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identification of lacto-n-biose i phosphorylase from vibrio vulnificus cmcp6.a beta-1,3-galactosyl-n-acetylhexosamine phosphorylase (galglynacp) homolog gene was cloned from vibrio vulnificus cmcp6. in synthetic reactions, the recombinant enzyme acted only with glcnac and galnac as acceptors in the presence of alpha-d-galactose-1-phosphate as a donor to form lacto-n-biose i (lnb) (galbeta1 --> 3glcnac) and galacto-n-biose (gnb) (galbeta1 --> 3galnac), respectively. glcnac was a much better acceptor than galnac. the enzyme also phosphorolysed lnb faster than it phosphorol ...200818723650
vibrio vulnificus rpos expression is repressed by direct binding of camp-camp receptor protein complex to its two promoter regions.vibrio vulnificus, a septicemia-causing pathogenic bacterium, acquires resistance against various stresses and expresses virulence factors via an rpos gene product. in this study, we investigated the transcriptional characteristics of this global regulator. two distinct transcriptional initiation sites for the rpos gene, the proximal promoter (p(p)) and the distal promoter (p(d)), were defined by primer extension experiments. various rpos::luxab transcriptional fusions indicated that p(d) is a m ...200818713737
differentiating between isolates of vibrio vulnificus with monoclonal antibodies.monoclonal antibodies (mabs) against vibrio vulnificus (isolate i, vvc and isolate ii, vvb) were raised using heat-killed and heat-killed plus sds-mercaptoethanol treated forms of vvc and vvb for immunizing swiss mice. twenty three hybridomas producing mabs against v. vulnificus were selected and divided into five groups according to their specificities to different v. vulnificus isolates and apparent protein antigens which ranged from approximately 3-50 kda. four groups were specific to v. vuln ...200818706941
detection of quorum-sensing-related molecules in vibrio scophthalmi.cell-to-cell communication (also referred to as quorum sensing) based on n-acyl-homoserine lactones (ahls) is a widespread response to environmental change in gram-negative bacteria. ahls seem to be highly variable, both in terms of the acyl chain length and in the chemical structure of the radicals. another quorum sensing pathway, the autoinducer-2-based system, is present both in gram-positive and gram-negative bacteria. in this study the presence of signal molecules belonging to both quorum s ...200818700048
multiplex pcr assay for detection and simultaneous differentiation of genotypes of vibrio vulnificus biotype 1.a multiplex pcr assay was developed that provides a rapid method of species identification with simultaneous genotype differentiation of vibrio vulnificus biotype 1. this technique has been employed for the confirmation and typing of unknown isolates from both clinical and environmental sources utilizing a single reaction followed by gel electrophoresis.200818687035
effectiveness of icing as a postharvest treatment for control of vibrio vulnificus and vibrio parahaemolyticus in the eastern oyster (crassostrea virginica).the focus of this research was to investigate the efficacy of icing as a postharvest treatment for reduction of the levels of vibrio vulnificus and vibrio parahaemolyticus in commercial quantities of shellstock oysters. the experiments were conducted in june and august of 2006 and consisted of the following treatments: (i) on-board icing immediately after harvest; (ii) dockside icing approximately 1 to 2 h prior to shipment; and (iii) no icing (control). changes in the levels of pathogenic vibri ...200818680950
multi-site analysis reveals widespread antibiotic resistance in the marine pathogen vibrio vulnificus.vibrio vulnificus is a serious opportunistic human pathogen commonly found in subtropical coastal waters, and is the leading cause of seafood-borne mortality in the usa. this taxon does not sustain prolonged presence in clinical or agricultural settings, where it would undergo human-induced selection for antibiotic resistance. therefore, few studies have verified the effectiveness of commonly prescribed antibiotics in v. vulnificus treatment. here we screened 151 coastal isolates and 10 primary ...200918642041
proteomic identification and characterization of vibrio vulnificus proteins induced upon exposure to int-407 intestinal epithelial cells.proteomic analysis led to identification of the proteins of vibrio vulnificus that were induced upon exposure to int-407 cells, and 7 of which belong to the functional categories such as amino acid transport/metabolism, nucleotide transport/metabolism, posttranslational modification/protein turnover/chaperones, and translation. among the genes encoding the host-induced proteins, disruption of purh, trpd, tsaa, and groel2 resulted in reduced cytotoxicity. the purh, trpd, and tsaa mutants showed i ...200818633300
a case of vibrio vulnificus keratitis. 200818626738
structure-function relationships in escherichia coli adenylate cyclase.class i adenylate cyclases are found in gamma- and delta-proteobacteria. they play central roles in processes such as catabolite repression in escherichia coli or development of full virulence in pathogens such as yersinia enterocolitica and vibrio vulnificus. the catalytic domain (residues 2-446) of the adenylate cyclase of e. coli was overexpressed and purified. it displayed a v(max) of 665 nmol of camp x mg(-1) x min(-1) and a k(m) of 270 microm. titration of the metal cofactor mg(2+) against ...200818620542
comparison of conventional, nested, and real-time pcr assays for rapid and accurate detection of vibrio vulnificus.we conducted a prospective study to target toxr in the blood of patients with skin and soft tissue infections who were admitted to four tertiary hospitals to assess the clinical usefulness of real-time quantitative pcr (q-pcr) as a diagnostic technique. we performed conventional pcr (c-pcr), nested pcr (n-pcr), and q-pcr assays and compared the results to those obtained using the "gold standard" of microbiological culture. the lower detection limit for the q-pcr assay was 5 x 10(0) copies/microl ...200818614647
two forms of vibrio vulnificus metalloprotease vvpe are secreted via the type ii general secretion system.vibrio vulnificus has been known to secrete one form of metalloprotease vvpe (45 kda) that is cleaved to 34 kda-vvpe and 11 kda-c-terminal propeptide via extracellular autoproteolysis. however, we found that extracellular secretion of both the 34 and 45 kda forms of vvpe began in the early growth phase; moreover, 34 kda-vvpe existed as the major form in v. vulnificus cell lysates and culture supernatants. in addition, extracellular secretion of both 34 and 45 kda-vvpe was blocked by mutation of ...200818604505
sensitive and rapid identification of vibrio vulnificus by loop-mediated isothermal amplification.vibrio vulnificus is a serious bacterial pathogen for humans and aquatic animals. we developed a rapid, sensitive and specific identification method for v. vulnificus using loop-mediated isothermal amplification (lamp) technique. a set of primers, composed of two outer primers and two inner primers, was designed based on the cytolysin gene sequence of v. vulnificus. the lamp reaction was processed in a heat block at 65 degrees c for 60 min. the amplification products were detected by visual insp ...200918585020
a consensus sequence for binding of smcr, a vibrio vulnificus luxr homologue, and genome-wide identification of the smcr regulon.quorum sensing has been implicated as an important global regulatory system controlling the expression of numerous virulence factors in bacterial pathogens. in the present study, dna targets of smcr, a vibrio vulnificus luxr homologue, were selected from a random pool of dna fragments by using a cycle selection procedure consisting of in vitro dna-smcr interaction, purification of smcr-dna complexes, and pcr amplification of smcr-bound dna. the amplified dna fragments were cloned and analyzed se ...200818579523
global gene expression as a function of the iron status of the bacterial cell: influence of differentially expressed genes in the virulence of the human pathogen vibrio vulnificus.vibrio vulnificus multiplies rapidly in host tissues under iron-overloaded conditions. to understand the effects of iron in the physiology of this pathogen, we performed a genome-wide transcriptional analysis of v. vulnificus growing at three different iron concentrations, i.e., iron-limiting [trypticase soy broth with 1.5% nacl (tsbs) plus ethylenediamine-di-(o-hydroxyphenylacetic) acid (edda)], low-iron (1 microg fe/ml; tsbs), and iron-rich (38 microg fe/ml; tsbs plus ferric ammonium citrate) ...200818573903
vibrio vulnificus infection presenting with compartment syndrome of bilateral calves. 200818572348
vibrio vulnificus-induced death of jurkat t-cells requires activation of p38 mitogen-activated protein kinase by nadph oxidase-derived reactive oxygen species.vibrio vulnificus, a pathogenic bacterium causing primary septicemia, exhibited cytotoxicity towards jurkat cells of t-lymphocytes through intracellular reactive oxygen species (ros) production. pretreatment of jurkat t-cells with diphenyleneiodonium chloride (dpi) abolished v. vulnificus-induced ros generation and bacterial ability to cause cell death. jurkat t-cells expressing dominant-negative protein of rac subunit of nadph oxidase (nox) did not show increased ros production and cell death b ...200818571150
evaluation of a loop-mediated isothermal amplification assay for detecting vibrio vulnificus in raw oysters.human consumption of raw or undercooked seafood, particularly oysters, may lead to severe infections due to the presence of vibrio vulnificus. in this study, a sensitive and specific loop-mediated isothermal amplification (lamp) assay was developed to detect this pathogen in raw oysters. two outer and two inner primers were designed to specifically recognize the v. vulnificus cytolysin/hemolysin gene (vvha), and the reaction could be completed in 1 hour at 63 degrees c. direct visual observation ...200818564911
the ecology of vibrio vulnificus, vibrio cholerae, and vibrio parahaemolyticus in north carolina estuaries.while numerous studies have characterized the distribution and/or ecology of various pathogenic vibrio spp., here we have simultaneously examined several estuarine sites for vibrio vulnificus, v. cholerae, and v. parahaemolyticus. for a one year period, waters and sediment were monitored for the presence of these three pathogens at six different sites on the east coast of north carolina in the united states. all three pathogens, identified using colony hybridization and pcr methods, occurred in ...200818545963
on the origin and function of an insertion element vpai-1 specific to post-1995 pandemic vibrio parahaemolyticus strains.since 1995, new virulent strains of vibrio parahaemolyticus have emerged and spread throughout the world. these "pandemic" strains have four strain specific genomic islands (gis), which are considered to be potential factors of the pandemicity. we investigated the origin and function of 24 genes in the so-called vpai-1, one of the four gis, by searching homologs in various species in bacteria and archaea. of these 24 genes, two are found only in vibrio vulnificus cmcp6 and shewanella sp. mr-7. t ...200818506094
necrotising fasciitis caused by vibrio vulnificus in a man with cirrhosis. 200818501855
[septic shock after exposure to vibrio vulnificus].vibrio vulnificus (vv) is a rare cause of infections in denmark. in this case, a patient suffering from chronic diseases developed wound infection, severe sepsis and despite adequate treatment, septic shock after exposure to seawater and fish. the fact that the condition was caused by vv was not identified until the results of the blood cultures. vv is the cause of primary sepsis, wound infections and gastroenteritis. the infection is treated with fluid resuscitation, broad-spectrum antibiotics ...200818492457
cyclic-di-gmp regulates extracellular polysaccharide production, biofilm formation, and rugose colony development by vibrio vulnificus.vibrio vulnificus is a human and animal pathogen that carries the highest death rate of any food-borne disease agent. it colonizes shellfish and forms biofilms on the surfaces of plankton, algae, fish, and eels. greater understanding of biofilm formation by the organism could provide insight into approaches to decrease its load in filter feeders and on biotic surfaces and control the occurrence of invasive disease. the capsular polysaccharide (cps), although essential for virulence, is not requi ...200818487410
inducible resistance of fish bacterial pathogens to the antimicrobial peptide cecropin b.cecropin b is a cationic antimicrobial peptide originally isolated from the diapausing pupae of the giant silk moth, hylphora cecropia. cecropin b elicits its antimicrobial effects through disruption of the anionic cell membranes of gram-negative bacteria. previous work by our laboratory demonstrated that a constitutively expressed cecropin b transgene conferred enhanced resistance to bacterial infection in medaka. the development of antibiotic resistance by pathogenic bacteria is a growing prob ...200818474580
achilles tendinitis in vibrio vulnificus sepsis: a rare presentation.vibrio vulnificus infection commonly presents as sepsis or wound infection. a case of alcoholic liver cirrhosis developed acute left heel pain and swelling 2 days after ingesting raw fish. two sets of blood cultures indicated v. vulnificus. hla-b27 was negative. ultrasonography revealed left achilles tendinitis. fluid aspirated from the left retrocalcaneal bursa was sterile. the patient recovered completely 3 weeks after treatment. to our knowledge, v. vulnificus septicaemia with achilles tendin ...200818465460
lower extremity infections by vibrio vulnificus.vibrio vulnificus is a gram-negative pathogen which is found in seawater and shellfish during warm months and can cause local infections in healthy individuals or septicemia in patients with chronic liver disease. materials-methods: clinical and laboratory records of four complicated cases are presented, with a 4.2 mean year follow-up.200818457099
the extracellular metalloprotease of vibrio tubiashii is a major virulence factor for pacific oyster (crassostrea gigas) larvae.vibrio tubiashii is a recently reemerging pathogen of larval bivalve mollusks, causing both toxigenic and invasive disease. marine vibrio spp. produce an array of extracellular products as potential pathogenicity factors. culture supernatants of v. tubiashii have been shown to be toxic to oyster larvae and were reported to contain a metalloprotease and a cytolysin/hemolysin. however, the structural genes responsible for these proteins have yet to be identified, and it is uncertain which extracel ...200818456850
nonfoodborne vibrio infections: an important cause of morbidity and mortality in the united states, 1997-2006.infections due to vibrio species cause an estimated 8000 illnesses annually, often through consumption of undercooked seafood. like foodborne vibrio infections, nonfoodborne vibrio infections (nfvi) also result in serious illness, but awareness of these infections is limited.200818444811
vibrio vulnificus septicemia after handling tilapia species fish: a canadian case report and review.vibrio vulnificus can cause a necrotizing soft tissue infection or primary septicemia; these infections are collectively known as vibriosis. this bacterium is commonly found within molluscan shellfish. primary septicemia is often fatal, principally affecting persons with chronic liver disease.200618418489
variable repeat regions in the genome of vibrio vulnificus and polymorphism in one of the loci in strains isolated from oysters.vibrio vulnificus an estuarine bacterium is associated with severe wound infections and fatal septicemia related to consumption of raw shellfish. in this study we screened the two whole genome sequences available for v. vulnificus in genbank for the presence of variable number of tandem repeat (vntr) regions. five potential vntr loci with unit repeat size ranging from 6-7 nucleotides were identified for v. vulnificus genome. one of the loci designated vv1 was selected to detect the repeat number ...200818374439
bacteria flora and heavy metals in cultivated oysters crassostrea iredalei of setiu wetland, east coast peninsular malaysia.slipper oyster crassostrea iredalei is a species of good demand for its sweet flavor and white coloured flesh. the filter feeding nature predisposes oysters to accumulation of pathogenic and heavy metals in waters impacted by sewage pollutions and may thus render the oysters unfit for human consumption. a study was undertaken to investigate the presence of bacteria flora and heavy metal concentrations in cultivated oysters crassostrea iredalei at setiu wetland, terengganu, the only source of cul ...200818369732
phylogenetic analysis of the incidence of lux gene horizontal transfer in vibrionaceae.horizontal gene transfer (hgt) is thought to occur frequently in bacteria in nature and to play an important role in bacterial evolution, contributing to the formation of new species. to gain insight into the frequency of hgt in vibrionaceae and its possible impact on speciation, we assessed the incidence of interspecies transfer of the lux genes (luxcdabeg), which encode proteins involved in luminescence, a distinctive phenotype. three hundred three luminous strains, most of which were recently ...200818359809
diagnosing and treating deadly vibrio vulnificus infection. 200818358354
occurrence, seasonality and genetic diversity of vibrio vulnificus in coastal seaweeds and water along the kii channel, japan.vibrio vulnificus is a ubiquitous toxigenic bacterium found in a coastal environment but little is known about its occurrence and seasonality among seaweeds, which are widely consumed as seafood in japan. therefore, we have observed the bacterium's abundance in seawater and seaweed samples from three areas of the kii channel, japan, during june 2003 to may 2004. a total of 192 samples were collected: 24 from each source in summer, autumn, winter and spring. the samples were selectively cultivate ...200818355295
epidemiological survey of vibrio vulnificus infection in japan between 1999 and 2003.the frequency of vibrio vulnificus infection is very rare and there are many questions regarding its epidemiology in japan. to investigate the clinical course and epidemiology of v. vulnificus infection in japan, we performed a retrospective questionnaire survey in which 1693 hospitals from all over japan were surveyed, including advanced life saving emergency centers and dermatology institutions. of the 1693 hospitals, we received answers from 1045. ninety-four cases were confirmed as v. vulnif ...200818346255
evidence that aphb, essential for the virulence of vibrio vulnificus, is a global regulator.the vibrio vulnificus aphb mutant was significantly less virulent than the wild type and was impaired in motility and adherence to host cells. microarray analysis revealed that aphb of v. vulnificus (aphb(vv)) influences the expression of over 10% of the v. vulnificus genome. the combined results indicated that aphb(vv) is a global regulator contributing to the pathogenesis of v. vulnificus.200818344367
characterization of the low-salinity stress in vibrio vulnificus.vibrio vulnificus is a marine pathogenic bacterium commonly found in seawater or seafood. this organism encounters low-salinity stress in its natural environment and during food processing. this study was designed to investigate the response of v. vulnificus yj03 to lethal low salinity (0.04% nacl) and its adaptation to sublethal salinity (0.12% nacl with 20 amino acids added). a short period in the nonculturable state was induced by lethal low-salinity stress followed by cell death after 30 min ...200818326198
evolution of exceptionally large genes in prokaryotes.analysis of bacterial genomic sequences revealed an average bacterial gene size of approximately 1 kb. however, genes with a size >10 kb were also noted. this study investigates the prevalence, possible function, and origin of exceptionally large-size genes (elsgs; >10 kb) in prokaryotes. forty-two elsgs (0.03%) were identified after searching more than 170,000 genes in 46 bacterial and 11 archaeal species. these elsgs were found in diverse species including both archaeal and eubacterial kingdom ...200818322635
variation of extracellular proteases produced by vibrio vulnificus clinical isolates: genetic diversity of the metalloprotease gene (vvp), and serine protease secretion by vvp-negative strains.vibrio vulnificus is a causative agent of septicemia or wound infection in human and eel; however, the genetic variation between human and eel isolates has been reported. in the present study, the difference in the vvp gene encoding a tissue-damaging metalloprotease was investigated. the gene of strain e86 from a diseased eel (type b vvp) was 95.2% identical with that of strain l-180 from human blood (type a vvp). pcr using oligonucleotide primers designed to differentiate two types of the gene ...200818262748
new approach for discrimination of vibrio vulnificus by real-time pcr before and after gamma-irradiation.the effects of gamma-irradiation on the destruction of vibrio vulnificus by real-time pcr were studied. gamma-irradiation was found to result in extensive reduction in the molecular size of dna. irradiation of viable cells (1 x 10(6) cfu/ml) at 1.08 kgy resulted in 100% destruction determined by plate counts, with most of the dna from the irradiated cells having a bp-length of less than 1000. the use of a pair of primers to amplify a 1000-bp sequence of dna from cells exposed to 1.08 kgy failed ...200818262296
autopsy cases of fulminant-type bacterial infection with necrotizing fasciitis: group a (beta) hemolytic streptococcus pyogenes versus vibrio vulnificus infection.two autopsy cases of fulminant-type infection associated with necrotizing fasciitis were analyzed clinicopathologically. both cases involved 57-year-old alcohol abusers. the former was a woman with group a (beta) hemolytic streptococcus pyogenes infection, and the latter was a man with vibrio vulnificus infection. the sudden onset of shock with high fever resulted in sepsis, decreased clotting, and hepatorenal symptoms, followed by death within a few days. post-mortem examination showed widespre ...200818251785
vibrio vulnificus rtxe is important for virulence, and its expression is induced by exposure to host cells.numerous secreted virulence factors have been proposed to account for the fulminating and destructive nature of vibrio vulnificus infections. a mutant of v. vulnificus that exhibited less cytotoxicity to int-407 human intestinal epithelial cells was screened from a library of mutants constructed by random transposon mutagenesis. a transposon-tagging method was used to identify and clone an open reading frame encoding an rtx toxin secretion atp binding protein, rtxe, from v. vulnificus. the deduc ...200818250174
real-time pcr assays for quantification and differentiation of vibrio vulnificus strains in oysters and water.vibrio vulnificus is an autochthonous estuarine bacterium and a pathogen that is frequently transmitted via raw shellfish. septicemia can occur within 24 h; however, isolation and confirmation from water and oysters require days. real-time pcr assays were developed to detect and differentiate two 16s rrna variants, types a and b, which were previously associated with environmental sources and clinical fatalities, respectively. both assays could detect 10(2) to 10(3) v. vulnificus total cells in ...200818245234
nonribosomal peptide synthase is responsible for the biosynthesis of siderophore in vibrio vulnificus mo6-24/o.vibrio vulnificus produces siderophores, lowmolecular- weight iron-chelating compounds, to obtain iron under conditions of iron deprivation. to identify genes associated with the biosynthesis of siderophore in v. vulnificus mo6-24/ o, we screened clones with mini-tn5 random insertions for those showing decreased production of siderophore. among 6,000 clones screened, nine such clones were selected. these clones contain the transposon inserted in vv2_0830 (genbank accession number) that is a homo ...200818239413
gene expression of cold shock and other stress-related genes in vibrio vulnificus grown in pure culture under shellstock temperature control conditions.shellstock refrigeration after harvesting is recommended to prevent further increases in vibrio vulnificus numbers in oysters, but it could potentially induce a cold shock response in this bacterium. v. vulnificus was incubated at 35, 25, 20, and 15 degrees c and then subjected to 7.2 and 4 degrees c for 1 week. a cold-adaptation response that enhanced cell culturability was observed when cells were incubated at 15 degrees c prior to cold shock at 7.2 degrees c. in vitro cold shock gene expressi ...200818236677
vibrio vulnificus biotype 2 serovar e gne but not gale is essential for lipopolysaccharide biosynthesis and virulence.this work aimed to establish the role of gne (encoding udp-galnac 4-epimerase activity) and gale (encoding udp-gal-4-epimerase activity) in the biosynthesis of surface polysaccharides, as well as in the virulence for eels and humans of the zoonotic serovar of vibrio vulnificus biotype 2, serovar e. dna sequence data revealed that gne and gale are quite homologous within this species (> or =90% homology). mutation in gne of strain cect4999 increased the surface hydrophobicity, produced deep alter ...200818227162
further characterization of vibrio vulnificus rugose variants and identification of a capsular and rugose exopolysaccharide gene cluster.capsular polysaccharide (cps) is a major virulence factor in vibrio vulnificus, and encapsulated strains have an opaque, smooth (ops) colony morphology, while nonencapsulated strains have a translucent, smooth (trs) colony morphology. previously, we showed that ops and trs parental strains can yield a third colony type, rugose (r), and that the resulting strains, with the opr and trr phenotypes, respectively, form copious biofilms. here we show that while opr and trr strains both produce three-d ...200818212074
[a protocol for diagnosis and treatment of vibrio vulnificus sepsis].vibrio vulnificus sepsis is one of the most fatal disease with a high mortality which exceeds 50%. but at present there is no evidence-based guidelines for diagnosis and therapy of vibrio vulnificus sepsis because of its dispersion in occurrence and low incidence.200818199391
[sepsis and cellulitis by vibrio vulnificus in cirrhotic patient]. 200718198957
conditions for a 5-log reduction of vibrio vulnificus in oysters through high hydrostatic pressure treatment.vibrio vulnificus is frequently associated with oysters, and since oysters are typically consumed raw on a half shell, they can pose a threat to public health due to ingestion of this pathogenic marine microorganism. oysters should be processed to reduce the number of this pathogen. high pressure processing is gaining more and more acceptance among oyster processors due to its ability to shuck oysters while keeping the fresh-like characteristics of oysters. nine strains of v. vulnificus were tes ...200818177963
genetic analysis of spontaneous lactose-utilizing mutants from vibrio vulnificus.wild-type v. vulnificus cannot grow using lactose as the sole carbon source or take up the sugar. however, prolonged culture of this species in media containing lactose as the sole carbon source leads to the generation of a spontaneous lactose-utilizing (lu) mutant. this mutant showed strong beta- galactosidase activity, whereas the wild-type strain showed a barely detectable level of the activity. a mutant with a lesion in a gene homologous to the lacz of e. coli in the bacterium no longer show ...200718167454
detection and characterization of a functional insertion sequence, isvpa2, in vibrio parahaemolyticus.pcr analysis of the pandemic strain of vibrio parahaemolyticus, kx-v237 (total genome sequenced) showed a subculture where the size of the amplicons had increased. the purpose of this study was to analyze the mechanism of this change. we found a 1,243-bp dna sequence inserted in one of the pandemic marker genes in this strain. the inserted dna sequence possessed the genetic structures shared by insertion sequences (iss) of the is3 family. this is had 26-bp imperfect terminal inverted repeats (ir ...200818164873
efficiency of real-time polymerase chain reaction assay to detect vibrio vulnificus in seawater.the growth of vibrio vulnificus in an enriched culture of seawater during the summer in japan was monitored by a plating technique used as the culture method and a real-time polymerase chain reaction (pcr) assay as the molecular method. v. vulnificus was detected by the real-time pcr assay in the samples of august and september but not by the culture method. vibrio parahaemolyticus, however, was detected among all of the samples with both the culture method and real-time pcr assay. in the analys ...200818162438
a common virulence plasmid in biotype 2 vibrio vulnificus and its dissemination aided by a conjugal plasmid.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 plasmids: pr99 (68.4 kbp) in strain cect4999 and pc4602-1 (56.6 kb) and pc4602-2 (66.9 kb) in strain cect4602. plasmid pc4602-2 showed ...200818156267
[vibrio vulnificus pollution of imported frozen black tiger shrimps in japan].the ariake sea area of japan is endemic for vibrio vulnificus infection. v vulnificus was isolated from slime from tidal flats, seawater, and fish sea year-round as we reported previously. to identify new routes and factors of v vulnificus infection, we studied v. vulnificus pollution of imported frozen black tiger shrimps purchased from a fish market in kurume, fukuoka, japan. v. vulnificus was isolated from 9 of 100 tails (9%) of philippines products, 3 of 100 tails (3%) of indonesia products, ...200718095471
molecular cloning and sequence analysis of a novel zinc-metalloprotease gene from the salinivibrio sp. strain af-2004 and its extracellular expression in e. coli.in this work the first protease gene encoding a novel zinc-metalloprotease from the moderately halophilic bacterium salinivibrio sp. strain af-2004 has been cloned, sequenced and reported to the genbank. we have generated a library containing about 10,000 transformants whose screening yielded one clone harboring plasmid pbluescript with 3.6 kb inserted fragment (pbluesvp2) with positive caseinolytic activity. nucleotide sequence analysis of the selected clone revealed a single open reading frame ...200818093752
discrimination of gamma-irradiated and nonirradiated vibrio vulnificus by using real-time polymerase chain reaction.to develop a pcr strategy for vibrio vulnificus in irradiated foods.200818081775
low-density lipoprotein protects vibrio vulnificus-induced lethality through blocking lipopolysaccharide action.lipoprotein plays a role in the host defense against bacterial infection, and its serum level has been demonstrated to be an important prognosis factor of survival. we have previously demonstrated that ldl directly inactivates the hemolytic activity of vibrio vulnificus cytolysin (vvc) in vitro. the object of this study was therefore to examine whether the ldl-mediated inactivation of vvc leads to protection against lethal infection of v. vulnificus in vivo, using wild and vvc-deficient v. vulni ...200718059143
role of the virulence plasmid pr99 and the metalloprotease vvp in resistance of vibrio vulnificus serovar e to eel innate immunity.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 aided by a conjugal plasmid. journal of bacteriology, submitted for publication.]. in this study, we analyzed the role of this plasmid tog ...200818053741
identification and functional analysis of vibrio vulnificus smcr, a novel global regulator.recently, quorum sensing has been implicated as an important global regulator controlling the production of numerous virulence factors such as capsular polysaccharides in bacterial pathogens. the nucleotide and deduced amino acid sequences of smcr, a homolog of v. harveyi luxr identified from v. vulnificus atcc29307, were analyzed. the amino acid sequence of smcr from v. vulnificus was 72 to 92% similar to those of luxr homologs from vibrio spp. functions of smcr were assessed by the constructio ...200718051765
ivet-based identification of virulence factors in vibrio vulnificus mo6-24/o.vibrio vulnificus is an opportunistic pathogen that causes septicemia in humans. to identify the genes associated with its pathogenicity, in vivo expression technology (ivet) was used to select genes specifically expressed in a host, yet not significantly in vitro. random lacz-fusions in the genome of v. vulnificus strain mo6-24/o were constructed using an ivet vector, psg3, which is a suicide vector containing promoterless-aph and -lacz as reporter genes. a total of approximately 18,000 resulti ...200718051754
the virulence of vibrio vulnificus is affected by the cellular level of superoxide dismutase activity.the virulence of superoxide dismutase (sod) mutants of vibrio vulnificus, as tested by intraperitoneal injection into mice, decreases in the order of sodc mutant, soda mutant, and sodb mutant lacking cuznsod, mnsod, and fesod, respectively. the survival of sod mutants under superoxide stress also decreases in the same order. the virulence of soxr mutant, which is unable to induce mnsod in response to superoxide, is similar to that of the soda mutant, as the survival of the soxr mutant under supe ...200718051612
cadaverine protects vibrio vulnificus from superoxide stress.an electron paramagnetic resonance (epr) signal characteristic of the 5,5'-dimethyl-1-pyrroline-n-oxide (dmpo)-oh spin adduct, which is formed from the reaction of dmpo with superoxide radicals generated by xanthine oxidase-mediated reaction, was significantly reduced by the cadaverine or escherichia coli mn-containing superoxide dismutase (mnsod). likewise, cytochrome c reduction by superoxide was inhibited by cadaverine, and the inhibition level increased in proportion to the level of cadaveri ...200718051370
role of the metalloprotease vvp and the virulence plasmid pr99 of vibrio vulnificus serovar e in surface colonization and fish virulence.the virulence for eels of vibrio vulnificus biotype 2 serovar e (vse) is conferred by a plasmid that codifies ability to survive in eel serum and cause septicaemia. to find out whether the plasmid and the selected chromosomal gene vvp plays a role in the initial steps of infection, the vse strain cect4999, the cured strain ct218 and the vvp-deficient mutant ct201 (obtained in this work by allelic exchange) were used in colonization and virulence experiments. the eel avirulent biotype 1 (bt1) str ...200818028416
the crystal structure of a binary complex of two pseudopilins: epsi and epsj from the type 2 secretion system of vibrio vulnificus.type ii secretion systems (t2ss) translocate virulence factors from the periplasmic space of many pathogenic bacteria into the extracellular environment. the t2ss of vibrio cholerae and related species is called the extracellular protein secretion (eps) system that consists of a core of multiple copies of 11 different proteins. the pseudopilins, epsg, epsh, epsi, epsj and epsk, are five t2ss proteins that are thought to assemble into a pseudopilus, which is assumed to interact with the outer mem ...200818022192
a widespread deferoxamine-mediated iron-uptake system in vibrio vulnificus.vibrio vulnificus can use the standard iron chelator deferoxamine (desferal) for efficient iron-uptake via the specific receptor desa, which is encoded by desa. we investigated the ubiquity of the deferoxamine-mediated iron-uptake system in v. vulnificus strains and the potential risk of the system. by polymerase chain reaction (pcr), desa was found in 10 of 10 clinical strains and in 9 of 10 environmental strains, and their growth was stimulated by deferoxamine. by reverse-transcriptase pcr, de ...200718008234
vibrio vulnificus rtx toxin kills host cells only after contact of the bacteria with host cells.vibrio vulnificus causes acute cell death and a fatal septicaemia. in this study, we show that contact with host cells is a prerequisite to the acute cytotoxicity. we screened transposon mutants defective in the contact-dependent cytotoxicity. two mutants had insertions within two open reading frames in a putative rtx toxin operon, the rtxa1 or rtxd encoding an rtx toxin (4701 amino acids) or an abc type transporter (467 amino acids). an rtxa1 mutation resulted in a cytotoxicity defect, which wa ...200818005241
a chromogenic substrate culture plate for early identification of vibrio vulnificus and isolation of other marine vibrios.vibrio vulnificus infection can result in necrotizing fasciitis and sepsis, which have short latentcy periods and high mortality rates. thus, an easy and quick detection method is needed to improve the outcome. to distinguish v. vulnificus from other pathogens that cause necrotizing fasciitis, we developed a selective isolation culture agar plate (chromochecker vibrio agar-1; cva-1) for use in environmental monitoring and in the clinical setting. one hundred four strains of v. vulnificus, alread ...200718000289
population structures of two genotypes of vibrio vulnificus in oysters (crassostrea virginica) and seawater.vibrio vulnificus biotype 1 strains can be classified into two genotypes based on the pcr analysis of variations in the virulence-correlated gene (vcg). genotype has been correlated with human infection for 90% of isolates from human cases having the vcgc sequence type and 87% of environmental strains having the vcge variant. in this study we examined the dynamics of v. vulnificus populations and the distribution of the two genotypes recovered from oysters and surrounding estuarine wasters. anal ...200817993556
survival of and in situ gene expression by vibrio vulnificus at varying salinities in estuarine environments.the opportunistic human pathogen vibrio vulnificus survives in a wide range of ecological environments, which demonstrates its ability to adapt to highly variable conditions. survival and gene expression under various conditions have been extensively studied in vitro; however, little work has been done to evaluate this bacterium in its natural habitat. therefore, this study monitored the long-term survival of v. vulnificus in situ and simultaneously evaluated the expression of stress (rpos, rela ...200817993554
vibrio infections in louisiana: twenty-five years of surveillance 1980-2005.a total of 1,007 vibrio infections were reported to the infectious disease epidemiology department at the louisiana office of public heath, between 1980 and 2005. the most common were vibrio vulnificus (257 infections), vibrio parahemolyticus (249 infections), and vibrio cholerae non o1 (200 cases). other species were much less common. vibrio vulnificus infections, which are associated with consumption of raw seafood (particularly oysters) or contact with sea water, and severe immuno-suppression ...200717987958
phospholipase a as a potent virulence factor of vibrio vulnificus.vibrio vulnificus infection has attracted special interest because of its high mortality rate. however, the identification of its major pathogenic determinant still remains obscure. in this study, a cytolysin-negative mutant strain of v. vulnificus cvd707 was used to determine the role of phospholipase a (pla) in the pathogenesis of this bacterial infection. the mutant strain caused the lysis of erythrocytes in vitro and elevated plasma hemoglobin during the infection in mice. both the hemolytic ...200717982702
survival case of vibrio vulnificus infection with multiple endocrine neoplasia type i.a 35-year-old female with malabsorption syndrome who underwent a pancreatoduodenectomy for multiple endocrine adenomatosis 13 years prior was admitted to our hospital with diarrhea, general fatigue, high fever, and eruption in the lower legs. the patient had consumed raw shrimp a few days before onset and presented systemic inflammatory response syndrome at the time of hospitalization. vibrio vulnificus was isolated from a blood culture performed before admission to the intensive care unit. we e ...200717974078
role of iron in human serum resistance of the clinical and environmental vibrio vulnificus genotypes.we recently reported a simple pcr procedure that targets a sequence variation of the virulence-correlated gene locus vcg. it was found that 90% of all clinical isolates possessed the vcgc sequence variant, while 93% of all environmental isolates possessed the vcge sequence variant. here we report that the clinical genotype of vibrio vulnificus is significantly better able to survive in human serum than is the environmental genotype. the presence of a siderophore-encoding gene, viub, influenced s ...200717933938
expression and immunogenicity analysis of two iron-regulated outer membrane proteins of vibrio parahaemolyticus.genes of two iron-regulated outer membrane proteins of vibrio parahaemolyticus zj2003, a pathogenic strain isolated from large yellow croaker (pseudosciaena crocea), psua and pvua, were cloned and expressed as n-terminal his(6)-tagged proteins in escherichia coli bl(21)(de(3)). the recombinant fusion proteins were purified with nickel chelate affinity chromatography. to analyze the immunogenicity of the proteins, groups of large yellow croaker were immunized with the purified recombinant psua, p ...200717928925
identification of a wzy polymerase required for group iv capsular polysaccharide and lipopolysaccharide biosynthesis in vibrio vulnificus.the estuarine bacterium vibrio vulnificus is a human and animal pathogen. the expression of capsular polysaccharide (cps) is essential for virulence. we used a new mini-tn10 delivery vector, pnktxi-scei, to generate a mutant library and identify genes essential for cps biosynthesis. twenty-one acapsular mutants were isolated, and the disrupted gene in one mutant, coding for a polysaccharide polymerase (wzy), is described here. a weca gene initiating glycosyltransferase was among the genes identi ...200717923517
evaluation of postharvest-processed oysters by using pcr-based most-probable-number enumeration of vibrio vulnificus bacteria.postharvest processing (php) is used to reduce levels of vibrio vulnificus in oysters, but process validation is labor-intensive and expensive. therefore, quantitative pcr was evaluated as a rapid confirmation method for most-probable-number enumeration (qpcr-mpn) of v. vulnificus bacteria in php oysters. qpcr-mpn showed excellent correlation (r(2) = 0.97) with standard mpn and increased assay sensitivity and efficiency.200717905883
comparing antibiotic resistance in commensal and pathogenic bacteria isolated from wild-caught south carolina shrimps vs. farm-raised imported shrimps.the objective of this study was to assess and differentiate wild-caught south carolina (sc) shrimps from imported shrimps on the basis of microbiological analysis. seven wild-caught sc shrimp and 13 farm-raised imported shrimp samples were analyzed. total plate counts from wild-caught shrimp samples ranged from 4.3 to 7.0 log10 cfu/g, whereas counts from imported shrimp samples ranged from 3.2 to 5.7 log10 cfu/g. there was no difference (p > 0.05) between total bacterial counts of wild-caught sc ...200717898847
the crucial amino acid residue related to inactivation of vibrio vulnificus hemolysin.vibrio vulnificus, an opportunistic human pathogen causing fetal septicemia, produces a 50-kda pore-forming toxin as a virulence factor. this toxin consists of 451 amino acid residues; however, there are two types of this toxin on the basis of the difference of some amino acid residues, type 1 (leu(281), ser(415), asn(435)/asp(435), asn(438)) and type 2 (ile(281), asn(415), asn(435), thr(438)). in the present study, two characteristic properties of type 2 toxin that was elaborated by v. vulnific ...200817897806
a novel multiplex pcr for the identification of vibrio parahaemolyticus, vibrio cholerae and vibrio vulnificus.to establish a simple multiplex polymerase chain reaction (pcr) that will identify vibrio parahaemolyticus, vibrio cholerae and vibrio vulnificus.200717897378
typing of vibrio vulnificus strains by variability in their 16s-23s rrna intergenic spacer regions.amplification of the 16s-23s rdna spacer region (isr1) is a simple and rapid procedure for subtyping bacteria, especially those with several ribosomal operons including vibrio vulnificus. v. vulnificus contains nine ribosomal operons with four or five isr1 classes that differ in size and sequence. in the present study, 47 v. vulnificus strains of both shellfish and clinical origin were subtyped by their isr1 patterns using "universal" primers, which target conserved sequences located in the 16s ...200717883316
information from your family doctor. vibrio vulnificus infection: what you should know. 200717853629
vibrio vulnificus infection: diagnosis and treatment.vibrio vulnificus infection is the leading cause of death related to seafood consumption in the united states. this virulent, gram-negative bacterium causes two distinct syndromes. the first is an overwhelming primary septicemia caused by consuming raw or undercooked seafood, particularly raw oysters. the second is a necrotizing wound infection acquired when an open wound is exposed to warm seawater with high concentrations of v. vulnificus. most patients, including those with primary infection, ...200717853628
phosphate and carbon source regulation of alkaline phosphatase and phospholipase in vibrio vulnificus.in this study, the effects of phosphate concentration and carbon source on the patterns of alkaline phosphatase (apase) and phospholipase (plase) expression in vibrio vulnificus atcc 29307 were assessed under various conditions. the activities of these enzymes were repressed by excess phosphate (4 mm) in the culture medium, but this repression was reversed upon the onset of phosphate starvation in low phosphate defined medium (lpdm) containing 0.2 mm of phosphate at approximately the end of the ...200717846584
antimicrobial susceptibilities of vibrio parahaemolyticus and vibrio vulnificus isolates from louisiana gulf and retail raw oysters.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 > or = 16 microg/ml). additionally, v. parahaemolyticus displayed significantly higher mics for cefotaxime, ciprofloxacin, and tetracycl ...200717827331
procaspase-3 activation by a metalloprotease secreted from vibrio vulnificus.vibrio vulnificus is a marine bacterium and a human pathogen capable of causing wound infection and septicemia. we previously showed that the metalloprotease vep secreted by v. vulnificus activates prothrombin in vitro. to further investigate the ability of vep to activate other zymogens, we used a mutant form of procaspase-3 which lacks the native cleavage sites as a zymogen. the mutant zymogen was activated by vep to yield a mature enzyme with a maximum increase in caspase-3 activity of approx ...200717786292
identification of a new hemolysin from diarrheal isolate ssu of aeromonas hydrophila.a clinical strain ssu of aeromonas hydrophila produces a potent cytotoxic enterotoxin (act) with cytotoxic, enterotoxic, and hemolytic activities. a new gene, which encoded a hemolysin of 439-amino acid residues with a molecular mass of 49 kda, was identified. this hemolysin (hlya) was detected based on the observation that the act gene minus mutant of a. hydrophila ssu still had residual hemolytic activity. the new hemolysin gene (hlya) was cloned, sequenced, and overexpressed in escherichia co ...200717725618
comparison of direct genome restriction enzyme analysis and pulsed-field gel electrophoresis for typing of vibrio vulnificus and their correspondence with multilocus sequence typing data.we compared the potential of direct genome restriction enzyme analysis (dgrea) and pulsed-field gel electrophoresis (pfge) for discriminating vibrio vulnificus isolates from clinical (23) and environmental (17) sources. the genotypes generated by both methodologies were compared to previous multilocus sequence typing (mlst) data. dgrea established clearer relationships among v. vulnificus strains and was more consistent with mlst than with pfge. dgrea is a very promising tool for epidemiological ...200717720834
the evolution of genetic structure in the marine pathogen, vibrio vulnificus.multi-locus sequence types (mlst) from a global collection of vibrio vulnificus isolates were analysed for the contribution of recombination to the evolution of two divergent clusters of strains and a human-pathogenic hybrid genotype, which caused a disease outbreak in israel. recombination contributes more substantially than mutation to generating strain diversity. for allelic diversity within loci, the ratio of recombination to mutation events is approximately 2:1. the role of recombination re ...200717716955
necrotizing fasciitis caused by vibrio vulnificus: epidemiology, clinical findings, treatment and prevention.necrotizing fasciitis is a soft-tissue infection with a high risk of fatality. infection with vibrio vulnificus can lead to development of necrotizing fasciitis and primary septicemia, and occurs mostly in immunocompromised host-associated diseases such as hepatic disease, diabetes mellitus, chronic renal insufficiency, and adrenal insufficiency. early recognition and treatment of the infection, which are unclear, are vital to patient welfare. we studied the disease epidemiology and reviewed the ...200717674061
[pathogenic vibrios in oysters (crassostrea rhizophorae) served at restaurants in rio de janeiro: a public health warning].forty oyster samples (crassostrea rhizophorae) served raw in 15 restaurants in the city of rio de janeiro were evaluated in order to investigate the presence of vibrio spp. the oyster samples were analyzed and subjected to enrichment in alkaline peptone water with the addition of 1 and 3% nacl and incubated at 37 degrees c for 24 hours. following this, the cultures were seeded onto thiosulfate citrate bile sucrose agar (tcbs) and the suspected colonies were subjected to biochemical characterizat ...200717653465
vibrio vulnificus typing based on simple sequence repeats: insights into the biotype 3 group.vibrio vulnificus is an opportunistic, highly invasive human pathogen with worldwide distribution. v. vulnificus strains are commonly divided into three biochemical groups (biotypes), most members of which are pathogenic. simple sequence repeats (ssr) provide a source of high-level genomic polymorphism used in bacterial typing. here, we describe the use of variations in mutable ssr loci for accurate and rapid genotyping of v. vulnificus. an in silico screen of the genomes of two v. vulnificus st ...200717652479
screening and evaluation of probiotics as a biocontrol agent against pathogenic vibrios in marine aquaculture.the present work aims at finding potential probionts from marine sources as a biocontrol agent against pathogenic vibrio species in shrimp larval culture.200717651222
the n-terminal propeptide of vibrio vulnificus extracellular metalloprotease is both an inhibitor of and a substrate for the enzyme.vibrio vulnificus, a marine bacterium capable of causing wound infection and septicemia, secretes a 45-kda metalloprotease (vep) with many biological activities. the precursor of vep consists of four regions: a signal peptide, an n-terminal propeptide (npp), a c-terminal propeptide, and the mature protease. two forms of vep-vep-45, which contains the mature protease plus the c-terminal propeptide, and vep-34, which contains only the mature protease-were expressed in escherichia coli and purified ...200717644589
vibrio vulnificus ilpa-induced cytokine production is mediated by toll-like receptor 2.vibrio vulnificus is a pathogenic bacterium causing primary septicemia, which follows a classical septic shock pathway, including an overwhelming inflammatory cytokine response. in this study, we identified a putative lipoprotein of v. vulnificus, encoded by the ilpa gene, as one of the surface proteins that specifically reacted with the antibodies raised against outer membrane proteins of v. vulnificus. using a mutant v. vulnificus in which its ilpa gene was knocked out, we found that ilpa is i ...200717640874
classification of response regulators based on their surface properties.the two-component signal transduction system is a ubiquitous signaling module present in most prokaryotic and some eukaryotic systems. two conserved components, a histidine protein kinase (hpk) protein and a response regulator (rr) protein, function as a biological switch, sensing and responding to changes in the environment, thereby eliciting a specific response. extensive studies have classified the hpk and rr proteins using primary sequence characteristics, domain identity, domain organizatio ...200717628138
phylogenetic diversity and the structural basis of substrate specificity in the beta/alpha-barrel fold basic amino acid decarboxylases.the beta/alpha-barrel fold type basic amino acid decarboxylases include eukaryotic ornithine decarboxylases (odc) and bacterial and plant enzymes with activity on l-arginine and meso-diaminopimelate. these enzymes catalyze essential steps in polyamine and lysine biosynthesis. phylogenetic analysis suggests that diverse bacterial species also contain odc-like enzymes from this fold type. however, in comparison with the eukaryotic odcs, amino acid differences were identified in the sequence of the ...200717626020
vibrio vulnificus infection in são paulo, brazil: case report and literature review.non cholera vibrio may cause conjunctivitis, wound infection, gastroenteritis and serious sepsis. transmission to men is through contact with skin, mucosa or wounds exposed to marine water, and consumption of certain barely cooked or raw seafood, more frequently in the summer. this is one of the first cases of severe infection related to vibrio vulnificus described in brazil. the patient was an old man, who ingested seafood in guarujá, a seashore city near são paulo, 3 days before hospitalizatio ...200717625784
inhibition of vibrio biofilm formation by a marine actinomycete strain a66.china remains by far the largest aquaculture producer in the world. however, biofilms formed by pathogenic vibrio strains pose serious problems to marine aquaculture. to provide a strategy for biofilm prevention, control, and eradication, extracts from 88 marine actinomycetes were screened. thirty-five inhibited the biofilm formation of vibrio harveyi, vibrio vulnificus, and vibrio anguillarum at a concentration of 2.5% (v/v). thirty-three of the actinomycete extracts dispersed the mature biofil ...200717624525
emergence of a virulent clade of vibrio vulnificus and correlation with the presence of a 33-kilobase genomic island.vibrio vulnificus is a ubiquitous inhabitant of the marine coastal environment, and an important pathogen of humans. we characterized a globally distributed sample of environmental isolates from a range of habitats and hosts and compared these with isolates recovered from cases of human infection. multilocus sequence typing data using six housekeeping genes divided 63 of the 67 isolates into the two main lineages previously noted for this species, and this division was also confirmed using the 1 ...200717616611
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