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Letters in Applied Microbiology Sep 2010To develop an effective multiplex PCR for simultaneous and rapid detection of Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus, the three most important...
AIMS
To develop an effective multiplex PCR for simultaneous and rapid detection of Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus, the three most important Vibrio species that can cause devastating health hazards among human.
METHODS AND RESULTS
Species-specific PCR primers were designed based on toxR gene for V. cholerae and V. parahaemolyticus, and vvhA gene for V. vulnificus. The multiplex PCR was validated with 488 Vibrio strains including 322 V. cholerae, 12 V. vulnificus, and 82 V. parahaemolyticus, 20 other Vibrio species and 17 other bacterial species associated with human diseases. It could detect the three target bacteria without any ambiguity even among closely related species. It showed good efficiency in detection of co-existing target species in the same sample. The detection limit of all the target species was ten cells per PCR tube.
CONCLUSIONS
Specificity and sensitivity of the multiplex PCR is 100% each and sufficient for simultaneous detection of these potentially pathogenic Vibrio species in clinical and environmental samples.
SIGNIFICANCE AND IMPACT OF THE STUDY
This simple, rapid and cost-effective method can be applicable in a prediction system to prevent disease outbreak by these Vibrio species and can be considered as an effective tool for both epidemiologist and ecologist.
Topics: Bacterial Proteins; Bacteriological Techniques; DNA-Binding Proteins; Humans; Polymerase Chain Reaction; Sensitivity and Specificity; Transcription Factors; Vibrio Infections; Vibrio cholerae; Vibrio parahaemolyticus; Vibrio vulnificus
PubMed: 20666989
DOI: 10.1111/j.1472-765X.2010.02895.x -
Journal of Bacteriology Dec 2012Vibrio vulnificus, which is, like Vibrio cholerae and Vibrio parahaemolyticus, a pathogen of humankind, is a Gram-negative, curved, motile, and rod-shaped bacterium...
Vibrio vulnificus, which is, like Vibrio cholerae and Vibrio parahaemolyticus, a pathogen of humankind, is a Gram-negative, curved, motile, and rod-shaped bacterium [corrected] Here, we present the draft genome sequence of the type strain, ATCC 27562, which was the first isolated Vibrio vulnificus strain.
Topics: Base Sequence; DNA, Bacterial; Genome, Bacterial; Molecular Sequence Data; RNA, Bacterial; Sequence Analysis, DNA; Sodium Chloride; Vibrio vulnificus
PubMed: 23209214
DOI: 10.1128/JB.01890-12 -
Environmental Microbiology Aug 2019Vibrio vulnificus is a siderophilic pathogen spreading due to global warming. The zoonotic strains constitute a clonal-complex related to fish farms that are distributed...
Vibrio vulnificus is a siderophilic pathogen spreading due to global warming. The zoonotic strains constitute a clonal-complex related to fish farms that are distributed worldwide. In this study, we applied a transcriptomic and single gene approach and discover that the zoonotic strains bypassed the iron requirement of the species thanks to the acquisition of two iron-regulated outer membrane proteins (IROMPs) involved in resistance to fish innate immunity. Both proteins have been acquired by horizontal gene transfer and are contributing to the successful spreading of this clonal-complex. We have also discovered that the zoonotic strains express a virulent phenotype in the blood of its main susceptible hosts (iron-overloaded humans and healthy eels) by combining a host-specific protective envelope with the common expression of two toxins (VvhA and RtxA1), one of which (RtxA1) is directly involved in sepsis. Finally, we found that both IROMPs are also present in other fish pathogenic species and have recently been transmitted to the phylogenetic lineage involved in human primary sepsis after raw seafood ingestion. Together our results highlight the potential hazard that the aquaculture industry poses to public health, which is of particular relevance in the context of a warming world.
Topics: Acclimatization; Animals; Fish Diseases; Fishes; Gene Transfer, Horizontal; Humans; Immunity, Innate; Iron; Phylogeny; Sepsis; Vibrio Infections; Vibrio vulnificus; Zoonoses
PubMed: 31206984
DOI: 10.1111/1462-2920.14714 -
Letters in Applied Microbiology Jan 2014This study evaluated the efficacy of grape seed extract (GE), citric acid (CA) and lactic acid (LA) on the inactivation of Vibrio vulnificus and inherent microflora in...
UNLABELLED
This study evaluated the efficacy of grape seed extract (GE), citric acid (CA) and lactic acid (LA) on the inactivation of Vibrio vulnificus and inherent microflora in fresh shucked oysters. The minimum inhibitory concentration (MIC) of GE, CA or LA against V. vulnificus was determined. Furthermore, the shucked oysters were artificially inoculated with V. vulnificus. The inoculated shucked oysters (25 g) were then dipped in 250 ml GE, CA or LA solutions for 10 min. The population of V. vulnificus in shucked oysters was determined. The effects of the treatments with GE, CA or LA solutions on the inherent microbiota in fresh shucked oysters during storage at 5°C for 20 days were also studied. The MICs of GE, CA or LA against V. vulnificus were 10.0, 5.0 or 1.0 mg ml(-1), respectively. The concentrations of 500, 300 or 150 mg ml(-1) GE, CA or LA solutions were needed to reduce the population of V. vulnificus to below the detection level (1.0 log g(-1)). Treatment with 500, 300, 150 mg ml(-1) GE, CA or LA significantly reduced the initial inherent microbiota in fresh shucked oysters, and inherent levels were significantly (P < 0.05) lower than the control sample throughout refrigerated storage for 20 days.
SIGNIFICANCE AND IMPACT OF THE STUDY
Oysters filter large volume of seawater during their feeding activities that concentrate bacteria such as Vibrio vulnificus in their body. The presence of V. vulnificus in oysters has a serious impact on public health and international trade. There is increasing concern over the use of chemical preservatives. Furthermore, the food industry is looking for new natural preservation methods. This study indicated that lactic acid and citric acid wash solutions could offer an inexpensive, natural and strong approach to control V. vulnificus and spoilage bacteria in fresh shucked for the oyster industry.
Topics: Animals; Anti-Bacterial Agents; Bacterial Load; Citric Acid; Grape Seed Extract; Lactic Acid; Microbial Sensitivity Tests; Ostreidae; Seawater; Shellfish; Temperature; Vibrio vulnificus
PubMed: 24001001
DOI: 10.1111/lam.12152 -
Microbial Pathogenesis Oct 2017The re-emerging field of phage therapy is the potential biocontrol agents for the transfer of virulence factor and to kill their bacterial hosts. In this study, the...
The re-emerging field of phage therapy is the potential biocontrol agents for the transfer of virulence factor and to kill their bacterial hosts. In this study, the lytic Vibrio vulnificus phages were studied to provide a better understanding of phage-host interactions and development of phage therapy. Four new V. vulnificus phages were detected from shrimp aquaculture system, named VV1, VV2, VV3 and VV4. All lytic V. vulnificus phages are the Tectiviruses of the family Tectiviridae with typical double layered elongated icosahedral head and tailless morphology. Lytic V. vulnificus phages which infect other Vibrio isolates were further characterized by long term storage, enzymes treatment, organic solvents treatment, detergents treatment, pH stability, temperature stability, agar bioassay method and one-step growth experiment. The effects of chloroform, acetone, ethyl alcohol, methyl alcohol, ribonuclease (RNase), trypsin, protease, Triton-X100 treatments were not affected the growth of VV1, VV2, VV3 and VV4 phages. These phages (VV1-VV4) were inactivated completely with temperature (over 60 °C), pH (<3 or >12), lysozyme and sodium dodecyl sulfate (SDS) treatment. One-step growth experiments indicated that the latent period was at 3 h and burst size was at 37 °C. Agar bioassay method indicated that the percentage of inhibition was 75% (VV1) and 70% (VV2, VV3 & VV4), respectively. SDS-PAGE analysis of all V. vulnificus phages had similar protein patterns with molecular weight masses of 260, 249, 204, 148, 63, 59, 22 and 15 kDa. Hence, it could be concluded that V. vulnificus phage had a broad lytic spectrum and potential biocontrol of luminous Vibriosis in the shrimp aquaculture system.
Topics: Animals; Aquaculture; Bacteriophages; Crustacea; Seafood; Vibrio vulnificus
PubMed: 28917670
DOI: 10.1016/j.micpath.2017.09.024 -
Infection and Immunity Sep 2007Vibrio vulnificus is an estuarine bacterium that can cause primary septicemia as well as serious wound infections. Generally, clinical isolates have a high lethal effect...
Vibrio vulnificus is an estuarine bacterium that can cause primary septicemia as well as serious wound infections. Generally, clinical isolates have a high lethal effect compared with environmental isolates. However, little is known about the mechanisms by which V. vulnificus causes disease. In this study, we compared the pathogenicity of a clinical isolate, strain M2799, with that of an environmental isolate, strain JCM3731. The clinical isolate showed 100 times higher lethality in mice than the environmental isolate. In strain M2799-inoculated mice, the number of macrophages decreased significantly, whereas there was no appreciable change in the number of macrophages in strain JCM3731-inoculated mice. The clinical isolate showed high cytotoxic activity, especially to macrophages, compared with the environmental isolate in vitro. The growth of the clinical isolate was almost completely inhibited in the presence of macrophages. Moreover, the survival rate of the clinical isolate-inoculated mice increased by recruitment of macrophages. These results indicate that V. vulnificus infection progresses by damage to macrophages during the early phase of infection.
Topics: Animals; Disease Models, Animal; Female; Macrophages; Mice; Mice, Inbred BALB C; Vibrio Infections; Vibrio vulnificus
PubMed: 17591793
DOI: 10.1128/IAI.00481-07 -
Current Microbiology Jun 2014Gram-negative bacteria use tripartite pumps to transport antibacterial drugs and other toxic compounds across the inner and outer membranes, which are separated by the...
Gram-negative bacteria use tripartite pumps to transport antibacterial drugs and other toxic compounds across the inner and outer membranes, which are separated by the periplasmic space. The TolC protein is an outer membrane factor that participates in the formation of tripartite efflux pumps. The genome of Vibrio vulnificus encodes two E. coli TolC homologs, TolCV1 and TolCV2. Here, we show that both TolCV1 and TolCV2 are involved in the efflux of antimicrobial agents. Deletion of tolCV1 resulted in increased susceptibility of V. vulnificus to chemical detergents, DNA intercalating agents, and antibiotics including erythromycin, novobiocin, and tetracycline, whereas deletion of tolCV2 rendered V. vulnificus more susceptible to the above mentioned antibiotics only. We also observed that tolCV1 deletion resulted in reduced motility of V. vulnificus. Our results indicate active roles for TolCV1 and TolCV2 in the physiology of V. vulnificus.
Topics: Anti-Infective Agents; Bacterial Outer Membrane Proteins; Gene Deletion; Membrane Transport Proteins; Microbial Sensitivity Tests; Vibrio vulnificus
PubMed: 24515351
DOI: 10.1007/s00284-014-0537-4 -
Yakugaku Zasshi : Journal of the... Jul 2005Bacteria of the genus Vibrio are normal habitants of the aquatic environment and play roles for biocontrole of aquatic ecosystem, but some species are believed to be... (Review)
Review
Bacteria of the genus Vibrio are normal habitants of the aquatic environment and play roles for biocontrole of aquatic ecosystem, but some species are believed to be human pathogens. These species can be classified into two groups according to the types of diseases they cause: the gastrointestinal infections and the extraintestinal infections. The pathogenic species produce various pathogenic factors including enterotoxin, hemolysin, cytotoxin, protease, siderophore, adhesive factor, and hemagglutinin. We studied various pathogenic factors of vibrios with special emphasis on protease and hemolysin of V. vulnificus. V. vulnificus is now recognized as being among the most rapidly fatal of human pathogens, although the infection is appeared in patients having underlying disease(s) such as liver dysfunction, alcoholic cirrhosis or haemochromatosis. V. vulnificus protease (VVP) is thought to be a major toxic factor causing skin damage in the patients having septicemia. VVP is a metalloprotease and degrades a number of biologically important proteins including elastin, fibrinogen, and plasma proteinase inhibitors of complement components. VVP causes skin damages through activation of the Factor XII-plasma kallikrein-kinin cascade and/or exocytotic histamine release from mast cells, and a haemorrhagic lesion through digestion of the vascular basement membrane. Thus, the protease is the most probable candidate for tissue damage and bacterial invasion during an infection. Pathogenic roles and functional mechanism of other factors including hemolysins of V. vulnificus and V. mimicus are also shown in this review article.
Topics: Animals; Bacterial Proteins; Endopeptidases; Gastroenteritis; Hemolysin Proteins; Humans; Immunocompromised Host; Metalloproteases; Siderophores; Skin Diseases, Infectious; Vibrio Infections; Vibrio mimicus; Vibrio vulnificus; Virulence Factors
PubMed: 15997210
DOI: 10.1248/yakushi.125.531 -
Antonie Van Leeuwenhoek Nov 2018Vibrio vulnificus, a Gram-negative halophilic bacterium, is an opportunistic human pathogen that is responsible for the majority of seafood-associated deaths worldwide....
Vibrio vulnificus, a Gram-negative halophilic bacterium, is an opportunistic human pathogen that is responsible for the majority of seafood-associated deaths worldwide. Lipoproteins are important components of the bacterial cell envelope and have been shown to be involved in a wide variety of cellular processes. Little is known about the localisation or transport mechanism of lipoproteins in V. vulnificus. To assess the localisation of lipoproteins in V. vulnificus, we tested two established techniques for the rapid separation of membrane-associated proteins: detergent extraction with Sarkosyl and outer membrane vesicles (OMVs) preparation. The results showed that Sarkosyl extraction was not useful for the separation of lipoproteins from the different membranes of V. vulnificus. On the other hand, we confirmed that OMVs produced by V. vulnificus contained lipoproteins from the outer but not the inner membrane. Analysis of the OMVs components confirmed the localisation of several well-known lipoproteins to membranes that were different from expected, based on their predicted functions. Using this technique, we found that Asp at position +2 of mature lipoproteins can function as an inner membrane retention signal in V. vulnificus. Interestingly, the Escherichia coli "+2 rule" does not apply to the V. vulnificus lipoprotein IlpA (G2D) mutant, as a Ser to Asp mutation at position +2 of IlpA did not affect its outer membrane localisation. Furthermore, an IlpA tether-mRFP chimeric lipoprotein and its G2D mutant also behaved like IlpA. Together, these results suggest that the sorting rule of lipoproteins in V. vulnificus might be different from that in E. coli.
Topics: Bacterial Outer Membrane Proteins; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Lipoproteins; Microscopy, Electron, Transmission; Mutation; Sarcosine; Vibrio vulnificus
PubMed: 29721710
DOI: 10.1007/s10482-018-1092-y -
FEMS Immunology and Medical Microbiology Dec 2011Curcumin, a natural polyphenolic flavonoid extracted from the rhizome of Curcuma longa L., has many beneficial biological activities. However, there are relatively few...
Curcumin, a natural polyphenolic flavonoid extracted from the rhizome of Curcuma longa L., has many beneficial biological activities. However, there are relatively few reports of the effects of curcumin on pathogen infections. This study examined the effect of curcumin on a Vibrio vulnificus infection. The cytotoxicity of V. vulnificus to HeLa cells was significantly inhibited by curcumin (at 10 or 30 μM). To further examine the inhibitory mechanism of curcumin against V. vulnificus-mediated cytotoxicity, the level of bacterial growth, bacterial motility, cell adhesion, RTX toxin expression and host cell reactions were evaluated. Curcumin inhibited V. vulnificus growth in HI broth. Curcumin inhibited both bacterial adhesion and RTX toxin binding to the host cells, which can be considered the major protective mechanisms for the decrease in V. vulnificus cytotoxicity. Curcumin also inhibited host cell rounding and actin aggregation, which are the early features of cell death caused by V. vulnificus. In addition, curcumin decreased the V. vulnificus-induced NF-κB translocation in HeLa cells. Finally, curcumin protected mice from V. vulnificus-induced septicemia. In conclusion, curcumin may be an alternative antimicrobial agent against fatal bacterial infections.
Topics: Animals; Anti-Bacterial Agents; Bacterial Toxins; Cell Adhesion; Curcumin; Disease Models, Animal; Epithelial Cells; HeLa Cells; Humans; Locomotion; Mice; NF-kappa B; Vibrio Infections; Vibrio vulnificus
PubMed: 22092562
DOI: 10.1111/j.1574-695X.2011.00855.x