted in death. Members of the remaining four clusters contained strains which differed from V. vulnificus in such phenotypic traits as luminescence and in urease or H(2)S production. None of the other reference cultures, including nine other Vibrio species, were contained in the remaining clusters, and these isolates could not be identified. Most of these were also lethal for mice. Phenotypic differences, potential pathogenicity, and geographic distribution of the five clusters were examined. It is concluded that V. vulnificus is a ubiquitous organism, both geographically and in a variety of environmental sources, although it occurs in relatively low numbers. The public health significance of this organism and of the other unidentified lactose-fermenting Vibrio species is discussed.

Fermentation↗
PubMed1993

Extraintestinal infections caused by Vibrio parahaemolyticus and Vibrio alginolyticus in a Danish county, 1987-1992.

Over a 6-year period we detected 30 clinical infections caused by halophilic vibrios in a restricted geographical area. Vibrio parahaemolyticus infections were found in 13 patients (3 with wound infections, 10 with ear infections), and Vibrio alginolyticus infections in 17 patients, all of whom had ear infections. From 1987 to 1990, infections caused by marine vibrios were found in at most 4 cases annually, in 1992 in 6 instances, whereas in a 5-month period in 1991 we experienced 15 cases of extra-intestinal infections. 15 of the infections found in 1991 and 1992 were ear infections, 10 of which occurred shortly after exposure to Danish coastal seawater. The disease was mild in most of the patients and all recovered. Most of the patients showed predisposing conditions such as chronic otitis media, perforation of the tympanic membrane or ulcus cruris. The organisms were isolated in minor numbers from coastal water samples from 5 of 12 bathing areas around Funen at the end of July, but not at the end of November. This study indicates that the halophilic marine vibrios may be pathogenic in Scandinavian areas in persons exposed to seawater.

Adolescent↗
PubMed1991

[Surveys on the contamination of marine fish with non-O1 Vibrio cholerae and Vibrio mimicus and food poisoning cases by these organisms].

The present paper describes the relationship between the contamination with non-O1 Vibrio cholerae and Vibrio mimicus of marine fish, with special reference to the seasonal variation and the concentration of contamination, and the actual cases of domestic food poisoning by these organisms. A 10 year survey revealed that non-O1 Vibrio cholerae (non-O1 V. cholerae) strains were frequently isolated from fish during the summer season with some variations from one year to another, and isolates from fish showed similar biological properties to those of isolates from diarrhea cases of over-sea travellers. Experimentally enteropathogenic strains were included among these isolates. Vibrio mimicus (V. mimicus) strains were also isolated from fish, the frequency being not so high as in the case of non-O1 V. cholerae Strains of serovar O-41 which was most predominant among strains from diarrhea cases were also detected among the isolates from fish. The viable cell counts, however, were very small with regard to both non-O1 V. cholerae and V. mimicus From these observations, factors causing food poisoning by non-O1 V. cholerae or V. mimicus seemed to be essentially similar to those by Vibrio parahaemolyticus (V. parahaemolyticus); that is, the food poisoning by non-O1 V. cholerae or V. mimicus is apt to occur in the summer season and is caused by the consumption of raw fish, although the frequency might be significantly low in comparison to that of V. parahaemolyticus. The actual cases of the domestic food poisoning by non-O1 V. cholerae or V. mimicus were retrospectively surveyed by the literature.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1999

Randomly amplified polymorphic DNA analysis of clinical and environmental isolates of Vibrio vulnificus and other vibrio species.

Vibrio vulnificus is an estuarine bacterium that is capable of causing a rapidly fatal infection in humans. A randomly amplified polymorphic DNA (RAPD) PCR protocol was developed for use in detecting V. vulnificus, as well as other members of the genus Vibrio. The resulting RAPD profiles were analyzed by using RFLPScan software. This RAPD method clearly differentiated between members of the genus Vibrio and between isolates of V. vulnificus. Each V. vulnificus strain produced a unique band pattern, indicating that the members of this species are genetically quite heterogeneous. All of the vibrios were found to have amplification products whose sizes were within four common molecular weight ranges, while the V. vulnificus strains had an additional two molecular weight range bands in common. All of the V. vulnificus strains isolated from clinical specimens produced an additional band that was only occasionally found in environmental strains; this suggests that, as is the case with the Kanagawa hemolysin of Vibrio parahaemolyticus, the presence of this band may be correlated with the ability of a strain to produce an infection in humans. In addition, band pattern differences were observed between encapsulated and nonencapsulated isogenic morphotypes of the same strain of V. vulnificus.

Bacterial Typing Techniques↗
PubMed1976

The isolation of Vibrio parahaemolyticus and related vibrios from moribund aquarium lobsters.

Vibrios were isolated in pure culture from the hemolymph of 7 out of 28 dead or dying aquarium lobsters which had been acclimated to 20-22 degrees C. One isolate was identified as Vibrio parahaemolyticus, one as a related marine Vibrio (probably V. marinus), and five as Vibrio alginolyticus. No isolates of halophilic Vibrio species were made from healthy lobsters using thiosulfate citrate bile salts sucrose agar (TCBS).

Animals↗