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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
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
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
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
diagnosing and treating deadly vibrio vulnificus infection. 200818358354
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
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
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
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
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
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
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
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
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
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
vibrio vulnificus and v. parahaemolyticus necrotising fasciitis in fishermen visiting an estuarine tropical northern australian location.three cases of severe necrotising fasciitis due to vibrio vulnificus (two cases) and vibrio parahaemolyticus (one case, fatal), have occurred in caucasian tourists while fishing at a remote tropical northern australian estuarine area. infections were acquired over a 4-year period during the tourist fishing season (april to july 2000-2003), when water temperatures range from 23 to 30 degrees c. they are notable for their geographical clustering in the remote western aspect of the gulf of carpenta ...200716890991
a real-time pcr assay for the rapid determination of 16s rrna genotype in vibrio vulnificus.in a terminal restriction fragment polymorphism (t-rflp) study, we recently reported a significant association between the type b 16s rrna gene and clinical strains of vibrio vulnificus associated with the consumption of raw oysters. in the present study we describe a real-time pcr assay for the rapid determination of the 16s rrna type of v. vulnificus isolates. this assay was used to reexamine the 16s rrna gene type in the strains studied previously by t-rflp and additional isolates from select ...200717070612
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
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
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
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
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
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
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
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
vibrio vulnificus peritonitis after eating raw sea fish in a patient undergoing continuous ambulatory peritoneal dialysis (capd). 200717205959
supra dietary levels of vitamins c and e enhance antibody production and immune memory in juvenile milkfish, chanos chanos (forsskal) to formalin-killed vibrio vulnificus.juveniles of milkfish, chanos chanos (forsskal), were fed two independent supra dietary levels of vitamins c (500 and 1500 mg kg(-1) feed, t1 and t2) and e (50 and 150 mg kg(-1), t3 and t4). milkfish fed diets with supra (in addition to the vitamins present in the control diet) and normal levels (t5 containing 90 and 1.2mg of vitamins c and e, respectively, kg(-1) of feed) of vitamins were immunized (ip) with formalin-killed vibrio vulnificus (fkvv). priming and booster antibody responses to the ...200717208456
the insect metalloproteinase inhibitor gene of the lepidopteran galleria mellonella encodes two distinct inhibitors.the insect metalloproteinase inhibitor (impi) from the greater wax moth, galleria mellonella, represents the first and to date only specific inhibitor of microbial metalloproteinases reported from animals. here, we report on the characterization including carbohydrate analysis of two recombinant constructs encoded by impi cdna either upstream or downstream of the furin cleavage site identified. rimpi-1, corresponding to native impi purified from hemolymph, is encoded by the n-terminal part of th ...200717214556
simultaneous flow cytometric assessment for cellular types and phagocytic abilities of the haemocytes of the hard clam, meretrix lusoria.the aim of this study was to devise a simple protocol for flow cytometric analysis to separate various haemocytic populations of the hard clam meretrix lusoria based on the mitochondrial membrane potential diversity detected by the fluorescence probe 3,3-dihexyloxacarbocyanine iodide (dioc6). compared with the traditional technique for separation of haemocytic populations, continuous percoll gradient centrifugation, our novel method was more efficient and yielded a higher ratio in separating the ...200717218115
first description of nonmotile vibrio vulnificus strains virulent for eels.nonmotile vibrio vulnificus strains were isolated as pure cultures from body ulcers and internal organs of wild diseased european eels caught in a mediterranean freshwater coastal lagoon. all 54 v. vulnificus isolates were nonmotile, indole-, ornithine decarboxilase-, mannitol- and cellobiose-positive, developed the opaque variant in culture, belonged to the o-antigenic serovar a and were highly virulent for eels by both intraperitoneal injection and immersion challenges. the nonmotile phenotype ...200717233719
positive regulation of fur gene expression via direct interaction of fur in a pathogenic bacterium, vibrio vulnificus.in pathogenic bacteria, the ability to acquire iron, which is mainly regulated by the ferric uptake regulator (fur), is essential to maintain growth as well as its virulence. in vibrio vulnificus, a human pathogen causing gastroenteritis and septicemia, fur gene expression is positively regulated by fur when the iron concentration is limited (h.-j. lee et al., j. bacteriol. 185:5891-5896, 2003). footprinting analysis revealed that an upstream region of the fur gene was protected by the fur prote ...200717237166
role of ntrc in biofilm formation via controlling expression of the gene encoding an adp-glycero-manno-heptose-6-epimerase in the pathogenic bacterium, vibrio vulnificus.to identify the genetic elements required for biofilm formation, we screened a pool of random vibrio vulnificus mutants for their ability to form biofilms. one mutant displaying significantly decreased biofilm-forming activity was found to contain a transposon insertion in the ntrc gene. the ntrc gene encodes a well-known transcriptional activator. we examined how this regulator modulates a biofilm-forming process in v. vulnificus by searching for ntrc target gene(s). comparison of the proteomes ...200717241201
necrotizing fasciitis from vibrio vulnificus in a patient with undiagnosed hepatitis and cirrhosis.necrotizing fasciitis due to vibrio vulnificus may result in overwhelming sepsis, leading to death in some patients. significant risk factors for severe disease include preexisting liver disease. we report a case of vibrio vulnificus necrotizing fasciitis in a patient with previously undiagnosed chronic hepatitis and cirrhosis.200717251404
vibrio vulnificus metalloprotease vvpe is essentially required for swarming.bacterial swarming constitutes a good in vitro model for surface adherence and colonization, and is accompanied by expressions of virulence factors related to invasiveness. in this study, it was determined that vibrio vulnificus swarming was abolished by mutation of the vvpe gene encoding a metalloprotease vvpe and this swarming defect was recovered by complementation of the vvpe gene. expression of the vvpe gene began simultaneously with the beginning of swarming and increased along with expres ...200717263846
vibrio vulnificus-necrotizing fasciitis in patient with cirrhosis. 200717268133
multiplex pcr assay for detection of vibrio vulnificus biotype 2 and simultaneous discrimination of serovar e strains.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.200717277209
membrane cholesterol is required for activity of vibrio vulnificus cytolysin.vibrio vulnificus cytolysin (vvc) forms a pore in the plasma membrane and induces cytolysis of various cells including erythrocytes, neutrophil and endothelial cells. the cytolytic activity of vvc is inhibited by exogenously added cholesterol, suggesting that membrane cholesterol might be required for vvc cytolytic activity. however, there is no direct evidence that membrane cholesterol is involved in vvc-induced cytolysis. herein we demonstrate that membrane cholesterol is required for binding ...200717285326
purification and characterization of a lectin from the banana shrimp fenneropenaeus merguiensis hemolymph.a lectin from the hemolymph of the banana shrimp fenneropenaeus merguiensis was purified by affinity chromatography on a fetuin-agarose column following by gel filtration on a superose-12 column. the native molecular mass of purified f. merguiensis lectin (fml) determined by gel filtration was 316.2 kda and its carbohydrate content was estimated to be 4.4%. by sds-page analysis, purified fml consisted of 32.3 kda and 30.9 kda subunits. these data suggest that this lectin is an oligomer. two-dime ...200716934939
osborn wave in hypothermia from vibrio vunificus sepsis unrelated to exposure.a 58 year old man with history of cirrhosis presented with vibrio vunificus sepsis. the patient developed multiorgan failure despite appropriate antibiotic therapy and fluid resuscitation. the patient developed moderate hypothermia. electrocardiography showed osborn wave. osborn wave is commonly seen in accidental hypothermia. although sepsis is known to cause hypothermia, osborn wave in sepsis is not widely appreciated.200717046085
rapid separation and concentration of food-borne pathogens in food samples prior to quantification by viable-cell counting and real-time pcr.buoyant density gradient centrifugation has been used to separate bacteria from complex food matrices, as well as to remove compounds that inhibit rapid detection methods, such as pcr, and to prevent false-positive results due to dna originating from dead cells. applying a principle of buoyant density gradient centrifugation, we developed a method for rapid separation and concentration following filtration and low- and high-speed centrifugation, as well as flotation and sedimentation buoyant den ...200717056684
identification of vibrio isolates by a multiplex pcr assay and rpob sequence determination.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 identifi ...200717093013
vibrio extracellular protease with prothrombin activation and fibrinolytic activities.a 36 kda extracellular metalloprotease (designated to as vep-mo6) was purified and characterized from vibrio vulnificus sp. strain mo6 24/0. vep-mo6 cleaved azocasein and a few other proteins such as prothrombin, plasminogen, fibrinogen and factor xa, which are associated with the blood coagulation pathway. the enzyme activity of vep-mo6 was inhibited by edta, which was reversed by the addition of excess divalent cations. vep-mo6 showed little or no activity toward various chromogenic substrates ...200717143560
ferrophilic characteristics of vibrio vulnificus and potential usefulness of iron chelation therapy.we determined the ferrophilic characteristics of vibrio vulnificus to evaluate the potential usefulness of iron chelation therapy for the prevention of v. vulnificus infection. readily available non-transferrin-bound iron (ntbi) is required for the initiation of v. vulnificus growth under in vitro iron-limited conditions and human ex vivo conditions. ntbi aided efficient transferrin-bound iron (tbi) use by v. vulnificus, and the vulnibactin-mediated iron-uptake system was expressed after bacteri ...200717152012
rapid detection of vibrio species using liquid microsphere arrays and real-time pcr targeting the ftsz locus.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 the ...200717172518
structural basis for sequence-dependent dna cleavage by nonspecific endonucleases.nonspecific endonucleases hydrolyze dna without sequence specificity but with sequence preference, however the structural basis for cleavage preference remains elusive. we show here that the nonspecific endonuclease cole7 cleaves dna with a preference for making nicks after (at 3'o-side) thymine bases but the periplasmic nuclease vvn cleaves dna more evenly with little sequence preference. the crystal structure of the 'preferred complex' of the nuclease domain of cole7 bound to an 18 bp dna with ...200717175542
isolation and characterization of a cold-induced nonculturable suppression mutant of vibrio vulnificus.the viable but nonculturable (vbnc) suppression mutant formed platable cells at low temperature stress after inoculation in artificial seawater (asw). suppression subtractive hybridization was used to identify differentially expressed genes among cdnas of the vbnc suppression mutant and the wild-type vibrio vulnificus strain. glutathione s-transferase was identified as a responsive gene of the vbnc suppression mutant in our assay, and was highly expressed from the vbnc suppression mutant at low ...200716520028
climate change and the emergence of vibrio vulnificus disease in israel.in 1996, a major unexplained outbreak of systemic vibrio vulnificus infection erupted among israeli fish market workers. the origins of this emergent infectious disease have not been fully understood. a possible link between climate change and disease emergence is being investigated. meteorological service data from 1981, the earliest detection and reporting of v. vulnificus for the time in israel, to 1998 for two stations located within the main inland fish farm industry were analyzed. the 1996 ...200716949069
serological and molecular characteristics of vibrio vulnificus biotype 3: evidence for high clonality.vibrio vulnificus biotype 3 has been implicated as the causative pathogen of an ongoing disease outbreak that erupted in israel in 1996. recent work based on multi-locus sequence typing (mlst) showed that v. vulnificus biotype 3 is genetically homogeneous. the aim of this study was to investigate the existence of subpopulations within this homogeneous biotype by characterizing the surface antigens and analysing the sequence diversity of selected outer-membrane protein (omp)-encoding genes. rabbi ...200717322205
shewanella infection in decompensated liver disease: a septic case.shewanella species are an unusual cause of disease in humans. however, reports of shewanella infections have been increasing, and hepatobiliary disease has been proposed as a predisposing factor following a critical course. we report the first japanese septic case of decompensated liver disease in which this bacterium acted as a definite pathogen. a 67-year-old japanese man with primary sclerosing cholangitis was admitted to our hospital complaining of fever, general fatigue, pain, and a rash on ...200717322999
occurrence of vibrio spp. in blue mussels (mytilus edulis) from the german wadden sea.the present study aimed at determining the contamination of blue mussels grown in germany with relevant bacterial organisms. seasonal variations were examined. for that purpose, mussel samples were taken regularly between june 2004 and may 2005 in seven shellfish-growing areas of the german wadden sea. a total of 90 samples were analysed. the analysis included the compulsory microbiological parameters escherichia coli and salmonella spp., as well as the determination of the contamination with vi ...200717331605
necrotizing soft-tissue infections and sepsis caused by vibrio vulnificus compared with those caused by aeromonas species.vibrio and aeromonas species, which can cause necrotizing fasciitis and primary septicemia, are members of the vibrionaceae family and thrive in aquatic environments. because the clinical symptoms and signs of necrotizing fasciitis and sepsis caused by these two bacteria are similar, the purposes of this study were to describe the clinical characteristics of vibrio vulnificus and aeromonas infections, to analyze the risk factors for death, and to compare the effects of surgical treatment on the ...200717332113
refined medium for direct isolation of vibrio vulnificus from oyster tissue and seawater.we have developed a new medium for the direct isolation of vibrio vulnificus from water and oyster samples. the medium was shown in laboratory and field studies to be highly selective without providing preferential isolation of either v. vulnificus genotype.200717337558
[studies on vibrio vulnificus infection: molecular epidemiology of environment-derived strains and clinical isolates].to clarify the route and source of vibrio vulnificus infection, we conducted molecular epidemiological investigation by dna analysis of 355 environmental isolates (seawater-derived strain: 86, sea mud-derived strain:36, and oyster-derived strain: 233) and 65 human clinical isolates, for a total of 420 isolates, using pulse field gel electrophoresis (pfge), with the following results. 1. when dna was cleaved with 2 enzymes, not i and sfi i, and subjected to pfge, not i dna interpretation was 76.9 ...200717338316
comment on "vibrio vulnificus and v. parahaemolyticus necrotising fasciitis in fishermen visiting an estuarine tropical northern australia location". 200717350104
chromosomal toxin-antitoxin loci can diminish large-scale genome reductions in the absence of selection.superintegrons (sis) are chromosomal genetic elements containing assemblies of genes, each flanked by a recombination sequence (attc site) targeted by the integron integrase. sis may contain hundreds of attc sites and intrinsic instability is anticipated; yet sis are remarkably stable. this implies that either selective pressure maintains the genes or mechanisms exist which favour their persistence in the absence of selection. toxin/antitoxin (ta) systems encode a stable toxin and a specific, un ...200717367382
the pyrh gene of vibrio vulnificus is an essential in vivo survival factor.we have suggested an important role of the pyrh gene during the infectious process of vibrio vulnificus. previously, we have identified 12 genes expressed preferentially during human infections by using in vivo-induced antigen technology. among the in vivo-expressed genes, pyrh encodes ump kinase catalyzing ump phosphorylation. introduction of a deletion mutation to the pyrh gene was lethal to v. vulnificus, and an insertional mutant showed a high frequency of curing. we constructed a site-direc ...200717371864
thermal factors influencing detection of vibrio vulnificus using real-time pcr.five thermal factors, including initial denaturation temperature, cycling denaturation temperature, annealing temperature, extension temperature and the temperature at which the intensity of the fluorescent signal is read, were evaluated for their effects on the detection of vibrio vulnificus via real-time pcr. fluorescent signal detection after extension was set between the tm value of the primer-dimers (79 degrees c) and that of the pcr target amplicons (84 degrees c). this effectively elimina ...200717383036
growth response of vibrio cholerae and other vibrio spp. to cyanobacterial dissolved organic matter and temperature in brackish water.environmental control of growth and persistence of vibrios in aquatic environments is poorly understood even though members of the genus vibrio are globally important pathogens. to study how algal-derived organic matter and temperature influenced the abundance of different vibrio spp., baltic sea microcosms inoculated with vibrio cholerae, vibrio vulnificus, vibrio parahaemolyticus, vibrio alginolyticus and native bacterioplankton, were exposed to different temperatures (12-25 degrees c) and ame ...200717386033
surface properties of vibrio vulnificus.vibrio vulnificus adheres to a diverse range of surfaces, ranging from the chitinous exoskeleton of mollusks to human tissue. to determine whether environmental and human clinical isolates exhibit different adhesion traits, we studied the ability of 10 environmental isolates and 10 clinical isolates to adhere to human epithelial cells and hydrocarbons with log p values ranging from 3.1 to 8.2.200717397482
recovery and detection of vibrio vulnificus during cold storage.different cultural techniques and molecular methods for the detection of vibrio vulnificus during cold storage in a model broth system were compared. two strains of v. vulnificus were grown to stationary phase and inoculated (10(6) cfu/ml) into tryptic soy broth with 2% sodium chloride (tsbn2) or artificial seawater (asw), both pre-chilled to 5 degrees c. these were stored for 10 days, with sub-sampling conducted at time 0 and every 2 days thereafter. each subsample was plated, by both pour and ...200717418319
the hlyu protein is a positive regulator of rtxa1, a gene responsible for cytotoxicity and virulence in the human pathogen vibrio vulnificus.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 ...200717438022
ligand-induced folding of the adenosine deaminase a-riboswitch and implications on riboswitch translational control.by using a structure-based fluorescence spectroscopic approach, we have examined the folding of an adenine-responsive riboswitch that regulates translation initiation. we observed adaptive recognition of the ligand for the aptamer domain of adenosine deaminase (add) mrna from vibrio vulnificus, and revealed pronounced conformational changes even in the preorganized loop-loop region that is distant from the binding site. importantly, the full-length riboswitch domain, which has a potential transl ...200717440909
synergistic antimicrobial effect of cefotaxime and minocycline on proinflammatory cytokine levels in a murine model of vibrio vulnificus infection.vibrio vulnificus causes primary bacteremia and necrotizing wound infection, leading to high morbidity and mortality in humans. this study aimed to evaluate the antimicrobial effect of cefotaxime and minocycline on proinflammatory cytokine levels in a murine model of v. vulnificus infection.200717446960
production of vibrio vulnificus metalloprotease vvpe begins during the early growth phase: usefulness of gelatin-zymography.recent studies have demonstrated that expression of the vvpe gene begins during the early growth phase albeit at low levels. however, we found that the traditional protease assay method that is used to measure caseinolytic activity in culture supernatants is not suitable for the measurement of extracellular vvpe that is produced at low levels during the early growth phase. by using gelatin-zymography in place of the protease assay, we could specifically detect only vvpe of several proteases prod ...200717467832
a short-chain dehydrogenase/reductase from vibrio vulnificus with both blue fluorescence and oxidoreductase activity.a dual function blue fluorescent protein from vibrio vulnificus is also an nadph-dependent oxidoreductase, rendering it a useful tool for biophysical studies.200717476406
identification and characterization of the vibrio vulnificus rtxa essential for cytotoxicity in vitro and virulence in mice.a mutant exhibiting decreased cytotoxic activity toward int-407 intestinal epithelial cells and carrying a mutation in the rtx gene cluster that consists of rtxca and rtxbde operons was screened from a library of v. vulnificus mutants. the functions of the rtxa gene, assessed by constructing an isogenic mutant and evaluating its phenotypic changes, demonstrated that rtxa is essential for the virulence of v. vulnificus in mice as well as in tissue cultures.200717483800
identification of quorum sensing-related regulons in vibrio vulnificus by two-dimensional gel electrophoresis and differentially displayed reverse transcriptase pcr.vibrio vulnificus is thought to employ a quorum-sensing system to control the expression of a global gene. in this study, proteomes and transcriptomes of a lacz null mutant, vvsr delta z, and a luxs-smcr double mutant, vvsr delta zsr, were compared with the parent strain, vvar, by means of two-dimensional gel electrophoresis (2d-page) and differentially displayed reverse transcriptase pcr (ddrt-pcr). 2d-page analysis showed that 36 protein spots were differentially expressed, 14 of which have be ...200717506728
phenotypic and genotypic characterization of a new fish-virulent vibrio vulnificus serovar that lacks potential to infect humans.vibrio vulnificus is a bacterial species that is virulent for humans and fish. human isolates are classified into biotypes 1 and 3 (bt1 and bt3) and fish isolates into biotype 2 (bt2). however, a few human infections caused by bt2 isolates have been reported worldwide (zoonosis). these bt2 human isolates belong to serovar e (sere), which is also present in diseased fish. the aim of the present work was to characterize a new bt2 serovar [serovar a (sera)], which emerged in the european fish-farmi ...200717526849
[isolation and identification of species from the genera aeromonas, vibrio, and plesiomonas from extraintestinal samples in cuba].extraintestinal infections caused by the genera aeromonas, vibrio and plesiomonas have high morbidity and mortality rates in different areas of world. from january 2002 to december 2003, the national reference laboratory for acute diarrhoeal diseases of the pedro kourí tropical medicine institute received 95 gram-negative, facultative anaerobic, oxidase positive bacilli strains from different extraintestinal specimen (blood, ear exudates, infected wounds, conjunctive exudates, urine, and cathete ...200717554439
role of type iv pilins in persistence of vibrio vulnificus in crassostrea virginica oysters.vibrio vulnificus is part of the natural estuarine microflora and accumulates in shellfish through filter feeding. it is responsible for the majority of seafood-associated fatalities in the united states mainly through consumption of raw oysters. previously we have shown that a v. vulnificus mutant unable to express pild, the type iv prepilin peptidase, does not express pili on the surface of the bacterium and is defective in adherence to human epithelial cells (r. n. paranjpye, j. c. lara, j. c ...200717557854
gene expression and localization of the epinecidin-1 antimicrobial peptide in the grouper (epinephelus coioides), and its role in protecting fish against pathogenic infection.epinecidin-1 is an antimicrobial peptide and plays a vital role in protecting fish against pathogenic infection. as a mimic of a grouper epinecidin-1 peptide, it has tertiary structures that closely resemble those of pleurocidin found in the winter flounder (pleuronectes americanus). the tissue-specific, lipopolysaccharide (lps)-stimulation-specific, and poly(i):poly(c)-stimulation-specific expressions of the grouper (epinephelus coioides) epinecidin-1 antimicrobial peptide were determined using ...200717570764
catecholamine-induced stimulation of growth in vibrio species.to evaluate the effect of norepinephrine (ne) and related compounds on the growth of bacteria, we have examined the effect of the neuroendocrine hormone ne and related compounds on the growth of vibrio parahaemolyticus and other human-pathogenic vibrio species (vibrio cholerae, vibrio vulnificus, and vibrio mimicus).200717576228
vibrio vulnificus damages macrophages during the early phase of infection.vibrio vulnificus is an estuarine bacterium that can cause primary septicemia as well as serious wound infections. generally, clinical isolates have a high lethal effect compared with environmental isolates. however, little is known about the mechanisms by which v. vulnificus causes disease. in this study, we compared the pathogenicity of a clinical isolate, strain m2799, with that of an environmental isolate, strain jcm3731. the clinical isolate showed 100 times higher lethality in mice than th ...200717591793
pathogenic bacteria associated with oysters (crassostrea brasiliana) and estuarine water along the south coast of brazil.oysters and estuarine water samples were collected monthly, from june 1998 to march 1999, in the cananéia estuary, on the south coast of são paulo, brazil, and analyzed for bacterial hazards with and without depuration in filtered estuarine water. aeromonas spp., plesiomonas shigelloides, vibrio cholerae o1, vibrio parahaemolyticus, and vibrio vulnificus were counted in oyster samples using the most probable number (mpn) and their presence verified in the surrounding estuarine water samples. the ...200717613090
rapid and specific identification of 5 human pathogenic vibrio species by multiplex polymerase chain reaction targeted to dnaj gene.a multiplex polymerase chain reaction (pcr) method, specifically designed for application in routine diagnostic laboratories, was developed for identifying 5 human pathogen vibrio species: vibrio cholerae, vibrio parahaemolyticus, vibrio vulnificus, vibrio mimicus, and vibrio alginolyticus. this assay directed toward the dnaj gene was tested on a total of 355 strains representing 13 vibrio species and 17 non-vibrio species. specific pcr fragments were produced in isolates belonging to the 5 targ ...200717614235
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
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
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
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
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
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
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
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
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
[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
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
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
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
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
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
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
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
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
information from your family doctor. vibrio vulnificus infection: what you should know. 200717853629
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
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
[sepsis and cellulitis by vibrio vulnificus in cirrhotic patient]. 200718198957
[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
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
the growth of vibrio vulnificus and the habitat of infected patients in kumamoto.in japan, vibrio vulnificus (v. vulnificus) infection is very rare, and most infections have occurred in kumamoto prefecture (1), and especially around the ariake and yatsushiro seas. to investigate the relationship between the occurrence of v. vulnificus infection and environmental factors, including the salinity of seawater and the amount of rain in the ariake and yatsushiro seas, we measured the most probable number (mpn) of v. vulnificus in seawater and sea mud. in the ariake sea, we also ob ...200720103883
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