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1

Effect of organic acids on shrimp pathogen, Vibrio harveyi.  

PubMed

Shrimp farming accounts for more than 40% of the world shrimp production. Luminous vibriosis is a shrimp disease that causes major economic losses in the shrimp industry as a result of massive shrimp kills due to infection. Some farms in the South Asia use antibiotics to control Vibrio harveyi, a responsible pathogen for luminous vibriosis. However, the antibiotic-resistant strain was found recently in many shrimp farms, which makes it necessary to develop alternative pathogen control methods. Short-chain fatty acids are metabolic products of organisms, and they have been used as food preservatives for a long time. Organic acids are also commonly added in feeds in animal husbandry, but not in aquaculture. In this study, growth inhibitory effects of short-chain fatty acids, namely formic acid, acetic acid, propionic acid, and butyric acid, on V. harveyi were investigated. Among four acids, formic acid showed the strongest inhibitory effect followed by acetic acid, propionic acid, and butyric acid. The minimum inhibitory concentration (MIC) of 0.035% formic acid suppressed growth of V. harveyi. The major inhibitory mechanism seems to be the pH effect of organic acids. The effective concentration 50 (EC50) values at 96 h inoculation for all organic acids were determined to be 0.023, 0.041, 0.03, and 0.066% for formic, acetic, propionic, and butyric acid, respectively. The laboratory study results are encouraging to formulate shrimp feeds with organic acids to control vibrio infection in shrimp aquaculture farms. PMID:21479932

Mine, Saori; Boopathy, Raj

2011-07-01

2

Quorum Sensing-Disrupting Brominated Furanones Protect the Gnotobiotic Brine Shrimp Artemia franciscana from Pathogenic Vibrio harveyi, Vibrio campbellii, and Vibrio parahaemolyticus Isolates  

Microsoft Academic Search

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

Tom Defoirdt; Roselien Crab; Thomas K. Wood; Patrick Sorgeloos; Willy Verstraete; Peter Bossier

2006-01-01

3

Molecular identification of pathogenic and nonpathogenic strains of Vibrio harveyi using PCR and RAPD  

Microsoft Academic Search

Fifteen environmental samples of Vibrio spp. isolated from healthy and diseased shrimps were tested for pathogenicity to juvenile shrimps. Two isolates, strains Z2 and Z3, were observed to be pathogenic, causing 100% mortality of the target host compared to the control strain Vibrio harveyi ATCC 14126. Environmental and type strains were subjected to molecular characterization by restriction fragment length polymorphism

G. Hernández; J. Olmos

2004-01-01

4

Quorum Sensing-Disrupting Brominated Furanones Protect the Gnotobiotic Brine Shrimp Artemia franciscana from Pathogenic Vibrio harveyi, Vibrio campbellii, and Vibrio parahaemolyticus Isolates†  

PubMed Central

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.

Defoirdt, Tom; Crab, Roselien; Wood, Thomas K.; Sorgeloos, Patrick; Verstraete, Willy; Bossier, Peter

2006-01-01

5

Draft Genome Sequence Of The Shrimp Pathogen Vibrio harveyi CAIM 1792.  

National Technical Information Service (NTIS)

Vibrio harveyi is a Gram-negative bacterium found in tropical and temperate marine environments as a free-living organism or in association with aquatic animals. We report the first sequenced genome of a Vibrio harveyi strain, CAIM 1792, the etiologic age...

G. J. Vora I. Espinoza-Valles R. A. Edwards S. Soto-Rodr guez Z. Wang

2012-01-01

6

Antibacterial effect of medium-chain fatty acid: caprylic acid on gnotobiotic Artemia franciscana nauplii against shrimp pathogens Vibrio harveyi and V. parahaemolyticus  

Microsoft Academic Search

The present study was carried out to determine the antibacterial effect of caprylic acid in the culture system of Artemia franciscana nauplii inoculated with shrimp pathogens Vibrio harveyi and V. parahaemolyticus. To begin with, the antibacterial effect of different concentrations (1, 10 and 100 mM) of caprylic acid against V. harveyi and V. parahaemolyticus was assessed through bacterial growth study. This

G. Immanuel; M. Sivagnanavelmurugan; A. Palavesam

2011-01-01

7

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

PubMed

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. PMID:10806366

McDougald, D; Rice, S A; Kjelleberg, S

2000-05-01

8

Characterization of DegQVh, a Serine Protease and a Protective Immunogen from a Pathogenic Vibrio harveyi Strain? †  

PubMed Central

Vibrio harveyi is an important marine pathogen that can infect a number of aquaculture species. V. harveyi degQ (degQVh), the gene encoding a DegQ homologue, was cloned from T4, a pathogenic V. harveyi strain isolated from diseased fish. DegQVh was closely related to the HtrA family members identified in other Vibrio species and could complement the temperature-sensitive phenotype of an Escherichia coli strain defective in degP. Expression of degQVh in T4 was modulated by temperature, possibly through the ?E-like factor. Enzymatic analyses demonstrated that the recombinant DegQVh protein expressed in and purified from E. coli was an active serine protease whose activity required the integrity of the catalytic site and the PDZ domains. The optimal temperature and pH of the recombinant DegQVh protein were 50°C and pH 8.0. A vaccination study indicated that the purified recombinant DegQVh was a protective immunogen that could confer protection upon fish against infection by V. harveyi. In order to improve the efficiency of DegQVh as a vaccine, a genetic construct in the form of the plasmid pAQ1 was built, in which the DNA encoding the processed DegQVh protein was fused with the DNA encoding the secretion region of AgaV, an extracellular ?-agarase. The E. coli strain harboring pAQ1 could express and secrete the chimeric DegQVh protein into the culture supernatant. Vaccination of fish with viable E. coli expressing chimeric degQVh significantly (P < 0.001) enhanced the survival of fish against V. harveyi challenge, which was possibly due to the relatively prolonged exposure of the immune system to the recombinant antigen produced constitutively, albeit at a gradually decreasing level, by the carrier strain.

Zhang, Wei-wei; Sun, Kun; Cheng, Shuang; Sun, Li

2008-01-01

9

Evaluation of the vaccine potential of a cytotoxic protease and a protective immunogen from a pathogenic Vibrio harveyi strain.  

PubMed

Vibrio harveyi is an important aquaculture pathogen that can infect a number of fish species and marine invertebrates. A putative protease, Vhp1, was identified from a pathogenic V. harveyi strain isolated from diseased fish as a protein with secretion capacity. Vhp1 is 530 amino acids in length and shares high sequence identities with several extracellular serine proteases of the Vibrio species. In silico analysis identified a protease domain in Vhp1, which is preceded by a subtilisin-N domain and followed by a bacterial pre-peptidase C-terminal domain. Purified recombinant protein corresponding to the protease domain of Vhp1 exhibited apparent proteolytic activity that was relatively heat-stable and reached maximum at pH 8.0 and 50 degrees C. The activity of purified recombinant Vhp1 protease was enhanced by Ca(2+) and inhibited by Mn(2+) and ethylenedinitrilotetraacetic acid. Cytotoxicity analyses indicated that recombinant Vhp1 protease was toxic to cultured Japanese flounder cells and could cause complete cell lysis. Immunoprotective analysis using Japanese flounder as an animal model showed that purified recombinant Vhp1 in the form of a denatured and proteolytically inactive protein was an effective subunit vaccine. To improve the vaccine potential of Vhp1, an Escherichia coli strain that expresses and secrets a cytotoxically impaired Vhp1 was constructed, which, when used as a live vaccine, afforded a high level of protection upon the vaccinated fish against lethal V. harveyi challenge. Taken together, these results demonstrate that Vhp1 is a cytotoxic protease and an effective vaccine candidate against V. harveyi infection. PMID:19897068

Cheng, Shuang; Zhang, Wei-wei; Zhang, Min; Sun, Li

2010-01-22

10

Cross-species induction of luminescence in the quorum-sensing bacterium Vibrio harveyi.  

PubMed Central

Different species of bacteria were tested for production of extracellular autoinducer-like activities that could stimulate the expression of the luminescence genes in Vibrio harveyi. Several species of bacteria, including the pathogens Vibrio cholerae and Vibrio parahaemolyticus, were found to produce such activities. Possible physiological roles for the two V. harveyi detection-response systems and their joint regulation are discussed.

Bassler, B L; Greenberg, E P; Stevens, A M

1997-01-01

11

Interactions of microorganisms isolated from gilthead sea bream, Sparus aurata L., on Vibrio harveyi, a pathogen of farmed Senegalese sole, Solea senegalensis (Kaup).  

PubMed

Four bacterial isolates from farmed gilthead sea bream, Sparus aurata, included in a previous study as members of the Vibrionaceae and Pseudomonodaceae and the genus Micrococcus, have been evaluated for their adhesive ability to skin and intestinal mucus of farmed Senegalese sole, Solea senegalensis, and their antagonistic effect on Vibrio harveyi, a pathogen of sole. These isolates showed higher adhesion to sole mucus than the pathogenic strains of V. harveyi assayed. Only two of the isolates showed antagonistic activity to V. harveyi. Interactions of the four isolates with V. harveyi in respect of adhesion to skin and intestinal mucus under exclusion, competition and displacement conditions were studied. Three isolates were able to reduce the attachment to skin and intestinal sole mucus of a pathogenic strain of V. harveyi under displacement and exclusion conditions, but not under competition conditions. The in vivo probiotic potential of isolate Pdp11 was assessed by oral administration followed by challenge with the pathogenic V. harveyi strain Lg14/00. A group of 50 Senegalese sole received a commercial diet supplemented with 10(8) cfu g(-1) of lyophilized Lg14/00 for 15 days. A second group of fish received a non-supplemented commercial diet. After challenge the mortality of the fish receiving the diet supplemented with the potential probiotic isolate was significantly lower than that in the fish receiving the non-supplemented commercial diet. This study has shown that the ability to interfere with attachment of pathogens, as well as the adhesion to host surfaces, are suitable criteria for selection of candidate probiotics for use in the culture of Senegalese sole. PMID:16266326

Chabrillón, M; Rico, R M; Arijo, S; Díaz-Rosales, P; Balebona, M C; Moriñigo, M A

2005-09-01

12

Evolution of tolerance to PCBs and susceptibility to a bacterial pathogen (Vibrio harveyi) in Atlantic killifish (Fundulus heteroclitus) from New Bedford (MA, USA) harbor  

PubMed Central

A population of the non-migratory estuarine fish Fundulus heteroclitus (Atlantic killifish) resident to New Bedford (NB), Massachusetts, USA, an urban harbor highly contaminated with polychlorinated biphenyls (PCBs), demonstrates recently evolved tolerance to some aspects of PCB toxicity. PCB toxicology, ecological theory, and some precedence supported expectations of increased susceptibility to pathogens in NB killifish. However, laboratory bacterial challenges of the marine pathogen Vibrio harveyi to wild fish throughout the reproductive season and to their mature laboratory-raised progeny demonstrated comparable survival by NB and reference killifish, and improved survival by NB males. These results are inconsistent with hypothesized tradeoffs of adaptation, and suggest that evolved tolerance in NB killifish may include mechanisms that minimize the immunosuppressive effects of PCBs. Compensatory strategies of populations persisting in highly contaminated environments provide a unique perspective for understanding the long-term ecological effects of toxic chemicals.

Nacci, Diane; Huber, Marina; Champlin, Denise; Jayaraman, Saro; Cohen, Sarah; Gauger, Eric; Fong, Allison; Gomez-Chiarri, Marta

2009-01-01

13

Evolution of tolerance to PCBs and susceptibility to a bacterial pathogen (Vibrio harveyi) in Atlantic killifish (Fundulus heteroclitus) from New Bedford (MA, USA) harbor.  

PubMed

A population of the non-migratory estuarine fish Fundulus heteroclitus (Atlantic killifish) resident to New Bedford (NB), Massachusetts, USA, an urban harbor highly contaminated with polychlorinated biphenyls (PCBs), demonstrates recently evolved tolerance to some aspects of PCB toxicity. PCB toxicology, ecological theory, and some precedence supported expectations of increased susceptibility to pathogens in NB killifish. However, laboratory bacterial challenges of the marine pathogen Vibrio harveyi to wild fish throughout the reproductive season and to their mature laboratory-raised progeny demonstrated comparable survival by NB and reference killifish, and improved survival by NB males. These results are inconsistent with hypothesized trade-offs of adaptation, and suggest that evolved tolerance in NB killifish may include mechanisms that minimize the immunosuppressive effects of PCBs. Compensatory strategies of populations persisting in highly contaminated environments provide a unique perspective for understanding the long-term ecological effects of toxic chemicals. PMID:19110353

Nacci, Diane; Huber, Marina; Champlin, Denise; Jayaraman, Saro; Cohen, Sarah; Gauger, Eric; Fong, Allison; Gomez-Chiarri, Marta

2009-03-01

14

Quorum sensing regulates type III secretion in Vibrio harveyi and Vibrio parahaemolyticus.  

PubMed

In a process known as quorum sensing, bacteria communicate with one another by producing, releasing, detecting, and responding to signal molecules called autoinducers. Vibrio harveyi, a marine pathogen, uses two parallel quorum-sensing circuits, each consisting of an autoinducer-sensor pair, to control the expression of genes required for bioluminescence and a number of other target genes. Genetic screens designed to discover autoinducer-regulated targets in V. harveyi have revealed genes encoding components of a putative type III secretion (TTS) system. Using transcriptional reporter fusions and TTS protein localization studies, we show that the TTS system is indeed functional in V. harveyi and that expression of the genes encoding the secretion machinery requires an intact quorum-sensing signal transduction cascade. The newly completed genome of the closely related marine bacterium Vibrio parahaemolyticus, which is a human pathogen, shows that it possesses the genes encoding both of the V. harveyi-like quorum-sensing signaling circuits and that it also has a TTS system similar to that of V. harveyi. We show that quorum sensing regulates TTS in V. parahaemolyticus. Previous reports connecting quorum sensing to TTS in enterohemorrhagic and enteropathogenic Escherichia coli show that quorum sensing activates TTS at high cell density. Surprisingly, we find that at high cell density (in the presence of autoinducers), quorum sensing represses TTS in V. harveyi and V. parahaemolyticus. PMID:15175293

Henke, Jennifer M; Bassler, Bonnie L

2004-06-01

15

Molecular identification of Vibrio harveyi-related isolates associated with diseased aquatic organisms  

Microsoft Academic Search

Fifty strains belonging to Vibrio harveyi, Vibrio campbellii, and the recently described Vibrio rotiferianus, were analysed using phenotypic and genomic techniques with the aim of analysing the usefulness of the different techniques for the identification of V. harveyi-related species. The species V. harveyi and V. campbellii were phenotypically indistinguishable by more than 100 phenotypic features. Thirty-nine experimental strains were phenotypically

Bruno Gomez-Gil; Sonia Soto-Rodriguez; Alejandra Garcia-Gasca; Ana Roque; Ricardo Vazquez-Juarez; Fabiano L. Thompson; Jean Swings

2004-01-01

16

Identification of Vibrio harveyi isolated from diseased cultured wedge sole Dicologoglossa cuneata.  

PubMed

We report the first isolation of Vibrio harveyi from wedge sole Dicologoglossa cuneata. The pathogen was recovered from ulcers and internal organs of ailing cultured fish, from 7 different outbreaks between 2004 and 2006. The 15 isolates found were phenotypically characterized using biochemical tests and BIOLOG GN plates, which revealed high phenotypic diversity. Diagnosis was confirmed with PCR using V harveyi specific primers and partial 16S and 23S rRNA gene sequencing. A virulence evaluation of the isolates was also performed using fry and juvenile wedge sole. Significant mortalities were recorded by intraperitoneal injection; however, no mortalities were recorded by bath immersion. PMID:19565698

López, Jose R; de la Roca, Elena; Núñez, Soledad; de la Herran, Roberto; Navas, José I; Manchado, Manuel; Herrera, Marcelino; Toranzo, Alicia E

2009-04-27

17

Vibrio harveyi Associated with Aglaophenia octodonta (Hydrozoa, Cnidaria)  

Microsoft Academic Search

A previously unknown association between a luminous bacterium, Vibrio harveyi, and a benthic hydrozoan, Aglaophenia octodonta, is described. Aglaophenia hydrocladia showed a clear fluorescence in the folds along the hydrocaulus and at the base of the hydrotheca, suggesting the presence of luminous bacteria. This hypothesis was confirmed by isolation of luminous bacteria from Aglaophenia homogenates. Phenotypic characterization of bacterial isolates

L. Stabili; C. Gravili; S. Piraino; F. Boero; P. Alifano

2006-01-01

18

Interference of Cranberry Constituents in Cell–Cell Signaling System of Vibrio harveyi  

Microsoft Academic Search

Cranberry juice has long been recognized in folk medicine as a therapeutic agent, mainly in urinary track infections. It acts\\u000a as an antibiofilm agent against various pathogens. Quorum sensing is process where bacteria communicate with each other via\\u000a signal molecules known as autoinducers. This process is strongly involved in various bacterial pathological and physiological\\u000a pathways. Various strains of Vibrio harveyi

Mark Feldman; Ervin I. Weiss; Itzhak Ofek; Doron Steinberg

2009-01-01

19

Transcriptional regulation of lux genes transferred into Vibrio harveyi.  

PubMed Central

Past work has shown that transformed Escherichia coli is not a suitable vehicle for studying the expression and regulation of the cloned luminescence (lux) genes of Vibrio harveyi. Therefore, we have used a conjugative system to transfer lux genes cloned into E. coli back into V. harveyi, where they can be studied in the parental organism. To do this, lux DNA was inserted into a broad-spectrum vector, pKT230, cloned in E. coli, and then mobilized into V. harveyi by mating aided by the conjugative plasmid pRK2013, also contained in E. coli. Transfer of the wild-type luxD gene into the V. harveyi M17 mutant by this means resulted in complementation of the luxD mutation and full restoration of luminescence in the mutant; expression of transferase activity was induced if DNA upstream of luxC preceded the luxD gene on the plasmid, indicating the presence of a strong inducible promoter. To extend the usefulness of the transfer system, the gene for chloramphenicol acetyltransferase was inserted into the pKT230 vector as a reporter. The promoter upstream of luxC was verified to be cell density regulated and, in addition, glucose repressible. It is suggested that this promoter may be the primary autoregulated promoter of the V. harveyi luminescence system. Strong termination signals on both DNA strands were recognized and are located downstream from luxE at a point complementary to the longest mRNA from the lux operon. Structural lux genes transferred back into V. harveyi under control of the luxC promoter are expressed at very high levels in V. harveyi as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis: the gene transfer system is thus useful for expression of proteins as well as for studying the regulation of lux genes in their native environment. Images FIG. 4 FIG. 6 FIG. 8

Miyamoto, C M; Meighen, E A; Graham, A F

1990-01-01

20

Isolation of Vibrio harveyi bacteriophage with a potential for biocontrol of luminous vibriosis in hatchery environments  

Microsoft Academic Search

A double stranded DNA bacteriophage of Vibrio harveyi was isolated from shrimp farm water from the west coast of India. The isolated phage belonged to the family Siphoviridae. Its broad lytic activity against V. harveyi isolates both from the west and east coast of India suggested that it had a potential for biocontrol of luminous vibriosis due to V. harveyi

M. G. Vinod; M. M. Shivu; K. R. Umesha; B. C. Rajeeva; G. Krohne; Indrani Karunasagar; Iddya Karunasagar

2006-01-01

21

Expression of luciferases from Vibrio harveyi and Vibrio fischeri in filamentous cyanobacteria.  

PubMed Central

Shuttle vectors that had previously been shown to replicate both in Escherichia coli and in strains of Anabaena spp. were used to transfer the lux genes from Vibrio harveyi and Vibrio fischeri into Anabaena spp. The level of expression of luciferase in the cyanobacteria (up to 7,000 quanta cell-1 s-1) makes these genes good candidates for use as promoter probes during the differentiation of certain cells in a filament into heterocysts. Images

Schmetterer, G; Wolk, C P; Elhai, J

1986-01-01

22

Cloning and functional studies of a luxO regulator LuxT from Vibrio harveyi.  

PubMed

LuxO is the central regulator integrating the quorum sensing signals controlling autoinduction of luminescence in Vibrio harveyi. We have previously purified to homogeneity a new lux regulator, LuxT, that binds to the luxO promoter. Based on the sequence of the tryptic peptides of LuxT, degenerate oligonucleotides were designed for PCR of the genomic DNA. A 273 bp PCR DNA fragment containing sequences encoding the tryptic peptides was extended by inverse PCR to obtain the complete gene (luxT) encoding a protein of 153 amino acids which shares homology with the AcrR/TetR family of transcriptional regulators. The recombinant and native LuxT gave the same footprint binding between 117 and 149 bp upstream from the luxO initiation codon. Gene disruption of luxT in V. harveyi increased luxO expression and affected the cell density dependent induction of luminescence showing that LuxT was a repressor of luxO. As LuxT also affected the survival of the V. harveyi cells at high salt concentration and homologous proteins are present in other bacterial species, including the pathogen, Vibrio cholerae, the LuxT regulatory protein appears to be a general rather than a lux-specific regulator. PMID:11121579

Lin, Y H; Miyamoto, C; Meighen, E A

2000-12-01

23

Biosynthesis of myristic acid in luminescent bacteria. [Vibrio harveyi  

SciTech Connect

In vivo pulse-label studies have demonstrated that luminescent bacteria can provide myritic acid (14:0) required for the synthesis of the luciferase substrate myristyl aldehyde. Luminescent wild type Vibrio harveyi incubated with (/sup 14/C) acetate in a nutrient-depleted medium accumulated substantial tree (/sup 14/C)fatty acid (up to 20% of the total lipid label). Radio-gas chromatography revealed that > 75% of the labeled fatty acid is 14:0. No free fatty acid was detected in wild type cells labeled prior to the development of bioluminescence in the exponential growth phase, or in a dark mutant of V. harveyi (mutant M17) that requires exogenous 14:0 for light emission. The preferential accumulation of 14:0 was not observed when wild type cells were labeled with (/sup 14/C)acetate in regular growth medium. Moreover, all V. harveyi strains exhibited similar fatty acid mass compositions regardless of the state of bioluminescence. Since earlier work has shown that a luminescence-related acyltransferase (defective in the M17 mutant) can catalyze the deacylation of fatty acyl-acyl carrier protein in vitro, the present results are consistent with a model in which this enzyme diverts 14:0 to the luminescence system during fatty acid biosynthesis. Under normal conditions, the supply of 14:0 by this pathway is tightly regulated such that bioluminescence development does not significantly alter the total fatty acid composition.

Byers, D.M.

1987-05-01

24

Autoinducers Act as Biological Timers in Vibrio harveyi  

PubMed Central

Quorum sensing regulates cell density-dependent phenotypes and involves the synthesis, excretion and detection of so-called autoinducers. Vibrio harveyi strain ATCC BAA-1116 (recently reclassified as Vibrio campbellii), one of the best-characterized model organisms for the study of quorum sensing, produces and responds to three autoinducers. HAI-1, AI-2 and CAI-1 are recognized by different receptors, but all information is channeled into the same signaling cascade, which controls a specific set of genes. Here we examine temporal variations of availability and concentration of the three autoinducers in V. harveyi, and monitor the phenotypes they regulate, from the early exponential to the stationary growth phase in liquid culture. Specifically, the exponential growth phase is characterized by an increase in AI-2 and the induction of bioluminescence, while HAI-1 and CAI-1 are undetectable prior to the late exponential growth phase. CAI-1 activity reaches its maximum upon entry into stationary phase, while molar concentrations of AI-2 and HAI-1 become approximately equal. Similarly, autoinducer-dependent exoproteolytic activity increases at the transition into stationary phase. These findings are reflected in temporal alterations in expression of the luxR gene that encodes the master regulator LuxR, and of four autoinducer-regulated genes during growth. Moreover, in vitro phosphorylation assays reveal a tight correlation between the HAI-1/AI-2 ratio as input and levels of receptor-mediated phosphorylation of LuxU as output. Our study supports a model in which the combinations of autoinducers available, rather than cell density per se, determine the timing of various processes in V. harveyi populations.

Anetzberger, Claudia; Reiger, Matthias; Fekete, Agnes; Schell, Ursula; Stambrau, Nina; Plener, Laure; Kopka, Joachim; Schmitt-Kopplin, Phillippe; Hilbi, Hubert; Jung, Kirsten

2012-01-01

25

Immunoprotective analysis of VhhP2, a Vibrio harveyi vaccine candidate.  

PubMed

VhhP2 is an outer membrane protein identified in a pathogenic Vibrio harveyi strain, T4, isolated from diseased fish. When used as a subunit vaccine, purified recombinant VhhP2 affords high level of protection upon Japanese flounder against V. harveyi challenge. Vaccination with VhhP2 induced the expression of a number of immune-related genes, especially those encoding immunoglobulin M (IgM) and major histocompatibility complex (MHC) IIalpha. A VhhP2 surface display system, in the form of the fish commensal strain PF3 harboring the vhhP2-expressing plasmid pJVP, was constructed. PF3/pJVP is able to produce and present recombinant VhhP2 on cell surface. Vaccination of fish with live PF3/pJVP via intraperitoneal injection elicited strong immunoprotection. Vaccination of fish orally with live PF3/pJVP embedded in alginate microspheres also induced effective immunoprotection. In addition, a VhhP2-based surface display system was created, in which VhhP2 serves as a carrier for the surface delivery of a heterologous Edwardsiella tarda immunogen, Et18, that is fused in-frame to VhhP2. DH5alpha/pJVP18, which expresses and surface-displays the VhhP2-Et18 chimera, proved to be an effective vaccine that can protect fish against infections by V. harveyi and E. tarda to the extents comparable to those produced by vaccination with purified recombinant VhhP2 and Et18, respectively. These data suggest that VhhP2 may be applied as a vaccine and a vaccine carrier against infections by V. harveyi and other pathogens such as E. tarda. PMID:19428886

Sun, Kun; Zhang, Wei-Wei; Hou, Jin-Hui; Sun, Li

2009-05-11

26

Single cell analysis of Vibrio harveyi uncovers functional heterogeneity in response to quorum sensing signals  

PubMed Central

Background Vibrio harveyi and closely related species are important pathogens in aquaculture. A complex quorum sensing cascade involving three autoinducers controls bioluminescence and several genes encoding virulence factors. Single cell analysis of a V. harveyi population has already indicated intercellular heterogeneity in the production of bioluminescence. This study was undertaken to analyze the expression of various autoinducer-dependent genes in individual cells. Results Here we used reporter strains bearing promoter::gfp fusions to monitor the induction/repression of three autoinducer-regulated genes in wild type conjugates at the single cell level. Two genes involved in pathogenesis - vhp and vscP, which code for an exoprotease and a component of the type III secretion system, respectively, and luxC (the first gene in the lux operon) were chosen for analysis. The lux operon and the exoprotease gene are induced, while vscP is repressed at high cell density. As controls luxS and recA, whose expression is not dependent on autoinducers, were examined. The responses of the promoter::gfp fusions in individual cells from the same culture ranged from no to high induction. Importantly, simultaneous analysis of two autoinducer induced phenotypes, bioluminescence (light detection) and exoproteolytic activity (fluorescence of a promoter::gfp fusion), in single cells provided evidence for functional heterogeneity within a V. harveyi population. Conclusions Autoinducers are not only an indicator for cell density, but play a pivotal role in the coordination of physiological activities within the population.

2012-01-01

27

A bivalent Vibrio harveyi DNA vaccine induces strong protection in Japanese flounder (Paralichthys olivaceus).  

PubMed

Vibrio harveyi is a severe aquaculture pathogen that can infect a large number of fish species. In previous studies, we have identified two protective immunogens, DegQ and Vhp1, from pathogenic V. harveyi strains. In this study, we examined the potentials of DegQ and Vhp1 as monovalent and bivalent DNA vaccines in a Japanese flounder (Paralichthys olivaceus) model. For this purpose, the DNA vaccine plasmids pDegQ, pVhp1, and pDV were constructed. pDegQ and pVhp1 express, respectively, degQ and vhp1, while pDV expresses degQ and vhp1 as a bicistronic mRNA transcript (degQ-vhp1). Following vaccination, all vaccine plasmids were detected in muscle, spleen, and kidney at 7 and 14 days post-vaccination. At the same time points and in the same tissues, degQ, vhp1, and degQ-vhp1 mRNA transcripts were detected in, respectively, pDegQ-, pVhp1-, and pDV-vaccinated fish. Production of DegQ and Vhp1 proteins was found in the muscle tissues of pDegQ- and pVhp1-vaccinated fish respectively, while both DegQ and Vhp1 proteins were detected in pDV-vaccinated fish. Compared to control fish, fish vaccinated with pDegQ and pVhp1 exhibited, respectively, 64.1% and 56.5% relative percent survival rates following lethal V. harveyi challenge, while fish vaccinated with pDV exhibited a survival rate that is more than 20% higher than those induced by pDegQ and pVhp1. Furthermore, in addition to V. harveyi, pDV also elicited dramatic cross protection against Vibrio parahaemolyticus. Immunological analysis showed that all vaccine plasmids induced the production of specific serum antibodies and the expression of a wide range of immune genes; however, the induction folds effected by pDV were in general higher than those effected by pDegQ and pVhp1. These results indicate that DNA vaccines, such as that represented by pDV, is a good vaccination option to combat V. harveyi-related vibriosis. PMID:21513763

Hu, Yong-hua; Sun, Li

2011-06-10

28

Tryptophan biosynthesis in the marine luminous bacterium Vibrio harveyi.  

PubMed Central

Tryptophan biosynthetic enzyme levels in wild-type Vibrio harveyi and a number of tryptophan auxotrophs of this species were coordinately regulated over a 100-fold range of specific activities. The tryptophan analog indoleacrylic acid evoked substantial derepression of the enzymes in wild-type cells. Even higher enzyme levels were attained in auxotrophs starved for tryptophan, regardless of the location of the block in the pathway. A derepressed mutant selected by resistance to 5-fluorotryptophan was found to have elevated basal levels of trp gene expression; these basal levels were increased only two- to threefold by tryptophan limitation. The taxonomic implications of these and other biochemical results support previous suggestions that the marine luminous bacteria are more closely related to enteric bacteria than to other gram-negative taxa.

Bieger, C D; Crawford, I P

1983-01-01

29

Sequencing and preliminary characterization of the Na+-translocating NADH:ubiquinone oxidoreductase from Vibrio harveyi.  

PubMed

The Na(+)-translocating NADH: ubiquinone oxidoreductase (Na(+)-NQR) generates an electrochemical Na(+) potential driven by aerobic respiration. Previous studies on the enzyme from Vibrio alginolyticus have shown that the Na(+)-NQR has six subunits, and it is known to contain FAD and an FeS center as redox cofactors. In the current work, the enzyme from the marine bacterium Vibrio harveyi has been purified and characterized. In addition to FAD, a second flavin, tentatively identified as FMN, was discovered to be covalently attached to the NqrC subunit. The purified V. harveyi Na(+)-NQR was reconstituted into proteoliposomes. The generation of a transmembrane electric potential by the enzyme upon NADH:Q(1) oxidoreduction was strictly dependent on Na(+), resistant to the protonophore CCCP, and sensitive to the sodium ionophore ETH-157, showing that the enzyme operates as a primary electrogenic sodium pump. Interior alkalinization of the inside-out proteoliposomes due to the operation of the Na(+)-NQR was accelerated by CCCP, inhibited by valinomycin, and completely arrested by ETH-157. Hence, the protons required for ubiquinol formation must be taken up from the outside of the liposomes, which corresponds to the bacterial cytoplasm. The Na(+)-NQR operon from this bacterium was sequenced, and the sequence shows strong homology to the previously reported Na(+)-NQR operons from V. alginolyticus and Haemophilus influenzae. Homology studies show that a number of other bacteria, including a number of pathogenic species, also have an Na(+)-NQR operon. PMID:10587447

Zhou, W; Bertsova, Y V; Feng, B; Tsatsos, P; Verkhovskaya, M L; Gennis, R B; Bogachev, A V; Barquera, B

1999-12-01

30

Mechanisms underlying the additive and redundant Qrr phenotypes in Vibrio harveyi and Vibrio cholerae.  

PubMed

Vibrio harveyi and Vibrio cholerae regulate their virulence factors according to the local cell-population density in a regulatory system called quorum sensing. Their quorum sensing systems contain a small RNA (sRNA) circuit to regulate expression of a master transcriptional regulator via multiple quorum regulated RNA (Qrr) and a protein chaperon Hfq. Experiments and genetic analysis show that their respective quorum sensing networks are topologically equivalent and have homologous components, yet they respond differently to the same experimental conditions. In particular, V. harveyi Qrr are additive because all of its Qrr are required to maintain wild-type-like repression of its master transcriptional regulator. Conversely, V. cholerae Qrr are redundant because any of its Qrr is sufficient to repress its master transcriptional regulator. Given the striking similarities between their quorum sensing systems, experimentalists have been unable to identify conclusively the mechanisms behind these phenotypic differences. Nevertheless, the current hypothesis in the literature is that dosage compensation is the mechanism underlying redundancy. In this work, we identify the mechanisms underlying Qrr redundancy using a detailed mathematical model of the V. harveyi and V. cholerae sRNA circuits. We show that there are exactly two different cases underlying Qrr redundancy and that dosage compensation is unnecessary and insufficient to explain Qrr redundancy. Although V. harveyi Qrr are additive when the perturbations in Qrr are large, we predict that V. harveyi and V. cholerae Qrr are redundant when the perturbations in Qrr are small. We argue that the additive and redundant Qrr phenotypes can emerge from parametric differences in the sRNA circuit. In particular, we find that the affinity of Qrr and its expression relative to the master transcriptional regulator determine the level of redundancy in V. harveyi and V. cholerae. Furthermore, the additive and redundant Qrr phenotypes reflect differences in the concentration of Hfq-Qrr in V. harveyi and V. cholerae. We use our model to test the dosage compensation hypothesis and show that decreasing the expression of qrr, rather than removing dosage compensation, abolishes Qrr redundancy in V. cholerae. Further experimentation is needed to test our results and both Qrr redundancy hypotheses. PMID:24018202

Hunter, Geoffrey A M; Keener, James P

2014-01-01

31

Association of Vibrio harveyi with mortalities in cultured marine fish in Kuwait  

Microsoft Academic Search

A Vibrio sp. frequently associated with mortalities in cultured silvery black porgy Acanthopagrus cuvieri and brown-spotted grouper Epinephelus tauvina was studied. Silvery black porgy was susceptible to laboratory infection with this isolate only when administered intramuscularly (IM), but the isolate could infect brown-spotted grouper by both IM and intraperitoneal (IP) routes. The isolate was identified as Vibrio harveyi. The 5

M. O. Saeed

1995-01-01

32

Vibrio harveyi quorum sensing: a coincidence detector for two autoinducers controls gene expression  

PubMed Central

In a process called quorum sensing, bacteria communicate with one another by exchanging chemical signals called autoinducers. In the bioluminescent marine bacterium Vibrio harveyi, two different auto inducers (AI-1 and AI-2) regulate light emission. Detection of and response to the V.harveyi autoinducers are accomplished through two two-component sensory relay systems: AI-1 is detected by the sensor LuxN and AI-2 by LuxPQ. Here we further define the V.harveyi quorum-sensing regulon by identifying 10 new quorum-sensing-controlled target genes. Our examination of signal processing and integration in the V.harveyi quorum-sensing circuit suggests that AI-1 and AI-2 act synergistically, and that the V.harveyi quorum-sensing circuit may function exclusively as a ‘coincidence detector’ that discriminates between conditions in which both autoinducers are present and all other conditions.

Mok, Kenny C.; Wingreen, Ned S.; Bassler, Bonnie L.

2003-01-01

33

Chitoporin from Vibrio harveyi, a Channel with Exceptional Sugar Specificity  

PubMed Central

Chitoporin (VhChiP) is a sugar-specific channel responsible for the transport of chitooligosaccharides through the outer membrane of the marine bacterium Vibrio harveyi. Single channel reconstitution into black lipid membrane allowed single chitosugar binding events in the channel to be resolved. VhChiP has an exceptionally high substrate affinity, with a binding constant of K = 5.0 × 106 m?1 for its best substrate (chitohexaose). The on-rates of chitosugars depend on applied voltages, as well as the side of the sugar addition, clearly indicating the inherent asymmetry of the VhChiP lumen. The binding affinity of VhChiP for chitohexaose is 1–5 orders of magnitude larger than that of other known sugar-specific porins for their preferred substrates. Thus, VhChiP is the most potent sugar-specific channel reported to date, with its high efficiency presumably reflecting the need for the bacterium to take up chitin-containing nutrients promptly under turbulent aquatic conditions to exploit them efficiently as its sole source of energy.

Suginta, Wipa; Chumjan, Watcharin; Mahendran, Kozhinjampara R.; Schulte, Albert; Winterhalter, Mathias

2013-01-01

34

Proteomic analysis of differentially expressed proteins in Penaeus monodon hemocytes after Vibrio harveyi infection  

PubMed Central

Background Viral and bacterial diseases can cause mass mortalities in commercial shrimp aquaculture. In contrast to studies on the antiviral response, the responses of shrimps to bacterial infections by high throughput techniques have been reported only at the transcriptional level and not at the translational level. In this study, a proteomic analysis of shrimp hemocytes to identify differentially expressed proteins in response to a luminous bacterium Vibrio harveyi was evaluated for its feasibility and is reported for the first time. Results The two-dimensional gel electrophoresis (2-DE) patterns of the hemocyte proteins from the unchallenged and V. harveyi challenged shrimp, Penaeus monodon, at 24 and 48 h post infection were compared. From this, 27 differentially expressed protein spots, and a further 12 weakly to non-differentially regulated control spots, were selected for further analyses by the LC-ESI-MS/MS. The 21 differentially expressed proteins that could be identified by homologous annotation were comprised of proteins that are directly involved in the host defense responses, such as hemocyanin, prophenoloxidase, serine proteinase-like protein, heat shock protein 90 and alpha-2-macroglobulin, and those involved in signal transduction, such as the14-3-3 protein epsilon and calmodulin. Western blot analysis confirmed the up-regulation of hemocyanin expression upon bacterial infection. The expression of the selected proteins which were the representatives of the down-regulated proteins (the 14-3-3 protein epsilon and alpha-2-macroglobulin) and of the up-regulated proteins (hemocyanin) was further assessed at the transcription level using real-time RT-PCR. Conclusions This work suggests the usefulness of a proteomic approach to the study of shrimp immunity and revealed hemocyte proteins whose expression were up regulated upon V. harveyi infection such as hemocyanin, arginine kinase and down regulated such as alpha-2-macroglobulin, calmodulin and 14-3-3 protein epsilon. The information is useful for understanding the immune system of shrimp against pathogenic bacteria.

2010-01-01

35

Vibrio harveyi Nitroreductase Is Also a Chromate Reductase  

PubMed Central

The chromate reductase purified from Pseudomonas ambigua was found to be homologous with several nitroreductases. Escherichia coli DH5? and Vibrio harveyi KCTC 2720 nitroreductases were chosen for the present study, and their chromate-reducing activities were determined. A fusion between glutathione S-transferase (GST) and E. coli DH5? NfsA (GST-EcNfsA), a fusion between GST and E. coli DH5? NfsB (GST-EcNfsB), and a fusion between GST and V. harveyi KCTC 2720 NfsA (GST-VhNfsA) were prepared for their overproduction and easy purification. GST-EcNfsA, GST-EcNFsB, and GST-VhNFsA efficiently reduced nitrofurazone and 2,4,6-trinitrotoluene (TNT) as their nitro substrates. The Km values for GST-EcNfsA, GST-EcNfsB, and GST-VhNfsA for chromate reduction were 11.8, 23.5, and 5.4 ?M, respectively. The Vmax values for GST-EcNfsA, GST-EcNfsB, and GST-VhNfsA were 3.8, 3.9, and 10.7 nmol/min/mg of protein, respectively. GST-VhNfsA was the most effective of the three chromate reductases, as determined by each Vmax/Km value. The optimal temperatures of GST-EcNfsA, GST-EcNfsB, and GST-VhNfsA for chromate reduction were 55, 30, and 30°C, respectively. Thus, it is confirmed that nitroreductase can also act as a chromate reductase. Nitroreductases may be used in chromate remediation. GST-EcNfsA, GST-EcNfsB, and GST-VhNfsA have a molecular mass of 50 kDa and exist as a monomer in solution. Thin-layer chromatography showed that GST-EcNfsA, GST-EcNfsB, and GST-VhNfsA contain FMN as a cofactor. GST-VhNfsA reduced Cr(VI) to Cr(III). Cr(III) was much less toxic to E. coli than Cr(VI).

Kwak, Young Hak; Lee, Dong Seok; Kim, Han Bok

2003-01-01

36

Virulence Changes to Harveyi Clade Bacteria Infected with Bacteriophage from Vibrio owensii.  

PubMed

Vibrio owensii is one of the most virulent vibrios known being able to kill crustacean larvae at 10(2) CFU ml(-1). This study describes virulence changes to naïve strains of Vibrio harveyi and Vibrio campbellii when infected with the bacteriophage VOB from a closely related species V. owensii 47666-1. The bacteriophage from V. owensii was induced into lytic phase by using mitomycin C at 100 ng ml(-1). One strain of V. harveyi and two strains of V. campbellii from 29 tested containing no prophage were susceptible to lysogenic conversion with VOB. Virulence changes induced in Harveyi clade bacteria included the up-regulation of protein secretion, statistically significant increased haemolysin and chitinase production and increased mortality to nauplii of Penaeus monodon. No change in siderophore production was observed. Bacteriophage VOB is likely to be responsible for some of the virulence factors expressed by V. owensii. As this bacteriophage is able to infect strains of V. harveyi and V. campbellii this phage may contribute to increased virulence of other vibrios in aquaculture and in the natural environment. PMID:24426274

Busico-Salcedo, Nancy; Owens, Leigh

2013-09-01

37

Unexpected photoreactivation of Vibrio harveyi bacteria living in ionization environment  

NASA Astrophysics Data System (ADS)

Bacteria undergoing environmental effects is extremely interesting for structural, mechanistic, and evolutionary implications. Luminescent bacteria that have evolved in a specific ambient have developed particular responses and their behavior can give us new suggestions on the task and production of luciferina proteins. To analyze the UV interaction under controlled laboratory conditions, we used photoluminescent bacterial strains belonging to a new species evolutionarily close to Vibrio harveyi sampled from a coastal cave with a high radon content that generates ionizing radiation. The survival of the bacterial strains was analyzed, in the light and in the dark, following a variety of genotoxic treatments including UV radiation exposure. The strains were irradiated by a germicide lamp. The results demonstrated that most of the strains exhibited a low rate of survival after the UV exposure. After irradiation by visible light following the UV exposure, all strains showed a high capability of photoreactivation when grown. This capability was quite unexpected because these bacteria were sampled from a dark ambient without UV radiation. This leads us to hypothesize that the photoreactivation in these bacteria might have been evolved to repair DNA lesions also induced by different radiation sources other than UV (e.g., x-ray) and that the luminescent bacteria might use their own light emission to carry out the photoreactivation. The high capability of photoreactivation of these bacteria was also justified by the results of deconvolution. The deconvolution was applied to the emission spectra and it was able to show evidence of different light peaks. The presence of the visible peak could control the photolysis enzyme.

Alifano, P.; Nassisi, V.; Siciliano, M. V.; Talà, A.; Tredici, S. M.

2011-05-01

38

The Small RNA Chaperone Hfq and Multiple Small RNAs Control Quorum Sensing in Vibrio harveyi and Vibrio cholerae  

Microsoft Academic Search

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

Derrick H. Lenz; Kenny C. Mok; Brendan N. Lilley; Rahul V. Kulkarni; Ned S. Wingreen; Bonnie L. Bassler

2004-01-01

39

Elongation of exogenous fatty acids by the bioluminescent bacterium Vibrio harveyi  

Microsoft Academic Search

Bioluminescent bacteria require myristic acid (C14:0) to produce the myristaldehyde substrate of the light-emitting luciferase reaction. Since both endogenous and exogenous C14:0 can be used for this purpose, the metabolism of exogenous fatty acids by luminescent bacteria has been investigated. Both Vibrio harveyi and Vibrio fischeri incorporated label from (1-14C)myristic acid (C14:0) into phospholipid acyl chains as well as into

1989-01-01

40

Bacterial Bioluminescence: Isolation and Expression of the Luciferase Genes from Vibrio harveyi  

Microsoft Academic Search

Genes for the luciferase enzyme of Vibrio harveyi were isolated in Escherichia coli by a general method in which nonluminous, transposon insertion mutants were used. Conditions necessary for light production in E. coli were examined. Stimulation of transcription of the genes for luciferase (lux A and lux B) was required for efficient synthesis of luciferase. To enhance transcription bacteriophage promoter

Robert Belas; Alan Mileham; Daniel Cohn; Marcia Hilmen; Melvin Simon; Michael Silverman

1982-01-01

41

Identification of upregulated immune-related genes in Vibrio harveyi challenged Penaeus monodon postlarvae  

Microsoft Academic Search

A subtracted cDNA library was constructed and analyzed to elucidate the response of Penaeus monodon postlarvae challenged with Vibrio harveyi. As many as 960 randomly selected cDNA fragments generated through suppression subtractive hybridization were single pass sequenced. Forty five genes and 20 hypothetical proteins were identified, a few being first reports from shrimps. The most abundant immune relevant genes were

S. Nayak; S. K. Singh; N. Ramaiah; R. A. Sreepada

2010-01-01

42

Detection of biohazardous materials in water by measuring bioluminescence reduction with the marine organism Vibrio harveyi  

Microsoft Academic Search

This study evaluated two bioassay methods, direct and growth, using Vibrio harveyi, a bioluminescent bacterium, to detect biohazardous materials in water. The end point measured for the evaluation of the toxicity of the various substances tested was the median effective concentration for bioluminescence reduction. Thirty?four compounds were tested, including representatives from the following chemical categories: azide, alcohols, antibiotics, antioxidants, detergents,

Kenneth W. Thomulka; David J. McGee; John H. Lange

1993-01-01

43

Genetic Organization of the Vibrio harveyi dnaA Gene Region and Analysis of the Function of the V. harveyi DnaA Protein in Escherichia coli  

Microsoft Academic Search

The Vibrionaceae family is distantly related to Enterobacteriaceae within the group of bacteria possessing the Dam methylase system. We have cloned, sequenced, and analyzed the dnaA gene region of Vibrio harveyi and found that although the organization of the V. harveyi dnaA region differs from that of Escherichia coli, the expression of both genes is autoregulated and ATP-DnaA binds cooperatively

Dvora Berenstein; Kirsten Olesen; Christian Speck; Ole Skovgaard

2002-01-01

44

Metabolic profiling of the tissue-specific responses in mussel Mytilus galloprovincialis towards Vibrio harveyi challenge.  

PubMed

Mussel Mytilus galloprovincialis is a marine aquaculture shellfish distributing widely along the coast in north China. In this work, we studied the differential metabolic responses induced by Vibrio harveyi in digestive gland and gill tissues from M. galloprovincialis using NMR-based metabolomics. The differential metabolic responses in the two tissue types were detected, except the similarly altered taurine and betaine. These metabolic responses suggested that V. harveyi mainly induced osmotic disruption and reduced energy demand via the metabolic pathways of glucose synthesis and ATP/AMP conversion in mussel digestive gland. In mussel gill tissues, V. harveyi basically caused osmotic stress and possible reduced energy demand as shown by the elevated phosphocholine that is involved in one of the metabolic pathways of ATP synthesis from ADP and phosphocholine. The altered mRNA expression levels of related genes (superoxide dismutase with copper and zinc, heat shock protein 90, defensin and lysozyme) suggested that V. harveyi induced clear oxidative and immune stresses in both digestive gland and gill tissues. However, the mRNA expression levels of both lysozyme and defensin in digestive gland were more significantly up-regulated than those in gill from V. harveyi-challenged mussel M. galloprovincialis, meaning that the immune organ, digestive gland, was more sensitive than gill. Overall, our results indicated that V. harveyi could induce tissue-specific metabolic responses in mussel M. galloprovincialis. PMID:24911264

Liu, Xiaoli; Ji, Chenglong; Zhao, Jianmin; Wang, Qing; Li, Fei; Wu, Huifeng

2014-08-01

45

VanT, a Homologue of Vibrio harveyi LuxR, Regulates Serine, Metalloprotease, Pigment, and Biofilm Production in Vibrio anguillarum  

Microsoft Academic Search

Vibrio anguillarum possesses at least two N-acylhomoserine lactone (AHL) quorum-sensing circuits, one of which is related to the luxMN system of Vibrio harveyi. In this study, we have cloned an additional gene of this circuit, vanT, encoding a V. harveyi LuxR-like transcriptional regulator. A V. anguillarum vanT null mutation resulted in a significant decrease in total protease activity due to

Antony Croxatto; Victoria J. Chalker; Johan Lauritz; Jana Jass; Andrea Hardman; Paul Williams; M. Camara; Debra L. Milton

2002-01-01

46

Novel ?-N-acetylglucosaminidases from Vibrio harveyi 650: Cloning, expression, enzymatic properties, and subsite identification  

Microsoft Academic Search

BACKGROUND: Since chitin is a highly abundant natural biopolymer, many attempts have been made to convert this insoluble polysaccharide into commercially valuable products using chitinases and ?-N-acetylglucosaminidases (GlcNAcases). We have previously reported the structure and function of chitinase A from Vibrio harveyi 650. This study t reports the identification of two GlcNAcases from the same organism and their detailed functional

Wipa Suginta; Duangkamon Chuenark; Mamiko Mizuhara; Tamo Fukamizo

2010-01-01

47

Elongation of exogenous fatty acids by the bioluminescent bacterium Vibrio harveyi  

SciTech Connect

Bioluminescent bacteria require myristic acid (C14:0) to produce the myristaldehyde substrate of the light-emitting luciferase reaction. Since both endogenous and exogenous C14:0 can be used for this purpose, the metabolism of exogenous fatty acids by luminescent bacteria has been investigated. Both Vibrio harveyi and Vibrio fischeri incorporated label from (1-14C)myristic acid (C14:0) into phospholipid acyl chains as well as into CO2. In contrast, Photobacterium phosphoreum did not exhibit phospholipid acylation or beta-oxidation using exogenous fatty acids. Unlike Escherichia coli, the two Vibrio species can directly elongate fatty acids such as octanoic (C8:0), lauric (C12:0), and myristic acid, as demonstrated by radio-gas liquid chromatography. The induction of bioluminescence in late exponential growth had little effect on the ability of V. harveyi to elongate fatty acids, but it did increase the amount of C14:0 relative to C16:0 labeled from (14C)C8:0. This was not observed in a dark mutant of V. harveyi that is incapable of supplying endogenous C14:0 for luminescence. Cerulenin preferentially decreased the labeling of C16:0 and of unsaturated fatty acids from all 14C-labeled fatty acid precursors as well as from (14C)acetate, suggesting that common mechanisms may be involved in elongation of fatty acids from endogenous and exogenous sources. Fatty acylation of the luminescence-related synthetase and reductase enzymes responsible for aldehyde synthesis exhibited a chain-length preference for C14:0, which also was indicated by reverse-phase thin-layer chromatography of the acyl groups attached to these enzymes. The ability of V. harveyi to activate and elongate exogenous fatty acids may be related to an adaptive requirement to metabolize intracellular C14:0 generated by the luciferase reaction during luminescence development.

Byers, D.M.

1989-01-01

48

Elongation of exogenous fatty acids by the bioluminescent bacterium Vibrio harveyi.  

PubMed Central

Bioluminescent bacteria require myristic acid (C14:0) to produce the myristaldehyde substrate of the light-emitting luciferase reaction. Since both endogenous and exogenous C14:0 can be used for this purpose, the metabolism of exogenous fatty acids by luminescent bacteria has been investigated. Both Vibrio harveyi and Vibrio fischeri incorporated label from [1-14C]myristic acid (C14:0) into phospholipid acyl chains as well as into CO2. In contrast, Photobacterium phosphoreum did not exhibit phospholipid acylation or beta-oxidation using exogenous fatty acids. Unlike Escherichia coli, the two Vibrio species can directly elongate fatty acids such as octanoic (C8:0), lauric (C12:0), and myristic acid, as demonstrated by radio-gas liquid chromatography. The induction of bioluminescence in late exponential growth had little effect on the ability of V. harveyi to elongate fatty acids, but it did increase the amount of C14:0 relative to C16:0 labeled from [14C]C8:0. This was not observed in a dark mutant of V. harveyi that is incapable of supplying endogenous C14:0 for luminescence. Cerulenin preferentially decreased the labeling of C16:0 and of unsaturated fatty acids from all 14C-labeled fatty acid precursors as well as from [14C]acetate, suggesting that common mechanisms may be involved in elongation of fatty acids from endogenous and exogenous sources. Fatty acylation of the luminescence-related synthetase and reductase enzymes responsible for aldehyde synthesis exhibited a chain-length preference for C14:0, which also was indicated by reverse-phase thin-layer chromatography of the acyl groups attached to these enzymes. The ability of V. harveyi to activate and elongate exogenous fatty acids may be related to an adaptive requirement to metabolize intracellular C14:0 generated by the luciferase reaction during luminescence development. Images

Byers, D M

1989-01-01

49

Genetic Organization of the Vibrio harveyi dnaA Gene Region and Analysis of the Function of the V. harveyi DnaA Protein in Escherichia coli  

PubMed Central

The Vibrionaceae family is distantly related to Enterobacteriaceae within the group of bacteria possessing the Dam methylase system. We have cloned, sequenced, and analyzed the dnaA gene region of Vibrio harveyi and found that although the organization of the V. harveyi dnaA region differs from that of Escherichia coli, the expression of both genes is autoregulated and ATP-DnaA binds cooperatively to ATP-DnaA boxes in the dnaA promoter region. The DnaA proteins of V. harveyi and E. coli are interchangeable and function nearly identically in controlling dnaA transcription and the initiation of chromosomal DNA replication despite the evolutionary distance between these bacteria.

Berenstein, Dvora; Olesen, Kirsten; Speck, Christian; Skovgaard, Ole

2002-01-01

50

Identification of upregulated immune-related genes in Vibrio harveyi challenged Penaeus monodon postlarvae.  

PubMed

A subtracted cDNA library was constructed and analyzed to elucidate the response of Penaeus monodon postlarvae challenged with Vibrio harveyi. As many as 960 randomly selected cDNA fragments generated through suppression subtractive hybridization were single pass sequenced. Forty five genes and 20 hypothetical proteins were identified, a few being first reports from shrimps. The most abundant immune relevant genes were ferritin, hemocyanin, and TCTP (translationally controlled tumor protein) indicating their upregulation as also confirmed through qPCR. Post-infection qPCR analyses confirmed 2.04, 2.09, 3.28, 5.49, 6.47, and 11.63 fold rise respectively in ferritin, penaeidin, MnSOD, lysozyme, TCTP, and hemocyanin genes. These genes may be involved in the regulation of the host defense against V. harveyi. PMID:20580834

Nayak, S; Singh, S K; Ramaiah, N; Sreepada, R A

2010-09-01

51

The Vibrio harveyi quorum-sensing system uses shared regulatory components to discriminate between multiple autoinducers.  

PubMed

The quorum-sensing bacterium Vibrio harveyi produces and responds to three autoinducers (AIs), and this sensory information converges to control the expression of bioluminescence, biofilm formation, type III secretion (TTS), and protease production. The AIs are detected by cognate sensor histidine kinases that all relay phosphate to the shared response regulator LuxO. LuxO indirectly represses the master regulator of quorum sensing, LuxR, through the activation of multiple genes encoding small regulatory RNAs (called qrr genes for Quorum Regulatory RNA). Here we use differential fluorescence induction to identify 50 quorum-sensing-controlled promoters. Some promoters only showed significant responses in the simultaneous presence of all three AIs, while others displayed substantial responses to the individual AIs. A differential response to each AI input state was also observed for qrr and luxR expression and LuxR protein production. Individual cell analyses revealed that, in each case, all the bacteria in the population respond in unison to the various AI inputs. We propose that the V. harveyi quorum-sensing transition is not switch-like but rather operates in a graded manner, and that this signaling arrangement, which uses shared regulatory proteins, nonetheless provides V. harveyi a mechanism to respond uniquely to different AI input states. PMID:17015436

Waters, Christopher M; Bassler, Bonnie L

2006-10-01

52

Translocation of Vibrio harveyi N,N'-diacetylchitobiase to the outer membrane of Escherichia coli.  

PubMed Central

The gene encoding N,N'-diacetylchitobiase (chitobiase) of the chitinolytic marine bacterium Vibrio harveyi has been isolated. While expression of the chitobiase gene (chb) was inducible by N,N'-diacetylchitobiose in V. harveyi, it was expressed constitutively when cloned in Escherichia coli, suggesting that controlling elements are not closely linked to chb. Chitobiase was found in the membrane fraction of E. coli cells containing plasmids with the cloned V. harveyi chb gene. When membranes of such cells were separated on Osborn gradients, chitobiase activity was found mainly in the outer membrane band. Translocation of the enzyme to the outer membrane was accompanied by cleavage of a signal peptide. A fusion protein, in which 22 amino acids from the amino terminus of prechitobiase were replaced with 21 amino acids from the pUC19 lacZ amino terminus, was not processed, and 99% of the activity was located in the cytoplasmic fraction. A homology to six amino acids surrounding the lipoprotein processing and modification site was found near the amino terminus of prechitobiase. Images

Jannatipour, M; Soto-Gil, R W; Childers, L C; Zyskind, J W

1987-01-01

53

Expression of bioluminescence by Escherichia coli containing recombinant Vibrio harveyi DNA.  

PubMed Central

When isogenic strains of Escherichia coli, RR1 (rec+) and HB101 (recA), were transformed with mapped recombinant plasmids known to contain Vibrio harveyi luciferase genes and large regions of DNA flanking on both sides, a small percentage (0.005%) of the colonies expressed high levels of luminescence (up to 10(12) quanta s-1 ml-1) in the absence of added aldehyde. The altered ability to express light was found to be due to a mutation in the host and not to an alteration in the recombinant DNA. When these bright colonies were cured of plasmid, they could be retransformed with cloned V. harveyi gene fragments in cis and in trans to yield luminescent colonies at 100% frequency. The maximum length of V. harveyi DNA required to produce light-emitting E. coli was shorter (6.3 kilobase pairs) than that required for expression of the V. fischeri system in E. coli. Cell extracts from bright clones contained wild-type levels of activity for the heteropolymeric (alpha beta) luciferase; fatty acid labeling revealed the presence of the three acylated polypeptides of the fatty acid reductase system which is involved in aldehyde biosynthesis for the luminescence reaction. The increased light emission in the mutant bacteria appeared to arise in part from production of higher levels of polycistronic mRNAs coding for luciferase. Images

Miyamoto, C; Byers, D; Graham, A F; Meighen, E A

1987-01-01

54

Differential regulation of enzyme activities involved in aldehyde metabolism in the luminescent bacterium Vibrio harveyi.  

PubMed Central

The effects of catabolite repression and nutrient abundance on the activities of Vibrio harveyi enzymes known to be related to aldehyde metabolism were investigated. The growth of cells in complex medium containing glucose, which decreases in vivo luminescence and luciferase synthesis, also resulted in decreases in the specific activities of V. harveyi aldehyde dehydrogenase and acyl carrier protein acyltransferase as well as in the degree of fatty acylation of three bioluminescence-specific polypeptides (32, 42, and 57 kilodaltons), as monitored by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. This repression was partially alleviated in glucose medium containing cyclic AMP. The acylation of the above-mentioned proteins, in addition to light emission and luciferase and acyltransferase activities, was also repressed when cells were grown in minimal medium, with partial recovery of these functions upon the addition of arginine. In contrast, aldehyde dehydrogenase activity was increased in minimal medium. These results suggest that the 42-, 57-, and 32-kilodalton proteins, which are responsible for the supply and reduction of fatty acids to form aldehydes for the luciferase reaction, are regulated in the same way as luciferase under the above-described conditions. However, aldehyde dehydrogenase, whose role in V. harveyi aldehyde metabolism is not yet known, is regulated in a different way with respect to nutrient composition. Images

Byers, D M; Bognar, A; Meighen, E A

1988-01-01

55

Polycistronic mRNAs code for polypeptides of the Vibrio harveyi luminescence system  

SciTech Connect

DNA coding for the ..cap alpha.. and ..beta.. subunits of Vibrio harveyi luciferase, the luxA and luxB genes, and the adjoining chromosomal regions on both sides of these genes (total of 18 kilobase pairs) was cloned into Escherichia coli. Using labeled DNA coding for the ..cap alpha.. subunit as a hybridization probe, the authors identified a set of polycistronic mRNAs (2.6, 4, 7, and 8 kilobases) by Northern blotting; the most prominent of these was the one 4 kilobases long. This set of mRNAs was induced during the development of bioluminescence in V. harveyi. Furthermore, the same set of mRNAs was synthesized in E. coli by a recombinant plasmid that contained a 12-kilobase pair length of V. harveyi DNA and expressed the genes for the luciferase subunits. A cloned DNA segment corresponding to the major 4-kilobase mRNA coded for the ..cap alpha.. and ..beta.. subunits of luciferase, as well as a 32,000-dalton protein upstream from these genes that could be specifically modified by acyl-coenzyme A and is a component of the bioluminescence system. V. harveyi mRNA that was hybridized to the released from cloned DNA encompassing the luxA and luxB genes was translated in vitro. Luciferase ..cap alpha.. and ..beta.. subunits and the 32,000-dalton polypeptide were detected among the products, along with 42,000- and 55,000-dalton polypeptides, which are encoded downstream from the lux genes and are thought to be involved in luminescence.

Miyamoto, C.M.; Graham, A.D.; Boylan, M.; Evans, J.F.; Hasel, K.W.; Meighen, E.A.; Graham, A.F.

1985-03-01

56

Polycistronic mRNAs code for polypeptides of the Vibrio harveyi luminescence system.  

PubMed Central

DNA coding for the alpha and beta subunits of Vibrio harveyi luciferase, the luxA and luxB genes, and the adjoining chromosomal regions on both sides of these genes (total of 18 kilobase pairs) was cloned into Escherichia coli. Using labeled DNA coding for the alpha subunit as a hybridization probe, we identified a set of polycistronic mRNAs (2.6, 4, 7, and 8 kilobases) by Northern blotting; the most prominent of these was the one 4 kilobases long. This set of mRNAs was induced during the development of bioluminescence in V. harveyi. Furthermore, the same set of mRNAs was synthesized in E. coli by a recombinant plasmid that contained a 12-kilobase pair length of V. harveyi DNA and expressed the genes for the luciferase subunits. A cloned DNA segment corresponding to the major 4-kilobase mRNA coded for the alpha and beta subunits of luciferase, as well as a 32,000-dalton protein upstream from these genes that could be specifically modified by acyl-coenzyme A and is a component of the bioluminescence system. V. harveyi mRNA that was hybridized to and released from cloned DNA encompassing the luxA and luxB genes was translated in vitro. Luciferase alpha and beta subunits and the 32,000-dalton polypeptide were detected among the products, along with 42,000- and 55,000-dalton polypeptides, which are encoded downstream from the lux genes and are thought to be involved in luminescence. Images

Miyamoto, C M; Graham, A D; Boylan, M; Evans, J F; Hasel, K W; Meighen, E A; Graham, A F

1985-01-01

57

Profiling of a few immune responsive genes expressed in postlarvae of Fenneropenaeus indicus challenged with Vibrio harveyi D3  

Microsoft Academic Search

We identified 38 genes and eight hypothetical proteins by sequencing of 1200 clones from a Vibrio harveyi challenged Fenneropenaeus indicus subtracted cDNA library. Based on physiological roles and functions these genes were categorized into 10 groups with ?29% of the sequences having no matches in the databases. Immune related transcripts in the library were carboxypeptidase B, ferritin, translationally controlled tumor

S. Nayak; K. M. Ajay; N. Ramaiah; Ram M. Meena; R. A. Sreepada

2011-01-01

58

Assimilable Organic Carbon (AOC) in Soil Water Extracts Using Vibrio harveyi BB721 and Its Implication for Microbial Biomass  

Microsoft Academic Search

Assimilable organic carbon (AOC) is commonly used to measure the growth potential of microorganisms in water, but has not yet been investigated for measuring microbial growth potential in soils. In this study, a simple, rapid, and non-growth based assay to determine AOC in soil was developed using a naturally occurring luminous strain Vibrio harveyi BB721 to determine the fraction of

Jincai Ma; A. Mark Ibekwe; Menu Leddy; Ching-Hong Yang; David E. Crowley

2012-01-01

59

Evaluation of RAPD-PCR and protein profile analysis to differentiate Vibrio harveyi strains prevalent along the southwest coast of India  

Microsoft Academic Search

Sixty five isolates of Vibrio harveyi were subjected to random amplified polymorphic DNA (RAPD)-PCR analysis and protein profiling to investigate the genetic variability\\u000a among V. harveyi prevalent along the coast and also assess the discriminating ability of these two molecular methods. A total of 10 RAPD primers\\u000a were assayed for their specificity in detecting V. harveyi, of which only two

Biswajit Maiti; Malathi Shekar; Rekha Khushiramani; Iddya Karunasagar; Indrani Karunasagar

2009-01-01

60

Interference with the quorum sensing systems in a Vibrio harveyi strain alters the growth rate of gnotobiotically cultured rotifer Brachionus plicatilis  

Microsoft Academic Search

Aims: To evaluate the effect of Vibrio harveyi strains on the growth rate of the gnotobiotically cultured rotifer Brachionus plicatilis, and to establish whether quorum sensing is involved in the observed phenomena. Methods and Results: Gnotobiotic B. plicatilis sensu strictu, obtained by hatch- ing glutaraldehyde-treated amictic eggs, were used as test organisms. Challenge tests were performed with 11 V. harveyi

N. T. N. Tinh; N. D. Linh; T. K. Wood; K. Dierckens; P. Sorgeloos; P. Bossier

2007-01-01

61

The luxR gene product of Vibrio harveyi is a transcriptional activator of the lux promoter.  

PubMed Central

Expression of the lux operon from the marine bacterium Vibrio harveyi is dependent on cell density and requires an unlinked regulatory gene, luxR, and other cofactors for autoregulation. Escherichia coli transformed with the lux operon emits very low levels of light, and this deficiency can be partially alleviated by coexpression of luxR in trans. The V. harveyi lux promoter was analyzed in vivo by primer extension mapping to examine the function of luxR. RNA isolated from E. coli transformed with the Vibrio harveyi lux operon was shown to have a start site at 123 bp upstream of the first ATG codon of luxC. This is in sharp contrast to the start site found for lux RNA isolated from V. harveyi, at 26 bp upstream of the luxC initiation codon. However, when E. coli was cotransformed with both the lux operon and luxR, the start site of the lux mRNA shifted from -123 to -26. Furthermore, expression of the luxR gene caused a 350-fold increase in lux mRNA levels. The results suggest that LuxR of V. harveyi is a transcriptional activator stimulating initiation at the -26 lux promoter. Images

Swartzman, E; Silverman, M; Meighen, E A

1992-01-01

62

Multiplex PCR assay for identification of three major pathogenic Vibrio spp., Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus.  

PubMed

A multiplex PCR assay was developed based on atpA-sequence diversification for molecular identification of 3 major pathogenic Vibrio species: Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus. It specifically identified them from among 133 strains of various Vibrio species and other genera, and was applicable for testing seawater, suggesting its usefulness. PMID:21530641

Izumiya, Hidemasa; Matsumoto, Kazutoshi; Yahiro, Shunsuke; Lee, Jiyoung; Morita, Masatomo; Yamamoto, Shouji; Arakawa, Eiji; Ohnishi, Makoto

2011-08-01

63

Multiplex PCR assay for identification of three major pathogenic Vibrio spp., Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus  

Microsoft Academic Search

A multiplex PCR assay was developed based on atpA-sequence diversification for molecular identification of 3 major pathogenic Vibrio species: Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus. It specifically identified them from among 133 strains of various Vibrio species and other genera, and was applicable for testing seawater, suggesting its usefulness.

Hidemasa Izumiya; Kazutoshi Matsumoto; Shunsuke Yahiro; Jiyoung Lee; Masatomo Morita; Shouji Yamamoto; Eiji Arakawa; Makoto Ohnishi

2011-01-01

64

Analysing traces of autoinducer-2 requires standardization of the Vibrio harveyi bioassay.  

PubMed

Autoinducer-2 (furanosyl borate diester) is a biologically active compound whose role as a universal bacterial signalling molecule is currently under intense investigation. Because of its instability and the low concentrations of it found in biological samples, its detection relies at present on a bioassay that measures the difference in the timing of the luminescence of the Vibrio harveyi BB170 sensor strain with and without externally added AI-2. Here we systematically investigated which parameters affected the fold induction values of luminescence obtained in the bioassay and developed a modified protocol. Our experiments showed that growth and luminescence of V. harveyi BB170 are strongly influenced by trace elements. In particular, addition of Fe(3+) within a certain concentration range to the growth medium of the preinoculum culture improved the reproducibility and reduced the variance of the bioassay. In contrast, trace elements and vitamins introduced directly into the bioassay caused inhibitory effects. The initial density and luminescence of the sensor strain are very important and the values required for these parameters were defined. Borate interferes with the detection of AI-2 by giving false positive results. The response of V. harveyi BB170 to chemically synthesized AI-2 in the bioassay is nonlinear except over a very small concentration range; it is maximum over three orders of magnitude and shows inhibition above 35 microM. Based on the modified protocol, we were able to detect AI-2 in the absence of inhibitors with maximum fold induction values for the positive control (chemically synthesized AI-2) of >120 with a standard deviation of approximately 30% in a reliable and reproducible way. PMID:17143597

Vilchez, Ramiro; Lemme, André; Thiel, Verena; Schulz, Stefan; Sztajer, Helena; Wagner-Döbler, Irene

2007-01-01

65

A Nonluminescent and Highly Virulent Vibrio harveyi Strain Is Associated with "Bacterial White Tail Disease" of Litopenaeus vannamei Shrimp  

PubMed Central

Recurrent outbreaks of a disease in pond-cultured juvenile and subadult Litopenaeus vannamei shrimp in several districts in China remain an important problem in recent years. The disease was characterized by “white tail” and generally accompanied by mass mortalities. Based on data from the microscopical analyses, PCR detection and 16S rRNA sequencing, a new Vibrio harveyi strain (designated as strain HLB0905) was identified as the etiologic pathogen. The bacterial isolation and challenge tests demonstrated that the HLB0905 strain was nonluminescent but highly virulent. It could cause mass mortality in affected shrimp during a short time period with a low dose of infection. Meanwhile, the histopathological and electron microscopical analysis both showed that the HLB0905 strain could cause severe fiber cell damages and striated muscle necrosis by accumulating in the tail muscle of L. vannamei shrimp, which led the affected shrimp to exhibit white or opaque lesions in the tail. The typical sign was closely similar to that caused by infectious myonecrosis (IMN), white tail disease (WTD) or penaeid white tail disease (PWTD). To differentiate from such diseases as with a sign of “white tail” but of non-bacterial origin, the present disease was named as “bacterial white tail disease (BWTD)”. Present study revealed that, just like IMN and WTD, BWTD could also cause mass mortalities in pond-cultured shrimp. These results suggested that some bacterial strains are changing themselves from secondary to primary pathogens by enhancing their virulence in current shrimp aquaculture system.

Zhou, Junfang; Fang, Wenhong; Yang, Xianle; Zhou, Shuai; Hu, Linlin; Li, Xincang; Qi, Xinyong; Su, Hang; Xie, Layue

2012-01-01

66

Purification and characterization of fatty acyl-acyl carrier protein synthetase from Vibrio harveyi.  

PubMed Central

A Vibrio harveyi enzyme which catalyzes the ATP-dependent ligation of fatty acids to acyl carrier protein (ACP) has been purified 6,000-fold to apparent homogeneity by anion-exchange, gel filtration, and ACP-Sepharose affinity chromatography. Purified acyl-ACP synthetase migrated as a single 62-kDa band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and as an 80-kDa protein by gel filtration under reducing conditions. Activity of the purified enzyme was lost within hours in the absence of glycerol and low concentrations of Triton X-100. Acyl-ACP synthetase exhibited Kms for myristic acid, ACP, and ATP of 7 microM, 18 microM, and 0.3 mM, respectively. The enzyme was specific for adenine-containing nucleotides, and AMP was the product of the reaction. No covalent acyl-enzyme intermediate was observed. Enzyme activity was stimulated up to 50% by iodoacetamide but inhibited > 80% by N-ethylmaleimide: inhibition by the latter was prevented by ATP and ACP but not myristic acid. Dithiothreitol and sulfhydryl-directed reagents also influenced enzyme size, activity, and elution pattern on anion-exchange resins. The function of acyl-ACP synthetase has not been established, but it may be related to the capacity of V. harveyi to elongate exogenous fatty acids by an ACP-dependent mechanism. Images

Fice, D; Shen, Z; Byers, D M

1993-01-01

67

Cloning of the Vibrio harveyi luciferase genes: use of a synthetic oligonucleotide probe.  

PubMed Central

A mixed-sequence synthetic oligonucleotide probe was used to isolate a clone containing the gene encoding the alpha subunit of bacterial luciferase from Vibrio harveyi and part of the gene coding for the beta subunit. DNA sequence analysis has allowed us to determine that the genes are closely linked on the bacterial chromosome and transcribed in the same direction. Comparison of the sequences in the regions preceding the two structural genes has revealed considerable homology and has identified sites that may be involved in the expression of the genes. Identification of a clone from a clone bank of total genomic DNA from this organism shows that mixed probes can be successfully used to isolate a gene of interest from any bacterium provided some protein sequence for the gene product is available. Images

Cohn, D H; Ogden, R C; Abelson, J N; Baldwin, T O; Nealson, K H; Simon, M I; Mileham, A J

1983-01-01

68

Dynamics and Mechanism of A Quorum Sensing Network Regulated by Small RNAs in Vibrio Harveyi  

NASA Astrophysics Data System (ADS)

Bacterial quorum sensing (QS) has attracted much interests and it is an important process of cell communication. Recently, Bassler et al. studied the phenomena of QS regulated by small RNAs and the experimental data showed that small RNAs played important role in the QS of Vibrio harveyi and it can permit the fine-tuning of gene regulation and maintenance of homeostasis. According to Michaelis—Menten kinetics and mass action law in this paper, we construct a mathematical model to investigate the mechanism induced QS by coexist of small RNA and signal molecular (AI) and show that there are periodic oscillation when the time delay and Hill coefficient exceed a critical value and the periodic oscillation produces the change of concentration and induces QS. These results are fit to the experimental results. In the meanwhile, we also get some theoretical value of Hopf Bifurcation on time deday. In addition, we also find this network is robust against noise.

Shen, Jian-Wei

2011-03-01

69

Proteomic analysis of protein expression in the induction of the viable but Nonculturable State of Vibrio harveyi SF1.  

PubMed

Vibrio harveyi has been reported to enter into a viable but nonculturable (VBNC) state. One marine V. harveyi strain, SF1 became nonculturable when incubated in seawater microcosm at 4 °C within 60 days. We investigated protein expression in the exponential phase of V. harveyi SF1 and compared it to the VBNC state. Cytosolic proteins were resolved by two-dimensional polyacrylamide gel electrophoresis using pH 4-7 linear gradients. Among these proteins, sixteen proteins which were strongly downregulated or upregulated in the VBNC cells were identified by MALDI-TOF-TOF mass spectrometry. The results indicated that the differentially expressed proteins were mainly focused on stress response proteins and key components of central and intermediary metabolism, like carbohydrate metabolism, transport, and translation. This study provided clues for understanding the mechanism of adaptation to the VBNC state. PMID:23689940

Jia, Juntao; Li, Zhengyi; Cao, Jijuan; Jiang, Yinghui; Liang, Chengzhu; Liu, Mengzhen

2013-10-01

70

A Nitric Oxide-Responsive Quorum Sensing Circuit in Vibrio harveyi Regulates Flagella Production and Biofilm Formation  

PubMed Central

Cell signaling plays an important role in the survival of bacterial colonies. They use small molecules to coordinate gene expression in a cell density dependent manner. This process, known as quorum sensing, helps bacteria regulate diverse functions such as bioluminescence, biofilm formation and virulence. In Vibrio harveyi, a bioluminescent marine bacterium, four parallel quorum-sensing systems have been identified to regulate light production. We have previously reported that nitric oxide (NO), through the H-NOX/HqsK quorum sensing pathway contributes to light production in V. harveyi through the LuxU/LuxO/LuxR quorum sensing pathway. In this study, we show that nitric oxide (NO) also regulates flagellar production and enhances biofilm formation. Our data suggest that V. harveyi is capable of switching between lifestyles to be able to adapt to changes in the environment.

Henares, Bernadette M.; Xu, Yueming; Boon, Elizabeth M.

2013-01-01

71

Metabolomic analysis revealed the differential responses in two pedigrees of clam Ruditapes philippinarum towards Vibrio harveyi challenge.  

PubMed

Manila clam Ruditapes philippinarum is an important marine aquaculture shellfish. This species has several pedigrees including White, Zebra, Liangdao Red and Marine Red distributing in the coastal areas in North China. In this work, we studied the metabolic differences induced by Vibrio harveyi in hepatopancreas from White and Zebra clams using NMR-based metabolomics. Metabolic responses (e.g., amino acids, glucose, glycogen, ATP and succinate) and altered mRNA expression levels of related genes (ATP synthase, heat shock protein 90, defensin and lysozyme) suggested that V. harveyi induced clear disruption in energy metabolism and immune stresses in both White and Zebra clam hepatopancreas. However, V. harveyi caused obvious osmotic stress in Zebra clam hepatopancreas, which was not observed in V. harveyi-challenged White clams samples. In addition, V. harveyi challenge induced more severe disruption in energy metabolism and immune stress in White clams than in Zebra clams. Overall, our results indicated that the biological differences between different pedigrees of R. philippinarum should be considered in immunity studies. PMID:24161758

Liu, Xiaoli; Zhao, Jianmin; Wu, Huifeng; Wang, Qing

2013-12-01

72

Chromosomal replication origin from the marine bacterium Vibrio harveyi functions in Escherichia coli: oriC consensus sequence.  

PubMed Central

The chromosomal replication origin (oriC) of Vibrio harveyi has been isolated on a plasmid and shown to function as an origin in Escherichia coli. The nucleotide sequence of the V. harveyi oriC was determined. From a comparison of this sequence with oriC sequences of five enteric bacteria, we derived a consensus sequence of bacterial origins that function in E. coli. This consensus sequence identifies 122 positions within oriC where nucleotide substitutions can occur without loss of origin function. These positions are clustered rather than scattered. Four interrelated nine-base-pair repeats and eight of the dam methylation G-A-T-C sites are conserved in the consensus sequence. Very few relative insertion-deletion changes occur, and these are localized to one region of oriC. The genes for three polypeptides linked to the V. harveyi oriC were identified by using in vitro protein synthesis directed by deletion derivative plasmid templates. One of these genes, coding for a 58,000 Mr polypeptide and located 3.0 kilobase pairs from the V. harveyi oriC region, is lethal to E. coli when many copies (approximately 40 per cell) are present (high copy lethal or HCL gene). In addition, nucleotide sequence analysis showed that a different gene, the gid gene to the left of oriC, is highly conserved between E. coli and V. harveyi, whereas the coding region to the right of oriC is much less conserved. Images

Zyskind, J W; Cleary, J M; Brusilow, W S; Harding, N E; Smith, D W

1983-01-01

73

Use of the Bioluminescent Bacterium Vibrio harveyi to Detect Biohazardous Chemicals in Soil and Water Extractions with and without Acid  

Microsoft Academic Search

An investigation was undertaken using the bioluminescence-reduction bioassay of Vibrio harveyi to study the toxicity of 31 chemicals in a soil and water extraction that was treated with and without hydrochloric acid. Soil had the chemical added and was washed with 0.2 N HC1 and afterward this acid-treated soil containing the chemical was evaluated. Each chemical was tested independently. Endpoints

K. W. Thomulka; J. H. Lange

1995-01-01

74

Characterization of four lytic transducing bacteriophages of luminescent Vibrio harveyi isolated from shrimp (Penaeus monodon) hatcheries.  

PubMed

Four lytic bacteriophages designated as ?Vh1, ?Vh2, ?Vh3, and ?Vh4 were isolated from commercial shrimp hatcheries, possessing broad spectrum of infectivity against luminescent Vibrio harveyi isolates, considering their potential as biocontrol agent of luminescent bacterial disease in shrimp hatcheries, and were characterized by electron microscopy, genomic analysis, restriction enzyme analysis (REA), and pulsed-field gel electrophoresis (PFGE). Three phages ?Vh1, ?Vh2, and ?Vh4 had an icosahedral head of 60-115 nm size with a long, noncontractile tail of 130-329 × 1-17 nm, belonged to the family Siphoviridae. ?Vh3 had an icosahedral head (72 ± 5 nm) with a short tail (27 × 12 nm) and belonged to Podoviridae. REA with DraI and PFGE of genomic DNA digested with ScaI and XbaI and cluster analysis of their banding patterns indicated that ?Vh3 was distinct from the other three siphophages. PFGE-based genome mean size of the four bacteriophages ?Vh1, ?Vh2, ?Vh3, and ?Vh4 was estimated to be about 85, 58, 64, and 107 kb, respectively. These phages had the property of generalized transduction as demonstrated by transduction with plasmid pHSG 396 with frequencies ranging from 4.1 × 10(-7) to 2 × 10(-9) per plaque-forming unit, suggesting a potential ecological role in gene transfer among aquatic vibrios. PMID:22092866

Thiyagarajan, Sanjeevi; Chrisolite, Bagthasingh; Alavandi, Shankar V; Poornima, Modem; Kalaimani, Natarajan; Santiago, T Chinnappan

2011-12-01

75

MetR and CRP bind to the Vibrio harveyi lux promoters and regulate luminescence.  

PubMed

The induction of luminescence in Vibrio harveyi at the later stages of growth is controlled by a quorum-sensing mechanism in addition to nutritional signals. However, the mechanism of transmission of these signals directly to the lux promoters is unknown and only one regulatory protein, LuxR, has been shown to bind directly to lux promoter DNA. In this report, we have cloned and sequenced two genes, crp and metR, coding for the nutritional regulators, CRP (cAMP receptor protein) and MetR (a LysR homologue), involved in catabolite repression and methionine biosynthesis respectively. The metR gene was cloned based on a general strategy to detect lux DNA-binding proteins expressed from a genomic library, whereas the crp gene was cloned based on its complementation of an Escherichia coli crp mutant. Both CRP and MetR were shown to bind to lux promoter DNA, with CRP being dependent on the presence of cAMP. Expression studies indicated that the two regulators had opposite effects on luminescence: CRP was an activator and MetR a repressor. Disruption of crp decreased luminescence by about 1,000-fold showing that CRP is a major activator of luminescence the same as LuxR, whereas disruption of MetR resulted in activation of luminescence over 10-fold, confirming its function as a repressor. Comparison of the levels of the autoinducers involved in quorum sensing excreted by V. harveyi, and the crp and metR mutants, showed that autoinducer production was not significantly different, thus indicating that the nutritional signals do not affect luminescence by changing the levels of the signals required for quorum sensing. Indeed, the large effects of these nutritional sensors show that luminescence is controlled by multiple signals related to the environment and the cell density which must be integrated at the molecular level to control expression at the lux promoters. PMID:12366834

Chatterjee, Jaidip; Miyamoto, Carol M; Zouzoulas, Athina; Lang, B Franz; Skouris, Nicolas; Meighen, Edward A

2002-10-01

76

Biocontrol of Vibrio harveyi in Shrimp Aquaculture with Host Specific Lytic Bacteriophage  

Microsoft Academic Search

The current decline in shrimp farm production has been related to a variety of issues associated with survivability of shrimp grown in the aquaculture setting. The presence of Vibrio pathogens has been identified as a cause of high mortality in the post larval and juvenile shrimp in aquaculture. The use of bacteriophage to control infections has seen increased interest with

Lillian Barber

2012-01-01

77

Control of the Type 3 Secretion System in Vibrio harveyi by Quorum Sensing through Repression of ExsA ? ‡  

PubMed Central

The type 3 secretion system (T3SS) genes of Vibrio harveyi are activated at low cell density and repressed at high cell density by quorum sensing (QS). Repression requires LuxR, the master transcriptional regulator of QS-controlled genes. Here, we determine the mechanism underlying the LuxR repression of the T3SS system. Using a fluorescence-based cell sorting approach, we isolated V. harveyi mutants that are unable to express T3SS genes at low cell density and identified two mutations in the V. harveyi exsBA operon. While LuxR directly represses the expression of exsBA, complementation and epistasis analyses reveal that it is the repression of exsA expression, but not exsB expression, that is responsible for the QS-mediated repression of T3SS genes at high cell density. The present work further defines the genes in the V. harveyi QS regulon and elucidates a mechanism demonstrating how multiple regulators can be linked in series to direct the expression of QS target genes specifically at low or high cell density.

Waters, Christopher M.; Wu, Julie T.; Ramsey, Meghan E.; Harris, Rebecca C.; Bassler, Bonnie L.

2010-01-01

78

Control of the type 3 secretion system in Vibrio harveyi by quorum sensing through repression of ExsA.  

PubMed

The type 3 secretion system (T3SS) genes of Vibrio harveyi are activated at low cell density and repressed at high cell density by quorum sensing (QS). Repression requires LuxR, the master transcriptional regulator of QS-controlled genes. Here, we determine the mechanism underlying the LuxR repression of the T3SS system. Using a fluorescence-based cell sorting approach, we isolated V. harveyi mutants that are unable to express T3SS genes at low cell density and identified two mutations in the V. harveyi exsBA operon. While LuxR directly represses the expression of exsBA, complementation and epistasis analyses reveal that it is the repression of exsA expression, but not exsB expression, that is responsible for the QS-mediated repression of T3SS genes at high cell density. The present work further defines the genes in the V. harveyi QS regulon and elucidates a mechanism demonstrating how multiple regulators can be linked in series to direct the expression of QS target genes specifically at low or high cell density. PMID:20543047

Waters, Christopher M; Wu, Julie T; Ramsey, Meghan E; Harris, Rebecca C; Bassler, Bonnie L

2010-08-01

79

Biodesulfurization of dibenzothiophene in Escherichia coli is enhanced by expression of a Vibrio harveyi oxidoreductase gene  

SciTech Connect

One possible alternative to current fuel hydrodesulfurization methods is the use of microorganisms to remove sulfur compounds. Biodesulfurization requires much milder processing conditions, gives higher specificity, and does not require molecular hydrogen. In the present work the authors have produced two compatible plasmids: pDSR3, which allows Escherichia coli to convert dibenzothiophene (DBT) to hydroxybiphenyl (HBP), and pDSR2, which produces a Vibrio harveyi flavin oxidoreductase. The authors show that the flavin oxidoreductase enhances the rate of DBT removal when co-expressed in vivo with the desulfurization enzymes. The plasmids pDSR2 and pDSR3 were co-expressed in growing cultures. The expression of oxidoreductase caused an increase in the rate of DBT removal but a decrease in the rate of HBP production. The maximum rate of DBT removal was 8 mg/h {center{underscore}dot} g dry cell weight. Experiments were also conducted using resting cells with the addition of various carbon sources. It was found that the addition of glucose or glycerol to cultures with oxidoreductase expression produced the highest DBT removal rate. The culture with acetate and no oxidoreductase expression had the highest level of HBP production. For all carbon sources, the DBT removal rate was faster and the HBP generation rate slower with the expression of the oxidoreductase. Analysis of desulfurization intermediates indicates that the last enzyme in the pathway may be limiting.

Reichmuth, D.S.; Hittle, J.L.; Blanch, H.W.; Keasling, J.D.

2000-01-05

80

Biosynthesis and stereochemistry of the autoinducer controlling luminescence in Vibrio harveyi.  

PubMed Central

Knowledge of the pathway for synthesis of the autoinducer, N-(beta-hydroxybutyryl)-homoserine lactone (HBHL), controlling luminescence in Vibrio harveyi can provide important information concerning the relationship between the nutrition and physiology of the bacteria and the phenomenon of light emission. In this study, the D and L isomers of the autoinducer containing the stereoisomers of beta-hydroxybutyric acid were synthesized and characterized by proton nuclear magnetic resonance in the presence of a chiral shift reagent, a europium(III) derivative of Tris[3-(heptafluoropropyl-hydroxymethylene)-(+)-camphorato]. By using a newly isolated autoinducer mutant which responds to low physiological concentrations of the autoinducer, it could be shown that autoinducer activity was associated with D-HBHL and not L-HBHL. Blockage of fatty acid biosynthesis by the addition of fatty acids and/or the antibiotic cerulenin to the cells prevented synthesis of the autoinducer as measured by the loss of autoinducer activity and a decrease in the incorporation of labelled acetate into the partially purified autoinducer. These results indicate that fatty acid biosynthesis is necessary for light emission in luminescent bacteria because it controls formation of the lux autoinducer. Images

Cao, J G; Meighen, E A

1993-01-01

81

Crystallization and preliminary X-ray analysis of aldehyde dehydrogenase from Vibrio harveyi.  

PubMed Central

Aldehyde dehydrogenase from Vibrio harveyi catalyzes the oxidation of long-chain aliphatic aldehydes to acids. The enzyme is unique among the family of aldehyde dehydrogenases in that it exhibits much higher specificity for the cofactor NADP+ than for NAD+. The sequence of this form of the enzyme varies significantly from the NAD+ dependent forms, suggesting differences in the three-dimensional structure that may be correlated to cofactor specificity. Crystals of the enzyme have been grown both in the presence and absence of NADP+ using the hanging drop vapor diffusion technique. In order to improve crystal size and quality, iterative seeding techniques were employed. The crystals belong to space group P2(1), with unit cell dimensions a = 79.4 A, b = 131.1 A, c = 92.2 A, and beta = 92.4 degrees. Freezing the crystal to 100 K has enabled a complete set of data to be collected using a rotating anode source (lambda = 1.5418 A). The crystals diffract to a minimum d-spacing of 2.6 A resolution. Based on density calculations, two homodimers of molecular weight 110 kDa are estimated to be present in the asymmetric unit. Self-rotation functions show the presence of 3 noncrystallographic twofold symmetry axes.

Croteau, N.; Vedadi, M.; Delarge, M.; Meighen, E.; Abu-Abed, M.; Howell, P. L.; Vrielink, A.

1996-01-01

82

RNAi knock-down of the Litopenaeus vannamei Toll gene (LvToll) significantly increases mortality and reduces bacterial clearance after challenge with Vibrio harveyi.  

PubMed

In this study, we used real-time PCR to simultaneously monitor the responses of 12 key genes of the shrimp innate immune system in Litopenaeus vannamei after challenge with Vibrio harveyi. In the proPO activating system, we found that proPO was up-regulated (3.3x control at 36hpi). The hemolymph clotting genes transglutaminase (TGase) and clotting protein were also up-regulated, as were 5 genes in the antimicrobial peptide system (ALF, Crustin, Lyz, PEN2 and PEN4), with only PEN3 showing no significant changes. In the antioxidant defense system, SOD was slightly elevated while GPx was substantially down-regulated. In the pattern recognition receptor system, at 24hpi, the Toll gene (LvToll) showed the highest relative increase in expression level of all the investigated genes (15x greater than the sterile seawater control). In the second part of this study, when LvToll was knocked down by RNAi silencing, there was no effect on either survival rates or bacterial number in unchallenged shrimp. There was also no difference in mortality rates between control shrimp and LvToll-silenced shrimp when these two groups were challenged with a viral pathogen (white spot syndrome virus; WSSV). However, when LvToll-silenced shrimp were challenged by V. harveyi, there was a significant increase in mortality and bacterial CFU counts. We note that the increase in bacterial CFU count occurred even though treatment with EGFP dsRNA had the opposite effect of reducing the CFU counts. We conclude that LvToll is an important factor in the shrimp innate immune response to acute V. harveyi infection, but not to WSSV. PMID:19698743

Han-Ching Wang, Kc; Tseng, Chun-Wei; Lin, Han-You; Chen, I-Tung; Chen, Ya-Hui; Chen, Yi-Min; Chen, Tzong-Yueh; Yang, Huey-Lang

2010-01-01

83

In vivo and in vitro acylation of polypeptides in Vibrio harveyi: identification of proteins involved in aldehyde production for bioluminescence.  

PubMed Central

Incubation of soluble extracts from Vibrio harveyi with [3H]tetradecanoic acid (+ ATP) resulted in the acylation of several polypeptides, including proteins with molecular masses near 20 kilodaltons (kDa), and at least five polypeptides in the 30- to 60-kDa range. However, in growing cells pulse-labeled in vivo with [3H]tetradecanoic acid, only three of these polypeptides, with apparent molecular masses of 54, 42, and 32 kDa, were specifically labeled. When extracts were acylated with [3H] tetradecanoyl coenzyme A, on the other hand, only the 32-kDa polypeptide was labeled. When luciferase-containing dark mutants of V. harveyi were investigated, acylated 32-kDa polypeptide was not detected in a fatty acid-stimulated mutant, whereas the 42-kDa polypeptide appeared to be lacking in a mutant defective in aldehyde synthesis. Acylation of both of these polypeptides also increased specifically during induction of bioluminescence in V. harveyi. These results suggest that the role of the 32-kDa polypeptide is to supply free fatty acids, whereas the 42-kDa protein may be responsible for activation of fatty acids for their subsequent reduction to form the aldehyde substrates of the bioluminescent reaction. Images

Wall, L A; Byers, D M; Meighen, E A

1984-01-01

84

Synthesis and evaluation of thiazolidinedione and dioxazaborocane analogues as inhibitors of AI-2 quorum sensing in Vibrio harveyi.  

PubMed

Two focused libraries based on two types of compounds, that is, thiazolidinediones and dioxazaborocanes were designed. Structural resemblances can be found between thiazolidinediones and well-known furanone type quorum sensing (QS) inhibitors such as N-acylaminofuranones, and/or acyl-homoserine lactone signaling molecules, while dioxazaborocanes structurally resemble previously reported oxazaborolidine derivatives which antagonized autoinducer 2 (AI-2) binding to its receptor. Because of this, we hypothesized that these compounds could affect AI-2 QS in Vibrio harveyi. Although all compounds blocked QS, the thiazolidinediones were the most active AI-2 QS inhibitors, with EC(50) values in the low micromolar range. Their mechanism of inhibition was elucidated by measuring the effect on bioluminescence in a series of V. harveyi QS mutants and by DNA-binding assays with purified LuxR protein. The active compounds neither affected bioluminescence as such nor the production of AI-2. Instead, our results indicate that the thiazolidinediones blocked AI-2 QS in V. harveyi by decreasing the DNA-binding ability of LuxR. In addition, several dioxazaborocanes were found to block AI-2 QS by targeting LuxPQ. PMID:23286963

Brackman, Gilles; Al Quntar, Abed Al Aziz; Enk, Claes D; Karalic, Izet; Nelis, Hans J; Van Calenbergh, Serge; Srebnik, Morris; Coenye, Tom

2013-02-01

85

The Vibrio core group induces yellow band disease in Caribbean and Indo-Pacific reef-building corals  

Microsoft Academic Search

Aims: To determine the relationship between yellow band disease (YBD)- associated pathogenic bacteria found in both Caribbean and Indo-Pacific reefs, and the virulence of these pathogens. YBD is one of the most significant coral diseases of the tropics. Materials and Results: The consortium of four Vibrio species was isolated from YBD tissue on Indo-Pacific corals: Vibrio rotiferianus, Vibrio harveyi, Vibrio

J. M. Cervino; F. L. Thompson; B. Gomez-Gil; E. A. Lorence; T. J. Goreau; R. L. Hayes; K. B. Winiarski-Cervino; G. W. Smith; K. Hughen; E. Bartels

2008-01-01

86

Crystal structure of the NADP+-dependent aldehyde dehydrogenase from Vibrio harveyi: structural implications for cofactor specificity and affinity.  

PubMed Central

Aldehyde dehydrogenase from the bioluminescent bacterium, Vibrio harveyi, catalyses the oxidation of long-chain aliphatic aldehydes to acids. The enzyme is unique compared with other forms of aldehyde dehydrogenase in that it exhibits a very high specificity and affinity for the cofactor NADP(+). Structural studies of this enzyme and comparisons with other forms of aldehyde dehydrogenase provide the basis for understanding the molecular features that dictate these unique properties and will enhance our understanding of the mechanism of catalysis for this class of enzyme. The X-ray structure of aldehyde dehydrogenase from V. harveyi has been solved to 2.5-A resolution as a partial complex with the cofactor NADP(+) and to 2. 1-A resolution as a fully bound 'holo' complex. The cofactor preference exhibited by different forms of the enzyme is predominantly determined by the electrostatic environment surrounding the 2'-hydroxy or the 2'-phosphate groups of the adenosine ribose moiety of NAD(+) or NADP(+), respectively. In the NADP(+)-dependent structures the presence of a threonine and a lysine contribute to the cofactor specificity. In the V. harveyi enzyme an arginine residue (Arg-210) contributes to the high cofactor affinity through a pi stacking interaction with the adenine ring system of the cofactor. Further differences between the V. harveyi enzyme and other aldehyde dehydrogenases are seen in the active site, in particular a histidine residue which is structurally conserved with phosphorylating glyceraldehyde-3-phosphate dehydrogenase. This may suggest an alternative mechanism for activation of the reactive cysteine residue for nucleophilic attack.

Ahvazi, B; Coulombe, R; Delarge, M; Vedadi, M; Zhang, L; Meighen, E; Vrielink, A

2000-01-01

87

Exposure to Static Magnetic Field Stimulates Quorum Sensing Circuit in Luminescent Vibrio Strains of the Harveyi Clade  

PubMed Central

In this study, the evidence of electron-dense magnetic inclusions with polyhedral shape in the cytoplasm of Harveyi clade Vibrio strain PS1, a bioluminescent bacterium living in symbiosis with marine organisms, led us to investigate the behavior of this bacterium under exposure to static magnetic fields ranging between 20 and 2000 Gauss. When compared to sham-exposed, the light emission of magnetic field-exposed bacteria growing on solid medium at 18°C ±0.1°C was increased up to two-fold as a function of dose and growth phase. Stimulation of bioluminescence by magnetic field was more pronounced during the post-exponential growth and stationary phase, and was lost when bacteria were grown in the presence of the iron chelator deferoxamine, which caused disassembly of the magnetic inclusions suggesting their involvement in magnetic response. As in luminescent Vibrio spp. bioluminescence is regulated by quorum sensing, possible effects of magnetic field exposure on quorum sensing were investigated. Measurement of mRNA levels by reverse transcriptase real time-PCR demonstrated that luxR regulatory gene and luxCDABE operon coding for luciferase and fatty acid reductase complex were significantly up-regulated in magnetic field-exposed bacteria. In contrast, genes coding for a type III secretion system, whose expression was negatively affected by LuxR, were down-regulated. Up-regulation of luxR paralleled with down-regulation of small RNAs that mediate destabilization of luxR mRNA in quorum sensing signaling pathways. The results of experiments with the well-studied Vibrio campbellii strain BB120 (originally classified as Vibrio harveyi) and derivative mutants unable to synthesize autoinducers suggest that the effects of magnetic fields on quorum sensing may be mediated by AI-2, the interspecies quorum sensing signal molecule.

Tala, Adelfia; Delle Side, Domenico; Buccolieri, Giovanni; Tredici, Salvatore Maurizio; Velardi, Luciano; Paladini, Fabio; De Stefano, Mario; Nassisi, Vincenzo; Alifano, Pietro

2014-01-01

88

Nucleotide sequence of the LuxC gene and the upstream DNA from the bioluminescent system of Vibrio harveyi.  

PubMed Central

The nucleotide sequence of the luxC gene (1431 bp) and the upstream DNA (1049 bp) of the luminescent bacterium Vibrio harveyi has been determined. The luxC gene can be translated into a polypeptide of 55 kDa in excellent agreement with the molecular mass of the reductase polypeptide required for synthesis of the aldehyde substrate for the bioluminescent reaction. Analyses of codon usage showed a high frequency (1.9%) of the isoleucine codon, AUA, in the luxC gene compared to that found in Escherichia coli genes (0.2%) and its absence in the luxA, B and D genes. The low G/C content of the luxC gene and upstream DNA (38-39%) compared to that found in the other lux genes of V. harveyi (45%) was primarily due to a stretch of 500 nucleotides with only a 24% G/C content, extending from 200 bp inside lux C to 300 bp upstream. Moreover, an open reading frame did not extend for more than 48 codons between the luxC gene and 600 bp upstream at which point a gene transcribed in the opposite direction started. As the lux system in the luminescent bacterium, V. fischeri, contains a regulatory gene immediately upstream of luxC transcribed in the same direction, these results show that the organization and regulation of the lux genes have diverged in different luminescent bacteria.

Miyamoto, C M; Graham, A F; Meighen, E A

1988-01-01

89

Profiling of a few immune responsive genes expressed in postlarvae of Fenneropenaeus indicus challenged with Vibrio harveyi D3.  

PubMed

We identified 38 genes and eight hypothetical proteins by sequencing of 1200 clones from a Vibrio harveyi challenged Fenneropenaeus indicus subtracted cDNA library. Based on physiological roles and functions these genes were categorized into 10 groups with ?29% of the sequences having no matches in the databases. Immune related transcripts in the library were carboxypeptidase B, ferritin, translationally controlled tumor proteins (TCTP), hemocyanin, chitinase and serine carboxy peptidase. Remarkably, qPCR results imply 4.15, 3.45 and 1.86-fold rises in expression of ferritin, TCTP and hemocyanin transcripts respectively. Additionally, minor upregulation of other immune relevant genes lectin, penaeidin, crustin, MnSOD was observed in the challenged postlarvae. PMID:21515280

Nayak, S; Ajay, K M; Ramaiah, N; Meena, Ram M; Sreepada, R A

2011-06-01

90

Transcriptomic profiling of the oyster pathogen Vibrio splendidus opens a window on the evolutionary dynamics of the small RNA repertoire in the Vibrio genus  

PubMed Central

Work in recent years has led to the recognition of the importance of small regulatory RNAs (sRNAs) in bacterial regulation networks. New high-throughput sequencing technologies are paving the way to the exploration of an expanding sRNA world in nonmodel bacteria. In the Vibrio genus, compared to the enterobacteriaceae, still a limited number of sRNAs have been characterized, mostly in Vibrio cholerae, where they have been shown to be important for virulence, as well as in Vibrio harveyi. In addition, genome-wide approaches in V. cholerae have led to the discovery of hundreds of potential new sRNAs. Vibrio splendidus is an oyster pathogen that has been recently associated with massive mortality episodes in the French oyster growing industry. Here, we report the first RNA-seq study in a Vibrio outside of the V. cholerae species. We have uncovered hundreds of candidate regulatory RNAs, be it cis-regulatory elements, antisense RNAs, and trans-encoded sRNAs. Conservation studies showed the majority of them to be specific to V. splendidus. However, several novel sRNAs, previously unidentified, are also present in V. cholerae. Finally, we identified 28 trans sRNAs that are conserved in all the Vibrio genus species for which a complete genome sequence is available, possibly forming a Vibrio “sRNA core.”

Toffano-Nioche, Claire; Nguyen, An N.; Kuchly, Claire; Ott, Alban; Gautheret, Daniel; Bouloc, Philippe; Jacq, Annick

2012-01-01

91

Transcriptomic profiling of the oyster pathogen Vibrio splendidus opens a window on the evolutionary dynamics of the small RNA repertoire in the Vibrio genus.  

PubMed

Work in recent years has led to the recognition of the importance of small regulatory RNAs (sRNAs) in bacterial regulation networks. New high-throughput sequencing technologies are paving the way to the exploration of an expanding sRNA world in nonmodel bacteria. In the Vibrio genus, compared to the enterobacteriaceae, still a limited number of sRNAs have been characterized, mostly in Vibrio cholerae, where they have been shown to be important for virulence, as well as in Vibrio harveyi. In addition, genome-wide approaches in V. cholerae have led to the discovery of hundreds of potential new sRNAs. Vibrio splendidus is an oyster pathogen that has been recently associated with massive mortality episodes in the French oyster growing industry. Here, we report the first RNA-seq study in a Vibrio outside of the V. cholerae species. We have uncovered hundreds of candidate regulatory RNAs, be it cis-regulatory elements, antisense RNAs, and trans-encoded sRNAs. Conservation studies showed the majority of them to be specific to V. splendidus. However, several novel sRNAs, previously unidentified, are also present in V. cholerae. Finally, we identified 28 trans sRNAs that are conserved in all the Vibrio genus species for which a complete genome sequence is available, possibly forming a Vibrio "sRNA core." PMID:23097430

Toffano-Nioche, Claire; Nguyen, An N; Kuchly, Claire; Ott, Alban; Gautheret, Daniel; Bouloc, Philippe; Jacq, Annick

2012-12-01

92

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

PubMed

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. PMID:8186099

Aznar, R; Ludwig, W; Amann, R I; Schleifer, K H

1994-04-01

93

Vibrio fluvialis: an emerging human pathogen  

PubMed Central

Vibrio fluvialis is a pathogen commonly found in coastal environs. Considering recent increase in numbers of diarrheal outbreaks and sporadic extraintestinal cases, V. fluvialis has been considered as an emerging pathogen. Though this pathogen can be easily isolated by existing culture methods, its identification is still a challenging problem due to close phenotypic resemblance either with Vibrio cholerae or Aeromonas spp. However, using molecular tools, it is easy to identify V. fluvialis from clinical and different environmental samples. Many putative virulence factors have been reported, but its mechanisms of pathogenesis and survival fitness in the environment are yet to be explored. This chapter covers some of the major discoveries that have been made to understand the importance of V. fluvialis.

Ramamurthy, Thandavarayan; Chowdhury, Goutam; Pazhani, Gururaja P.; Shinoda, Sumio

2014-01-01

94

RNAi knock-down of the Litopenaeus vannamei Toll gene ( LvToll) significantly increases mortality and reduces bacterial clearance after challenge with Vibrio harveyi  

Microsoft Academic Search

In this study, we used real-time PCR to simultaneously monitor the responses of 12 key genes of the shrimp innate immune system in Litopenaeus vannamei after challenge with Vibrio harveyi. In the proPO activating system, we found that proPO was up-regulated (3.3× control at 36hpi). The hemolymph clotting genes transglutaminase (TGase) and clotting protein were also up-regulated, as were 5

KC Han-Ching Wang; Chun-Wei Tseng; Han-You Lin; I-Tung Chen; Ya-Hui Chen; Yi-Min Chen; Tzong-Yueh Chen; Huey-Lang Yang

2010-01-01

95

The lux autoinducer-receptor interaction in Vibrio harveyi: binding parameters and structural requirements for the autoinducer.  

PubMed Central

To assess the binding parameters and the structure-function relationship of the Vibrio harveyi lux autoinducer, N-(D-3-hydroxybutanoyl)homoserine lactone (D-HBHL), to light emission, a series of acylhomoserine lactone analogues were synthesized and their effects on the stimulation of luminescence of an autoinducer-deficient mutant of V. harveyi, D1, examined. Of the analogues with 3-hydroxyacyl chains, only N-(3-hydroxyvaleryl)homoserine lactone (HVHL) could act as an inducer, with about 85% of the potency of D-HBHL in stimulation of luminescence; the apparent Kd of the putative receptor for HVHL was 3.8 microM, close to that for the natural autoinducer (1.4 microM). Analogues with longer 3-hydroxyacyl chains, N-(3-hydroxyhexanoyl)homoserine lactone and N-(3-hydroxyheptanoyl)homoserine lactone, acted as competitive inhibitors of HBHL with apparent KI values of 77 and 53 microM respectively. Replacement of the 3-hydroxybutanoyl moiety with a 3-methylbutanoyl or 3-methoxybutanoyl group created weak competitive inhibitors, N-(isovaleryl)- and N-(3-methoxybutanoyl)- homoserine lactones, with apparent KI values of 150 and 360 microM respectively. Two other analogues, N-(2-hydroxybutanoyl)- and N-(4-hydroxybutanoyl)-homoserine lactone, could neither stimulate nor inhibit luminescence. The approach used in these studies to demonstrate binding of autoinducer analogues at the same site, as well as measurement of the relative dissociation constant, may be of value in analysing analogues activating or inhibiting luminescence and other processes that are under control of acylhomoserine lactone autoregulators.

Cao, J G; Wei, Z Y; Meighen, E A

1995-01-01

96

Studies on the immunomodulatory effect of polysaccharide gel extracted from Durio zibethinus in Penaeus monodon shrimp against Vibrio harveyi and WSSV.  

PubMed

Oral administration of polysaccharide gel (PG) in shrimp diets revealed immunostimulating potential and disease resistance in Penaeus monodon (black tiger shrimp). PG from the fruit-rind of Durio zibethinus has been characterized to be a pectic polysaccharide with immunomodulating and antibacterial activities. PG inhibited growth of the shrimp bacterial pathogen, Vibrio harveyi 1526, by agar diffusion and broth microdilution tests. Clear inhibition zones on agar plates were observed at the lowest PG concentration of 3.1 mg/ml, where minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values for PG were 6.3 and 12.5 mg/ml, respectively. Each group of juvenile shrimps, initial mean body weight 0.29 +/- 0.04 g, was housed in a closed-recirculating treated water system and was fed with PG-supplemented diets containing 1, 2 and 3% PG or shrimp basal diet in the control group for 8 and 12 weeks. PG-supplemented diets did not contribute to the overall growth of black tiger shrimp. The immune response was evaluated by analysis of prophenoloxidase activity and total hemocyte count in the shrimp fed PG-supplemented diets for 12 weeks. Prophenoloxidase activity in shrimp fed the 1, 2 and 3% PG-supplemented diet and total hemocyte count in shrimp fed the 1 and 2% PG-supplemented diet were higher (P < 0.05) than those of the control group. The percent survival was higher in groups fed the 1-3% PG-supplemented diets in challenge tests with either white spot syndrome virus (WSSV) or the bacterium V. harveyi 1526 than that of the control group. Relative percent survival (RPS) values in groups fed the 2% PG-supplemented diet showed the highest RPS value for disease resistance of 100% (at Day 6) and 36% (at Day 4) in treated shrimp against viral and bacterial infection, respectively. Mortality of PG-supplemented diets in treated shrimps against WSSV infection was also found to be much lower (P < 0.05) than that of the control group. PMID:20034573

Pholdaeng, Komsil; Pongsamart, Sunanta

2010-04-01

97

Redox-dependent sodium binding by the Na(+)-translocating NADH:quinone oxidoreductase from Vibrio harveyi.  

PubMed

Relaxation characteristics of the 23Na nuclei magnetization were used to determine the sodium-binding properties of the Na+-translocating NADH:quinone oxidoreductase from Vibrio harveyi (NQR). The dissociation constant of Na+ for the oxidized enzyme was found to be 24 mM and for the reduced enzyme about 30 microM. Such large (3 orders in magnitude) redox dependence of the NQR affinity to sodium ions shows that the molecular machinery was designed to use the drop in redox energy for creating an electrochemical sodium gradient. Redox titration of NQR monitored by changes in line width of the 23Na NMR signal at 2 mM Na+ showed that the enzyme affinity to sodium ions follows the Nernst law for a one-electron carrier with Em about -300 mV (vs SHE). The data indicate that energy conservation by NQR involves a mechanism modulating ion affinity by the redox state of an enzyme redox cofactor. PMID:17696408

Bogachev, Alexander V; Bertsova, Yulia V; Aitio, Olli; Permi, Perttu; Verkhovsky, Michael I

2007-09-01

98

Isolation of Vibrio harveyi acyl carrier protein and the fabG, acpP, and fabF genes involved in fatty acid biosynthesis.  

PubMed Central

We report the isolation of Vibrio harveyi acyl carrier protein (ACP) and cloning of a 3,973-bp region containing the fabG (encoding 3-ketoacyl-ACP reductase, 25.5 kDa), acpP (encoding ACP, 8.7 kDa), fabF (encoding 3-ketoacyl-ACP synthase II, 43.1 kDa), and pabC (encoding aminodeoxychorismate lyase, 29.9 kDa) genes. Predicted amino acid sequences were, respectively, 78, 86, 76, and 35% identical to those of the corresponding Escherichia coli proteins. Five of the 11 sequence differences between V. harveyi and E. coli ACP were nonconservative amino acid differences concentrated in a loop region between helices I and II.

Shen, Z; Byers, D M

1996-01-01

99

Prevention of quorum-sensing-mediated biofilm development and virulence factors production in Vibrio spp. by curcumin.  

PubMed

The increasing occurrence of disease outbreaks caused by Vibrio spp. and the emergence of antibiotic resistance has led to a growing interest in finding alternative strategies to prevent vibriosis. Since the pathogenicity of vibrios is controlled in part by quorum-sensing (QS) system, interfering with this mechanism would prevent the pathogenicity of vibrios without developing resistance. Hence, a non-toxic phytochemical curcumin from Curcuma longa was assessed for its potential in reducing the production of QS-dependent virulence factors in Vibrio spp. The obtained results evidenced 88% reduction in bioluminescence of Vibrio harveyi by curcumin. Further, curcumin exhibited a significant inhibition in alginate, exopolysaccharides, motility, biofilm development and other virulence factors production in Vibrio parahaemolyticus, Vibrio vulnificus and V. harveyi. In in vivo analysis, curcumin enhanced the survival rate of Artemia nauplii up to 67% against V. harveyi infection by attenuating its QS-mediated virulence. PMID:23354447

Packiavathy, Issac Abraham Sybiya Vasantha; Sasikumar, Pitchaikani; Pandian, Shunmugiah Karutha; Veera Ravi, Arumugam

2013-12-01

100

Inhibition of quorum sensing and biofilm formation in Vibrio harveyi by 4-fluoro-DPD; a novel potent inhibitor of signalling.  

PubMed

(S)-4,5-Dihydroxypentane-2,3-dione [(S)-DPD, (1)] is a precursor for , a quorum sensing signalling molecule for inter- and intra-species bacterial communication. The synthesis of its fluoro-analogue, 4-fluoro-5-hydroxypentane-2,3-dione () is reported. An intermediate in this route also enables a new, shorter synthesis of the native (S)-DPD. 4-Fluoro-DPD (2) completely inhibited bioluminescence and bacterial growth of Vibrio harveyi BB170 strain at 12.5 ?M and 100 ?M, respectively. PMID:24637781

Kadirvel, Manikandan; Fanimarvasti, Fariba; Forbes, Sarah; McBain, Andrew; Gardiner, John M; Brown, Gavin D; Freeman, Sally

2014-05-21

101

The anguibactin biosynthesis and transport genes are encoded in the chromosome of Vibrio harveyi: a possible evolutionary origin for the pJM1 plasmid-encoded system of Vibrio anguillarum?  

PubMed Central

Many Vibrio anguillarum serotype O1 strains carry 65-kb pJM1-type plasmids harboring genes involved in siderophore anguibactin biosynthesis and transport. The anguibactin system is an essential factor for V. anguillarum to survive under iron-limiting conditions, and as a consequence, it is a very important virulence factor of this bacterium. Our comparative analysis of genomic data identified a cluster harboring homologs of anguibactin biosynthesis and transport genes in the chromosome of Vibrio harveyi. We have purified the putative anguibactin siderophore and demonstrated that it is indeed anguibactin by mass spectrometry and specific bioassays. Furthermore, we characterized two genes, angR and fatA, in this chromosome cluster that, respectively, participate in anguibactin biosynthesis and transport as determined by mutagenesis analysis. Furthermore, we found that the V. harveyi FatA protein is located in the outer membrane fractions as previously demonstrated in V. anguillarum. Based on our data, we propose that the anguibactin biosynthesis and transport cluster in the V. anguillarum pJM1 plasmid have likely evolved from the chromosome cluster of V. harveyi or vice versa.

Naka, Hiroaki; Actis, Luis A; Crosa, Jorge H

2013-01-01

102

The soluble acyl-acyl carrier protein synthetase of Vibrio harveyi B392 is a member of the medium chain acyl-CoA synthetase family.  

PubMed

The gene encoding the unique soluble acyl-acyl carrier protein synthetase (AasS) of the bioluminescent Vibrio harveyi strain B392 has been isolated by expression cloning in Escherichia coli. This enzyme catalyzes the ATP-dependent acylation of the thiol of acyl carrier protein (ACP) with fatty acids with chain lengths from C4 to C18. The gene (called aasS) encodes a protein of 60 kDa, a hexahistidine-tagged version of which was readily expressed in E. coli and purified in large quantities. Surprisingly, the sequence of the encoded protein was significantly more similar to that of an acyl-CoA synthetase of the distantly related bacterium, Thermus thermophilus, than to that of the membrane-bound acyl-acyl carrier protein synthetase of E. coli, an enzyme that catalyzes the same reaction from a more closely related organism. Indeed, the AasS sequence can readily be modeled on the known crystal structures of the T. thermophilus acyl-CoA synthetase with remarkably high levels of conservation of the catalytic site residues. To test the possible role of AasS in the fatty aldehyde-dependent bioluminescence pathway of V. harveyi, the chromosomal aasS gene of the organism was disrupted by insertion of a kanamycin cassette by homologous recombination. The resulting aasS::kan strains retained low levels of acyl-acyl carrier protein synthetase consistent with prior indications of a second such activity in this bacterium. The mutant strains grew normally and had normal levels of bioluminescence but were deficient in the incorporation of exogenous octanoic acid into the cellular phospholipids of V. harveyi, particularly at low octanoate concentrations. These data indicate that AasS is responsible for a high-affinity and high-capacity uptake system that efficiently converts exogenous fatty acids into acyl-ACP species competent to enter the fatty acid biosynthetic cycle. PMID:16906759

Jiang, Yanfang; Chan, Chi Ho; Cronan, John E

2006-08-22

103

Severe Wound Infection with Photobacterium damselae ssp. damselae and Vibrio harveyi, following a Laceration Injury in Marine Environment: A Case Report and Review of the Literature  

PubMed Central

Marine microorganisms are uncommon etiologies of skin and skin structure infections, that is, wound infections. We report a case of severe wound infection, caused by the marine Photobacterium damselae (Vibrionaceae), in a 64-year-old male patient, returning from Australia. The isolate tested positive for pPHDD1, a plasmid conferring high-level virulence. Furthermore, the wound was coinfected with Vibrio harveyi, a halophile bacterium, which has never been reported from human infections before. Identification was achieved by use of Matrix-Assisted Laser Desorption-Ionization Time of Flight Mass Spectrometry (MALDI-TOF) and confirmed by 16S rDNA sequencing. Data retrieval from bibliography was complicated since P. damselae has been renamed often with a number of synonyms present in the literature: Photobacterium damsela, Vibrio damselae, Vibrio damsela, Pasteurella damselae, and Listonella damsela. With all synonyms used as query terms, a literature search provided less than 20 cases published worldwide. A majority of those cases presenting as severe wound infection are even fatal following progression into necrotizing fasciitis. Management with daily wound dressing and antibiotic therapy (ofloxacin empirically, followed by doxycycline after availability of microbiology) led in the reported case to a favorable outcome, which seems to be, however, the exception based on a review of the available literature.

Hundenborn, Jorg; Thurig, Steffi

2013-01-01

104

Identification of immune-related genes from kidney and spleen of turbot, Psetta maxima (L.), by suppression subtractive hybridization following challenge with Vibrio harveyi.  

PubMed

Suppression subtractive hybridization was used to investigate the response of turbot, Psetta maxima (L.), to Vibrio harveyi, by using a cDNA library constructed from artificially infected turbot kidney and spleen mRNA. Forty-nine expressed sequence tags were obtained. Several immune system genes were identified, including a major histocompatibility complex (MHC) class Ia gene and a heat shock protein 70 gene. Some signalling molecules were also present in the cDNA libraries, including src-family tyrosine kinase SCK, sgk-1 serine-threonine protein kinase and amyloid precursor-like protein 2. The full length of MHC class Ia cDNA was cloned from turbot cDNA by rapid amplification of cDNA ends polymerase chain reaction. The nucleotide sequence of turbot MHC class Ia has been submitted to GenBank with accession number EF032639. The turbot MHC class Ia cDNA has an open reading frame encoding 354 amino acids, and the deduced amino acid sequence of turbot MHC class Ia has 68%, 54%, 51%, 52%, 57%, 33%, 29% and 29% identities to those of olive flounder, medaka, rainbow trout, Atlantic cod, tiger puffer, chicken, mouse and human, respectively. Quantitative reverse transcriptase-PCR was performed for the MHC class Ia gene, and it was revealed that the expression level of the MHC class Ia gene in V. harveyi-challenged turbot increased to fourfold that of the controls. PMID:18577100

Wang, C; Zhang, X-H; Jia, A; Chen, J; Austin, B

2008-07-01

105

Quorum sensing regulation of virulence gene expression in Vibrio harveyi in vitro and in vivo during infection of gnotobiotic brine shrimp larvae.  

PubMed

In this study, we investigated quorum sensing regulation of virulence genes in Vibrio harveyi by determining their expression levels, both in vitro and in vivo during infection of gnotobiotic brine shrimp. The quorum sensing master regulator luxR and the vhp metalloprotease showed around threefold and fivefold higher expression levels in a luxO mutant with maximum quorum sensing activity than in a luxO mutant with minimum quorum sensing activity. There was no difference in expression of the vhh haemolysin gene between the two mutants. There was however more than 2.5-fold lower expression in an AI-2-negative mutant, suggesting that this gene is specifically regulated by AI-2 quorum sensing through a yet unknown signal transduction cascade. The in vivo expression data showed a peak in expression of the quorum sensing master regulator luxR and the vhp metalloprotease after 24?h of incubation in wild-type V. harveyi and the luxO mutant mimicking a maximally activated quorum sensing system, whereas the expression remained low in the luxO mutant mimicking a completely inactivated quorum sensing system. The vhh haemolysin gene showed a peak in expression after 24?h in the wild type and a constantly low expression in an AI-2-negative mutant. PMID:23761340

Ruwandeepika, H A Darshanee; Bhowmick, Patit Paban; Karunasagar, Indrani; Bossier, Peter; Defoirdt, Tom

2011-10-01

106

Vibriophages and Their Interactions with the Fish Pathogen Vibrio anguillarum.  

PubMed

Vibrio anguillarum is an important pathogen in aquaculture, responsible for the disease vibriosis in many fish and invertebrate species. Disease control by antibiotics is a concern due to potential development and spread of antibiotic resistance. The use of bacteriophages to control the pathogen may offer a non-antibiotic-based approach to reduce vibriosis. A detailed understanding of the phage-host interaction is needed to evaluate the potential of phages to control the pathogen. In this study, we examined the diversity and interactions of 11 vibriophages, 24 V. anguillarum strains, and 13 Vibrio species strains. Together, the host ranges of the 11 phages covered all of the tested 37 Vibrio sp. host strains, which represented considerable temporal (20 years) and geographical (9 countries) differences in their origins of isolation. Thus, despite the occurrence of unique susceptibility patterns of the individual host isolates, key phenotypic properties related to phage susceptibility are distributed worldwide and maintained in the global Vibrio community for decades. The phage susceptibility pattern of the isolates did not show any relation to the physiological relationships obtained from Biolog GN2 profiles, demonstrating that similar phage susceptibility patterns occur across broad phylogenetic and physiological differences in Vibrio strains. Subsequent culture experiments with two phages and two V. anguillarum hosts demonstrated an initial strong lytic potential of the phages. However, rapid regrowth of both phage-resistant and phage-sensitive cells following the initial lysis suggested that several mechanisms of protection against phage infection had developed in the host populations. PMID:24610858

Tan, Demeng; Gram, Lone; Middelboe, Mathias

2014-05-01

107

Analysis of Activator and Repressor Functions Reveals the Requirements for Transcriptional Control by LuxR, the Master Regulator of Quorum Sensing in Vibrio harveyi  

PubMed Central

ABSTRACT LuxR-type transcription factors are the master regulators of quorum sensing in vibrios. LuxR proteins are unique members of the TetR superfamily of transcription factors because they activate and repress large regulons of genes. Here, we used chromatin immunoprecipitation and nucleotide sequencing (ChIP-seq) to identify LuxR binding sites in the Vibrio harveyi genome. Bioinformatics analyses showed that the LuxR consensus binding site at repressed promoters is a symmetric palindrome, whereas at activated promoters it is asymmetric and contains only half of the palindrome. Using a genetic screen, we isolated LuxR mutants that separated activation and repression functions at representative promoters. These LuxR mutants exhibit sequence-specific DNA binding defects that restrict activation or repression activity to subsets of target promoters. Altering the LuxR DNA binding site sequence to one more closely resembling the ideal LuxR consensus motif can restore in vivo function to a LuxR mutant. This study provides a mechanistic understanding of how a single protein can recognize a variety of binding sites to differentially regulate gene expression.

van Kessel, Julia C.; Ulrich, Luke E.; Zhulin, Igor B.; Bassler, Bonnie L.

2013-01-01

108

Genome Sequence of the Pathogenic Bacterium Vibrio vulnificus Biotype 3  

PubMed Central

We report the first genome sequence of the pathogenic Vibrio vulnificus biotype 3. This draft genome sequence of the environmental strain VVyb1(BT3), isolated in Israel, provides a representation of this newly emerged clonal group, which reveals higher similarity to the clinical strains of biotype 1 than to the environmental ones.

Danin-Poleg, Yael; Elgavish, Sharona; Raz, Nili; Efimov, Vera

2013-01-01

109

Inhibition of Vibrio harveyi bioluminescence by cerulenin: In vivo evidence for covalent modification of the reductase enzyme involved in aldehyde synthesis  

SciTech Connect

Bacterial bioluminescence is very sensitive to cerulenin, a fungal antibiotic which is known to inhibit fatty acid synthesis. When Vibrio harveyi cells pretreated with cerulenin were incubated with ({sup 3}H)myristic acid in vivo, acylation of the 57-kilodalton reductase subunit of the luminescence-specific fatty acid reductase complex was specifically inhibited. Light emission of wild-type V. harveyi was 20-fold less sensitive to cerulenin at low concentrations (10{mu}g/ml) than that of the dark mutant strain M17, which requires exogenous myristic acid for luminescence because of a defective transferase subunit. The sensitivity of myristic acid-stimulated luminescence in the mutant strain M17 exceeded that of phospholipid synthesis from ({sup 14}C)acetate, whereas uptake and incorporation of exogenous ({sup 14}C)myristic acid into phospholipids was increased by cerulenin. The reductase subunit could be labeled by incubating M17 cells with ({sup 3}H)tetrahydrocerulenin; this labeling was prevented by preincubation with either unlabeled cerulenin or myristic acid. Labeling of the reductase subunit with ({sup 3}H)tetrahydrocerulenin was also noted in an aldehyde-stimulated mutant (A16) but not in wild-type cells or in another aldehyde-stimulated mutant (M42) in which ({sup 3}H)myristoyl turnover at the reductase subunit was found to be defective. These results indicate that (i) cerulenin specifically and covalently inhibits the reductase component of aldehyde synthesis, (ii) this enzyme is partially protected from cerulenin inhibition in the wild-type strain in vivo, and (iii) two dark mutants which exhibit similar luminescence phenotypes (mutants A16 and M42) are blocked at different stages of fatty acid reduction.

Byers, D.M. (Dalhousie Univ., Halifax (Nova Scotia)); Meighen, E.A. (McGill Univ., Montreal, Quebec (Canada))

1989-07-01

110

Inhibition of Vibrio harveyi bioluminescence by cerulenin: in vivo evidence for covalent modification of the reductase enzyme involved in aldehyde synthesis.  

PubMed Central

Bacterial bioluminescence is very sensitive to cerulenin, a fungal antibiotic which is known to inhibit fatty acid synthesis. When Vibrio harveyi cells pretreated with cerulenin were incubated with [3H]myristic acid in vivo, acylation of the 57-kilodalton reductase subunit of the luminescence-specific fatty acid reductase complex was specifically inhibited. In contrast, in vitro acylation of both the synthetase and transferase subunits, as well as the activities of luciferase, transferase, and aldehyde dehydrogenase, were not adversely affected by cerulenin. Light emission of wild-type V. harveyi was 20-fold less sensitive to cerulenin at low concentrations (10 micrograms/ml) than that of the dark mutant strain M17, which requires exogenous myristic acid for luminescence because of a defective transferase subunit. The sensitivity of myristic acid-stimulated luminescence in the mutant strain M17 exceeded that of phospholipid synthesis from [14C]acetate, whereas uptake and incorporation of exogenous [14C]myristic acid into phospholipids was increased by cerulenin. The reductase subunit could be labeled by incubating M17 cells with [3H]tetrahydrocerulenin; this labeling was prevented by preincubation with either unlabeled cerulenin or myristic acid. Labeling of the reductase subunit with [3H]tetrahydrocerulenin was also noted in an aldehyde-stimulated mutant (A16) but not in wild-type cells or in another aldehyde-stimulated mutant (M42) in which [3H]myristoyl turnover at the reductase subunit was found to be defective. These results indicate that (i) cerulenin specifically and covalently inhibits the reductase component of aldehyde synthesis, (ii) this enzyme is partially protected from cerulenin inhibition in the wild-type strain in vivo, and (iii) two dark mutants which exhibit similar luminescence phenotypes (mutants A16 and M42) are blocked at different stages of fatty acid reduction. Images

Byers, D M; Meighen, E A

1989-01-01

111

Vibrio jasicida sp. nov., a member of the Harveyi clade, isolated from marine animals (packhorse lobster, abalone and Atlantic salmon).  

PubMed

Six isolates of a facultatively anaerobic bacterium were recovered in culture from marine invertebrates and vertebrates, including packhorse lobster (Jasus verreauxi), abalone (Haliotis sp.) and Atlantic salmon (Salmo salar), between 1994 and 2002. The bacteria were Gram-negative, rod-shaped and motile by means of more than one polar flagellum, oxidase-positive, catalase-positive and able to grow in the presence of 0.5-8.0% NaCl (optimum 3.0-6.0%) and at 10-37 °C (optimum 25-30 °C). On the basis of 16S rRNA gene sequence analysis and multilocus sequence analysis (MLSA) using five loci (2443 bp; gyrB, pyrH, ftsZ, mreB and gapA), the closest phylogenetic neighbours of strain TCFB 0772(T) were the type strains of Vibrio communis (99.8 and 94.6?% similarity, respectively), Vibrio owensii (99.8 and 94.1%), Vibrio natriegens (99.4 and 88.8%), Vibrio parahaemolyticus (99.4 and 90.3%), Vibrio rotiferianus (99.2 and 94.4%), Vibrio alginolyticus (99.1 and 89.3%) and Vibrio campbellii (99.1 and 92.3%). DNA-DNA hybridization confirmed that the six isolates constitute a unique taxon that is distinct from other known species of Vibrio. In addition, this taxon can be readily differentiated phenotypically from other Vibrio species. The six isolates therefore represent a novel species, for which the name Vibrio jasicida sp. nov. is proposed; the novel species is represented by the type strain TCFB 0772(T) (?=?JCM 16453(T) ?=?LMG 25398(T)) (DNA G+C content 45.9 mol%) and reference strains TCFB 1977 (?=?JCM 16454) and TCFB 1000 (?=?JCM 16455). PMID:21984666

Yoshizawa, Susumu; Tsuruya, Yasuhiro; Fukui, Youhei; Sawabe, Tomoo; Yokota, Akira; Kogure, Kazuhiro; Higgins, Melissa; Carson, Jeremy; Thompson, Fabiano L

2012-08-01

112

Low Dose Gamma Irradiation to Reduce Pathogenic Vibrios in Live Oysters (Crassostrea virginica)  

Microsoft Academic Search

Pathogenic strains of Vibrio (Vibrio vulnificus and V. parahaemolyticus), natural inhabitants of estuarine and ocean environments, can cause serious illness and death in susceptible persons from consumption of raw half-shell oysters. Objectives of this study were (1) to establish the irradiation dose needed to reduce pathogenic vibrios to nondetectable levels and (2) to determine consumer's ability to differentiate between irradiated

Linda Andrews; Michael Jahncke; Kumar Mallikarjunan

2003-01-01

113

Benthic ecology of Vibrio spp. and pathogenic Vibrio species in a coastal Mediterranean environment (La Spezia Gulf, Italy).  

PubMed

We carried out a 16-month in situ study to investigate the ecology of Vibrio spp. and pathogenic Vibrio species in coastal sediments of the Mediterranean Sea, employing multiple-regression analysis to reveal the major environmental factors controlling their occurrence in the benthic environment. In addition, association between vibrios and sediment-inhabiting meiofauna, which is a major component of benthic ecosystems, was investigated. Culturable and total Vibrio spp. estimates by most-probable-number technique coupled with standard polymerase chain reaction (PCR) and real-time PCR methods, respectively, were at least one order of magnitude higher in sediment than in seawater. In addition, potential human pathogenic species Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus occurred in the sediment with V. parahaemolyticus being the most frequently found. In the pelagic environment, 60% of total variance in culturable Vibrio data was explained by sea surface temperature (40%), salinity (13%) and organic matter concentration (7%). In the benthic environment, sea surface temperature was the only factor that significantly affected culturable Vibrio occurrence although it explained only 25% of total variance, suggesting that additional unexplored factors may play a role as well. No correlation was found between culturable Vibrio spp. concentrations and the abundance of harpacticoid copepods in the sediment whilst a negative correlation was found between Vibrio spp. and nematode abundance which accounted for almost 90% of the total meiofaunal density. Taxonomic analysis revealed that selective bacterial feeders accounted for nearly 50% of the total nematode community and included genera such as Terschellingia, Molgolaimus and Halalaimus, suggesting that top-down control by nematode grazing may be an important factor affecting Vibrio occurrence in these sediments. It is concluded that the benthic marine environment may function as a reservoir of Vibrio spp. and potential pathogenic vibrios whose ecological features appeared substantially different from the ones recognised in the pelagic environment. PMID:19543938

Vezzulli, Luigi; Pezzati, Elisabetta; Moreno, Mariapaola; Fabiano, Mauro; Pane, Luigi; Pruzzo, Carla

2009-11-01

114

Expression of Vibrio harveyi Acyl-ACP Synthetase Allows Efficient Entry of Exogenous Fatty Acids into the Escherichia coli Fatty Acid and Lipid A Synthetic Pathways  

PubMed Central

Although the Escherichia coli fatty acid synthesis (FAS) pathway is the best studied type II fatty acid synthesis system, a major experimental limitation has been the inability to feed intermediates into the pathway in vivo because exogenously-supplied free fatty acids are not efficiently converted to the acyl-acyl carrier protein (ACP) thioesters required by the pathway. We report that expression of Vibrio harveyi acyl-ACP synthetase (AasS), a soluble cytosolic enzyme that ligates free fatty acids to ACP to form acyl-ACPs, allows exogenous fatty acids to enter the E. coli fatty acid synthesis pathway. The free fatty acids are incorporated intact and can be elongated or directly incorporated into complex lipids by acyltransferases specific for acyl-ACPs. Moreover, expression of AasS strains and supplementation with the appropriate fatty acid restored growth to E. coli mutant strains that lack essential fatty acid synthesis enzymes. Thus, this strategy provides a new tool for circumventing the loss of enzymes essential for FAS function.

Jiang, Yanfang; Morgan-Kiss, Rachael M.; Campbell, John W.; Chan, Chi Ho; Cronan, John E.

2010-01-01

115

Comparative Genome Analysis of Vibrio vulnificus, a Marine Pathogen  

Microsoft Academic Search

The halophile Vibrio vulnificus is an etiologic agent of human mortality from seafood-borne infections. We applied whole-genome sequencing and comparative analysis to investigate the evolution of this pathogen. The genome of biotype 1 strain, V. vulnificus YJ016, was sequenced and includes two chromosomes of estimated 3377 kbp and 1857 kbp in size, and a plasmid of 48,508 bp. A super-integron

Chung-Yung Chen; Keh-Ming Wu; Yo-Cheng Chang; Chuan-Hsiung Chang; Hui-Chi Tsai; Tsai-Lien Liao; Yen-Ming Liu; Hsiang-Ju Chen; Arthur Bo-Ting Shen; Jian-Chiuan Li; Teh-Li Su; Chung-Ping Shao; Chung-Te Lee; Lien-I Hor; Shih-Feng Tsai

2003-01-01

116

Vibrio ostreicida sp. nov., a new pathogen of bivalve larvae.  

PubMed

The taxonomic position of the bivalve pathogen PP-203(T) was studied together with those of two similar isolates (PP-200 and PP-204). The bacterial strains were isolated from samples of young oyster spat in a bivalve hatchery in Galicia (NW Spain), which was continually affected by outbreaks of disease and severe mortalities. On the basis of 16S rRNA gene sequencing, the three strains formed a cluster within the genus Vibrio and were most closely related to Vibrio pectenicida DSM 19585(T) (97.9?% similarity). Additional multilocus sequence analysis, including sequences of the housekeeping genes rpoA, recA, pyrH, gyrB and ftsZ, and DNA-DNA hybridization experiments indicated that the strains were distinct from currently known species of the genus Vibrio and confirmed the clustering of the three isolates. Several phenotypic features, such as growth in TCBS medium and nitrate reduction, proved useful for distinguishing the proposed novel species from its closest relatives. The findings support the description of a novel species to include the three isolates, for which the name Vibrio ostreicida sp. nov. (type strain PP-203(T)?=?CECT 7398(T)?=?DSM 21433(T)) is proposed. PMID:24510976

Prado, Susana; Dubert, Javier; Romalde, Jesús L; Toranzo, Alicia E; Barja, Juan L

2014-05-01

117

Biocontrol of pathogens in shrimp hatcheries using bacteriophages  

Microsoft Academic Search

Microflora associated with larval stages of shrimp could affect the health and development of the larvae. Some bacteria such as luminous Vibrio harveyi cause serious mortalities. Consequent to the ban on use of most antibiotics in aquaculture, there is a need for alternate technologies for control of bacterial pathogens. Bacteriophages have a potential to control bacterial pathogens. This paper reports

Indrani Karunasagar; M. M. Shivu; S. K. Girisha; G. Krohne; Iddya Karunasagar

2007-01-01

118

Structure-function relationship of Vibrio harveyi NADPH-flavin oxidoreductase FRP: essential residues Lys167 and Arg15 for NADPH binding.  

PubMed

Vibrio harveyi NADPH-FMN oxidoreductase (FRP) catalyzes flavin reduction by NADPH. In comparing amino acid sequence and crystal structure with Escherichia coli NfsA, residues N134, R225, R133, K167, and R15 were targeted for investigation of their possible roles in the binding and utilization of the NADPH substrate. By mutation of each of these five residues to an alanine, steady-state rate analyses showed that the variants K167A and R15A had apparently greatly increased K(m,NADPH) and reduced k(cat)/K(m,NADPH), whereas little or much more modest changes were found for the other variants. The deuterium isotope effects (D)(V/K) for (4R)-[4-(2)H]-NADPH were markedly increased to 6.3 and 7.4 for K167A and R15A, respectively, indicating that the rate constants for NADPH and NADP(+) dissociation were greatly enhanced relative to the hydride transfer steps. Also, anaerobic stopped-flow analyses revealed that the equilibrium dissociation constant for NADPH binding (K(d)) to be 2.5-3.9 and 1.1 mM for K167A and R15A, respectively, much higher than the 0.4 ?M K(d) for the native FRP, whereas the k(cat) of these two variants were similar to that of the wild-type enzyme. Moreover, the K167 to alanine mutation led to even a slight increase in k(cat)/K(m) for NADH. These results, taken together, provide a strong support to the conclusion that K167 and R15 each was critical in the binding of NADPH by FRP. Such a functional role may also exist for other FRP homologous proteins. PMID:22650604

Chung, Hae-Won; Tu, Shiao-Chun

2012-06-19

119

Design and validation of a novel multiplex real-time PCR assay for Vibrio pathogen detection.  

PubMed

Three species--Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus--account for the majority of vibrio infections in humans. Rapid and accurate identification of Vibrio species has been problematic because phenotypic characteristics are variable within species. Additionally, biochemical identification and confirmation require 2 or more days to complete. Rapid and sensitive molecular techniques for the detection of vibrio pathogens would be useful for the surveillance and management of outbreaks. To facilitate the identification of human-pathogenic species, we designed and validated a highly sensitive, specific, and robust multiplex real-time PCR assay to identify V. cholerae, V. parahaemolyticus, and V. vulnificus using a four-dye configuration in a convenient lyophilized format. Multiple Vibrio strains were sequenced to verify candidate target TaqMan sites. Several individual assays within the multiplex contain multiple primers or probes to ensure detection of polymorphic variants. V. cholerae, V. parahaemolyticus, and V. vulnificus were detected either individually or in mixtures at ?30 genomic copies. V. cholerae was specifically detected in the presence or absence of Vibrio mimicus. The Vibrio multiplex assay showed 100% specificity to all targets analyzed and no detection of nearest neighbor strains. Each assay exhibited 100% ± 10% efficiency. Multiplex real-time PCR can simplify pathogen detection and reduce costs per test since three species can be analyzed in a single reaction tube. Rapid and accurate detection of pathogenic vibrios in shellfish or seawater samples will improve the microbiological safety of seafood for consumers. PMID:21669071

Tebbs, Robert S; Brzoska, Pius M; Furtado, Manohar R; Petrauskene, Olga V

2011-06-01

120

Phage therapy treatment of the coral pathogen Vibrio coralliilyticus  

PubMed Central

Vibrio coralliilyticus is an important coral pathogen demonstrated to cause disease outbreaks worldwide. This study investigated the feasibility of applying bacteriophage therapy to treat the coral pathogen V. coralliilyticus. A specific bacteriophage for V. coralliilyticus strain P1 (LMG23696), referred to here as bacteriophage YC, was isolated from the seawater above corals at Nelly Bay, Magnetic Island, central Great Barrier Reef (GBR), the same location where the bacterium was first isolated. Bacteriophage YC was shown to be a lytic phage belonging to the Myoviridae family, with a rapid replication rate, high burst size, and high affinity to its host. By infecting its host bacterium, bacteriophage YC was able to prevent bacterial-induced photosystem inhibition in pure cultures of Symbiodinium, the photosymbiont partner of coral and a target for virulence factors produced by the bacterial pathogen. Phage therapy experiments using coral juveniles in microtiter plates as a model system revealed that bacteriophage YC was able to prevent V. coralliilyticus-induced photoinactivation and tissue lysis. These results demonstrate that bacteriophage YC has the potential to treat coral disease outbreaks caused by the bacterial pathogen V. coralliilyticus, making it a good candidate for phage therapy treatment of coral disease.

Cohen, Yossi; Joseph Pollock, F; Rosenberg, Eugene; Bourne, David G

2013-01-01

121

Interactions between Mytilus galloprovincialis hemocytes and the bivalve pathogens Vibrio aestuarianus 01/032 and Vibrio splendidus LGP32.  

PubMed

Marine bivalves can accumulate large numbers of bacteria, in particular Vibrio species, whose persistence in bivalve tissues largely depends on their sensitivity to the bactericidal activity of circulating hemocytes and hemolymph soluble factors. The interactions between vibrios and hemolymph have been investigated, in particular in bivalve species susceptible to infection by certain Vibrio spp. and strains. In this work, the effects of two bivalve pathogens, Vibrio splendidus LGP32 (V.s.) and Vibrio aestuarianus 01/032 (V.a.), isolated from oyster mortality outbreaks, on the hemocytes of Mytilus galloprovincialis were investigated. In vitro, V.s., but not V.a., induced a dramatic decrease in lysosomal membrane stability-LMS in the hemocytes; both vibrios induced a moderate lysozyme release, with V.s. > V.a.. The V.s.-induced decrease in LMS was mediated by activation of PI-3Kinase, as shown by use of different kinase inhibitors. TEM analysis showed rapid internalization of both vibrios; however, V.s. lead to cellular and lysosomal damage and was able to survive within the hemocytes, whereas significant killing of V.a. was observed. In vivo, in mussels challenged with either vibrio and sampled at 6, 24 and 96 h post-injection, transient decreases in hemocyte LMS and progressive increases in serum lysozyme activity were observed, with V.s. > V.a.. Moreover, whereas V.a. was efficiently cleared from hemolymph, V.s. showed significant growth, that was maximal at 24 h p.i. when lowest LMS values were recorded in the hemocytes. Both vibrios also induced significant decreases in LMS in the digestive gland, again with V.s. > V.a.. The results indicate distinct interactions between mussel hemocytes and the two vibrio strains tested. The effects of V.s. may be due to the capacity of this strain to interfere with the signaling pathways involved in hemocyte function, thus escaping the bactericidal activity of the host cell, as observed for certain mammalian pathogens. Although V.s. is considered not pathogenic to Mytilus, this vibrio strain can affect the lysosomal function at the cellular and tissue level, thus leading to stressful conditions. PMID:24080469

Balbi, T; Fabbri, R; Cortese, K; Smerilli, A; Ciacci, C; Grande, C; Vezzulli, L; Pruzzo, C; Canesi, L

2013-12-01

122

Pathogen Special: Vibrio Cholerae, Pseudomonas Aeruginosa and Xylella Fastidiosa  

PubMed Central

One could almost say that it is the latest fashion to sequence a bacterial genome. However, this would belittle the efforts of those working on these important organisms, whose data will greatly help those working on the prevention of disease in the fields of medicine and agriculture. In this feature we present a guided tour of the latest additions to the ‘sequenced microbes’ club. Vibrio cholerae is the causative agent of cholera, which is still a threat in countries with poor sanitation and unsafe drinking water. Pseudomonas aeruginosa is responsible for a large proportion of opportunistic human infections, typically infecting those with compromised immune systems, particularly cystic fibrosis patients, those patients on respirators and burn victims. Xylella fastidiosa is a plant pathogen that attacks citrus fruits by blocking the xylem, resulting in juiceless fruits of no commercial value.

2000-01-01

123

The human pathogenic vibrios--a public health update with environmental perspectives.  

PubMed Central

Pathogenic Vibrio species are naturally-occurring bacteria in freshwater and saline aquatic environments. Counts of free-living bacteria in water are generally less than required to induce disease. Increases in number of organisms towards an infective dose can occur as water temperatures rise seasonally followed by growth and concentration of bacteria on higher animals, such as chitinous plankton, or accumulation by shellfish and seafood. Pathogenic Vibrio species must elaborate a series of virulence factors to elicit disease in humans. Activities which predispose diarrhoeal and extraintestinal infections include ingestion of seafood and shellfish and occupational or recreational exposure to natural aquatic environments, especially those above 20 degrees C. Travel to areas endemic for diseases due to pathogenic Vibrio species may be associated with infections. Host risk factors strongly associated with infections are lack of gastric acid and liver disorders. Involvement of pathogenic Vibrio species in cases of diarrhoea should be suspected especially if infection is associated with ingestion of seafood or shellfish, raw or undercooked, in the previous 72 h. Vibrio species should be suspected in any acute infection associated with wounds sustained or exposed in the marine or estuarine environment. Laboratories serving coastal areas where infection due to pathogenic Vibrio species are most likely to occur should consider routine use of TCBS agar and other detection regimens for culture of Vibrio species from faeces, blood and samples from wound and ear infections.

West, P. A.

1989-01-01

124

Involvement of LuxR, a quorum sensing regulator in Vibrio harveyi, in the promotion of metabolic genes: argA, purM, lysE and rluA.  

PubMed

Quorum sensing, involving signal transduction via the two-component response regulator LuxO to its downstream target LuxR, controls luminescence in the marine bacterium Vibrio harveyi. LuxR is a DNA binding protein that acts as both activator of the lux operon and repressor of its own gene. In order to determine if any other genes are affected by quorum sensing in V. harveyi, an assay for luxR-dependent promotion was devised using a genomic library maintained in a novel luxAB (luciferase) reporter. Screening in Escherichia coli DH-21 (lacI(sq)) entailed the addition of a second plasmid containing luxR under plac control. Four out of 5000 colonies showed luminescence stimulation upon IPTG induction of luxR. The four luxR-dependent promoters were upstream of argA, purM, lysE, and rluA, genes involved in arginine and purine biosyntheses, amino acid efflux, and pseudouridine synthesis, respectively. Based on analysis of luxR-dependent promoters, particularly that of argA, we describe a LuxR binding site, and implicate the coordination of LuxR with ArgR. PMID:16844243

Miyamoto, Carol M; Meighen, Edward A

2006-06-01

125

Experimental Reservoirs of Human Pathogens: The Vibrio Cholerae Paradigm (7th Annual SFAF Meeting, 2012)  

ScienceCinema

Rita Colwell on "Experimental Reservoirs of Human Pathogens: The Vibrio cholerae paradigm" at the 2012 Sequencing, Finishing, Analysis in the Future Meeting held June 5-7, 2012 in Santa Fe, New Mexico.

Colwell, Rita [University of Maryland

2013-02-12

126

Persistence, seasonal dynamics and pathogenic potential of Vibrio communities from Pacific oyster hemolymph.  

PubMed

Bacteria of the genus Vibrio occur at a continuum from free-living to symbiotic life forms, including opportunists and pathogens, that can contribute to severe diseases, for instance summer mortality events of Pacific oysters Crassostrea gigas. While most studies focused on Vibrio isolated from moribund oysters during mortality outbreaks, investigations of the Vibrio community in healthy oysters are rare. Therefore, we characterized the persistence, diversity, seasonal dynamics, and pathogenicity of the Vibrio community isolated from healthy Pacific oysters. In a reciprocal transplant experiment we repeatedly sampled hemolymph from adult Pacific oysters to differentiate population from site-specific effects during six months of in situ incubation in the field. We characterized virulence phenotypes and genomic diversity based on multilocus sequence typing in a total of 70 Vibrio strains. Based on controlled infection experiments we could show that strains with the ability to colonize healthy adult oysters can also have the potential to induce high mortality rates on larvae. Diversity and abundance of Vibrio varied significantly over time with highest values during and after spawning season. Vibrio communities from transplanted and stationary oysters converged over time, indicating that communities were not population specific, but rather assemble from the surrounding environment forming communities, some of which can persist over longer periods. PMID:24728233

Wendling, Carolin C; Batista, Frederico M; Wegner, K Mathias

2014-01-01

127

Persistence, Seasonal Dynamics and Pathogenic Potential of Vibrio Communities from Pacific Oyster Hemolymph  

PubMed Central

Bacteria of the genus Vibrio occur at a continuum from free-living to symbiotic life forms, including opportunists and pathogens, that can contribute to severe diseases, for instance summer mortality events of Pacific oysters Crassostrea gigas. While most studies focused on Vibrio isolated from moribund oysters during mortality outbreaks, investigations of the Vibrio community in healthy oysters are rare. Therefore, we characterized the persistence, diversity, seasonal dynamics, and pathogenicity of the Vibrio community isolated from healthy Pacific oysters. In a reciprocal transplant experiment we repeatedly sampled hemolymph from adult Pacific oysters to differentiate population from site-specific effects during six months of in situ incubation in the field. We characterized virulence phenotypes and genomic diversity based on multilocus sequence typing in a total of 70 Vibrio strains. Based on controlled infection experiments we could show that strains with the ability to colonize healthy adult oysters can also have the potential to induce high mortality rates on larvae. Diversity and abundance of Vibrio varied significantly over time with highest values during and after spawning season. Vibrio communities from transplanted and stationary oysters converged over time, indicating that communities were not population specific, but rather assemble from the surrounding environment forming communities, some of which can persist over longer periods.

Wendling, Carolin C.; Batista, Frederico M.; Wegner, K. Mathias

2014-01-01

128

Challenge of New Zealand Greenshell mussel Perna canaliculus larvae using two Vibrio pathogens: a hatchery study.  

PubMed

Bacterial diseases remain a large problem in aquaculture hatcheries. The successful design and implementation of protective measures in the hatchery depends on an understanding of the dynamics of the infection process. Developing an in situ experimental protocol for pathogen challenge is therefore of paramount importance. Here, we demonstrated the minimum effective pathogenic dose (MEPD) of Vibrio splendidus (10(5) CFU ml(-1)) and a Vibrio coralliilyticus/neptunius-like isolate, Vibrio sp. DO1 (10(6) CFU ml(-1)), for New Zealand Greenshell mussel (GSM, Perna canaliculus) larvae during hatchery production. In a flow-through water hatchery system, larvae given 1 to 2 h of static water exposure to these pathogen doses showed respective average cumulative mortalities of 58 and 69% on the fourth day following pathogen exposure. After the 1 to 2 h static exposure, larvae were returned to flow-through water. Larvae exposed to a dosage one order of magnitude greater than the MEPD had higher mortalities of 73 and 96% for V. splendidus and Vibrio sp. DO1 respectively. These 4 levels of mortality were significantly greater than those of the non-exposed control larvae which respectively averaged 23 and 35% in experiments involving V. splendidus and Vibrio sp. DO1. Experiments were repeated 4 times to confirm reproducibility. After pathogen exposure, pathogens were detected in the larvae and tank water of treatments with dosages of > or =10(5) CFU ml(-1) (V. splendidus) and 10(6) CFU ml(-1) (Vibrio sp. DO1), but not in treatments with lower pathogen dosages. The challenge protocols are reproducible and provide an opportunity to assess measures for the protection of GSM larvae against infection in the hatchery environment. PMID:19899345

Kesarcodi-Watson, Aditya; Kaspar, Heinrich; Lategan, M Josie; Gibson, Lewis F

2009-09-01

129

Relationships between environmental factors and pathogenic Vibrios in the Northern Gulf of Mexico.  

PubMed

Although autochthonous vibrio densities are known to be influenced by water temperature and salinity, little is understood about other environmental factors associated with their abundance and distribution. Densities of culturable Vibrio vulnificus containing vvh (V. vulnificus hemolysin gene) and V. parahaemolyticus containing tlh (thermolabile hemolysin gene, ubiquitous in V. parahaemolyticus), tdh (thermostable direct hemolysin gene, V. parahaemolyticus pathogenicity factor), and trh (tdh-related hemolysin gene, V. parahaemolyticus pathogenicity factor) were measured in coastal waters of Mississippi and Alabama. Over a 19-month sampling period, vibrio densities in water, oysters, and sediment varied significantly with sea surface temperature (SST). On average, tdh-to-tlh ratios were significantly higher than trh-to-tlh ratios in water and oysters but not in sediment. Although tlh densities were lower than vvh densities in water and in oysters, the opposite was true in sediment. Regression analysis indicated that SST had a significant association with vvh and tlh densities in water and oysters, while salinity was significantly related to vibrio densities in the water column. Chlorophyll a levels in the water were correlated significantly with vvh in sediment and oysters and with pathogenic V. parahaemolyticus (tdh and trh) in the water column. Furthermore, turbidity was a significant predictor of V. parahaemolyticus density in all sample types (water, oyster, and sediment), and its role in predicting the risk of V. parahaemolyticus illness may be more important than previously realized. This study identified (i) culturable vibrios in winter sediment samples, (ii) niche-based differences in the abundance of vibrios, and (iii) predictive signatures resulting from correlations between environmental parameters and vibrio densities. PMID:20817802

Johnson, C N; Flowers, A R; Noriea, N F; Zimmerman, A M; Bowers, J C; DePaola, A; Grimes, D J

2010-11-01

130

Vibrios Associated with Litopenaeus vannamei Larvae, Postlarvae, Broodstock, and Hatchery Probionts  

PubMed Central

Several bacteriological surveys were performed from 1994 to 1996 at different Litopenaeus vannamei hatcheries (in Ecuador) and shrimp farms (in Mexico). Samples were taken from routine productions of healthy and diseased L. vannamei larvae, postlarvae, and their culture environment and from healthy and diseased juveniles and broodstock. In Ecuador, the dominant bacterial flora associated with shrimp larvae showing symptoms of zoea 2 syndrome, mysis mold syndrome, and bolitas syndrome has been determined. Strains were characterized by Biolog metabolic fingerprinting and identified by comparison to a database of 850 Vibrio type and reference strains. A selection of strains was further genotypically fine typed by AFLP. Vibrio alginolyticus is predominantly present in all larval stages and is associated with healthy nauplius and zoea stages. AFLP genetic fingerprinting shows high genetic heterogeneity among V. alginolyticus strains, and the results suggest that putative probiotic and pathogenic strains each have specific genotypes. V. alginolyticus was found to be associated with larvae with the zoea 2 syndrome and the mysis mold syndrome, while different Vibrio species (V. alginolyticus and V. harveyi) are associated with the bolitas syndrome. V. harveyi is associated with diseased postlarvae, juveniles, and broodstock. The identities of the strains identified as V. harveyi by the Biolog system could not be unambiguously confirmed by AFLP genomic fingerprinting. Vibrio strain STD3-988 and one unidentified strain (STD3-959) are suspected pathogens of only juvenile and adult stages. V. parahaemolyticus, Photobacterium damselae, and V. mimicus are associated with juvenile and adult stages.

Vandenberghe, Johan; Verdonck, Linda; Robles-Arozarena, Rocio; Rivera, Gabriel; Bolland, Annick; Balladares, Marcos; Gomez-Gil, Bruno; Calderon, Jorge; Sorgeloos, Patrick; Swings, Jean

1999-01-01

131

Vibrios associated with Litopenaeus vannamei larvae, postlarvae, broodstock, and hatchery probionts.  

PubMed

Several bacteriological surveys were performed from 1994 to 1996 at different Litopenaeus vannamei hatcheries (in Ecuador) and shrimp farms (in Mexico). Samples were taken from routine productions of healthy and diseased L. vannamei larvae, postlarvae, and their culture environment and from healthy and diseased juveniles and broodstock. In Ecuador, the dominant bacterial flora associated with shrimp larvae showing symptoms of zoea 2 syndrome, mysis mold syndrome, and bolitas syndrome has been determined. Strains were characterized by Biolog metabolic fingerprinting and identified by comparison to a database of 850 Vibrio type and reference strains. A selection of strains was further genotypically fine typed by AFLP. Vibrio alginolyticus is predominantly present in all larval stages and is associated with healthy nauplius and zoea stages. AFLP genetic fingerprinting shows high genetic heterogeneity among V. alginolyticus strains, and the results suggest that putative probiotic and pathogenic strains each have specific genotypes. V. alginolyticus was found to be associated with larvae with the zoea 2 syndrome and the mysis mold syndrome, while different Vibrio species (V. alginolyticus and V. harveyi) are associated with the bolitas syndrome. V. harveyi is associated with diseased postlarvae, juveniles, and broodstock. The identities of the strains identified as V. harveyi by the Biolog system could not be unambiguously confirmed by AFLP genomic fingerprinting. Vibrio strain STD3-988 and one unidentified strain (STD3-959) are suspected pathogens of only juvenile and adult stages. V. parahaemolyticus, Photobacterium damselae, and V. mimicus are associated with juvenile and adult stages. PMID:10347048

Vandenberghe, J; Verdonck, L; Robles-Arozarena, R; Rivera, G; Bolland, A; Balladares, M; Gomez-Gil, B; Calderon, J; Sorgeloos, P; Swings, J

1999-06-01

132

Power plays: iron transport and energy transduction in pathogenic vibrios  

PubMed Central

The Vibrios are a unique group of bacteria inhabiting a vast array of aquatic environments. Many Vibrio species are capable of infecting a wide assortment of hosts. Some of these species include V. parahaemolyticus, V. alginolyticus, V. vulnificus, V. anguillarum, and V. cholerae. The ability of these organisms to utilize iron is essential in establishing both an infection in their hosts as well as surviving in the environment. Bacteria are able to sequester iron through the secretion of low molecular weight iron chelators termed siderophores. The iron-siderophore complexes are bound by specific outer membrane receptors and are brought through both the outer and inner membranes of the cell. The energy needed to drive this active transport is achieved through the TonB energy transduction system. When first elucidated in E. coli, the TonB system was shown to be a three protein complex consisting of TonB, ExbB and ExbD. Most Vibrio species carry two TonB systems. The second TonB system includes a fourth protein; TtpC, which is essential for TonB2 mediated iron transport. Some Vibrio species have been shown to carry a third TonB system that also includes a TtpC protein.

Kustusch, Ryan J.; Kuehl, Carole J.; Crosa, Jorge H.

2011-01-01

133

Detection of pathogenic Vibrio parahaemolyticus in oyster enrichments by real time PCR  

Microsoft Academic Search

A real time polymerase chain reaction (PCR) assay was developed and evaluated to detect the presence of the thermostable direct hemolysin gene (tdh), a current marker of pathogenicity in Vibrio parahaemolyticus. The real time PCR fluorogenic probe and primer set was tested against a panel of numerous strains from 13 different bacterial species. Only V. parahaemolyticus strains possessing the tdh

George M Blackstone; Jessica L Nordstrom; Michael C. L Vickery; Michael D Bowen; Richard F Meyer; Angelo DePaola

2003-01-01

134

The Virulence Regulatory Protein ToxR Mediates Enhanced Bile Resistance in Vibrio cholerae and Other Pathogenic Vibrio Species  

PubMed Central

The transmembrane regulatory protein ToxR is required for expression of virulence factors in the human diarrheal pathogen Vibrio cholerae, including cholera toxin (CT) and the toxin coregulated pilus (TCP). ToxR is necessary for transcription of the gene encoding a second regulatory protein, ToxT, which is the direct transcriptional activator of CT and TCP genes. However, ToxR, independent of ToxT, directly activates and represses transcription of the outer membrane porins OmpU and OmpT, respectively. The genes encoding TCP and CT (and including ToxT) lie on horizontally acquired genetic elements, while the toxR, ompU, and ompT genes are apparently in the ancestral Vibrio chromosome. The contribution of ToxR-dependent modulation of outer membrane porins to cholera pathogenesis has remained unknown. We demonstrate that ToxR mediates enhanced bile resistance in a ToxT-independent manner. In both classical and El Tor biotypes of V. cholerae, a toxR mutant strain has a reduced minimum bactericidal concentration (MBC) of bile, the bile component deoxycholate (DC), and the anionic detergent sodium dodecyl sulfate (SDS) compared to both wild-type and toxT mutant strains. Classical and El Tor toxR mutant strains also exhibit reduced growth rates at subinhibitory concentrations of DC and SDS. Growth of either V. cholerae biotype in subinhibitory concentrations of bile or DC induces increased ToxR-dependent production of a major 38-kDa outer membrane protein, which was confirmed to be OmpU by Western blot. Measurement of transcription of a ompUp-lacZ fusion in both biotypes reveals stimulation (about two- to threefold) of ToxR-dependent ompU transcription by the presence of bile or DC, suggesting that ToxR may respond to the presence of bile. The toxR mutant strains of three additional human intestinal pathogenic Vibrio species, V. mimicus, V. fluvialis, and V. parahaemolyticus, display lower MBCs of bile, DC, and SDS and have altered outer membrane protein profiles compared to the parental wild-type strains. Our results demonstrate a conserved role for ToxR in the modulation of outer membrane proteins and bile resistance of pathogenic Vibrio species and suggest that these ToxR-dependent outer membrane proteins may mediate enhanced resistance to bile. We speculate that ToxR-mediated bile resistance was an early step in the evolution of V. cholerae as an intestinal pathogen.

Provenzano, Daniele; Schuhmacher, Darren A.; Barker, Justin L.; Klose, Karl E.

2000-01-01

135

Evolutionary Relationships of Pathogenic Clones of Vibrio cholerae by Sequence Analysis of Four Housekeeping Genes  

Microsoft Academic Search

Studies of the Vibrio cholerae population, using molecular typing techniques, have shown the existence of several pathogenic clones, mainly sixth-pandemic, seventh-pandemic, and U.S. Gulf Coast clones. However, the relationship of the pathogenic clones to environmental V. cholerae isolates remains unclear. A previous study to determine the phylogeny of V. cholerae by sequencing the asd (aspartate semialdehyde dehydrogenase) gene of V.

ROY BYUN; LIAM D. H. ELBOURNE; RUITING LAN; PETER R. REEVES

136

Ocean warming and spread of pathogenic vibrios in the aquatic environment.  

PubMed

Vibrios are among the most common bacteria that inhabit surface waters throughout the world and are responsible for a number of severe infections both in humans and animals. Several reports recently showed that human Vibrio illnesses are increasing worldwide including fatal acute diarrheal diseases, such as cholera, gastroenteritis, wound infections, and septicemia. Many scientists believe this increase may be associated with global warming and rise in sea surface temperature (SST), although not enough evidence is available to support a causal link between emergence of Vibrio infections and climate warming. The effect of increased SST in promoting spread of vibrios in coastal and brackish waters is considered a causal factor explaining this trend. Field and laboratory studies carried out over the past 40 years supported this hypothesis, clearly showing temperature promotes Vibrio growth and persistence in the aquatic environment. Most recently, a long-term retrospective microbiological study carried out in the coastal waters of the southern North Sea provided the first experimental evidence for a positive and significant relationship between SST and Vibrio occurrence over a multidecadal time scale. As a future challenge, macroecological studies of the effects of ocean warming on Vibrio persistence and spread in the aquatic environment over large spatial and temporal scales would conclusively support evidence acquired to date combined with studies of the impact of global warming on epidemiologically relevant variables, such as host susceptibility and exposure. Assessing a causal link between ongoing climate change and enhanced growth and spread of vibrios and related illness is expected to improve forecast and mitigate future outbreaks associated with these pathogens. PMID:23280498

Vezzulli, Luigi; Colwell, Rita R; Pruzzo, Carla

2013-05-01

137

Vibrio Fluvialis: An Unusual Enteric Pathogen of Increasing Public Health Concern  

PubMed Central

In developing countries, the fraction of treated wastewater effluents being discharged into watersheds have increased over the period of time, which have led to the deteriorations of the qualities of major rivers in developing nations. Consequently, high densities of disease causing bacteria in the watersheds are regularly reported including incidences of emerging Vibrio fluvialis. Vibrio fluvialis infection remains among those infectious diseases posing a potentially serious threat to public health. This paper addresses the epidemiology of this pathogen; pathogenesis of its disease; and its clinical manifestations in humans.

Igbinosa, Etinosa O.; Okoh, Anthony I.

2010-01-01

138

Contamination of community water sources by potentially pathogenic vibrios following sea water inundation.  

PubMed

Potentially pathogenic members of the Vibrionaceae family including Vibrio cholerae and Vibrio parahemolyticus were isolated from domestic sources of drinking water in coastal villages following sea water inundation during the tsunami in Southern India. Phenotypic and genotypic studies were done to confirm the identity and detection of toxins. Vibrio-gyr (gyrase B gene) was detected in all sixteen vibrio isolates. Toxin regulating genes i.e.: ctx gene, tdh gene, and trh gene, however were not detected in any of the strains, thereby ruling out presence of toxins which could endanger human life. Other potentially pathogenic bacteria Aeromonas and Plesiomonas were also isolated from hand pumps and wells, in a few localities. There was no immediate danger in the form of an outbreak or sporadic gastroenteritis at the time of the study. Timely chlorination and restoration of potable water supply to the flood affected population by governmental and nongovernmental agencies averted waterborne gastroenteritis. Assessment of quality of water and detection of potential virulent organisms is an important public health activity following natural disasters. This work highlights the importance of screening water sources for potentially pathogenic microorganisms after natural disasters to avert outbreaks of gastroenteritis and other infectious diseases. PMID:18697592

Kanungo, Reba; Shashikala; Karunasagar, I; Srinivasan, S; Sheela, Devi; Venkatesh, K; Anitha, P

2007-12-01

139

Fast detection of Vibrio species potentially pathogenic for mollusc.  

PubMed

Vibrio tasmaniensis, Vibrio splendidus and Vibrio neptunius species were distributed worldwide and associated with aquaculture and have been reported as the cause of diseases in aquatic organisms. Polyphasic analyses for bacterial identification are not feasible for routine diagnostic because of the time involved. The aim of this study is to design three PCR primer sets that can assist with fast detection of these species. They were designed from the 16S ribosomal RNA gene, and PCR conditions were found. Each PCR test successfully identified all the tested strains of each target species. The combined specificity of V. tasmaniensis and V. splendidus primer sets offered the best coverage (86%) in terms of separating target organisms from other related species. The primer set of V. tasmaniensis showed a lower sensitivity limit (500 fg of DNA) than the V. splendidus set (1 pg) and both sets gave positive amplification using homogenized tissues from inoculated clams, with 10(2) and 10(4) cfu/g of clam, respectively. The primer set of V. neptunius was highly specific, showing only cross-reaction with V. parahaemolyticus species from 44 tested species. Its sensitivity limit was 100 pg of DNA. A small number of biochemical tests were proposed concurrently with the PCR to differentiate the cross-reacting bacteria. The time of detection of the three tested species was reduced and the further affected animals can be diagnosed in a rapid fraction of time. The detection of virulent strains of V. tasmaniensis pointed to the risk of mollusc culture outbreaks. PMID:19640660

Lago, Estela Pérez; Nieto, Teresa Pérez; Farto Seguín, Rosa

2009-11-18

140

A survey for phosphoglucose isomerase with lysyl aminopeptidase activity in Vibrionaceae and non-Vibrio pathogens.  

PubMed

Phosphoglucose isomerase (PGI) with a novel lysyl aminopeptidase (LysAP) activity was recently purified and characterized from Vibrio vulnificus. We showed that it cleaves the amino-terminal lysyl residue from des-Arg(10)-kallidin to produce des-Arg(9)-bradykinin, suggesting that it plays a role in virulence. A survey was conducted to determine the presence of this potential virulence-enhancing enzyme among twenty-three halotolerant human and fish pathogens from eleven species within the Vibrionaceae family, including V. vulnificus, V. parahaemolyticus, V. cholerae, Aeromonas hydrophila, and Plesiomonas shigelloides. In addition, fourteen species of non-Vibrionaceae pathogens were screened for LysAP activity. Cell lysates were partially purified by anion exchange chromatography and fractions were screened for LysAP and isomerase activities. PGI-LysAP activity was detected in chromatographic fractions from all the Vibrio species tested, but was not detected in any of the non-Vibrionaceae pathogens. Levels of isomerase and LysAP activity correlated (R(2)=0.92) for nine strains of V. vulnificus. Since the Vibrionaceae represent an important family of human and fish pathogens, our identification of PGI-LysAP activity in a broad array of vibrios may lead to the development of improved analytical methods for their identification as well as interventions to reduce the high morbidity and mortality associated with some Vibrionaceae infections in clinical, veterinary, and aquaculture settings. PMID:15752701

Richards, Gary P; Parveen, Salina

2005-04-15

141

Administration of Bacillus subtilis strains in the rearing water enhances the water quality, growth performance, immune response, and resistance against Vibrio harveyi infection in juvenile white shrimp, Litopenaeus vannamei.  

PubMed

In this study, vegetative cell suspensions of two Bacillus subtilis strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU ml(-1) in the rearing water of shrimp (Litopenaeus vannamei) for eight weeks. Both probiotic groups showed a significant reduction of ammonia, nitrite and nitrate ions under in vitro and in vivo conditions. In comparison to untreated control group, final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR) and digestive enzymatic activity were significantly greater in the BM5 and BM8 groups. Significant differences for survival were recorded in the BM8 group as compared to the control. Eight weeks after the start of experiment, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 80%, whereas cumulative mortality of the shrimp that had been given probiotics was 36.7% with MB8 and 50% with MB5. Subsequently, real-time RT-PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and ?-1,3-glucan- binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was only significantly up-regulated in the BM5 group compared to the BM8 and control groups. These results suggest that administration of B. subtilis strains in the rearing water confers beneficial effects for shrimp aquaculture, considering water quality, growth performance, digestive enzymatic activity, immune response and disease resistance. PMID:24161773

Zokaeifar, Hadi; Babaei, Nahid; Saad, Che Roos; Kamarudin, Mohd Salleh; Sijam, Kamaruzaman; Balcazar, Jose Luis

2014-01-01

142

Description of a bacteriocinogenic plasmid in Beneckea harveyi.  

PubMed Central

A total of 795 strains of marine Vibrio species and Beneckea harveyi, a luminescent marine bacterium, were isolated from various sources in the area of Galveston Island, Tex., and screened for the production of bacteriocin-like substances. More than 8% of the Vibrio isolates produced low-molecular-weight (dialyzable) substances, which were lethal to a test strain of V. parahaemolyticus. Approximately 5% of the B. harveyi isolates produced higher-molecular-weight (nondialyzable) substances which were lethal to a test strain of B. harveyi. One of the B. harveyi strains (strain SY) produced a nondialyzable substance which was lethal to two of 39 strains of B. harveyi. The substance showed no activity toward 17 test strains drawn from the Vibrionaceae and Enterobacteriaceae. Strain SY showed no sensitivity to its own lethal agent and was shown by agarose gel electrophoresis and electron microscopy to harbor a single plasmid of 38 x 10(6) daltons. Variants of strain SY lacking the plasmid were produced by growth in the presence of the antibiotic novobiocin. These variants lacked both the ability to produce the lethal substance and the ability to survive in its presence. The lethal agent produced by strain SY is the first bacteriocin reported in marine bacteria. The term "harveyicin" is proposed to name this lethal substance. Images

McCall, J O; Sizemore, R K

1979-01-01

143

Outcomes of Infections of Sea Anemone Aiptasia pallida with Vibrio spp. Pathogenic to Corals.  

PubMed

Incidents of coral disease are on the rise. However, in the absence of a surrogate animal host, understanding of the interactions between coral pathogens and their hosts remains relatively limited, compared to other pathosystems of similar global importance. A tropical sea anemone, Aiptasia pallida, has been investigated as a surrogate model to study certain aspects of coral biology. Therefore, to test whether the utility of this surrogate model can be extended to study coral diseases, in the present study, we tested its susceptibility to common coral pathogens (Vibrio coralliilyticus and Vibrio shiloi) as well as polymicrobial consortia recovered from the Caribbean Yellow Band Disease (CYBD) lesions. A. pallida was susceptible to each of the tested pathogens. A. pallida responded to the pathogens with darkening of the tissues (associated with an increased melanization) and retraction of tentacles, followed by complete disintegration of polyp tissues. Loss of zooxanthellae was not observed; however, the disease progression pattern is consistent with the behavior of necrotizing pathogens. Virulence of some coral pathogens in Aiptasia was paralleled with their glycosidase activities. PMID:24619233

Zaragoza, William J; Krediet, Cory J; Meyer, Julie L; Canas, Gabriela; Ritchie, Kim B; Teplitski, Max

2014-08-01

144

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

PubMed Central

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.

Oliver, J D; Warner, R A; Cleland, D R

1982-01-01

145

Role of heme compounds and haptoglobin in Vibrio vulnificus pathogenicity.  

PubMed Central

An induced peritonitis model was employed in mice to determine whether heme-containing molecules enhance the lethality of infections by Vibrio vulnificus. The lethality of intraperitoneal (ip) inocula of the bacteria was increased by concurrent injections (ip) of hemoglobin, methemoglobin, or hematin, but not by myoglobin. Similar results were obtained in mice with phenylhydrazine-induced hemoglobinemia, in which after ip injections of V. vulnificus, a direct correlation between lethality and levels of plasma hemoglobin was observed. In vitro studies indicated that the growth of V. vulnificus, which was limited in an iron-poor medium, was enhanced by the addition of hemoglobin in a manner similar to an inorganic iron source, ferric ammonium citrate. These results suggest that V. vulnificus is capable of extracting iron from hemoglobin for use as a nutrilite, thereby promoting growth and increased lethality in the in vivo models. Further studies with human serum cultures demonstrated that the growth of V. vulnificus was not decreased when hemoglobin added to the serum was completely complexed with haptoglobin; these results are in opposition to those with cultures of Escherichia coli. These results are discussed relative to the capacity of V. vulnificus to produce fatal human infections.

Helms, S D; Oliver, J D; Travis, J C

1984-01-01

146

Role of heme compounds and haptoglobin in Vibrio vulnificus pathogenicity.  

PubMed

An induced peritonitis model was employed in mice to determine whether heme-containing molecules enhance the lethality of infections by Vibrio vulnificus. The lethality of intraperitoneal (ip) inocula of the bacteria was increased by concurrent injections (ip) of hemoglobin, methemoglobin, or hematin, but not by myoglobin. Similar results were obtained in mice with phenylhydrazine-induced hemoglobinemia, in which after ip injections of V. vulnificus, a direct correlation between lethality and levels of plasma hemoglobin was observed. In vitro studies indicated that the growth of V. vulnificus, which was limited in an iron-poor medium, was enhanced by the addition of hemoglobin in a manner similar to an inorganic iron source, ferric ammonium citrate. These results suggest that V. vulnificus is capable of extracting iron from hemoglobin for use as a nutrilite, thereby promoting growth and increased lethality in the in vivo models. Further studies with human serum cultures demonstrated that the growth of V. vulnificus was not decreased when hemoglobin added to the serum was completely complexed with haptoglobin; these results are in opposition to those with cultures of Escherichia coli. These results are discussed relative to the capacity of V. vulnificus to produce fatal human infections. PMID:6746093

Helms, S D; Oliver, J D; Travis, J C

1984-08-01

147

Taxonomic revision of Harveyi clade bacteria (family Vibrionaceae) based on analysis of whole genome sequences.  

PubMed

Use of inadequate methods for classification of bacteria in the so-called Harveyi clade (family Vibrionaceae, Gammaproteobacteria) has led to incorrect assignment of strains and proliferation of synonymous species. In order to resolve taxonomic ambiguities within the Harveyi clade and to test usefulness of whole genome sequence data for classification of Vibrionaceae, draft genome sequences of 12 strains were determined and analysed. The sequencing included type strains of seven species: Vibrio sagamiensis NBRC 104589(T), Vibrio azureus NBRC 104587(T), Vibrio harveyi NBRC 15634(T), Vibrio rotiferianus LMG 21460(T), Vibrio campbellii NBRC 15631(T), Vibrio jasicida LMG 25398(T), and Vibrio owensii LMG 25443(T). Draft genome sequences of strain LMG 25430, previously designated the type strain of [Vibrio communis], and two strains (MWB 21 and 090810c) from the 'beijerinckii' lineage were also determined. Whole genomes of two additional strains (ATCC 25919 and 200612B) that previously could not be assigned to any Harveyi clade species were also sequenced. Analysis of the genome sequence data revealed a clear case of synonymy between V. owensii and [V. communis], confirming an earlier proposal to synonymize both species. Both strains from the 'beijerinckii' lineage were classified as V. jasicida, while the strains ATCC 25919 and 200612B were classified as V. owensii and V. campbellii, respectively. We also found that two strains, AND4 and Ex25, are closely related to Harveyi clade bacteria, but could not be assigned to any species of the family Vibrionaceae. The use of whole genome sequence data for the taxonomic classification of the Harveyi clade bacteria and other members of the family Vibrionaceae is also discussed. PMID:23710045

Urbanczyk, Henryk; Ogura, Yoshitoshi; Hayashi, Tetsuya

2013-07-01

148

Occurrence of potentially pathogenic Vibrio species in raw, processed, and ready-to-eat seafood and seafood products  

Microsoft Academic Search

This study investigated the occurrence (by means of the presence-absence test) and level (by means of a plate count technique) of selected potentially pathogenic Vibrio species in processed and ready-to-eat seafood, and some raw seafood normally used as raw materials or ingredients in these products, that were commercially available in Chiang Mai, Thailand. The level of Vibrio in raw seafood

Thararat Chitov; Sutipong Wongdao

149

Parallel Quorum Sensing Systems Converge to Regulate Virulence in Vibrio cholerae  

Microsoft Academic Search

The marine bacterium Vibrio harveyi possesses two quorum sensing systems (System 1 and System 2) that regulate bioluminescence. Although the Vibrio cholerae genome sequence reveals that a V. harveyi-like System 2 exists, it does not predict the existence of a V. harveyi-like System 1 or any obvious quorum sensing-controlled target genes. In this report we identify and characterize the genes

Melissa B. Miller; Karen Skorupski; Derrick H. Lenz; Ronald K. Taylor; Bonnie L. Bassler

2002-01-01

150

Genomic and proteomic analyses of the coral pathogen Vibrio coralliilyticus reveal a diverse virulence repertoire  

PubMed Central

Vibrio coralliilyticus has been implicated as an important pathogen of coral species worldwide. In this study, the nearly complete genome of Vibrio coralliilyticus strain P1 (LMG23696) was sequenced and proteases implicated in virulence of the strain were specifically investigated. The genome sequence of P1 (5?513?256?bp in size) consisted of 5222 coding sequences and 58 RNA genes (53 tRNAs and at least 5 rRNAs). Seventeen metalloprotease and effector (vgrG, hlyA and hcp) genes were identified in the genome and expressed proteases were also detected in the secretome of P1. As the VcpA zinc-metalloprotease has been considered an important virulence factor of V. coralliilyticus, a vcpA deletion mutant was constructed to evaluate the effect of this gene in animal pathogenesis. Both wild-type and mutant (?vcpA) strains exhibited similar virulence characteristics that resulted in high mortality in Artemia and Drosophila pathogenicity bioassays and strong photosystem II inactivation of the coral dinoflagellate endosymbiont (Symbiodinium). In contrast, the ?vcpA mutant demonstrated higher hemolytic activity and secreted 18 proteins not secreted by the wild type. These proteins included four types of metalloproteases, a chitinase, a hemolysin-related protein RbmC, the Hcp protein and 12 hypothetical proteins. Overall, the results of this study indicate that V. coralliilyticus strain P1 has a diverse virulence repertoire that possibly enables this bacterium to be an efficient animal pathogen.

de O Santos, Eidy; Alves, Nelson; Dias, Graciela M; Mazotto, Ana Maria; Vermelho, Alane; Vora, Gary J; Wilson, Bryan; Beltran, Victor H; Bourne, David G; Le Roux, Frederique; Thompson, Fabiano L

2011-01-01

151

An Improved Detection and Quantification Method for the Coral Pathogen Vibrio coralliilyticus  

PubMed Central

DNA- and RNA-based PCR and reverse-transcription real-time PCR assays were developed for diagnostic detection of the vcpA zinc-metalloprotease implicated in the virulence of the coral pathogen Vibrio coralliilyticus. Both PCR methods were highly specific for V. coralliilyticus and failed to amplify strains of closely-related Vibrio species. The assays correctly detected all globally occurring V. coralliilyticus isolates including a newly-described isolate [TAV24] infecting gorgonians in the Mediterranean Sea and highlighted those isolates that had been potentially misidentified, in particular V. tubiashii strains ATCC 19105 and RE22, historically described as important oyster pathogens. The real-time assay is sensitive, detecting 10 gene copies and the relationships between gene copy number and cycle threshold (CT) were highly linear (R2?99.7). The real-time assay was also not affected by interference from non-target DNA. These assays are useful for rapid detection of V. coralliilyticus and monitoring of virulence levels in environmental samples, allowing for implementation of timely management steps to limit and possibly prevent losses due to V. coralliilyticus infection, as well as furthering investigations of factors affecting pathogenesis of this important marine pathogen.

Wilson, Bryan; Muirhead, Andrew; Bazanella, Monika; Huete-Stauffer, Carla; Vezzulli, Luigi; Bourne, David G.

2013-01-01

152

In vitro evaluation of antibacterial activity of Asparagopsis taxiformis from the Straits of Messina against pathogens relevant in aquaculture.  

PubMed

Ethanol extracts of Asparagopsis taxiformis collected from the Straits of Messina (Italy) were screened for antibacterial activity against pathogenic shellfish and fish bacteria previously isolated from local marine and brackish environments. Genetic labelling by DNA barcoding allowed us to identify the algal population as a biogeographical strain conspecific to A. taxiformis. The extract obtained in May showed the broadest antibacterial activity against all tested pathogenic bacteria, especially against Vibrio alginolyticus, Vibrio vulnificus and Aeromonas salmonicida subsp. salmonicida. Moderate activity was observed against Photobacterium damselae subsp. damselae and Photobacterium damselae subsp. piscicida, Salmonella sp., Vibrio cholerae, Vibrio harveyi and Vibrio parahaemolyticus. The absence of cytotoxic effects of active algal extracts was verified using trypan blue exclusion test on cells of digestive glands of Mytilus galloprovincialis. The results indicated that ethanol extracts of A. taxiformis could represent a source of antibacterial substances with potential use in aquaculture. PMID:22078751

Genovese, G; Faggio, C; Gugliandolo, C; Torre, A; Spanò, A; Morabito, M; Maugeri, T L

2012-02-01

153

Taxonomy of bacterial fish pathogens  

PubMed Central

Bacterial taxonomy has progressed from reliance on highly artificial culture-dependent techniques involving the study of phenotype (including morphological, biochemical and physiological data) to the modern applications of molecular biology, most recently 16S rRNA gene sequencing, which gives an insight into evolutionary pathways (= phylogenetics). The latter is applicable to culture-independent approaches, and has led directly to the recognition of new uncultured bacterial groups, i.e. "Candidatus", which have been associated as the cause of some fish diseases, including rainbow trout summer enteritic syndrome. One immediate benefit is that 16S rRNA gene sequencing has led to increased confidence in the accuracy of names allocated to bacterial pathogens. This is in marked contrast to the previous dominance of phenotyping, and identifications, which have been subsequently challenged in the light of 16S rRNA gene sequencing. To date, there has been some fluidity over the names of bacterial fish pathogens, with some, for example Vibrio anguillarum, being divided into two separate entities (V. anguillarum and V. ordalii). Others have been combined, for example V. carchariae, V. harveyi and V. trachuri as V. harveyi. Confusion may result with some organisms recognized by more than one name; V. anguillarum was reclassified as Beneckea and Listonella, with Vibrio and Listonella persisting in the scientific literature. Notwithstanding, modern methods have permitted real progress in the understanding of the taxonomic relationships of many bacterial fish pathogens.

2011-01-01

154

DNA cloning, characterization, and inhibition studies of an ?-carbonic anhydrase from the pathogenic bacterium Vibrio cholerae.  

PubMed

We have cloned, purified, and characterized an ?-carbonic anhydrase (CA, EC 4.2.1.1) from the human pathogenic bacterium Vibrio cholerae, VchCA. The new enzyme has significant catalytic activity, and an inhibition study with sulfonamides and sulfamates led to the detection of a large number of low nanomolar inhibitors, among which are methazolamide, acetazolamide, ethoxzolamide, dorzolamide, brinzolamide, benzolamide, and indisulam (KI values in the range 0.69-8.1 nM). As bicarbonate is a virulence factor of this bacterium and since ethoxzolamide was shown to inhibit the in vivo virulence, we propose that VchCA may be a target for antibiotic development, exploiting a mechanism of action rarely considered until now. PMID:23181552

Del Prete, Sonia; Isik, Semra; Vullo, Daniela; De Luca, Viviana; Carginale, Vincenzo; Scozzafava, Andrea; Supuran, Claudiu T; Capasso, Clemente

2012-12-13

155

Genome anatomy of the gastrointestinal pathogen, Vibrio parahaemolyticus of crustacean origin  

PubMed Central

Vibrio parahaemolyticus, an important human pathogen, is associated with gastroenteritis and transmitted through partially cooked seafood. It has become a major concern in the production and trade of marine food products. The prevalence of potentially virulent and pathogenic V. parahaemolyticus in raw seafood is of public health significance. Here we describe the genome sequence of a V. parahaemolyticus isolate of crustacean origin which was cultured from prawns in 2008 in Selangor, Malaysia (isolate PCV08-7). The next generation sequencing and analysis revealed that the genome of isolate PCV08-7 has closest similarity to that of V. parahaemolyticus RIMD2210633. However, there are certain unique features of the PCV08-7 genome such as the absence of TDH-related hemolysin (TRH), and the presence of HU-alpha insertion. The genome of isolate PCV08-7 encodes a thermostable direct hemolysin (TDH), an important virulence factor that classifies PCV08-7 isolate to be a serovariant of O3:K6 strain. Apart from these, we observed that there is certain pattern of genetic rearrangements that makes V. parahaemolyticus PCV08-7 a non-pandemic clone. We present detailed genome statistics and important genetic features of this bacterium and discuss how its survival, adaptation and virulence in marine and terrestrial hosts can be understood through the genomic blueprint and that the availability of genome sequence entailing this important Malaysian isolate would likely enhance our understanding of the epidemiology, evolution and transmission of foodborne Vibrios in Malaysia and elsewhere.

2013-01-01

156

Ocean acidification and host-pathogen interactions: blue mussels, Mytilus edulis, encountering Vibrio tubiashii.  

PubMed

Ocean acidification (OA) can shift the ecological balance between interacting organisms. In this study, we have used a model system to illustrate the interaction between a calcifying host organism, the blue mussel Mytilus edulis and a common bivalve bacterial pathogen, Vibrio tubiashii, with organisms being exposed to a level of acidification projected to occur by the end of the 21st century. OA exposures of the mussels were carried out in relative long-term (4 months) and short-term (4 days) experiments. We found no effect of OA on the culturability of V.?tubiashii, in broth or in seawater. OA inhibited mussel shell growth and impaired crystalline shell structures but did not appear to affect mussel immune parameters (i.e haemocyte counts and phagocytotic capacity). Despite no evident impact on host immunity or growth and virulence of the pathogen, V.?tubiashii was clearly more successful in infecting mussels exposed to long-term OA compared to those maintained under ambient conditions. Moreover, OA exposed V.?tubiashii increased their viability when exposed to haemocytes of OA-treated mussel. Our findings suggest that even though host organisms may have the capacity to cope with periods of OA, these conditions may alter the outcome of host-pathogen interactions, favouring the success of the latter. PMID:24147969

Asplund, Maria E; Baden, Susanne P; Russ, Sarah; Ellis, Robert P; Gong, Ningping; Hernroth, Bodil E

2014-04-01

157

Molecular epidemiology and genetic variation of pathogenic Vibrio parahaemolyticus in Peru.  

PubMed

Vibrio parahaemolyticus is a foodborne pathogen that has become a public health concern at the global scale. The epidemiological significance of V. parahaemolyticus infections in Latin America received little attention until the winter of 1997 when cases related to the pandemic clone were detected in the region, changing the epidemic dynamics of this pathogen in Peru. With the aim to assess the impact of the arrival of the pandemic clone on local populations of pathogenic V. parahaemolyticus in Peru, we investigated the population genetics and genomic variation in a complete collection of non-pandemic strains recovered from clinical sources in Peru during the pre- and post-emergence periods of the pandemic clone. A total of 56 clinical strains isolated in Peru during the period 1994 to 2007, 13 strains from Chile and 20 strains from Asia were characterized by Multilocus Sequence Typing (MLST) and checked for the presence of Variable Genomic Regions (VGRs). The emergence of O3:K6 cases in Peru implied a drastic disruption of the seasonal dynamics of infections and a shift in the serotype dominance of pathogenic V. parahaemolyticus. After the arrival of the pandemic clone, a great diversity of serovars not previously reported was detected in the country, which supports the introduction of additional populations cohabitating with the pandemic group. Moreover, the presence of genomic regions characteristic of the pandemic clone in other non-pandemic strains may represent early evidence of genetic transfer from the introduced population to the local communities. Finally, the results of this study stress the importance of population admixture, horizontal genetic transfer and homologous recombination as major events shaping the structure and diversity of pathogenic V. parahaemolyticus. PMID:23696906

Gavilan, Ronnie G; Zamudio, Maria L; Martinez-Urtaza, Jaime

2013-01-01

158

Molecular Epidemiology and Genetic Variation of Pathogenic Vibrio parahaemolyticus in Peru  

PubMed Central

Vibrio parahaemolyticus is a foodborne pathogen that has become a public health concern at the global scale. The epidemiological significance of V. parahaemolyticus infections in Latin America received little attention until the winter of 1997 when cases related to the pandemic clone were detected in the region, changing the epidemic dynamics of this pathogen in Peru. With the aim to assess the impact of the arrival of the pandemic clone on local populations of pathogenic V. parahaemolyticus in Peru, we investigated the population genetics and genomic variation in a complete collection of non-pandemic strains recovered from clinical sources in Peru during the pre- and post-emergence periods of the pandemic clone. A total of 56 clinical strains isolated in Peru during the period 1994 to 2007, 13 strains from Chile and 20 strains from Asia were characterized by Multilocus Sequence Typing (MLST) and checked for the presence of Variable Genomic Regions (VGRs). The emergence of O3:K6 cases in Peru implied a drastic disruption of the seasonal dynamics of infections and a shift in the serotype dominance of pathogenic V. parahaemolyticus. After the arrival of the pandemic clone, a great diversity of serovars not previously reported was detected in the country, which supports the introduction of additional populations cohabitating with the pandemic group. Moreover, the presence of genomic regions characteristic of the pandemic clone in other non-pandemic strains may represent early evidence of genetic transfer from the introduced population to the local communities. Finally, the results of this study stress the importance of population admixture, horizontal genetic transfer and homologous recombination as major events shaping the structure and diversity of pathogenic V. parahaemolyticus.

Gavilan, Ronnie G.; Zamudio, Maria L.; Martinez-Urtaza, Jaime

2013-01-01

159

Refined identification of Vibrio bacterial flora from Acanthasther planci based on biochemical profiling and analysis of housekeeping genes.  

PubMed

We used a polyphasic approach for precise identification of bacterial flora (Vibrionaceae) isolated from crown-of-thorns starfish (COTS) from Lizard Island (Great Barrier Reef, Australia) and Guam (U.S.A., Western Pacific Ocean). Previous 16S rRNA gene phylogenetic analysis was useful to allocate and identify isolates within the Photobacterium, Splendidus and Harveyi clades but failed in the identification of Vibrio harveyi-like isolates. Species of the V harveyi group have almost indistinguishable phenotypes and genotypes, and thus, identification by standard biochemical tests and 16S rRNA gene analysis is commonly inaccurate. Biochemical profiling and sequence analysis of additional topA and mreB housekeeping genes were carried out for definitive identification of 19 bacterial isolates recovered from sick and wild COTS. For 8 isolates, biochemical profiles and topA and mreB gene sequence alignments with the closest relatives (GenBank) confirmed previous 16S rRNA-based identification: V. fortis and Photobacterium eurosenbergii species (from wild COTS), and V natriegens (from diseased COTS). Further phylogenetic analysis based on topA and mreB concatenated sequences served to identify the remaining 11 V harveyi-like isolates: V. owensii and V. rotiferianus (from wild COTS), and V. owensii, V. rotiferianus, and V. harveyi (from diseased COTS). This study further confirms the reliability of topA-mreB gene sequence analysis for identification of these close species, and it reveals a wider distribution range of the potentially pathogenic V. harveyi group. PMID:22013751

Rivera-Posada, J A; Pratchett, M; Cano-Gomez, A; Arango-Gomez, J D; Owens, L

2011-09-01

160

Kinetics of adhesion of selected fish-pathogenic Vibrio strains of skin mucus of gilt-head sea bream (Sparus aurata L.).  

PubMed Central

The kinetics of adhesion of Vibrio strains isolated from diseased fish to skin mucus of gilt-head sea bream was studied. A modified Langmuir adsorption isotherm was calculated, and the results obtained indicate that the strains tested (Vibrio alginolyticus DP1HE4 and Vibrio anguillarum-like DC12R8 and DC12R9) showed a saturation kinetics except for V. alginolyticus (CAN), which showed a proportional adsorption kinetics. The adhesive capability for skin mucus does not seem to be an essential virulence factor of pathogenic strains of Vibrio, since this specific interaction depended on several environmental factors, temperature and salinity being the most important. However, the absence of an inhibitory effect of mucus on the pathogenic microorganisms, and the capability of the Vibrio strains to utilize mucus as a carbon source, could favor their settlement on the skin with a potential for infection of cultured, stressed fish.

Bordas, M A; Balebona, M C; Zorrilla, I; Borrego, J J; Morinigo, M A

1996-01-01

161

Disentangling the population structure and evolution of the clam pathogen Vibrio tapetis.  

PubMed

Vibrio tapetis is a fastidious slow-growing microorganism that causes the Brown Ring Disease in clams. Recently, two subspecies for this bacterial pathogen have been proposed. We have developed a multilocus sequence typing scheme and performed evolutionary studies of V. tapetis population using the great majority of isolates of V. tapetis obtained worldwide until now (30 isolates). V. tapetis constitutes a high polymorphic population, showing low diversity indexes and some genetic discontinuity among the isolates. Mutation events are more frequent than recombination, although both are approximately equally important for genetic diversification. In fact, the divergence between subspecies occurred exclusively by mutation but the diversity observed among isolates of the same subspecies appeared to be generated mostly by recombination. Between the subspecies, genetic distance is very high and almost no recurrent gene flow exists. This pathogen displays a non-clonal population structure with an ancient spatial segregation population and some degree of geographical isolation, followed by a population expansion, at least for V. tapetis subsp. tapetis. A database from this study was created and hosted on publmlst.org ( http://pubmlst.org/vtapetis/ ). PMID:24276536

Balboa, Sabela; Bastardo, Asmine; Romalde, Jesús L

2014-01-01

162

Non-coding sRNAs regulate virulence in the bacterial pathogen Vibrio cholerae  

PubMed Central

Vibrio cholerae is the waterborne bacterium responsible for worldwide outbreaks of the acute, potentially fatal cholera diarrhea. The primary factors this human pathogen uses to cause the disease are controlled by a complex regulatory program linking extracellular signaling inputs to changes in expression of several critical virulence genes. Recently it has been uncovered that many non-coding regulatory sRNAs are important components of the V. cholerae virulence regulon. Most of these sRNAs appear to require the RNA-binding protein, Hfq, to interact with and alter the expression of target genes, while a few sRNAs appear to function by an Hfq-independent mechanism. Direct base-pairing between the sRNAs and putative target mRNAs has been shown in a few cases but the extent of each sRNAs regulon is not fully known. Genetic and biochemical methods, coupled with computational and genomics approaches, are being used to validate known sRNAs and also to identify many additional putative sRNAs that may play a role in the pathogenic lifestyle of V. cholerae.

Bardill, J. Patrick; Hammer, Brian

2012-01-01

163

Pathogenicity of Vibrio alginolyticus for Cultured Gilt-Head Sea Bream (Sparus aurata L.)  

PubMed Central

The in vivo and in vitro pathogenic activities of whole cells and extracellular products of Vibrio alginolyticus for cultured gilt-head sea bream were evaluated. The 50% lethal doses ranged from 5.4 × 104 to 1.0 × 106 CFU/g of body weight. The strains examined had the ability to adhere to skin, gill, and intestinal mucus of sea bream and to cultured cells of a chinook salmon embryo cell line. In addition, the in vitro ability of V. alginolyticus to adhere to mucus and skin cells of sea bream was demonstrated by scanning electron microscopy. The biological activities of extracellular products of V. alginolyticus were hydrolytic activities; the products were able to degrade sea bream mucus. V. alginolyticus was cytotoxic for fish cell lines and lethal for sea bream. Moreover, the extracellular products could degrade sea bream tissues. However, experiments performed with the bath immersion inoculation technique demonstrated that V. alginolyticus should be considered a pathogen for sea bream only when the mucus layer is removed and the skin is damaged.

Balebona, M. Carmen; Andreu, Manuel J.; Bordas, M. Angeles; Zorrilla, Irene; Morinigo, Miguel A.; Borrego, Juan J.

1998-01-01

164

In Silico Analyses of Primers Used to Detect the Pathogenicity Genes of Vibrio cholerae  

PubMed Central

In Vibrio cholerae, the etiological agent of cholera, most of the virulence genes are located in two pathogenicity islands, named TCP (Toxin-Co-regulated Pilus) and CTX (Cholera ToXins). For each V. cholerae pathogenicity gene, we retrieved every primer published since 1990 and every known allele in order to perform a complete in silico survey and assess the quality of the PCR primers used for amplification of these genes. Primers with a melting temperature in the range 55–60°C against any target sequence were considered valid. Our survey clearly revealed that two thirds of the published primers are not able to properly detect every genetic variant of the target genes. Moreover, the quality of primers did not improve with time. Their lifetime, i.e. the number of times they were cited in the literature, is also not a factor allowing the selection of valid primers. We were able to improve some primers or design new primers for the few cases where no valid primer was found. In conclusion, many published primers should be avoided or improved for use in molecular detection tests, in order to improve and perfect specificity and coverage. This study suggests that bioinformatic analyses are important to validate the choice of primers.

Gardes, Julien; Croce, Olivier; Christen, Richard

2012-01-01

165

Complete Sequence of Virulence Plasmid pJM1 from the Marine Fish Pathogen Vibrio anguillarum Strain 775  

Microsoft Academic Search

The virulence plasmid pJM1 enables the fish pathogen Vibrio anguillarum, a gram-negative polarly flagel- lated comma-shaped rod bacterium, to cause a highly fatal hemorrhagic septicemic disease in salmonids and other fishes, leading to epizootics throughout the world. The pJM1 plasmid 65,009-nucleotide sequence, with an overall GC content of 42.6%, revealed genes and open reading frames (ORFs) encoding iron transporters, nonribosomal

Manuela Di Lorenzo; Michiel Stork; Marcelo E. Tolmasky; Luis A. Actis; David Farrell; Timothy J. Welch; Lidia M. Crosa; Anne M. Wertheimer; Qian Chen; Patricia Salinas; Lillian Waldbeser; Jorge H. Crosa

2003-01-01

166

Pathogenic potential of vibriophages against an experimental infection with Vibrio cholerae O1 in the RITARD model  

Microsoft Academic Search

Cholera continues to be an important public health problem in developing countries, including India. This study concerns the feasibility of possible exploitation of bacteriophages as a biocontrol agent to eliminate the pathogen Vibrio cholerae from the gut using the removable intestinal tie–adult rabbit diarrhoea (RITARD) model. A control rabbit challenged with 109 colony-forming units (CFU)\\/mL of V. cholerae MAK 757

Tushar Suvra Bhowmick; Hemanta Koley; Mayukh Das; Dhira Rani Saha; B. L. Sarkar

2009-01-01

167

Pathogenicity of Vibrio tapetis, the etiological agent of brown ring disease in clams.  

PubMed

Brown ring disease (BRD) causes high mortalities in the introduced Manila clam Ruditapes philippinarum cultured in western Europe. The etiological agent of BRD, Vibrio tapetis, adheres to and disrupts the production of the periostracal lamina, causing the anomalous deposition of periostracum around the inner shell. Because the primary sign of BRD is found outside the soft tissues, the processes leading to death are not as obvious as those for internal pathogens. This study was designed to evaluate the pathogenicity of V. tapetis, in an attempt to help explain the mechanisms of mortality. We found high mortalities (up to 100%) for clams following the inoculation of V. tapetis into the extrapallial space (between mantle and inner shell) or the posterior adductor muscle of healthy R. philippinarum. Microscopy and immunological detection methods showed that the pathogen was rapidly eliminated from tissues and hemolymph of animals that survived the inoculation. In clams that died, the bacteria were found to have proliferated, resulting in severe tissue disruption. Bacteria were able to penetrate into tissues from the extrapallial space through the external epithelium of the mantle. In contrast, no mortalities were observed following injection of V. tapetis in the native European clam Ruditapes decussatus, which is resistant to BRD. This clam rapidly eliminated the bacterium from hemolymph and soft tissues. Clam mortality associated with BRD in the field is likely to result from the penetration of V. tapetis into the clam's extrapallial space through the disrupted periostracal lamina and from there into the soft tissues through the irritated mantle epithelium. Some bacteria also penetrate through the digestive epithelia. In either case, bacteria proliferate rapidly in the soft tissues, causing severe damage and subsequent death. PMID:12033709

Allam, Bassem; Paillard, Christine; Ford, Susan E

2002-04-01

168

Identification and characterisation of pathogenic Vibrio splendidus from Yesso scallop (Patinopecten yessoensis) cultured in a low temperature environment.  

PubMed

A fatal disease with apparent lesions on the adductor muscles of Yesso scallop Patinopecten yessoensis has occurred for three consecutive years from 2009 to 2011 in the northeastern China. This disease has been suspected to be closely associated with bacterial infections. In the present study, 96 bacteria were isolated from the lesions of moribund scallops. They were grouped into Vibrio (88 strains), Moritella (5 strains), Pseudomonas (2 strains) and Shewanella (1 strain) by 16S rDNA sequencing. In the Vibrio group, Vibrio splendidus was the predominant species, consisting of 43 strains. A strain of V. splendidus, named JZ6, was identified as a potential pathogen because it possessed two important virulence factors, Vsm and OmpU. Stain JZ6 was cold-adapted, and could surviveand exhibit haemolytic activity from 4°C to 32°C, with the highest activity observed at 10°C. The pathogenicity of JZ6 was further confirmed by injection and immersion challenges to scallops at 10°C. The cumulative mortality rates in the two experiments were 80% and 45%, respectively, which were significantly higher (P<0.05) than those of control and blank groups. The disorganisation of muscle fibres in challenged scallops was observed to be histopathologically identical to that in moribund Yesso scallop collected from the same farm. The results indicate that V. splendidus is the predominantly culturable bacteria from the lesions of diseased Yesso scallop, and JZ6 probably is the pathogenic agent to Yesso scallop in this low temperature environment. PMID:23911357

Liu, Rui; Qiu, Limei; Yu, Zuoan; Zi, Jun; Yue, Feng; Wang, Lingling; Zhang, Huan; Teng, Weiming; Liu, Xiangfeng; Song, Linsheng

2013-10-01

169

Interactions between the pathogenic bacterium Vibrio parahaemolyticus and red-tide dinoflagellates  

NASA Astrophysics Data System (ADS)

Vibrio parahaemolyticus is a common pathogenic bacterium in marine and estuarine waters. To investigate interactions between V. parahaemolyticus and co-occurring redtide dinoflagellates, we monitored the daily abundance of 5 common red tide dinoflagellates in laboratory culture; Amphidinium carterae, Cochlodinium ploykrikoides, Gymnodinium impudicum, Prorocentrum micans, and P. minimum. Additionally, we measured the ingestion rate of each dinoflagellate on V. parahaemolyticus as a function of prey concentration. Each of the dinoflagellates responded differently to the abundance of V. parahaemolyticus. The abundances of A. carterae and P. micans were not lowered by V. parahaemolyticus, whereas that of C. polykrikodes was lowered considerably. The harmful effect depended on bacterial concentration and incubation time. Most C. polykrikoides cells died after 1 hour incubation when the V. parahaemolyticus concentration was 1.4×107 cells ml-1, while cells died within 2 days of incubation when the bacterial concentration was 1.5×106 cells ml-1. With increasing V. parahaemolyticus concentration, ingestion rates of P. micans, P. minimum, and A. carterae on the prey increased, whereas that on C. polykrikoides decreased. The maximum or highest ingestion rates of P. micans, P. minimum, and A. carterae on V. parahaemolyticus were 55, 5, and 2 cells alga-1 h-1, respectively. The results of the present study suggest that V. parahaemolyticus can be both the killer and prey for some red tide dinoflagellates.

Seong, Kyeong Ah; Jeong, Hae Jin

2011-06-01

170

Two pathogens of Greenshell mussel larvae, Perna canaliculus: Vibrio splendidus and a V. coralliilyticus/neptunius-like isolate.  

PubMed

Bacterial pathogens of Greenshell mussel (GSM) larvae can cause batch losses during hatchery production. Twenty-two isolates were screened using a larval bioassay. Two strains were identified as potential pathogens. Phenotypic identification of these strains revealed two non-reactive Gram-negative, oxidase positive rods. Sequencing of the 16S rRNA gene identified Vibrio splendidus and a V. coralliilyticus/neptunius-like isolate as pathogens of GSM larvae, with an ability to cause 83% and 75% larval mortality in vitro, respectively, at a concentration of 10(2) CFU mL(-1). Histopathology indicated that the route of infection was via the digestive system. Using healthy larvae as target hosts, Koch's postulates were confirmed for the two isolates. This is the first report on pathogens of GSM larvae. PMID:19538643

Kesarcodi-Watson, A; Kaspar, H; Lategan, M J; Gibson, L

2009-06-01

171

Microarray-based detection of genetic heterogeneity, antimicrobial resistance, and the viable but nonculturable state in human pathogenic Vibrio spp.  

PubMed

The morbidity and mortality associated with Vibrio-mediated waterborne diseases necessitates the development of sensitive detection technologies that are able to elucidate the identity, potential pathogenicity, susceptibility, and viability of contaminating bacteria in a timely manner. For this purpose, we have designed a single multiplex PCR assay to simultaneously amplify 95 diagnostic regions (encompassing species/serogroup-specific, antimicrobial resistance, and known toxin markers) and combined it with a long oligonucleotide microarray to create a platform capable of rapidly detecting and discriminating the major human pathogenic species from the genus Vibrio: V. cholerae, V. parahaemolyticus, V. vulnificus, and V. mimicus. We were able to validate this strategy by testing 100 geographically and temporally distributed isolates and observed an excellent concordance between species- and serotype-level microarray-based identification and traditional typing methods. In addition to accurate identification, the microarray simultaneously provided evidence of antibiotic resistance genes and mobile genetic elements, such as sulfamethoxazole-trimethoprim constins and class I integrons, and common toxin (ctxAB, rtxA, hap, hlyA, tl, tdh, trh, vvhA, vlly, and vmhA) and pathogenicity (tcpA, type III secretion system) genes that are associated with pathogenic Vibrio. The versatility of this method was further underscored by its ability to detect the expression of known toxin and virulence genes from potentially harmful viable but nonculturable organisms. The results suggest that this molecular identification method provides rapid and definitive information that would be of value in epidemiological, environmental, and health risk assessment surveillance. PMID:16354840

Vora, Gary J; Meador, Carolyn E; Bird, Michele M; Bopp, Cheryl A; Andreadis, Joanne D; Stenger, David A

2005-12-27

172

Genes Similar to the Vibrio parahaemolyticus Virulence-Related Genes tdh, tlh, and vscC2 Occur in Other Vibrionaceae Species Isolated from a Pristine Estuary  

PubMed Central

Detection of the human pathogen Vibrio parahaemolyticus often relies on molecular biological analysis of species-specific virulence factor genes. These genes have been employed in determinations of V. parahaemolyticus population numbers and the prevalence of pathogenic V. parahaemolyticus strains. Strains of the Vibrionaceae species Photobacterium damselae, Vibrio diabolicus, Vibrio harveyi, and Vibrio natriegens, as well as strains similar to Vibrio tubiashii, were isolated from a pristine salt marsh estuary. These strains were examined for the V. parahaemolyticus hemolysin genes tdh, trh, and tlh and for the V. parahaemolyticus type III secretion system 2? gene vscC2 using established PCR primers and protocols. Virulence-related genes occurred at high frequencies in non-V. parahaemolyticus Vibrionaceae species. V. diabolicus was of particular interest, as several strains were recovered, and the large majority (>83%) contained virulence-related genes. It is clear that detection of these genes does not ensure correct identification of virulent V. parahaemolyticus. Further, the occurrence of V. parahaemolyticus-like virulence factors in other vibrios potentially complicates tracking of outbreaks of V. parahaemolyticus infections.

Klein, Savannah L.; Gutierrez West, Casandra K.; Mejia, Diana M.

2014-01-01

173

Isolation and Characterization of Pathogenic Vibrio harveyi ( V. carchariae ) From the Farmed Marine Cobia Fish Rachycentron canadum L. with Gastroenteritis Syndrome  

Microsoft Academic Search

An outbreak of serious mortality among the cultivated juvenile cobia Rachycentron canadum L. (weighing 8–10 g) characterized by a swollen intestine containing transparent yellow fluid (ascites and gastroenteritis) occurred in August 2001 in Taiwan. Ten motile bacterial strains, C3d1–C3d10, were isolated from head kidney (an organ located near the head of the fish) and\\/or the intestinal yellow fluid on tryptic

Ping-Chung Liu; Ji-Yang Lin; Wen-Hsiao Chuang; Kuo-Kau Lee

2004-01-01

174

Functional Characterization of Two Type III Secretion Systems of Vibrio parahaemolyticus  

PubMed Central

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.

Park, Kwon-Sam; Ono, Takahiro; Rokuda, Mitsuhiro; Jang, Myoung-Ho; Okada, Kazuhisa; Iida, Tetsuya; Honda, Takeshi

2004-01-01

175

Vibrio infections triggering mass mortality events in a warming Mediterranean Sea.  

PubMed

Mass mortality events of benthic invertebrates in the temperate north-western (NW) Mediterranean Sea have been observed in recent seasons. A 16 month in situ study in the Ligurian Sea (NW Mediterranean Sea) demonstrated that the occurrence of Paramuricea clavata mortality episodes were concomitant to a condition of prolonged high sea surface temperatures, low chlorophyll concentrations and the presence of culturable Vibrio spp. in seawater. The occurrence of Vibrio spp. at the seasonal scale was correlated with temperature; with few vibrios retrieved on specific media when the temperature dropped below 18 degrees C and a sharp increase of vibrios abundance (up to 3.4 x 10(4) MPN l(-1)) when the temperature was greater than or equal to 22 degrees C. Phylogenetic and phenotypic analysis of Vibrio isolates associated with healthy and diseased P. clavata colonies collected during a mortality episode showed that these bacteria were significantly more abundant in diseased than in healthy corals and were related to the V. harveyi, V. splendidus and V. coralliilyticus groups, the latter only identified in diseased organisms. Inoculation of bacterial isolates from these groups onto healthy P. clavata in aquaria caused disease signs and death in a range of Vibrio concentrations, temperature values and trophic conditions consistent with those recorded in the field. It is concluded that Vibrio infections may act as an additional triggering mechanism of mass mortality events in the coastal Mediterranean Sea and that their occurrence is climate-linked. Predicted global warming leading to long-lasting hot summer periods together with stratification resulting in energetic constraints represent a major threat to the survival of benthic invertebrates in the temperate NW Mediterranean Sea due to potential disease outbreak associated with Vibrio pathogens. PMID:20370818

Vezzulli, Luigi; Previati, Monica; Pruzzo, Carla; Marchese, Anna; Bourne, David G; Cerrano, Carlo

2010-07-01

176

Antibiofilm activity of Dendrophthoe falcata against different bacterial pathogens.  

PubMed

Dendrophthoe falcata is a hemiparasitic plant commonly used for ailments such as ulcers, asthma, impotence, paralysis, skin diseases, menstrual troubles, pulmonary tuberculosis, and wounds. In this context, the validations of the traditional claim that the leaf extract of D. falcata possesses antibiofilm and anti-quorum sensing activity against different bacterial pathogens were assessed. The bacterial biofilms were quantified by crystal violet staining. Among the 17 bacterial pathogens screened, the methanolic fraction of the leaf extract clearly demonstrated antibiofilm activity for Proteus mirabilis, Vibrio vulnificus, Aeromonas hydrophila, Shigella sonnei, Chromobacterium violaceum ATCC 12472, Vibrio parahaemolyticus, Vibrio harveyi, Vibrio alginolyticus, Vibrio cholerae, and Proteus vulgaris. At biofilm inhibitory concentrations, biofilm formation was reduced by up to 70-90?%. Furthermore, the potential quorum-sensing activity of the leaf extract was tested by agar well diffusion using Chromobacterium violaceum (ATCC 12472 & CV O26) reporter strains. The inhibition of violacein production may be due to direct or indirect interference on QS by active constituents or the interactive effect of different phytocompounds present in the extracts. This is the first report on antibiofilm and QS activity of D. falcata leaf extracts, signifying the scope for development of complementary medicine for biofilm-associated infections. PMID:23115018

Karthikeyan, Alagarsamy; Rameshkumar, Ramakrishnan; Sivakumar, Nallusamy; Al Amri, Issa S; Karutha Pandian, Shunmugiah; Ramesh, Manikandan

2012-12-01

177

Use of OmpU porins for attachment and invasion of Crassostrea gigas immune cells by the oyster pathogen Vibrio splendidus  

PubMed Central

OmpU porins are increasingly recognized as key determinants of pathogenic host Vibrio interactions. Although mechanisms remain incompletely understood, various species, including the human pathogen Vibrio cholera, require OmpU for host colonization and virulence. We have shown previously that OmpU is essential for virulence in the oyster pathogen Vibrio splendidus LGP32. Here, we showed that V. splendidus LGP32 invades the oyster immune cells, the hemocytes, through subversion of host-cell actin cytoskeleton. In this process, OmpU serves as an adhesin/invasin required for ?-integrin recognition and host cell invasion. Furthermore, the major protein of oyster plasma, the extracellular superoxide dismutase Cg-EcSOD, is used as an opsonin mediating the OmpU-promoted phagocytosis through its RGD sequence. Finally, the endocytosed bacteria were found to survive intracellularly, evading the host defense by preventing acidic vacuole formation and limiting reactive oxygen species production. We conclude that (i) V. splendidus is a facultative intracellular pathogen that manipulates host defense mechanisms to enter and survive in host immune cells, and (ii) that OmpU is a major determinant of host cell invasion in Vibrio species, used by V. splendidus LGP32 to attach and invade oyster hemocytes through opsonisation by the oyster plasma Cg-EcSOD.

Duperthuy, Marylise; Schmitt, Paulina; Garzon, Edwin; Caro, Audrey; Rosa, Rafael D.; Le Roux, Frederique; Lautredou-Audouy, Nicole; Got, Patrice; Romestand, Bernard; de Lorgeril, Julien; Kieffer-Jaquinod, Sylvie; Bachere, Evelyne; Destoumieux-Garzon, Delphine

2011-01-01

178

Use of OmpU porins for attachment and invasion of Crassostrea gigas immune cells by the oyster pathogen Vibrio splendidus.  

PubMed

OmpU porins are increasingly recognized as key determinants of pathogenic host Vibrio interactions. Although mechanisms remain incompletely understood, various species, including the human pathogen Vibrio cholera, require OmpU for host colonization and virulence. We have shown previously that OmpU is essential for virulence in the oyster pathogen Vibrio splendidus LGP32. Here, we showed that V. splendidus LGP32 invades the oyster immune cells, the hemocytes, through subversion of host-cell actin cytoskeleton. In this process, OmpU serves as an adhesin/invasin required for ?-integrin recognition and host cell invasion. Furthermore, the major protein of oyster plasma, the extracellular superoxide dismutase Cg-EcSOD, is used as an opsonin mediating the OmpU-promoted phagocytosis through its RGD sequence. Finally, the endocytosed bacteria were found to survive intracellularly, evading the host defense by preventing acidic vacuole formation and limiting reactive oxygen species production. We conclude that (i) V. splendidus is a facultative intracellular pathogen that manipulates host defense mechanisms to enter and survive in host immune cells, and (ii) that OmpU is a major determinant of host cell invasion in Vibrio species, used by V. splendidus LGP32 to attach and invade oyster hemocytes through opsonisation by the oyster plasma Cg-EcSOD. PMID:21282662

Duperthuy, Marylise; Schmitt, Paulina; Garzón, Edwin; Caro, Audrey; Rosa, Rafael D; Le Roux, Frédérique; Lautrédou-Audouy, Nicole; Got, Patrice; Romestand, Bernard; de Lorgeril, Julien; Kieffer-Jaquinod, Sylvie; Bachère, Evelyne; Destoumieux-Garzón, Delphine

2011-02-15

179

Examination of Diverse Toxin-Coregulated Pilus-Positive Vibrio cholerae Strains Fails To Demonstrate Evidence for Vibrio Pathogenicity Island Phage  

PubMed Central

The major virulence factors of toxigenic Vibrio cholerae are cholera toxin, which is encoded by a lysogenic filamentous bacteriophage (CTX?), and toxin-coregulated pilus (TCP), an essential colonization factor that is also the receptor for CTX?. The genes involved in the biosynthesis of TCP reside in a pathogenicity island, which has been reported to correspond to the genome of another filamentous phage (designated VPI?) and to encode functions necessary for the production of infectious VPI? particles. We examined 46 V. cholerae strains having diverse origins and carrying different genetic variants of the TCP island for the production of the VPI? and CTX? in different culture conditions, including induction of prophages with mitomycin C and UV irradiation. Although 9 of 10 V. cholerae O139 strains and 12 of 15 toxigenic El Tor strains tested produced extracellular CTX?, none of the 46 TCP-positive strains produced detectable VPI? in repeated assays, which detected as few as 10 particles of a control CTX phage per ml. These results contradict the previous report regarding VPI?-mediated horizontal transfer of the TCP genes and suggest that the TCP island is unable to support the production of phage particles. Further studies are necessary to understand the mechanism of horizontal transfer of the TCP island.

Faruque, Shah M.; Zhu, Jun; Asadulghani; Kamruzzaman, M.; Mekalanos, John J.

2003-01-01

180

Low incidence of Vibrio vulnificus among Vibrio isolates from sea water and shellfish of the western Mediterranean coast.  

PubMed

A specific search for Vibrio vulnificus in natural marine samples from the Spanish Mediterranean Sea was carried out by nested PCR and cultural approaches using thiosulphate-citrate-bile salts-sucrose agar (TCBS) and cellobiose-polymixin B-colistin agar (CPC), incubated at 40 degrees C, as selective media. Presumptive colonies were identified by PCR using specific primers against 23S rRNA sequences. This species was isolated from sea water and edible bivalves, mainly after preenrichment in alkaline peptone water (APW) at 40 degrees C followed by CPC agar. None of the V. vulnificus isolates identified corresponded to serovar E. Dominant Vibrio species on directly inoculated TCBS plates incubated at 25 degrees C were V. splendidus below 20 degrees C and V. harveyi and V. mediterranei above that temperature. Low percentages of several pathogenic vibrios were recorded but V. vulnificus was never recovered at this incubation temperature. The incidence of this species in the samples studied was lower than that described for other geographical areas, probably due to the high salinity values of the Mediterranean Sea. PMID:10030016

Arias, C R; Macián, M C; Aznar, R; Garay, E; Pujalte, M J

1999-01-01

181

Selected Bacterial Strains Protect Artemia spp. from the Pathogenic Effects of Vibrio proteolyticus CW8T2  

PubMed Central

In this study Vibrio proteolyticus CW8T2 has been identified as a virulent pathogen for Artemia spp. Its infection route has been visualized with transmission electron microscopy. The pathogen affected microvilli and gut epithelial cells, disrupted epithelial cell junctions, and reached the body cavity, where it devastated cells and tissues. In vivo antagonism tests showed that preemptive colonization of the culture water with nine selected bacterial strains protected Artemia juveniles against the pathogenic effects. Two categories of the selected strains could be distinguished: (i) strains providing total protection, as no mortality occurred 2 days after the experimental infection with V. proteolyticus CW8T2, with strain LVS8 as a representative, and (ii) strains providing partial protection, as significant but not total mortality was observed, with strain LVS2 as a representative. The growth of V. proteolyticus CW8T2 in the culture medium was slowed down in the presence of strains LVS2 and LVS8, but growth suppression was distinctly higher with LVS8 than with LVS2. It was striking that the strains that gave only partial protection against the pathogen in the in vivo antagonism test showed also a restricted capability to colonize the Artemia compared to the strains providing total protection. The in vivo antagonism tests and the filtrate experiments showed that probably no extracellular bacterial compounds were involved in the protective action but that the living cells were required to protect Artemia against V. proteolyticus CW8T2.

Verschuere, Laurent; Heang, Hanglamong; Criel, Godelieve; Sorgeloos, Patrick; Verstraete, Willy

2000-01-01

182

Selection and identification of non-pathogenic bacteria isolated from fermented pickles with antagonistic properties against two shrimp pathogens  

PubMed Central

In this study, potential probiotic strains were isolated from fermented pickles based on antagonistic activity against two shrimp pathogens (Vibrio harveyi and Vibrio parahaemolyticus). Two strains L10 and G1 were identified by biochemical tests, followed by16S ribosomal RNA gene sequence analysis as Bacillus subtilis, and characterized by PCR amplification of repetitive bacterial DNA elements (Rep-PCR). Subsequently, B. subtilis L10 and G1 strains were tested for antibacterial activity under different physical conditions, including culture medium, salinity, pH and temperature using the agar well diffusion assay. Among the different culture media, LB broth was the most suitable medium for antibacterial production. Both strains showed the highest level of antibacterial activity against two pathogens at 30?°C and 1.0% NaCl. Under the pH conditions, strain G1 showed the greatest activity against V. harveyi at pH 7.3–8.0 and against V. parahaemolyticus at pH 6.0–8.0, whereas strain L10 showed the greatest activity against two pathogens at pH 7.3. The cell-free supernatants of both strains were treated with four different enzymes in order to characterize the antibacterial substances against V. harveyi. The result showed considerable reduction of antibacterial activity for both strains, indicating the proteinaceous nature of the antibacterial substances. A wide range of tolerance to NaCl, pH and temperature was also recorded for both strains. In addition, both strains showed no virulence effect in juvenile shrimp Litopenaeus vannamei. On the basis of these results and safety of strains to L. vannamei, they may be considered for future challenge experiments in shrimp as a very promising alternative to the use of antibiotics.

Zokaeifar, Hadi; Luis Balcazar, Jose; Kamarudin, Mohd Salleh; Sijam, Kamaruzaman; Arshad, Aziz; Saad, Che Roos

2012-01-01

183

Evaluation of Global Gene Expression During Cold Shock in the Human Pathogen Vibrio vulnificus  

Microsoft Academic Search

Vibrio vulnificus can adapt to cold temperatures by changing the expression profiles of certain genes and their resulting proteins. In this\\u000a study, the complete V. vulnificus transcriptome was analyzed under cold shock by looking at gene expression changes occurring during the shift from 35°C to\\u000a 4°C. A DNA microarray-based global transcript profiling of V. vulnificus showed that 165 genes out

Raphael R. Wood; Cova R. Arias

184

Variability of Total and Pathogenic Vibrio parahaemolyticus Densities in Northern Gulf of Mexico Water and Oysters  

Microsoft Academic Search

Vibrio parahaemolyticus is indigenous to coastal environments and a frequent cause of seafood-borne gas- troenteritis in the United States, primarily due to raw-oyster consumption. Previous seasonal-cycle studies of V. parahaemolyticus have identified water temperature as the strongest environmental predictor. Salinity has also been identified, although it is evident that its effect on annual variation is not as pronounced. The effects

A. M. Zimmerman; A. DePaola; J. C. Bowers; J. A. Krantz; J. L. Nordstrom; C. N. Johnson; D. J. Grimes

2007-01-01

185

Electronic deoxyribonucleic acid (DNA) microarray detection of viable pathogenic Escherichia coli, Vibrio cholerae, and Salmonella typhi  

Microsoft Academic Search

An electronic deoxyribonucleic acid (DNA) microarray technique was developed for detection and identification of viable Escherichia coli O157:H7, Vibrio cholerae O1, and Salmonella typhi. Four unique genes, the E. coli O157 lipopolysaccharide (LPS) gene (rfbE) and H7 flagellin gene (fliC), the V. cholerae O1 LPS gene (rfbE), and the S. typhi LPS gene (tyv), were chosen as the targets for

Yanming Liu; Zhilong Gong; Nicole Morin; Odell Pui; Michael Cheung; Hai Zhang; Xing-Fang Li

2006-01-01

186

Pathogenic Potential of Non-O1, Non-O139 Vibrio cholerae  

Microsoft Academic Search

\\u000a \\u000a Vibrio cholerae organisms are known to exist in more than 200 different serogroups based on their “O” antigenic characters. Out of these,\\u000a strains belonging to O1 or O139 serogroups have been implicated as the causative agent of cholera in the epidemic form. On\\u000a the other hand, V. cholerae non-O1, non-O139 organisms, ubiquitously present in the aquatic environment, are usually nonpathogenic

Amit Sarkar; Ranjan K. Nandy; Asoke C. Ghose

187

Development of monoclonal antibodies that identify Vibrio species commonly isolated from infections of humans, fish, and shellfish.  

PubMed Central

Monoclonal antibodies (MAbs) against Vibrio species that infect humans, fish, and shellfish were developed for application in rapid identifications. The pathogens included Vibrio alginolyticus, V. anguillarum, V. carchariae, V. cholerae, V. damsela, V. furnissii, V. harveyi, V. ordalii, V. parahaemolyticus, and V. vulnificus. Three types of MAbs were selected. The first important group included MAbs that reacted with only a single species. A second group comprised a number of MAbs that reacted with two, taxonomically closely related Vibrio species. For example, of 22 MAbs raised against V. alginolyticus, 6 recognized a 52-kDa flagellar H antigen common to both V. alginolyticus and V. parahaemolyticus; V. anguillarum and V. ordalii also shared antigens. A third group included three genus-specific MAbs that reacted with almost all Vibrio species but did not react with other members of the family Vibrionaceae (e.g., members of the Aeromonas, Photobacterium, and Plesiomonas genera) or a wide range of gram-negative bacteria representing many genera. This last group indicated the possible existence of an antigenic determinant common to Vibrio species. Two of these three genus-specific MAbs reacted with heat-stable antigenic determinants of Vibrio species as well as lipopolysaccharide extracted from Vibrio species. The use of the MAbs in blind tests and diagnosis of clinical isolates indicated that three different types of bacteria, viz., live, formalin-fixed, and sodium azide-killed bacteria, were detected consistently. Overall, it was found that the genus-specific MAbs were very useful for rapidly identifying vibrios in the screening of acute infections, while the species-specific MAbs and others were useful for completing the diagnosis. Images

Chen, D; Hanna, P J; Altmann, K; Smith, A; Moon, P; Hammond, L S

1992-01-01

188

Epibiotic Vibrio luminous bacteria isolated from some hydrozoa and bryozoa species.  

PubMed

Luminous bacteria are isolated from both Hydrozoa and Bryozoa with chitinous structures on their surfaces. All the specimens of the examined hydroid species (Aglaophenia kirchenpaueri, Aglaophenia octodonta, Aglaophenia tubiformis, Halopteris diaphana, Plumularia setacea, Ventromma halecioides), observed under blue light excitation, showed a clear fluorescence on the external side of the perisarc (chitinous exoskeleton) around hydrocladia. In the bryozoan Myriapora truncata, luminous bacteria are present on the chitinous opercula. All the isolated luminous bacteria were identified on the basis of both phenotypic and genotypic analysis. The isolates from A. tubiformis and H. diaphana were unambiguously assigned to the species Vibrio fischeri. In contrast, the isolates from the other hydroids, phenotypically assigned to the species Vibrio harveyi, were then split into two distinct species by phylogenetic analysis of 16S rRNA gene sequences and DNA-DNA hybridization experiments. Scanning electron microscopy analysis and results of culture-based and culture-independent approaches enabled us to establish that luminous vibrios represent major constituents of the bacterial community inhabiting the A. octodonta surface suggesting that the interactions between luminous bacteria and the examined hydrozoan and bryozoan species are highly specific. These interactions might have epidemiological as well as ecological implications because of the opportunistic pathogenicity of luminous Vibrio species for marine organisms and the wide-distribution of the hydrozoan and bryozoan functioning as carriers. PMID:18437448

Stabili, L; Gravili, C; Tredici, S M; Piraino, S; Talà, A; Boero, F; Alifano, P

2008-11-01

189

Enzymatic characterization of Vibrio alginolyticus strains isolated from bivalves harvested at Venice Lagoon (Italy) and Guanabara Bay (Brazil).  

PubMed

The aquatic ecosystem is the natural habitat of microorganisms including Vibrio and Aeromonas genus which are pathogenic to human and animals. In the present investigation the frequency of these bacteria and the enzymatic characteristics of 34 Vibrio alginolyticus strains isolated from bivalves harvested in Venice Lagoon (Italy) and Guanabara Bay (Brazil) were carried out from November 2003 to February 2004. The mussels' samples were submitted to enrichment in Alkaline Peptone Water (APW) added with 1% of sodium chloride (NaCl) and APW plus 3% NaCl incubated at 37 degrees C for 18-24 h. Following the samples were streaked onto TCBS Agar (Thiossulfate Citrate Bile Sucrose Agar) and the suspected colonies were submitted to biochemical characterization. Also, the Vibrio alginolyticus strains were evaluated to collagenase, elastase and chondroitinase production. The results showed the isolation of 127 microorganisms distributed as follows: 105 Vibrio strains such as V. alginolyticus (32.4%), V. harveyi (19%) and V. parahaemolyticus (7.6%), 20 Aeromonas strains and two Plesiomonas shigelloides were the main pathogens isolated. We observed the production of the three enzymes from V. alginolyticus strains considered as the main virulence factors of the bacteria, especially in cases of human dermatological infection. PMID:18813756

Lafisca, Andrea; Pereira, Christiane Soares; Giaccone, Valério; Rodrigues, Dalia dos Prazeres

2008-01-01

190

Facile synthesis of multifunctional multi-walled carbon nanotube for pathogen Vibrio alginolyticus detection in fishery and environmental samples.  

PubMed

Interest in carbon nanotubes for detecting the presence of pathogens arises because of developments in chemical vapor deposition synthesis and progresses in biomolecular modification. Here we reported the facile synthesis of multi-walled carbon nanotubes (MWCNTs), which functioned as immuno-, magnetic, fluorescent sensors in detecting Vibrio alginolyticus (Va). The structures and properties of functionalized MWCNTs were characterized by ultraviolet (UV), Fourier transform infrared spectra (FT-IR), X-ray diffraction (XRD), vibrating sample magnetometer (VSM), magnetic property measurement system (MPMS) and fluorescent spectra (FL). It was found that the functionalized MWCNTs showed: (1) low nonspecific adsorption for antibody-antigen, (2) strong interaction with antibody, and (3) high immune-magnetic activity for pathogenic cells. Further investigations revealed a strong positive linear relationship (R=0.9912) between the fluorescence intensity and the concentration of Va in the range of 9.0×10(2) to 1.5×10(6)cfumL(-1). Moreover, the relative standard deviation for 11 replicate detections of 1.0×10(4)cfumL(-1) Va was 2.4%, and no cross-reaction with the other four strains was found, indicating a good specificity for Va detection. These results demonstrated the remarkable advantages of the multifunctional MWCNTs, which offer great potential for the rapid, sensitive and quantitative detection of Va in fishery and environmental samples. PMID:25059166

Liu, Yue; Hu, Jia; Sun, Jin-Sheng; Li, Yan; Xue, Shu-Xia; Chen, Xiao-Qin; Li, Xiao-Shuang; Du, Gui-Xiang

2014-10-01

191

Comparison of the Pathogenic Potentials of Environmental and Clinical Vibrio parahaemolyticus Strains Indicates a Role for Temperature Regulation in Virulence? †  

PubMed Central

Although the presence of pathogenic Vibrio spp. in estuarine environments of northern New England has been known for some time (C. H. Bartley and L. W. Slanetz, Appl. Microbiol. 21: 965-966, 1971, and K. R. O'Neil, S. H. Jones, and D. J. Grimes, FEMS Microbiol. Lett. 60:163-167, 1990), their virulence and the relative threat they may pose to human health has yet to be evaluated. In this study, the virulence potential of 33 Vibrio parahaemolyticus isolates collected from the Great Bay Estuary of New Hampshire was assessed in comparison to that of clinical strains. The environmental isolates lack thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH), which are encoded by tdh and trh, respectively. Though not hemolytic, they do possess putative virulence factors, such type III secretion system 1, and are highly cytotoxic to human gastrointestinal cells. The expression of known and putative virulence-associated traits, including hemolysin, protease, motility, biofilm formation, and cytotoxicity, by clinical reference isolates correlated with increased temperature from 28°C to 37°C. In contrast, the environmental isolates did not induce their putative virulence-associated traits in response to a temperature of 37°C. We further identified a significant correlation between hemolytic activity and growth phase among clinical strains, whereby hemolysin production decreases with increasing cell density. The introduction of a tdh::gfp promoter fusion into the environmental strains revealed that they regulate this virulence-associated gene appropriately in response to temperature, indicating that their existing regulatory mechanisms are primed to manage newly acquired virulence genes.

Mahoney, Jennifer C.; Gerding, Matthew J.; Jones, Stephen H.; Whistler, Cheryl A.

2010-01-01

192

Comparison of the pathogenic potentials of environmental and clinical vibrio parahaemolyticus strains indicates a role for temperature regulation in virulence.  

PubMed

Although the presence of pathogenic Vibrio spp. in estuarine environments of northern New England has been known for some time (C. H. Bartley and L. W. Slanetz, Appl. Microbiol. 21: 965-966, 1971, and K. R. O'Neil, S. H. Jones, and D. J. Grimes, FEMS Microbiol. Lett. 60:163-167, 1990), their virulence and the relative threat they may pose to human health has yet to be evaluated. In this study, the virulence potential of 33 Vibrio parahaemolyticus isolates collected from the Great Bay Estuary of New Hampshire was assessed in comparison to that of clinical strains. The environmental isolates lack thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH), which are encoded by tdh and trh, respectively. Though not hemolytic, they do possess putative virulence factors, such type III secretion system 1, and are highly cytotoxic to human gastrointestinal cells. The expression of known and putative virulence-associated traits, including hemolysin, protease, motility, biofilm formation, and cytotoxicity, by clinical reference isolates correlated with increased temperature from 28°C to 37°C. In contrast, the environmental isolates did not induce their putative virulence-associated traits in response to a temperature of 37°C. We further identified a significant correlation between hemolytic activity and growth phase among clinical strains, whereby hemolysin production decreases with increasing cell density. The introduction of a tdh::gfp promoter fusion into the environmental strains revealed that they regulate this virulence-associated gene appropriately in response to temperature, indicating that their existing regulatory mechanisms are primed to manage newly acquired virulence genes. PMID:20889774

Mahoney, Jennifer C; Gerding, Matthew J; Jones, Stephen H; Whistler, Cheryl A

2010-11-01

193

Use of Representational Difference Analysis To Identify Genomic Differences between Pathogenic Strains of Vibrio cholerae  

PubMed Central

Representational difference analysis (RDA) is a recently developed technique used for amplifying genetic differences between two closely related genomes. We compared RDA and a modified version of RDA to examine genomic differences between the two Vibrio cholerae serogroups that cause epidemic cholera, O1 and O139, and between the two biotypes of the O1 serogroup. With both techniques, we recovered several sequences known to be found only in V. cholerae O139 but absent in its presumed progenitor, V. cholerae O1 El Tor. A greater number of unique fragments were generated in comparing the two V. cholerae O1 biotypes, consistent with the probable greater genetic differences between the two biotypes.

Calia, Kerstin E.; Waldor, Matthew K.; Calderwood, Stephen B.

1998-01-01

194

Assessment of polyaromatic hydrocarbon degradation by potentially pathogenic environmental Vibrio parahaemolyticus isolates from coastal Louisiana, USA.  

PubMed

A presumed Vibrio parahaemolyticus isolate from Chesapeake Bay, Maryland, USA was previously reported to grow on phenanthrene, a polyaromatic hydrocarbon (PAH) found in crude oil. Following the Deepwater Horizon oil spill in the Gulf of Mexico, concerns were raised that PAH-degrading V. parahaemolyticus could increase in abundance, leading to elevated risks of disease derived from shellfish consumption. To assess this possibility, we examined responses to naphthalene and phenanthrene of 17 coastal Louisiana environmental V. parahaemolyticus isolates representing five distinct genotypes. Isolates were obtained immediately after the spill began and after oil had reached the Louisiana coast. None of the isolates grew on or oxidized either substrate and a naphthalene degradation product, 1-naphthol, substantially inhibited growth of some isolates. The use of PAH by V. parahaemolyticus is unusual, and an increase in human health risks due to stimulation of V. parahaemolyticus growth by oil-derived PAH under in situ conditions appears unlikely. PMID:22063191

Smith, Conor B; Johnson, Crystal N; King, Gary M

2012-01-01

195

Overcoming a Defect in Generalized Recombination in the Marine Fish PathogenVibrio anguillarum775: Construction of arecAMutant by Marker Exchange  

Microsoft Academic Search

A defect in generalized recombination has prevented the use of marker exchange for the construction of specific chromosomal mutations in the marinefish pathogenVibrio anguillarum775. Through the use of large segments of homologous DNA, we were successful in overcoming this defect and used marker exchange to construct a recA mutant of V. anguillarum H775-3. A recombinant cosmid carrying the recA gene

JOHN T. SINGER; CHENGHUA MA; ANDKATHERINE J. BOETTCHER

1996-01-01

196

Quorum Sensing Regulatory Cascades Control Vibrio fluvialis Pathogenesis  

PubMed Central

Quorum sensing (QS) is a process by which individual bacteria are able to communicate with one another, thereby enabling the population as a whole to coordinate gene regulation and subsequent phenotypic outcomes. Communication is accomplished through production and detection of small molecules in the extracellular milieu. In many bacteria, particularly Vibrio species, multiple QS systems result in multiple signals, as well as cross talk between systems. In this study, we identify two QS systems in the halophilic enteric pathogen Vibrio fluvialis: one acyl-homoserine lactone (AHL) based and one CAI-1/AI-2 based. We show that a LuxI homolog, VfqI, primarily produces 3-oxo-C10-HSL, which is sensed by a LuxR homolog, VfqR. VfqR-AHL is required to activate vfqI expression and autorepress vfqR expression. In addition, we have shown that similar to that in V. cholerae and V. harveyi, V. fluvialis produces CAI-1 and AI-2 signal molecules to activate the expression of a V. cholerae HapR homolog through LuxO. Although VfqR-AHL does not regulate hapR expression, HapR can repress vfqR transcription. Furthermore, we found that QS in V. fluvialis positively regulates production of two potential virulence factors, an extracellular protease and hemolysin. QS also affects cytotoxic activity against epithelial tissue cultures. These data suggest that V. fluvialis integrates QS regulatory pathways to play important physiological roles in pathogenesis.

Wang, Yunduan; Wang, Hui; Liang, Weili; Hay, Amanda J.; Zhong, Zengtao

2013-01-01

197

An evaluation of chromogenic substrates for characterization of serine protease produced by pathogenic Vibrio alginolyticus.  

PubMed

Four chromogenic substrates for characterizing serine protease of Vibrio alginolyticus were evaluated. The protease activity of bacterial extracellular products, or the fractions of 33 kD protease purified by the AKTA purifier system with various columns, was completely inhibited by ethylenediamine tetra-acetic acid, ethylene glycol-bis(beta-amino-ethyl ether) N,N,N',N'-tetraacetic acid (EGTA), antipain and phenylmethylsulphonyl fluoride (PMSF) using water-soluble substrates (azoalbumin and azocasein). It was only completely inhibited by antipain and PMSF using water-insoluble substrates (azocoll and hide powder azure). The protease activity was not, or only partially, inhibited by 1,10-phenanthroline and sodium dodecyl sulphate (SDS) using all four substrates. Since chelating agents and 1,10-phenanthroline are commonly employed as inhibitors to identify metalloprotease, the two water-soluble substrates may not be appropriate for this purpose, except for using 1,10-phenanthroline as an inhibitor. Chelating agents may be still applicable as inhibitors using water-insoluble substrates and 1,10-phenanthroline is highly recommended in the characterization for metalloprotease to avoid confusion. In the present study, the 33 kD protease was further confirmed as an SDS-resistant serine protease and not a metalloprotease. PMID:10413876

Chen, F R; Liu, P C; Lee, K K

1999-01-01

198

Isolation and characterization of pathogenic Vibrio alginolyticus from diseased cobia Rachycentron canadum.  

PubMed

Outbreaks of serious mortality among cultured juvenile cobia Rachycentron canadum L. (weighing 8-10 g) characterized by lethargy, dark skin and ascites in the peritoneal cavity while some fish possessing damaged eyes occurred in July and August of 2001 in Taiwan. Fifteen motile bacterial strains were isolated from head kidney and/or the ascites on tryptic soy agar supplemented with 1% NaCl (TSA1) and/or thiosulphate citrate bile salt (TCBS) sucrose agar plates during the two outbreaks. All the isolates were characterized and identified as Vibrio alginolyticus on the basis of biochemical characteristics, and comparisons with those of the reference strain V. alginolyticus ATCC 17749. The strain C3c01 (a representative of the 15 similar field isolates), was virulent to the cobia with an LD50 value of 3.28 x 10(4) colony forming units/g fish body weight. All the moribund/dead fish exhibited lethargy, dark skin and ascites in the peritoneal cavity as that observed in natural outbreaks. The same bacteria could be reisolated from kidney and the ascites of fish after bacterial challenge using TSA1 and TCBS plates. The results reveal that V. alginolyticus is an infectious agent of vibriosis in the cobia. PMID:14768024

Liu, Ping-Chung; Lin, Ji-Yang; Hsiao, Pei-Tze; Lee, Kuo-Kau

2004-01-01

199

Identification of multiple sigma54-dependent transcriptional activators in Vibrio cholerae.  

PubMed

In the pathogenic bacterium Vibrio cholerae, the alternate sigma factor sigma54 is required for expression of multiple sets of genes, including an unidentified gene(s) necessary for enhanced colonization within the host. To identify sigma54-dependent transcriptional activators involved in colonization, PCR was performed with V. cholerae chromosomal DNA and degenerate primers, revealing six novel and distinct coding sequences with homology to sigma54-dependent activators. One sequence had high homology to the luxO gene of V. harveyi, which in that organism is involved in quorum sensing. Phenotypes of V. cholerae strains containing mutations in each of the six putative sigma54-dependent activator genes identified one as a probable ntrC homologue. None of the mutant strains exhibited a defect in the ability to colonize infant mice, suggesting the presence of additional sigma54-dependent activators not identified by this technique. PMID:9748465

Klose, K E; Novik, V; Mekalanos, J J

1998-10-01

200

Polysiphonia harveyi, WNC2005-126  

NSDL National Science Digital Library

WNC2005-126, Polysiphonia harveyi J. Bailey, Floating docks at Banks Channel, Wrightville Beach, New Hanover County, NC, 30 Jan 2005, Coll: DW Freshwater & B Stuercke, Det: DW Freshwater & B Stuercke, Poly NC6

Freshwater, Wilson

2008-03-07

201

FtsK-Dependent Dimer Resolution on Multiple Chromosomes in the Pathogen Vibrio cholerae  

PubMed Central

Unlike most bacteria, Vibrio cholerae harbors two distinct, nonhomologous circular chromosomes (chromosome I and II). Many features of chromosome II are plasmid-like, which raised questions concerning its chromosomal nature. Plasmid replication and segregation are generally not coordinated with the bacterial cell cycle, further calling into question the mechanisms ensuring the synchronous management of chromosome I and II. Maintenance of circular replicons requires the resolution of dimers created by homologous recombination events. In Escherichia coli, chromosome dimers are resolved by the addition of a crossover at a specific site, dif, by two tyrosine recombinases, XerC and XerD. The process is coordinated with cell division through the activity of a DNA translocase, FtsK. Many E. coli plasmids also use XerCD for dimer resolution. However, the process is FtsK-independent. The two chromosomes of the V. cholerae N16961 strain carry divergent dimer resolution sites, dif1 and dif2. Here, we show that V. cholerae FtsK controls the addition of a crossover at dif1 and dif2 by a common pair of Xer recombinases. In addition, we show that specific DNA motifs dictate its orientation of translocation, the distribution of these motifs on chromosome I and chromosome II supporting the idea that FtsK translocation serves to bring together the resolution sites carried by a dimer at the time of cell division. Taken together, these results suggest that the same FtsK-dependent mechanism coordinates dimer resolution with cell division for each of the two V. cholerae chromosomes. Chromosome II dimer resolution thus stands as a bona fide chromosomal process.

Val, Marie-Eve; Kennedy, Sean P.; Karoui, Meriem El; Bonne, Laetitia; Chevalier, Fabien; Barre, Francois-Xavier

2008-01-01

202

Identification of Pathogenic Vibrio Species by Multilocus PCR-Electrospray Ionization Mass Spectrometry and Its Application to Aquatic Environments of the Former Soviet Republic of Georgia? †  

PubMed Central

The Ibis T5000 is a novel diagnostic platform that couples PCR and mass spectrometry. In this study, we developed an assay that can identify all known pathogenic Vibrio species and field-tested it using natural water samples from both freshwater lakes and the Georgian coastal zone of the Black Sea. Of the 278 total water samples screened, 9 different Vibrio species were detected, 114 (41%) samples were positive for V. cholerae, and 5 (0.8%) samples were positive for the cholera toxin A gene (ctxA). All ctxA-positive samples were from two freshwater lakes, and no ctxA-positive samples from any of the Black Sea sites were detected.

Whitehouse, Chris A.; Baldwin, Carson; Sampath, Rangarajan; Blyn, Lawrence B.; Melton, Rachael; Li, Feng; Hall, Thomas A.; Harpin, Vanessa; Matthews, Heather; Tediashvili, Marina; Jaiani, Ekaterina; Kokashvili, Tamar; Janelidze, Nino; Grim, Christopher; Colwell, Rita R.; Huq, Anwar

2010-01-01

203

Nutritive Medium for the Differentiation of Vibrio.  

National Technical Information Service (NTIS)

A medium, prepared and tested in lab practice, can be used to determine the capacity of pure vibrio cultures to form hydrogen sulfide, which is of great importance on differentiating pathogenic from nonpathogenic vibrio cultures. The medium consists of th...

P. A. Voloskov Z. S. Konnova M. S. Luchko

1969-01-01

204

Transcriptional Regulation of opaR, qrr2–4 and aphA by the Master Quorum-Sensing Regulator OpaR in Vibrio parahaemolyticus  

PubMed Central

Background Vibrio parahaemolyticus is a leading cause of infectious diarrhea and enterogastritis via the fecal-oral route. V. harveyi is a pathogen of fishes and invertebrates, and has been used as a model for quorum sensing (QS) studies. LuxR is the master QS regulator (MQSR) of V. harveyi, and LuxR-dependent expression of its own gene, qrr2–4 and aphA have been established in V. harveyi. Molecular regulation of target genes by the V. parahaemolyticus MQSR OpaR is still poorly understood. Methodology/Principal Findings The bioinformatics analysis indicated that V. parahaemolyticus OpaR, V. harveyi LuxR, V. vulnificu SmcR, and V. alginolyticus ValR were extremely conserved, and that these four MQSRs appeared to recognize the same conserved cis-acting signals, which was represented by the consensus constructs manifesting as a position frequency matrix and as a 20 bp box, within their target promoters. The MQSR box-like sequences were found within the upstream DNA regions of opaR, qrr2–4 and aphA in V. parahaemolyticus, and the direct transcriptional regulation of these target genes by OpaR were further confirmed by multiple biochemical experiments including primer extension assay, gel mobility shift assay, and DNase I footprinting analysis. Translation and transcription starts, core promoter elements for sigma factor recognition, Shine-Dalgarno sequences for ribosome recognition, and OpaR-binding sites were determined for the five target genes of OpaR, which gave a structural map of the OpaR-dependent promoters. Further computational promoter analysis indicated the above regulatory circuits were shared by several other closely related Vibrios but with slight exceptions. Conclusions/Significance This study gave a comprehensive computational and characterization of the direct transcriptional regulation of five target genes, opaR, qrr2–4 and ahpA, by OpaR in V. parahaemolyticus. These characterized regulatory circuits were conserved in V. harveyi and V. parahaemolyticus.

Tan, Yafang; Guo, Zhaobiao; Yang, Ruifu; Zhou, Dongsheng

2012-01-01

205

Occurrence and distribution of Vibrio spp., Listonella spp., and Clostridium botulinum in the Seto Inland Sea of Japan.  

PubMed Central

The distribution of Vibrio species in samples of surface water, bottom water (water 2 m above the sediment), and sediment from the Seto Inland Sea was studied. A simple technique using a membrane filter and short preenrichment in alkaline peptone water was developed to resuscitate the injured cells, followed by plating them onto TCBS agar. In addition, a survey was conducted to determine the incidence of Clostridium botulinum in sediment samples. Large populations of heterotrophs were found in surface water, whereas large numbers of total vibrios were found in bottom water. In samples from various water sampling regions, high counts of all bacterial populations were found in the inner regions having little exchange of seawater when compared with those of the open region of the inland sea. In the identification of 463 isolates, 23 Vibrio spp. and 2 Listonella spp. were observed. V. harveyi was prevalent among the members of the Vibrio genus. Vibrio species were categorized into six groups; an estimated 20% of these species were in the so-called "pathogenic to humans" group. In addition, a significant proportion of this group was hemolytic and found in the Bisan Seto region. V. vulnificus, V. fluvialis, and V. cholerae non-O1 predominated in the constricted area of the inland sea, which is eutrophic as a result of riverine influence. It was concluded that salinity indirectly governs the distribution of total vibrios and analysis of variance revealed that all bacterial populations were distributed homogeneously and the variance values were found to be significant in some water sampling regions.(ABSTRACT TRUNCATED AT 250 WORDS)

Venkateswaran, K; Nakano, H; Okabe, T; Takayama, K; Matsuda, O; Hashimoto, H

1989-01-01

206

Production and sequence validation of a complete full length ORF collection for the pathogenic bacterium Vibrio cholerae  

PubMed Central

Cholera, an infectious disease with global impact, is caused by pathogenic strains of the bacterium Vibrio cholerae. High-throughput functional proteomics technologies now offer the opportunity to investigate all aspects of the proteome, which has led to an increased demand for comprehensive protein expression clone resources. Genome-scale reagents for cholera would encourage comprehensive analyses of immune responses and systems-wide functional studies that could lead to improved vaccine and therapeutic strategies. Here, we report the production of the FLEXGene clone set for V. cholerae O1 biovar eltor str. N16961: a complete-genome collection of ORF clones. This collection includes 3,761 sequence-verified clones from 3,887 targeted ORFs (97%). The ORFs were captured in a recombinational cloning vector to facilitate high-throughput transfer of ORF inserts into suitable expression vectors. To demonstrate its application, ?15% of the collection was transferred into the relevant expression vector and used to produce a protein microarray by transcribing, translating, and capturing the proteins in situ on the array surface with 92% success. In a second application, a method to screen for protein triggers of Toll-like receptors (TLRs) was developed. We tested in vitro-synthesized proteins for their ability to stimulate TLR5 in A549 cells. This approach appropriately identified FlaC, and previously uncharacterized TLR5 agonist activities. These data suggest that the genome-scale, fully sequenced ORF collection reported here will be useful for high-throughput functional proteomic assays, immune response studies, structure biology, and other applications.

Rolfs, Andreas; Montor, Wagner R.; Yoon, Sang Sun; Hu, Yanhui; Bhullar, Bhupinder; Kelley, Fontina; McCarron, Seamus; Jepson, Daniel A.; Shen, Binghua; Taycher, Elena; Mohr, Stephanie E.; Zuo, Dongmei; Williamson, Janice; Mekalanos, John; LaBaer, Joshua

2008-01-01

207

Luciferase-dependent oxygen consumption by bioluminescent vibrios  

SciTech Connect

Oxygen uptake due to luciferase in two luminous Vibrio species was estimated in vivo by utilizing inhibitors having specificities for luciferase (decanol) and cytochromes (cyanide). Cyanide titration of respiration revealed a component of oxygen uptake less sensitive to cyanide which was completely inhibitable by low concentrations of decanol. From this it was estimated that in vivo luciferase is responsible for less than 12% (Vibrio harveyi) or 20% (Vibrio fischeri) of the total respiration. From these data in vivo bioluminescent quantum yields are estimated to be not lower than 1.7 and 2.6%, respectively.

Makemson, J.C.

1986-02-01

208

Evidence of a new metabolic capacity in an emerging diarrheal pathogen: lessons from the draft genomes of Vibrio fluvialis strains PG41 and I21563  

PubMed Central

Background Vibrio fluvialis is an emerging diarrheal pathogen for which no genome is currently available. In this work, draft genomes of two closely related clinical strains PG41 and I21563 have been explored. Results V. fluvialis strains PG41 and I21563 were sequenced on the Illumina HiSeq 1000 platform to obtain draft genomes of 5.3 Mbp and 4.4 Mbp respectively. Our genome data reveal the presence of genes involved in ethanolamine utilization, which is further experimentally confirmed by growth analysis. Conclusions Combined in silico and growth analysis establish a new metabolic capacity of V. fluvialis to harvest energy from ethanolamine.

2013-01-01

209

A large-scale epidemiological study to identify bacteria pathogenic to Pacific oyster Crassostrea gigas and correlation between virulence and metalloprotease-like activity.  

PubMed

A 4-year bacteriological survey (2003-2007) of four molluscs cultivated in France and faced with mortality episodes was performed by the French shellfish pathology network. The more abundant bacteria isolated during 92 mortality episodes, occurring mainly in Pacific oyster Crassostrea gigas, were identified by genotyping methods. It allowed us both to confirm the representativeness of Vibrio splendidus and Vibrio aestuarianus bacterial strains and to identify both a large number of Vibrio harveyi-related strains mainly detected during 2007 oyster mortality outbreaks and to a lesser extent bacterial strains identified as Shewanella colwelliana. Because metalloprotease has been reported to constitute a virulence factor in a few Vibrio strains pathogenic for C. gigas, several bacterial strains isolated in this study were screened to evaluate their pathogenicity in C. gigas spat by experimental infection and their ability to produce metalloprotease-like activity in the culture supernatant fluids. A high level (84%) of concordant results between azocaseinase activities and virulence of strains was obtained in this study. Because bacterial metalloprotease activities appeared as a common feature of pathogenic bacteria strains associated with mortality events of C. gigas reared in France, this phenotypic test could be useful for the evaluation of virulence in bacterial strains associated with such mortality episodes. PMID:20012275

Saulnier, Denis; De Decker, Sophie; Haffner, Philippe; Cobret, Laetitia; Robert, Maeva; Garcia, Céline

2010-05-01

210

Non-specific immune response of turbot, Scophthalmus maximus (L.), experimentally infected with a pathogenic Vibrio pelagius.  

PubMed

The effect of a pathogenic Vibrio pelagius, isolated during a mass mortality of turbot larvae, on the non-specific immune response of turbot, Scophthalmus maximus (L.), macrophages was studied both in vitro and in vivo. The in vitro treatment of head kidney (HK) macrophages with viable V. pelagius caused a significant inhibition of the chemiluminescence (CL) response in comparison with untreated macrophages, while incubation with heat-killed bacteria did not affect this response. In vivo, the intraperitoneal injection of V. pelagius resulted in a significant inhibition of the CL response in infected fish at days 1 and 4 post-infection compared with the control fish response. The HK macrophage nitric oxide (NO) production was enhanced by in vitro incubation with intermediate doses of viable V. pelagius (5 x 10(3) and 5 x 10(4) bacteria mL(-1)) and higher doses of the heat-killed bacteria (5 x 10(4)-5 x 10(6) bacteria mL(-1)). In both cases, the NO inhibitor N-omega -nitro-L-arginine was capable of down-regulating the specific NO induction caused by incubation with the bacterial treatments. In contrast, incubation with ECPs at higher doses caused a reduction in NO production. In vivo, a significant enhancement in NO production was also observed in macrophage supernatants at day 10 post-infection. Lysozyme concentration in the serum was also significantly increased in the experimentally infected fish at days 4 and 10 post-injection. In addition, viable V. pelagius and its ECPs significantly reduced HK macrophage viability in vitro, whereas no significant differences in viability were observed during the incubation with heat-killed bacteria. As NO production was enhanced in the experimentally infected fish, the inhibitory effect of the NO donor, S-nitroso-acetyl-penicillamine (SNAP), was tested in vitro in a cell-free assay. The results showed that growth of V. pelagius was significantly inhibited using SNAP at a high concentration (1 mM). PMID:12899407

Villamil, L; Figueras, A; Aranguren, R; Novoa, B

2003-06-01

211

Grimontia indica AK16T, sp. nov., Isolated from a Seawater Sample Reports the Presence of Pathogenic Genes Similar to Vibrio Genus  

PubMed Central

Grimontia indica strain AK16T sp. nov. is the type strain of G. indica sp. nov. a new species within the genus Grimontia. This strain, whose genome is described here, was isolated from seawater sample collected from southeast coast of Palk Bay, India. G. indica AK16T is a Gram-negative, facultative aerobic rod shaped bacterium. There are only two other strains in the genus Grimontia one of which, Grimontia hollisae CIP 101886T, is a reported human pathogen isolated from human stool sample while the other, ‘Grimontia marina IMCC5001T’, was isolated from a seawater sample. As compared to the pathogenic strain Grimontia hollisae CIP 101886T, the strain AK16T lacks some genes for pathogenesis like the accessory colonization factors AcfA and AcfD, which are required for the colonization of the bacterium in the host body. While it carries some pathogenesis genes like OmpU, which are related to pathogenesis of Vibrio strains. This suggests that the life cycle of AK16T may include some pathogenic interactions with marine animal(s), or it may be an opportunistic pathogen. Study of the Grimontia genus is important because of the severe pathogenic traits exhibited by a member of the genus with only three species reported in total. The study will provide some vital information which may be useful in future clinical studies on the genus.

Singh, Aditya; Vaidya, Bhumika; Khatri, Indu; Srinivas, T. N. R.; Subramanian, Srikrishna; Korpole, Suresh; Pinnaka, Anil Kumar

2014-01-01

212

Alterations in Hemolymph and Extrapallial Fluid Parameters in the Manila Clam, Ruditapes philippinarum, Challenged with the Pathogen Vibrio tapetis  

Microsoft Academic Search

In a recent study, we demonstrated the presence of defense factors, competent hemocytes and high enzymatic activities (peptidases, hydrolases, lytic, etc.), in the extrapallial fluid, located between the mantle and the shell, of the Manila clam, Ruditapes philippinarum. In Europe, this species is affected by brown ring disease, an epizootic disease caused by the bacterium Vibrio tapetis. The present work

Bassem Allam; Christine Paillard; Michel Auffret

2000-01-01

213

Distribution of Genes for Virulence and Ecological Fitness among Diverse Vibrio cholerae Population in a Cholera Endemic Area: Tracking the Evolution of Pathogenic Strains  

PubMed Central

The pathogenic strains of Vibrio cholerae that cause acute enteric infections in humans are derived from environmental nonpathogenic strains. To track the evolution of pathogenic V. cholerae and identify potential precursors of new pathogenic strains, we analyzed 324 environmental or clinical V. cholerae isolates for the presence of diverse genes involved in virulence or ecological fitness. Of 251 environmental non-O1, non-O139 strains tested, 10 (3.9%) carried the toxin coregulated pilus (TCP) pathogenicity island encoding TCPs, and the CTX prophage encoding cholera toxin, whereas another 10 isolates carried the TCP island alone, and were susceptible to transduction with CTX phage. Most V. cholerae O1 and O139 strains carried these two major virulence determinants, as well as the Vibrio seventh pandemic islands (VSP-1 and VSP-2), whereas 23 (9.1%) non-O1, non-O139 strains carried several VSP island genes, but none carried a complete VSP island. Conversely, 30 (11.9%) non-O1, non-O139 strains carried type III secretion system (TTSS) genes, but none of 63 V. cholerae O1 or O139 strains tested were positive for TTSS. Thus, the distribution of major virulence genes in the non-O1, non-O139 serogroups of V. cholerae is largely different from that of the O1 or O139 serogroups. However, the prevalence of putative accessory virulence genes (mshA, hlyA, and RTX) was similar in all strains, with the mshA being most prevalent (98.8%) followed by RTX genes (96.2%) and hlyA (94.6%), supporting more recent assumptions that these genes imparts increased environmental fitness. Since all pathogenic strains retain these genes, the epidemiological success of the strains presumably depends on their environmental persistence in addition to the ability to produce major virulence factors. Potential precursors of new pathogenic strains would thus require to assemble a combination of genes for both ecological fitness and virulence to attain epidemiological predominance.

Rahman, M. Hasibur; Biswas, Kuntal; Hossain, M. Anwar; Sack, R. Bradley; Mekalanos, John J.

2008-01-01

214

The HlyU Protein Is a Positive Regulator of rtxA1, a Gene Responsible for Cytotoxicity and Virulence in the Human Pathogen Vibrio vulnificus? †  

PubMed Central

Vibrio vulnificus is an opportunistic human pathogen that preferentially infects compromised iron-overloaded patients, causing a fatal primary septicemia with very rapid progress, resulting in a high mortality rate. In this study we determined that the HlyU protein, a virulence factor in V. vulnificus CMCP6, up-regulates the expression of VV20479, a homologue of the Vibrio cholerae RTX (repeats in toxin) toxin gene that we named rtxA1. This gene is part of an operon together with two other open reading frames, VV20481 and VV20480, that encode two predicted proteins, a peptide chain release factor 1 and a hemolysin acyltransferase, respectively. A mutation in rtxA1 not only contributes to the loss of cytotoxic activity but also results in a decrease in virulence, whereas a deletion of VV20481 and VV20480 causes a slight decrease in virulence but with no effect in cytotoxicity. Activation of the expression of the rtxA1 operon by HlyU occurs at the transcription initiation level by binding of the HlyU protein to a region upstream of this operon.

Liu, Moqing; Alice, Alejandro F.; Naka, Hiroaki; Crosa, Jorge H.

2007-01-01

215

Quorum-sensing regulators control virulence gene expression in Vibrio cholerae  

PubMed Central

The production of virulence factors including cholera toxin and the toxin-coregulated pilus in the human pathogen Vibrio cholerae is strongly influenced by environmental conditions. The well-characterized ToxR signal transduction cascade is responsible for sensing and integrating the environmental information and controlling the virulence regulon. We show here that, in addition to the known components of the ToxR signaling circuit, quorum-sensing regulators are involved in regulation of V. cholerae virulence. We focused on the regulators LuxO and HapR because homologues of these two proteins control quorum sensing in the closely related luminous marine bacterium Vibrio harveyi. Using an infant mouse model, we found that a luxO mutant is severely defective in colonization of the small intestine. Gene arrays were used to profile transcription in the V. cholerae wild type and the luxO mutant. These studies revealed that the ToxR regulon is repressed in the luxO mutant, and that this effect is mediated by another negative regulator, HapR. We show that LuxO represses hapR expression early in log-phase growth, and constitutive expression of hapR blocks ToxR-regulon expression. Additionally, LuxO and HapR regulate a variety of other cellular processes including motility, protease production, and biofilm formation. Together these data suggest a role for quorum sensing in modulating expression of blocks of virulence genes in a reciprocal fashion in vivo.

Zhu, Jun; Miller, Melissa B.; Vance, Russell E.; Dziejman, Michelle; Bassler, Bonnie L.; Mekalanos, John J.

2002-01-01

216

Production and characterization of a novel monoclonal antibody against Vibrio parahaemolyticus F 0F 1 ATP synthase's delta subunit and its application for rapid identification of the pathogen  

Microsoft Academic Search

We raised monoclonal antibodies (MAbs) against Vibrio parahaemolyticus cell extracts. One of the MAbs, designated MAb-VP34, reacted in enzyme-linked immunosorbent assays (ELISAs) with 140 V. parahaemolyticus strains, regardless of serotype or origin. MAb-VP34 did not detectably react with 96 strains belonging to 27 other Vibrio species (except for Vibrio natriegens) or with 29 non-Vibrio species. These results show that MAb-VP34

Junko Sakata; Kentaro Kawatsu; Tadashi Iwasaki; Katsuhiro Tanaka; Shigeo Takenaka; Yuko Kumeda; Hiroshi Kodama

217

[Vibrio infections from food and sea water. Introducing the "VibrioNet"].  

PubMed

Vibrio is a genus of bacteria present in surface and coastal waters as well as in marine organisms worldwide. In many countries, pathogenic Vibrio species are a main cause of bacterial diarrhea, which may result from comsumption of contaminated seafood and fish products or from drinking contaminated water. Vibrio infections may also gain in importance in our regions due to global warming and the increase in the world trade of seafood. The research network "VibrioNet" studies pathogenic Vibrios in the marine environment and in seafood consumed by humans as a potential, new emerging zoonotic agent. An assessment of the risk arising from pathogenic non-cholera-vibrios in central Europe is the target of a multidisciplinary research effort. The research network will be strengthened by cooperations with international partners from countries in which Vibrio infections play a major role (Bangladesh, Chile, India, Thailand, and Vietnam). PMID:22015796

Alter, T; Appel, B; Bartelt, E; Dieckmann, R; Eichhorn, C; Erler, R; Frank, C; Gerdts, G; Gunzer, F; Hühn, S; Neifer, J; Oberheitmann, B; Strauch, E

2011-11-01

218

Seasonal Abundance of Total and Pathogenic Vibrio parahaemolyticus Isolated from American Oysters Harvested in the Mandinga Lagoon System, Veracruz, Mexico: Implications for Food Safety.  

PubMed

The abundance of total and pathogenic Vibrio parahaemolyticus (Vp) strains in American oysters (Crassostrea virginica) harvested in two different harvest sites from the Mandinga lagoon System was evaluated monthly for 1 year (January through December 2012). Frequencies of species-specific genes and pathogenic genes exhibited a seasonal distribution. The annual occurrence of Vp with the species-specific tlh gene (tlh(+)) was significantly higher during the winter windy season (32.50%) and spring dry season (15.0%), with the highest densities observed during spring dry season at 283.50 most probable number (MPN)/g (lagoon bank A, near human settlements), indicating the highest risk of infection during warmer months. Pathogenic Vp tlh(+)/tdh(+) frequency was significantly higher during the winter windy and the spring dry seasons at 22.50 and 10.00%, respectively, with highest densities of 16.22 and 41.05 MPN/g (bank A), respectively. The tlh/trh and tdh/trh gene combinations were also found in Vp isolates during the spring dry season at 1.25 and 1.3%, respectively, with densities of 1.79 and 0.4 MPN/g (bank A), respectively. The orf8 genes were detected during the winter windy season (1.25%) with highest densities of 5.96 MPN/g (bank A) and 3.21 MPN/g (bank B, near mangrove islands and a heron nesting area). Densities of Vp tdh(+) were correlated (R(2) = 0.245, P < 0.015) with those of Vp orf8(+). The seasonal dynamics of Vp harboring pathogenic genes varied with seasonal changes, with very high proportions of Vp tdh(+) and Vp orf8(+) isolates in the winter windy season at 46.2 and 17.0%, respectively, which suggests that environmental factors may differentially affect the abundance of pathogenic subpopulations. Although all densities of total Vp (Vp tlh(+)) were lower than 10(4) MPN/g, thus complying with Mexican regulations, the presence of pathogenic strains is a public health concern. Our results suggest that total Vp densities may not be appropriate for assessing oyster contamination and predicting the risk of infection. Evaluation of the presence of pathogenic strains would be a better approach to protecting public health. PMID:24988011

Flores-Primo, Argel; Pardío-Sedas, Violeta; Lizárraga-Partida, Leonardo; López-Hernández, Karla; Uscanga-Serrano, Roxana; Flores-Hernández, Reyna

2014-07-01

219

Marine Bacillus spp. associated with the egg capsule of Concholepas concholepas (common name "loco") have an inhibitory activity toward the pathogen Vibrio parahaemolyticus.  

PubMed

The pandemic bacterium Vibrio parahaemolyticus, isolated from seawater, sediment, and marine organisms, is responsible for gastroenteric illnesses in humans and also cause diseases in aquaculture industry in Chile and other countries around the world. In this study, bacterial flora with inhibitory activity against pathogenic V. parahaemolyticus were collected from egg capsules of Concholepas concholepas and evaluated. The 16S rRNA fragment was sequenced from each isolated strain to determine its identity using the GenBank database. A phylogenetic analysis was made, and tests for the productions of antibacterial substance were performed using the double-layer method. Forty-five morphotypes of bacterial colonies were isolated, 8 of which presented an inhibitory effect on the growth of V. parahaemolyticus. 16S rRNA sequence and phylogenetic analysis show that these strains constitute taxa that are phylogenetically related to the Bacillus genus and are probably sister species or strains of the species Bacillus pumilus, Bacillus licheniform, or Bacillus sp. It is important to determine the nature of the antibacterial substance to evaluate their potential for use against the pathogen species V. parahaemolyticus. PMID:20454959

Leyton, Yanett; Riquelme, Carlos

2010-10-01

220

Isolation of the pathogen Vibrio tapetis and defense parameters in brown ring diseased Manila clams Ruditapes philippinarum cultivated in England.  

PubMed

The Manila clam Ruditapes philippinarum was introduced for aquacultural purposes to Europe in the 1970s. In 1987, brown ring disease (BRD), caused by Vibrio tapetis, appeared in clams cultivated in Brouënou (Finistère, France) and later became increasingly widespread and was reported in cultivated and wild clams existing on the Atlantic coasts of France and Spain. The present study reports, for the first time, the presence of BRD in clams cultivated in England. The etiologic bacterium was isolated and identified using bacteriological and serological techniques. The defence response of affected clams was also studied and significant changes in the hematological and biochemical characteristics of hemolymph and extrapallial fluids were demonstrated. Significant mobilization of hemocytes toward the extrapallial fluids, in contact with the main site of infection (mantle-periostracal lamina area), was observed, suggesting a role for these pseudo-internal compartments in the preservation of clam health. PMID:10918978

Allam, B; Paillard, C; Howard, A; Le Pennec, M

2000-06-19

221

Crystal Structure of SmcR, a Quorum-sensing Master Regulator of Vibrio vulnificus, Provides Insight into Its Regulation of Transcription*  

PubMed Central

Quorum sensing has been implicated as an important global regulatory system controlling the expression of numerous virulence factors in bacterial pathogens. SmcR, a homologue of Vibrio harveyi LuxR, has been proposed as a quorum-sensing master regulator of Vibrio vulnificus, an opportunistic human pathogen. Previous studies demonstrated that SmcR is essential for the survival and pathogenesis of V. vulnificus, indicating that inhibiting SmcR is an attractive approach to combat infections by the bacteria. Here, we determined the crystal structure of SmcR at 2.1 Å resolution. The protein structure reveals a typical TetR superfamily fold consisting of an N-terminal DNA binding domain and a C-terminal dimerization domain. In vivo and in vitro functional analysis of the dimerization domain suggested that dimerization of SmcR is vital for its biological regulatory function. The N-terminal DNA recognition and binding residues were assigned based on the protein structure and the results of in vivo and in vitro mutagenesis experiments. Furthermore, protein-DNA interaction experiments suggested that SmcR may have a sophisticated mechanism that enables the protein to recognize each of its many target operators with different affinities.

Kim, Yoonjeong; Kim, Byoung Sik; Park, Yu Jin; Choi, Won-Chan; Hwang, Jungwon; Kang, Beom Sik; Oh, Tae-Kwang; Choi, Sang Ho; Kim, Myung Hee

2010-01-01

222

Development of a Multiplex Real-Time PCR Assay with an Internal Amplification Control for the Detection of Total and Pathogenic Vibrio parahaemolyticus Bacteria in Oysters?  

PubMed Central

Vibrio parahaemolyticus is an estuarine bacterium that is the leading cause of shellfish-associated cases of bacterial gastroenteritis in the United States. Our laboratory developed a real-time multiplex PCR assay for the simultaneous detection of the thermolabile hemolysin (tlh), thermostable direct hemolysin (tdh), and thermostable-related hemolysin (trh) genes of V. parahaemolyticus. The tlh gene is a species-specific marker, while the tdh and trh genes are pathogenicity markers. An internal amplification control (IAC) was incorporated to ensure PCR integrity and eliminate false-negative reporting. The assay was tested for specificity against >150 strains representing eight bacterial species. Only V. parahaemolyticus strains possessing the appropriate target genes generated a fluorescent signal, except for a late tdh signal generated by three strains of V. hollisae. The multiplex assay detected <10 CFU/reaction of pathogenic V. parahaemolyticus in the presence of >104 CFU/reaction of total V. parahaemolyticus bacteria. The real-time PCR assay was utilized with a most-probable-number format, and its results were compared to standard V. parahaemolyticus isolation methodology during an environmental survey of Alaskan oysters. The IAC was occasionally inhibited by the oyster matrix, and this usually corresponded to negative results for V. parahaemolyticus targets. V. parahaemolyticus tlh, tdh, and trh were detected in 44, 44, and 52% of the oyster samples, respectively. V. parahaemolyticus was isolated from 33% of the samples, and tdh+ and trh+ strains were isolated from 19 and 26%, respectively. These results demonstrate the utility of the real-time PCR assay in environmental surveys and its possible application to outbreak investigations for the detection of total and pathogenic V. parahaemolyticus.

Nordstrom, Jessica L.; Vickery, Michael C. L.; Blackstone, George M.; Murray, Shelley L.; DePaola, Angelo

2007-01-01

223

Detection of total and pathogenic Vibrio parahaemolyticus in shellfish: comparison of PCR protocols using pR72H or toxR targets with a culture method.  

PubMed

PCR protocols directly applied to enrichment broth cultures were compared with a culture method based on the ISO reference for detection of Vibrio parahaemolyticus in 57 natural bivalve mollusc samples. Comparisons were made on different primer pairs specifically targeting the V. parahaemolyticus-specific toxR gene (Vp-toxR) and pR72H fragment, and also tdh and trh hemolysin genes. The PCR method using these different primer pairs and the culture method were also examined for their limits of detection (LOD). The LODs ranged from 7-24 pg of purified DNA per reaction tube (RT) for primer pair Vp-toxR, but for primer pair pR72H, varied greatly depending on the V. parahaemolyticus strains used (0.7 pg-10.6 ng/RT). The Vp-toxR and pR72H primers allowed the detection of V. parahaemolyticus in 25 and 8 out of the 57 samples, respectively, while only 3 V. parahaemolyticus-positive samples were obtained by the culture method. The effective presence of V. parahaemolyticus in the Vp-toxR-positive samples was confirmed by sequencing the PCR products. The trh and Vp-toxR genes were simultaneously detected in 14% of the samples, which were thus considered as presumptively contaminated with pathogenic V. parahaemolyticus. These results emphasize the need for an efficient survey of both the total and pathogenic V. parahaemolyticus present in seafood in France. The PCR protocol targeting Vp-toxR followed by tdh and trh genes is an efficient and reliable method for the detection of total and presumptively pathogenic V. parahaemolyticus in bivalve molluscs. PMID:19106014

Rosec, Jean-Philippe; Simon, Marie; Causse, Véronique; Boudjemaa, Mireille

2009-02-15

224

The Vibrio parahaemolyticus effector VopC mediates Cdc42-dependent invasion of cultured cells but is not required for pathogenicity in an animal model of infection.  

PubMed

Vibrio parahaemolyticus is a Gram-negative marine bacterium that causes acute gastroenteritis in humans. The virulence of V.?parahaemolyticus is dependent upon a type III secretion system (T3SS2). One effector for T3SS2, VopC, is a homologue of the catalytic domain of cytotoxic necrotizing factor (CNF), and was recently reported to be a Rho family GTPase activator and to be linked to internalization of V.?parahaemolyticus by non-phagocytic cultured cells. Here, we provide direct evidence that VopC deamidates Rac1 and CDC42, but not RhoA, in vivo. Our results alsosuggest that VopC, through its activation of Rac1, contributes to formation of actin stress fibres in infected cells. Invasion of host cells, which occurs at a low frequency, does not seem linked to Rac1 activation, but instead appears to require CDC42. Finally, using an infant rabbit model of V.?parahaemolyticus infection, we show that the virulence of V.?parahaemolyticus is not dependent upon VopC-mediated invasion. Genetic inactivation of VopC did not impair intestinal colonization nor reduce signs of disease, including fluid accumulation, diarrhoea and tissue destruction. Thus, although VopC can promote host cell invasion, such internalization is not a critical step of the disease process, consistent with the traditional view of V.?parahaemolyticus as an extracellular pathogen. PMID:24345190

Okada, Ryu; Zhou, Xiaohui; Hiyoshi, Hirotaka; Matsuda, Shigeaki; Chen, Xiang; Akeda, Yukihiro; Kashimoto, Takashige; Davis, Brigid M; Iida, Tetsuya; Waldor, Matthew K; Kodama, Toshio

2014-06-01

225

Overcoming a defect in generalized recombination in the marine fish pathogen Vibrio anguillarum 775: construction of a recA mutant by marker exchange.  

PubMed

A defect in generalized recombination has prevented the use of marker exchange for the construction of specific chromosomal mutations in the marine fish pathogen Vibrio anguillarum 775. Through the use of large segments of homologous DNA, we were successful in overcoming this defect and used marker exchange to construct a recA mutant of V. anguillarum H775-3. A recombinant cosmid carrying the recA gene of V. anguillarum 775 in the center of a 25-kb cloned DNA insert was isolated by complementation of methyl methanesulfonate (MMS) sensitivity in Escherichia coli HB101. The recA gene was inactivated by inserting a kanamycin resistance gene into recA, and the mutant gene was subsequently introduced into V. anguillarum H775-3 by conjugal mobilization. Isolation of recombinants between cosmid-borne recA::kan sequences and chromosomal DNA was facilitated by the introduction of an incompatible plasmid, and Southern hybridization was used to verify the presence of recA::kan in the chromosomal DNA of the recA mutant. V. anguillarum carrying recA::kan was considerably more sensitive to UV radiation and to MMS than was its parent, and near wild-type levels of resistance to MMS and UV light were restored by introduction of cloned recA genes from both E. coli and V. anguillarum. These results indicate that recA is required for DNA repair in V. anguillarum and demonstrate the utility of this modified marker exchange technique for the construction of mutations in this economically important fish pathogen. PMID:8837428

Singer, J T; Ma, C; Boettcher, K J

1996-10-01

226

The Entner-Doudoroff pathway is obligatory for gluconate utilization and contributes to the pathogenicity of Vibrio cholerae.  

PubMed

The Entner-Doudoroff (ED) pathway has recently been shown to play an important role in sugar catabolism for many organisms although very little information is available on the functionality of this pathway in Vibrio cholerae, the causative agent of cholera. In this study, activation of the genes edd and eda, encoding 6-phosphogluconate dehydratase and 2-keto-3-deoxy-6-phosphogluconate aldolase, was used as a marker of a functional ED pathway in V. cholerae. Transcriptional activation analyses and gene silencing experiments with cells grown in sugar-supplemented M9 medium demonstrated that the ED pathway is functional in V. cholerae and is obligatory for gluconate catabolism. Importantly, selective activation of the ED pathway led to concurrent elevation of transcripts of prime virulence genes (ctxA and tcpA) and their regulator (toxT). Further, lowering of these transcript levels and cholera toxin production in vitro by an ED pathway-defective mutant (strain N16961 with a ?edd mutation [?edd(N16961) strain]) suggested the importance of this pathway in regulating V. cholerae virulence. The in vivo relevance of these data was established as the mutant failed to colonize in suckling mice intestine or to induce fluid accumulation in ligated rabbit ileal loops. Activation of the ED pathway in V. cholerae was shown to inhibit biofilm formation in vitro that could be reversed in the mutant. As further support for these results, comparative transcriptome analysis with cells grown in the presence of glucose or gluconate revealed that a functional ED pathway led to activation of a subset of previously reported in vivo expressed genes. All of these results suggest the importance of the ED pathway in V. cholerae pathogenesis. PMID:22544275

Patra, Tapas; Koley, Hemanta; Ramamurthy, Thandavarayan; Ghose, Asoke C; Nandy, Ranjan K

2012-07-01

227

Improvement of innate immune responses and defense activity in tiger shrimp (Penaeus monodon Fab.) by intramuscular administration of the outer membrane protein Vibrio alginolyticus.  

PubMed

The Outer Membrane Protein (OMP) of Vibrio alginolyticus cell wall was administered intramuscularly (IM) to the tiger shrimp (Penaeus monodon Fab.) at 10, 20, 30 ?g/kg bw. After 14 days infection, the tiger shrimps were challenged with 10(7) bacterial density of Vibrio harveyi for 24 hours. The total haemocyte count (THC), differential haemocyte count (DHC) and amount of total protein plasma (TPP), superoxide dismutase and protease enzyme activity were monitored. The results showed that intramuscular administration of OMP enhanced an immunomodulatory effect and protection against V. harveyi. The beneficial effect of OMP on the tiger shrimp is dose-dependent and OMP-20 ?g/kg bw is an optimal dose after two times of boosters for 14 days against V. harveyi infection. PMID:24058892

Maftuch; Prasetio, E; Sudianto, A; Rozik, M; Nurdiyani, R; Sanusi, E; Nursyam, H; Fariedah, F; Marsoedi; Murachman

2013-01-01

228

Cinnamaldehyde and cinnamaldehyde derivatives reduce virulence in Vibrio spp. by decreasing the DNA-binding activity of the quorum sensing response regulator LuxR  

Microsoft Academic Search

BACKGROUND: To date, only few compounds targeting the AI-2 based quorum sensing (QS) system are known. In the present study, we screened cinnamaldehyde and substituted cinnamaldehydes for their ability to interfere with AI-2 based QS. The mechanism of QS inhibition was elucidated by measuring the effect on bioluminescence in several Vibrio harveyi mutants. We also studied in vitro the ability

Gilles Brackman; Tom Defoirdt; Carol Miyamoto; Peter Bossier; Serge Van Calenbergh; Hans Nelis; Tom Coenye

2008-01-01

229

Enhancement of UV Light Sensitivity of a Vibrio parahaemolyticus O3:K6 Pandemic Strain Due to Natural Lysogenization by a Telomeric Phage?  

PubMed Central

The Vibrio parahaemolyticus O3:K6 pandemic clonal strain was first observed in southern Chile in 2004 and has since caused approximately 8,000 seafood-related diarrhea cases in this region. The massive proliferation of the original clonal population offers a unique opportunity to study the evolution of a bacterial pathogen in its natural environment by detection and characterization of emerging bacterial variants. Here, we describe a group of pandemic variants characterized by the presence of a 42-kb extrachromosomal DNA that can be recovered by alkaline extraction. Upon treatment with mitomycin C, these variants lyse with production of a myovirus containing DNA of equal size to the plasmid but which cannot be recovered by alkaline extraction. Plasmid and phage DNAs show similar restriction patterns corresponding to enzyme sites in a circular permutation. Sequenced regions showed 81 to 99% nucleotide similarity to bacteriophage VHML of Vibrio harveyi. Altogether these observations indicate that the 42-kb plasmid corresponds to a prophage, consisting of a linear DNA with terminal hairpins of a telomeric temperate phage with a linear genome. Bacteria containing the prophage were 7 to 15 times more sensitive to UV radiation, likely due to phage induction by UV irradiation as plasmid curing restored the original sensitivity. The enhanced UV sensitivity could have a significant role in reducing the survival and propagation capability of the V. parahaemolyticus pandemic strain in the ocean.

Zabala, Beatriz; Garcia, Katherine; Espejo, Romilio T.

2009-01-01

230

The TonB3 System in the Human Pathogen Vibrio vulnificus Is under the Control of the Global Regulators Lrp and Cyclic AMP Receptor Protein  

PubMed Central

TonB systems transduce the proton motive force of the cytoplasmic membrane to energize substrate transport through a specific TonB-dependent transporter across the outer membrane. Vibrio vulnificus, an opportunistic marine pathogen that can cause a fatal septicemic disease in humans and eels, possesses three TonB systems. While the TonB1 and TonB2 systems are iron regulated, the TonB3 system is induced when the bacterium grows in human serum. In this work we have determined the essential roles of the leucine-responsive protein (Lrp) and cyclic AMP (cAMP) receptor protein (CRP) in the transcriptional activation of this system. Whereas Lrp shows at least four very distinctive DNA binding regions spread out from position ?59 to ?509, cAMP-CRP binds exclusively in a region centered at position ?122.5 from the start point of the transcription. Our results suggest that both proteins bind simultaneously to the region closer to the RNA polymerase binding site. Importantly, we report that the TonB3 system is induced not only by serum but also during growth in minimal medium with glycerol as the sole carbon source and low concentrations of Casamino Acids. In addition to catabolite repression by glucose, l-leucine acts by inhibiting the binding of Lrp to the promoter region, hence preventing transcription of the TonB3 operon. Thus, this TonB system is under the direct control of two global regulators that can integrate different environmental signals (i.e., glucose starvation and the transition between “feast” and “famine”). These results shed light on new mechanisms of regulation for a TonB system that could be widespread in other organisms.

Crosa, Jorge H.

2012-01-01

231

Acanthamoeba castellanii Promotes the Survival of Vibrio parahaemolyticus?  

PubMed Central

Vibrio parahaemolyticus is a food-borne pathogen that naturally inhabits both marine and estuarine environments. Free-living protozoa exist in similar aquatic environments and function to control bacterial numbers by grazing on free-living bacteria. Protozoa also play an important role in the survival and spread of some pathogenic species of bacteria. We investigated the interaction between the protozoan Acanthamoeba castellanii and the bacterium Vibrio parahaemolyticus. We found that Acanthamoeba castellanii does not prey on Vibrio parahaemolyticus but instead secretes a factor that promotes the survival of Vibrio parahaemolyticus in coculture. These studies suggest that protozoa may provide a survival advantage to an extracellular pathogen in the environment.

Laskowski-Arce, Michelle A.; Orth, Kim

2008-01-01

232

Quantification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae in French Mediterranean coastal lagoons  

PubMed Central

Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae are human pathogens. Little is known about these Vibrio spp. in the coastal lagoons of France. The purpose of this study was to investigate their incidence in water, shellfish and sediment of three French Mediterranean coastal lagoons using the most probable number-polymerase chain reaction (MPN-PCR). In summer, the total number of V. parahaemolyticus in water, sediment, mussels and clams collected from the three lagoons varied from 1 to >1.1 × 103 MPN/l, 0.09 to 1.1 × 103 MPN/ml, 9 to 210 MPN/g and 1.5 to 2.1 MPN/g, respectively. In winter, all samples except mussels contained V. parahaemolyticus, but at very low concentrations. Pathogenic (tdh- or trh2-positive) V. parahaemolyticus were present in water, sediment and shellfish samples collected from these lagoons. The number of V. vulnificus in water, sediment and shellfish samples ranged from 1 to 1.1 × 103 MPN/l, 0.07 to 110 MPN/ml and 0.04 to 15 MPN/g, respectively, during summer. V. vulnificus was not detected during winter. V. cholerae was rarely detected in water and sediment during summer. In summary, results of this study highlight the finding that the three human pathogenic Vibrio spp. are present in the lagoons and constitute a potential public health hazard.

Cantet, Franck; Hervio-Heath, Dominique; Caro, Audrey; Le Mennec, Cecile; Monteil, Caroline; Quemere, Catherine; Jolivet-Gougeon, Anne; Colwell, Rita R.; Monfort, Patrick

2014-01-01

233

Rapid identification and differentiation of Vibrio parahaemolyticus from Vibrio spp. in seafood samples using developed monoclonal antibodies.  

PubMed

Monoclonal antibodies (MAbs) specific to Vibrio parahaemolyticus were successfully generated. According to the specificity of V. parahaemolyticus, MAbs can be classified into 5 groups. The MAbs VP-2D and VP-11H were specific to the O2 and O4 groups of V. parahaemolyticus, respectively. The MAb VP-11B reacted with 11 out of 30 isolates of V. parahaemolyticus used in this study. The MAb VP-516 bound to 27 out of 30 isolates of V. parahaemolyticus and cross reacted with all 10 isolates of V. alginolyticus. The MAb VP-618 demonstrated positive reactivity to 29 out of 30 isolates of V. parahaemolyticus and demonstrated slight cross reactivity to 3 out of 30 isolates of V. harveyi. The sensitivity of the MAbs ranged from 10(8) to 10(7) c.f.u. ml(-1) for V. parahaemolyticus obtained from pure cultures and depended on the group of MAbs. However, the detection capability could be improved to be equivalent to that of the PCR technique following pre-incubation of the samples in alkaline peptone water (APW). Using these MAbs along with MAbs specific to V. alginolyticus (VA-165), V. cholerae (VC-63), V. harveyi (VH-9B and VH-20C) and Vibrio spp. (VC-201) from previous studies, V. parahaemolyticus could be identified and differentiated from Vibrio spp. in various seafood samples including shrimp, green mussels, blood clams and oysters by a simple dot blot immunoassay without the requirement for bacterial isolation or biochemical characterization. PMID:23233121

Prompamorn, Piyanuch; Longyant, Siwaporn; Pengsuk, Chalinan; Sithigorngul, Paisarn; Chaivisuthangkura, Parin

2013-04-01

234

Vibrio fluvialis in Patients with Diarrhea, Kolkata, India  

PubMed Central

We identified 131 strains of Vibrio fluvialis among 400 nonagglutinating Vibrio spp. isolated from patients with diarrhea in Kolkata, India. For 43 patients, V. fluvialis was the sole pathogen identified. Most strains harbored genes encoding hemolysin and metalloprotease; this finding may contribute to understanding of the pathogenicity of V. fluvialis.

Chowdhury, Goutam; Pazhani, Gururaja P.; Dutta, Devarati; Guin, Sucharita; Dutta, Sanjucta; Ghosh, Santanu; Izumiya, Hidemasa; Asakura, Masahiro; Yamasaki, Shinji; Takeda, Yoshifumi; Arakawa, Eiji; Watanabe, Haruo; Mukhopadhyay, Asish K.; Bhattacharya, Mihir K.; Rajendran, K.; Nair, Gopinath Balakrish

2012-01-01

235

Autecology of Vibrio vulnificus and Vibrio parahaemolyticus in tropical waters  

SciTech Connect

Water and shellfish samples collected from estuaries, mangroves, and beaches along the coast of Puerto Rico were examined for Vibrio vulnificus and Vibrio parahaemolyticus. An array of water quality parameters were also measured simultaneous with bacteria sampling. Both species of vibrio were associated with estuary and mangrove locations, and neither was isolated from sandy beaches. Densities of V. vulnificus were negatively correlated with salinity, 10--15 ppt being optimal. V. parahaemolyticus was isolated from sites with salinities between 20 and 35 ppt, the highest densities occurring at 20 ppt. Densities of Vibrio spp. and V. parahaemolyticus for a tropical estuary surpassed those reported for temperate estuaries by several orders of magnitude. Both densities of total Vibrio spp. and V. parahaemolyticus in the water were directly related to densities of fecal coliforms, unlike V. vulnificus. The incidence of ONPG(+) strains among sucrose({minus}) Vibrio spp. served as an indicator of the frequency of V. vulnificus in this group. More than 63% of the V. vulnificus isolated were pathogenic. V. vulnificus and V. parahaemolyticus occupy clearly separate niches within the tropical estuarine-marine ecosystem.

Rivera, S.; Lugo, T.; Hazen, T.C. [Univ. of Puerto Rico, Rio Piedras (Puerto Rico)

1988-12-31

236

Biodiversity of Vibrios  

PubMed Central

Vibrios are ubiquitous and abundant in the aquatic environment. A high abundance of vibrios is also detected in tissues and/or organs of various marine algae and animals, e.g., abalones, bivalves, corals, fish, shrimp, sponges, squid, and zooplankton. Vibrios harbour a wealth of diverse genomes as revealed by different genomic techniques including amplified fragment length polymorphism, multilocus sequence typing, repetetive extragenic palindrome PCR, ribotyping, and whole-genome sequencing. The 74 species of this group are distributed among four different families, i.e., Enterovibrionaceae, Photobacteriaceae, Salinivibrionaceae, and Vibrionaceae. Two new genera, i.e., Enterovibrio norvegicus and Grimontia hollisae, and 20 novel species, i.e., Enterovibrio coralii, Photobacterium eurosenbergii, V. brasiliensis, V. chagasii, V. coralliillyticus, V. crassostreae, V. fortis, V. gallicus, V. hepatarius, V. hispanicus, V. kanaloaei, V. neonatus, V. neptunius, V. pomeroyi, V. pacinii, V. rotiferianus, V. superstes, V. tasmaniensis, V. ezurae, and V. xuii, have been described in the last few years. Comparative genome analyses have already revealed a variety of genomic events, including mutations, chromosomal rearrangements, loss of genes by decay or deletion, and gene acquisitions through duplication or horizontal transfer (e.g., in the acquisition of bacteriophages, pathogenicity islands, and super-integrons), that are probably important driving forces in the evolution and speciation of vibrios. Whole-genome sequencing and comparative genomics through the application of, e.g., microarrays will facilitate the investigation of the gene repertoire at the species level. Based on such new genomic information, the taxonomy and the species concept for vibrios will be reviewed in the next years.

Thompson, Fabiano L.; Iida, Tetsuya; Swings, Jean

2004-01-01

237

Reactive oxygen species generated by a heat shock protein (Hsp) inducing product contributes to Hsp70 production and Hsp70-mediated protective immunity in Artemia franciscana against pathogenic vibrios.  

PubMed

The cytoprotective role of heat shock protein (Hsp70) described in a variety of animal disease models, including vibriosis in farmed aquatic animals, suggests that new protective strategies relying upon the use of compounds that selectively turn on Hsp genes could be developed. The product Tex-OE® (hereafter referred to as Hspi), an extract from the skin of the prickly pear fruit, Opuntia ficus indica, was previously shown to trigger Hsp70 synthesis in a non-stressful situation in a variety of animals, including in a gnotobiotically (germ-free) cultured brine shrimp Artemia franciscana model system. This model system offers great potential for carrying out high-throughput, live-animal screens of compounds that have health benefit effects. By using this model system, we aimed to disclose the underlying cause behind the induction of Hsp70 by Hspi in the shrimp host, and to determine whether the product affects the shrimp in inducing resistance towards pathogenic vibrios. We provide unequivocal evidences indicating that during the pretreatment period with Hspi, there is an initial release of reactive oxygen species (hydrogen peroxide and/or superoxide anion), generated by the added product, in the rearing water and associated with the host. The reactive molecules generated are the triggering factors responsible for causing Hsp70 induction within Artemia. We have also shown that Hspi acts prophylactically at an optimum dose regimen to confer protection against pathogenic vibrios. This salutary effect was associated with upregulation of two important immune genes, prophenoloxidase and transglutaminase of the innate immune system. These findings suggest that inducers of stress protein (e.g. Hsp70) are potentially important modulator of immune responses and might be exploited to confer protection to cultured shrimp against Vibrio infection. PMID:24950414

Baruah, Kartik; Norouzitallab, Parisa; Linayati, Linayati; Sorgeloos, Patrick; Bossier, Peter

2014-10-01

238

Putative Virulence Traits and Pathogenicity of Vibrio cholerae Non-O1, Non-O139 Isolates from Surface Waters in Kolkata, India?  

PubMed Central

Vibrio cholerae non-O1, non-O139 was isolated from natural surface waters from different sites sampled in diarrhea endemic zones in Kolkata, India. Twenty-one of these isolates were randomly selected and included in the characterization. The multiserogroup isolates were compared by their virulence traits with a group of clinical non-O1, non-O139 isolates from the same geographic area. Of the 21 environmental isolates, 6 and 14 strains belonged to Heiberg groups I and II, respectively. Three of the environmental isolates showed resistance to 2,2-diamine-6,7-diisopropylpteridine phosphate. All of the non-O1, non-O139 strains were positive for toxR, and except for one environmental isolate, none of them were positive for tcpA in the PCR assay. None of the isolates were positive for genes encoding cholera toxin (ctxA), heat-stable toxin (est), heat-labile toxin (elt), and Shiga toxin variants (stx) of Escherichia coli. Additionally, except for one environmental isolate (PC32), all were positive for the gene encoding El Tor hemolysin (hly). The culture supernatants of 86% (18 of 21) of the environmental isolates showed a distinct cytotoxic effect on HeLa cells, and some of these strains also produced cell-rounding factor. The lipase, protease, and cell-associated hemagglutination activities and serum resistance properties of the environmental and clinical isolates did not differ much. However, seven environmental isolates exhibited very high hemolytic activities (80 to 100%), while none of the clinical strains belonged to this group. The environmental isolates manifested three adherence patterns, namely, carpet-like, diffuse, and aggregative adherence, and the clinical isolates showed diffuse adherence on HeLa cells. Of the 11 environmental isolates tested for enteropathogenic potential, 8 (73%) induced positive fluid accumulation (?100) in a mouse model, and the reactivities of these isolates were comparable to those of clinical strains of non-O1, non-O139 and toxigenic O139 V. cholerae. Comparison of the counts of the colonized environmental and clinical strains in the mouse intestine showed that the organisms of both groups had similar colonizing efficiencies. These findings indicate the presence of potentially pathogenic V. cholerae non-O1, non-O139 strains in surface waters of the studied sites in Kolkata.

Bag, Prasanta K.; Bhowmik, Poulami; Hajra, Tapas K.; Ramamurthy, T.; Sarkar, Pradipta; Majumder, Mrinmoyee; Chowdhury, Goutam; Das, Suresh C.

2008-01-01

239

Predatory bacteria as natural modulators of Vibrio parahaemolyticus and Vibrio vulnificus in seawater and oysters.  

PubMed

This study shows that naturally occurring Vibrio predatory bacteria (VPB) exert a major role in controlling pathogenic vibrios in seawater and shellfish. The growth and persistence of Vibrio parahaemolyticus and Vibrio vulnificus were assessed in natural seawater and in the Eastern oyster, Crassostrea virginica. The pathogens examined were V. vulnificus strain VV1003, V. parahaemolyticus O1:KUT (KUT stands for K untypeable), and V. parahaemolyticus O3:K6 and corresponding O3:K6 mutants deficient in the toxRS virulence regulatory gene or the rpoS alternative stress response sigma factor gene. Vibrios were selected for streptomycin resistance, which facilitated their enumeration. In natural seawater, oysters bioconcentrated each Vibrio strain for 24 h at 22°C; however, counts rapidly declined to near negligible levels by 72 h. In natural seawater with or without oysters, vibrios decreased more than 3 log units to near negligible levels within 72 h. Neither toxRS nor rpoS had a significant effect on Vibrio levels. In autoclaved seawater, V. parahaemolyticus O3:K6 counts increased 1,000-fold over 72 h. Failure of the vibrios to persist in natural seawater and oysters led to screening of the water samples for VPB on lawns of V. parahaemolyticus O3:K6 host cells. Many VPB, including Bdellovibrio and like organisms (BALOs; Bdellovibrio bacteriovorus and Bacteriovorax stolpii) and Micavibrio aeruginosavorus-like predators, were detected by plaque assay and electron microscopic analysis of plaque-purified isolates from Atlantic, Gulf Coast, and Hawaiian seawater. When V. parahaemolyticus O3:K6 was added to natural seawater containing trace amounts of VPB, Vibrio counts diminished 3 log units to nondetectable levels, while VPB increased 3 log units within 48 h. We propose a new paradigm that VPB are important modulators of pathogenic vibrios in seawater and oysters. PMID:22904049

Richards, Gary P; Fay, Johnna P; Dickens, Keyana A; Parent, Michelle A; Soroka, Douglas S; Boyd, E Fidelma

2012-10-01

240

Phylogenetic analysis of vibrios and related species by means of atpA gene sequences.  

PubMed

We investigated the use of atpA gene sequences as alternative phylogenetic and identification markers for vibrios. A fragment of 1322 bp (corresponding to approximately 88% of the coding region) was analysed in 151 strains of vibrios. The relationships observed were in agreement with the phylogeny inferred from 16S rRNA gene sequence analysis. For instance, the Vibrio cholerae, Vibrio halioticoli, Vibrio harveyi and Vibrio splendidus species groups appeared in the atpA gene phylogenetic analyses, suggesting that these groups may be considered as separate genera within the current Vibrio genus. Overall, atpA gene sequences appeared to be more discriminatory for species differentiation than 16S rRNA gene sequences. 16S rRNA gene sequence similarities above 97% corresponded to atpA gene sequences similarities above 80%. The intraspecies variation in the atpA gene sequence was about 99% sequence similarity. The results showed clearly that atpA gene sequences are a suitable alternative for the identification and phylogenetic study of vibrios. PMID:17978204

Thompson, Cristiane C; Thompson, Fabiano L; Vicente, Ana Carolina P; Swings, Jean

2007-11-01

241

Evidence for the role of horizontal transfer in generating pVT1, a large mosaic conjugative plasmid from the clam pathogen, Vibrio tapetis.  

PubMed

The marine bacterium Vibrio tapetis is the causative agent of the brown ring disease, which affects the clam Ruditapes philippinarum and causes heavy economic losses in North of Europe and in Eastern Asia. Further characterization of V. tapetis isolates showed that all the investigated strains harbored at least one large plasmid. We determined the sequence of the 82,266 bp plasmid pVT1 from the CECT4600(T) reference strain and analyzed its genetic content. pVT1 is a mosaic plasmid closely related to several conjugative plasmids isolated from Vibrio vulnificus strains and was shown to be itself conjugative in Vibrios. In addition, it contains DNA regions that have similarity with several other plasmids from marine bacteria (Vibrio sp., Shewanella sp., Listonella anguillarum and Photobacterium profundum). pVT1 contains a number of mobile elements, including twelve Insertion Sequences or inactivated IS genes and an RS1 phage element related to the CTXphi phage of V. cholerae. The genetic organization of pVT1 underscores an important role of horizontal gene transfer through conjugative plasmid shuffling and transposition events in the acquisition of new genetic resources and in generating the pVT1 modular organization. In addition, pVT1 presents a copy number of 9, relatively high for a conjugative plasmid, and appears to belong to a new type of replicon, which may be specific to Vibrionaceae and Shewanelleacae. PMID:21326607

Erauso, Gaël; Lakhal, Fatma; Bidault-Toffin, Adeline; Le Chevalier, Patrick; Bouloc, Philippe; Paillard, Christine; Jacq, Annick

2011-01-01

242

Evidence for the Role of Horizontal Transfer in Generating pVT1, a Large Mosaic Conjugative Plasmid from the Clam Pathogen, Vibrio tapetis  

Microsoft Academic Search

The marine bacterium Vibrio tapetis is the causative agent of the brown ring disease, which affects the clam Ruditapes philippinarum and causes heavy economic losses in North of Europe and in Eastern Asia. Further characterization of V. tapetis isolates showed that all the investigated strains harbored at least one large plasmid. We determined the sequence of the 82,266 bp plasmid

Gaël Erauso; Fatma Lakhal; Adeline Bidault-Toffin; Patrick Le Chevalier; Philippe Bouloc; Christine Paillard; Annick Jacq; Michael Hensel

2011-01-01

243

Evidence for the Role of Horizontal Transfer in Generating pVT1, a Large Mosaic Conjugative Plasmid from the Clam Pathogen, Vibrio tapetis  

PubMed Central

The marine bacterium Vibrio tapetis is the causative agent of the brown ring disease, which affects the clam Ruditapes philippinarum and causes heavy economic losses in North of Europe and in Eastern Asia. Further characterization of V. tapetis isolates showed that all the investigated strains harbored at least one large plasmid. We determined the sequence of the 82,266 bp plasmid pVT1 from the CECT4600T reference strain and analyzed its genetic content. pVT1 is a mosaic plasmid closely related to several conjugative plasmids isolated from Vibrio vulnificus strains and was shown to be itself conjugative in Vibrios. In addition, it contains DNA regions that have similarity with several other plasmids from marine bacteria (Vibrio sp., Shewanella sp., Listonella anguillarum and Photobacterium profundum). pVT1 contains a number of mobile elements, including twelve Insertion Sequences or inactivated IS genes and an RS1 phage element related to the CTXphi phage of V. cholerae. The genetic organization of pVT1 underscores an important role of horizontal gene transfer through conjugative plasmid shuffling and transposition events in the acquisition of new genetic resources and in generating the pVT1 modular organization. In addition, pVT1 presents a copy number of 9, relatively high for a conjugative plasmid, and appears to belong to a new type of replicon, which may be specific to Vibrionaceae and Shewanelleacae.

Bidault-Toffin, Adeline; Le Chevalier, Patrick; Bouloc, Philippe; Paillard, Christine; Jacq, Annick

2011-01-01

244

Disruption of Cell-to-Cell Signaling Does Not Abolish the Antagonism of Phaeobacter gallaeciensis toward the Fish Pathogen Vibrio anguillarum in Algal Systems  

PubMed Central

Quorum sensing (QS) regulates Phaeobacter gallaeciensis antagonism in broth systems; however, we demonstrate here that QS is not important for antagonism in algal cultures. QS mutants reduced Vibrio anguillarum to the same extent as the wild type. Consequently, a combination of probiotic Phaeobacter and QS inhibitors is a feasible strategy for aquaculture disease control.

Prol Garcia, M. J.; D'Alvise, P. W.

2013-01-01

245

Ethanolamine utilization in Vibrio alginolyticus  

PubMed Central

Abstract Ethanolamine is used as an energy source by phylogenetically diverse bacteria including pathogens, by the concerted action of proteins from the eut-operon. Previous studies have revealed the presence of eutBC genes encoding ethanolamine-ammonia lyase, a key enzyme that breaks ethanolamine into acetaldehyde and ammonia, in about 100 bacterial genomes including members of gamma-proteobacteria. However, ethanolamine utilization has not been reported for any member of the Vibrio genus. Our comparative genomics study reveals the presence of genes that are involved in ethanolamine utilization in several Vibrio species. Using Vibrio alginolyticus as a model system we demonstrate that ethanolamine is better utilized as a nitrogen source than as a carbon source. Reviewers This article was reviewed by Dr. Lakshminarayan Iyer and Dr. Vivek Anantharaman (nominated by Dr. L Aravind).

2012-01-01

246

AphA and LuxR/HapR reciprocally control quorum sensing in vibrios  

PubMed Central

Bacteria cycle between periods when they perform individual behaviors and periods when they perform group behaviors. These transitions are controlled by a cell–cell communication process called quorum sensing, in which extracellular signal molecules, called autoinducers (AIs), are released, accumulate, and are synchronously detected by a group of bacteria. AI detection results in community-wide changes in gene expression, enabling bacteria to collectively execute behaviors such as bioluminescence, biofilm formation, and virulence factor production. In this study, we show that the transcription factor AphA is a master regulator of quorum sensing that operates at low cell density (LCD) in Vibrio harveyi and Vibrio cholerae. In contrast, LuxR (V. harveyi)/HapR (V. cholerae) is the master regulator that operates at high cell density (HCD). At LCD, redundant small noncoding RNAs (sRNAs) activate production of AphA, and AphA and the sRNAs repress production of LuxR/HapR. Conversely, at HCD, LuxR/HapR represses aphA. This network architecture ensures maximal AphA production at LCD and maximal LuxR/HapR production at HCD. Microarray analyses reveal that 300 genes are regulated by AphA at LCD in V. harveyi, a subset of which is also controlled by LuxR. We propose that reciprocal gradients of AphA and LuxR/HapR establish the quorum-sensing LCD and HCD gene expression patterns, respectively.

Rutherford, Steven T.; van Kessel, Julia C.; Shao, Yi; Bassler, Bonnie L.

2011-01-01

247

Cloning and Nucleotide Sequence of the gyrB Gene of Vibrio parahaemolyticus and Its Application in Detection of This Pathogen in Shrimp  

PubMed Central

Because biochemical testing and 16S rRNA sequence analysis have proven inadequate for the differentiation of Vibrio parahaemolyticus from closely related species, we employed the gyrase B gene (gyrB) as a molecular diagnostic probe. The gyrB genes of V. parahaemolyticus and closely related Vibrio alginolyticus were cloned and sequenced. Oligonucleotide PCR primers were designed for the amplification of a 285-bp fragment from within gyrB specific for V. parahaemolyticus. These primers recognized 117 of 117 reference and wild-type V. parahaemolyticus strains, whereas amplification did not occur when 90 strains of 37 other Vibrio species or 60 strains representing 34 different nonvibrio species were tested. In 100-?l PCR mixtures, the lower detection limits were 5 CFU for live cells and 4 pg for purified DNA. The possible application of gyrB primers for the routine identification of V. parahaemolyticus in food was examined. We developed and tested a procedure for the specific detection of the target organism in shrimp consisting of an 18-h preenrichment followed by PCR amplification of the 285-bp V. parahaemolyticus-specific fragment. This method enabled us to detect an initial inoculum of 1.5 CFU of V. parahaemolyticus cells per g of shrimp homogenate. By this approach, we were able to detect V. parahaemolyticus in all of 27 shrimp samples artificially inoculated with this bacterium. We present here a rapid, reliable, and sensitive protocol for the detection of V. parahaemolyticus in shrimp.

Venkateswaran, Kasthuri; Dohmoto, Nobuhiko; Harayama, Shigeaki

1998-01-01

248

Evaluation of bactericidal activity of weakly acidic electrolyzed water (WAEW) against Vibrio vulnificus and Vibrio parahaemolyticus  

Microsoft Academic Search

Vibrio parahaemolyticus and Vibriovulnificus cause severe foodborne illness in humans; thus, to reduce outbreaks of disease, it is clearly important to reduce food contamination by these pathogens. Although electrolyzed oxidizing (EO) water has been reported to exhibit strong bactericidal activities against many pathogens, it has never been tested against V. vulnificus and V. parahaemolyticus. The purpose of this study was

Yaru Quan; Kyoo-Duck Choi; Donghwa Chung; Il-Shik Shin

2010-01-01

249

Genomic and functional analysis of Vibrio phage SIO-2 reveals novel insights into ecology and evolution of marine siphoviruses  

PubMed Central

We report on a genomic and functional analysis of a novel marine siphovirus, the Vibrio phage SIO-2. This phage is lytic for related Vibrio species of great ecological interest including the broadly antagonistic bacterium Vibrio sp. SWAT3 as well as notable members of the Harveyi clade (V. harveyi ATTC BAA-1116 and V. campbellii ATCC 25920). Vibrio phage SIO-2 has a circularly permuted genome of 80,598 bp, which displays unusual features. This genome is larger than that of most known siphoviruses and only 38 of the 116 predicted proteins had homologues in databases. Another divergence is manifest by the origin of core genes, most of which share robust similarities with unrelated viruses and bacteria spanning a wide range of phyla. These core genes are arranged in the same order as in most bacteriophages but they are unusually interspaced at two places with insertions of DNA comprising a high density of uncharacterized genes. The acquisition of these DNA inserts is associated with morphological variation of SIO-2 capsid, which assembles as a large (80 nm) shell with a novel T=12 symmetry. These atypical structural features confer on SIO-2 a remarkable stability to a variety of physical, chemical and environmental factors. Given this high level of functional and genomic novelty, SIO-2 emerges as a model of considerable interest in ecological and evolutionary studies.

Baudoux, A-C.; Hendrix, R.W.; Lander, G.C.; Bailly, X.; Podell, S.; Paillard, C.; Johnson, J.E.; Potter, C.S.; Carragher, B.; Azam, F.

2011-01-01

250

Genomic and functional analysis of Vibrio phage SIO-2 reveals novel insights into ecology and evolution of marine siphoviruses.  

PubMed

We report on a genomic and functional analysis of a novel marine siphovirus, the Vibrio phage SIO-2. This phage is lytic for related Vibrio species of great ecological interest including the broadly antagonistic bacterium Vibrio sp. SWAT3 as well as notable members of the Harveyi clade (V.harveyi ATTC BAA-1116 and V.campbellii ATCC 25920). Vibrio phage SIO-2 has a circularly permuted genome of 80598 bp, which displays unusual features. This genome is larger than that of most known siphoviruses and only 38 of the 116 predicted proteins had homologues in databases. Another divergence is manifest by the origin of core genes, most of which share robust similarities with unrelated viruses and bacteria spanning a wide range of phyla. These core genes are arranged in the same order as in most bacteriophages but they are unusually interspaced at two places with insertions of DNA comprising a high density of uncharacterized genes. The acquisition of these DNA inserts is associated with morphological variation of SIO-2 capsid, which assembles as a large (80 nm) shell with a novel T=12 symmetry. These atypical structural features confer on SIO-2 a remarkable stability to a variety of physical, chemical and environmental factors. Given this high level of functional and genomic novelty, SIO-2 emerges as a model of considerable interest in ecological and evolutionary studies. PMID:22225728

Baudoux, A-C; Hendrix, R W; Lander, G C; Bailly, X; Podell, S; Paillard, C; Johnson, J E; Potter, C S; Carragher, B; Azam, F

2012-08-01

251

Current perspectives on the epidemiology and pathogenesis of clinically significant Vibrio spp.  

PubMed Central

Recent taxonomic advances have now implicated several different Vibrio species as human pathogens. While the most common clinical presentation of Vibrio infection continues to be gastroenteritis, an increasing number of extraintestinal infections are being reported, particularly in immunocompromised individuals. Detection of Vibrio infections requires a good clinical history and the use of appropriate isolation and identification procedures by the laboratory to confirm illnesses attributed to Vibrio species. Except for Vibrio cholerae O1 and Vibrio parahaemolyticus, there is little direct evidence linking the production of a myriad of cell-associated or extracellular factors produced by each species with human disease and pathogenesis. Many questions regarding pathogenic Vibrio species remain unanswered, including their frequency and distribution in environmental specimens (water, shellfish), infective doses, virulence potential of individual isolates, and markers associated with such strains. Images

Janda, J M; Powers, C; Bryant, R G; Abbott, S L

1988-01-01

252

Predicting the Distribution of Vibrio spp. in the Chesapeake Bay: A Vibrio cholerae Case Study  

PubMed Central

Vibrio cholerae, the causative agent of cholera, is a naturally occurring inhabitant of the Chesapeake Bay and serves as a predictor for other clinically important vibrios, including Vibrio parahaemolyticus and Vibrio vulnificus. A system was constructed to predict the likelihood of the presence of V. cholerae in surface waters of the Chesapeake Bay, with the goal to provide forecasts of the occurrence of this and related pathogenic Vibrio spp. Prediction was achieved by driving an available multivariate empirical habitat model estimating the probability of V. cholerae within a range of temperatures and salinities in the Bay, with hydrodynamically generated predictions of ambient temperature and salinity. The experimental predictions provided both an improved understanding of the in situ variability of V. cholerae, including identification of potential hotspots of occurrence, and usefulness as an early warning system. With further development of the system, prediction of the probability of the occurrence of related pathogenic vibrios in the Chesapeake Bay, notably V. parahaemolyticus and V. vulnificus, will be possible, as well as its transport to any geographical location where sufficient relevant data are available.

Magny, Guillaume Constantin de; Long, Wen; Brown, Christopher W.; Hood, Raleigh R.; Huq, Anwar; Murtugudde, Raghu; Colwell, Rita R.

2010-01-01

253

Vibrio parahaemolyticus, enterotoxigenic Escherichia coli, enterohemorrhagic Escherichia coli and Vibrio cholerae  

PubMed Central

This review highlighted the following: (i) pathogenic mechanism of the thermostable direct hemolysin produced by Vibrio parahaemolyticus, especially on its cardiotoxicity, (ii) heat-labile and heat-stable enterotoxins produced by enterotoxigenic Escherichia coli, especially structure–activity relationship of heat-stable enterotoxin, (iii) RNA N-glycosidase activity of Vero toxins (VT1 and VT2) produced by enterohemorrhagic Escherichia coli O157:H7, (iv) discovery of Vibrio cholerae O139, (v) isolation of new variant of Vibrio cholerae O1 El Tor that carries classical ctxB, and production of high concentration of cholera toxin by these strains, and (vi) conversion of viable but nonculturable (VBNC) Vibrio cholerae to culturable state by co-culture with eukaryotic cells.

TAKEDA, Yoshifumi

2011-01-01

254

Complete Genome Sequence of the Marine Fish Pathogen Vibrio anguillarum Harboring the pJM1 Virulence Plasmid and Genomic Comparison with Other Virulent Strains of V. anguillarum and V. ordalii ? †  

PubMed Central

We dissected the complete genome sequence of the O1 serotype strain Vibrio anguillarum 775(pJM1) and determined the draft genomic sequences of plasmidless strains of serotype O1 (strain 96F) and O2? (strain RV22) and V. ordalii. All strains harbor two chromosomes, but 775 also harbors the virulence plasmid pJM1, which carries the anguibactin-producing and cognate transport genes, one of the main virulence factors of V. anguillarum. Genomic analysis identified eight genomic islands in chromosome 1 of V. anguillarum 775(pJM1) and two in chromosome 2. Some of them carried potential virulence genes for the biosynthesis of O antigens, hemolysins, and exonucleases as well as others for sugar transport and metabolism. The majority of genes for essential cell functions and pathogenicity are located on chromosome 1. In contrast, chromosome 2 contains a larger fraction (59%) of hypothetical genes than does chromosome 1 (42%). Chromosome 2 also harbors a superintegron, as well as host “addiction” genes that are typically found on plasmids. Unique distinctive properties include homologues of type III secretion system genes in 96F, homologues of V. cholerae zot and ace toxin genes in RV22, and the biofilm formation syp genes in V. ordalii. Mobile genetic elements, some of them possibly originated in the pJM1 plasmid, were very abundant in 775, resulting in the silencing of specific genes, with only few insertions in the 96F and RV22 chromosomes.

Naka, Hiroaki; Dias, Graciela M.; Thompson, Cristiane C.; Dubay, Christopher; Thompson, Fabiano L.; Crosa, Jorge H.

2011-01-01

255

Connecting type VI secretion, quorum sensing, and c-di-GMP production in fish pathogen Vibrio alginolyticus through phosphatase PppA.  

PubMed

Vibrio alginolyticus, a Gram-negative marine bacterium, has brought about severe economic damage to the mariculture industry by causing vibriosis in various fish species. We are intrigued in the regulation of the pathogenesis in this bacterium. Here, we reported a complex regulatory connection among the newly defined type VI secretion system (T6SS), quorum sensing (QS), and 3',5'-cyclic diguanylic acid (c-di-GMP) signal through the phosphatase PppA encoded in the T6SS gene cluster of V. alginolyticus. Whole-genome transcriptome analysis revealed various regulatory targets of PppA including the T6SS substrate hemolysin coregulated protein (Hcp), quorum sensing regulator LuxR, exotoxin alkaline serine protease (Asp), flagellar proteins, as well as proteins involved in polysaccharide biosynthesis and transport. Western blot analysis showed PppA served as a negative regulator of the expression and secretion of Hcp1. Mutation of pppA resulted in an increased level of the intracellular second messenger c-di-GMP and a decreased expression of the QS regulator LuxR as well as exotoxin Asp. Complementation of intact pppA gene in ?pppA mutant restored the production of c-di-GMP, LuxR, and Asp to the wild-type level. Phenotypic studies suggested that PppA takes part in the modulation of biofilm formation, motility, and cell aggregation. These results demonstrated new roles of PppA in controlling virulence factors and pleiotropic phenotypes and contributed to our understanding of the regulation of pathogenesis in V. alginolyticus. PMID:23021863

Sheng, Lili; Lv, Yuanzhi; Liu, Qin; Wang, Qiyao; Zhang, Yuanxing

2013-03-23

256

Three New Regulators of Swarming in Vibrio parahaemolyticus  

PubMed Central

Movement on surfaces, or swarming motility, is effectively mediated by the lateral flagellar (laf) system in Vibrio parahaemolyticus. Expression of laf is induced by conditions inhibiting rotation of the polar flagellum, which is used for swimming in liquid. However, not all V. parahaemolyticus isolates swarm proficiently. The organism undergoes phase variation between opaque (OP) and translucent (TR) cell types. The OP cell produces copious capsular polysaccharide and swarms poorly, whereas the TR type produces minimal capsule and swarms readily. OP?TR switching is often the result of genetic alterations in the opaR locus. Previously, OpaR, a Vibrio harveyi LuxR homolog, was shown to activate expression of the cpsA locus, encoding capsular polysaccharide biosynthetic genes. Here, we show that OpaR also regulates swarming by repressing laf gene expression. However, in the absence of OpaR, the swarming phenotype remains tightly surface regulated. To further investigate the genetic controls governing swarming, transposon mutagenesis of a TR (?opaR1) strain was performed, and SwrT, a TetR-type regulator, was identified. Loss of swrT, a homolog of V. harveyi luxT, created a profound defect in swarming. This defect could be rescued upon isolation of suppressor mutations that restored swarming. One class of suppressors mapped in swrZ, encoding a GntR-type transcriptional regulator. Overexpression of swrZ repressed laf expression. Using reporter fusions and quantitative reverse transcription-PCR, SwrT was demonstrated to repress swrZ transcription. Thus, we have identified the regulatory link that inhibits swarming of OP strains and have begun to elucidate a regulatory circuit that modulates swarming in TR strains.

Jaques, Sandford; McCarter, Linda L.

2006-01-01

257

The Vibrio cholerae Genome Database  

NSDL National Science Digital Library

The Institute For Genomic Research (TIGR) has placed online the latest versions of the DNA sequence of both chromosomes of the cholera pathogen Vibrio cholerae. The TIGR site offers data access via a hypertext Gene Identification Table, DNA Molecule Information, Gene Name Search, Locus Search, RNA Gene Table, Paralogous Gene Families, a Sequence Search, or by download (FTP). Originally published by Heidelberg et al. in the journal Nature [106:477-483, 2000], further information is available to users via links at the TIGR site.

258

Light-scattering sensor for real-time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate  

PubMed Central

Summary The three most common pathogenic species of Vibrio, Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus, are of major concerns due to increased incidence of water? and seafood?related outbreaks and illness worldwide. Current methods are lengthy and require biochemical and molecular confirmation. A novel label?free forward light?scattering sensor was developed to detect and identify colonies of these three pathogens in real time in the presence of other vibrios in food or water samples. Vibrio colonies grown on agar plates were illuminated by a 635?nm laser beam and scatter?image signatures were acquired using a CCD (charge?coupled device) camera in an automated BARDOT (BActerial Rapid Detection using Optical light?scattering Technology) system. Although a limited number of Vibrio species was tested, each produced a unique light?scattering signature that is consistent from colony to colony. Subsequently a pattern recognition system analysing the collected light?scatter information provided classification in 1?2?min with an accuracy of 99%. The light?scattering signatures were unaffected by subjecting the bacteria to physiological stressors: osmotic imbalance, acid, heat and recovery from a viable but non?culturable state. Furthermore, employing a standard sample enrichment in alkaline peptone water for 6?h followed by plating on selective thiosulphate citrate bile salts sucrose agar at 30°C for ??12?h, the light?scattering sensor successfully detected V.?cholerae, V.?parahaemolyticus and V.?vulnificus present in oyster or water samples in 18?h even in the presence of other vibrios or other bacteria, indicating the suitability of the sensor as a powerful screening tool for pathogens on agar plates.

Huff, Karleigh; Aroonnual, Amornrat; Littlejohn, Amy E. Fleishman; Rajwa, Bartek; Bae, Euiwon; Banada, Padmapriya P.; Patsekin, Valery; Hirleman, E. Daniel; Robinson, J. Paul; Richards, Gary P.; Bhunia, Arun K.

2012-01-01

259

Light-scattering sensor for real-time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate.  

PubMed

The three most common pathogenic species of Vibrio, Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus, are of major concerns due to increased incidence of water- and seafood-related outbreaks and illness worldwide. Current methods are lengthy and require biochemical and molecular confirmation. A novel label-free forward light-scattering sensor was developed to detect and identify colonies of these three pathogens in real time in the presence of other vibrios in food or water samples. Vibrio colonies grown on agar plates were illuminated by a 635?nm laser beam and scatter-image signatures were acquired using a CCD (charge-coupled device) camera in an automated BARDOT (BActerial Rapid Detection using Optical light-scattering Technology) system. Although a limited number of Vibrio species was tested, each produced a unique light-scattering signature that is consistent from colony to colony. Subsequently a pattern recognition system analysing the collected light-scatter information provided classification in 1-2?min with an accuracy of 99%. The light-scattering signatures were unaffected by subjecting the bacteria to physiological stressors: osmotic imbalance, acid, heat and recovery from a viable but non-culturable state. Furthermore, employing a standard sample enrichment in alkaline peptone water for 6?h followed by plating on selective thiosulphate citrate bile salts sucrose agar at 30°C for ??12?h, the light-scattering sensor successfully detected V.?cholerae, V.?parahaemolyticus and V.?vulnificus present in oyster or water samples in 18?h even in the presence of other vibrios or other bacteria, indicating the suitability of the sensor as a powerful screening tool for pathogens on agar plates. PMID:22613192

Huff, Karleigh; Aroonnual, Amornrat; Littlejohn, Amy E Fleishman; Rajwa, Bartek; Bae, Euiwon; Banada, Padmapriya P; Patsekin, Valery; Hirleman, E Daniel; Robinson, J Paul; Richards, Gary P; Bhunia, Arun K

2012-09-01

260

What Genomic Sequence Information Has Revealed About Vibrio Ecology in the Ocean—A Review  

Microsoft Academic Search

To date, the genomes of eight Vibrio strains representing six species and three human pathogens have been fully sequenced and reported. This review compares genomic\\u000a information revealed from these sequencing efforts and what we can infer about Vibrio biology and ecology from this and related genomic information. The focus of the review is on those attributes that allow\\u000a the Vibrios

Darrell Jay Grimes; Crystal N. Johnson; Kevin S. Dillon; Adrienne R. Flowers; Nicholas F. Noriea; Tracy Berutti

2009-01-01

261

Detection of V. harveyi in shrimp postlarvae and hatchery tank water by the Most Probable Number technique with PCR  

Microsoft Academic Search

V. harveyi is the cause of serious disease in the shrimp industry in Thailand during cultivation. In this study, the gyrB gene of V. harveyi NICA, isolated from shrimp in Thailand, was sequenced. A pair of specific primers (A2B3) was designed that allowed amplification of a 363 bp gene fragment of V. harveyi. No cross reaction was detected in 17 other

Sawitree Thaithongnum; Pimonsri Ratanama; Karnchana Weeradechapol; Ampaitip Sukhoom; Varaporn Vuddhakul

2006-01-01

262

Draft Genome Sequence of Vibrio fortis Dalian14 Isolated from Diseased Sea Urchin (Strongylocentrotus intermedius).  

PubMed

Here, we report the draft genome sequence of Vibrio fortis Dalian14 isolated from diseased sea urchin (Strongylocentrotus intermedius) during disease outbreaks in North China. The availability of this genome sequence will facilitate the study of the mechanisms of pathogenicity and evolution of Vibrio species. PMID:24994792

Ding, Jun; Dou, Yan; Wang, Yinan; Chang, Yaqing

2014-01-01

263

Evaluation of a rapid polymerase chain reaction based identification technique for Vibrio cholerae isolates  

Microsoft Academic Search

Rapid and accurate identification of waterborne pathogens, such as Vibrio cholerae, in drinking- water sources is important to enable effective resource management and public health protection. Phenotypic systems currently being used for the identification of Vibrio cholerae isolates are time-consuming and the need exists for the development of suitable molecular techniques that can offer both fast and reliable identification. During

W. J. le Roux; D. Masoabi; C. M. E. de Wet; S. N. Venter

264

Draft Genome Sequence of Vibrio fortis Dalian14 Isolated from Diseased Sea Urchin (Strongylocentrotus intermedius)  

PubMed Central

Here, we report the draft genome sequence of Vibrio fortis Dalian14 isolated from diseased sea urchin (Strongylocentrotus intermedius) during disease outbreaks in North China. The availability of this genome sequence will facilitate the study of the mechanisms of pathogenicity and evolution of Vibrio species.

Ding, Jun; Dou, Yan; Wang, Yinan

2014-01-01

265

Nontoxigenic Vibrio parahaemolyticus Strains Causing Acute Gastroenteritis  

PubMed Central

We investigated the virulence properties of four Vibrio parahaemolyticus strains causing acute gastroenteritis following consumption of indigenous mussels in Italy. The isolated strains were cytotoxic and adhesive but, surprisingly, lacked tdh, trh, and type three secretion system 2 (T3SS2) genes. We emphasize that nontoxigenic V. parahaemolyticus can induce acute gastroenteritis, highlighting the need for more investigation of the pathogenicity of this microorganism.

Leoni, Francesca; Serra, Roberto; Serracca, Laura; Decastelli, Lucia; Rocchegiani, Elena; Masini, Laura; Canonico, Cristina; Talevi, Giulia; Carraturo, Antonio

2012-01-01

266

A homoserine lactone autoinducer regulates virulence of an insect-pathogenic bacterium, Xenorhabdus nematophilus (Enterobacteriaceae).  

PubMed Central

N-beta-Hydroxybutanoyl homoserine lactone (HBHL), the autoinducer of the luminescent system of Vibrio harveyi, has been identified as the first small compound to restore virulence to avirulent mutants of Xenorhabdus nematophilus. HBHL stimulated the level of lipase activity excreted by avirulent X. nematophilus and lowered the phenoloxidase activity in the hemolymph of insects infected with X. nematophilus, parameters that are both associated with insect pathogenesis. Moreover, mortality of the insects infected with avirulent X. nematophilus was restored upon injection with HBHL. Chloroform extraction of medium conditioned with wild-type but not avirulent X. nematophilus led to the isolation of a compound with the same chromatographic mobility as HBHL as well as the ability to stimulate the luminescence of a dim autoinducer-dependent mutant of V. harveyi. Transfer of the V. harveyi lux operon into avirulent and wild-type X. nematophilus generated dim and bright luminescent strains, respectively, which responded to HBHL and an agonist and antagonist in a manner analogous to their effects on the luminescence of dim autoinducer-deficient and bright wild-type strains of V. harveyi, indicating that similar HBHL-dependent regulatory systems exist in these two bacterial species.

Dunphy, G; Miyamoto, C; Meighen, E

1997-01-01

267

The nucleotide sequence of Beneckea harveyi 5S rRNA. [bioluminescent marine bacterium  

NASA Technical Reports Server (NTRS)

The primary sequence of the 5S ribosomal RNA isolated from the free-living bioluminescent marine bacterium Beneckea harveyi is reported and discussed in regard to indications of phylogenetic relationships with the bacteria Escherichia coli and Photobacterium phosphoreum. Sequences were determined for oligonucleotide products generated by digestion with ribonuclease T1, pancreatic ribonuclease and ribonuclease T2. The presence of heterogeneity is indicated for two sites. The B. harveyi sequence can be arranged into the same four helix secondary structures as E. coli and other prokaryotic 5S rRNAs. Examination of the 5S-RNS sequences of the three bacteria indicates that B. harveyi and P. phosphoreum are specifically related and share a common ancestor which diverged from an ancestor of E. coli at a somewhat earlier time, consistent with previous studies.

Luehrsen, K. R.; Fox, G. E.

1981-01-01

268

Identification of Vibrio isolates by a multiplex PCR assay and rpoB sequence determination.  

PubMed

Vibrio, a diverse genus of aquatic bacteria, currently includes 72 species, 12 of which occur in human clinical samples. Of these 12, three species--Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus-account for the majority of Vibrio infections in humans. Rapid and accurate identification of Vibrio species has been problematic because phenotypic characteristics are variable within species and biochemical identification requires 2 or more days to complete. To facilitate the identification of human-pathogenic species, we developed a multiplex PCR that uses species-specific primers to amplify gene regions in four species (V. cholerae, V. parahaemolyticus, V. vulnificus, and V. mimicus). The assay was tested on a sample of 309 Vibrio isolates representing 26 named species (including 12 human pathogens) that had been characterized by biochemical methods. A total of 190 isolates that had been identified as one of the four target species all yielded results consistent with the previous classification. The assay identified an additional four V. parahaemolyticus isolates among the other 119 isolates. Sequence analysis based on rpoB was used to validate the multiplex results for these four isolates, and all clustered with other V. parahaemolyticus sequences. The rpoB sequences for 12 of 15 previously unidentified isolates clustered with other Vibrio species in a phylogenetic analysis, and three isolates appeared to represent unnamed Vibrio species. The PCR assay provides a simple, rapid, and reliable tool for identification of the major Vibrio pathogens in clinical samples, and rpoB sequencing provides an additional identification tool for other species in the genus Vibrio. PMID:17093013

Tarr, Cheryl L; Patel, Jayna S; Puhr, Nancy D; Sowers, Evangeline G; Bopp, Cheryl A; Strockbine, Nancy A

2007-01-01

269

Rapid detection of Vibrio species using liquid microsphere arrays and real-time PCR targeting the ftsZ locus.  

PubMed

The development of rapid and sensitive molecular techniques for the detection of Vibrio species would be useful for the surveillance of sporadic infections and management of major outbreaks. Comparative sequence analysis of the ftsZ gene in the predominant Vibrio species that cause human disease revealed distinct alleles for each examined species, including Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus. Light Upon eXtension (LUX) real-time PCR assays were developed to target these species-specific polymorphisms, and were successful in rapidly differentiating the major pathogenic Vibrio species. Luminex liquid microsphere array technology was used to develop a comprehensive assay capable of simultaneously detecting V. cholerae, V. parahaemolyticus and V. vulnificus. These assays permitted the identification of a presumptive V. parahaemolyticus isolate as Vibrio alginolyticus, which was verified using additional molecular characterization. PMID:17172518

Tracz, Dobryan M; Backhouse, Paul G; Olson, Adam B; McCrea, Joanne K; Walsh, Julie A; Ng, Lai-King; Gilmour, Matthew W

2007-01-01

270

Identification of Vibrio splendidus as a Member of the Planktonic Luminous Bacteria from the Persian Gulf and Kuwait Region with luxA Probes  

PubMed Central

Hybridization probes specific for the luxA genes of four groups of luminous bacteria were used to screen luminous isolates obtained from the Persian Gulf, near Al Khiran, Kuwait Nine of these isolates were identified as Vibrio harveyi, a commonly encountered planktonic isolate, while three others showed no hybridization to any of the four probes (V. harveyi, Vibrio fischeri, Photobacterium phosphoreum, or Photobacterium leiognathi) under high-stringency conditions. Polymerase chain reaction amplification was used to prepare a luxA probe against one of these isolates, K-1, and this probe was screened under high-stringency conditions against a collection of DNAs from luminous bacteria; it was found to hybridize specifically to the DNA of the species Vibrio splendidus. A probe prepared against the type strain of V. splendidus (ATCC 33369) was tested against the collection of luminous bacterial DNA preparations and against the Kuwait isolates and was found to hybridize only against the type strain and the three unidentified Kuwait isolates. Extensive taxonomic analysis by standard methods confirmed the identification of the 13 isolates. Images

Nealson, K. H.; Wimpee, B.; Wimpee, C.

1993-01-01

271

Evaluation of nonisotopic DNA hybridization methods for detection of the tdh gene of vibrio parahaemolyticus.  

PubMed

Production of the thermostable direct hemolysin (TDH) by Vibrio parahaemolyticus is associated with pathogenicity of the organism and is encoded by the tdh gene. The timely resolution of seafood-associated outbreaks requires rapid and accurate detection of pathogenic V. parahaemolyticus. The specificity of alkaline phosphatase- and digoxigenin-labeled tdh gene probes was evaluated against 61 strains of V. parahaemolyticus (including isolates from recent outbreaks involving oysters from the Pacific Northwest, Texas, and New York), 85 strains of other vibrios, and 7 strains of non-vibrio species from clinical and environmental sources. The probes were specific for detection of the V. parahaemolyticus tdh gene. PMID:11131887

McCarthy, S A; DePaola, A; Kaysner, C A; Hill, W E; Cook, D W

2000-12-01

272

Differential metabolic responses of clam Ruditapes philippinarum to Vibrio anguillarum and Vibrio splendidus challenges.  

PubMed

Clam Ruditapes philippinarum is one of the important marine aquaculture species in North China. However, pathogens can often cause diseases and lead to massive mortalities and economic losses of clam. In this work, we compared the metabolic responses induced by Vibrio anguillarum and Vibrio splendidus challenges towards hepatopancreas of clam using NMR-based metabolomics. Metabolic responses suggested that both V. anguillarum and V. splendidus induced disturbances in energy metabolism and osmotic regulation, oxidative and immune stresses with different mechanisms, as indicated by correspondingly differential metabolic biomarkers (e.g., amino acids, ATP, glucose, glycogen, taurine, betaine, choline and hypotaurine) and altered mRNA expression levels of related genes including ATP synthase, ATPase, glutathione peroxidase, heat shock protein 90, defensin and lysozyme. However, V. anguillarum caused more severe oxidative and immune stresses in clam hepatopancreas than V. splendidus. Our results indicated that metabolomics could be used to elucidate the biological effects of pathogens to the marine clam R. philippinarum. PMID:24056279

Liu, Xiaoli; Ji, Chenglong; Zhao, Jianmin; Wu, Huifeng

2013-12-01

273

Oleic Acid Produced by a Marine Vibrio spp. Acts as an Anti-Vibrio parahaemolyticus Agent  

PubMed Central

It is known that some strains of Vibrio parahaemolyticus are responsible for gastroenteric diseases caused by the ingestion of marine organisms contaminated with these bacterial strains. Organic products that show inhibitory activity on the growth of the pathogenic V. parahaemolyticus were extracted from a Vibrio native in the north of Chile. The inhibitory organic products were isolated by reverse phase chromatography and permeation by Sephadex LH20, and were characterized by spectroscopic and spectrometric techniques. The results showed that the prevailing active product is oleic acid, which was compared with standards by gas chromatography and high-performance liquid chromatography (HPLC). These active products might be useful for controlling the proliferation of pathogenic clones of V. parahaemolyticus.

Leyton, Yanett; Borquez, Jorge; Darias, Jose; Cueto, Mercedes; Diaz-Marrero, Ana R.; Riquelme, Carlos

2011-01-01

274

Molecular Characterization of Direct Target Genes and cis-Acting Consensus Recognized by Quorum-Sensing Regulator AphA in Vibrio parahaemolyticus  

PubMed Central

Background AphA is the master quorum-sensing (QS) regulator operating at low cell density in vibrios. Molecular regulation of target genes by AphA has been characterized in Vibrio harveyi and V. cholerae, but it is still poorly understood in V. parahaemolyticus. Methodology/Principal Findings The AphA proteins are extremely conserved in V. parahaemolyticus, Vibrio sp. Ex25, Vibrio sp. EJY3, V. harveyi, V. vulnificus, V. splendidus, V. anguillarum, V. cholerae, and V. furnissii. The above nine AphA orthologs appear to recognize conserved cis-acting DNA signals which can be represented by two consensus constructs, a 20 bp box sequence and a position frequency matrix. V. parahaemolyticus AphA represses the transcription of ahpA, qrr4, and opaR through direct AphA-target promoter DNA association, while it inhibits the qrr2-3 transcription in an indirect manner. Translation and transcription starts, core promoter elements for sigma factor recognition, Shine-Dalgarno sequences for ribosome recognition, and AphA-binding sites (containing corresponding AphA box-like sequences) were determined for the three direct AphA targets ahpA, qrr4, and opaR in V. parahaemolyticus. Conclusions/Significance AphA-mediated repression of ahpA, qrr2-4, and opaR was characterized in V. parahaemolyticus by using multiple biochemical and molecular experiments. The computational promoter analysis indicated the conserved mechanism of transcriptional regulation of QS regulator-encoding genes ahpA, qrr4, and opaR in vibrios.

Sun, Fengjun; Zhang, Yiquan; Wang, Li; Yan, Xiaojuan; Tan, Yafang; Guo, Zhaobiao; Qiu, Jingfu; Yang, Ruifu; Xia, Peiyuan; Zhou, Dongsheng

2012-01-01

275

rbcL sequences reveal multiple cryptic introductions of the Japanese red alga Polysiphonia harveyi  

Microsoft Academic Search

In Europe, the last 20 years have seen a spectacular increase in accidental introductions of marine species, but it has recently been suggested that both the actual number of invaders and their impacts have been seriously underestimated because of the prevalence of sibling species in marine habitats. The red alga Polysiphonia harveyi is regarded as an alien in the British

Lynne McIvor; Christine A. Maggs; Jim Provan; Michael J. Stanhope

2001-01-01

276

Interplay between Cyclic AMP-Cyclic AMP Receptor Protein and Cyclic di-GMP Signaling in Vibrio cholerae Biofilm Formation  

Microsoft Academic Search

Vibrio cholerae is a facultative human pathogen. The ability of V. cholerae to form biofilms is crucial for its survival in aquatic habitats between epidemics and is advantageous for host-to-host transmission during epidemics. Formation of mature biofilms requires the production of extracellular matrix components, includ- ing Vibrio polysaccharide (VPS) and matrix proteins. Biofilm formation is positively controlled by the tran-

Jiunn C. N. Fong; Fitnat H. Yildiz

2008-01-01

277

Draft Genome Sequence of Environmental Vibrio cholerae 2012EL-1759 with Similarities to the V. cholerae O1 Classical Biotype.  

PubMed

Vibrio cholerae 2012EL-1759 is an environmental isolate from Haiti that was recovered in 2012 during a cholera outbreak. The genomic backbone is similar to that of the prototypical V. cholerae O1 classical biotype strain O395, and it carries the Vibrio pathogenicity islands (VPI-1 and VPI-2) and a cholera toxin (CTX) prephage. PMID:25013135

Katz, Lee S; Turnsek, Maryann; Kahler, Amy; Hill, Vincent R; Boyd, E Fidelma; Tarr, Cheryl L

2014-01-01

278

Enumeration of Vibrio parahaemolyticus in the viable but nonculturable state using direct plate counts and recognition of individual gene fluorescence in situ hybridization.  

PubMed

Vibrio parahaemolyticus is a gram-negative, halophilic bacterium indigenous to marine and estuarine environments and it is capable of causing food and water-borne illness in humans. It can also cause disease in marine animals, including cultured species. Currently, culture-based techniques are used for quantification of V. parahaemolyticus in environmental samples; however, these can be misleading as they fail to detect V. parahaemolyticus in a viable but nonculturable (VBNC) state which leads to an underestimation of the population density. In this study, we used a novel fluorescence visualization technique, called recognition of individual gene fluorescence in situ hybridization (RING-FISH), which targets chromosomal DNA for enumeration. A polynucleotide probe labeled with Cyanine 3 (Cy3) was created corresponding to the ubiquitous V. parahaemolyticus gene that codes for thermolabile hemolysin (tlh). When coupled with the Kogure method to distinguish viable from dead cells, RING-FISH probes reliably enumerated total, viable V. parahaemolyticus. The probe was tested for sensitivity and specificity against a pure culture of tlh(+), tdh(-), trh(-)V. parahaemolyticus, pure cultures of Vibrio vulnificus, Vibrio harveyi, Vibrio alginolyticus and Vibrio fischeri, and a mixed environmental sample. This research will provide additional tools for a better understanding of the risk these environmental organisms pose to human health. PMID:21329738

Griffitt, Kimberly J; Noriea, Nicholas F; Johnson, Crystal N; Grimes, D Jay

2011-05-01

279

Substrate Specificity and Function of the Pheromone Receptor AinR in Vibrio fischeri ES114  

PubMed Central

Two distinct but interrelated pheromone-signaling systems, LuxI/LuxR and AinS/AinR, positively control bioluminescence in Vibrio fischeri. Although each system generates an acyl-homoserine lactone (AHL) signal, the protein sequences of LuxI/LuxR and AinS/AinR are unrelated. AinS and LuxI generate the pheromones N-octanoyl-AHL (C8-AHL) and N-3-oxo-hexanoyl-AHL (3OC6-AHL), respectively. LuxR is a transcriptional activator that responds to 3OC6-AHL, and to a lesser extent to C8-AHL. AinR is hypothesized to respond to C8-AHL and, based on homology to Vibrio harveyi LuxN, to mediate the repression of a Qrr regulatory RNA. However, a ?ainR mutation decreased luminescence, which was not predicted based on V. harveyi LuxN, raising the possibility of a distinct regulatory mechanism for AinR. Here we show that ainR can complement a luxN mutant, suggesting functional similarity. Moreover, in V. fischeri, we observed ainR-dependent repression of a Pqrr-lacZ transcriptional reporter in the presence of C8-AHL, consistent with its hypothesized regulatory role. The system appears quite sensitive, with a half-maximal effect on a Pqrr reporter at 140 pM C8-AHL. Several other AHLs with substituted and unsubstituted acyl chains between 6 and 10 carbons also displayed an AinR-dependent effect on Pqrr-lacZ; however, AHLs with acyl chains of four carbons or 12 or more carbons lacked activity. Interestingly, 3OC6-AHL also affected expression from the qrr promoter, but this effect was largely luxR dependent, indicating a previously unknown connection between these systems. Finally, we propose a preliminary explanation for the unexpected luminescence phenotype of the ?ainR mutant.

Kimbrough, John H.

2013-01-01

280

Substrate specificity and function of the pheromone receptor AinR in Vibrio fischeri ES114.  

PubMed

Two distinct but interrelated pheromone-signaling systems, LuxI/LuxR and AinS/AinR, positively control bioluminescence in Vibrio fischeri. Although each system generates an acyl-homoserine lactone (AHL) signal, the protein sequences of LuxI/LuxR and AinS/AinR are unrelated. AinS and LuxI generate the pheromones N-octanoyl-AHL (C8-AHL) and N-3-oxo-hexanoyl-AHL (3OC6-AHL), respectively. LuxR is a transcriptional activator that responds to 3OC6-AHL, and to a lesser extent to C8-AHL. AinR is hypothesized to respond to C8-AHL and, based on homology to Vibrio harveyi LuxN, to mediate the repression of a Qrr regulatory RNA. However, a ?ainR mutation decreased luminescence, which was not predicted based on V. harveyi LuxN, raising the possibility of a distinct regulatory mechanism for AinR. Here we show that ainR can complement a luxN mutant, suggesting functional similarity. Moreover, in V. fischeri, we observed ainR-dependent repression of a Pqrr-lacZ transcriptional reporter in the presence of C8-AHL, consistent with its hypothesized regulatory role. The system appears quite sensitive, with a half-maximal effect on a Pqrr reporter at 140 pM C8-AHL. Several other AHLs with substituted and unsubstituted acyl chains between 6 and 10 carbons also displayed an AinR-dependent effect on Pqrr-lacZ; however, AHLs with acyl chains of four carbons or 12 or more carbons lacked activity. Interestingly, 3OC6-AHL also affected expression from the qrr promoter, but this effect was largely luxR dependent, indicating a previously unknown connection between these systems. Finally, we propose a preliminary explanation for the unexpected luminescence phenotype of the ?ainR mutant. PMID:24056099

Kimbrough, John H; Stabb, Eric V

2013-11-01

281

Complete Genome Sequence of Vibrio anguillarum M3, a Serotype O1 Strain Isolated from Japanese Flounder in China  

PubMed Central

Vibrio anguillarum is an important bacterial pathogen that causes vibriosis in marine fish. We present the complete genome sequence of V. anguillarum M3, a serotype O1 clinical strain isolated from Japanese flounder (Paralichthys olivaceus) in Shandong, China.

Li, Guiyang; Li, Jie; Xiao, Peng; Hao, Bin

2013-01-01

282

Genotype to phenotype: identification of diagnostic vibrio phenotypes using whole genome sequences.  

PubMed

Vibrios are ubiquitous in the aquatic environment and can be found in association with animal or plant hosts. The range of ecological relationships includes pathogenic and mutualistic associations. To gain a better understanding of the ecology of these microbes, it is important to determine their phenotypic features. However, the traditional phenotypic characterization of vibrios has been expensive, time-consuming and restricted in scope to a limited number of features. In addition, most of the commercial systems applied for phenotypic characterization cannot characterize the broad spectrum of environmental strains. A reliable and possible alternative is to obtain phenotypic information directly from whole genome sequences. The aim of the present study was to evaluate the usefulness of whole genome sequences as a source of phenotypic information. We performed a comparison of the vibrio phenotypes obtained from the literature with the phenotypes obtained from whole genome sequences. We observed a significant correlation between the previously published phenotypic data and the phenotypic data retrieved from whole genome sequences of vibrios. Analysis of 26 vibrio genomes revealed that all genes coding for the specific proteins involved in the metabolic pathways responsible for positive phenotypes of the 14 diagnostic features (Voges-Proskauer reaction, indole production, arginine dihydrolase, ornithine decarboxylase, utilization of myo-inositol, sucrose and L-leucine, and fermentation of D-mannitol, D-sorbitol, L-arabinose, trehalose, cellobiose, D-mannose and D-galactose) were found in the majority of the vibrios genomes. Vibrio species that were negative for a given phenotype revealed the absence of all or several genes involved in the respective biochemical pathways, indicating the utility of this approach to characterize the phenotypes of vibrios. The absence of the global regulation and regulatory proteins in the Vibrio parahaemolyticus genome indicated a non-vibrio phenotype. Whole genome sequences represent an important source for the phenotypic identification of vibrios. PMID:24505074

Amaral, Gilda Rose S; Dias, Graciela M; Wellington-Oguri, Michiyo; Chimetto, Luciane; Campeão, Mariana E; Thompson, Fabiano L; Thompson, Cristiane C

2014-02-01

283

Susceptibility to antibiotics of Vibrio sp. AO1 growing in pure culture or in association with its hydroid host Aglaophenia octodonta (Cnidaria, Hydrozoa).  

PubMed

Vibrio harveyi is the major causal organism of vibriosis, causing potential devastation to diverse ranges of marine invertebrates over a wide geographical area. These microorganisms, however, are phenotypically diverse, and many of the isolates are also resistant to multiple antibiotics. In a previous study, we described a previously unknown association between Vibrio sp. AO1, a luminous bacterium related to the species V. harveyi, and the benthic hydrozoan Aglaophenia octodonta. In this study, we analyzed the susceptibility to antibiotics (ampicillin, streptomycin, tetracycline, or co-trimoxazole = mix of sulfamethoxazole and trimetoprim) of Vibrio sp. AO1 growing in pure culture or in association with its hydroid host by using microcosm experiments. The results of minimum inhibitory concentration (MIC) experiments demonstrated that Vibrio sp. AO1 was highly resistant to ampicillin and streptomycin in pure culture. Nevertheless, these antibiotics, when used at sub-MIC values, significantly reduced the hydroid fluorescence. Co-trimoxazole showed the highest inhibitory effect on fluorescence of A. octodonta. However, in all treatments, the fluorescence was reduced after 48 h, but never disappeared completely around the folds along the hydrocaulus and at the base of the hydrothecae of A. octodonta when the antibiotic was used at concentration completely inhibiting growth in vitro. The apparent discrepancy between the MIC data and the fluorescence patterns may be due to either heterogeneity of the bacterial population in terms of antibiotic susceptibility or specific chemical-physical conditions of the hydroid microenvironment that may decrease the antibiotic susceptibility of the whole population. The latter hypothesis is supported by scanning electron microscope evidence for development of bacterial biofilm on the hydroid surface. On the basis of the results obtained, we infer that A. octodonta might behave as a reservoir of antibiotic multiresistant bacteria, increasing the risk of their transfer into aquaculture farms. PMID:19888625

Stabili, Loredana; Gravili, Cinzia; Boero, Ferdinando; Tredici, Salvatore M; Alifano, Pietro

2010-04-01

284

Integrative genome-scale metabolic analysis of Vibrio vulnificus for drug targeting and discovery  

Microsoft Academic Search

Although the genomes of many microbial pathogens have been studied to help identify effective drug targets and novel drugs, such efforts have not yet reached full fruition. In this study, we report a systems biological approach that efficiently utilizes genomic information for drug targeting and discovery, and apply this approach to the opportunistic pathogen Vibrio vulnificus CMCP6. First, we partially

Hyun Uk Kim; Soo Young Kim; Haeyoung Jeong; Tae Yong Kim; Jae Jong Kim; Hyon E Choy; Kyu Yang Yi; Joon Haeng Rhee; Sang Yup Lee

2011-01-01

285

[Antibiotic resistance and plasmid profiles of Vibrio isolates from cultured Sparus sarba].  

PubMed

A total of 51 potential pathogenic vibrios were isolated from moribund silver seabream Sparus sarba, which were collected from local fish farms of Hong Kong. All the isolates were classified and identified as 7 species by the API 20 E system and the scheme of Alsina & Blanch. These species were Vibrio alginolyticus (24 strains), Vibrio vulnificus (12 strains), Vibrio parahaemolyticus(7 strains), Vibrio logei(4 strains), Vibrio pelagius II(2 strains), Vibrio fluvialis (1 strains) and Vibrio meditterranei (1 strains). Among these isolates, the three predominant species (V. alginolyticus, V. vulnificus and V. parahaemolyticus) were confirmed to be virulent to sea bream by experimental challenge. All isolates were also screened for plasmid DNA by agarose gel electrophoresis and tested for susceptibility to 16 antimicrobial agents by the agar dilution method. Of the 51 isolates examined, all strains were sensitive to ceftriaxone, streptomycin, nalidixic acid and rifampicin, and almost all were sensitive to ceftazidime, netilimicin, chloramphenicol and sulfamethoxazole except one or two strains. Most isolates were resistant to ampicillin (60. 8%), cefuroxime (66.7%), amikacin(55%), kanamycin(58.8%) and trimethoprinm (76.5%). Fifteen of the 51 isolates harboured 1-4 plasmids, with sizes ranging from 9 to 123 kb. Both the plasmids and the associated antimicrobial resistance (ampicillin, cefuroxime and trimethoprim) of 9 isolates could be transferred to recipient by single-step conjugation, however, the frequencies were very low, ranging from 10(-11) to 10(-9). The present results indicate that resistance to these antibiotics is chromosomal. PMID:12555529

Li, J; Yie, J; Fu, W; Foo, R W; Hu, Y; Woo, N Y; Xu, H

1999-10-01

286

A survey of oysters (Crassostrea gigas) in New Zealand for Vibrio parahaemolyticus and Vibrio vulnificus.  

PubMed

A microbiological survey was conducted to determine the levels of total and pathogenic Vibrio parahaemolyticus (Vp) and Vibrio vulnificus (Vv) in Pacific oysters (Crassostrea gigas) collected from commercial growing areas in the North Island, New Zealand. The survey was intended to be geographically representative of commercial growing areas of Pacific oysters in New Zealand, while selecting the time frame most likely to coincide with the increased abundance of pathogenic vibrio species. Vp was detected in 94.8% of oyster samples examined (n=58) with a geometric mean concentration of 99.3 MPN/g, while Vv was detected in 17.2% of oyster samples examined with a geometric mean concentration of 7.4 MPN/g. The frequency of Vp positive samples was 1.7 fold greater than reported in a study conducted three decades ago in New Zealand. Potentially virulent (tdh positive) Vp was detected in two samples (3.4%, n=58) while no trh (another virulence marker) positive samples were detected. 16S rRNA genotype could be assigned only to 58.8% of Vv isolates (8:1:1 A:B:AB ratio, n=10). There was a good agreement [98.2% of Vp (n=280) and 94.4% of Vv (n=18) isolates] between molecular tests and cultivation based techniques used to identify Vibrio isolates and there was a significant (R(2)=0.95, P<0.001, n=18) linear relationship between the MPN estimates by real-time PCR and cultivation. There was no significant correlation between any of the environmental parameters tested and Vp or Vv concentrations. PMID:21501884

Kirs, M; Depaola, A; Fyfe, R; Jones, J L; Krantz, J; Van Laanen, A; Cotton, D; Castle, M

2011-05-27

287

Genome sequence of Vibrio splendidus : an abundant planctonic marine species with a large genotypic diversity  

Microsoft Academic Search

Vibrio splendidus is a dominant Vibrio species in seawater presenting a remarkable genetic diversity; several strains have been linked to invertebrate's mortality. We report the complete genome sequence of V. splendidus LGP32, an oyster pathogen, and its comparison with partial genome sequences from related strains. As is typical for the genus, V. splendidus LGP32 contains two chromosomes (3.29 and 1.67

Frédérique Le Roux; Mohamed Zouine; Nesrine Chakroun; Johan Binesse; Denis Saulnier; Christiane Bouchier; Nora Zidane; Laurence Ma; Christophe Rusniok; Aurélie Lajus; Carmen Buchrieser; Claudine Médigue; Martin F. Polz; Didier Mazel

2009-01-01

288

Identification of Vibrio harveyi Isolated from Diseased Asian Seabass Lates calcarifer by Use of 16S Ribosomal DNA Sequencing  

Microsoft Academic Search

The grow out of Asian seabass Lates calcarifer in marine net-cages is a popular aquaculture activity in Malaysia. Production of this species is greatly affected by the occurrence of vibriosis, which causes heavy mortality. Generally, young fish are more susceptible; they exhibit anorexia and skin darkening, followed by heavy mortality. The acutely affected older fish may also exhibit bloody lesions

Julian Ransangan; Saleem Mustafa

2009-01-01

289

Survival of Vibrio anguillarum and Vibrio salmonicida at different salinities.  

PubMed Central

The fish pathogenic bacteria Vibrio anguillarum and V. salmonicida showed the capacity to survive for more than 50 and 14 months, respectively, in seawater microcosms. A salinity of 5% proved lethal to V. anguillarum harvested in the late-exponential growth phase, whereas a salinity of 9% was lethal to the bacterium after it had been starved at a salinity of 30% for 67 days. The lethal salinity for V. salmonicida harvested in the late-exponential growth phase was probably in the vicinity of 10%. V. anguillarum and V. salmonicida were very sensitive to nalidixic acid. Direct determination of viable cells after incubation with nalidixic acid was not possible, since the cells did not elongate. Samples of V. salmonicida were double stained with fluorescein isothiocyanate-labeled antibodies and 4',6-diamidino-2-phenylindole. After 3 or 4 days of starvation, there was a discrepancy between the total numbers of cells as determined by immunofluorescence versus by staining with 4',6-diamidino-2-phenylindole. The immunofluorescence counts remained high, which indicated the presence of intact cell envelopes but leakage of DNA and other cytoplasm components. After 2 weeks of starvation, for some of the cells, the region stained with 4',6-diamidino-2-phenylindole (i.e., DNA) was markedly smaller than the cell envelope. I attributed this to a shrinkage of the cytoplasm or a confined nucleoid or both. V. anguillarum lost its exoproteolytic activity before 11 days of starvation.

Hoff, K A

1989-01-01

290

Research Note A Collagenase-Targeted Multiplex PCR Assay for Identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus  

Microsoft Academic Search

A multiplex PCR assay using three collagenase-targeted primer pairs for the species-specific detection of Vibrio algino- lyticus, Vibrio cholerae, and Vibrio parahaemolyticuswas developed. The results highlight the species specificity of the three primer sets designed. Because of the increasing importance of Vibrio spp. in human foodborne diseases, molecular approaches for routine microbial screening and monitoring of clinical, environmental, and food

ANGELA DI PINTO; GIUSEPPINA CICCARESE; GIUSEPPINA TANTILLO; DOMENICO CATALANO; VITO TONY FORTE

291

Role of the virulence plasmid pR99 and the metalloprotease Vvp in resistance of Vibrio vulnificus serovar E to eel innate immunity  

Microsoft Academic Search

Vibrio vulnificus biotype 2 serovar E (VSE) is a bacterial pathogen that produces a haemorrhagic septicaemia called vibriosis in eels. Its ability to grow in blood is conferred by a recently described virulence plasmid [Lee CT, Amaro C, Wu KM, Valiente E, Chang YF, Tsai SF, et al. A common virulence plasmid in biotype 2 Vibrio vulnificus and its dissemination

E. Valiente; C. T. Lee; J. Lamas; L. Hor; C. Amaro

2008-01-01

292

Vibrio cholerae O139 Conjugate Vaccines: Synthesis and Immunogenicity of V. cholerae O139 Capsular Polysaccharide Conjugates with Recombinant Diphtheria Toxin Mutant in Mice  

Microsoft Academic Search

Epidemiologic and experimental data provide evidence that a critical level of serum immunoglobulin G (IgG) antibodies to the surface polysaccharide of Vibrio cholerae O1 (lipopolysaccharide) and of Vibrio cholerae O139 (capsular polysaccharide (CPS)) is associated with immunity to the homologous pathogen. The immunoge- nicity of polysaccharides, especially in infants, may be enhanced by their covalent attachment to proteins (conjugates). Two

ZUZANA KOSSACZKA; JOSEPH SHILOACH; VIRGINIA JOHNSON; DAVID N. TAYLOR; RICHARD A. FINKELSTEIN; JOHN B. ROBBINS; SHOUSUN C. SZU

2000-01-01

293

Isolation of Vibrio vulnificus from Seawater and Emerging Vibrio vulnificus Septicemia on Jeju Island  

PubMed Central

Vibrio vulnificus is an opportunistic human pathogen, transmitted from seawater, raw oyster, and shellfish and responsible for severe septicemia. We studied V. vulnificus from surface seawater around Jeju Island between 2010 and 2011. In 2010, V. vulnificus was isolated and V. vulnificus septicemia was reported. Surface seawater temperature is an important factor for growth of V. vulnificus, and here we showed that high surface seawater temperature may influence growth of V. vulnificus and occurrence of emerging V. vulnificus septicemia on Jeju Island. This is the first report of isolation of V. vulnificus and emerging V. vulnificus septicemia on Jeju Island.

Lee, Keun Hwa; Kim, Young Ree; Pang, Ig-Chan

2014-01-01

294

Distribution of Vibrio vulnificus and Other Lactose-Fermenting Vibrios in the Marine Environment  

PubMed Central

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 H2S 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.

Oliver, James D.; Warner, Robert A.; Cleland, David R.

1983-01-01

295

Temporal Quorum-Sensing Induction Regulates Vibrio cholerae Biofilm Architecture  

Microsoft Academic Search

Vibrio cholerae, the pathogen that causes cholera, also survives in aqueous reservoirs, probably in the form of biofilms. Quorum sensing negatively regulates V. cholerae biofilm formation through HapR, whose expres- sion is induced at a high cell density. In this study, we show that the concentration of the quorum-sensing signal molecule CAI-1 is higher in biofilms than in planktonic cultures.

Zhi Liu; Fiona R. Stirling; Jun Zhu

2007-01-01

296

Identification of non-coding RNAs in environmental vibrios.  

PubMed

The discovery of non-coding RNA (ncRNA) has been mainly limited to laboratory model systems and human pathogenic bacteria. In this study, we begin to explore the ncRNA diversity in four recently sequenced environmental Vibrio species (Vibrio alginolyticus 40B, Vibrio communis 1DA3, Vibrio mimicus VM573 and Vibrio campbellii BAA-1116) by performing in silico searches using Infernal and Rfam for the identification of putative ncRNA-encoding genes. This search method resulted in the identification of 31-38 putative ncRNA genes per species and the total ncRNA catalogue spanned an assortment of regulatory mechanisms (riboswitches, cis-encoded ncRNAs, trans-encoded ncRNAs, modulators of protein activity, ribonucleoproteins, transcription termination ncRNAs and unknown). We chose to experimentally validate the identifications for V. campbellii BAA-1116 using a microarray-based expression profiling strategy. Transcript hybridization to tiled probes targeting annotated V. campbellii BAA-1116 intergenic regions revealed that 21 of the 38 predicted ncRNA genes were expressed in mid-exponential-phase cultures grown in nutrient-rich medium. The microarray findings were confirmed by testing a subset of three highly expressed (6S, tmRNA and TPP-2) and three moderately expressed (CsrB, GcvB and purine) ncRNAs via reverse transcription PCR. Our findings provide new information on the diversity of ncRNA in environmental vibrios while simultaneously promoting a more accurate annotation of genomic intergenic regions. PMID:20447992

Silveira, Ana Cristina G; Robertson, Kelly L; Lin, Baochuan; Wang, Zheng; Vora, Gary J; Vasconcelos, Ana Tereza R; Thompson, Fabiano L

2010-08-01

297

Does Vibrio vulnificus present a health threat to Canadians?  

PubMed Central

OBJECTIVE: To review recent data on Vibrio vulnificus and its properties, characteristics of disease and epidemiology, sources of infection, population at risk, infectious dose, documented cases of infection and health risk from V vulnificus infection in Canada. DATA SOURCE: A MEDLINE and CURRENT CONTENTS search (1981 through September 1996) using the main heading ‘Vibrio vulnificus’, ‘Vibrio species’, ‘seafood’, etc. Relevant articles were also selected from the literature collection in the authors’ laboratory. STUDY SELECTION AND DATA EXTRACTION: The authors judged articles relevant to the objective of the paper and selected them for a review. DATA SYNTHESIS: V vulnificus, an important cause of septicemia, wound infections and gastroenteritis, is considered to be one of the most invasive and rapidly lethal human pathogens. Molluscan shellfish concentrate this organism from warm seawater and present the greatest danger to consumers. Infections with this pathogen have been reported throughout the world. Most deaths have resulted from the consumption of raw or undercooked oysters, fewer from contact with seawater. Individuals with underlying disease, particularly those with liver diseases and iron overload, are the most susceptible. CONCLUSION: The two reported cases of V vulnificus wound infection in Canada might not represent the real situation. Infection with this organism may go unrecognized, unreported or simply may not occur. Medical professionals need to become aware of this pathogen and the dire consequences of infection in individuals with underlying disease.

Stavric, Stanislava; Buchanan, Barbara

1997-01-01

298

Long-term effects of ocean warming on vibrios  

NASA Astrophysics Data System (ADS)

Vibrios are a major source of human disease, play an important role in the ecology and health of marine animals and are regarded as an abundant fraction of culturable bacteria of the ocean. There has been a considerable global effort to reduce the risk of Vibrio infections and yet in most countries both human and non-human illnesses associated with these bacteria are increasing. The cause of this increase is not known, but since vibrios are strongly thermodependant there is good reason to believe that global warming may have contributed. To investigate this possibility we examined historical samples from the Continuous Plankton Recorder (CPR) archive using advanced molecular analysis and pyrosequencing. For the first time we were able to recover environmental DNA from CPR samples that had been stored for up to ~50 years in a formalin-fixed format, which is suitable for molecular analyses of the associated prokaryotic community. To overcome the problem of DNA degradation due to the sample age and storage in formalin we develop an unbiased index of abundance for Vibrio quantification in CPR samples termed a 'relative Vibrio Abundance Index' (VAI). VAI is defined as the ratio of Vibrio spp. cells to total bacterial cells assessed by Real-Time PCR using genus-specific and universal primers, respectively, producing small amplicons of similar size (~100bp). We assessed VAI index on 55 samples (each representing 10 nautical miles tow equal to 3 m3 of filtered sewater) collected in August by the CPR survey in the North Sea from off the Rhine and Humber estuaries between 1961 to 2005 showing that the genus Vibrio has increased in prevalence in the last 44 years and that this increase is correlated significantly, during the same period, with warming sea surface temperature. In addition, by applying deep sequencing analysis of a subset of these samples we provide evidence that bacteria belonging to the genus Vibrio, including the human pathogen V. cholerae, not only increased in occurrence over the last half century in the southern North Sea, but also prevailed within the particle associated bacterial community of coastal marine waters. These findings provide support for the view that global warming may have a strong impact on the composition of marine bacterial communities with important implications for human and animal health into the future.

Pruzzo, C.; Pezzati, E.; Brettar, I.; Reid, P. C.; Colwell, R.; Höfle, M. G.; vezzulli, L.

2012-12-01

299

HlyU acts as an H-NS antirepressor in the regulation of the RTX toxin gene essential for the virulence of the human pathogen Vibrio vulnificus CMCP6  

PubMed Central

Summary In Vibrio vulnificus, HlyU up-regulates the expression of the large RTX toxin gene. In this work we identified the binding site of HlyU to -417 to -376 bp of the rtxA1 operon transcription start site. lacZ fusions for a series of progressive deletions from the rtxA1 operon promoter showed that transcriptional activity increased independently of HlyU when its binding site was absent. Thus HlyU must regulate the rtxA1 operon expression by antagonizing a negative regulator. Concomitantly we found that an hns mutant resulted in an increase in the expression of the rtxA1 operon genes. Multiple copies of HlyU can increase the promoter activity only in the presence of H-NS underscoring the hypothesis that HlyU must alleviate the repression by this protein. H-NS binds to a region that extends upstream and downstream of the rtxA1 operon promoter. In the upstream region it binds to five AT-rich sites of which two overlap the HlyU binding site. Competitive footprinting and gel shift data demonstrate HlyU’s higher affinity as compared to H-NS resulting in the de-repression and a corresponding increased expression of the rtxA1 operon.

Liu, Moqing; Naka, Hiroaki; Crosa, Jorge H.

2009-01-01

300

Distribution of the Luminous Bacterium Beneckea harveyi in a Semitropical Estuarine Environment.  

PubMed

Bioluminescent bacteria were found in the water column, sediment, shrimp, and gastrointestinal tract of marine fishes from the semitropical estuarine environment of the East Lagoon, Galveston Island, Tex. Populations in the water column decreased during cold weather while sedimentary populations persisted. The highest percentages of luminous organisms were isolated from the gastrointestinal tract of marine fishes, where they persisted during 5 days of starvation. The presence of chitin temporarily increased intestinal populations. All isolates were Beneckea harveyi, whose natural habitat appears to be the gut of fishes and whose free-living reservoir appears to be marine sediments. PMID:16345465

O'brien, C H; Sizemore, R K

1979-11-01

301

Distribution of the Luminous Bacterium Beneckea harveyi in a Semitropical Estuarine Environment  

PubMed Central

Bioluminescent bacteria were found in the water column, sediment, shrimp, and gastrointestinal tract of marine fishes from the semitropical estuarine environment of the East Lagoon, Galveston Island, Tex. Populations in the water column decreased during cold weather while sedimentary populations persisted. The highest percentages of luminous organisms were isolated from the gastrointestinal tract of marine fishes, where they persisted during 5 days of starvation. The presence of chitin temporarily increased intestinal populations. All isolates were Beneckea harveyi, whose natural habitat appears to be the gut of fishes and whose free-living reservoir appears to be marine sediments.

O'Brien, Catherine H.; Sizemore, Ronald K.

1979-01-01

302

Vibrio Fischeri Symbiosis Gene Regulation.  

National Technical Information Service (NTIS)

The goals of this project were to investigate the molecular mechanisms controlling luminescence gene expression of the symbiotic bioluminescent bacterium Vibrio fischeri; and to identify and investigate the regulation of other symbiosis functions in this ...

P. V. Dunlap

1989-01-01

303

Abundance and distribution of Vibrio cholerae, V. parahaemolyticus, and V. vulnificus following a major freshwater intrusion into the Mississippi Sound.  

PubMed

In response to a major influx of freshwater to the Mississippi Sound following the opening of the Bonnet Carre Spillway, water samples were collected from three sites along the Mississippi shoreline to assess the impact of altered salinity on three pathogenic Vibrio species. Salinity readings across the affected area during the 2011 sample period ranged from 1.4 to 12.9 ppt (mean = 7.0) and for the 2012 sample period from 14.1 to 23.6 ppt (mean = 19.8). Analyses of the data collected in 2011 showed a reduction in densities of Vibrio parahaemolyticus and Vibrio vulnificus with a concurrent increase of Vibrio cholerae numbers, with V. cholerae becoming the only Vibrio detected once salinity readings dropped to 6 ppt. Follow-up samples taken in 2012 after recovery of the salinity in the sound showed that the relative densities of the three pathogenic vibrios had reverted back to normal levels. This study shows that although the spillway was open but a few weeks and the effects were therefore time limited, the Mississippi River water had a profound, if temporary, effect on Vibrio ecology in the Mississippi Sound. PMID:23494573

Griffitt, Kimberly J; Grimes, D Jay

2013-04-01

304

Reverse Transcription-Multiplex PCR Assay for Simultaneous Detection of Escherichia coli O157: H7, Vibrio cholerae O1, and Salmonella Typhi  

Microsoft Academic Search

Background: Escherichia coli O157:H7, Vibrio cholerae O1, and Salmonella Typhi are pathogenic bacteria that can be found in contaminated water supplies through- out the world. No currently available assays can simul- taneously detect and identify all three pathogens. Our aim was to develop a rapid and reliable technique for simultaneous detection of these pathogens. Methods: Four unique genes were chosen

Nicole J. Morin; Zhilong Gong; Xing-Fang Li

2004-01-01

305

Detection of toxins produced by vibrio fluvialis.  

PubMed Central

The results of studies with cell-free extracts and culture supernatant fluids of Vibrio fluvialis (a recently recognized, potential enteric pathogen for humans) grown in the absence and presence of lincomycin indicated that the bacterium could produce (i) a factor which causes CHO cell elongation (CEF) similar to that elicited by V. cholerae enterotoxin and by the heat-labile enterotoxin of Escherichia coli, (ii) cytolysin(s) active against erythrocytes, (iii) nonhemolytic, CHO cell-killing factor(s), and (iv) protease(s) active against azocasein. The CEF was heat labile and ammonium sulfate precipitable, and it had an isoelectric point (estimated by sucrose density gradient electrofocusing) and molecular weight (estimated by gel filtration) of about 5.1 and 135,000, respectively. Images

Lockwood, D E; Kreger, A S; Richardson, S H

1982-01-01

306

Polyphasic Taxonomy of the Genus Vibrio: Numerical Taxonomy of Vibrio cholerae, Vibrio parahaemolyticus, and Related Vibrio Species  

PubMed Central

A set of 86 bacterial cultures, including 30 strains of Vibrio cholerae, 35 strains of V. parahaemolyticus, and 21 representative strains of Pseudomonas, Spirillum, Achromobacter, Arthrobacter, and marine Vibrio species were tested for a total of 200 characteristics. Morphological, physiological, and biochemical characteristics were included in the analysis. Overall deoxyribonucleic acid (DNA) base compositions and ultrastructure, under the electron microscope, were also examined. The taxonomic data were analyzed by computer by using numerical taxonomy programs designed to sort and cluster strains related phenetically. The V. cholerae strains formed an homogeneous cluster, sharing overall S values of ?75%. Two strains, V. cholerae NCTC 30 and NCTC 8042, did not fall into the V. cholerae species group when tested by the hypothetical median organism calculation. No separation of “classic” V. cholerae, El Tor vibrios, and nonagglutinable vibrios was observed. These all fell into a single, relatively homogeneous, V. cholerae species cluster. V. parahaemolyticus strains, excepting 5144, 5146, and 5162, designated members of the species V. alginolyticus, clustered at S ?80%. Characteristics uniformly present in all the Vibrio species examined are given, as are also characteristics and frequency of occurrence for V. cholerae and V. parahaemolyticus. The clusters formed in the numerical taxonomy analyses revealed similar overall DNA base compositions, with the range for the Vibrio species of 40 to 48% guanine plus cytosine. Generic level of relationship of V. cholerae and V. parahaemolyticus is considered dubious. Intra- and intergroup relationships obtained from the numerical taxonomy studies showed highly significant correlation with DNA/DNA reassociation data. Images

Colwell, R. R.

1970-01-01

307

Vaccination with three inactivated pathogens of cobia ( Rachycentron canadum) stimulates protective immunity  

Microsoft Academic Search

Cobia (Rachycentron canadum), a warm water fish recently commercially cultured in Taiwan, has encountered severe mortalities with survival rates often below 20%. The major causative pathogens were Vibrio alginolyticus, Vibrio parahaemolyticus and Photobacterium damselae subsp. piscicida. We prepared a combined three inactivated bacterins antigen preparation and vaccinated cobia. This mixture of bacterins was safe, and the immune response in cobia

John Han-You Lin; Tzong-Yueh Chen; Ming-Shyan Chen; Huey-En Chen; Ruey-Liang Chou; Tzyy-Ing Chen; Mao-Sen Su; Huey-Lang Yang

2006-01-01

308

Targeting the Replication Initiator of the Second Vibrio Chromosome: Towards Generation of Vibrionaceae-Specific Antimicrobial Agents  

PubMed Central

The Vibrionaceae is comprised of numerous aquatic species and includes several human pathogens, such as Vibrio cholerae, the cause of cholera. All organisms in this family have two chromosomes, and replication of the smaller one depends on rctB, a gene that is restricted to the Vibrionaceae. Given the increasing prevalence of multi-drug resistance in pathogenic vibrios, there is a need for new targets and drugs to combat these pathogens. Here, we carried out a high throughput cell-based screen to find small molecule inhibitors of RctB. We identified a compound that blocked growth of an E. coli strain bearing an rctB-dependent plasmid but did not influence growth of E. coli lacking this plasmid. This compound, designated vibrepin, had potent cidal activity against V. cholerae and inhibited the growth of all vibrio species tested. Vibrepin blocked RctB oriCII unwinding, apparently by promoting formation of large non-functional RctB complexes. Although vibrepin also appears to have targets other than RctB, our findings suggest that RctB is an attractive target for generation of novel antibiotics that only block growth of vibrios. Vibrio-specific agents, unlike antibiotics currently used in clinical practice, will not engender resistance in the normal human flora or in non-vibrio environmental microorganisms.

Yamaichi, Yoshiharu; Duigou, Stephane; Shakhnovich, Elizabeth A.; Waldor, Matthew K.

2009-01-01

309

Diversity and dynamics of the Vibrio community in well water used for drinking in Guinea-Bissau (West Africa).  

PubMed

Bacteria of the genus Vibrio are ubiquitous in aquatic environments and can be found either in culturable or in a viable but nonculturable (VBNC) state. The genus comprises many pathogenic species accountable for water and food-borne diseases that prove to be fatal, especially in developing countries, as in Guinea-Bissau (West Africa), where cholera is endemic. In order to ascertain the abundance and structure of Vibrio spp. community in well waters that serve as the sole source of water for the population, quantitative polymerase chain reaction (qPCR), PCR-denaturant gradient gel electrophoresis (DGGE), and cloning approaches were used. Results suggest that Vibrio spp. were present throughout the year in acidic, freshwater wells with a seasonal community composition shift. Vibrio spp. abundance was in accordance with the abundance found in coastal environments. Sequences closely related to pathogenic Vibrio species were retrieved from well water revealing exposure of the population to such pathogens. pH, ammonium, and turbidity, regulated by the rain pattern, seem to be the variables that contributed mostly to the shaping and selection of the Vibrio spp. community. These results reinforce the evidence for water monitoring with culture-independent methods and the clear need to create/recover water infrastructures and a proper water resources management in West African countries with similar environmental conditions. PMID:24859857

Machado, A; Bordalo, A A

2014-09-01

310

Detection and quantification of Vibrio populations using denaturant gradient gel electrophoresis.  

PubMed

Bacteria affiliated with the genus Vibrio are endemic in marine and estuarine ecosystems and are also found in many freshwater environments. Vibrios can enter viable but non-culturable states and since many species are pathogenic, there is a great need for culture-independent methods that identify and quantify multiple Vibrio populations. We adopted Vibrio-specific 16S rRNA-directed primers and a competitive PCR protocol (QC-PCR; [Thompson, J.R., Randa, M.A., Marcelino, L.A., Tomita-Mitchell, A., Lim, E., Polz, M.F., 2004b. Diversity and dynamics of a North Atlantic coastal Vibrio community. Appl. Environ. Microbiol. 70, 4103-4110]) for separation and quantification of Vibrio populations using denaturant gradient gel electrophoresis (DGGE). Sixteen Vibrio isolates and eight environmental samples were used to assess the precision and resolution of the method. A 45-70% gradient of Urea and formamide enabled separation of Vibrio populations with single nucleotide differences in the amplified fragment. A titration curve for the QC-PCR-DGGE, verified by amending surface water bacterioplankton samples with up to 3 x 10(5)Vibrio cholerae cells, could be approximated by a linear regression of log-transformed values (R(2)=0.96). The limit of detection for single populations was 180 cells per extracted sample or about 4 cells per PCR reaction. Environmental samples from the southern Stockholm archipelago in the Baltic Sea and the more saline coastal waters of Skagerrak each carried between 2 and 6 Vibrio populations, and there were major differences between the locations. Notably, multiple Vibrio populations could be detected and quantified against a background of native bacterioplankton exceeding Vibrio population abundance by more than 6 orders of magnitude. Putative identification based on migration in the DGGE gel was verified by parallel cloning and sequencing of PCR products, and representative clones were also characterized by DGGE. This general approach could also be useful for targeting other phylogenetically constrained bacterial groups and assess their abundance and distribution in complex environmental settings. PMID:16730823

Eiler, Alexander; Bertilsson, Stefan

2006-11-01

311

Draft Genome Sequence of Israeli Outbreak-Associated Vibrio vulnificus Biotype 3 Clinical Isolate BAA87.  

PubMed

Vibrio vulnificus is a seafood-associated pathogen that causes severe wound and intestinal infections. Biotype 3 of V. vulnificus emerged in 1996 as the cause of an Israeli outbreak associated with the handling of infected tilapia. Here, we describe the whole-genome sequence of the ATCC biotype 3 clinical isolate BAA87 (CDC9530-96). PMID:24652973

Phillips, Kelsey E; Schipma, Matthew J; Satchell, Karla J F

2014-01-01

312

Distribution of Virulence Genes in Clinical and Environmental Vibrio cholerae Strains in Bangladesh  

PubMed Central

Vibrio cholerae, an environmental organism, is a facultative human pathogen. Here, we report the virulence profiles, comprising 18 genetic markers, of 102 clinical and 692 environmental V. cholerae strains isolated in Bangladesh between March 2004 and January 2006, showing the variability of virulence determinants within the context of public health.

Hasan, Nur A.; Ceccarelli, Daniela; Grim, Christopher J.; Taviani, Elisa; Choi, Jinna; Sadique, Abdus; Alam, Munirul; Siddique, Abul K.; Sack, R. Bradley; Huq, Anwar

2013-01-01

313

Vibrio natriegens: A Rapidly Growing Micro-Organism Ideally Suited for Class Experiments  

ERIC Educational Resources Information Center

Describes five microbiological experiments using the marine organism Vibrio natriegens. This organism is highly suitable for laboratory work because it is non-pathogenic and grows extremely rapidly, having the distinction of the lowest mean generation time yet recorded (9.8 minutes). (JR)

Mullenger, L.; Gill, Nijole R.

1973-01-01

314

Characterization of Vibrio fluvialis-Like Strains Implicated in Limp Lobster Disease  

Microsoft Academic Search

Studies were undertaken to characterize and determine the pathogenic mechanisms involved in a newly described systemic disease in Homarus americanus (American lobster) caused by a Vibrio fluvialis-like micro- organism. Nineteen isolates were obtained from eight of nine lobsters sampled. Biochemically, the isolates resembled V. fluvialis, and the isolates grew optimally at 20°C; none could grow at temperatures above 23°C. The

B. D. Tall; S. Fall; M. R. Pereira; M. Ramos-Valle; S. K. Curtis; M. H. Kothary; D. M. T. Chu; S. R. Monday; L. Kornegay; T. Donkar; D. Prince; R. L. Thunberg; K. A. Shangraw; D. E. Hanes; F. M. Khambaty; K. A. Lampel; J. W. Bier; R. C. Bayer

2003-01-01

315

Antagonistic Interactions among Marine Bacteria Impede the Proliferation of Vibrio cholerae  

Microsoft Academic Search

Changes in global climate have raised concerns about the emergence and resurgence of infectious diseases. Vibrio cholerae is a reemerging pathogen that proliferates and is transported on marine particles. Patterns of cholera outbreaks correlate with sea surface temperature increases, but the underlying mechanisms for rapid proliferation of V. cholerae during ocean warming events have yet to be fully elucidated. In

Richard A. Long; David C. Rowley; Eric Zamora; Jiayuan Liu; Douglas H. Bartlett; Farooq Azam

2005-01-01

316

USER Friendly Cloning Coupled with Chitin-Based Natural Transformation Enables Rapid Mutagenesis of Vibrio vulnificus  

Microsoft Academic Search

Vibrio vulnificus is a bacterial contaminant of shellfish and causes highly lethal sepsis and destructive wound infections. A definitive identification of virulence factors using the molecular version of Koch's postulates has been hindered because of difficulties in performing molecular genetic analysis of this opportunistic pathogen. For example, conjugation is required to introduce plasmid DNA, and allelic exchange suicide vectors that

Paul A. Gulig; Matthew S. Tucker; Patrick C. Thiaville; Jennifer L. Joseph; Roslyn N. Brown

2009-01-01

317

Response of Vibrio parahaemolyticus 03:K6 to a hot water\\/cold shock pasteurization process  

Microsoft Academic Search

Vibrio vulnificus and V. parahaemolyticus are natural inhabitants of estuarine environments world wide. Pathogenic strains of these bacteria are often transmitted to humans through consumption of raw oysters, which flourish in the same estuaries. Previous studies reported the effective use of hot water pasteurization followed by cold shock to eliminate from raw oysters naturally and artificially incurred environmental strains of

L. S. Andrews; S. DeBlanc; C. D. Veal; D. L. Park

2003-01-01

318

Vibrio furnissii: an Unusual Cause of Bacteremia and Skin Lesions after Ingestion of Seafood?†  

PubMed Central

Vibrio furnissii in the blood is rarely reported, which may explain why clinical features of bloodstream infections with this organism have not been described. We describe a patient who developed skin lesions and V. furnissii bacteremia and was successfully treated with fluoroquinolones. V. furnissii may be a serious pathogen in patients with underlying comorbidities who are exposed to seafood.

Derber, Catherine; Coudron, Philip; Tarr, Cheryl; Gladney, Lori; Turnsek, Maryann; Shankaran, Shivanjali; Wong, Edward

2011-01-01

319

Incidence of highly genetically diversified Vibrio parahaemolyticus in seafood imported from Asian countries  

Microsoft Academic Search

Vibrio parahaemolyticus is an important pathogen of humans and aquacultured animals, especially in Asian countries. In this study, we examined 686 samples of seafood imported from Hong Kong, Indonesia, Thailand and Vietnam for V. parahaemolyticus. V. parahaemolyticus was recovered from 315 (45.9%) samples. The incidence of V. parahaemolyticus in products from Hong Kong and Thailand was markedly higher than the

H.-C Wong; M.-C Chen; S.-H Liu; D.-P Liu

1999-01-01

320

Comparing the efficiency of chitosan with chlorine for reducing Vibrio parahaemolyticus in shrimp  

Microsoft Academic Search

Thailand is one of the leading exporters of frozen shrimp to many countries. Chlorine is the decontaminating agent most frequently used in the frozen shrimp industries to kill potential pathogens. However, long time contact to chlorine causes severe respiratory tract damage. In this study, chitosan was compared to chlorine for reducing Vibrio parahaemolyticus. In vitro investigation, chitosan could reduce more

S. Chaiyakosa; W. Charernjiratragul; K. Umsakul; V. Vuddhakul

2007-01-01

321

Vibrio Zinc-Metalloprotease Causes Photoinactivation of Coral Endosymbionts and Coral Tissue Lesions  

PubMed Central

Background Coral diseases are emerging as a serious threat to coral reefs worldwide. Of nine coral infectious diseases, whose pathogens have been characterized, six are caused by agents from the family Vibrionacae, raising questions as to their origin and role in coral disease aetiology. Methodology/Principal Findings Here we report on a Vibrio zinc-metalloprotease causing rapid photoinactivation of susceptible Symbiodinium endosymbionts followed by lesions in coral tissue. Symbiodinium photosystem II inactivation was diagnosed by an imaging pulse amplitude modulation fluorometer in two bioassays, performed by exposing Symbiodinium cells and coral juveniles to non-inhibited and EDTA-inhibited supernatants derived from coral white syndrome pathogens. Conclusion/Significance These findings demonstrate a common virulence factor from four phylogenetically related coral pathogens, suggesting that zinc-metalloproteases may play an important role in Vibrio pathogenicity in scleractinian corals.

Sussman, Meir; Mieog, Jos C.; Doyle, Jason; Victor, Steven; Willis, Bette L.; Bourne, David G.

2009-01-01

322

Identification of Outer Membrane Proteins Altered in Response to UVC-Radiation in Vibrio parahaemolyticus and Vibrio alginolyticus.  

PubMed

Vibrio parahaemolyticus and V. alginolyticus, marine foodborne pathogens, were treated with UVC-radiation (240 J/m(2)) to evaluate alterations in their outer membrane protein profiles. Outer membrane protein patterns of UVC-irradiated bacteria were found altered when analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Altered proteins were identified by mass spectrometry (MS and MS/MS) and analysis revealed that OmpW, OmpA, Long-chain fatty acid transport protein, Outer membrane receptor protein, Putative uncharacterized protein VP0167, Maltoporin (lamB), Polar flagellin B/D, Agglutination protein Peptidoglycan-associated lipoprotein and MltA-interacting protein MipA were appeared, thereby they can be considered as UVC-stress proteins in some vibrios. In addition, expression of OmpK decreased to non-detectable level. Furthermore, we observed a decrease or an increase in the expression level of other outer membrane proteins. PMID:24293727

Abdallah, Fethi Ben; Lagha, Rihab; Ellafi, Ali; Namane, Abdelkader; Rousselle, Jean-Claude; Lenormand, Pascal; Kallel, Héla

2012-12-01

323

A collagenase-targeted multiplex PCR assay for identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus.  

PubMed

A multiplex PCR assay using three collagenase-targeted primer pairs for the species-specific detection of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus was developed. The results highlight the species specificity of the three primer sets designed. Because of the increasing importance of Vibrio spp. in human foodborne diseases, molecular approaches for routine microbial screening and monitoring of clinical, environmental, and food samples also have become more important. The results of this study indicate that the gene coding for collagenase should be used as an alternative molecular target to discriminate among the three Vibrio species. PMID:15690817

Di Pinto, Angela; Ciccarese, Giuseppina; Tantillo, Giuseppina; Catalano, Domenico; Forte, Vito Tony

2005-01-01

324

A Role for the Mannose-Sensitive Hemagglutinin in Biofilm Formation by Vibrio cholerae El Tor  

Microsoft Academic Search

While much has been learned regarding the genetic basis of host-pathogen interactions, less is known about the molecular basis of a pathogen's survival in the environment. Biofilm formation on abiotic surfaces represents a survival strategy utilized by many microbes. Here it is shown that Vibrio cholerae El Tor does not use the virulence-associated toxin-coregulated pilus to form biofilms on borosilicate

PAULA I. WATNICK; KARLA JEAN FULLNER; ROBERTO KOLTER

1999-01-01

325

Vibrio cholerae Interactions with the Gastrointestinal Tract: Lessons from Animal Studies  

Microsoft Academic Search

\\u000a \\u000a Vibrio cholerae is a curved Gram-negative rod that causes the diarrheal disease cholera. One hundred and twenty five years of study of V. cholerae microbiology have made this lethal pathogen arguably the most well-understood non-invasive mucosal pathogen. Over the past\\u000a 25 years, modern molecular techniques have permitted the identification of many genes and cellular processes that are critical\\u000a for V.

Jennifer M. Ritchie; Matthew K. Waldor

326

Characterization of a New Plasmid-Like Prophage in a Pandemic Vibrio parahaemolyticus O3:K6 Strain  

Microsoft Academic Search

Vibrio parahaemolyticus is a common food-borne pathogen that is normally associated with seafood. In 1996, a pandemic O3:K6 strain abruptly appeared and caused the first pandemic of this pathogen to spread throughout many Asian countries, America, Europe, and Africa. The role of temperate bacteriophages in the evolution of this pathogen is of great interest. In this work, a new temperate

Shih-Feng Lan; Chung-Ho Huang; Chuan-Hsiung Chang; Wei-Chao Liao; I-Hsuan Lin; Wan-Neng Jian; Yueh-Gin Wu; Shau-Yan Chen; Hin-chung Wong

2009-01-01

327

Detection, isolation, and identification of Vibrio cholerae from the environment.  

PubMed

Microbiological techniques for sampling the aquatic realm have become increasingly sophisticated, especially with advances in molecular biology. These techniques have been used to detect microorganisms that cannot be cultured by conventional bacteriological methods. This has resulted in a deeper and a clearer understanding of the ecology and epidemiology of microorganisms. Important advances have been made in isolation, detection, and identification of Vibrio cholerae over the past decade. The understanding that V. cholerae, like several other pathogenic bacteria, can enter into a state known as "viable but nonculturable" (VBNC) provided important clues on the epidemiology of the pathogen and its ability to cause sudden explosive epidemics at multiple places almost simultaneously. The advances in techniques have also allowed investigators to discern the intricate aspects of the ecology of this pathogen in the aquatic world. In this unit, we present the most accepted methods for the isolation and detection of V. cholerae. PMID:18770592

Huq, Anwar; Grim, Christopher; Colwell, Rita R; Nair, G Balakrish

2006-09-01

328

Detection of Vibrio splendidus and related species in Chamelea gallina sampled in the Adriatic along the Abruzzi coastline.  

PubMed

Vibrio species are an important and widespread component of marine microbial communities. Some Vibrio strains are potentially pathogenic to marine vertebrates and invertebrates. The aim of this study was to identify vibrios, in particular Vibrio splendidus and related species, isolated from clams (Chamelea gallina) collected along the coasts of the Abruzzi region from May to October 2007. The isolates obtained were phenotyped and classified as belonging to the genus Vibrio. The strains underwent biochemical testing in accordance with Alsina's scheme for V. splendidus identification. Molecular analysis of the 16S-23S intergenic space region and recA gene was used to identify V. splendidus and related species. All the samples examined were found to contain halophylic Vibrio species, with V. alginolyticus, V. splendidus-related species and V. mediterranei most commonly found. A polymerase chain reaction of the 16S-23S intergenic space region and sequencing of the recA gene from isolates confirmed that phenotyping of Vibrio species is not sufficient to distinguish between different species. Differentiation of the highly related species among V. splendidus-related clusters remains an important issue. In this regard, our data suggests sequencing the recA genes was far more discriminatory than sequencing 16S rDNA for this purpose. PMID:21947974

Torresi, Marina; Acciari, Vicdalia A; Piano, Annamaria; Serratore, Patrizia; Prencipe, Vincenza; Migliorati, Giacomo

2011-01-01

329

Intracellular survival of Vibrio anguillarum  

Microsoft Academic Search

The intracellular survival of a Vibrio anguillarum strain, ingested by head kidney phagocytes and peripheral leukocytes of grouper, Epinephelus awoara, was assessed in vitro by comparing with that of V. parahaemolyticus and Staphylococcus albus. There was an increase in numbers of V. alginolyticus in macrophages from head kidney and peripheral leukocytes during the first 30min after infection, followed by a

Wenbo Chen; Yingxue Qin; Qingpi Yan

2010-01-01

330

VIBRIO VULNIFICUS EDUCATION WORKSHOPS / MATERIALS  

EPA Science Inventory

This project will promote Vibrio vulnificus education on-line continuing medical education units to health care professionals that counsel and care for at-risk individuals. The Florida Department of Agriculture and Consumer Services will purchase advertisement and promotion in me...

331

CT gene modulate differential expression of chitinase gene under variant habitats in Vibrio cholerae  

PubMed Central

Objective To investigate the interrelation of cholera toxin gene (CT gene) in expression of chitinase gene under different pH conditions among pathogenic and Non-pathogenic strains of Vibrio cholera (V. cholera). Methods The chitinase assay well diffusion method and calorimetric chitinase assay were performed. Further, time depended chitinase activity among pathogenic and nonpathogenic strain was evaluated with control as Escherichia coli. The expressed protein in variant environment was purified by cascade of chromatographic techniques. The partially purified protein was analyzed by SDS-PAGE in both the strain of V. cholera. Results The results have shown differential expression of chitinase gene among vibrio in time depended chitinase activity, purification of expressed protein and SDS-PAGE analysis. Conclusions From the current study, two conclusions came in picture, habitat is prime factor that regulation of chitin gene expression among many bacterial strains, second, moreover among the vibrio pathogenic strains (CT+) expression of chitinase gene is more precisely regulated by CT gene rather than external environments while in non-pathogenic strain ( CT-) completely absent.

Verma, Yogendra Kumar; Verma, Mahendra Kumar

2013-01-01

332

Responses of diploid and triploid Pacific oysters Crassostrea gigas to Vibrio infection in relation to their reproductive status.  

PubMed

Several Vibrio species are known to be pathogenic to the Pacific oyster Crassostrea gigas. Survival varies according to pathogen exposure and high mortality events usually occur in summer during gametogenesis. In order to study the effects of gametogenetic status and ploidy (a factor known to affect reproduction allocation in oysters) on vibriosis survival, we conducted two successive experiments. Our results demonstrate that a common bath challenge with pathogenic Vibrio splendidus and Vibrio aestuarianus on a mixture of mature, spawning and non-mature oysters can lead to significant mortality. Previous bath challenges, which were done using only non-mature oysters, had not produced mortality. Immunohistochemical analyses showed the affinity of Vibrio for gonadic tissues, highlighting the importance of sexual maturity for vibriosis infection processes in oysters. Mortality rate results showed poor repeatability between tanks, however, in this bath challenge. We then tested a standardized and repeatable injection protocol using two different doses of the same combination of two Vibrio species on related diploid and triploid oysters at four different times over a year. Statistical analyses of mortality kinetics over a 6-day period after injection revealed that active gametogenesis periods correspond to higher susceptibility to vibriosis and that there is a significant interaction of this seasonal effect with ploidy. However, no significant advantage of triploidy was observed. Triploid oysters even showed lower survival than diploid counterparts in winter. Results are discussed in relation to differing energy allocation patterns between diploid and triploid Pacific oysters. PMID:20833182

De Decker, S; Normand, J; Saulnier, D; Pernet, F; Castagnet, S; Boudry, P

2011-02-01

333

Predicting the Distribution of Vibrio spp. in the Chesapeake Bay: A Vibrio cholerae Case Study  

Microsoft Academic Search

Vibrio cholerae, the causative agent of cholera, is a naturally occurring inhabitant of the Chesapeake Bay and serves as a predictor for\\u000a other clinically important vibrios, including Vibrio parahaemolyticus and Vibrio vulnificus. A system was constructed to predict the likelihood of the presence of V. cholerae in surface waters of the Chesapeake Bay, with the goal to provide forecasts of the

Guillaume Constantin de Magny; Wen Long; Christopher W. Brown; Raleigh R. Hood; Anwar Huq; Raghu Murtugudde; Rita R. Colwell

2009-01-01

334

Validation of a method for the detection of five species, serogroups, biotypes and virulence factors of Vibrio by multiplex PCR in fish and seafood.  

PubMed

In this work a sequential multiplex PCR system was designed and validated for the detection of most frequent foodborne pathogen Vibrio species in fish and seafood (Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginoliticus and Vibrio mimicus). The method proposed functions in a hierarchical way, being composed of an end-point multiplex PCR to detect the presence of DNA belonging to the studied species, followed by multiplex PCR and fragment analysis allowing the viability assessment of the detected strains. The final multiplex PCR step of the method may be applied if identification of the serogroup, biotype and/or virulence factor level is necessary. Forty samples of commercial fish and seafood products were used at the method validation stage. Sixty three marine organism samples obtained from various estuarine areas of Spain including shrimps, crabs, bivalve mollusks and fishes were screened for presence of Vibrio species and 2 mussel samples were found positive for V. parahaemolyticus. On the whole, the proposed method is robust and readily adaptable in routine molecular diagnostic laboratories, allowing monitoring and simultaneous detection of all these bacterial pathogens in seafood samples, reducing the expenses and time consumed by other analytical methods. PMID:19913702

Espiñeira, Montserrat; Atanassova, Miroslava; Vieites, Juan M; Santaclara, Francisco J

2010-02-01

335

Evaluation of a rapid polymerase chain reaction based identification technique for Vibrio cholerae isolates.  

PubMed

Rapid and accurate identification of waterborne pathogens, such as Vibrio cholerae, in drinking-water sources is important to enable effective resource management and public health protection. Phenotypic systems currently being used for the identification of Vibrio cholerae isolates are time-consuming and the need exists for the development of suitable molecular techniques that can offer both fast and reliable identification. During this study, isolates identified as Vibrio cholerae by means of two different biochemical test systems (API 20E and VITEK 32) were analysed with the polymerase chain reaction (PCR) to compare the reliability of the various identification systems. The selected PCR technique amplified a sequence within the outer membrane protein of Vibrio cholerae, a gene specific for V. cholerae. It was found that out of 243 isolates biochemically identified as V. cholerae with either the API or VITEK system, 21 isolates did not give a positive result with the PCR detection method. Sequencing the 16S rDNA of more than half of these isolates and comparison of the sequences with Internet databases indicated that most of the isolates belonged to the genus Aeromonas. The results indicated that the rapid PCR procedure was more accurate than the API or VITEK systems currently being used for the phenotypic identification of Vibrio cholerae isolates. PMID:15318514

le Roux, W J; Masoabi, D; de Wet, C M E; Venter, S N

2004-01-01

336

The mechanoelectrical response of the cytoplasmic membrane of Vibrio cholerae  

PubMed Central

Persistence of Vibrio cholerae in waters of fluctuating salinity relies on the capacity of this facultative enteric pathogen to adapt to varying osmotic conditions. In an event of osmotic downshift, osmolytes accumulated inside the bacterium can be quickly released through tension-activated channels. With the newly established procedure of giant spheroplast preparation from V. cholerae, we performed the first patch-clamp characterization of its cytoplasmic membrane and compared tension-activated currents with those in Esherichia coli. Saturating pressure ramps revealed two waves of activation belonging to the ?1-nS mechanosensitive channel of small conductance (MscS)-like channels and ?3-nS mechanosensitive channel of large conductance (MscL)-like channels, with a pressure midpoint ratio p0.5MscS/p0.5MscL of 0.48. We found that MscL-like channels in V. cholerae present at a density three times higher than in E. coli, and yet, these vibrios were less tolerant to large osmotic downshocks. The Vibrio MscS-like channels exhibit characteristic inward rectification and subconductive states at depolarizing voltages; they also adapt and inactivate at subsaturating tensions and recover within 2 s upon tension release, just like E. coli MscS. Trehalose, a compatible internal osmolyte accumulated under hypertonic conditions, significantly shifts activation curves of both MscL- and MscS-like channels toward higher tensions, yet does not freely partition into the channel pore. Direct electrophysiology of V. cholerae offers new avenues for the in situ analysis of membrane components critical for osmotic survival and electrogenic transport in this pathogen.

Rowe, Ian; Elahi, Merina; Huq, Anwar

2013-01-01

337

Whole Transcriptome Profiling of Successful Immune Response to Vibrio Infections in the Oyster Crassostrea gigas by Digital Gene Expression Analysis  

PubMed Central

The cultivated Pacific oyster Crassostrea gigas has suffered for decades large scale summer mortality phenomenon resulting from the interaction between the environment parameters, the oyster physiological and/or genetic status and the presence of pathogenic microorganisms including Vibrio species. To obtain a general picture of the molecular mechanisms implicated in C. gigas immune responsiveness to circumvent Vibrio infections, we have developed the first deep sequencing study of the transcriptome of hemocytes, the immunocompetent cells. Using Digital Gene Expression (DGE), we generated a transcript catalog of up-regulated genes from oysters surviving infection with virulent Vibrio strains (Vibrio splendidus LGP32 and V. aestuarianus LPi 02/41) compared to an avirulent one, V. tasmaniensis LMG 20012T. For that an original experimental infection protocol was developed in which only animals that were able to survive infections were considered for the DGE approach. We report the identification of cellular and immune functions that characterize the oyster capability to survive pathogenic Vibrio infections. Functional annotations highlight genes related to signal transduction of immune response, cell adhesion and communication as well as cellular processes and defence mechanisms of phagocytosis, actin cytosqueleton reorganization, cell trafficking and autophagy, but also antioxidant and anti-apoptotic reactions. In addition, quantitative PCR analysis reveals the first identification of pathogen-specific signatures in oyster gene regulation, which opens the way for in depth molecular studies of oyster-pathogen interaction and pathogenesis. This work is a prerequisite for the identification of those physiological traits controlling oyster capacity to survive a Vibrio infection and, subsequently, for a better understanding of the phenomenon of summer mortality.

de Lorgeril, Julien; Zenagui, Reda; Rosa, Rafael D.; Piquemal, David; Bachere, Evelyne

2011-01-01

338

Whole transcriptome profiling of successful immune response to Vibrio infections in the oyster Crassostrea gigas by digital gene expression analysis.  

PubMed

The cultivated Pacific oyster Crassostrea gigas has suffered for decades large scale summer mortality phenomenon resulting from the interaction between the environment parameters, the oyster physiological and/or genetic status and the presence of pathogenic microorganisms including Vibrio species. To obtain a general picture of the molecular mechanisms implicated in C. gigas immune responsiveness to circumvent Vibrio infections, we have developed the first deep sequencing study of the transcriptome of hemocytes, the immunocompetent cells. Using Digital Gene Expression (DGE), we generated a transcript catalog of up-regulated genes from oysters surviving infection with virulent Vibrio strains (Vibrio splendidus LGP32 and V. aestuarianus LPi 02/41) compared to an avirulent one, V. tasmaniensis LMG 20012(T). For that an original experimental infection protocol was developed in which only animals that were able to survive infections were considered for the DGE approach. We report the identification of cellular and immune functions that characterize the oyster capability to survive pathogenic Vibrio infections. Functional annotations highlight genes related to signal transduction of immune response, cell adhesion and communication as well as cellular processes and defence mechanisms of phagocytosis, actin cytosqueleton reorganization, cell trafficking and autophagy, but also antioxidant and anti-apoptotic reactions. In addition, quantitative PCR analysis reveals the first identification of pathogen-specific signatures in oyster gene regulation, which opens the way for in depth molecular studies of oyster-pathogen interaction and pathogenesis. This work is a prerequisite for the identification of those physiological traits controlling oyster capacity to survive a Vibrio infection and, subsequently, for a better understanding of the phenomenon of summer mortality. PMID:21829707

de Lorgeril, Julien; Zenagui, Reda; Rosa, Rafael D; Piquemal, David; Bachère, Evelyne

2011-01-01

339

Recombination shapes the structure of an environmental Vibrio cholerae population.  

PubMed

Vibrio cholerae consists of pathogenic strains that cause sporadic gastrointestinal illness or epidemic cholera disease and nonpathogenic strains that grow and persist in coastal aquatic ecosystems. Previous studies of disease-causing strains have shown V. cholerae to be a primarily clonal bacterial species, but isolates analyzed have been strongly biased toward pathogenic genotypes, while representing only a small sample of the vast diversity in environmental strains. In this study, we characterized homologous recombination and structure among 152 environmental V. cholerae isolates and 13 other putative Vibrio isolates from coastal waters and sediments in central California, as well as four clinical V. cholerae isolates, using multilocus sequence analysis of seven housekeeping genes. Recombinant regions were identified by at least three detection methods in 72% of our V. cholerae isolates. Despite frequent recombination, significant linkage disequilibrium was still detected among the V. cholerae sequence types. Incongruent but nonrandom associations were observed for maximum likelihood topologies from the individual loci. Overall, our estimated recombination rate in V. cholerae of 6.5 times the mutation rate is similar to those of other sexual bacteria and appears frequently enough to restrict selection from purging much of the neutral intraspecies diversity. These data suggest that frequent recombination among V. cholerae may hinder the identification of ecotypes in this bacterioplankton population. PMID:21075874

Keymer, Daniel P; Boehm, Alexandria B

2011-01-01

340

Recombination Shapes the Structure of an Environmental Vibrio cholerae Population ? †  

PubMed Central

Vibrio cholerae consists of pathogenic strains that cause sporadic gastrointestinal illness or epidemic cholera disease and nonpathogenic strains that grow and persist in coastal aquatic ecosystems. Previous studies of disease-causing strains have shown V. cholerae to be a primarily clonal bacterial species, but isolates analyzed have been strongly biased toward pathogenic genotypes, while representing only a small sample of the vast diversity in environmental strains. In this study, we characterized homologous recombination and structure among 152 environmental V. cholerae isolates and 13 other putative Vibrio isolates from coastal waters and sediments in central California, as well as four clinical V. cholerae isolates, using multilocus sequence analysis of seven housekeeping genes. Recombinant regions were identified by at least three detection methods in 72% of our V. cholerae isolates. Despite frequent recombination, significant linkage disequilibrium was still detected among the V. cholerae sequence types. Incongruent but nonrandom associations were observed for maximum likelihood topologies from the individual loci. Overall, our estimated recombination rate in V. cholerae of 6.5 times the mutation rate is similar to those of other sexual bacteria and appears frequently enough to restrict selection from purging much of the neutral intraspecies diversity. These data suggest that frequent recombination among V. cholerae may hinder the identification of ecotypes in this bacterioplankton population.

Keymer, Daniel P.; Boehm, Alexandria B.

2011-01-01

341

Dynamics in genome evolution of Vibrio cholerae.  

PubMed

Vibrio cholerae, the etiological agent of the acute secretary diarrheal disease cholera, is still a major public health concern in developing countries. In former centuries cholera was a permanent threat even to the highly developed populations of Europe, North America, and the northern part of Asia. Extensive studies on the cholera bug over more than a century have made significant advances in our understanding of the disease and ways of treating patients. V. cholerae has more than 200 serogroups, but only few serogroups have caused disease on a worldwide scale. Until the present, the evolutionary relationship of these pandemic causing serogroups was not clear. In the last decades, we have witnessed a shift involving genetically and phenotypically varied pandemic clones of V. cholerae in Asia and Africa. The exponential knowledge on the genome of several representatives V. cholerae strains has been used to identify and analyze the key determinants for rapid evolution of cholera pathogen. Recent comparative genomic studies have identified the presence of various integrative mobile genetic elements (IMGEs) in V. cholerae genome, which can be used as a marker of differentiation of all seventh pandemic clones with very similar core genome. This review attempts to bring together some of the important researches in recent times that have contributed towards understanding the genetics, epidemiology and evolution of toxigenic V. cholerae strains. PMID:24462909

Banerjee, Rachana; Das, Bhabatosh; Balakrish Nair, G; Basak, Surajit

2014-04-01

342

Radiofrequency transmission line for bioluminescent Vibrio sp. irradiation  

NASA Astrophysics Data System (ADS)

We present the study and the analyses of a transmission line for radiofrequency (RF) irradiation of bacteria belonging to Vibrio harveyi-related strain PS1, a bioluminescent bacterium living in symbiosis with many marine organisms. The bioluminescence represents a new biologic indicator which is useful for studying the behaviour of living samples in the presence of RF waves due to the modern communication systems. A suitable transmission line, used as an irradiating cell and tested up to the maximum frequency used by the global system for mobile communications and universal mobile telecommunications system transmissions, was characterized. In this experiment, the RF voltage applied to the transmission line was 1 V. Due to short dimensions of the line and the applied high frequencies, standing waves were produced in addition to progressing waves and the electric field strength varies particularly along the longitudinal direction. The magnetic field map was not strongly linked to the electric one due to the presence of standing waves and of the outgoing irradiation. RF fields were measured by two homemade suitable probes able to diagnostic fields of high frequency. The field measurements were performed without any specimens inside the line. Being our sample made of living matter, the real field was modified and its value was estimated by a simulation code. The bioluminescence experiments were performed only at 900 MHz for two different measured electric fields, 53 and 140 V/m. The light emission was measured right from the beginning and after 7 and 25 h. Under RF irradiation, we found that the bioluminescence activity decreased. Compared with the control sample, the diminution was 6.8% and 44% after 7 and 25 h of irradiation, respectively, both with the low or high field. No changes of the survival factor for all the samples were observed. Besides, to understand the emission processes, we operated the deconvolution of the spectra by two Gaussian curves. The Gaussian peaks were approximately centered at 460 nm and 490 nm. The 490 nm peak was higher than the control one. Under RF, the 490 nm peak decreased compared to the 460 nm one. The decreasing was stronger for the sample in the higher field. The ratio of the emission area of the 490 nm to 460 nm was 5 for the control sample. It decreased up to 1.6 for the samples under RF. The bioluminescence improves the DNA repair by photoreactivation, and there is evidence that photolyase is preferentially activated by blue/violet light. Our finding suggests that RF exposure may stimulate DNA repair by shifting the emission spectra from blue/green (490 nm) to blue/violet (460 nm).

Nassisi, V.; Alifano, P.; Talà, A.; Velardi, L.

2012-07-01

343

Draft Genome Sequence of trh+ Vibrio parahaemolyticus VP-49, Isolated from Seafood Harvested along the Mangalore Coast, India  

PubMed Central

Vibrio parahaemolyticus is a seafood-borne pathogen autochthonous to the marine and estuarine ecosystem, which is responsible for gastroenteritis due to the consumption of contaminated raw seafood. Here, we report the draft genome sequence of V. parahaemolyticus VP-49, isolated from seafood, to identify the different virulence attributes and to study the mechanisms that enhance its environmental fitness.

Kumar, Ballamoole Krishna; Deekshit, Vijaya Kumar; Rai, Praveen; Gurtler, Volker; Karunasagar, Iddya

2014-01-01

344

A Common Virulence Plasmid in Biotype 2 Vibrio vulnificus and Its Dissemination Aided by a Conjugal Plasmid  

Microsoft Academic Search

Strains of Vibrio vulnificus, a marine bacterial species pathogenic for humans and eels, are divided into three biotypes, and those virulent for eels are classified as biotype 2. All biotype 2 strains possess one or more plasmids, which have been shown to harbor the biotype 2-specific DNA sequences. In this study we determined the DNA sequences of three biotype 2

Chung-Te Lee; Carmen Amaro; Keh-Ming Wu; Esmeralda Valiente; Yi-Feng Chang; Shih-Feng Tsai; Chuan-Hsiung Chang; Lien-I Hor

2008-01-01

345

Identification and Characterization of OscR, a Transcriptional Regulator Involved in Osmolarity Adaptation in Vibrio cholerae  

Microsoft Academic Search

Vibrio cholerae is a facultative human pathogen. In its aquatic habitat and as it passes through the digestive tract, V. cholerae must cope with fluctuations in salinity. We analyzed the genome-wide transcriptional profile of V. cholerae grown at different NaCl concentrations and determined that the expression of compatible solute biosynthesis and transporter genes, virulence genes, and genes involved in adhesion

Nicholas J. Shikuma; Fitnat H. Yildiz

2009-01-01

346

Isolation and Characterization of Vibrio tubiashii Outer Membrane Proteins and Determination of a toxR Homolog  

Microsoft Academic Search

Although Vibrio tubiashii was originally found to cause bacillary necrosis in larval and juvenile mollusks (3, 10, 27, 28), recent studies have shown that it can also cause diarrhea in suckling mice (8). Additionally, its ability to cause death in fish (1) has led to the conclusion that V. tubiashii may also be a finfish pathogen. In the present study,

J. Jean-Gilles Beaubrun; M. H. Kothary; S. K. Curtis; N. C. Flores; B. E. Eribo; B. D. Tall

2008-01-01

347

Multiplex PCR Assay for Detection of Vibrio vulnificus Biotype 2 and Simultaneous Discrimination of Serovar E Strains?  

PubMed Central

In the present work we develop a multiplex PCR assay for the detection and identification of the fish pathogen Vibrio vulnificus biotype 2 with discriminating potential for zoonotic strains (serovar E). The PCR assay allowed the identification of two new biotype 2 serovar E human isolates from culture collections. Finally, the multiplex was successfully applied to both diagnosis and carrier detection in field samples.

Sanjuan, Eva; Amaro, Carmen

2007-01-01

348

RAPID TETRAZOLIUM DYE REDUCTION ASSAY TO ASSESS THE BACTERICIDAL ACTIVITY OF OYSTER (CRASSOSTREA VIRGINICA) HEMOCYTES AGAINST VIBRIO PARAHAEMOLYTICUS  

EPA Science Inventory

An assay was developed to assess the ability of oyster, Crassostrea virginica, hemocytes to kill the human pathogenic bacterium, Vibrio parahaemolyticus (ATCC 17802). Bacterial killing was estimated colorimetrically by the enzymatic reduction of a tetrazolium dye, 3-(4,5-dimethyl...

349

Growth, luminescence, respiration, and the ATP pool during autoinduction in Beneckea harveyi.  

PubMed Central

The bacterial bioluminescence system is unusual because it is self-induced. In the late logarithmic phase of growth, upon the accumulation of an autoinducer, the synthesis of the components of the system is initiated. We were interested in determining what effect this burst of synthesis and activity has on cellular energy metabolism. The ATP pool of the luminous bacterium Beneckea harveyi was found to dip 10- to 20-fold during the luminescence period, while the respiration per unit cell mass (optical density) increased but by much less. The dip in the ATP pool did not occur in four different types of dark mutants, including one that was temperature conditional and another that was conditional upon added cyclic AMP for luminescence. However, it is neither the synthesis nor the activity of luciferase that is responsible for the ATP dip; the dip does not occur in certain dark "aldehyde" mutants which nevertheless synthesize normal levels of luciferase, whereas it does occur at 36 degrees C in a temperature-sensitive luciferase mutant which forms normal levels of inactive luciferase. Results with other aldehyde mutants implicate the pathway involved in the synthesis of the aldehyde factor with the ATP dip.

Ulitzur, S; Hastings, J W

1978-01-01

350

Survey of postharvest-processed oysters in the United States for levels of Vibrio vulnificus and Vibrio parahaemolyticus.  

PubMed

From June through October 2004, the U.S. Food and Drug Administration collected oysters (61 samples) that had been subjected to postharvest processing (PHP) methods, including mild heat treatment, freezing, and high hydrostatic pressure, from processors and retail markets in various states to determine Vibrio vulnificus and V. parahaemolyticus levels. Presence in a 25-g sample and most probable number (MPN) using standard enrichment and selective isolation procedures were utilized. Suspect colonies were isolated and identified using DNA probe colony hybridization. Neither species of vibrio was detected in 25-g portions of most samples regardless of the PHP. The lowest frequency of isolation of either pathogen (<10%) was observed with the mild heat process. Few (12 to 13%) frozen samples collected at the processor but not at retail contained >30 MPN/g of either pathogen. The mean levels of either organism in PHP oysters observed in the present study were 5 to 6 log less than in unprocessed raw Gulf Coast oysters. Of the 70 V. vulnificus isolates examined, only 5 possessed the putative virulence marker, type B 16S rRNA. Neither the thermostable direct hemolysin (tdh) nor the tdh-related hemolysin (trh) virulence gene was detected in any of the 40 V. parahaemolyticus isolates examined in the present study. These data suggest that if there is any selective advantage to pathogenic strains of V. vulnificus and V. parahaemolyticus, these differences are minimal. These results indicate that all PHP treatments greatly reduce exposure of V. vulnificus and V. parahaemolyticus to raw-oyster consumers. Consequently, these PHP oysters pose a much lower risk of illness to consumers due to these pathogens. PMID:19833034

Depaola, Angelo; Jones, Jessica L; Noe, Kathy E; Byars, Robin H; Bowers, John C

2009-10-01

351

The ability of Vibrio vulnificus to use a synthetic hydrophilic heme compound, Fe-TPPS, as a single iron source  

Microsoft Academic Search

Vibrio vulnificus, an opportunistic human pathogen, can obtain iron from a variety of heme proteins. This process involves the digestion of heme proteins by an exoprotease to liberate protoheme (iron-protoporphyrin IX). In the present study, we tested whether this pathogen also uses a synthetic heme compound, Fe-?,?,?,?-tetraphenylporphine tetrasulfonic acid (Fe-TPPS), as an iron source. When inoculated into a medium containing

Shin-ichi Miyoshi; Takehito Kamei; Yu Inami; Yuki Ota; Shigeo Yamamoto; Ken-ichi Tomochika; Sumio Shinoda

1999-01-01

352

Multiplex PCR assays for the detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae with an internal amplification control.  

PubMed

A multiplex polymerase chain reaction (PCR) assay that can simultaneously detect 4 major Vibrio spp., Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae, in the presence of an internal amplification control (IAC) was developed. Species-specific PCR primers were designed based on the gyrB gene for V. alginolyticus, the collagenase gene for V. parahaemolyticus, the vvhA gene for V. vulnificus, and the ompW gene for V. cholerae. Additionally, an IAC primer pair was designed in conserved regions of the bacterial 16S rRNA gene that is used to indicate false-negative results. A multiplex PCR method was developed after optimization of the reaction conditions. The specificity of the PCR was validated by using 83 Vibrio strains and 10 other non-Vibrio bacterial species. The detection limit of the PCR was 10 CFU per tube for V. alginolyticus, V. parahaemolyticus, V. vulnificus, and 10(5) CFU per tube for V. cholerae in mixed conditions. This method was used to identify 69 suspicious Vibrio isolates, and the results were consistent with physiological and biochemical tests. This multiplex PCR method proved to be rapid, sensitive, and specific. The existence of IAC could successfully eliminate false-negative results for the detection of V. alginolyticus, V. parahaemolyticus, V. vulnificus, and V. cholerae. PMID:24731836

Wei, Shuang; Zhao, Hui; Xian, Yuyin; Hussain, Malik A; Wu, Xiyang

2014-06-01

353

Quorum-sensing regulators control virulence gene expression in Vibrio cholerae  

Microsoft Academic Search

The production of virulence factors including cholera toxin and the toxin-coregulated pilus in the human pathogen Vibrio cholerae is strongly influenced by environmental conditions. The well-characterized ToxR signal transduction cascade is responsible for sensing and integrating the environmental information and controlling the virulence regulon. We show here that, in addition to the known components of the ToxR signaling circuit, quorum-sensing

Jun Zhu; Melissa B. Miller; Russell E. Vance; Michelle Dziejman; John J. Mekalanos

2002-01-01

354

Survival and viability of nonculturable Escherichia coli and Vibrio cholerae in the estuarine and marine environment  

Microsoft Academic Search

Plating methods for estimating survival of indicator organisms, such asEscherichia coli, and water-borne pathogens includingVibrio cholerae, have severe limitations when used to estimate viable populations of these organisms in the aquatic environment. By combining the methods of immunofluorescent microscopy, acridine orange direct counting, and direct viable counting, with culture methods such as indirect enumeration by most probable number (MPN) estimation

Huai-Shu Xu; N. Roberts; F. L. Singleton; R. W. Attwell; D. J. Grimes; R. R. Colwell

1982-01-01

355

Vibrio Zinc-Metalloprotease Causes Photoinactivation of Coral Endosymbionts and Coral Tissue Lesions  

Microsoft Academic Search

BackgroundCoral diseases are emerging as a serious threat to coral reefs worldwide. Of nine coral infectious diseases, whose pathogens have been characterized, six are caused by agents from the family Vibrionacae, raising questions as to their origin and role in coral disease aetiology.Methodology\\/Principal FindingsHere we report on a Vibrio zinc-metalloprotease causing rapid photoinactivation of susceptible Symbiodinium endosymbionts followed by lesions

Meir Sussman; Jos C. Mieog; Jason Doyle; Steven Victor; Bette L. Willis; David G. Bourne; John F. Bruno

2009-01-01

356

Detection of viable Vibrio cholerae by reverse-transcriptase polymerase chain reaction (RT-PCR)  

Microsoft Academic Search

The use of conventional PCR can amplify target DNA from both viable and nonviable cells ofVibrio cholera. Detection of only viable microbial pathogens in biological samples, especially clinical and food samples, is usually desired\\u000a to ensure positive test results are associated with active agents, and not the remains of dead cells. Positive identifications\\u000a caused by nonliving causative agents may lead

Asim K. Bej; Wee-Yao Ng; Shellie Morgan; Daniel D. Jones; Meena H. Mahbubani

1996-01-01

357

Genetic characterization of Vibrio cholerae strains by inter simple sequence repeat-PCR  

Microsoft Academic Search

The utility of inter simple sequence repeat-PCR (ISSR-PCR) assay in the characterization and elucidation of the phylogenetic relationship between the pathogenic and nonpathogenic isolates of Vibrio cholerae is demonstrated. A total of 45 V. cholerae strains including 15 O1 El Tor, nine O139 and 21 non-O1\\/non- O139 strains were analyzed using eight ISSR primers. These primers, which are essentially simple

A. Ravi Kumar; V. Sathish; G. Balakrish Nair; J. Nagaraju

2007-01-01

358

The Amazonia Variant of Vibrio cholerae: Molecular Identification and Study of Virulence Genes  

Microsoft Academic Search

The pathogenic O1 Amazonia variant of Vibrio cholerae has been shown previously to have a cytotoxin acting on cultured Vero and Y-1 cells, and to lack important virulence factors such as the cholera toxin (Coelho et al. 1995a). This study extends the molecular analysis of the Amazonia strains, detecting the pres- ence of the toxR gene, with a very similar

MAS Baptista; JRC Andrade; ACP Vicente; CA Salles; A Coelho

1998-01-01

359

Rapid Screening of Toxigenic Vibrio cholerae O1 Strains from South Iran by PCR-ELISA  

Microsoft Academic Search

Background: The ability to sensitively detect Vibrio cholera with PCR-ELISA method represents a considerable advancement over alternative more time-consuming methods for detection of this pathogen. The aim of this research is to evaluate the suitability of a PCR-enzyme-linked immunosorbent assay for sensitive and rapid detection of V. cholera O1. Methods: The 398-bp sequence of a gene that codes for the

Seyed Latif Mousavi; Shahram Nazarian; Jafar Amani; Ahmad Karimi Rahgerdi

2008-01-01

360

csrA inhibits the formation of biofilms by Vibrio vulnificus.  

PubMed

PCR screening of the shellfish-borne pathogen Vibrio vulnificus revealed csrA-negative strains, and these strains formed increased biofilm compared to csrA-positive strains. Complementation in trans with csrA resulted in reduced biofilm formation, similar to that by csrA(+) strains. Our results provide evidence that csrA inhibits biofilm formation in V. vulnificus. PMID:18820071

Jones, Melissa K; Warner, Elizabeth B; Oliver, James D

2008-11-01

361

A Potential Epidemic Factor from the Bacteria, Vibrio cholerae WO7  

Microsoft Academic Search

Certain species of Vibrio cholerae have evolved mechanisms to become pathogenic to humans, with the potential to cause a severe life-threatening diarrheal disease,\\u000a cholera. Cholera can emerge as explosive outbreaks in the human population. V. cholerae illness is produced primarily through the expression of a potent toxin (cholera toxin) within the human intestine. The present\\u000a study has been carried out

Shalmoli Bhattacharyya; Jasleen Shant; Nirmal K. Ganguly; Siddhartha Majumdar; Sujata Ghosh

2008-01-01

362

NspS, a Predicted Polyamine Sensor, Mediates Activation of Vibrio cholerae Biofilm Formation by Norspermidine  

Microsoft Academic Search

Vibrio cholerae is both an environmental bacterium and a human intestinal pathogen. The attachment of bacteria to surfaces in biofilms is thought to be an important feature of the survival of this bacterium both in the environment and within the human host. Biofilm formation occurs when cell-surface and cell-cell contacts are formed to make a three-dimensional structure characterized by pillars

Ece Karatan; Tammi R. Duncan; Paula I. Watnick

2005-01-01

363

Characterization of Enhancer Binding by the Vibrio cholerae Flagellar Regulatory Protein FlrC  

Microsoft Academic Search

The human pathogen Vibrio cholerae is a highly motile organism by virtue of a polar flagellum, and motility has been inferred to be an important aspect of virulence. It has previously been demonstrated that the 54-dependent activator FlrC is necessary for both flagellar synthesis and for enhanced intestinal colonization. In order to characterize FlrC binding, we analyzed two FlrC-dependent promoters,

Nidia E. Correa; Karl E. Klose

2005-01-01

364

Pathogen-pathogen interaction  

PubMed Central

There is growing awareness of the health implications of the fact that infectious agents often do not act independently; rather their disease potential is mediated in diverse and significant ways by their relationships with other pathogens. Pathogen-pathogen interaction (PPI), for example, impacts various virulence factors in human infection. Although still in its infancy, the study of PPI, a form of epidemiological synergism, is emerging as an important arena of new research and new understanding in health and clinical care. The aims of this paper are to: (1) draw attention to the role of PPI in human disease patterns; (2) present the syndemics model as a biosocial approach for examining the nature, pathways, contexts, and health implications of PPI and (3) suggest the utility of this approach to PPI. Toward these ends, this paper (a) reviews three case examples of alternative PPIs, (b) describes the development and key concepts and components of the syndemics model with specific reference to interacting infectious agents, (c) contextualizes this discussion with a brief review of broader syndemics disease processes (not necessarily involving infections disease) and (d) comments on the research, treatment and prevention implications of syndemic interaction among pathogens.

2010-01-01

365

Prevalence and antimicrobial susceptibilities of Vibrio, salmonella, and Aeromonas isolates from various uncooked seafoods in Thailand.  

PubMed

Uncooked seafood samples were collected from open markets and supermarkets in Bangkok, Thailand, and were examined for the presence of Vibrio, Salmonella, and Aeromonas species from January to February 2008. From 120 samples, 272 bacterial isolates were identified through biochemical testing. Of all sea bass, shrimp, oyster, and blood cockle samples (30 of each) that were processed for culture, 114 (95%) samples had at least one detectable isolate of Vibrio, Salmonella, or Aeromonas, leaving only 6 (5%) samples free of them. All oyster sample (100%) had at least one pathogen, followed by sea bass (97%), blood cockles (97%), and shrimp (90%). Overall, 111 (92%) of all samples had detectable Vibrio spp., 32 (27%) had detectable Aeromonas spp., and 25 (21%) had detectable Salmonella enterica. There was no overall difference between positive samples collected from fresh markets versus supermarkets (relative risk, 0.97; 95% CI, 0.89 to 1.05). Resistance to ampicillin among isolated pathogens was relatively high (56%), while resistance to 12 other antibiotics, including azithromycin, ciprofloxacin, and trimethoprim-sulfamethoxazole, was relatively low (0, 0, and 3%, respectively). Study results indicate that uncooked seafood in Bangkok, Thailand, commonly harbors enteric pathogens and that consumption of uncooked seafood should be avoided to reduce foodborne illnesses. PMID:22221354

Woodring, Joseph; Srijan, Apichai; Puripunyakom, Paksathorn; Oransathid, Wilawan; Wongstitwilairoong, Boonchai; Mason, Carl

2012-01-01

366

Rapid simultaneous screening of seven clinically important enteric pathogens using a magnetic bead based DNA microarray  

Microsoft Academic Search

In this study, we describe a DNA microarray assay by using bead-mediated visible light-assisted signal detection for simultaneous\\u000a screening of seven clinically important enteric pathogens, including Escherichia coli O157:H7, Vibrio cholerae, Vibrio parahaemolyticus, Salmonella spp., Staphylococcus aureus, Rotavirus, and Norwalk virus (including genogroup I and II). Seven pairs of primers, in which the forward primers were labeled with biotin at

Hong Sun; Qiu-Hua Mo; Ji-Can Lin; Ze Yang; Cheng-Ning Tu; Da-Yong Gu; Lei Shi; Wei-Ping Lu

2011-01-01

367

Molecular analysis of the emergence of pandemic Vibrio parahaemolyticus  

PubMed Central

Background Vibrio parahaemolyticus is abundant in the aquatic environment particularly in warmer waters and is the leading cause of seafood borne gastroenteritis worldwide. Prior to 1995, numerous V. parahaemolyticus serogroups were associated with disease, however, in that year an O3:K6 serogroup emerged in Southeast Asia causing large outbreaks and rapid hospitalizations. This new highly virulent strain is now globally disseminated. Results We performed a four-way BLAST analysis on the genome sequence of V. parahaemolyticus RIMD2210633, an O3:K6 isolate from Japan recovered in 1996, versus the genomes of four published Vibrio species and constructed genome BLAST atlases. We identified 24 regions, gaps in the genome atlas, of greater than 10 kb that were unique to RIMD2210633. These 24 regions included an integron, f237 phage, 2 type III secretion systems (T3SS), a type VI secretion system (T6SS) and 7 Vibrio parahaemolyticus genomic islands (VPaI-1 to VPaI-7). Comparative genomic analysis of our fifth genome, V. parahaemolyticus AQ3810, an O3:K6 isolate recovered in 1983, identified four regions unique to each V. parahaemolyticus strain. Interestingly, AQ3810 did not encode 8 of the 24 regions unique to RMID, including a T6SS, which suggests an additional virulence mechanism in RIMD2210633. The distribution of only the VPaI regions was highly variable among a collection of 42 isolates and phylogenetic analysis of these isolates show that these regions are confined to a pathogenic clade. Conclusion Our data show that there is considerable genomic flux in this species and that the new highly virulent clone arose from an O3:K6 isolate that acquired at least seven novel regions, which included both a T3SS and a T6SS.

Boyd, E Fidelma; Cohen, Ana Luisa V; Naughton, Lynn M; Ussery, David W; Binnewies, Tim T; Stine, O Colin; Parent, Michelle A

2008-01-01

368

Detection, Isolation, and Identification of Vibrio cholerae from the Environment  

PubMed Central

Recent molecular advances in microbiology have greatly improved the detection of bacterial pathogens in the environment. Improvement and a downward trend in the cost of molecular detection methods have contributed to increased frequency of detection of pathogenic microorganisms where traditional culture-based detection methods have failed. Culture methods also have been greatly improved and the confluence of the two suites of methods provides a powerful tool for detection, isolation, and characterization of pathogens. While molecular detection provides data on the presence and type of pathogens, culturing methods allow a researcher to preserve the organism of interest for “–omics” studies, such as genomic, metabolomic, secretomic, and transcriptomic analysis, which are rapidly becoming more affordable. This has yielded a clearer understanding of the ecology and epidemiology of microorganisms that cause disease. Specifically, important advances have been made over the past several years on isolation, detection, and identification of Vibrio cholerae, the causative agent of cholera in humans. In this unit, we present commonly accepted methods for isolation, detection, and characterization of V. cholerae, providing more extensive knowledge of the ecology and epidemiology of this organism. This unit has been fully revised and updated from the earlier unit (Huq, Grim et al. 2006) with the latest knowledge and additional information not previously included. We have also taken into account of cost of reagents and equipment that may be prohibitive for many researchers and have, therefore, included protocols for all laboratories, including those with limited resources, likely to be located in regions of cholera endemicity.

Huq, Anwar; Haley, Bradd J.; Taviani, Elisa; Chen, Arlene; Hasan, Nur A.; Colwell, Rita R.

2012-01-01

369

Isolation of Vibrio alginolyticus and Vibrio splendidus from captive-bred seahorses with disease symptoms.  

PubMed

Vibrio species isolated from diseased seahorses were characterized by PCR amplification of repetitive bacterial DNA elements (rep-PCR) and identified by 16S ribosomal RNA gene sequence analysis. The results demonstrated that Vibrio alginolyticus and Vibrio splendidus were predominant in the lesions of these seahorses. To our knowledge, this is the first time that these bacterial species have been associated with disease symptoms in captive-bred seahorses. PMID:19921544

Balcázar, José L; Gallo-Bueno, Alfonso; Planas, Miquel; Pintado, José

2010-02-01

370

Antimicrobial Susceptibilities of Vibrio parahaemolyticus and Vibrio vulnificus Isolates from Louisiana Gulf and Retail Raw Oysters?  

PubMed Central

The antimicrobial susceptibilities of 168 Vibrio parahaemolyticus and 151 Vibrio vulnificus isolates recovered from 82 Louisiana Gulf and retail oysters in 2005 and 2006 were determined. Overall, the two vibrios remained susceptible to the majority of antimicrobials tested; reduced susceptibility was detected only in V. parahaemolyticus for ampicillin (81%; MIC ? 16 ?g/ml). Additionally, V. parahaemolyticus displayed significantly higher MICs for cefotaxime, ciprofloxacin, and tetracycline than V. vulnificus.

Han, Feifei; Walker, Robert D.; Janes, Marlene E.; Prinyawiwatkul, Witoon; Ge, Beilei

2007-01-01

371

Molecular cloning of a C-type lectin with two CRD domains from the banana shrimp Fenneropenaeus merguiensis: Early gene up-regulation after Vibrio harveyi infection  

Microsoft Academic Search

A diverse class of pattern-recognition proteins called lectins play important roles in shrimp innate immunity. A novel C-type lectin gene (FmLC) was cloned from the hepatopancreas of banana shrimp Fenneropenaeus merguiensis by means of PCR and 5? and 3? rapid amplification of cDNA ends (RACE). The full-length cDNA consists of 1118bp with one 1002bp open reading frame, encoding 333 amino

Onnicha Rattanaporn; Prapaporn Utarabhand

2011-01-01

372

Phenotypic characterization of Vibrio vulnificus biotype 2, a lipopolysaccharide-based homogeneous O serogroup within Vibrio vulnificus.  

PubMed Central

In this study, we have reevaluated the taxonomic position of biotype 2 of Vibrio vulnificus. For this purpose, we have biochemically and serologically characterized 83 biotype 2 strains from diseased eels, comparing them with 17 biotype 1 strains from different sources. Selected strains were also molecularly analyzed and tested for eel and mouse pathogenicity. Results have shown that biotype 2 (i) is biochemically homogeneous, indole production being the main trait that distinguishes it from biotype 1, (ii) presents small variations in DNA restriction profiles and outer membrane protein patterns, some proteins being immunologically related to outer membrane proteins from biotype 1, (iii) expresses a common lipopolysaccharide (LPS) profile, which is immunologically identical among strains and distinct from that of LPS of tested biotype 1 strains, and (iv) contains at least two high-Mr plasmids. Regarding host range, we have confirmed that both biotypes are pathogenic for mice but only biotype 2 is pathogenic for eels. On the basis of these data, we propose that biotype 2 of V. vulnificus constitutes an LPS-based O serogroup which is phenotypically homogeneous and pathogenic for eels. In this article, the serogroup is designated serogroup E (for eels).

Biosca, E G; Oliver, J D; Amaro, C

1996-01-01

373

Detection and identification of enterohemorrhagic Escherichia coli O157:H7 and Vibrio cholerae O139 using oligonucleotide microarray  

PubMed Central

Background The rapid and accurate detection and identification of the new subtype of the pathogens is crucial for diagnosis, treatment and control of the contagious disease outbreak. Here, in this study, an approach to detect and identify Escherichia coli O157:H7 and Vibrio cholerae O139 was established using oligonucleotide microarray. We coupled multiplex PCR with oligonucleotide microarray to construct an assay suitable for simultaneous identification of two subtypes of the pathogens. Results The stx1, stx2 gene and uidA gene having the specific mutant spot were chosen as the targets for Escherichia coli O157:H7, and meanwhile the ctxA, tcpA, and LPSgt gene for Vibrio cholerae O139. The oligonucleotide microarray was composed of eight probes including negative control and positive control from 16S rDNA gene. The six primers were designed to amplify target fragments in two triplex PCR, and then hybridized with oligonucleotide microarray. An internal control would be to run a PCR reaction in parallel. Multiplex PCR did not produce any non-specific amplicons when 149 related species or genera of standard bacteria were tested (100% specificity). In addition, Escherichia coli O157:H7 and Escherichia coli O157:non-H7, Vibrio cholerae O139 and Vibrio cholerae O1 had been discriminated respectively. Using recombinant plasmid and target pathogens, we were able to detect positive hybridization signals with 102 copies/?L and 103 cfu/mL per reaction. Conclusion The DNA microarray assay reported here could detect and identify Escherichia coli O157:H7 and Vibrio cholerae O139, and furthermore the subtype was distinguished. This assay was a specific and sensitive tool for simultaneous detection and identification of the new subtype of two pathogens causing diarrhea in human.

Jin, Da-Zhi; Xu, Xiao-Jing; Chen, Su-Hong; Wen, Si-Yuan; Ma, Xue-En; Zhang, Zheng; Lin, Feng; Wang, Sheng-Qi

2007-01-01

374

Genome sequence of Vibrio splendidus: an abundant planctonic marine species with a large genotypic diversity.  

PubMed

Vibrio splendidus is a dominant Vibrio species in seawater presenting a remarkable genetic diversity; several strains have been linked to invertebrate's mortality. We report the complete genome sequence of V. splendidus LGP32, an oyster pathogen, and its comparison with partial genome sequences from related strains. As is typical for the genus, V. splendidus LGP32 contains two chromosomes (3.29 and 1.67 Mb) and most essential cellular processes are encoded by chromosome 1. Comparison with two other V. splendidus partial genome sequences (strains 12B01 and Med222) confirms the previously suggested high genotypic diversity within this species and led to the identification of numerous strain-specific regions that could frequently not be assigned to a specific mechanisms of recombination. Surprisingly, the chromosomal integron, the most variable genetic element in all other Vibrio species analysed to date, is absent from 12B01 and inactivated by a mobile element in Med222, while in LGP32 it only contains a limited number of cassettes. Finally, we found that the LGP32 integron contains a new dfrA cassette, related to those found in resistance integrons of gram-negative clinical isolates. Those results suggest that marine Vibrio can be a source of antibiotic resistance genes. PMID:19364337

Le Roux, Frédérique; Zouine, Mohamed; Chakroun, Nesrine; Binesse, Johan; Saulnier, Denis; Bouchier, Christiane; Zidane, Nora; Ma, Laurence; Rusniok, Christophe; Lajus, Aurélie; Buchrieser, Carmen; Médigue, Claudine; Polz, Martin F; Mazel, Didier

2009-08-01

375

A simple fluorogenic method to detect Vibrio cholerae and Aeromonas hydrophila in well water for areas impacted by catastrophic disasters.  

PubMed

The colony overlay procedure for peptidases (COPP) is a simple, fluorogenic assay that can rapidly detect and quantify Vibrio cholerae and Aeromonas hydrophila in well water. Cleavage of the substrate L-lysyl-7-amino-4-trifluoromethylcoumarin by enzymes present in Vibrio and Aeromonas species produces fluorescent foci on cellulose acetate membranes exposed to long-wave ultraviolet light. Vibrio cholerae O1, O139, O155, and A. hydrophila were readily detected using this procedure, whereas Enterobacteriaceae and other non-Vibrionaceae pathogens did not produce fluorescence. The assay is practical for assessing the relative safety of well water in areas that have experienced catastrophic devastation from natural disasters, acts of war, or civil strife and may help curb outbreaks of cholera and other enteric illnesses in affected areas. In tropical climates, the procedure may be adapted for use in areas without electricity. PMID:16968933

Richards, Gary P; Watson, Michael A

2006-09-01

376

Differential Expression of Vibrio vulnificus Capsular Polysaccharide  

PubMed Central

Vibrio vulnificus is a human pathogen whose virulence has been associated with the expression of capsular polysaccharide (CPS). Multiple CPS types have been described; however, virulence does not appear to correlate with a particular CPS composition. Reversible-phase variation for opaque and translucent colony morphologies is characterized by changes in CPS expression, as suggested by electron microscopy of cells stained nonspecifically with ruthenium red. Isolates with opaque colony morphologies are virulent and appear to be more thickly encapsulated than naturally occurring translucent-phase variants, which have reduced, patchy, or absent CPS. Previously, we have shown that the virulence of translucent-phase variants was intermediate between opaque-phase variants and acapsular transposon mutants, suggesting a correlation between virulence and the amount of CPS expressed. In the present study, CPS expression of phase variants and genetically defined mutants of V. vulnificus M06-24/O was examined by using a CPS-specific monoclonal antibody with an enzyme-linked immunosorbent assay, flow cytometry, and immunoelectron microscopy. Semiquantitative analyses of CPS expression correlated well among these assays, confirming that the translucent-phase variant was intermediate in CPS expression and retained type I CPS-specific epitopes. Cell surface expression of CPS varied with the growth phase, increasing during logarithmic growth and declining in stationary culture. Significantly greater CPS expression (P = 0.026) was observed for cells grown at 30°C than for those at 37°C. These studies confirm that phase variation and virulence in V. vulnificus correlate with the amount of CPS expressed and demonstrate the fluidity of bacterial polysaccharide expression in response to environmental conditions.

Wright, Anita C.; Powell, Jan L.; Tanner, Mike K.; Ensor, Lynne A.; Karpas, Arthur B.; Morris, J. Glenn; Sztein, Marcelo B.

1999-01-01

377

Vibrio kanaloae sp. nov., Vibrio pomeroyi sp. nov. and Vibrio chagasii sp. nov., from sea water and marine animals.  

PubMed

The taxonomic position of the fluorescent amplified fragment length polymorphism fingerprinting groups A46 (five isolates), A51 (six isolates), A52 (five isolates) and A53 (seven isolates) obtained in a previous study were further analysed through a polyphasic approach. The 23 isolates were phylogenetically related to Vibrio splendidus, but DNA-DNA hybridization experiments proved that they belong to three novel species. Chemotaxonomic and phenotypic analyses further disclosed several features that differentiate between the 23 isolates and known Vibrio species. The names Vibrio kanaloae sp. nov. (type strain LMG 20539(T) = CAIM 485(T); EMBL accession no. AJ316193; G + C content 44.7 mol%), Vibrio pomeroyi sp. nov. (type strain LMG 20537(T) = CAIM 578(T); EMBL accession no. AJ491290; G +C content 44.1 mol%) and Vibrio chagasii sp. nov. (type strain LMG 21353(T) = CAIM 431(T); EMBL accession no. AJ316199; G + C content 44.6 mol%) are respectively proposed to encompass the five isolates of A46, the six isolates of A51 and the 12 isolates of A52/A53. The three novel species can be distinguished from known Vibrio species by several phenotypic features, including utilization and fermentation of various carbon sources, beta-galactosidase activity and fatty acid content (particularly of 12 : 0, 14: 0, 14 : 0 iso and 16 : 0 iso). PMID:12807197

Thompson, F L; Thompson, C C; Li, Y; Gomez-Gil, B; Vandenberghe, J; Hoste, B; Swings, J

2003-05-01

378

[Classification and identification of vibrio cholerae and vibrio parahaemolyticus isolates based on gyrB gene phylogenetic analysis].  

PubMed

In order to validate the usefulness of gyrB genotype for the classification and identification of Vibrio cholerae and Vibrio parahaemolyticus isolates, the phylogenetic analysis of 13 V. cholerae, 8 V. parahaemolyticus, 2 Aeromonas hydrophila and 1 Plesiomonas shigelloides strains was carried out using the partial coding sequence of gyrB, a gene that encodes the B subunit of DNA gyrase (topoisomerase type II ) in bacteria. These strains were separately clustered at species level and typed by the DNA sequences of reference strains from GenBank. CtxA positive V. cholerae strains including 8 clincical isolates of 0139 and 2 clinical isolates of 01 formed one cluster. Four V. parahaemolyticus strains of 1 isolate from 2002 Zhejiang outbreak patient ( tdh positive), 2 clinical isoltates from 2004 and 1 strain from Japan were grouped with an environmental isolate ( trh positive) from 2001. GyrB genotype is applicable to species identification of V. cholerae, V. parahaemolyticus, A. hydrophila and P. shigelloides isolates. The ctxA positive 0139 and 01 group of V. cholerae are closely related, as reflected by gyrB sequence divergence. Furthermore, the toxigenic V. parahaemolyticus strain isolated from environments may be the potential pathogen to the local prevalent and sporadic cases. PMID:17302148

Hou, Xiao-li; Cao, Qing-yi; Pan, Jin-cao; Chen, Zhi

2006-12-01

379

Induction of Fimbriated Vibrio cholerae O139  

Microsoft Academic Search

Several fimbriated phases of Vibrio cholerae O139 strains were selectively induced and compared immuno- logically and biochemically with those of V. cholerae O1. Fimbrial antigens were detected on the surfaces of vibrio cells colonizing the epithelial cells of a rabbit small intestine. Convalescent-phase sera from six individuals infected with V. cholerae O139 revealed the development of antibody against the fimbrillin.

MASAHIKO EHARA; MAMORU IWAMI; YOSHIO ICHINOSE; TOSHIYA HIRAYAMA; M. JOHN ALBERT; R. BRADLEY SACK; SHOICHI SHIMODORI

1998-01-01

380

Vibrio alginolyticus associated with white spot disease of Penaeus monodon.  

PubMed

In February 2000, white spot disease outbreaks occurred among cultured Penaeus monodon in extensive shrimp farms on the southwest coast of India. Bacteria were isolated from infected shrimp that showed reddish body coloration and white spots in the cuticle. The isolates were screened on thiosulfate citrate bile salt sucrose (TCBS) agar plates for the selection of Vibrio species. The primary isolate (QS7) was characterized as V. alginolyticus based on morphological, biochemical and physiological characteristics. Antibiotic sensitivity tests of QS7 indicated that the isolate was highly sensitive to chloramphenicol, ciprofloxacin, nalidixic acid and streptomycin. Pathogenicity tests confirmed that the isolate was virulent for P. monodon. Based on the lethal dose (LD50) value (5 x 10(6) cfu per shrimp), it was inferred that shrimp weakened by white spot syndrome virus would succumb to secondary infection by QS7. PMID:14735933

Selvin, Joseph; Lipton, A P

2003-12-01

381

The Virulence Activator AphA Links Quorum Sensing to Pathogenesis and Physiology in Vibrio cholerae by Repressing the Expression of a Penicillin Amidase Gene on the Small Chromosome  

Microsoft Academic Search

Activation of the tcpPH promoter on the Vibrio pathogenicity island by AphA and AphB initiates the Vibrio cholerae virulence cascade and is regulated by quorum sensing through the repressive action of HapR on aphA expression. To further understand how the chromosomally encoded AphA protein activates tcpPH expression, site-directed mutagenesis was used to identify the base pairs critical for AphA binding

Gabriela Kovacikova; Wei Lin; Karen Skorupski

2003-01-01

382

Differentiation of Environmental and Clinical Isolates of Vibrio mimicus from Vibrio cholerae by Multilocus Enzyme Electrophoresis  

Microsoft Academic Search

In this study, we demonstrated that analyzed strains of Vibrio mimicus and Vibrio cholerae could be separated in two groups by using multilocus enzyme electrophoresis (MEE) data from 14 loci. We also showed that the combination of four enzymatic loci enables us to differentiate these two species. Our results showed that the ribosomal intergenic spacer regions PCR-mediated identification system failed,

VERONICA V. VIEIRA; LUIZ FERNANDO M. TEIXEIRA; ANA CAROLINA P. VICENTE; HOOMAN MOMEN; C. A. Salles

2001-01-01

383

Vibrio chromosomes share common history  

PubMed Central

Background While most gamma proteobacteria have a single circular chromosome, Vibrionales have two circular chromosomes. Horizontal gene transfer is common among Vibrios, and in light of this genetic mobility, it is an open question to what extent the two chromosomes themselves share a common history since their formation. Results Single copy genes from each chromosome (142 genes from chromosome I and 42 genes from chromosome II) were identified from 19 sequenced Vibrionales genomes and their phylogenetic comparison suggests consistent phylogenies for each chromosome. Additionally, study of the gene organization and phylogeny of the respective origins of replication confirmed the shared history. Conclusions Thus, while elements within the chromosomes may have experienced significant genetic mobility, the backbones share a common history. This allows conclusions based on multilocus sequence analysis (MLSA) for one chromosome to be applied equally to both chromosomes.

2010-01-01

384

Lessons from cholera & Vibrio cholerae  

PubMed Central

Cholera is an acute form of diarrhoeal disease that plagued human civilization over the centuries. The sudden and explosive onset of the disease in the form of an outbreak or epidemic, coupled with high mortality and morbidity rates, had a tragic impact on the personal as well as social life of people living in the affected areas. The enormity of human sufferings led clinicians and scientists to carry out extensive research on cholera and Vibrio cholerae (the causative bacterium of the disease) leading to major discoveries that opened up novel areas of research or new disciplines in biomedical sciences. An attempt is made here to summarize some of these breakthroughs and outline their significance in broader perspectives. Finally, the possible impact of the global socio-political scenario on the spread of cholera epidemics (pandemicity of cholera) is briefly discussed.

Ghose, Asoke C.

2011-01-01

385

Construction of a Vibrio splendidus mutant lacking the metalloprotease gene vsm by use of a novel counterselectable suicide vector.  

PubMed

Vibrio splendidus is a dominant culturable Vibrio in seawater, and strains related to this species are also associated with mortality in a variety of marine animals. The determinants encoding the pathogenic properties of these strains are still poorly understood; however, the recent sequencing of the genome of V. splendidus LGP32, an oyster pathogen, provides an opportunity to decipher the basis of the virulence properties by disruption of candidate genes. We developed a novel suicide vector based on the pir-dependent R6K replicative origin, which potentially can be transferred by RP4-based conjugation to any Vibrio strain and which also carries the plasmid F toxin ccdB gene under control of the PBAD promoter. We demonstrated that this genetic system allows efficient counterselection of integrated plasmids in the presence of arabinose in both V. splendidus and Vibrio cholerae and thus permits efficient markerless allelic replacement in these species. We used this technique to construct several mutants of V. splendidus LGP32, including a derivative with a secreted metalloprotease gene, vsm, deleted. We found that this gene is essential for LGP32 extracellular product toxicity when the extracellular products are injected into oysters but is not necessary for virulence of bacteria in the oyster infection model when bacteria are injected. PMID:17122399

Le Roux, Frédérique; Binesse, Johan; Saulnier, Denis; Mazel, Didier

2007-02-01

386

Isolation, identification, and characterization of Vibrio cholerae from the Danube River in Slovakia.  

PubMed

The occurrence of Vibrio cholerae, an important aquatic pathogen, was assessed in the surface water of the Danube River near Bratislava. The isolates were distinguished by biochemical tests and grouped by ARDRA to three clusters corresponding to three species (V. cholerae, Vibrio metschnikovii, and Aeromonas spp.). The identification of V. cholerae was confirmed by multiplex PCR using primer pairs targeted to ompW gene (membrane protein), ctxA gene (toxicity gene), and toxR gene (regulatory gene). None from the isolated V. cholerae from surface water contained ctxA gene; seven of them possessed toxR gene. Serotyping of V. cholerae isolates with polyvalent O antiserum and O/139 antiserum was negative. All isolates of V. cholerae were susceptible to chloramphenicol, rifampicin, tetracycline, variable to ampicillin, and resistant to kanamycin and streptomycin. PMID:22457232

Seman, Milan; Prokšová, Miloslava; Rosinský, Jozef; Ferianc, Peter

2012-05-01

387

Arp2/3-independent assembly of actin by Vibrio type III effector VopL  

PubMed Central

Microbial pathogens use a variety of mechanisms to disrupt the actin cytoskeleton during infection. Vibrio parahaemolyticus (V. para) is a Gram-negative bacterium that causes gastroenteritis, and new pandemic strains are emerging throughout the world. Analysis of the V. para genome revealed a type III secretion system effector, VopL, encoding three Wiskott–Aldrich homology 2 domains that are interspersed with three proline-rich motifs. Infection of HeLa cells with V. para induces the formation of long actin fibers in a VopL-dependent manner. Transfection of VopL promotes the assembly of actin stress fibers. In vitro, recombinant VopL potently induces assembly of actin filaments that grow at their barbed ends, independent of eukaryotic factors. Vibrio VopL is predicted to be a bacterial virulence factor that disrupts actin homeostasis during an enteric infection of the host.

Liverman, Amy D. B.; Cheng, Hui-Chun; Trosky, Jennifer E.; Leung, Daisy W.; Yarbrough, Melanie L.; Burdette, Dara L.; Rosen, Michael K.; Orth, Kim

2007-01-01

388

Identification and Characterization of RbmA, a Novel Protein Required for the Development of Rugose Colony Morphology and Biofilm Structure in Vibrio cholerae  

Microsoft Academic Search

Phase variation between smooth and rugose colony variants of Vibrio cholerae is predicted to be important for the pathogen's survival in its natural aquatic ecosystems. The rugose variant forms corrugated colonies, exhibits increased levels of resistance to osmotic, acid, and oxidative stresses, and has an enhanced capacity to form biofilms. Many of these phenotypes are mediated in part by increased

Jiunn C. N. Fong; Kevin Karplus; Gary K. Schoolnik; Fitnat H. Yildiz

2006-01-01

389

Effect of fish skin mucus on the soluble proteome of Vibrio salmonicida analysed by 2-D gel electrophoresis and tandem mass spectrometry  

Microsoft Academic Search

Vibrio salmonicida is the causative agent of cold-water vibriosis in farmed marine fish species. Adherence of pathogenic bacteria to mucosal surfaces is considered to be the first steps in the infective processes, and proteins involved are regarded as virulence factors. The global protein expression profile of V. salmonicida, grown with and without the presence of fish skin mucus in the

Inger Lin Uttakleiv Ræder; Steinar M. Paulsen; Arne O. Smalås; Nils Peder Willassen

2007-01-01

390

Control of luminous Vibrio species in penaeid aquaculture ponds  

Microsoft Academic Search

A crisis has arisen in the prawn industry in many regions with the onset of disease, with Vibrio spp. being important major causal factors. The value of adding selected strains of Bacillus as probiotic bacteria to control the Vibrio is shown by comparing farms in Indonesia using the same water sources, which contained luminous Vibrio strains. The farms that did

D. J. W Moriarty

1998-01-01

391

Evaluation of bactericidal activity of weakly acidic electrolyzed water (WAEW) against Vibrio vulnificus and Vibrio parahaemolyticus.  

PubMed

Vibrio parahaemolyticus and Vibriovulnificus cause severe foodborne illness in humans; thus, to reduce outbreaks of disease, it is clearly important to reduce food contamination by these pathogens. Although electrolyzed oxidizing (EO) water has been reported to exhibit strong bactericidal activities against many pathogens, it has never been tested against V. vulnificus and V. parahaemolyticus. The purpose of this study was to evaluate the bactericidal activity of weakly acidic electrolyzed water (WAEW), a type of EO water, against V. vulnificus and V. parahaemolyticus. Cell suspensions and cell cultures of both pathogens were treated for 30s with sodium hypochlorite solution containing 35mg/L available chlorine concentration (ACC) or WAEW containing 35mg/L ACC. After an initial inoculum of 5.7logCFU/mL, the number of viable V. vulnificus cells was reduced by 2.2 logs after treatment for 60s with sodium hypochlorite solution containing 35mg/L ACC, while no cells survived treatment with WAEW for 30s. Similar results were obtained for V. parahaemolyticus. Under open storage conditions, WAEW maintained bactericidal activities against cell suspensions of both strains after 5weeks but disappeared against cell cultures of the two strains after 5weeks. Under closed storage conditions, however, WAEW maintained bactericidal activities against both cell suspensions and cell cultures of each strain after 5weeks. No cells were detected in the cell suspensions and cultures when the ACC of WAEW was more than 20mg/L and treatment time was greater than 15s. Bactericidal activity of WAEW against V. vulnificus cell culture was reduced when the ACC of WAEW was less than 15mg/L but was maintained in the V. vulnificus cell suspension when the ACC of WAEW was 0.5mg/L. Thus, the bactericidal activity of WAEW was primarily affected by ACC rather than treatment time. Similar results were obtained for V. parahaemolyticus, indicating that WAEW kills these microorganisms more quickly than a chemical product such as sodium hypochlorite (NaClO), even at equivalent ACCs. PMID:20004034

Quan, Yaru; Choi, Kyoo-Duck; Chung, Donghwa; Shin, Il-Shik

2010-01-01

392

Identification of vibrio cholerae and vibrio mimicus by multilocus sequence analysis (MLSA).  

PubMed

Vibrio cholerae and Vibrio mimicus have similar phenotypes and genomes making rapid differentiation of these two species difficult. The first standard multilocus sequence analysis (MLSA) scheme for the identification of these species is described. A collection of 45 representative isolates from different geographical regions and hosts was examined using segments of the housekeeping genes pyrH, recA and rpoA. Overall, the closest phylogenetic neighbours of these species were Vibrio furnissii and Vibrio fluvialis. V. cholerae and V. mimicus formed separate species clusters on the basis of each gene, suggesting that these genes are useful as identification markers. These species clusters arose by the accumulation of point mutations. The pyrH gene showed the highest resolution for differentiating V. cholerae and V. mimicus. The maximum interspecies pyrH gene sequence similarity was 91 %. Clearly, V. mimicus strains were more heterogeneous than V. cholerae strains at the three loci. It is suggested that vibrio species may be defined on the basis of MLSA data. A vibrio species was defined as a group of strains forming a monophyletic group on the basis of these loci and with an intraspecific sequence similarity of at least 95 %. V. cholerae and V. mimicus isolates can be readily identified through the open database resource 'The Taxonomy of Vibrios' (http://www.taxvibrio.lncc.br/). PMID:18319466

Thompson, Cristiane C; Thompson, Fabiano L; Vicente, Ana Carolina P

2008-03-01

393

Subversion of inflammasome activation and pyroptosis by pathogenic bacteria  

PubMed Central

Activation of the inflammasome occurs in response to a notably high number of pathogenic microbes and is a broad innate immune response that effectively contributes to restriction of pathogen replication and generation of adaptive immunity. Activation of these platforms leads to caspase-1- and/or caspase-11-dependent secretion of proteins, including cytokines, and induction of a specific form of cell death called pyroptosis, which directly or indirectly contribute for restriction of pathogen replication. Not surprisingly, bona fide intracellular pathogens developed strategies for manipulation of cell death to guarantee intracellular replication. In this sense, the remarkable advances in the knowledge of the inflammasome field have been accompanied by several reports characterizing the inhibition of this platform by several pathogenic bacteria. Herein, we review some processes used by pathogenic bacteria, including Yersinia spp., Pseudomonas aeruginosa, Vibrio parahaemolyticus, Chlamydia trachomatis, Francisella tularensis, Shigella flexneri, Legionella pneumophila, and Coxiella burnetii to evade the activation of the inflammasome and the induction of pyroptosis.

Cunha, Larissa D.; Zamboni, Dario S.

2013-01-01

394

Antibacterial activity of citrus fruit juices against Vibrio species.  

PubMed

Lemon, lime and sudachi juices were tested for antibacterial activity against seven strains of Vibrio species. All juices were effective in inhibiting the growth of the Vibrio strains. Citric acid, the major organic acid in these juices, was found to be responsible for inhibiting the growth of Vibrio parahaemolyticus. Sauce prepared from sudachi juice showed a strong bactericidal activity against Vibrio parahaemolyticus, whereas the sauce adjusted to higher pH values had no bacterial activity. Diluted sudachi juice or citric acid solution also had antibacterial activity independently. These results suggest that citrus fruit juices are effective in preventing infection with Vibrio species. PMID:16802698

Tomotake, Hiroyuki; Koga, Tetsuro; Yamato, Masayuki; Kassu, Afework; Ota, Fusao

2006-04-01

395

Sensitive and rapid detection of cholera toxin-producing Vibrio cholerae using a loop-mediated isothermal amplification  

Microsoft Academic Search

BACKGROUND: Vibrio cholerae is widely acknowledged as one of the most important waterborne pathogen causing gastrointestinal disorders. Cholera toxin (CT) is a major virulence determinant of V. cholerae. Detection of CT-producing V. cholerae using conventional culture-, biochemical- and immunological-based assays is time-consuming and laborious, requiring more than three days. Thus, we developed a novel and highly specific loop-mediated isothermal amplification

Wataru Yamazaki; Kazuko Seto; Masumi Taguchi; Masanori Ishibashi; Kiyoshi Inoue

2008-01-01

396

The Vibrio fischeri sapABCDF locus is required for normal growth, both in culture and in symbiosis  

Microsoft Academic Search

Inactivation of thesapABCDF genes results in a loss of virulence in several bacterial pathogens of animals and plants. The role of this locus in the\\u000a growth physiology ofVibrio fischeri, and in the symbiotic colonization of the squidEuprymna scolopes was investigated. In rich medium, aV. fischeri sapA insertion mutant grew at only 85% the rate of its wild-type parent. While a

Claudia Lupp; Robert E. W. Hancock; Edward G. Ruby

2002-01-01

397

Characterization of htrB and msbB Mutants of the Light Organ Symbiont Vibrio fischeri  

Microsoft Academic Search

Bacterial lipid A is an important mediator of bacterium-host interactions,