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Sample records for vibrio vulnificus isolated

  1. Indole-positive Vibrio vulnificus isolated from disease outbreaks on a Danish eel farm

    DEFF Research Database (Denmark)

    Dalsgaard, Inger; Høi, L.; Siebeling, R.J.

    1999-01-01

    Vibrio vulnificus was isolated in 1996 from 2 disease outbreaks on a Danish eel farm which used brackish water. A characteristic clinical sign was extensive, deep muscle necrosis in the head region. V. vulnificus was isolated from kidney, mucus, spleen, gill and intestine of diseased eels. Thirty...

  2. Heterogeneity among isolates of Vibrio vulnificus recovered from eels ( Anguilla anguilla ) in Denmark

    DEFF Research Database (Denmark)

    Høi, L.; Dalsgaard, Inger; DePaola, A.

    1998-01-01

    are genetically heterogenous as shown by ribotyping, Ribotyping also showed that the same isolates map persist in an eel farm and cause recurrent outbreaks, Phage typing did not correlate with ribotyping or serotyping. However, we observed that 26 of 28 isolates, which mere not susceptible to any of the phages......The findings of this study demonstrate that Vibrio vulnificus isolates recovered from diseased eels in Denmark are heterogeneous as shown by O serovars, capsule types, ribotyping, phage typing, and plasmid profiling, The study: includes 85 V. vulnificus isolates isolated from the gills, intestinal...

  3. Comparison of a commercial biochemical kit and an oligonucleotide probe for identification of environmental isolates of Vibrio vulnificus

    DEFF Research Database (Denmark)

    Dalsgaard, A.; Dalsgaard, Inger; Høi, L.

    1996-01-01

    Methods for the identification and isolation of environmental isolates of Vibrio vulnificus were evaluated. Alkaline peptone water supplemented with polymyxin B and colistin-polymyxin B- cellobiose agar were employed for the isolation of suspected V. vulnificus from water, sediment and shellfish...

  4. Autecology of Vibrio vulnificus and Vibrio parahaemolyticus in tropical waters

    Energy Technology Data Exchange (ETDEWEB)

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

    1988-12-31

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

  5. Zoonose Vibrio vulnificus: meldingsplicht raadzaam

    NARCIS (Netherlands)

    Dijkstra, A.; Haenen, O.L.M.; Moller, L.

    2010-01-01

    Op de lijst van meldingsplichtige infectieziekten komen een aantal zoönosen voor, zoals pest, rabiës en leptospirose. De relatief onbekende zoönotische Vibrio vulnificus valt opmerkelijk genoeg niet onder de meldingsplichtige infectieziekten. Juist vanwege het zeer agressieve beloop van een

  6. Virulence Profiles of Vibrio vulnificus in German Coastal Waters, a Comparison of North Sea and Baltic Sea Isolates

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    Nadja Bier

    2015-12-01

    Full Text Available Vibrio vulnificus is a halophilic bacterium of coastal environments known for sporadically causing severe foodborne or wound infections. Global warming is expected to lead to a rising occurrence of V. vulnificus and an increasing incidence of human infections in Northern Europe. So far, infections in Germany were exclusively documented for the Baltic Sea coast, while no cases from the North Sea region have been reported. Regional variations in the prevalence of infections may be influenced by differences in the pathogenicity of V. vulnificus populations in both areas. This study aimed to compare the distribution of virulence-associated traits and genotypes among 101 V. vulnificus isolates from the Baltic Sea and North Sea in order to assess their pathogenicity potential. Furthermore, genetic relationships were examined by multilocus sequence typing (MLST. A high diversity of MLST sequences (74 sequence types and differences regarding the presence of six potential pathogenicity markers were observed in the V. vulnificus populations of both areas. Strains with genotypes and markers associated with pathogenicity are not restricted to a particular geographic region. This indicates that lack of reported cases in the North Sea region is not caused by the absence of potentially pathogenic strains.

  7. Comparative genomic analysis of clinical and environmental Vibrio vulnificus isolates revealed biotype 3 evolutionary relationships.

    Science.gov (United States)

    Koton, Yael; Gordon, Michal; Chalifa-Caspi, Vered; Bisharat, Naiel

    2014-01-01

    In 1996 a common-source outbreak of severe soft tissue and bloodstream infections erupted among Israeli fish farmers and fish consumers due to changes in fish marketing policies. The causative pathogen was a new strain of Vibrio vulnificus, named biotype 3, which displayed a unique biochemical and genotypic profile. Initial observations suggested that the pathogen erupted as a result of genetic recombination between two distinct populations. We applied a whole genome shotgun sequencing approach using several V. vulnificus strains from Israel in order to study the pan genome of V. vulnificus and determine the phylogenetic relationship of biotype 3 with existing populations. The core genome of V. vulnificus based on 16 draft and complete genomes consisted of 3068 genes, representing between 59 and 78% of the whole genome of 16 strains. The accessory genome varied in size from 781 to 2044 kbp. Phylogenetic analysis based on whole, core, and accessory genomes displayed similar clustering patterns with two main clusters, clinical (C) and environmental (E), all biotype 3 strains formed a distinct group within the E cluster. Annotation of accessory genomic regions found in biotype 3 strains and absent from the core genome yielded 1732 genes, of which the vast majority encoded hypothetical proteins, phage-related proteins, and mobile element proteins. A total of 1916 proteins (including 713 hypothetical proteins) were present in all human pathogenic strains (both biotype 3 and non-biotype 3) and absent from the environmental strains. Clustering analysis of the non-hypothetical proteins revealed 148 protein clusters shared by all human pathogenic strains; these included transcriptional regulators, arylsulfatases, methyl-accepting chemotaxis proteins, acetyltransferases, GGDEF family proteins, transposases, type IV secretory system (T4SS) proteins, and integrases. Our study showed that V. vulnificus biotype 3 evolved from environmental populations and formed a genetically

  8. Comparative Genomic Analysis of Clinical and Environmental Vibrio Vulnificus Isolates Revealed Biotype 3 Evolutionary Relationships

    Directory of Open Access Journals (Sweden)

    Yael eKotton

    2015-01-01

    Full Text Available In 1996 a common-source outbreak of severe soft tissue and bloodstream infections erupted among Israeli fish farmers and fish consumers due to changes in fish marketing policies. The causative pathogen was a new strain of Vibrio vulnificus, named biotype 3, which displayed a unique biochemical and genotypic profile. Initial observations suggested that the pathogen erupted as a result of genetic recombination between two distinct populations. We applied a whole genome shotgun sequencing approach using several V. vulnificus strains from Israel in order to study the pan genome of V. vulnificus and determine the phylogenetic relationship of biotype 3 with existing populations. The core genome of V. vulnificus based on 16 draft and complete genomes consisted of 3068 genes, representing between 59% and 78% of the whole genome of 16 strains. The accessory genome varied in size from 781 kbp to 2044 kbp. Phylogenetic analysis based on whole, core, and accessory genomes displayed similar clustering patterns with two main clusters, clinical (C and environmental (E, all biotype 3 strains formed a distinct group within the E cluster. Annotation of accessory genomic regions found in biotype 3 strains and absent from the core genome yielded 1732 genes, of which the vast majority encoded hypothetical proteins, phage-related proteins, and mobile element proteins. A total of 1916 proteins (including 713 hypothetical proteins were present in all human pathogenic strains (both biotype 3 and non-biotype 3 and absent from the environmental strains. Clustering analysis of the non-hypothetical proteins revealed 148 protein clusters shared by all human pathogenic strains; these included transcriptional regulators, arylsulfatases, methyl-accepting chemotaxis proteins, acetyltransferases, GGDEF family proteins, transposases, type IV secretory system (T4SS proteins, and integrases. Our study showed that V. vulnificus biotype 3 evolved from environmental populations and

  9. Vibrio vulnificus outbreaks in Dutch eel farms since 1996: strain diversity and impact.

    NARCIS (Netherlands)

    Haenen, O.L.M.; Zanten, van E.; Jansen, R.; Roozenburg, I.; Engelsma, M.Y.; Dijkstra, A.; Boers, S.A.; Voorbergen-Laarman, M.; Möller, A.V.M.

    2014-01-01

    Vibrio vulnificus is a potentially zoonotic bacterial pathogen of fish, which can infect humans (causing necrotic fasciitis). We analysed 24 V. vulnificus isolates (from 23 severe eel disease outbreaks in 8 Dutch eel farms during 1996 to 2009, and 1 clinical strain from an eel farmer) for genetic

  10. Prevalence of Vibrio vulnificus and Vibrio parahaemolyticus in the Maryland Coastal Bays

    Science.gov (United States)

    De Pascuale, V. O.

    2016-02-01

    The bacterial family of Vibrionaceae is indigenous in the marine estuarine environments such as the Maryland Coastal Bays. Vibrio vulnificus and Vibrio parahaemolyticus are both pathogenic bacteria. Understanding the distribution of Vibrio species is crucial because of the health concerns associated with the bacteria. The aim of this study was to evaluate the overall abundance of bacteria with a focus on Vibrio species in the Maryland Coastal Bays. Seawater samples were collected from 10 different sites that differ with regard to water quality. The total bacteria count (TBC) was determined by two methods: Total plate count and Epifluorescence microscopy. The most-probable-number (MPN) methodology was used to estimate the population of Vibrio parahaemolyticus and Vibrio vulnificus. In addition to the bacteriological analysis, the environmental parameters of temperature and salinity were measured using YSI 6600 multiparameter meter. The average total bacteria count was 2.21 log CFU ml-1. Vibrio vulnificus comprised 5% of the total bacteria count while Vibrio parahaemolyticus comprised only 2% of the total bacteria count. Vibrio vulnificus ranged from 0.30 to 2.48 log MPN ml-1 at the sites tested. Lower Vibrio parahaemolyticus count was observed at the sites with a range of 0.30 to 1.97 log MPN ml-1. There was no significant correlation between the environmental parameters and the Vibrio spp. Since both Vibrio vulnificus and Vibrio parahaemolyticus peak in the summer, there is a potential for a risk of wound infections and gastrointestinal illness based on this data.

  11. Vibrio vulnificus-infektioner i Danmark sommeren 1994

    DEFF Research Database (Denmark)

    Bruun, Brita Grønbech; Frimodt-Møller, N; Dalsgaard, A.

    1996-01-01

    The clinical manifestations and epidemiological data of 11 patients infected with Vibrio vulnificus found in Denmark during the unusually warm summer of 1994 are reported. All patients had been exposed to seawater prior to illness, but none had consumed seafood. Nine patients, including four...

  12. Vibrio vulnificus induces mTOR activation and inflammatory responses in macrophages.

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    Dan-Li Xie

    Full Text Available Vibrio vulnificus (V. vulnificus, a Gram-negative marine bacterium, can cause life-threatening primary septicemia, especially in patients with liver diseases. How V. vulnificus affects the liver and how it acts on macrophages are not well understood. In this report, we demonstrated that V. vulnificus infection causes a strong inflammatory response, marked expansion of liver-resident macrophages, and liver damage in mice. We demonstrated further that V. vulnificus activates mTOR in macrophages and inhibition of mTOR differentially regulates V. vulnificus induced inflammatory responses, suggesting the possibility of targeting mTOR as a strategy to modulate V. vulnificus induced inflammatory responses.

  13. Identification of DNA Sequences Specific for Vibrio vulnificus Biotype 2 Strains by Suppression Subtractive Hybridization

    OpenAIRE

    Lee, Chung-Te; Amaro, Carmen; Sanjuán, Eva; Hor, Lien-I

    2005-01-01

    Vibrio vulnificus can be divided into three biotypes, and only biotype 2, which is further divided into serovars, contains eel-virulent strains. We compared the genomic DNA of a biotype 2 serovar E isolate (tester) with the genomic DNAs of three biotype 1 strains by suppression subtractive hybridization and then tested the distribution of the tester-specific DNA sequences in a wide collection of bacterial strains. In this way we identified three plasmid-borne DNA sequences that were specific ...

  14. Role of iron, capsule, and toxins in the pathogenicity of Vibrio vulnificus biotype 2 for mice.

    OpenAIRE

    Amaro, C; Biosca, E G; Fouz, B; Toranzo, A E; Garay, E

    1994-01-01

    The virulence mechanisms of Vibrio vulnificus biotype 2 have been studied and compared with those of biotype 1 in mice as the experimental animals. Biotype 2 isolates from European eels were as virulent for mice as biotype 1 strains (50% lethal dose, about 10(5) CFU per mouse); a septicemic infection developed in less than 24 h. These strains had several properties in common with biotype 1 organisms including capsule expression, uptake of various iron sources, and production of exoproteins, w...

  15. Transmission to Eels, Portals of Entry, and Putative Reservoirs of Vibrio vulnificus Serovar E (Biotype 2)

    OpenAIRE

    Marco-Noales, Ester; Milán, Miguel; Fouz, Belén; Sanjuán, Eva; Amaro, Carmen

    2001-01-01

    Vibrio vulnificus serovar E (formerly biotype 2) is the etiologic agent that is responsible for the main infectious disease affecting farmed eels. Although the pathogen can theoretically use water as a vehicle for disease transmission, it has not been isolated from tank water during epizootics to date. In this work, the mode of transmission of the disease to healthy eels, the portals of entry of the pathogen into fish, and their putative reservoirs have been investigated by means of laborator...

  16. Morphological characterization and biocontrol effects of Vibrio vulnificus phages against Vibriosis in the shrimp aquaculture environment.

    Science.gov (United States)

    Srinivasan, Pappu; Ramasamy, Palaniappan

    2017-10-01

    The re-emerging field of phage therapy is the potential biocontrol agents for the transfer of virulence factor and to kill their bacterial hosts. In this study, the lytic Vibrio vulnificus phages were studied to provide a better understanding of phage-host interactions and development of phage therapy. Four new V. vulnificus phages were detected from shrimp aquaculture system, named VV1, VV2, VV3 and VV4. All lytic V. vulnificus phages are the Tectiviruses of the family Tectiviridae with typical double layered elongated icosahedral head and tailless morphology. Lytic V. vulnificus phages which infect other Vibrio isolates were further characterized by long term storage, enzymes treatment, organic solvents treatment, detergents treatment, pH stability, temperature stability, agar bioassay method and one-step growth experiment. The effects of chloroform, acetone, ethyl alcohol, methyl alcohol, ribonuclease (RNase), trypsin, protease, Triton-X100 treatments were not affected the growth of VV1, VV2, VV3 and VV4 phages. These phages (VV1-VV4) were inactivated completely with temperature (over 60 °C), pH (12), lysozyme and sodium dodecyl sulfate (SDS) treatment. One-step growth experiments indicated that the latent period was at 3 h and burst size was at 37 °C. Agar bioassay method indicated that the percentage of inhibition was 75% (VV1) and 70% (VV2, VV3 & VV4), respectively. SDS-PAGE analysis of all V. vulnificus phages had similar protein patterns with molecular weight masses of 260, 249, 204, 148, 63, 59, 22 and 15 kDa. Hence, it could be concluded that V. vulnificus phage had a broad lytic spectrum and potential biocontrol of luminous Vibriosis in the shrimp aquaculture system. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Prevalence and population structure of Vibrio vulnificus on fishes from the northern Gulf of Mexico.

    Science.gov (United States)

    Tao, Zhen; Larsen, Andrea M; Bullard, Stephen A; Wright, Anita C; Arias, Covadonga R

    2012-11-01

    The prevalence of Vibrio vulnificus on the external surfaces of fish from the northern Gulf of Mexico was determined in this study. A collection of 242 fish comprising 28 species was analyzed during the course of 12 sampling trips over a 16-month period. The prevalence of V. vulnificus was 37% but increased up to 69% in summer. A positive correlation was found between the percentages of V. vulnificus-positive fish and water temperatures, while salinity and V. vulnificus-positive fish prevalence were inversely correlated. A general lineal model (percent V. vulnificus-positive fish = 0.5930 - 0.02818 × salinity + 0.01406 × water temperature) was applied to best fit the data. Analysis of the population structure was carried out using 244 isolates recovered from fish. Ascription to 16S rRNA gene types indicated that 157 isolates were type A (62%), 72 (29%) were type B, and 22 (9%) were type AB. The percentage of type B isolates, considered to have greater virulence potential, was higher than that previously reported in oyster samples from the northern Gulf of Mexico. Amplified fragment length polymorphism (AFLP) was used to resolve the genetic diversity within the species. One hundred twenty-one unique AFLP profiles were found among all analyzed isolates, resulting in a calculated Simpson's index of diversity of 0.991. AFLP profiles were not grouped on the basis of collection date, fish species, temperature, or salinity, but isolates were clustered into two main groups that correlated precisely with 16S rRNA gene type. The population of V. vulnificus associated with fishes from the northern Gulf of Mexico is heterogeneous and includes strains of great virulence potential.

  18. Chitinase determinants of Vibrio vulnificus: gene cloning and applications of a chitinase probe.

    OpenAIRE

    Wortman, A T; Somerville, C C; Colwell, R R

    1986-01-01

    To initiate study of the genetic control of chitinolytic activity in vibrios, the chitobiase gene was isolated by cloning chromosomal DNA prepared from Vibrio vulnificus. Chimeric plasmids were constructed from Sau3A I partial digests of chromosomal DNA by ligating 5 to 15-kilobase fragments into the BamHI site, i.e., in the Tcr gene, of pBR322 (Amr Tcr). The resulting plasmids were transformed into Escherichia coli DH1. Chitobiase activity of the insert-bearing clones was detected by using a...

  19. Environmental occurrence and clinical impact of Vibrio vulnificus and Vibrio parahaemolyticus: a European perspective.

    Science.gov (United States)

    Baker-Austin, Craig; Stockley, Louise; Rangdale, Rachel; Martinez-Urtaza, Jaime

    2010-02-01

    Vibrio vulnificus and Vibrio parahaemolyticus are ubiquitous Gram-negative bacterial pathogens found naturally in marine and estuarine waters, and are a leading cause of seafood-associated bacterial illness. These pathogens are commonly reported in the USA and in many Asian countries, including China, Japan and Taiwan; however, there is growing concern that V. vulnificus and V. parahaemolyticus may represent an important and increasing clinical problem in Europe. Several factors underlie the need for a greater understanding of these non-cholera vibrios within a European context. First, there is a growing body of evidence to suggest that V. vulnificus and V. parahaemolyticus infections are increasing, and tend to follow regional climatic trends, with outbreaks typically following episodes of unusually warm weather. Such findings are especially alarming given current predictions regarding warming of marine waters as a result of global climatic change. Second, a myriad of epidemiological factors may greatly increase the incidence as well as clinical burden of these pathogens - including increasing global consumption and trade of seafood produce coupled to an increase in the number of susceptible individuals consuming seafood produce. Finally, there is currently a lack of detailed surveillance information regarding non-cholerae Vibrio infections in Europe, as these pathogens are not notifiable in many countries, which probably masks the true clinical burden of many human infections. This review will present a pertinent overview of both the environmental occurrence and clinical impact of V. vulnificus and V. parahaemolyticus in Europe. © 2010 Crown copyright.

  20. Insight into the evolution of Vibrio vulnificus biotype 3's genome.

    Science.gov (United States)

    Efimov, Vera; Danin-Poleg, Yael; Raz, Nili; Elgavish, Sharona; Linetsky, Alex; Kashi, Yechezkel

    2013-01-01

    Vibrio vulnificus is an aquatic bacterium and an important human pathogen. Strains of V. vulnificus are biochemically classified into three biotypes. The newly emerged biotype 3 appears to be rather clonal and geographically restricted to Israel, where it caused an outbreak of wound infections and bacteremia. To understand the evolution of the bacterium's genome, we sequenced and analyzed the genome of biotype 3 strain VVyb1(BT3), and then conducted a microbial environmental survey of the hypothesized niche from which it probably evolved. The genome of this environmental isolate revealed higher similarity to the published biotype 1 genomes of clinical strains (90%) than to the environmental strains (87%), supporting the virulence of the biotype 3 group. Moreover, 214 of the total 5361 genes were found to be unique to strain VVyb1(BT3), having no sequence similarity to any of the known genomes of V. vulnificus; 35 of them function in DNA mobility and rearrangement, supporting the role of horizontal gene transfer in genome evolution. Interestingly, 29 of the "unique" genes had homologies among Shewanella species. In a survey conducted in aquaculture ponds in Israel, we successfully co-isolated Shewanella and V. vulnificus from the same niche, further supporting the probable contribution of Shewanella to the genome evolution of biotype 3. Indeed, one gene was found in a S. algae isolate. Surprisingly, molecular analysis revealed that some of the considered unique genes are harbored by non-sequenced biotype 1 strains isolated from the same environment. Finally, analyses of the biotype 3 genome together with the environmental survey suggested that its genome originated from a biotype 1 Israeli strain that acquired a rather small number of genes from other bacterial species in the niche, such as Shewanella. Therefore, aquaculture is likely to play a major role as a man-made ecological niche in bacterial evolution, leading the emergence of new pathogenic groups in V

  1. Vibrio vulnificus infection in Southern Brazil - Case report Infecção por Vibrio vulnificus no sul do Brasil - Relato de caso

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    João César Beenke França

    2013-06-01

    Full Text Available The genus Vibrio is a member of the family Vibrionaceae, and among their disease-causing species, Vibrio vulnificus, a lactose-positive gram-negative bacillus, is one of the most virulent pathogen of the noncholerae vibrios. We describe the case of a 39-year-old male patient, who was using immunosuppressive therapy, admitted to the hospital for liver transplantation. Twelve hours later, the patient presented high fever, myalgia, anuria and erythematous plaques on lower limbs, of rapid growth and proximal progression. The patient was treated with ceftriaxone, meropenem and oxacillin, however he expired within 30 hours. Blood cultures showed growth of a gram-negative bacillus, which was later identified as Vibrio vulnificus.O gênero Vibrio é membro da família Vibrionaceae, e entre as espécies patogênicas, Vibrio vulnificus, bacilo gram negativo lactose positivo, tem sido frequentemente citado. Descrevemos o caso de um paciente masculino de 39 anos, em uso de medicação imunossupressora, admitido no hospital para transplante hepático. Doze horas após a internação, o paciente evoluiu com febre, mialgias, anúria e placas eritematosas em membros inferiores, com rápido crescimento e evolução proximal. O paciente foi tratado com ceftriaxona, meropenem e oxacilina sem melhora, evoluindo para óbito em 30 horas. Hemocultura mostrou crescimento de bacilo gram negativo posteriormente identificado como Vibrio vulnificus.

  2. Vibrio vulnificus infection in São Paulo, Brazil: case report and literature review

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    Maria Rita Elmor de Araujo

    Full Text Available 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 hospitalization. He was admitted to the emergency room in an ill state with septic shock. On 2 sets of blood culture a highly virulent microorganism was isolated, Vibrio vulnificus, which leads to sepsis and frequently to death in susceptible patients. The objective of this report was to use this case to discuss clinical aspects, microbiological diagnosis and treatment of the infection caused by this agent, besides the review of epidemiology, associated risk factors and prevention before consuming or getting in contact with seafood, especially in patients with greater susceptibility to this kind of infection.

  3. A case of Vibrio vulnificus infection complicated with fulminant purpura: gene and biotype analysis of the pathogen.

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    Hori, Masatoshi; Nakayama, Akifumi; Kitagawa, Daisuke; Fukushima, Hidetada; Asai, Hideki; Kawai, Yasuyuki; Okuchi, Kazuo

    2017-05-01

    Introduction.Vibrio vulnificus (V. vulnificus) causes a severe infection that develops in the compromised host. Its pathophysiology is classified into three types: (1) primary septicaemia, (2) gastrointestinal illness pattern and (3) wound infection pattern. Of these, primary septicaemia is critical. V. vulnificus can be classified into three biotypes and two genotypes and its pathogenicity is type-dependent. Case presentation. A 47-year-old man presented to a local hospital with chief complaints of fever, bilateral lower limb pain and diarrhoea. He had no history of foreign travel or known medical problems. He was in septic shock and developed fulminant purpura within 24 h of the onset. High-dose vasopressor and antibiotic administration failed to alter his status and he died 3 days after the onset of symptoms. V. vulnificus was isolated from blood, skin and nasal discharge cultures. Biotype and gene analysis of the microbe isolated identified it as Biotype 3, mainly reported in Israel in wound infections, and Genotype E, implicating an environmental isolate. These typing analyses indicated that the microbe isolated could be classified as a type with low pathogenicity. Conclusion. This case highlighted that Biotype 3 and Genotype E can also cause primary septicaemia. Although the majority of reports on Biotype 3 have been from the Middle East, this experience with the present case provided evidence that the habitat of Biotype 3 V. vulnificus has been extending to East Asia as well.

  4. Rapid proliferation of Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae during freshwater flash floods in French Mediterranean coastal lagoons.

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    Esteves, Kevin; Hervio-Heath, Dominique; Mosser, Thomas; Rodier, Claire; Tournoud, Marie-George; Jumas-Bilak, Estelle; Colwell, Rita R; Monfort, Patrick

    2015-11-01

    Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae of the non-O1/non-O139 serotype are present in coastal lagoons of southern France. In these Mediterranean regions, the rivers have long low-flow periods followed by short-duration or flash floods during and after heavy intense rainstorms, particularly at the end of the summer and in autumn. These floods bring large volumes of freshwater into the lagoons, reducing their salinity. Water temperatures recorded during sampling (15 to 24°C) were favorable for the presence and multiplication of vibrios. In autumn 2011, before heavy rainfalls and flash floods, salinities ranged from 31.4 to 36.1‰ and concentrations of V. parahaemolyticus, V. vulnificus, and V. cholerae varied from 0 to 1.5 × 10(3) most probable number (MPN)/liter, 0.7 to 2.1 × 10(3) MPN/liter, and 0 to 93 MPN/liter, respectively. Following heavy rainstorms that generated severe flash flooding and heavy discharge of freshwater, salinity decreased, reaching 2.2 to 16.4‰ within 15 days, depending on the site, with a concomitant increase in Vibrio concentration to ca. 10(4) MPN/liter. The highest concentrations were reached with salinities between 10 and 20‰ for V. parahaemolyticus, 10 and 15‰ for V. vulnificus, and 5 and 12‰ for V. cholerae. Thus, an abrupt decrease in salinity caused by heavy rainfall and major flooding favored growth of human-pathogenic Vibrio spp. and their proliferation in the Languedocian lagoons. Based on these results, it is recommended that temperature and salinity monitoring be done to predict the presence of these Vibrio spp. in shellfish-harvesting areas of the lagoons. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  5. Suspension of oysters reduces the populations of Vibrio parahaemolyticus and Vibrio vulnificus.

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    Cole, K M; Supan, J; Ramirez, A; Johnson, C N

    2015-09-01

    Vibrio parahaemolyticus (Vp) and Vibrio vulnificus (Vv) are associated with the consumption of raw oysters and cause illnesses ranging from simple gastroenteritis to life-threatening septicaemia. These halophilic bacteria are frequently found in marine and estuarine systems, accumulating within the tissues of a number of aquatic organisms and passing on to humans after consumption, through contaminated water, or via open wounds. As benthic organisms capable of filtering 40 gallons of water per hour, sediment is an important source of potentially pathogenic vibrios in oysters destined for raw consumption. This research used off-bottom oyster culture to reduce vibrio concentrations in oysters. Colony hybridization was used to enumerate Vp and Vv in bottom and suspended oysters. Vv and Vp concentrations were generally lower in oysters suspended off-bottom, and suspension decreased vibrio loads in oysters by an average of 13%. Suspension of oysters reduced vibrio concentrations. This study found that oyster suspension significantly reduced some populations of potentially pathogenic vibrios. These results indicate that oyster suspension could be a viable approach for preharvest treatment to reduce illness in consumers of raw oysters. © 2015 The Society for Applied Microbiology.

  6. Characterization of the Hemorrhagic Reaction Caused by Vibrio vulnificus Metalloprotease, a Member of the Thermolysin Family

    OpenAIRE

    Miyoshi, Shin-ichi; Nakazawa, Hiromi; Kawata, Koji; Tomochika, Ken-ichi; Tobe, Kazuo; Shinoda, Sumio

    1998-01-01

    Vibrio vulnificus is an opportunistic human pathogen causing wound infections and septicemia, characterized by hemorrhagic and edematous damage to the skin. This human pathogen secretes a metalloprotease (V. vulnificus protease [VVP]) as an important virulence determinant. When several bacterial metalloproteases including VVP were injected intradermally into dorsal skin, VVP showed the greatest hemorrhagic activity. The level of the in vivo hemorrhagic activity of the bacterial metalloproteas...

  7. Septic arthritis and subsequent fatal septic shock caused by Vibrio vulnificus infection

    DEFF Research Database (Denmark)

    Emamifar, Amir; Asmussen Andreasen, Rikke; Andersen, Nanna Skaarup

    2015-01-01

    Vibrio vulnificus is a rare but potential fatal bacterium that can cause severe infections. Wound infections, primary sepsis and gastroenteritis are the most common clinical features. Septic arthritis caused by V. vulnificus is an atypical presentation that has been reported in only two case...... reports; however, it has not been previously noted in Denmark. The authors report a case of septic arthritis caused by V. vulnificus in an immunocompromised patient. The disease progressed to severe sepsis and subsequent death within 10 h of admission....

  8. Survey on antimicrobial resistance patterns in Vibrio vulnificus and Vibrio cholerae non-O1/non-O139 in Germany reveals carbapenemase-producing Vibrio cholerae in coastal waters

    Directory of Open Access Journals (Sweden)

    Nadja eBier

    2015-10-01

    Full Text Available An increase in the occurrence of potentially pathogenic Vibrio species is expected for waters in Northern Europe as a consequence of global warming. In this context, a higher incidence of Vibrio infections is predicted for the future and forecasts suggest that people visiting and living at the Baltic Sea are at particular risk.This study aimed to investigate antimicrobial resistance patterns among Vibrio vulnificus and Vibrio cholerae non-O1/non-O139 isolates that could pose a public health risk. Antimicrobial susceptibility of 141 V. vulnificus and 184 V. cholerae non-O1/non-O139 strains isolated from German coastal waters (Baltic Sea and North Sea as well as from patients and retail seafood was assessed by broth microdilution and disk diffusion. Both species were susceptible to most of the agents tested (12 subclasses and no multidrug-resistance was observed. Among V. vulnificus isolates, non-susceptibility was exclusively found towards aminoglycosides. In case of V. cholerae, a noticeable proportion of strains was non-susceptible to aminopenicillins and aminoglycosides. In addition, resistance towards carbapenems, quinolones, and folate pathway inhibitors was sporadically observed. Biochemical testing indicated the production of carbapenemases with unusual substrate specificity in four environmental V. cholerae strains. Most antimicrobial agents recommended for treatment of V. vulnificus and V. cholerae non-O1/non-O139 infections were found to be effective in vitro. However, the occurrence of putative carbapenemase producing V. cholerae in German coastal waters is of concern and highlights the need for systematic monitoring of antimicrobial susceptibility in potentially pathogenic Vibrio spp. in Europe.

  9. Effects of ambient exposure, refrigeration, and icing on Vibrio vulnificus and Vibrio parahaemolyticus abundances in oysters.

    Science.gov (United States)

    Jones, J L; Lydon, K A; Kinsey, T P; Friedman, B; Curtis, M; Schuster, R; Bowers, J C

    2017-07-17

    Vibrio vulnificus (Vv) and V. parahaemolyticus (Vp) illnesses are typically acquired through the consumption of raw molluscan shellfish, particularly oysters. As Vibrio spp. are naturally-occurring bacteria, one means of mitigation of illness is achieved by limiting post-harvest growth. In this study, effects of ambient air storage, refrigeration, and icing of oysters on Vibrio spp. abundances were examined at two sites in Alabama (AL) [Dog River (DR) and Cedar Point (CP)] and one site in Delaware Bay, New Jersey (NJ). As the United States shellfish program recommendations include testing for total these organisms and gene targets, Vv and total (tlh) and pathogenic (tdh+ and trh+) Vp were enumerated from samples using MPN-real-time-PCR approaches. Mean Vv and Vp abundances in oysters from AL-DR were lowest in immediately iced samples (2.3 and -0.1 log MPN/g, respectively) and highest in the 5h ambient then refrigerated samples (3.4 and 0.5 log MPN/g, respectively). Similarly, in AL-CP Vv and Vp mean levels in oysters were lowest in immediately iced samples (3.6 and 1.2 log MPN/g, respectively) and highest in 5h ambient then refrigerated samples (5.1 and 3.2 log MPN/g, respectively). Mean levels of pathogenic Vp from AL sites were frequently below the limit of detection (ice resulted in lower Vibrio spp. levels in oysters, compared to those that were refrigerated post-harvest. These results suggest vibriosis risk can be mitigated by shorter storage times and more rapid cooling of oysters, providing data regulatory authorities can use to evaluate Vibrio spp. control plans. Published by Elsevier B.V.

  10. Genome-wide SNP-genotyping array to study the evolution of the human pathogen Vibrio vulnificus biotype 3.

    Science.gov (United States)

    Raz, Nili; Danin-Poleg, Yael; Hayman, Ryan B; Bar-On, Yudi; Linetsky, Alex; Shmoish, Michael; Sanjuán, Eva; Amaro, Carmen; Walt, David R; Kashi, Yechezkel

    2014-01-01

    Vibrio vulnificus is an aquatic bacterium and an important human pathogen. Strains of V. vulnificus are classified into three different biotypes. The newly emerged biotype 3 has been found to be clonal and restricted to Israel. In the family Vibrionaceae, horizontal gene transfer is the main mechanism responsible for the emergence of new pathogen groups. To better understand the evolution of the bacterium, and in particular to trace the evolution of biotype 3, we performed genome-wide SNP genotyping of 254 clinical and environmental V. vulnificus isolates with worldwide distribution recovered over a 30-year period, representing all phylogeny groups. A custom single-nucleotide polymorphism (SNP) array implemented on the Illumina GoldenGate platform was developed based on 570 SNPs randomly distributed throughout the genome. In general, the genotyping results divided the V. vulnificus species into three main phylogenetic lineages and an additional subgroup, clade B, consisting of environmental and clinical isolates from Israel. Data analysis suggested that 69% of biotype 3 SNPs are similar to SNPs from clade B, indicating that biotype 3 and clade B have a common ancestor. The rest of the biotype 3 SNPs were scattered along the biotype 3 genome, probably representing multiple chromosomal segments that may have been horizontally inserted into the clade B recipient core genome from other phylogroups or bacterial species sharing the same ecological niche. Results emphasize the continuous evolution of V. vulnificus and support the emergence of new pathogenic groups within this species as a recurrent phenomenon. Our findings contribute to a broader understanding of the evolution of this human pathogen.

  11. Genome-Wide SNP-Genotyping Array to Study the Evolution of the Human Pathogen Vibrio vulnificus Biotype 3

    Science.gov (United States)

    Hayman, Ryan B.; Bar-On, Yudi; Linetsky, Alex; Shmoish, Michael; Sanjuán, Eva; Amaro, Carmen; Walt, David R.; Kashi, Yechezkel

    2014-01-01

    Vibrio vulnificus is an aquatic bacterium and an important human pathogen. Strains of V. vulnificus are classified into three different biotypes. The newly emerged biotype 3 has been found to be clonal and restricted to Israel. In the family Vibrionaceae, horizontal gene transfer is the main mechanism responsible for the emergence of new pathogen groups. To better understand the evolution of the bacterium, and in particular to trace the evolution of biotype 3, we performed genome-wide SNP genotyping of 254 clinical and environmental V. vulnificus isolates with worldwide distribution recovered over a 30-year period, representing all phylogeny groups. A custom single-nucleotide polymorphism (SNP) array implemented on the Illumina GoldenGate platform was developed based on 570 SNPs randomly distributed throughout the genome. In general, the genotyping results divided the V. vulnificus species into three main phylogenetic lineages and an additional subgroup, clade B, consisting of environmental and clinical isolates from Israel. Data analysis suggested that 69% of biotype 3 SNPs are similar to SNPs from clade B, indicating that biotype 3 and clade B have a common ancestor. The rest of the biotype 3 SNPs were scattered along the biotype 3 genome, probably representing multiple chromosomal segments that may have been horizontally inserted into the clade B recipient core genome from other phylogroups or bacterial species sharing the same ecological niche. Results emphasize the continuous evolution of V. vulnificus and support the emergence of new pathogenic groups within this species as a recurrent phenomenon. Our findings contribute to a broader understanding of the evolution of this human pathogen. PMID:25526263

  12. Vibrio vulnificus Type 6 Secretion System 1 Contains Anti-Bacterial Properties.

    Directory of Open Access Journals (Sweden)

    Selina R Church

    Full Text Available Vibrio vulnificus is a bacterium responsible for severe gastroenteritis, sepsis and wound infections. Gastroenteritis and sepsis are commonly associated with the consumption of raw oysters, whereas wound infection is often associated with the handling of contaminated fish. Although classical virulence factors of this emerging pathogen are well characterised, there remains a paucity of knowledge regarding the general biology of this species. To investigate the presence of previously unreported virulence factors, we applied whole genome sequencing to a panel of ten V. vulnificus strains with varying virulence potentials. This identified two novel type 6 secretion systems (T6SSs, systems that are known to have a role in bacterial virulence and population dynamics. By utilising a range of molecular techniques and assays we have demonstrated the functionality of one of these T6SSs. Furthermore, we have shown that this system is subject to thermoregulation and is negatively regulated by increasing salinity concentrations. This secretion system was also shown to be involved in the killing of V. vulnificus strains that did not possess this system and a model is proposed as to how this interaction may contribute to population dynamics within V. vulnificus strains. In addition to this intra-species killing, this system also contributes to the killing of inter bacterial species and may have a role in the general composition of Vibrio species in the environment.

  13. Pathogenesis of Infection by Clinical and Environmental Strains of Vibrio vulnificus in Iron-Dextran-Treated Mice

    OpenAIRE

    Starks, Angela M.; Schoeb, Trenton R.; Tamplin, Mark L.; Parveen, Salina; Doyle, Thomas J.; Bomeisl, Philip E.; Escudero, Gloria M.; Gulig, Paul A.

    2000-01-01

    Vibrio vulnificus is an opportunistic pathogen that contaminates oysters harvested from the Gulf of Mexico. In humans with compromising conditions, especially excess levels of iron in plasma and tissues, consumption of contaminated seafood or exposure of wounds to contaminated water can lead to systemic infection and disfiguring skin infection with extremely high mortality. V. vulnificus-associated diseases are noted for the rapid replication of the bacteria in host tissues, with extensive ti...

  14. Modeling and forecasting the distribution of Vibrio vulnificus in Chesapeake Bay

    Energy Technology Data Exchange (ETDEWEB)

    Jacobs, John M.; Rhodes, M.; Brown, C. W.; Hood, Raleigh R.; Leight, A.; Long, Wen; Wood, R.

    2014-11-01

    The aim is to construct statistical models to predict the presence, abundance and potential virulence of Vibrio vulnificus in surface waters. A variety of statistical techniques were used in concert to identify water quality parameters associated with V. vulnificus presence, abundance and virulence markers in the interest of developing strong predictive models for use in regional oceanographic modeling systems. A suite of models are provided to represent the best model fit and alternatives using environmental variables that allow them to be put to immediate use in current ecological forecasting efforts. Conclusions: Environmental parameters such as temperature, salinity and turbidity are capable of accurately predicting abundance and distribution of V. vulnificus in Chesapeake Bay. Forcing these empirical models with output from ocean modeling systems allows for spatially explicit forecasts for up to 48 h in the future. This study uses one of the largest data sets compiled to model Vibrio in an estuary, enhances our understanding of environmental correlates with abundance, distribution and presence of potentially virulent strains and offers a method to forecast these pathogens that may be replicated in other regions.

  15. Modeling and forecasting the distribution of Vibrio vulnificus in Chesapeake Bay.

    Science.gov (United States)

    Jacobs, J M; Rhodes, M; Brown, C W; Hood, R R; Leight, A; Long, W; Wood, R

    2014-11-01

    To construct statistical models to predict the presence, abundance and potential virulence of Vibrio vulnificus in surface waters of Chesapeake Bay for implementation in ecological forecasting systems. We evaluated and applied previously published qPCR assays to water samples (n = 1636) collected from Chesapeake Bay from 2007-2010 in conjunction with State water quality monitoring programmes. A variety of statistical techniques were used in concert to identify water quality parameters associated with V. vulnificus presence, abundance and virulence markers in the interest of developing strong predictive models for use in regional oceanographic modeling systems. A suite of models are provided to represent the best model fit and alternatives using environmental variables that allow them to be put to immediate use in current ecological forecasting efforts. Environmental parameters such as temperature, salinity and turbidity are capable of accurately predicting abundance and distribution of V. vulnificus in Chesapeake Bay. Forcing these empirical models with output from ocean modeling systems allows for spatially explicit forecasts for up to 48 h in the future. This study uses one of the largest data sets compiled to model Vibrio in an estuary, enhances our understanding of environmental correlates with abundance, distribution and presence of potentially virulent strains and offers a method to forecast these pathogens that may be replicated in other regions. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

  16. Abundance and antibiotic susceptibility of Vibrio spp. isolated from microplastics

    Science.gov (United States)

    Laverty, A. L.; Darr, K.; Dobbs, F. C.

    2016-02-01

    In recent years, there has been a growing concern for `microplastics' (particles resistance profiles of Vibrio spp. found on them. We collected 22 microplastic pieces, paired seawater samples, and from them cultured 44 putative Vibrio spp. isolates, 18 of which were PCR-confirmed as V. parahaemolyticus and 3 as V. vulnificus. There were no PCR-confirmed V. cholerae isolates. We used the Kirby-Bauer disk diffusion susceptibility test to examine the isolates' response to six antibiotics: chloramphenicol (30μg), gentamicin (10μg), ampicillin (10μg), streptomycin (10μg), tetracycline (30μg), and rifampin (5μg). Vibrio isolates were susceptible to three or more of the six antibiotics tested and all were susceptible to tetracycline and chloramphenicol. There were no apparent differences between the antibiotic susceptibilities of vibrios isolated from microplastics compared to those from the water column. In every instance tested, vibrios on microplastics were enriched by at least two orders of magnitude compared to those from paired seawater samples. This study demonstrates that microplastic particles serve as a habitat for Vibrio species, in particular V. vulnificus and V. parahaemolyticus, confirming the conjecture of Zettler et al. (2013) that plastics may serve as a vector for these and other potentially pathogenic bacteria.

  17. Vibrio vulnificus produces quorum sensing signals of the AHL-class

    DEFF Research Database (Denmark)

    Valiente, E.; Bruhn, Jesper Bartholin; Nielsen, Kristian Fog

    2009-01-01

    Vibrio vulnificus is an aquatic pathogenic bacterium that can cause vibriosis in humans and fish. The species is subdivided into three biotypes with the fish-virulent strains belonging to biotype 2. The quorum sensing (QS) phenomenon mediated by furanosyl borate diester or autoinducer 2 (AI-2) ha...... biotype 2 strains. No known AHL-related gene was detected by PCR or Southern blot suggesting that AHL-related genes in V. vulnificus are different from those found in other Gram-negative bacteria....... was detected when AHL-positive strains were grown in low-nutrient medium [modified sea water yeast extract (MSWYE)] but not in rich media (tryptic soy broth or brain–heart infusion) and its production was enhanced when blood factors were added to MSWYE. C4-HL was detected in vivo, in eels infected with AHL-positive...

  18. Clinical Usefulness of Real-Time Polymerase Chain Reaction for the Diagnosis of Vibrio vulnificus Infection Using Skin and Soft Tissues.

    Science.gov (United States)

    Lee, Jun-Young; Kim, Seok Won; Kim, Dong-Min; Yun, Na Ra; Kim, Choon-Mee; Lee, Sang-Hong

    2017-08-01

    Vibrio vulnificus is a halophilic gram-negative bacillus isolated in seawater, fish, and shellfish. Infection by V. vulnificus is the most severe food-borne infection reported in the United States of America. Here, we aimed to examine the clinical usefulness of polymerase chain reaction (PCR) using tissue specimens other than blood samples as a diagnostic tool for V. vulnificus infection. A retrospective study was conducted with patients who underwent real-time PCR of toxR in both blood and skin tissues, including serum, bullae, swab, and operation room specimens, between 2006 and 2009. The median V. vulnificus DNA load of 14 patients in real-time PCR analysis of serum at the time of admission was 638.5 copies/mL blood, which was within the interquartile range (IQR: 37-3,225). In contrast, the median value by real-time PCR using the first tissue specimen at the time of admission was 16,650 copies/mL tissue fluid (IQR: 4,419-832,500). This difference was statistically significant (P = 0.022). DNA copy numbers in tissues were less affected by short-term antibiotic administration than that in blood samples, and antibiotic administration increased the DNA copy number in some patients. We found, for the first time, that DNA copy numbers in tissues of patients infected by V. vulnificus were higher than those in blood samples. Additionally, skin lesions were more useful than blood samples as specimens for PCR analysis in patients administered antibiotics for V. vulnificus infection before admission.

  19. Necrotizing soft-tissue infections and sepsis caused by Vibrio vulnificus compared with those caused by Aeromonas species.

    Science.gov (United States)

    Tsai, Yao-Hung; Hsu, Robert Wen-Wei; Huang, Tsung-Jen; Hsu, Wei-Hsiu; Huang, Kuo-Chin; Li, Yen-Yao; Peng, Kuo-Ti

    2007-03-01

    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 outcome. The cases of thirty-two patients with necrotizing soft-tissue infections and sepsis caused by Vibrio vulnificus (seventeen patients) and Aeromonas species (fifteen patients) were retrospectively reviewed over a four-year period. Surgical débridement or immediate limb amputation was initially performed in all patients. Demographic data, underlying diseases, laboratory results, and clinical outcome were analyzed for each patient in both groups. Six patients in the Vibrio vulnificus group and four patients in the Aeromonas group died. The patients who died had significantly lower serum albumin levels than did the patients who survived (p tissue infection with Vibrio vulnificus (contact with seawater or raw seafood) or Aeromonas species (contact with fresh or brackish water, soil, or wood). Early fasciotomy and culture-directed antimicrobial therapy should be aggressively performed in those patients with hypotensive shock, leukopenia, severe hypoalbuminemia, and underlying chronic illness, especially a combination of hepatic dysfunction and diabetes mellitus.

  20. Portrait of a viral infection: The infection cycle of Vibrio vulnificus phage VvAW1 visualized through plaque assay, electron microscopy, and proteomics

    Science.gov (United States)

    Clah, K. E. Y.; Nigro, O. D.; Miranda, J.; Schvarcz, C.; Culley, A.; Saito, M. A.; Steward, G.

    2016-02-01

    The bacterium Vibrio vulnificus is an opportunistic human pathogen that thrives in warm brackish waters. Viral infection is one of several mechanisms influencing the population dynamics of this bacterium in the natural environment. V. vulnificus-specific viruses have been isolated; however, the details of their infection cycle have not been reported. As a result, our current understanding of the interaction between the bacterium and its viruses in the environment is limited. To better understand the infection process, a strain of V. vulnificus (V93D1V) and its bacteriophage, Vibrio phage VvAW1, were isolated from the estuarine waters of the Ala Wai Canal, HI. A time-series infection experiment was conducted with the virus-host pair in which samples were collected every ten minutes for eighty minutes post-infection for analysis by plaque assay, electron microscopy, and proteomics. Using electron microscopy, visibly infected bacteria were observed forty minutes after the introduction of the virus, signaling the end of the eclipse period. The peak of infection occurred at seventy minutes with an average viral load of 78 viruses per bacterium. The percentage of visibly infected bacteria reached a maximum just prior to a rise in free viruses in the culture, indicating the end of the latent period. The percentage of infected cells that lysed was low and there was little effect on the bacterial population growth rate. Analysis of the proteome revealed that protein expression patterns, in particular capsid and other structural proteins, closely follow the timing of the observed infection cycle. Together, these analyses provided the first detailed view of a viral infection in a highly lethal aquatic bacterium. The apparent temperate nature of this virus suggests that it can be a source of mortality to V. vulnificus, but has evolved to avoid total destruction of its host by complete lysis, a characteristic that helps ensure its replication in subsequent generations.

  1. Effectiveness of icing as a postharvest treatment for control of Vibrio vulnificus and Vibrio parahaemolyticus in the eastern oyster (Crassostrea virginica).

    Science.gov (United States)

    Melody, Kevin; Senevirathne, Reshani; Janes, Marlene; Jaykus, Lee Ann; Supan, John

    2008-07-01

    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 Vibrio spp. during wholesale and retail handling for 2 weeks postharvest were also monitored. On-board icing achieved temperature reductions in all sacks in accordance with the National Shellfish Sanitation Program standard, but dockside icing did not meet this standard. Based on one-way analysis of variance, the only statistically significant relationship between Vibrio levels and treatment occurred for samples harvested in August; in this case, the levels of V. vulnificus in the noniced oysters were significantly higher (P iced on-board. When analyzing counts over the 14-day storage period, using factorial analysis, there were statistically significant differences in V. vulnificus and V. parahaemolyticus levels by sample date and/or treatment (P iced) oysters had significantly higher gaping (approximately 20%) after 1 week in cold storage than did noniced oysters (approximately 10%) and gaping increased significantly by day 14 of commercial storage. On-board and dockside icing did not predictably reduce the levels of V. vulnificus or V. parahaemolyticus in oysters, and icing negatively impacted oyster survival during subsequent cold storage.

  2. Uncertainty in model predictions of Vibrio vulnificus response to climate variability and change: a Chesapeake Bay case study.

    Directory of Open Access Journals (Sweden)

    Erin A Urquhart

    Full Text Available The effect that climate change and variability will have on waterborne bacteria is a topic of increasing concern for coastal ecosystems, including the Chesapeake Bay. Surface water temperature trends in the Bay indicate a warming pattern of roughly 0.3-0.4°C per decade over the past 30 years. It is unclear what impact future warming will have on pathogens currently found in the Bay, including Vibrio spp. Using historical environmental data, combined with three different statistical models of Vibrio vulnificus probability, we explore the relationship between environmental change and predicted Vibrio vulnificus presence in the upper Chesapeake Bay. We find that the predicted response of V. vulnificus probability to high temperatures in the Bay differs systematically between models of differing structure. As existing publicly available datasets are inadequate to determine which model structure is most appropriate, the impact of climatic change on the probability of V. vulnificus presence in the Chesapeake Bay remains uncertain. This result points to the challenge of characterizing climate sensitivity of ecological systems in which data are sparse and only statistical models of ecological sensitivity exist.

  3. Uncertainty in Model Predictions of Vibrio Vulnificus Response to Climate Variability and Change: A Chesapeake Bay Case Study

    Science.gov (United States)

    Urquhart, Erin A.; Zaitchik, Benjamin F.; Waugh, Darryn W.; Guikema, Seth D.; Del Castillo, Carlos E.

    2014-01-01

    The effect that climate change and variability will have on waterborne bacteria is a topic of increasing concern for coastal ecosystems, including the Chesapeake Bay. Surface water temperature trends in the Bay indicate a warming pattern of roughly 0.3-0.4 C per decade over the past 30 years. It is unclear what impact future warming will have on pathogens currently found in the Bay, including Vibrio spp. Using historical environmental data, combined with three different statistical models of Vibrio vulnificus probability, we explore the relationship between environmental change and predicted Vibrio vulnificus presence in the upper Chesapeake Bay. We find that the predicted response of V. vulnificus probability to high temperatures in the Bay differs systematically between models of differing structure. As existing publicly available datasets are inadequate to determine which model structure is most appropriate, the impact of climatic change on the probability of V. vulnificus presence in the Chesapeake Bay remains uncertain. This result points to the challenge of characterizing climate sensitivity of ecological systems in which data are sparse and only statistical models of ecological sensitivity exist.

  4. Impact of Hurricane Irene on Vibrio vulnificus and Vibrio parahaemolyticus concentrations in surface water, sediment, and cultured oysters in the Chesapeake Bay, MD, USA.

    Science.gov (United States)

    Shaw, Kristi S; Jacobs, John M; Crump, Byron C

    2014-01-01

    To determine if a storm event (i.e., high winds, large volumes of precipitation) could alter concentrations of Vibrio vulnificus and V. parahaemolyticus in aquacultured oysters (Crassostrea virginica) and associated surface water and sediment, this study followed a sampling timeline before and after Hurricane Irene impacted the Chesapeake Bay estuary in late August 2011. Aquacultured oysters were sampled from two levels in the water column: surface (0.3 m) and near-bottom (just above the sediment). Concentrations of each Vibrio spp. and associated virulence genes were measured in oysters with a combination of real-time PCR and most probable number (MPN) enrichment methods, and in sediment and surface water with real-time PCR. While concentration shifts of each Vibrio species were apparent post-storm, statistical tests indicated no significant change in concentration for either Vibrio species by location (surface or near bottom oysters) or date sampled (oyster tissue, surface water, and sediment concentrations). V. vulnificus in oyster tissue was correlated with total suspended solids (r = 0.41, P = 0.04), and V. vulnificus in sediment was correlated with secchi depth (r = -0.93, P depth [r = -0.48, P = 0.02 (sediment); r = -0.97, P <0.01 (surface water)] and tidal height [r = -0.96, P <0.01 (sediment), r = -0.59, P <0.01 (surface water)]. The concentrations of Vibrio spp. were higher in oysters relative to other studies (average V. vulnificus 4 × 10(5) MPN g(-1), V. parahaemolyticus 1 × 10(5) MPN g(-1)), and virulence-associated genes were detected in most oyster samples. This study provides a first estimate of storm-related Vibrio density changes in oyster tissues, sediment, and surface water at an aquaculture facility in the Chesapeake Bay.

  5. Vibrio vulnificus MO6-24/O Lipopolysaccharide Stimulates Superoxide Anion, Thromboxane B2, Matrix Metalloproteinase-9, Cytokine and Chemokine Release by Rat Brain Microglia in Vitro

    Directory of Open Access Journals (Sweden)

    Alejandro M. S. Mayer

    2014-03-01

    Full Text Available Although human exposure to Gram-negative Vibrio vulnificus (V. vulnificus lipopolysaccharide (LPS has been reported to result in septic shock, its impact on the central nervous system’s innate immunity remains undetermined. The purpose of this study was to determine whether V. vulnificus MO6-24/O LPS might activate rat microglia in vitro and stimulate the release of superoxide anion (O2−, a reactive oxygen species known to cause oxidative stress and neuronal injury in vivo. Brain microglia were isolated from neonatal rats, and then treated with either V. vulnificus MO6-24/O LPS or Escherichia coli O26:B6 LPS for 17 hours in vitro. O2− was determined by cytochrome C reduction, and matrix metalloproteinase-2 (MMP-2 and MMP-9 by gelatinase zymography. Generation of cytokines tumor necrosis factor alpha (TNF-α, interleukin-1 alpha (IL-1α, IL-6, and transforming growth factor-beta 1 (TGF-β1, chemokines macrophage inflammatory protein (MIP-1α/chemokine (C-C motif ligand 3 (CCL3, MIP-2/chemokine (C-X-C motif ligand 2 (CXCL2, monocyte chemotactic protein-1 (MCP-1/CCL2, and cytokine-induced neutrophil chemoattractant-2alpha/beta (CINC-2α/β/CXCL3, and brain-derived neurotrophic factor (BDNF, were determined by specific immunoassays. Priming of rat microglia by V. vulnificus MO6-24/O LPS in vitro yielded a bell-shaped dose-response curve for PMA (phorbol 12-myristate 13-acetate-stimulated O2− generation: (1 0.1–1 ng/mL V. vulnificus LPS enhanced O2− generation significantly but with limited inflammatory mediator generation; (2 10–100 ng/mL V. vulnificus LPS maximized O2− generation with concomitant release of thromboxane B2 (TXB2, matrix metalloproteinase-9 (MMP-9, and several cytokines and chemokines; (3 1000–100,000 ng/mL V. vulnificus LPS, with the exception of TXB2, yielded both attenuated O2− production, and a progressive decrease in MMP-9, cytokines and chemokines investigated. Thus concentration-dependent treatment of

  6. A comparative epizootiologic study of the two fish-pathogenic serovars of Vibrio vulnificus biotype 2.

    Science.gov (United States)

    Fouz, B; Llorens, A; Valiente, E; Amaro, C

    2010-05-01

    Vibrio vulnificus biotype 2 is subdivided into two main serovars, serovar E, able to infect fish and humans, and serovar A, only virulent for fish. Serovar E emerged in 1976 as the causative agent of a haemorrhagic septicaemia (warm-water vibriosis) affecting eels cultured in brackish water. Serovar A emerged in 2000 in freshwater-cultured eels vaccinated against serovar E, causing warm-water vibriosis with fish showing a haemorrhagic intestine as the main differential sign. The aim of the present work was to compare the disease caused by both serovars in terms of transmission routes, portals of entry and host range. Results of bath, patch-contact and oral-anal challenges demonstrated that both serovars spread through water and infect healthy eels, serovar A entering mainly by the anus and serovar E by the gills. The course of the disease under laboratory conditions was similar for both serovars in terms of transmission and dependence of degree of virulence on water parameters (temperature and salinity). However, the decrease in degree of virulence in fresh water was significantly greater in serovar E than in serovar A. Finally, both serovars proved pathogenic for tilapia, sea bass and rainbow trout, but not for sea bream, with significant differences in degree of virulence only in rainbow trout. In conclusion, serovar A seems to represent a new antigenic form of V. vulnificus biotype 2 with an unusual portal of entry and is better adapted to fresh water than serovar E.

  7. Variable Virulence of Biotype 3 Vibrio vulnificus due to MARTX Toxin Effector Domain Composition.

    Science.gov (United States)

    Kim, Byoung Sik; Gavin, Hannah E; Satchell, Karla J F

    2017-01-01

    Vibrio vulnificus is an environmental organism that causes septic human infections characterized by high morbidity and mortality. The annual incidence and global distribution of this pathogen are increasing as ocean waters warm. Clinical strains exhibit variations in the primary virulence toxin, suggesting a potential for the emergence of new strains with altered virulence properties. A clonal outbreak of tilapia-associated wound infections in Israel serves as a natural experiment for the sudden emergence of a new V. vulnificus strain. The effector domain content of the multifunctional autoprocessing RTX (MARTX) toxin of the outbreak-associated biotype 3 (BT3) strains was previously shown to harbor a modification generated by recombination. The modification introduced an actin-induced adenylate cyclase effector domain (ExoY) and an effector domain that disrupts the Golgi organelle (DmX). Here, we report that the exchange of these effector domains for a putative progenitor biotype 1 toxin arrangement produces a toxin that slows the lysis kinetics of targeted epithelial cells but increases cellular rounding phenotypes in response to bacteria. In addition, replacing the biotype 3 toxin variant with the putative progenitor biotype 1 variant renders the resulting strain significantly more virulent in mice. This suggests that the exchange of MARTX effector domains during the emergence of BT3 generated a toxin with reduced toxin potency, resulting in decreased virulence of this outbreak-associated strain. We posit that selection for reduced virulence may serve as a route for this lethal infectious agent to enter the human food chain by allowing it to persist in natural hosts. IMPORTANCEVibrio vulnificus is a serious infection linked to climate change. The virulence capacity of these bacteria can vary by gene exchange, resulting in new variants of the primary virulence toxin. In this study, we tested whether the emergence of an epidemic strain of V. vulnificus with a

  8. Prevalence study of Vibrio species and frequency of the virulence genes of Vibrio parahaemolyticus isolated from fresh and salted shrimps in Genaveh seaport

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    S Hosseini

    2014-08-01

    Full Text Available Vibrio species are important seafood-borne pathogens that are responsible for 50-70% of gasteroenteritis. The present study was carried out in order to determine the prevalence of Vibrio species and the distribution of tdh, tlh and trh virulence genes in Vibrio parahaemolyticus isolated from fresh and salted shrimp samples. Totally, 60 fresh and salted shrimp samples were collected from the Genaveh seaport. Microbial culture was used to isolate Vibrio species. In addition, the presences of Vibrio parahaemolyticus, Vibrio cholera, Vibrio vulnificus and Vibrio harveyi and the virulence genes of V. parahaemolyticus were studied using the PCR method. Results showed that 20% of fresh and 23.33% of salted shrimp samples were positive for Vibrio species. In studied samples, V. vulnificus had the highest prevalence rate (8.33%, while V. cholera had the lowest prevalence rate (1.66%. From a total of 4 detected V. parahaemolyticus, all of them had tlh gene (100%. The distribution of tdh and trh genes in isolated V. parahaemolyticus strains were 50% and 25%, respectively. High prevalence of Vibrio species and especially virulent V. parahaemolyticus in samples confirmed the lack of hygienic condition in the production and distribution centers of shrimp.

  9. Impact of Hurricane Irene on Vibrio vulnificus and Vibrio parahaemolyticus concentrations in surface water, sediment and cultured oysters in the Chesapeake Bay, Maryland, USA

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    Kristi S Shaw

    2014-05-01

    Full Text Available To determine if a storm event (i.e., high winds, large volumes of precipitation could alter concentrations of Vibrio vulnificus and Vibrio parahaemolyticus in aquacultured oysters (Crassostrea virginica and associated surface water and sediment, this study followed a sampling timeline before and after Hurricane Irene impacted the Chesapeake Bay estuary in late August 2011. Aquacultured oysters were sampled from two levels in the water column: surface 0.3 m and near-bottom just above the sediment. Concentrations of each Vibrio spp. and associated virulence genes were measured in oysters with a combination of real-time PCR and most probable number enrichment methods, and in sediment and surface water with real-time PCR. While concentration shifts of each Vibrio species were apparent post-storm, statistical tests indicated no significant change in concentration change for either Vibrio species by location (surface or near bottom oysters or date sampled (oyster tissue, surface water and sediment concentrations. V. vulnificus in oyster tissue was correlated with total suspended solids (r=0.41, p=0.04, and V. vulnificus in sediment was correlated with secchi depth (r=-0.93, p< 0.01, salinity (r=-0.46, p=0.02, tidal height (r=-0.45, p=0.03, and surface water V. vulnificus (r=0.98, p< 0.01. V. parahaemolyticus in oyster tissue did not correlate with environmental measurements, but V. parahaemolyticus in sediment and surface water correlated with several measurements including secchi depth (r=-0.48, p=0.02[sediment]; r=-0.97 p< 0.01[surface water] and tidal height (r=-0.96. p< 0.01[sediment], r=-0.59,p< 0.01 [surface water]. The concentrations of Vibrio spp. were higher in oysters relative to other studies (average V. vulnificus 4x105 MPN g-1, V. parahaemolyticus 1x105 MPN g-1, and virulence-associated genes were detected in most oyster samples. This study provides a first estimate of storm-related Vibrio density changes in oyster tissues, sediment and

  10. Necrotising fasciitis causing compartment syndrome of the forearm and septic shock due to Vibrio vulnificus: a case report.

    Science.gov (United States)

    Leechavengvongs, Somsak; Jidpugdeebodin, Suwanee; Milindankura, Samaniya

    2006-01-01

    Compartment syndrome caused by necrotising fasciitis has rarely been described. We report a case of systemic Vibrio vulnificus necrotising fasciitis presented with compartmental syndrome of the forearm and septic shock. The patient was treated with systemic antibiotic treatment and urgent surgical decompression followed by multiple necrotic tissue debridements. The patient recovered with some limited motion of the hand function. Prompt recognition and immediate treatment with antibiotics and surgical intervention are essential.

  11. Temperature effect on high salinity depuration of Vibrio vulnificus and V. parahaemolyticus from the Eastern oyster (Crassostrea virginica).

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    Larsen, A M; Rikard, F S; Walton, W C; Arias, C R

    2015-01-02

    Vibrio vulnificus (Vv) and Vibrio parahaemolyticus (Vp) are opportunistic human pathogens naturally associated with the Eastern oyster Crassostrea virginica. The abundances of both pathogens in oysters are positively correlated with temperature, thus ingestion of raw oysters during the warm summer months is a risk factor for contracting illness from these bacteria. Current post-harvest processing (PHP) methods for elimination of these pathogens are expensive and kill the oyster, changing their organoleptic properties and making them less appealing to some consumers. High salinity has proven effective in reducing Vv numbers in the wild and our research aims at developing an indoor recirculating system to reduce pathogenic Vibrios while maintaining the taste and texture of live oysters. The goal of this study was to determine the influence of temperature on the efficacy of high salinity depuration. Vv was enumerated as most probable number (MPN) per gram of oyster tissue using the FDA-approved modified cellobiose polymyxin colistin (mCPC) protocol and with an alternative Vibrio specific media CHROMagar™ Vibrio (CaV). CaV was also used to quantify Vp. Oysters were held at 35 psu for 10 days at three temperatures: low (20°C), mid (22.5°C) and high (25°C). There was no difference in MPN/g of Vv between media; however more Vv isolates were obtained from mCPC than CaV. There was no significant effect of temperature on reduction of Vv or Vp throughout depuration but there was a tendency for low temperatures to be less effective than the higher ones. High salinity resulted in a significant decrease in Vv by day 3 and again by day 10, and a decrease in Vp by day 3. Oyster condition indices were maintained throughout depuration and mortality was low (4% across three trials). Overall these results support the use of mCPC for Vv enumeration and demonstrate the promise of high salinity depuration for PHP of the Eastern oyster. The trend for lower temperatures to be less

  12. Correlations between Clinical Features and Mortality in Patients with Vibrio vulnificus Infection.

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    Hong Zhao

    Full Text Available Vibrio vulnificus is a common gram-negative bacterium, which might cause morbidity and mortality in patients following consumption of seafood or exposure to seawater in Southeast China. We retrospectively analyzed clinical data of patients with laboratory confirmed V. vulnificus infection. Twenty one patients were divided into a survival group and a non-surviving (or death group according to their clinical outcome. Clinical data and measurements were statistically analyzed. Four patients (19.05% died and five patients gave positive cultures from bile fluid, and 16 other patients gave positive culture from blood or blisters. Ten patients (47.62% had an underlying liver disease and marine-related events were found in sixteen patients (76.2%. Patients with heavy drinking habits might be at increased mortality (p = 0.028. Clinical manifestations of cellulitis (47.6%, septic shock (42.9% and multiple organ failure (28.6% were statistically significant when comparing survivors and non-survivors (p = 0.035, p = 0.021 and p = 0.003, respectively. The laboratory results, including hemoglobin < 9.0 g/L (p = 0.012, platelets < 2.0 × 109 /L, prothrombin time activity (PTA <20%, decreased serum creatinine and increased urea nitrogen were statistically significant (p = 0.012, p = 0.003, p = 0.028 and p = 0.028, respectively. Patients may be at a higher risk of mortality under situations where they have a history of habitual heavy alcoholic drink consumption (p = 0.028, OR = 22.5, 95%CI 1.5-335.3, accompanied with cellulitis, shock, multiple organ failure, and laboratory examinations that are complicated by decreased platelets, hemoglobin and significantly prolonged prothrombin time (PT.

  13. MARTX Toxin in the Zoonotic Serovar of Vibrio vulnificus Triggers an Early Cytokine Storm in Mice

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    Celia Murciano

    2017-07-01

    Full Text Available Vibrio vulnificus biotype 2-serovar E is a zoonotic clonal complex that can cause death by sepsis in humans and fish. Unlike other biotypes, Bt2 produces a unique type of MARTXVv (Multifunctional-Autoprocessive-Repeats-in-Toxin; RtxA13, which is encoded by a gene duplicated in the pVvBt2 plasmid and chromosome II. In this work, we analyzed the activity of this toxin and its role in human sepsis by performing in vitro, ex vivo, and in vivo assays. First, we demonstrated that the ACD domain, present exclusively in this toxin variant, effectively has an actin-cross-linking activity. Second, we determined that the whole toxin caused death of human endotheliocytes and monocytes by lysis and apoptosis, respectively. Finally, we tested the hypothesis that RtxA13 contributes to human death caused by this zoonotic serovar by triggering an early cytokine storm in blood. To this end, we used a Bt2-SerE strain (R99 together with its rtxA13 deficient mutant, and a Bt1 strain (YJ016 producing RtxA11 (the most studied MARTXVv together with its rtxA11 deficient mutant, as controls. Our results showed that RtxA13 was essential for virulence, as R99ΔΔrtxA13 was completely avirulent in our murine model of infection, and that R99, but not strain YJ016, induced an early, strong and dysregulated immune response involving the up-regulation of a high number of genes. This dysregulated immune response was directly linked to RtxA13. Based on these results and those obtained ex vivo (human blood, we propose a model of infection for the zoonotic serovar of V. vulnificus, in which RtxA13 would act as a sepsis-inducing toxin.

  14. The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus.

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    Jin Hwan Park

    2015-09-01

    Full Text Available A transcriptome analysis identified Vibrio vulnificus cabABC genes which were preferentially expressed in biofilms. The cabABC genes were transcribed as a single operon. The cabA gene was induced by elevated 3',5'-cyclic diguanylic acid (c-di-GMP and encoded a calcium-binding protein CabA. Comparison of the biofilms produced by the cabA mutant and its parent strain JN111 in microtiter plates using crystal-violet staining demonstrated that CabA contributed to biofilm formation in a calcium-dependent manner under elevated c-di-GMP conditions. Genetic and biochemical analyses revealed that CabA was secreted to the cell exterior through functional CabB and CabC, distributed throughout the biofilm matrix, and produced as the biofilm matured. These results, together with the observation that CabA also contributes to the development of rugose colony morphology, indicated that CabA is a matrix-associated protein required for maturation, rather than adhesion involved in the initial attachment, of biofilms. Microscopic comparison of the structure of biofilms produced by JN111 and the cabA mutant demonstrated that CabA is an extracellular matrix component essential for the development of the mature biofilm structures in flow cells and on oyster shells. Exogenously providing purified CabA restored the biofilm- and rugose colony-forming abilities of the cabA mutant when calcium was available. Circular dichroism and size exclusion analyses revealed that calcium binding induces CabA conformational changes which may lead to multimerization. Extracellular complementation experiments revealed that CabA can assemble a functional matrix only when exopolysaccharides coexist. Consequently, the combined results suggested that CabA is a structural protein of the extracellular matrix and multimerizes to a conformation functional in building robust biofilms, which may render V. vulnificus to survive in hostile environments and reach a concentrated infective dose.

  15. The cabABC Operon Essential for Biofilm and Rugose Colony Development in Vibrio vulnificus

    Science.gov (United States)

    Park, Jin Hwan; Jo, Youmi; Jang, Song Yee; Kwon, Haenaem; Irie, Yasuhiko; Parsek, Matthew R.; Kim, Myung Hee; Choi, Sang Ho

    2015-01-01

    A transcriptome analysis identified Vibrio vulnificus cabABC genes which were preferentially expressed in biofilms. The cabABC genes were transcribed as a single operon. The cabA gene was induced by elevated 3′,5′-cyclic diguanylic acid (c-di-GMP) and encoded a calcium-binding protein CabA. Comparison of the biofilms produced by the cabA mutant and its parent strain JN111 in microtiter plates using crystal-violet staining demonstrated that CabA contributed to biofilm formation in a calcium-dependent manner under elevated c-di-GMP conditions. Genetic and biochemical analyses revealed that CabA was secreted to the cell exterior through functional CabB and CabC, distributed throughout the biofilm matrix, and produced as the biofilm matured. These results, together with the observation that CabA also contributes to the development of rugose colony morphology, indicated that CabA is a matrix-associated protein required for maturation, rather than adhesion involved in the initial attachment, of biofilms. Microscopic comparison of the structure of biofilms produced by JN111 and the cabA mutant demonstrated that CabA is an extracellular matrix component essential for the development of the mature biofilm structures in flow cells and on oyster shells. Exogenously providing purified CabA restored the biofilm- and rugose colony-forming abilities of the cabA mutant when calcium was available. Circular dichroism and size exclusion analyses revealed that calcium binding induces CabA conformational changes which may lead to multimerization. Extracellular complementation experiments revealed that CabA can assemble a functional matrix only when exopolysaccharides coexist. Consequently, the combined results suggested that CabA is a structural protein of the extracellular matrix and multimerizes to a conformation functional in building robust biofilms, which may render V. vulnificus to survive in hostile environments and reach a concentrated infective dose. PMID:26406498

  16. Prevalence of Vibrio parahaemolyticus, and Vibrio vulnificus in blue crabs (Callinectes sapidus), seawater and sediments of the Maryland Coastal Bays.

    Science.gov (United States)

    Rodgers, C; Parveen, S; Chigbu, P; Jacobs, J; Rhodes, M; Harter-Dennis, J

    2014-10-01

    To determine the prevalence of total and pathogenic Vibrio parahaemolyticus (Vp) and V. vulnificus (Vv) in blue crabs, water and sediment from the Maryland Coastal Bays (MCBs), USA. Crab, haemolymph, sediment and seawater samples were collected monthly from four sites in MCBs from February 2012 through October 2012 with environmental parameters recorded. The most-probable-number (MPN) methodology was used to enumerate Vp and Vv with presumptive colonies and the presence of virulence markers confirmed using polymerase chain reaction (PCR). Results indicate that blue crabs contained both Vp and Vv at densities (7·28 and 5·43 log MPN g(-1) , respectively) higher than those reported for bivalves. In addition, markers for clinically relevant strains of both species were detected in over 30% of samples. Haemolymph, sediment and seawater samples were also routinely positive for both species and clinically relevant strains, but generally at lower densities than found in crabs (4·27, 3·28, and 2·39 log MPN g(-1) per ml(-1) Vp, and 4·28, 2·49 and 2·38 log MPN g(-1) per ml(-1) Vv). Blue crabs concentrate Vp and Vv at levels greater than found in water or sediment. While changes in abundance associated with temperature are apparent, there is little evidence to support differences among sampling locations. These results highlight the potential for blue crab related vibriosis and the importance of proper handling, cooking and care of this popular seafood before consumption. © 2014 The Society for Applied Microbiology.

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

    Science.gov (United States)

    Griffitt, Kimberly J; Grimes, D Jay

    2013-04-01

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

  18. Vibrio vulnificus biotype 3 multifunctional autoprocessing RTX toxin is an adenylate cyclase toxin essential for virulence in mice.

    Science.gov (United States)

    Ziolo, Kevin J; Jeong, Hee-Gon; Kwak, Jayme S; Yang, Shuangni; Lavker, Robert M; Satchell, Karla J F

    2014-05-01

    Vibrio vulnificus is an environmental organism that causes both food-borne and wound infections with high morbidity and mortality in humans. The annual incidence and global distribution of infections associated with this pathogen are increasing with climate change. In the late 1990s, an outbreak of tilapia-associated wound infections in Israel was linked to a previously unrecognized variant of V. vulnificus designated biotype 3. The sudden emergence and clonality of the outbreak suggest that this strain may be a true newly emergent pathogen with novel virulence properties compared to those of other V. vulnificus strains. In a subcutaneous infection model to mimic wound infection, the multifunctional autoprocessing RTX (MARTX) toxin of biotype 3 strains was shown to be an essential virulence factor contributing to highly inflammatory skin wounds with severe damage affecting every tissue layer. We conducted a sequencing-based analysis of the MARTX toxin and found that biotype 3 MARTX toxin has an effector domain structure distinct from that of either biotype 1 or biotype 2. Of the two new domains identified, a domain similar to Pseudomonas aeruginosa ExoY was shown to confer adenylate cyclase activity on the MARTX toxin. This is the first demonstration that the biotype 3 MARTX toxin is essential for virulence and that the ExoY-like MARTX effector domain is a catalytically active adenylate cyclase.

  19. Role of extracellular matrix protein CabA in resistance of Vibrio vulnificus biofilms to decontamination strategies.

    Science.gov (United States)

    Park, Jin Hwan; Lee, Byungho; Jo, Youmi; Choi, Sang Ho

    2016-11-07

    Biofilms are recalcitrant and raise safety problems in the food industry. In this study, the role of CabA, an extracellular matrix protein, in the resistance of the biofilms of Vibrio vulnificus, a foodborne pathogen, to decontamination strategies was investigated. Biofilms of the cabA mutant revealed reduced resistance to detachment by vibration and disinfection by sodium hypochlorite compared to the biofilms of the parental wild type in vitro. The reduced resistance of the cabA mutant biofilms was complemented by introducing a recombinant cabA, indicating that the reduced resistance of the cabA mutant biofilms is caused by the inactivation of cabA. The expression of cabA was induced in cells bound to oyster, the primary vehicle of the pathogen. The cabA mutant biofilms on oyster are defective in biomass and resistance to detachment and disinfection. The bacterial cells in the wild-type biofilms are clustered by filaments which are not apparent in the cabA mutant biofilms. The combined results indicated that CabA contributes to the structural integrity of V. vulnificus biofilms possibly by forming filaments in the matrix and thus rendering the biofilms robust, suggesting that CabA could be a target to control V. vulnificus biofilms on oyster. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Light scattering sensor for real-time identification of Vibrio parahaemolyticus, V. vulnificus and V. cholera colonies on solid agar plates

    Science.gov (United States)

    The three most common pathogenic species of Vibrio, V. cholerae, V. parahemolyticus and V. vulnificus, are of major concern as water- and food-borne pathogens because of an increasing incidence of water and seafood related outbreaks and illnesses worldwide. Current methods are time-consuming and req...

  1. Zebrafish fed on recombinant Artemia expressing epinecidin-1 exhibit increased survival and altered expression of immunomodulatory genes upon Vibrio vulnificus infection.

    Science.gov (United States)

    Jheng, Yu-Hsuan; Lee, Lin-Han; Ting, Chen-Hung; Pan, Chieh-Yu; Hui, Cho-Fat; Chen, Jyh-Yih

    2015-01-01

    Artemia has been used extensively in aquaculture as fodder for larval fish, shrimp, and shellfish. Epinecidin-1, an antimicrobial peptide, was isolated from grouper (Epinephelus coioides) in 2005. Epinecidin-1 has been previously reported to possess antimicrobial activity against several Gram-positive and Gram-negative bacterial species, including Staphylococcus coagulase, Pseudomonas aeruginosa, Streptococcus pyogenes, and Vibrio vulnificus. In this study, we used electroporation to introduce plasmid DNA encoding a green fluorescent protein (EGFP)-epinecidin-1 fusion protein under the control of the cytomegalovirus (CMV) promoter into decapsulated Artemia cysts. Optimization of various properties (including cyst weight (0.2 g), plasmid concentration (50 μg/100 μl), and pulse voltage (150 V), length (10 ms), and number (2)) resulted in a hatching rate of 41.15%, a transfection efficiency of 49.81%, and a fluorescence intensity (A.U.) of 47.46. The expression of EGFP-epinecidin-1 was first detected by quantitative RT-PCR at 120 h post-electroporation, and protein was identified by Western blot at the same time. Furthermore, the EGFP-epinecidin-1 protein inhibited V. vulnificus (204) growth, as demonstrated by zone of inhibition studies. Zebrafish fed on transgenic Artemia expressing CMV-gfp-epi combined with commercial fodder were more resistant to infection by V. vulnificus (204): survival rate was enhanced by over 70% at 7, 14, and 21 days post-infection, and bacterial numbers in the liver and intestine were reduced. In addition, feeding of transgenic Artemia to zebrafish affected the immunomodulatory response to V. vulnificus (204) infection; expression of immune-responsive genes, including hepcidin and defbl2, was altered, as shown by qPCR. These findings suggest that feeding transgenic Artemia expressing CMV-gfp-epi to larval fish has antimicrobial effects, without the drawbacks of introducing drug residues or inducing bacterial drug resistance

  2. Solution structure and dynamics of C-terminal regulatory domain of Vibrio vulnificus extracellular metalloprotease

    Energy Technology Data Exchange (ETDEWEB)

    Yun, Ji-Hye; Kim, Heeyoun [Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of); Park, Jung Eun [Department of Biotechnology, College of Natural Sciences, Chosun University, Gwangju 501-759 (Korea, Republic of); Lee, Jung Sup, E-mail: jsplee@mail.chosun.ac.kr [Department of Biotechnology, College of Natural Sciences, Chosun University, Gwangju 501-759 (Korea, Republic of); Lee, Weontae, E-mail: wlee@spin.yonsei.ac.kr [Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of)

    2013-01-11

    Highlights: Black-Right-Pointing-Pointer We have determined solution structures of vEP C-terminal regulatory domain. Black-Right-Pointing-Pointer vEP C-ter100 has a compact {beta}-barrel structure with eight anti-parallel {beta}-strands. Black-Right-Pointing-Pointer Solution structure of vEP C-ter100 shares its molecular topology with that of the collagen-binding domain of collagenase. Black-Right-Pointing-Pointer Residues in the {beta}3 region of vEP C-ter100 might be important in putative ligand/receptor binding. Black-Right-Pointing-Pointer vEP C-ter100 interacts strongly with iron ion. -- Abstract: An extracellular metalloprotease (vEP) secreted by Vibrio vulnificus ATCC29307 is a 45-kDa proteolytic enzyme that has prothrombin activation and fibrinolytic activities during bacterial infection. The action of vEP could result in clotting that could serve to protect the bacteria from the host defense machinery. Very recently, we showed that the C-terminal propeptide (C-ter100), which is unique to vEP, is involved in regulation of vEP activity. To understand the structural basis of this function of vEP C-ter100, we have determined the solution structure and backbone dynamics using multidimensional nuclear magnetic resonance spectroscopy. The solution structure shows that vEP C-ter100 is composed of eight anti-parallel {beta}-strands with a unique fold that has a compact {beta}-barrel formation which stabilized by hydrophobic and hydrogen bonding networks. Protein dynamics shows that the overall structure, including loops, is very rigid and stabilized. By structural database analysis, we found that vEP C-ter100 shares its topology with that of the collagen-binding domain of collagenase, despite low sequence homology between the two domains. Fluorescence assay reveals that vEP C-ter100 interacts strongly with iron (Fe{sup 3+}). These findings suggest that vEP protease might recruit substrate molecules, such as collagen, by binding at C-ter100 and that vEP participates

  3. The Fish Pathogen Vibrio vulnificus Biotype 2: Epidemiology, Phylogeny, and Virulence Factors Involved in Warm-Water Vibriosis.

    Science.gov (United States)

    Amaro, Carmen; Sanjuán, Eva; Fouz, Belén; Pajuelo, David; Lee, Chung-Te; Hor, Lien-I; Barrera, Rodolfo

    2015-06-01

    Vibrio vulnificus biotype 2 is the etiological agent of warm-water vibriosis, a disease that affects eels and other teleosts, especially in fish farms. Biotype 2 is polyphyletic and probably emerged from aquatic bacteria by acquisition of a transferable virulence plasmid that encodes resistance to innate immunity of eels and other teleosts. Interestingly, biotype 2 comprises a zoonotic clonal complex designated as serovar E that has extended worldwide. One of the most interesting virulence factors produced by serovar E is RtxA13, a multifunctional protein that acts as a lethal factor for fish, an invasion factor for mice, and a survival factor outside the host. Two practically identical copies of rtxA13 are present in all biotype 2 strains regardless of the serovar, one in the virulence plasmid and the other in chromosome II. The plasmid also contains other genes involved in survival and growth in eel blood: vep07, a gene for an outer membrane (OM) lipoprotein involved in resistance to eel serum and vep20, a gene for an OM receptor specific for eel-transferrin and, probably, other related fish transferrins. All the three genes are highly conserved within biotype 2, which suggests that they are under a strong selective pressure. Interestingly, the three genes are related with transferable plasmids, which emphasizes the role of horizontal gene transfer in the evolution of V. vulnificus in nutrient-enriched aquatic environments, such as fish farms.

  4. Phaeobacter inhibens as biocontrol agent against Vibrio vulnificus in oyster models

    DEFF Research Database (Denmark)

    Porsby, Cisse Hedegaard; Gram, Lone

    2016-01-01

    , the presence of P. inhibens could not prevent subsequently added V. vulnificus from entering the live animals, likely because of too low levels of the biocontrol strain. Whilst the oyster model studies provided indication that P. inhibens DSM 17395 could be a good candidate as biocontrol agent against V...

  5. Improved isolation of Vibro vulnificus from seawater and sediment with cellobiose-colistin agar

    DEFF Research Database (Denmark)

    Høi, L.; Dalsgaard, Inger; Dalsgaard, A.

    1998-01-01

    An improved selective medium, cellobiose-colistin (CC) agar, gave a significantly higher (P agar, In a total of 446 alkaline peptone water preenrichments amended...... with polymyxin B, V. vulnificus was isolated from 154 preenrichments (35%) with mCPC agar and from 179 preenrichments (40%) with CC agar. CC agar gave a higher plating efficiency of V. vulnificus cells than did cellobiose-polymyxin B-colistin (CPC) agar, mCPC agar, or thiosulfate-citrate-bile salts-sucrose (TCBS......) agar; the only significant difference was observed with TCBS agar, which gave much lower plating efficiencies than the other selective media. Determination of MICs demonstrated that the concentrations of colistin and polymyxin B in CPC agar inhibit growth of a proportion of V. vulnificus strains....

  6. Functional conservation of RNase III-like enzymes: studies on a Vibrio vulnificus ortholog of Escherichia coli RNase III.

    Science.gov (United States)

    Lee, Minho; Ahn, Sangmi; Lim, Boram; Lee, Dong-Ho; Lee, Kangseok

    2014-04-01

    Bacterial ribonuclease III (RNase III) belongs to the RNase III enzyme family, which plays a pivotal role in controlling mRNA stability and RNA processing in both prokaryotes and eukaryotes. In the Vibrio vulnificus genome, one open reading frame encodes a protein homologous to E. coli RNase III, designated Vv-RNase III, which has 77.9 % amino acid identity to E. coli RNase III. Here, we report that Vv-RNase III has the same cleavage specificity as E. coli RNase III in vivo and in vitro. Expressing Vv-RNase III in E. coli cells deleted for the RNase III gene (rnc) restored normal rRNA processing and, consequently, growth rates of these cells comparable to wild-type cells. In vitro cleavage assays further showed that Vv-RNase III has the same cleavage activity and specificity as E. coli RNase III on RNase III-targeted sequences of corA and mltD mRNA. Our findings suggest that RNase III-like proteins have conserved cleavage specificity across bacterial species.

  7. Faraday cage-type electrochemiluminescence immunosensor for ultrasensitive detection of Vibrio vulnificus based on multi-functionalized graphene oxide.

    Science.gov (United States)

    Guo, Zhiyong; Sha, Yuhong; Hu, Yufang; Yu, Zhongqing; Tao, Yingying; Wu, Yanjie; Zeng, Min; Wang, Sui; Li, Xing; Zhou, Jun; Su, Xiurong

    2016-10-01

    A novel Faraday cage-type electrochemiluminescence (ECL) immunosensor devoted to the detection of Vibrio vulnificus (VV) was fabricated. The sensing strategy was presented by a unique Faraday cage-type immunocomplex based on immunomagnetic beads (IMBs) and multi-functionalized graphene oxide (GO) labeled with (2,2'-bipyridine)(5-aminophenanthroline)ruthenium (Ru-NH2). The multi-functionalized GO could sit on the electrode surface directly due to the large surface area, abundant functional groups, and good electronic transport property. It ensures that more Ru-NH2 is entirely caged and become "effective," thus improving sensitivity significantly, which resembles extending the outer Helmholtz plane (OHP) of the electrode. Under optimal conditions, the developed immunosensor achieves a limit of detection as low as 1 CFU/mL. Additionally, the proposed immunosensor with high sensitivity and selectivity can be used for the detection of real samples. The novel Faraday cage-type method has shown potential application for the diagnosis of VV and opens up a new avenue in ECL immunoassay. Graphical abstract Faraday cage-type immunoassay mode for ultrasensitive detection by extending OHP.

  8. A novel recombinant bivalent outer membrane protein of Vibrio vulnificus and Aeromonas hydrophila as a vaccine antigen of American eel (Anguilla rostrata).

    Science.gov (United States)

    SongLin, Guo; PanPan, Lu; JianJun, Feng; JinPing, Zhao; Peng, Lin; LiHua, Duan

    2015-04-01

    The immogenicity of a novel vaccine antigen was evaluated after immunized American eels (Anguilla rostrata) with a recombinant bivalent expressed outer membrane protein (OMP) of Vibrio vulnificus and Aeromonas hydrophila. Three groups of eels were intraperitoneal (i.p) injected with phosphate-buffered saline (PBS group), formaline-killed-whole-cell (FKC) of A. hydrophila and V. vulnificus (FKC group) or the bivalent OMP (OMP group). On 14, 21, 28 and 42 days post-vaccination respectively, proliferation of the whole blood cells, titers of specific antibody and lysozyme activities of experimental eels were detected. On 28 day post-vaccination, eels from three groups were challenged by i.p injection of live A. hydrophila or V. vulnificus. The results showed that, compared with the PBS group, proliferation of whole blood cells in OMP group was significant enhanced on 28 days, and the serum titers of anti-A.hydrophila and anti-V. vulnificus antibody in eels of FKC and OMP group were significant increased on 14, 21 and 28d. Lysozyme Activities in serum, skin mucus, liver and kidney were significant changed between the three groups. Relative Percent Survival (RPS) after challenged A. hydrophila in KFC vs. PBS group and OMP vs. PBS group were 62.5% and 50% respectively, and the RPS challenged V. vulnificus in FKC and OMP vs. PBS group were 37.5% and 50% respectively. These results suggest that American eels immunized with the bivalent OMP would positively affect specific as well as non-specific immune parameters and protect against infection by the two pathogens in fresh water farming. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Antibiotic Susceptibility Patterns of Vibrio cholerae isolates

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    S D Shrestha

    2010-09-01

    Full Text Available INTRODUCTION: Cholera is one of the most common diarrhoeal diseases in Nepal. Etiological agent of cholera is Vibrio cholerae which removes essential body fluids, salts and vital nutrients, which are necessary for life causing dehydration and malnutrition. Emerging antimicrobial resistant is common. The aim of the present study was to determine the antibiotic susceptibility pattern of cholera patients in Nepal. METHODS: All the laboratory works were conducted in the bacteriology section of National Public Health Laboratory, Teku from March to September 2005. During this period a total of 340 stool samples from diarrhoeal patients were collected and processed according to the standard laboratory methods. Each patient suffering from diarrhoea was directly interviewed for his or her clinical history during sample collection. RESULTS: A total of 340 stool samples were processed and studied from both sex including all ages of patients. Among the processed sample 53 Vibrio cholerae cases were found. All isolated Vibrio cholerae O1 were El Tor, Inaba. All isolated (100% Vibrio cholerae O1 were sensitive to Ampicillin, Ciprofloxacin, Erythromycin and Tetracycline whereas all were resistant to Nalidixic acid and Cotrimoxazole. Only 15.1% cases were sensitive to Furazolidone whereas 84.9% were resistant. CONCLUSION: All V. cholerae strains isolated in this study were found resistant to Multi Drug Resistant (resistant to at least two antibiotics of different group. Ampicillin, Ciprofloxacin, Erythromycin and Tetracycline were found still more potent antibiotics against Vibrio cholerae isolated during the study. Keywords: antibiotics, susceptibility, Vibrio cholera.

  10. Use of a marker plasmid to examine differential rates of growth and death between clinical and environmental strains of Vibrio vulnificus in experimentally infected mice.

    Science.gov (United States)

    Starks, Angela M; Bourdage, Keri L; Thiaville, Patrick C; Gulig, Paul A

    2006-07-01

    Vibrio vulnificus is Gram-negative bacterium that contaminates oysters, causing highly lethal sepsis after consumption of raw oysters and wound infection. We previously described two sets of V. vulnificus strains with different levels of virulence in subcutaneously inoculated iron dextran-treated mice. Both virulent, clinical strains and attenuated, environmental strains could be recovered in high numbers from skin lesions and livers; however, the attenuated environmental strains required significantly higher numbers of colony-forming units (cfu) in the inoculum to produce lethal infection. Using some of these strains and an additional clinical strain, we presently asked if the different abilities to cause infection between the clinical and environmental strains were due to differences in rates of growth or death of the bacteria in the mouse host. We therefore constructed a marker plasmid, pGTR902, that functions as a replicon only in the presence of arabinose, which is not present in significant levels in animal tissues. V. vulnificus strains containing pGTR902 were inoculated into iron dextran-treated and untreated mice. Measuring the proportion of bacteria that had maintained the marker plasmid recovered from mice enabled us to monitor the number of in vivo divisions, hence growth rate; whereas measuring the number of marker plasmid-containing bacteria recovered enabled the measurement of death of the vibrios in the mice. The numbers of bacterial divisions in vivo for all of the strains over a 12-15 h infection period were not significantly different in iron dextran-treated mice; however, the rate of death of one environmental strain was significantly higher compared with the clinical strains. Infection of non-iron dextran-treated mice with clinical strains demonstrated that the greatest effect of iron dextran-treatment was increased growth rate, while one clinical strain also experienced increased death in untreated mice. V. vulnificus inoculated into iron

  11. Comparison of the Effects of Environmental Parameters on Growth Rates of Vibrio vulnificus Biotypes I, II, and III by Culture and Quantitative PCR Analysis ▿

    Science.gov (United States)

    Chase, Eva; Harwood, Valerie J.

    2011-01-01

    Vibrio vulnificus is a natural inhabitant of estuarine waters. The three known biotypes include (i) most human pathogens, (ii) primarily eel pathogens, and (iii) pathogens associated with fish and with human wound infections in Israel. Despite the frequently lethal consequences of V. vulnificus infections, the growth rates of the various biotypes and their response to environmental changes are not well characterized. We compared the specific growth rates (μ) of a representative of each biotype by culture and quantitative PCR (qPCR) analysis in a defined medium under varied pH, temperature, and salinity. Growth rates based on culturable concentrations were always higher than those based on qPCR estimates; however, both enumeration methods yielded comparable results on the influence of environmental factors on growth rates. Temperature (25°C, 30°C, 37°C), pH (7.0, 8.0), and salinity (5 to 40‰) all had significant effects on the μ of each biotype. Temperature had the greatest effect on the μ of biotype 1 (CMCP6), whereas salinity had the greatest effect on the μ of biotypes 2 (ATCC 33147) and 3 (302/99). The biotypes' growth rates varied significantly; biotype 1 grew most rapidly, while biotype 3 grew most slowly. The highest growth rates were achieved at 37°C, pH 7.0, and salinities of 15 to 30‰ (μ = 4.0, 2.9, and 2.4 generations h−1 for biotypes 1, 2, and 3, respectively). Other strains of the biotypes yielded comparable results, suggesting that the physiological responses of the biotypes are differentially affected by parameters that are highly variable both in estuarine environments and between the free-living and pathogen states of V. vulnificus. PMID:21515718

  12. Effects of Pollution on Vibrios in Woji River OJESANMI, A S; IBE, S N ...

    African Journals Online (AJOL)

    MICHAEL HORSFALL

    ABSTRACT: The effect of pollution on Vibrio spp. in five sampling stations along Woji River in Port. Harcourt was studied in the months of April and November 2010. Vibrio vulnificus, V. parahaemolyticus and V. alginolyticus were isolated. The Plate count technique on Thiosulphate Citrate Bile Salt agar revealed a high.

  13. Effects of Pollution on Vibrios in Woji River | Ojesanmi | Journal of ...

    African Journals Online (AJOL)

    The effect of pollution on Vibrio spp. in five sampling stations along Woji River in Port Harcourt was studied in the months of April and November 2010. Vibrio vulnificus, V. parahaemolyticus and V. alginolyticus were isolated. The Plate count technique on Thiosulphate Citrate Bile Salt agar revealed a high population density ...

  14. Effects of temperature and salinity on prevalence and intensity of infection of blue crabs, Callinectes sapidus, by Vibrio cholerae, V. parahaemolyticus, and V. vulnificus in Louisiana.

    Science.gov (United States)

    Sullivan, Timothy J; Neigel, Joseph E

    2018-01-01

    Coastal marine and estuarine environments are experiencing higher average temperatures, greater frequency of extreme temperature events, and altered salinities. These changes are expected to stress organisms and increase their susceptibility to infectious diseases. However, beyond these generalities, little is known about how environmental factors influence host-pathogen relationships in the marine realm. We investigated the prevalence and intensity of infections by Vibrio cholerae, V. parahaemolyticus, and V. vulnificus in blue crabs, Callinectes sapidus, from Louisiana saltmarshes in relation to temperature and salinity. We evaluated relationships for single measurements taken at the time of collection and for more complex measurements representing accumulated exposure to physiologically-stressful environmental conditions for up to 31 days prior to collection. We found that: (1) prevalence of infection varied across the Louisiana coast, (2) prevalence of all three Vibrio species was influenced by temperature and salinity, and (3) measurements that represent accumulated exposure to extreme conditions are useful predictors of infection prevalence and can provide insights into underlying biological mechanisms. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya.

    Science.gov (United States)

    Azwai, S M; Alfallani, E A; Abolghait, S K; Garbaj, A M; Naas, H T; Moawad, A A; Gammoudi, F T; Rayes, H M; Barbieri, I; Eldaghayes, I M

    2016-01-01

    The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk). Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6%) yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS) with culture characteristics of Vibrio spp. More than half (n=27) of processed seafood samples (n=46) yielded colonies on TCBS, while only 44.6 % of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 ×10(4) CFU\\g. Chicken burger samples showed the highest bacterial count with 6.5 ×10(4) CFU\\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9%) were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products.

  16. Isolation and molecular identification of Vibrio spp. by sequencing of 16S rDNA from seafood, meat and meat products in Libya

    Directory of Open Access Journals (Sweden)

    S.M. Azwai

    2016-03-01

    Full Text Available The genus Vibrio includes several food-borne pathogens that cause a spectrum of clinical conditions including septicemia, cholera and milder forms of gastroenteritis. Several Vibrio spp. are commonly associated with food-borne transmission including Vibrio cholerae, Vibrio parahemolyticus, and Vibrio vulnificus. Microbiological analysis for enumeration and isolation of Vibrio spp. were carried out for a total of 93 samples of seafood, meat and meat products from different geographic localities in Libya (Tripoli, Regdalin, Janzour and Tobruk. Vibrio spp. were detected by conventional cultural and molecular method using PCR and sequencing of 16S rDNA. Out of the 93 cultured samples only 48 (51.6% yielded colonies on Thiosulfate Citrate Bile Salt agar (TCBS with culture characteristics of Vibrio spp. More than half (n=27 of processed seafood samples (n=46 yielded colonies on TCBS, while only 44.6% of samples of meat and meat products showed colonies on TCBS. Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 х104 CFU\\g. Chicken burger samples showed the highest bacterial count with 6.5 х104 CFU\\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9% were Vibrio spp. Vibrio parahemolyticus was isolated from camel meat for the first time. This study is an initial step to provide a baseline for future molecular research targeting Vibrio spp. foodborne illnesses. This data will be used to provide information on the magnitude of such pathogens in Libyan seafood, meat and meat products.

  17. Genome sequencing and analysis of a highly virulent Vibrio parahaemolyticus strain isolated from the marine environment

    Science.gov (United States)

    Parks, M. C.; Moreno, E.

    2016-02-01

    Vibrio parahaemolyticus [Vp] is a Gram-negative bacterium and a natural inhabitant of coastal marine ecosystems worldwide. Vp is also a coincidental pathogen of humans. Virulent strains are commonly identified by the presence of the thermostable direct (tdh) or tdh-related (trh) hemolysin genes. However, virulence is multifaceted and many clinical Vp isolates do not carry tdh or trh. In this study, we sequenced and assembled the draft genome of a tdh- and trh-negative environmental isolate (805) shown previously to be highly virulent in zebrafish. To investigate potential mechanisms of virulence, we compared 805 to the clinical V. parahaemolyticus type strain (RIMD2210633). Pairwise comparison revealed the presence of multiple genomic regions including an IncF conjugative pilus (1.3 Kb) and a colicin V plasmid (1.49 Kb). These features are homologous to genomic regions present in clinical V. vulnificus and V. cholerae strains. Genome comparison also revealed the presence of five toxin-antitoxin systems. Isolate 805 likely attained these new features through the lateral acquisition of mobile genomic material - a hypothesis supported by the aberrant GC content of these regions. Colicin V plasmids are a diverse group of IncF plasmids found in invasive bacterial strains. Similarly, an abundance of toxin-antitoxin systems have been linked to virulence in Gram-negative bacteria. Current efforts are focused on characterizing 142 coding features present in 805 but absent from the type strain.

  18. Antibiotic susceptibility profiles of some Vibrio strains isolated from wastewater final effluents in a rural community of the Eastern Cape Province of South Africa

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    Igbinosa Etinosa O

    2010-05-01

    Full Text Available Abstract Background To evaluate the antibiogram and antibiotic resistance genes of some Vibrio strains isolated from wastewater final effluents in a rural community of South Africa. V. vulnificus (18, V. metschnikovii (3, V. fluvialis (19 and V. parahaemolyticus (12 strains were isolated from final effluents of a wastewater treatment plant (WWTP located in a rural community of South Africa. The disk diffusion method was used for the characterization of the antibiogram of the isolates. Polymerase chain reaction (PCR was employed to evaluate the presence of established antibiotic resistance genes using specific primer sets. Results The Vibrio strains showed the typical multidrug-resistance phenotype of an SXT element. They were resistant to sulfamethoxazole (Sul, trimethoprim (Tmp, cotrimoxazole (Cot, chloramphenicol (Chl, streptomycin (Str, ampicillin (Amp, tetracycline (Tet nalidixic acid (Nal, and gentamicin (Gen. The antibiotic resistance genes detected includes dfr18 and dfrA1 for trimethoprim; floR, tetA, strB, sul2 for chloramphenicol, tetracycline, streptomycin and sulfamethoxazole respectively. Some of these genes were only recently described from clinical isolates, demonstrating genetic exchange between clinical and environmental Vibrio species. Conclusions These results demonstrate that final effluents from wastewater treatment plants are potential reservoirs of various antibiotics resistance genes. Moreover, detection of resistance genes in Vibrio strains obtained from the wastewater final effluents suggests that these resistance determinants might be further disseminated in habitats downstream of the sewage plant, thus constituting a serious health risk to the communities reliant on the receiving waterbodies.

  19. Isolation and molecular identification of Vibrio spp. by sequencing of ...

    African Journals Online (AJOL)

    Among cultured seafood samples, the highest bacterial count was recorded in clam with a count of 3.8 х104 CFU\\g. Chicken burger samples showed the highest bacterial count with 6.5 х104 CFU\\g. Molecular analysis of the isolates obtained in this study, showed that 11 samples out of 48 (22.9%) were Vibrio spp. Vibrio ...

  20. Model for end-stage liver disease (MELD score as a predictor and monitor of mortality in patients with Vibrio vulnificus necrotizing skin and soft tissue infections.

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    Kuo-Chin Huang

    2015-04-01

    Full Text Available Vibrio vulnificus necrotizing skin and soft tissue infections (VNSSTIs usually predispose patients with or without preexisting liver disease to septic shock, and then evolve to multiple organ dysfunction syndrome (MODS, thus resulting in high mortality in humans. However, clinicians do not have a valid prediction model to provide a reliable estimate of case-fatality rate when caring for these acutely and/or critically ill patients.We retrospectively analyzed 39 consecutive patients with VNSSTIs (mean age: 65.7 ± 11.3 years at our institution between 2007 and 2010. All patients were treated with the same protocol. Demographic and clinical characteristics, disease severity on admission, treatment details, and outcomes were collected for each patient and extracted for analyses. We studied the predictive value of the model for end-stage liver disease (MELD, modified MELD including sodium (MELD-Na, and laboratory risk indicator for necrotizing fasciitis (LRINEC scores for case-fatality. Logistic regression and receiver operating characteristic (ROC curve analyses were performed. The mean MELD, MELD-Na and LRINEC scores on admission were 15.1 ± 1.1, 17.7 ± 1.1, and 3.4 ± 0.4 points, respectively. After admission, these patients had temporary or progressive deterioration of nearly all their scores and lab values. The area under the ROC curve for the MELD and ΔMELD scoring models were 0.929 (p = 0.002 and 0.897 (p = 0.005, respectively. An optimal MELD/ΔMELD cutoff value ≥ 20/2 had a good sensitivity and specificity (all > 80%, with a 64/13-fold increased odds for case-fatality. Additionally, the development of severe forms of anemia (p = 0.014 and hypoalbuminemia (p = 0.019 were associated with an increased case-fatality rate.The MELD/ΔMELD scoring model is an effective risk stratification indicator at the time of admission and also an excellent condition monitor during hospitalization for medical care of acutely and/or critically ill patients

  1. MR imaging findings of lower extremity sepsis caused by vibro vulnificus: A report of three cases

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    Kim, Jihyun; Koh, Sung Hye; Min, Soo Kee; Lee, Jeong A; Lee, Kwan Seop [Hallym University College of Medicine, Hallym University Sacred Heart Hospital, Anyang (Korea, Republic of)

    2017-01-15

    Lower extremity infection caused by Vibrio vulnificus sepsis is a rapidly progressing fatal condition. Prompt diagnosis followed by early and aggressive treatment with antibiotics and fasciotomy is crucial. In this report, we described lower extremity magnetic resonance (MR) images of three patients with Vibrio vulnificus sepsis. In our cases, MR imaging of lower extremity with Vibrio vulnificus sepsis showed three common findings. First, the MR signal abnormalities appeared simultaneously in all layers, including skin, subcutaneous fat, muscles, and deep fasciae. Second, the inflammation showed symmetry on both legs. Third, none of our cases was accompanied by abscess formation. These imaging features may represent rapid progression of Vibrio vulnificus sepsis and could be helpful for accurate diagnosis, and prompt and aggressive treatment.

  2. Antibiotic Resistance of Vibrio cholerae Isolates from Kashan, Iran

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    Afzali H.MD,

    2016-03-01

    Full Text Available Abstract Aims: Cholera is an acute diarrheal disease that can lead to severe dehydration and death. Antibiotic resistance is a big challenge in infective disease like Cholera. The present study aimed to understand the characteristics and trends of antibiotic resistance of V. cholerae isolations in and around Kashan, Iran. Instrument & Methods: In this descriptive cross-sectional study, samples were gathered using census method from 1998 to 2013 in Kashan, Iran. 1132 fecal samples of patients with acute diarrhea and 237 samples of suspected water samples were taken. The serotypes and biotypes were determined by an enzymatic method. Antibiotic susceptibility test was performed by using Disk Diffusion Method. Data were analyzed using SPSS 23 software. Fisher-exact and Chi-square tests were used to compare the statistical parameters. Findings: 96 fecal samples (8.5% and 18 water samples (7.6% were positive for Vibrio cholerae. Non-agglutinating (Nag isolates (75.4% were more common than serotype Inaba (13.2% and Ogawa (11.4%. Nag serotypes were mostly resistant to cefixime (44% and ampicillin (33%. In contaminated water samples also the most frequent cases were Nag serotype (50%. Nag serotype showed 22.2% of resistance to ampicillin and nitrofurantoin. Conclusion: Vibrio cholerae isolates in Kashan, Iran, are highly resistant to antibiotics, especially Nag serotypes.

  3. Vibrio spp. isolados a partir de mexilhões (Perna perna in natura e pré-cozidos de Estação Experimental de Cultivo, Rio de Janeiro, RJ, Brasil Vibrio spp. isolated from in natura and precooked mussels (Perna perna of an Experimental Station Culture, Rio de Janeiro, RJ, Brazil

    Directory of Open Access Journals (Sweden)

    Christiane Soares Pereira

    2007-06-01

    Full Text Available A análise microbiológica dos mexilhões reflete a qualidade do habitat aquático, pois estes animais podem reter em seus organismos diversos patógenos, dentre os quais aqueles pertencentes à família Vibrionaceae. No presente estudo foi avaliada a presença de Vibrio spp. em mexilhões (in natura e pré-cozidos, comercializados na Estação Experimental de Cultivo de Mexilhões, situada em Jurujuba, Niterói, Rio de Janeiro. Foram avaliadas 86 amostras, tomando como procedimento, o enriquecimento em Água Peptonada Alcalina (APA adicionada de 1 e 3% de NaCl, isolamento em Agar Tiossulfato Citrato Bile Sacarose (TCBS e confirmação das colônias típicas por análise bioquímica. Dentre as 12 espécies de Vibrio identificadas destacaram-se como de maior prevalência as espécies Vibrio alginolyticus, V. cholerae não-O1, V. parahaemolyticus, V. carchariae e Vibrio vulnificus. A relevância epidemiológica destes patógenos associada a casos de gastrenterite humana após consumo de mexilhões crus ou parcialmente cozidos, reforça a importância de alertar as autoridades de Vigilância Sanitária sobre sua presença na cadeia alimentar e seus riscos para a Saúde Pública.The microbiology analysis of mussels reflects the aquatic environment quality because these animals retain various pathogens such as Vibrionaceae family microorganisms in their organism. In the present investigation, we evaluated the presence of Vibrio spp. in mussels (in natura and precooked commercialized at an Experimental Station Mussel Culture in Jurujuba, Niterói, Rio de Janeiro. A total of 86 samples were analyzed using the enrichment in Alkaline Peptone Water (APW added to 1 and 3% of NaCl, isolated in Thiossulfate Citrate Bile Sucrose Agar (TCBS and confirmed that there were suspect colonies by biochemical tests. The results showed the identification of 12 different Vibrio species whereby Vibrio alginolyticus, V. cholerae non-O1, V. parahaemolyticus, V. carchariae

  4. Genetic characterization of trh positive Vibrio spp. isolated from Norway

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    Anette eBauer Ellingsen

    2013-12-01

    Full Text Available The thermostable direct hemolysin (TDH and/or TDH-related hemolysin (TRH genes are carried by most virulent Vibrio parahaemolyticus serovars. In Norway, trh+ V. parahaemolyticus constitute 4.4% and 4.5 % of the total number of V. parahaemolyticus isolated from blue mussel (Mytilus edulis and water, respectively. The trh gene is located in a region close to the gene cluster for urease production (ure. This region was characterized in V. parahaemolyticus strain TH3996 and it was found that a nickel transport operon (nik was located between the first gene (ureR and the rest of the ure cluster genes. The organization of the trh-ureR-nik-ure gene cluster in the Norwegian trh+ isolates was unknown. In this study, we explore the gene organization within the trh-ureR-nik-ure cluster for these isolates. PCR analyses revealed that the genes within the trh-ureR-nik-ure gene cluster of Norwegian trh+ isolates were organized in a similar fashion as reported previously for TH33996. Additionally, the phylogenetic relationship among these trh+ isolates was investigated using Multilocus Sequence Typing (MLST. Analysis by MLST or ureR-trh sequences generated two different phylogenetic trees for the same strains analyzed, suggesting that ureR-trh genes have been acquired at different times in Norwegian V. parahaemolyticus isolates. MLST results revealed that some pathogenic and non-pathogenic V. parahaemolyticus isolates in Norway appear to be highly genetically related.

  5. Genetic characterization of trh positive Vibrio spp. isolated from Norway.

    Science.gov (United States)

    Ellingsen, Anette B; Olsen, Jaran S; Granum, Per E; Rørvik, Liv M; González-Escalona, Narjol

    2013-01-01

    The thermostable direct hemolysin (TDH) and/or TDH-related hemolysin (TRH) genes are carried by most virulent Vibrio parahaemolyticus serovars. In Norway, trh+ V. parahaemolyticus constitute 4.4 and 4.5% of the total number of V. parahaemolyticus isolated from blue mussel (Mytilus edulis) and water, respectively. The trh gene is located in a region close to the gene cluster for urease production (ure). This region was characterized in V. parahaemolyticus strain TH3996 and it was found that a nickel transport operon (nik) was located between the first gene (ureR) and the rest of the ure cluster genes. The organization of the trh-ureR-nik-ure gene cluster in the Norwegian trh+ isolates was unknown. In this study, we explore the gene organization within the trh-ureR-nik-ure cluster for these isolates. PCR analyses revealed that the genes within the trh-ureR-nik-ure gene cluster of Norwegian trh+ isolates were organized in a similar fashion as reported previously for TH33996. Additionally, the phylogenetic relationship among these trh+ isolates was investigated using Multilocus Sequence Typing (MLST). Analysis by MLST or ureR-trh sequences generated two different phylogenetic trees for the same strains analyzed, suggesting that ureR-trh genes have been acquired at different times in Norwegian V. parahaemolyticus isolates. MLST results revealed that some pathogenic and non-pathogenic V. parahaemolyticus isolates in Norway appear to be highly genetically related.

  6. Vibrio fujianensis sp. nov., isolated from aquaculture water.

    Science.gov (United States)

    Fang, Yujie; Chen, Aiping; Dai, Hang; Huang, Ying; Kan, Biao; Wang, Duochun

    2018-02-13

    A Gram-stain-negative, facultatively anaerobic strain, designated FJ201301 T , was isolated from aquaculture water collected from Fujian province, China. Phylogenetic analysis of 16S rRNA gene sequences indicated that strain FJ201301 T belonged to the genus Vibrio, formed a distinct cluster with Vibriocincinnatiensis ATCC 35912 T and shared the highest similarity with Vibriosalilacus CGMCC 1.12427 T . A 15 bp insertion found in the 16S rRNA gene was a significant marker that distinguished strain FJ201301 T from several phylogenetic neighbours (e.g. V. cincinnatiensis). Multilocus sequence analysis of eight genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA; concatenated 4135 bp sequence) showed that, forming a long and independent phylogenetic branch, strain FJ201301 T clustered with V. cincinnatiensis ATCC 35912 T , Vibrioinjenensis KCTC 32233 T and Vibriometschnikovii CIP 69.14 T clearly separated from V. salilacus CGMCC 1.12427 T . Furthermore, the highest in silico DNA-DNA hybridization and average nucleotide identity values between strain FJ201301 T and the closest related species were 26.3 and 83.1 % with V. cincinnatiensis ATCC 35912 T , less than the proposed cutoff levels for species delineation, i.e. 70 and 95 %, respectively. Biochemical, sequence and genomic analysis suggested the designation of strain FJ201301 T representing a novel species of the genus Vibrio, for which the name Vibrio fujianensis sp. nov. is proposed. The type strain is FJ201301 T (=DSM 104687 T =CGMCC 1.16099 T ).

  7. Occurrences of pathogenic Vibrio parahaemolyticus from Vellar ...

    African Journals Online (AJOL)

    user

    2013-04-03

    2001). Vibrio vulnificus as a health hazard for shrimp consumers. Revista do Instituto de Medicina Tropical de Sao Paulo. 43: 263- 266. Sambrook J, Fritsch EF, Maniatis T (1989). Molecular Cloning: A. Laboratory Manual, 2nd ed.

  8. Influence of temperature, salinity and pH on the growth of environmental Aeromonas and Vibrio species isolated from Mai Po and the Inner Deep Bay Nature Reserve Ramsar Site of Hong Kong.

    Science.gov (United States)

    Wang, Yanling; Gu, Ji-Dong

    2005-01-01

    Four environmental bacterial isolates including Aeromonas hydrophila MP-3, A. salmonicida MP-4, Vibrio vulnificus MP-2 and V. cholerae MP-1 isolated from sediment and water of Mai Po Nature Reserve of Hong Kong were examined for their responses to temperature, pH and salinity under laboratory conditions in this study. V. cholerae MP-1 was found to resist vibriostatic agent O/129 at concentration of 10 microg/ml. In addition, bacterial growth under test conditions was measured and the results were fitted into the Gompertz model to obtain important parameters related to bacterial growth, lag time (lambda), specific growth rate (mu(m)), and maximum biomass (A) for comparison. V. cholerae MP-1 did not show any apparent growth at 15 degrees C, but was adapted to a much wider environmental pH from 5.2 to 9.2 for growth while V. vulnificus MP-2 was more sensitive to pH changes yielding the highest biomass at pH 6.2. A. salmonicida MP-4 was surprisingly tolerant to salinity as high as 60.0 per thousand NaCl and grew almost equally well as under conditions of other treatments. All four bacterial isolates showed a wide spectrum of plasticity to the environmental conditions and they pose a potential threat to public health and animal health. ((c) 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

  9. Prevalence and Antimicrobial Resistance of Vibrio spp. in Retail and Farm Shrimps in Ecuador.

    Science.gov (United States)

    Sperling, L; Alter, T; Huehn, S

    2015-11-01

    The aim of this study was to investigate the prevalence of Vibrio spp. in shrimp at retail and in shrimp farms in Ecuador and to determine the antimicrobial agent resistance patterns of farm isolates. The presence of genes linked to early mortality syndrome (EMS) or acute hepatopancreatic necrosis disease (AHPND) also was evaluated. Vibrio spp. were isolated from retail shrimps in Cuenca, Ecuador, and farm shrimps originating from provinces El Oro and Guayas, Ecuador. A total of 229 shrimp samples were collected, of which 71 originated from retail markets in Cuenca and 158 came from shrimp farms. Overall, 219 (95.6%) samples tested positive for Vibrio spp. Vibrio parahaemolyticus (80.8%) was the most common species detected, followed by Vibrio alginolyticus (50.2%), Vibrio cholerae (11.3%), and Vibrio vulnificus (3.5%). None of the V. parahaemolyticus isolates carried the virulence-associated tdh and trh genes. In V. parahaemolyticus shrimp farm isolates, high resistance was found to ampicillin (92.2%), and intermediate resistance was found to tetracycline (51.3%) and amikacin (22.1%). Of the V. parahaemolyticus strains, 68 were resistant to at least three antimicrobial agents, and 2 were resistant to seven antimicrobial agents simultaneously. Up to 18 resistant isolates were found for V. alginolyticus, whereas V. vulnificus and V. cholerae isolates were more susceptible. None of the V. parahaemolyticus isolates carried the EMS-AHPND plasmid. The results of this study revealed the ubiquitous occurrence of Vibrio spp. in shrimps at retail and on shrimp farms in Ecuador.

  10. Genome sequence of vibrio cholerae G4222, a South African clinical isolate

    CSIR Research Space (South Africa)

    Le Rouw, Wouter J

    2013-03-01

    Full Text Available Vibrio cholerae, a Gram-negative pathogen autochthonous to the aquatic environment, is the causative agent of cholera. Here, we report the complete genome sequence of V. cholerae G4222, a clinical isolate from South Africa....

  11. Occurrence of Vibrio Pathotypes in the Final Effluents of Five Wastewater Treatment Plants in Amathole and Chris Hani District Municipalities in South Africa

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    Vuyokazi Nongogo

    2014-08-01

    Full Text Available We assessed the occurrence of Vibrio pathogens in the final effluents of five wastewater treatment plants (WWTPs located in Amathole and Chris Hani District Municipalities in South Africa over a 12 months period between September 2012 and August 2013 using standard membrane filtration technique followed by cultivation on thiosulphate citrate-bile salts-sucrose (TCBS agar. The identities of the presumptive Vibrio isolates were confirmed using polymerase chain reaction (PCR including delineation into V. parahaemolyticus, V. vulnificus and V. fluvialis pathotypes. The counts of Vibrio spp. varied with months in all the study sites and ranged in the order of 101 and 104 CFU/100mL. Vibrio distribution also showed seasonality with high counts being obtained in autumn and spring (p < 0.05. Prevalence of Vibrio spp. among the five WWTPs also differed significantly (p < 0.05. Of the 300 isolates that were confirmed as belonging to the Vibrio genus, 29% (86 were V. fluvialis, 28% (84 were V. vulnificus and 12% (35 were V. parahaemolyticus. The isolation of Vibrio pathogens from the final effluent suggests that this pathogen is in circulation in some pockets of the population and that the WWTPs under study do not efficiently remove bacterial pathogens from the wastewater and consequently are threats to public health.

  12. Occurrence and Diversity of Clinically Important Vibrio Species in the Aquatic Environment of Georgia.

    Science.gov (United States)

    Kokashvili, Tamar; Whitehouse, Chris A; Tskhvediani, Ana; Grim, Christopher J; Elbakidze, Tinatin; Mitaishvili, Nino; Janelidze, Nino; Jaiani, Ekaterine; Haley, Bradd J; Lashkhi, Nino; Huq, Anwar; Colwell, Rita R; Tediashvili, Marina

    2015-01-01

    Among the more than 70 different Vibrio species inhabiting marine, estuarine, and freshwater ecosystems, 12 are recognized as human pathogens. The warm subtropical climate of the Black Sea coastal area and inland regions of Georgia likely provides a favorable environment for various Vibrio species. From 2006 to 2009, the abundance, ecology, and diversity of clinically important Vibrio species were studied in different locations in Georgia and across seasons. Over a 33-month period, 1,595 presumptive Vibrio isolates were collected from the Black Sea (n = 657) and freshwater lakes around Tbilisi (n = 938). Screening of a subset of 440 concentrated and enriched water samples by PCR-electrospray ionization/mass spectrometry (PCR-ESI/MS) detected the presence of DNA from eight clinically important Vibrio species: V. cholerae, V. parahaemolyticus, V. vulnificus, V. mimicus, V. alginolyticus, V. harveyi, V. metschnikovii, and V. cincinnatiensis. Almost 90% of PCR/ESI-MS samples positive for Vibrio species were collected from June through November. Three important human-pathogenic Vibrio species (V. cholerae, V. parahaemolyticus, and V. vulnificus) were detected in 62.8, 37.8, and 21.4% of samples testing positive for Vibrios, respectively. The results of these activities suggest that natural reservoirs for human-pathogenic Vibrios exist in Georgian aquatic environments. Water temperature at all sampling sites was positively correlated with the abundance of clinically important Vibrio spp. (except V. metschnikovii), and salinity was correlated with species composition at particular Black Sea sites as well as inland reservoirs.

  13. Antagonistic effect of Pseudomonas aeruginosa isolates from various ecological niches on Vibrio species pathogenic to crustaceans

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    Prabhakaran Priyaja

    2014-01-01

    Full Text Available Objective: To abrogate pathogenic vibrios in aquaculture by testing the potential of Pseudomonas isolates from fresh water, brackish and marine environments as probiotics. Methods: Purification and structural elucidation of antagonistic compound were carried out. Antagonistic activity of the compound against 7 Vibrio spp. was performed. Influence of salinity on the production of pyocyanin and the toxicity was done through the compound using brine shrimp lethality assay. Molecular characterization was performed to confirm that the isolates were Pseudomonas aeruginosa. Results: Salinity was found to regulate the levels of pyocyanin production, with 5-10 g/L as the optimum. All Pseudomonas isolates grew at salinities ranging from 5 to 70 g/L. Isolates of marine origin produced detectable levels of pyocyanin up to 45 g/L salinity. Brackish and freshwater isolates ceased to produce pyocyanin at salinities above 30 g/L and 20 g/L, respectively. Culture supernatants of all 5 Pseudomonas isolates possessed the ability to restrict the growth of Vibrio spp. and maximum antagonistic effect on Vibrio harveyi was obtained when they were grown at salinities of 5 to 10 g/L. The marine isolate MCCB117, even when grown at a salinity of 45 g/L possessed the ability to inhibit Vibrio spp. Conclusions: The present investigation showed that Pseudomonas aeruginosa MCCB119 would be ideal for application in freshwater, MCCB102 and MCCB103 in brackish water and MCCB117 and MCCB118 in marine aquaculture systems as putative probiotics in the management of vibrios.

  14. Monitoring of different vibrio species affecting marine fishes in Lake Qarun and Gulf of Suez: Phenotypic and molecular characterization

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    Mohamed Abdelaziz

    2017-06-01

    Full Text Available Vibriosis is a globally threatening bacterial disease affecting mariculture with high mortalities and severe economic losses. Isolation and Identification of different vibrio species were performed to a total number of one hundred moribund and freshly dead Solea aegyptiaca, Epinephelus marginatus and Mugil cephalus collected from Lake Qarun and Gulf of Suez. Phenotypic picture and molecular identification based on use of 16SrRNA gene sequence confirmed 44 strains as vibrio species. Further molecular identification of retrieved vibrio spp. using species specific primers for collagenase, ToxR and Vvh genes categorized 10 isolates belong to V. alginolyticus, 8 isolates belong to V. parahaemolyticus and 6 isolates belong to V. vulnificus. The total prevalence of vibriosis was 44% where the highest prevalence was recorded in Lake Qarun examined fishes.

  15. Temporal and spatial variability in culturable pathogenic Vibrio spp. in Lake Pontchartrain, Louisiana, following hurricanes Katrina and Rita.

    Science.gov (United States)

    Nigro, Olivia D; Hou, Aixin; Vithanage, Gayatri; Fujioka, Roger S; Steward, Grieg F

    2011-08-01

    We investigated the abundance, distribution, and virulence gene content of Vibrio cholerae, V. parahaemolyticus, and V. vulnificus in the waters of southern Lake Pontchartrain in Louisiana on four occasions from October 2005 to September 2006, using selective cultivation and molecular assays. The three targeted pathogenic vibrios were generally below the detection level in January 2006, when the water was cold (13°C), and most abundant in September 2006, when the lake water was warmest (30°C). The maximum values for these species were higher than reported previously for the lake by severalfold to orders of magnitude. The only variable consistently correlated with total vibrio abundance within a single sampling was distance from shore (P = 0.000). Multiple linear regression of the entire data set revealed that distance from shore, temperature, and turbidity together explained 82.1% of the variability in total vibrio CFU. The log-transformed mean abundance of V. vulnificus CFU in the lake was significantly correlated with temperature (P = 0.014), but not salinity (P = 0.625). Virulence-associated genes of V. cholerae (ctx) and V. parahaemolyticus (trh and tdh) were not detected in any isolates of these species (n = 128 and n = 20, respectively). In contrast, 16S rRNA typing of V. vulnificus (n = 298) revealed the presence of both environmental (type A) and clinical (type B) strains. The percentage of the B-type V. vulnificus was significantly higher in the lake in October 2005 (35.8% of the total) than at other sampling times (P ≤ 0.004), consistent with the view that these strains represent distinct ecotypes.

  16. Isolation of Vibrio parahaemolyticus from fecal specimens on mannitol salt agar.

    Science.gov (United States)

    Carruthers, M M; Kabat, W J

    1976-08-01

    Unless laboratories use an inhibitory medium, Vibrio parahaemolyticus will be unrecognizable in fecal specimens. The use of a medium exclusively for vibrio isolation, such as thiosulfate-citrate-bile salts-sucrose agar (TCBS), however, may not be considered economically justified in the United States. The isolation and recognition of V. parahaemolyticus is reported on mannitol salt agar (MS), a medium which is used for fecal specimens here. Eight Kanagawa-positive and two of three Kanagawa-negative strains of V. parahaemolyticus grew as well on MS as on TCBS and better than on a representative enteric medium, Hektoen enteric agar (HE). Twenty-two fecal specimens from 16 noninfected individuals were inoculated with known quantities of V. parahaemolyticus, and recovery of these vibrios was assessed on TCBS, MS, and HE. Recovery of vibrios from MS and TCBS was similar when inoculum size was 10(3) colony-forming units/ml or greater. Recovery of vibrios from mixed culture was distinctly lower on HE. The colonial morphology of V. parahaemolyticus and several other bacteria on MS is illustrated.

  17. Differential specificity of selective culture media for enumeration of pathogenic vibrios: advantages and limitations of multi-plating methods.

    Science.gov (United States)

    Nigro, Olivia D; Steward, Grieg F

    2015-04-01

    Plating environmental samples on vibrio-selective chromogenic media is a commonly used technique that allows one to quickly estimate concentrations of putative vibrio pathogens or to isolate them for further study. Although this approach is convenient, its usefulness depends directly on how well the procedure selects against false positives. We tested whether a chromogenic medium, CHROMagar Vibrio (CaV), used alone (single-plating) or in combination (double-plating) with a traditional medium thiosulfate-citrate-bile-salts (TCBS), could improve the discrimination among three pathogenic vibrio species (Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus) and thereby decrease the number of false-positive colonies that must be screened by molecular methods. Assays were conducted on water samples from two estuarine environments (one subtropical, one tropical) in a variety of seasonal conditions. The results of the double-plating method were confirmed by PCR and 16S rRNA sequencing. Our data indicate that there is no significant difference in the false-positive rate between CaV and TCBS when using a single-plating technique, but determining color changes on the two media sequentially (double-plating) reduced the rate of false positive identification in most cases. The improvement achieved was about two-fold on average, but varied greatly (from 0- to 5-fold) and depended on the sampling time and location. The double-plating method was most effective for V. vulnificus in warm months, when overall V. vulnificus abundance is high (false positive rates as low as 2%, n=178). Similar results were obtained for V. cholerae (minimum false positive rate of 16%, n=146). In contrast, the false positive rate for V. parahaemolyticus was always high (minimum of 59%, n=109). Sequence analysis of false-positive isolates indicated that the majority of confounding isolates are from the Vibrionaceae family, however, members of distantly related bacterial groups were also able to

  18. Vibrio galatheae sp. nov., a novel member of the Vibrionaceae family isolated from the Solomon Sea

    DEFF Research Database (Denmark)

    Giubergia, Sonia; Machado, Henrique; Mateiu, Ramona Valentina

    2015-01-01

    Based on genetic, chemotaxonomic and phenotypic characteristics, a novel species belonging to the genus Vibrio is described. The facultative anaerobic strain S2757T was isolated from a mussel collected in the Solomon Sea (Solomon Islands). Phylogenetic analyses based on sequences of 16S rRNA and ...

  19. Prevalence and heterogeneity of Hemolysin gene vhh among hatchery isolates of Vibrio harveyi in India

    Digital Repository Service at National Institute of Oceanography (India)

    Parvathi, A.; George, J.; Kumar, S.

    , Baticados MCL, Cruz-Lacierda ER, de la Pena EL (1990) Occurrence of luminous bacterial disease of Penaeus monodon larvae in the Philippines. Aquaculture 91:1-13 19. Liu PC, Lee KK, Chen SN (1996) Pathogenicity of different isolates of Vibrio harveyi...

  20. Characterization of a Vibrio cholerae phage isolated from the coastal water of Peru.

    Science.gov (United States)

    Talledo, Miguel; Rivera, Irma N G; Lipp, Erin K; Neale, Angela; Karaolis, David; Huq, Anwar; Colwell, Rita R

    2003-05-01

    A Vibrio cholerae bacteriophage, family Myoviridae, was isolated from seawater collected from the coastal water of Lima, Peru. Genome size was estimated to be 29 kbp. The temperate phage was specific to V. cholerae and infected 12/13 V. cholerae O1 strains and half of the four non-O1/non-O139 strains tested in this study. Vibrio cholerae O139 strains were resistant to infection and highest infection rates were obtained in low nutrient media amended with NaCl or prepared using seawater as diluent.

  1. Vibrio japonicus sp. nov., a novel member of the Nereis clade in the genus Vibrio isolated from the coast of Japan.

    Science.gov (United States)

    Doi, Hiroyasu; Osawa, Ikuko; Adachi, Hayamitsu; Kawada, Manabu

    2017-01-01

    A novel Vibrio strain, JCM 31412T, was isolated from seawater collected from the Inland Sea (Setonaikai), Japan, and characterized as a Gram-negative, oxidase-positive, catalase-negative, facultatively anaerobic, motile, ovoid-shaped bacterium with one polar flagellum. Based on 16S rDNA gene identity, strain JCM 31412T showed a close relationship with type strains of Vibrio brasiliensis (LMG 20546T, 98.2% identity), V. harveyi (NBRC 15634T, 98.2%), V. caribbeanicus (ATCC BAA-2122T, 97.8%) and V. proteolyticus (NBRC 13287T, 97.8%). The G+C content of strain JCM 31412T DNA was 46.8%. Multi-locus sequence analysis (MLSA) of eight loci (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA; 5535bp) further clustered strain JCM 31412T in the Nereis clade, genus Vibrio. Phenotypically, strain JCM 31412T differed from the closest related Vibrio species in its utilization of melibiose and raffinose, and its lack of casein and gelatin hydrolysis. It was further differentiated based on its fatty acid composition, specifically properties of C12:03OH and summed features, which were significantly different from those of V. brasiliensis, V. nigripulchritudo and V. caribbeanicus type strains. Overall, the results of DNA-DNA hybridization, and physiological and biochemical analysis differentiated strain JCM 31412T from other described species of the genus Vibrio. Based on these polyphasic taxonomic findings, it was therefore concluded that JCM 31412T was a novel Vibrio species, for which the name Vibrio japonicus sp. nov. was proposed, with JCM 31412T (= LMG 29636T = ATCC TSD-62T) as the type strain.

  2. Vibrio japonicus sp. nov., a novel member of the Nereis clade in the genus Vibrio isolated from the coast of Japan.

    Directory of Open Access Journals (Sweden)

    Hiroyasu Doi

    Full Text Available A novel Vibrio strain, JCM 31412T, was isolated from seawater collected from the Inland Sea (Setonaikai, Japan, and characterized as a Gram-negative, oxidase-positive, catalase-negative, facultatively anaerobic, motile, ovoid-shaped bacterium with one polar flagellum. Based on 16S rDNA gene identity, strain JCM 31412T showed a close relationship with type strains of Vibrio brasiliensis (LMG 20546T, 98.2% identity, V. harveyi (NBRC 15634T, 98.2%, V. caribbeanicus (ATCC BAA-2122T, 97.8% and V. proteolyticus (NBRC 13287T, 97.8%. The G+C content of strain JCM 31412T DNA was 46.8%. Multi-locus sequence analysis (MLSA of eight loci (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA and topA; 5535bp further clustered strain JCM 31412T in the Nereis clade, genus Vibrio. Phenotypically, strain JCM 31412T differed from the closest related Vibrio species in its utilization of melibiose and raffinose, and its lack of casein and gelatin hydrolysis. It was further differentiated based on its fatty acid composition, specifically properties of C12:03OH and summed features, which were significantly different from those of V. brasiliensis, V. nigripulchritudo and V. caribbeanicus type strains. Overall, the results of DNA-DNA hybridization, and physiological and biochemical analysis differentiated strain JCM 31412T from other described species of the genus Vibrio. Based on these polyphasic taxonomic findings, it was therefore concluded that JCM 31412T was a novel Vibrio species, for which the name Vibrio japonicus sp. nov. was proposed, with JCM 31412T (= LMG 29636T = ATCC TSD-62T as the type strain.

  3. PREVALENCE AND IDENTIFICATION OF VIBRIO SPP. ISOLATED ON AQUACULTURED GILTHEAD SEA BREAM

    Directory of Open Access Journals (Sweden)

    C. Scarano

    2011-01-01

    Full Text Available The aim of the study was to investigate the prevalence of Vibrio spp isolated from gilthead sea bream (Sparus aurata farmed on sea cages and to identify and characterize the pathogen by molecular techniques. Eighty fish were collected from two hatcheries located on the North-Est Sardinian Mediterranean coast, and microbiological analysis were performed on different body parts such as skin, gills, muscle and intestinal tract. Subsequently 100 pure colonies with typical morphology and phenotypic characteristics were selected and submitted to the molecular identification. The analysis on the prevalence of Vibrio spp showed the effect of the hatchery rearing system (P<0.001, of the date of sampling (P<0.001, and of the body part (P<0.001. All the strains selected were confirmed to be members of the genus Vibrio spp by the molecular method/techinique/identification, whereas the rpoA gene sequence analyses allowed to identify 89 strains belonging to the species Vibrio harveyi, 6 to V. diabolicus, 2 to V. parahaemolyticus and 1 to V. mediterranei.

  4. [Pathogenicity island region of clinical and environmental strains of Vibrio parahaemolyticus, isolated in Chile].

    Science.gov (United States)

    Núñez, Harold; Ulloa, María Teresa; Guerra, Fabiola; Osorio, Carlos G

    2009-02-01

    Most clinical isolates of Vibrio parahaemolyticus produce a major virulence factor known as the thermostable direct hemolysin (TDH). TDH is encoded by the tdh gene which is located in a genomic pathogenicity island (PAI). Most environmental isolates are described as tdh negative. To assess if environmental strains lack the full pathogenicity island or if only the tdh gene is deleted. Thirty eight clinical and 66 environmental strains of Vibrio parahaemolyticus were studied. PAI was characterized by polymerase chain reaction (PCR). The presence of tdhA and tdhS genes, was determined by Southern blot. Fifty three environmental strains (80%) lacked a full PAI when compared with clinical strains. In environmental strains, Southern blot and sequence analysis showed that a genetic region of 80 kilobase pairs including genes from VPA1310 to VPA1396 was missing. These results highlight the genetic dynamism of Vibrio parahaemolyticus pathogenecity island region and suggest that new pathogenic strains could appear by horizontal transfer of the island between toxigenic and non-toxigenic strains.

  5. [ISOLATION OF ANTIBIOTICS RESISTANCE GENES IN VIBRIO CHOLERAE O1 AND O139 SEROGROUP STRAINS].

    Science.gov (United States)

    Zadnova, S P; Smirnova, N I

    2015-01-01

    Determination of sensitivity of V. cholerae O1 serogroup El Tor biovar and O139 serogroup strains to antibiotics and determination of the presence of antibiotics resistance genes in their genome. The studies were carried out in 75 V. cholerae O1 and O139 serogroup strains. Sensitivity of cultures to antibiotics was determined by disc-diffusion method. DNA isolation was carried out in the presence of 6M guanidine thiocyanate. PCR was carried out in multi-channel amplificator Tercyc. A multiplex PCR was constructed, that includes 5 primer pairs for the detection of O1 and O139 serogroup resistance genes of vibrios to sulfame- thoxazolum, streptomycin B, trimethoprim, the presence of SXT element, an amplification program was developed. Using the developed PCR, V. cholerae O1 serogroup El Tor biovar strains with multiple drug resistance were established to be imported into Russia in 1993. The presence of SXT elements with genes of resistance to 4 antibiotics simultaneously was detected precisely in these strains, that belong to toxigenic genovariants of V. cholerae El Tor biovar. All the El Tor vibrio strains imported in the subsequent years were shown to stably preserve SXT element, this indicates its important role in biology of cholera vibrios. O139 serogroup strains with intact SXT element and having a deletion of the gene coding trimethoprim resistance were isolated. The data obtained may be used to establish molecular-genetic mechanisms of emergence of antibiotics resistant strains of cholera vibrio, construction of novel gene diagnostic test-systems and carrying out passportization of strains that are stored in the State collection of pathogenic bacteria.

  6. Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from retail shrimps in Malaysia

    Directory of Open Access Journals (Sweden)

    Vengadesh eLetchumanan

    2015-01-01

    Full Text Available Vibrio parahaemolyticus is a marine and estuarine bacterium that has been the leading cause of foodborne outbreaks which leads to a significant threat to human health worldwide. Consumption of seafood contaminated with Vibrio parahaemolyticus causes acute gastroenteritis in individuals. The bacterium poses two main virulence factor including the thermostable direct hemolysin (tdh which is a pore-forming protein that contributes to the invasiveness of the bacterium in humans and TDH-related hemolysin (trh, which plays a similar role as thermostable direct hemolysin (tdh in the disease pathogenesis. This study aimed to investigate the antimicrobial resistance Vibrio parahaemolyticus strains in shrimps purchased from wetmarkets and supermarkets. The toxR-based PCR assay indicated that a total of 57.8% (185/320 isolates were positive for V. parahaemolyticus. Only 10% (19/185 toxR-positive isolate exhibit the TDH-related hemolysin (trh gene and none of the isolates were tested positive for thermostable direct hemolysin (tdh. The MAR index was measured for 14 common antimicrobial agents. The results indicated 98% of the isolates were highly susceptible to imipenem, ampicillin sulbactam (96%, chloramphenicol (95%, trimethoprim-sulfamet (93%, gentamicin (85%, levofloxacin (83% and tetracycline (82%. The chloramphenicol (catA2 and kanamycin (aphA-3 resistance genes were detected in the resistant V. parahaemolyticus isolates. Our results demonstrate that shrimps are contaminated with V. parahaemolyticus, some of which carry the trh-gene thus being potential to cause food borne illness. The occurrence of multidrug resistance strains in the environment could be an indication of excessive usage of antibiotics in agriculture and aquaculture fields.

  7. Pathogenetic characterization of Vibrio parahaemolyticus isolates from clinical and seafood sources.

    Science.gov (United States)

    Vongxay, Khamphouth; Wang, Shuna; Zhang, Xiaofeng; Wu, Beibei; Hu, Hongxia; Pan, Zijiang; Chen, Suyun; Fang, Weihuan

    2008-08-15

    A total of 216 Vibrio parahaemolyticus isolates from seafood and clinical samples in eastern China were investigated for their hemolytic and urea-producing phenotypes, presence of putative virulence genes tdh and trh. Twenty-one clinical isolates (84%, 21/25) and 3 seafood isolates (1.57%, 3/191) were tdh-positive while only 3 clinical isolates (12%) and 7 seafood isolates (3.66%) were positive for trh gene. We further examined the pathogenicity of selected V. parahaemolyticus isolates in in vitro and in vivo systems. The clinical isolates were apparently more enteropathogenic (74.26 per thousand vs 62.07 per thousand expressed as intestine/body weight ratio, Pcytotoxicity as measured by LDH release of the HeLa cells although there were no statistical differences. The tdh-positive V. parahaemolyticus isolates were of higher enteropathogenicity (Pcytotoxic and adhesive to the cultured cell lines as well. From the in vitro and in vivo pathogenicity profiles, trh-positive isolates seemed to line between tdh-positive isolates and those without tdh and trh. There were two isolates H8 and H10 from clinical cases having moderate enteropathogenicity and virulence to mice, but were tdh-negative yet trh-positive. These results seem to suggest that hemolysins TDH and/or TRH may not be necessarily the only virulence factors of pathogenic V. parahaemolyticus isolates.

  8. Population dynamics of Vibrio and Pseudomonas species isolated from farmed Tasmanian Atlantic salmon (Salmo salar L.): a seasonal study.

    Science.gov (United States)

    Hatje, Eva; Neuman, Christina; Stevenson, Hollie; Bowman, John P; Katouli, Mohammad

    2014-11-01

    Vibrio and Pseudomonas species have been shown to be part of the normal microbiota of Atlantic salmon (Salmo salar L.), with some strains causing disease in fish. The factors affecting their prevalence and persistence in the salmon gut, however, have not been well studied. In this study, we collected 340 Vibrio and 150 Pseudomonas isolates from the hindgut of farmed Tasmanian Atlantic salmon, fed with two commercially available diets. Samples were collected every 6-8 weeks between July 2011 and May 2012. Isolates from selective agar were initially identified using biochemical tests and confirmed using genus-specific primers and 16S ribosomal RNA (16S rRNA) sequencing. Random amplified polymorphic DNA (RAPD) PCR was used to type both Pseudomonas and Vibrio; the latter was further typed using a biochemical fingerprinting method (PhP-RV plates). We observed low species diversity with strains comprising Vibrio ichthyoenteri/Vibrio scophthalmi, Vibrio crassostreae/Vibrio splendidus, Aliivibrio finisterrensis, Photobacterium phosphoreum and Pseudomonas fragi. Out of 340 Vibrio isolates, 238 (70 %) belonged to 21 clonal types and were found predominantly during summer when water temperatures reached 15 to 21 °C. Of these, the four major clonal types were found in multiple samples (70 %). P. fragi, on the other hand, was only found during the colder water temperatures and belonged to 18 clonal types. The presence of both groups of bacteria and their clonal types were independent of the fish diets used, suggesting that the water temperature was the main factor of the prevalence and persistence of these bacteria in the gut of Atlantic salmon.

  9. Vibrio zhanjiangensis sp. nov., isolated from sea water of shrimp farming pond.

    Science.gov (United States)

    Jin, Chunying; Luo, Peng; Zuo, Huali; Chen, Jianming; Chen, Mingliang; Wang, Wei

    2012-05-01

    A Gram-negative, facultatively anaerobic, motile by means of single polar flagellum, rod-shaped marine bacterium, designated strain E414, was isolated from sea water collected from a farming pond rearing marine shrimp Litopenaeus vannamei in Zhanjiang, Guangdong province, PRC. The strain was able to grow in the presence of 0.5-6% (w/v) NaCl (optimally in 3-6% (w/v) NaCl), between pH 6 and 9 (optimally at pH 7-8), between 15 and 37°C (optimally at 25-30°C). Phylogenetic analysis based on 16S rRNA gene sequences locate strain E414 in the vicinity of the coralliilyticus clade within the genus Vibrio. DNA-DNA relatedness data and multigene phylogenetic analysis based on the concatenated sequences of four genes (16S rRNA, rpoA, recA and pyrH) clearly differentiated strain E414 from its closest phylogenetic neighbours. Analysis of phenotypic features, including enzyme activities and utilization and fermentation of various carbon sources, further revealed discrimination between strain E414 and phylogenetically related Vibrio species. The major fatty acid components are C(16:1)ω6c and/or C(16:1)ω7c (27.4%), C(18:1)ω7c and/or C(18:1)ω6c (19.3%) and C(16:0) (18.2%). The DNA G+C content of strain E414 was 38.7 mol%. Based on phenotypic, chemotaxonomic, phylogenetic and DNA-DNA relatedness values, it can be concluded that E414 should be placed in the genus Vibrio as representing a novel species, for which the name Vibrio zhanjiangensis sp. nov. is proposed, with the type strain E414 (=CCTCC AB 2011110(T) = NBRC 108723(T) = DSM 24901).

  10. Detection, isolation, and identification of Vibrio cholerae from the environment.

    Science.gov (United States)

    Huq, Anwar; Haley, Bradd J; Taviani, Elisa; Chen, Arlene; Hasan, Nur A; Colwell, Rita R

    2012-08-01

    Recent molecular advances in microbiology have greatly improved the detection of bacterial pathogens in the environment. These improvements and a downward trend in the cost of molecular detection methods have contributed to increased frequency of detection of pathogenic microorganisms where traditional culture-based detection methods have failed. Culture methods also have been greatly improved, and the confluence of the two suites of methods provides a powerful tool for detection, isolation, and characterization of pathogens. While molecular detection provides data on the presence and type of pathogens, culturing methods allow a researcher to preserve the organism of interest for "-omics" studies, such as genomic, metabolomic, secretomic, and transcriptomic analysis, which are rapidly becoming more affordable. This has yielded a clearer understanding of the ecology and epidemiology of microorganisms that cause disease. In this unit, we present commonly accepted methods for isolation, detection, and characterization of V. cholerae, providing more extensive knowledge of the ecology and epidemiology of this organism. This unit has been fully revised and updated from the earlier version with the latest knowledge and additional information not previously included. © 2012 by John Wiley & Sons, Inc.

  11. Characterization of a new beta-lactamase gene from isolates of Vibrio spp. in Korea.

    Science.gov (United States)

    Jun, Lyu Jin; Kim, Jae Hoon; Jin, Ji Woong; Jeong, Hyun Do

    2012-04-01

    PCR was performed to analyze the beta-lactamase genes carried by ampicillin-resistant Vibrio spp. strains isolated from marine environments in Korea between 2006 and 2009. All 36 strains tested showed negative results in PCR with the primers designed from the nucleotide sequences of various known beta-lactamase genes. This prompted us to screen new beta-lactamase genes. A novel beta-lactamase gene was cloned from Vibrio alginolyticus KV3 isolated from the aquaculture water of Geoje Island of Korea. The determined nucleotide sequence (VAK-3 beta-lactamase) revealed an open reading frame (ORF) of 852 bp, encoding a protein of 283 amino acids (aa), which displayed low homology to any other beta-lactamase genes reported in public databases. The deduced 283 aa sequence of VAK-3, consisting of a 19 aa signal peptide and a 264 aa mature protein, contained highly conserved peptide segments specific to class A beta-lactamases including the specific amino acid residues STFK (62-65), SDN (122-124), E (158), and RTG (226-228). Results from PCR performed with primers specific to the VAK-3 beta-lactamase gene identified 3 of the 36 isolated strains as V. alginolyticus, Vibrio cholerae, and Photobacterium damselae subsp. damselae, indicating the utilization of various beta-lactamase genes including unidentified ones in ampicillin-resistant Vibrio spp. strains from the marine environment. In a mating experiment, none of the isolates transfered the VAK-3 beta-lactamase gene to the Escherichia coli recipient. This lack of mobility, and the presence of a chromosomal acyl-CoA flanking sequence upstream of the VAK-3 beta- lactamase gene, led to the assumption that the location of this new beta-lactamase gene was in the chromosome, rather than the mobile plasmid. Antibiotic susceptibility of VAK-3 beta-lactamase was indicated by elevated levels of resistance to penicillins, but not to cephalosporins in the wild type and E. coli harboring recombinant plasmid pKV-3, compared with those of

  12. Population dynamics, antibiotics resistance and biofilm formation of Aeromonas and Vibrio species isolated from aquatic sources in Northern Malaysia.

    Science.gov (United States)

    Odeyemi, Olumide A; Ahmad, Asmat

    2017-02-01

    This study aimed to compare population dynamics, antibiotic resistance and biofilm formation of Aeromonas and Vibrio species from seawater and sediment collected from Northern Malaysia. Isolates with different colony morphology were characterized using both biochemical and molecular methods before testing for antibiotic resistance and biofilm formation. Results obtained from this study showed that in Kedah, the population of Aeromonas isolated from sediment was highest in Pantai Merdeka (8.22 log CFU/ml), Pulau Bunting recorded the highest population of Aeromonas from sediment (8.43 log CFU/g). It was observed that Vibrio species isolated from seawater and sediment were highest in Kuala Sanglang (9.21 log CFU/ml). In Kuala Perlis, the population of Aeromonas isolated from seawater was highest in Jeti (7.94 log CFU/ml). Highest population of Aeromonas from sediment was recorded in Kampong Tanah Baru (7.99 log CFU/g). It was observed that Vibrio species isolated from seawater was highest in Padang Benta (8.42 log CFU/g) while Jeti Kuala Perlis had highest population of Vibrio isolated from sediment. It was observed that location does not influence population of Aeromonas. The results of the independent t - test revealed that there was no significant relationship between location and population of Vibrio (df = 10, t = 1.144, p > 0.05). The occurrence of biofilm formation and prevalence of antibiotic resistant Aeromonas and Vibrio species in seawater and sediment pose danger to human and aquatic animals' health. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from seafoods in Lagos Lagoon Nigeria

    Directory of Open Access Journals (Sweden)

    Chigozie Oramadike

    2015-12-01

    Full Text Available In this study, a total of 90 seafood samples; croaker fish (Pseudotolithus senegalensis, shrimps (Penaeus notialis and blue crab (Callinectes sapidus collected from landing sites along the Lagos Lagoon in Nigeria were examined for the prevalence of Vibrio parahaemolyticus using both biochemical and molecular methods. Biochemical identification of the isolates was confirmed by Polymerase Chain Reaction (PCR. The presence of the virulence-associated tdh (thermostable direct haemolysin, trh1 (thermostable-related haemolysin and trh2 genes in the V. parahaemolyticus isolates was also detected by the PCR method. PCR products from the V.16S primers were sequenced. Antibiotics susceptibility of the isolates was also determined. About, eight isolates were presumptively identified as V. parahaemolyticus, PCR identified five and none of the isolates were positive for the genes tdh or trh. The five isolates sequenced were identified as different strains of V. parahaemolyticus. V. parahaemolyticus_RIMD_2210633 = 2MKSHa remained resistant to all antimicrobials tested. However, only V. parahaemolyticus_MP-2_AY911391 = TBSHy showed strong sensitivity to all the antimicrobials with ampicillin (minimum inhibitory concentration-4 μg/ml. In addition, the other three isolates showed sensitivity for Tetracycline, Ciprofloxacin, Gentamicin and Ceftazidime. Ampicillin resistance in most of the isolates suggests low efficiency of ampicillin in management of V. parahaemolyticus infection.

  14. Isolation and characterisation of Bacillus spp. antagonistic to Vibrio parahaemolyticus for use as probiotics in aquaculture.

    Science.gov (United States)

    Liu, Xue-Fei; Li, Ya; Li, Jian-Rong; Cai, Lu-Yun; Li, Xiu-Xia; Chen, Jin-Ru; Lyu, Shu-Xia

    2015-05-01

    Acute gastroenteritis caused by pathogenic Vibrio parahaemolyticus is one of the major factors affecting the development of aquaculture and the safety of seafood. Using the antagonism of probiotics against pathogens is an alternative strategy to antibiotics and a common trend to control food-borne pathogenic bacteria. In this study, a total of 249 isolates were isolated from four types of seafood (Litopenaeus vannamei, Oratosquilla oratoria, Mactra veneriformis and Portunus trituberculatus) and coastal sediment from Liaodong Bay in the Bohai Sea, China with five different separation agars. The most isolates came from the sample of coastal sediment and on agar of 2216E, which accounted for 36.14 and 54.62 % respectively. Twenty-four among 249 isolates displayed direct antimicrobial activity to V. parahaemolyticus with spot inoculation. Sixteen active isolates were selected for extracellular antimicrobial activity using the Oxford cup method. Only strains of B16 and J7 showed extracellular antimicrobial activity and were identified as Bacillus pumilus and Bacillus mojavensis respectively based on the physiological identification and 16S rRNA sequence analysis. Both of the strains B16 and J7 exhibited extracellular hydrolytic enzyme activity and antagonism against more than one indicator bacteria in vitro, which indicates that the two strains have broad potential application as suitable probiotic candidates in aquaculture while B. mojavensis was first reported to inhibit pathogenic Vibrio spp. in vitro. There is no particular trait as to antagonism of B. pumilus B16 or B. mojavensis J7 to Gram-positive or Gram-negative indicator bacteria.

  15. Phenotypic and phylogenetic analysis of Vibrio parahaemolyticus isolates recovered from diarrhea cases in Guangdong Province, China

    Science.gov (United States)

    Li, Baisheng; Luo, Jinyan; Tan, Hailing; Ke, Bixia; He, Dongmei; Ke, Changwen; Klena, John D.; Zhang, Yonghui

    2015-01-01

    Vibrio parahaemolyticus has emerged as a common foodborne pathogen of global concern. In this study, 108 V. parahaemolyticus isolates that recovered from diarrhea cases (n = 96) and seafood products (n = 12) in Guangdong Province from 2007 to 2011 were characterized by serotyping, tdh and trh toxin gene detection and multilocus sequence typing (MLST). The dominant serotypes from the cases were O3:K6, O4:K8 and O1: KUT (untyped). However, most isolates recovered from seafood products belonged to other serotypes. None of the isolates carried the trh gene, while the major isolates from the cases were tdh positive. MLST analysis revealed 31 sequence types (STs); 17 STs were unique in this study. eBURST analysis revealed four clonal complexes (CC), The majority of the isolates (n = 58, all from cases and tdh+) were grouped into the CC3, which included O3:K6, O4:K68 and O1:KUT isolates. The CC3 was the most prevalent clonal complex, and all of the CC3 isolates were recovered from clinical cases of geographically diverse origin. As to the CC345, which was completely constituted by O4:K8, was another important clonal complex affecting Guangdong Province. Ongoing surveillance of V. parahaemolyticus in diarrhea patients and seafood products remains a public health priority for Guangdong Province, China. PMID:25662708

  16. Characterization of Vibrio Parahaemolyticus isolated from oysters and mussels in São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Martha Virginia Ribeiro Rojas

    2011-08-01

    Full Text Available Vibrio parahaemolyticus is a marine bacterium, responsible for gastroenteritis in humans. Most of the clinical isolates produce thermostable direct hemolysin (TDH and TDH-related hemolysin (TRH encoded by tdh and trh genes respectively. In this study, twenty-three V. parahaemolyticus, previously isolated from oysters and mussels were analyzed by PCR using specific primers for the 16S rRNA and virulence genes (tdh, trh and tlh and for resistance to different classes of antibiotics and PFGE. Nineteen isolates were confirmed by PCR as V. parahaemolyticus. The tlh gene was present in 100% of isolates, the tdh gene was identified in two (10.5% isolates, whereas the gene trh was not detected. Each isolate was resistant to at least one of the nine antimicrobials tested. Additionally, all isolates possessed the blaTEM-116 gene. The presence of this gene in V. parahaemolyticus indicates the possibility of spreading this gene in the environment. Atypical strains of V. parahaemolyticus were also detected in this study.

  17. Differences in the API 20E biochemical patterns of clinical and environmental Vibrio parahaemolyticus isolates.

    Science.gov (United States)

    Martinez-Urtaza, Jaime; Lozano-Leon, Antonio; Viña-Feas, Alejandro; de Novoa, Jacobo; Garcia-Martin, Oscar

    2006-02-01

    Genetic differences in clinical and environmental strains of Vibrio parahaemolyticus have been widely used as criteria in identifying pathogenic isolates. However, few studies have been carried out to assess the differences in biochemical characteristics of V. parahaemolyticus isolates from human and environmental sources. We compared the biochemical profiles obtained by the characterization of V. parahaemolyticus isolates from human infections and the marine environment using the API 20E system. Environmental and clinical isolates showed significant differences in the gelatin and arabinose tests. Additionally, clinical isolates were correctly identified according to the API 20E profile using 0.85% NaCl diluent, but they presented nonspecific profiles with 2% NaCl diluent. In contrast, use of 2% NaCl diluent facilitated correct identification of the environmental isolates. Clinical isolates showed significant differences in up to five biochemical tests with respect to the API 20E database. The API 20E system is widely used in routine identification of bacteria in clinical laboratories, and this discrepancy in an important number of biochemical tests may lead to misidentification of V. parahaemolyticus infection.

  18. Comparative Genomics of Vibrio cholerae O1 Isolated from Cholera Patients in Bangladesh

    DEFF Research Database (Denmark)

    Hossain, Zenat Zebin; Leekitcharoenphon, Pimlapas; Dalsgaard, Anders

    AIM: Cholera remains an endemic disease in Bangladesh and recently, the severity of the disease has significantly increased in urban area since the emergence of the new variant of Vibrio cholerae O1 El Tor. In this study, Whole Genome Sequencing (WGS) was utilized to investigate the current genomic...... profile of V. cholerae O1 strains, isolated from symptomatic patients in the low-income urban area of Arichpur, Dhaka, Bangladesh. METHODS: During October 2015, three V. cholerae O1 strains (VC-1, 2 and 3) were isolated from rectal swabs of two patients living in households 588 m apart. One of the two...... patients was co-infected with two V. cholerae strains (VC-1 and VC-3). Major virulence factors, biotype and antimicrobial resistance genes were identified by WGS. A global phylogenetic tree was inferred using genome wide SNPs (Single Nucleotide Polymorphism) analysis. RESULTS: All the V. cholerae strains...

  19. Urea hydrolysis can predict the potential pathogenicity of Vibrio parahaemolyticus strains isolated in the Pacific Northwest.

    Science.gov (United States)

    Kaysner, C A; Abeyta, C; Trost, P A; Wetherington, J H; Jinneman, K C; Hill, W E; Wekell, M M

    1994-01-01

    The ability of some strains of Vibrio parahaemolyticus to hydrolyze urea (uh+) can be used as a marker to predict which strains isolated from molluscan shellfish harvested in the Pacific Northwest are potentially pathogenic. The thermostable direct hemolysin-producing (TDH+) characteristic is a marker that is correlated with potential pathogenicity, and all of the TDH+ strains that we have isolated have been found to be uh+. Most of the uh+ strains belong to somatic antigen groups O3, O4 and O5. TDH+ strains are usually members of groups O4 and O5. The strains most often associated with human illness are members of the uh+, O4 group. The test for urease production is a simple screening test that can be helpful in predicting which strains are potentially pathogenic. PMID:8085837

  20. Genomic Features of Environmental and Clinical Vibrio parahaemolyticus Isolates Lacking Recognized Virulence Factors Are Dissimilar

    Science.gov (United States)

    Petronella, N.; Chew Leung, C.; Pightling, A. W.; Banerjee, S. K.

    2015-01-01

    Vibrio parahaemolyticus is a bacterial pathogen that can cause illness after the consumption or handling of contaminated seafood. The primary virulence factors associated with V. parahaemolyticus illness are thermostable direct hemolysin (TDH) and Tdh-related hemolysin (TRH). However, clinical strains lacking tdh and trh have recently been isolated, and these clinical isolates are poorly understood. To help understand the emergence of clinical tdh- and trh-negative isolates, a genomic approach was used to comprehensively compare 4 clinical tdh- and trh-negative isolates with 16 environmental tdh- and trh-negative isolates and 34 clinical isolates positive for tdh or trh, or both, with the objective of identifying genomic features that are unique to clinical tdh- and trh-negative isolates. The prevalence of pathogenicity islands (PAIs) common to clinical isolates was thoroughly examined in each of the clinical tdh- and trh-negative isolates. The tdh PAI was not present in any clinical or environmental tdh- and trh-negative isolates. The trh PAI was not present in any environmental isolates; however, in clinical tdh- and trh-negative isolate 10-4238, the majority of the trh PAI including a partial trh1 gene was present, which resulted in reclassification of this isolate as a tdh-negative and trh-positive isolate. In the other clinical tdh- and trh-negative isolates, neither the trh gene nor the trh PAI was present. We identified 862 genes in clinical tdh- and trh-negative isolates but not in environmental tdh- and trh-negative isolates. Many of these genes are highly homologous to genes found in common enteric bacteria and included genes encoding a number of chemotaxis proteins and a novel putative type VI secretion system (T6SS) effector and immunity protein (T6SS1). The availability of genome sequences from clinical V. parahaemolyticus tdh- and trh-negative isolates and the comparative analysis may help provide an understanding of how this pathotype is able to

  1. Onderzoek pathogene vibrio soorten in Nederlandse mosselen en oesters in augustus en september 2003

    NARCIS (Netherlands)

    Aalberts, C.H.J.

    2003-01-01

    Naar aanleiding van de hoge weerstemperatuur in augustus 2003 zijn in de kweek- en verwatergebieden van mosselen en oesters in Nederland enkele monsters onderzocht op de aanwezigheid van voor de mens pathogene vibrio soorten. In geen van de 18 monsters is Vibrio parahaemolyticus, vulnificus of

  2. Vibrio tapetis isolated from vesicular skin lesions in Dover sole Solea solea.

    Science.gov (United States)

    Declercq, A M; Chiers, K; Soetaert, M; Lasa, A; Romalde, J L; Polet, H; Haesebrouck, F; Decostere, A

    2015-06-29

    Vibrio tapetis is primarily known as the causative agent for brown ring disease in bivalves, although it has been isolated from cultivated fish during mortalities on farms. Here we describe the first isolation of V. tapetis from wild-caught and subsequently captive-held Dover sole Solea solea. Pathological features consisted of multifocal circular greyish-white skin discolourations evolving into vesicular lesions and subsequent ulcerations on the pigmented side. On the non-pigmented side, multiple circular lesions-white at the center and red at the edges-were evident. Histological examination of the vesicular lesions revealed dermal fluid-filled spaces, collagen tissue necrosis and a mixed inflammatory infiltrate, with large numbers of small rod-shaped bacteria. In the deep skin lesions, loss of scales and dermal connective tissue, with degeneration and fragmentation of the myofibres bordering the ulceration, were noted. Serotyping, DNA-DNA hybridization and REP- and ERIC-PCR techniques showed that the retrieved isolates displayed a profile similar to the representative strain of genotype/serotype O2 which originally was isolated from carpet-shell clam Venerupis decussata and to which isolates obtained from wedge sole Dicologoglossa cuneata were also closely related.

  3. AP4 method for two-tube nested PCR detection of AHPND isolates of Vibrio parahaemolyticus

    Directory of Open Access Journals (Sweden)

    Sirintip Dangtip

    2015-11-01

    Full Text Available Our previous work on the mechanism of virulence for the unique isolates of Vibrio parahaemolyticus that cause acute hepatopancreatic necrosis disease (VPAHPND revealed that it was mediated by a binary Pir-like toxin pair ToxA and ToxB. These toxins are located on the pVA plasmid, a plasmid carried by AHPND-causing strain of V. parahaemolyticus with a size of approximately 69 kbp. Using the coding sequences of ToxA, a one-step PCR detection method for VPAHPND was introduced in June 2014 but had the limitation that attempts to adapt it into a nested PCR protocol were unsuccessful. As a result, low levels of VPAHPND in shrimp or other samples could not be detected without first preparing an enrichment broth culture to allow bacterial growth before extraction of template DNA. Here, we describe the AP4 (abbreviation of AHPND detection version 4 method, a two-tube nested PCR method that targets the tandem genes ToxA and ToxB, including the 12 bp spacer that separates them on pVA plasmid. Testing of the method revealed that it gave 100% positive and negative predictive values for VPAHPND using a panel of 104 bacterial isolates including 51 VPAHPND isolates and 53 non-AHPND isolates, the latter including 34 isolates of V. parahaemolyticus and 19 isolates of other bacteria found in shrimp ponds, including other Vibrio species. The AP4 nested PCR method was 100 times more sensitive (100 fg total DNA template than the one-step AP3 (10 pg total DNA template method, and it could detect VPAHPND in experimentally challenged shrimp by 6 h post immersion (n = 2/3, while AP3 could not detect is until 12 h post immersion (n = 1/3. Thus, the AP4 method may be useful in detecting VPAHPND isolates in samples where target material is limited (e.g., small tissue quantity or archived DNA and enrichment cannot be employed (i.e., frozen samples or samples preserved in alcohol.

  4. Vibrio aphrogenes sp. nov., in the Rumoiensis clade isolated from a seaweed.

    Science.gov (United States)

    Tanaka, Mami; Endo, Shoko; Kotake, Fumihito; Al-Saari, Nurhidayu; Amin, A K M Rohul; Feng, Gao; Mino, Sayaka; Doi, Hidetaka; Ogura, Yoshitoshi; Hayashi, Tetsuya; Suda, Wataru; Hattori, Masahira; Yumoto, Isao; Sawabe, Toko; Sawabe, Tomoo; Araki, Toshiyoshi

    2017-01-01

    A novel strain Vibrio aphrogenes sp. nov. strain CA-1004T isolated from the surface of seaweed collected on the coast of Mie Prefecture in 1994 [1] was characterized using polyphasic taxonomy including multilocus sequence analysis (MLSA) and a genome based comparison. Both phylogenetic analyses on the basis of 16S rRNA gene sequences and MLSA based on eight protein-coding genes (gapA, gyrB, ftsZ, mreB, pyrH, recA, rpoA, and topA) showed the strain could be placed in the Rumoiensis clade in the genus Vibrio. Sequence similarities of the 16S rRNA gene and the multilocus genes against the Rumoiensis clade members, V. rumoiensis, V. algivorus, V. casei, and V. litoralis, were low enough to propose V. aphrogenes sp. nov. strain CA-1004T as a separate species. The experimental DNA-DNA hybridization data also revealed that the strain CA-1004T was separate from four known Rumoiensis clade species. The G+C content of the V. aphrogenes strain was determined as 42.1% based on the genome sequence. Major traits of the strain were non-motile, halophilic, fermentative, alginolytic, and gas production. A total of 27 traits (motility, growth temperature range, amylase, alginase and lipase productions, and assimilation of 19 carbon compounds) distinguished the strain from the other species in the Rumoiensis clade. The name V. aphrogenes sp. nov. is proposed for this species in the Rumoiensis clade, with CA-1004T as the type strain (JCM 31643T = DSM 103759T).

  5. Isolation and Characterization of Bacteria Colonizing Acartia tonsa Copepod Eggs and Displaying Antagonist Effects against Vibrio anguillarum, Vibrio alginolyticus and Other Pathogenic Strains

    Directory of Open Access Journals (Sweden)

    Mahammed Zidour

    2017-10-01

    Full Text Available Copepods represent a major source of food for many aquatic species of commercial interest for aquaculture such as mysis shrimp and early stages of fishes. For the purpose of this study, the culturable mesophilic bacterial flora colonizing Acartia tonsa copepod eggs was isolated and identified. A total of 175 isolates were characterized based on their morphological and biochemical traits. The majority of these isolates (70% were Gram-negative bacteria. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS was used for rapid identification of bacterial isolates. Here, 58% of isolates were successfully identified at the genus level and among them, 54% were identified at the species level. These isolates belong to 12 different genera and 29 species. Five strains, identified as Bacillus pumilus, named 18 COPS, 35A COPS, 35R COPS, 38 COPS, and 40A COPS, showed strong antagonisms against several potential fish pathogens including Vibrio alginolyticus, V. anguillarum, Listeria monocytogenes, and Staphylococcus aureus. Furthermore, using a differential approach, we show that the antimicrobial activity of the 35R COPS strain is linked primarily to the production of antimicrobial compounds of the amicoumacin family, as demonstrated by the specific UV-absorbance and the MS/MS fragmentation patterns of these compounds.

  6. Antimicrobial effect of dietary oregano essential oil against Vibrio bacteria in shrimps

    OpenAIRE

    Gracia-Valenzuela M.H.; Vergara-Jiménez M.J.; Baez-Flores M.E.; Cabrera-Chavez F.

    2014-01-01

    The effect of dietary oregano essential oils on the growth of Vibrio bacteria in shrimps was evaluated. Shrimps were fed: (i) food with oregano oil with a high level of thymol; (ii) food with oregano oil with a high level of carvacrol, and (iii) food without oregano oil (the control). The animals were infected by three species of Vibrio (vulnificus, parahaemolyticus and cholerae). The microbial counts of Vibrio species were significantly lower (p

  7. Complete genome sequence for the shellfish pathogen Vibrio coralliilyticus RE98 isolated from a shellfish hatchery

    Science.gov (United States)

    Vibrio coralliilyticus is a pathogen of corals and larval shellfish. Publications on strain RE98 list it as a Vibrio tubiashii; however, whole genome sequencing confirms RE98 as V. coralliilyticus containing a total of 6,037,824 bp consisting of two chromosomes (3,420,228 and 1,917,482 bp), and two...

  8. Isolation, characterization, and antibiotic resistance of Vibrio spp. in sea turtles from Northwestern Mexico

    Science.gov (United States)

    Zavala-Norzagaray, Alan A.; Aguirre, A. Alonso; Velazquez-Roman, Jorge; Flores-Villaseñor, Héctor; León-Sicairos, Nidia; Ley-Quiñonez, C. P.; Hernández-Díaz, Lucio De Jesús; Canizalez-Roman, Adrian

    2015-01-01

    The aerobic oral and cloacal bacterial microbiota and their antimicrobial resistance were characterized for 64 apparently healthy sea turtles captured at their foraging grounds in Ojo de Liebre Lagoon (OLL), Baja California Sur (BCS), Mexico (Pacific Ocean) and the lagoon system of Navachiste (LSN) and Marine Area of Influence (MAI), Guasave, Sinaloa (Gulf of California). A total of 34 black turtles (Chelonia mydas agassizii) were sampled in OLL and eight black turtles and 22 olive ridley turtles (Lepidochelys olivacea) were sampled in LSN and MAI, respectively from January to December 2012. We isolated 13 different species of Gram-negative bacteria. The most frequently isolated bacteria were Vibrio alginolyticus in 39/64 (60%), V. parahaemolyticus in 17/64 (26%), and V. cholerae in 6/64 (9%). However, V. cholerae was isolated only from turtles captured from the Gulf of California (MAI). Among V. parahaemolyticus strains, six O serogroups and eight serovars were identified from which 5/17 (29.4%) belonged to the pathogenic strains (tdh+ gene) and 2/17 (11.7%) had the pandemic clone (tdh+ and toxRS/new+). Among V. cholerae strains, all were identified as non-O1/non-O139, and in 4/6 (66%) the accessory cholera enterotoxin gene (ace) was identified but without virulence gene zot, ctxA, and ctxB. Of the isolated V. parahaemolyticus, V. cholerae, and V. alginolyticus strains, 94.1, 33.4, and 100% demonstrated resistance to at least one commonly prescribed antibiotic (primarily to ampicillin), respectively. In conclusion, the presence of several potential (toxigenic) human pathogens in sea turtles may represent transmission of environmental microbes and a high-risk of food-borne disease. Therefore, based on the fact that it is illegal and unhealthy, we discourage the consumption of sea turtle meat or eggs in northwestern Mexico. PMID:26161078

  9. Isolation, characterization, and antibiotic resistance of Vibrio spp. in sea turtles from Northwestern Mexico.

    Science.gov (United States)

    Zavala-Norzagaray, Alan A; Aguirre, A Alonso; Velazquez-Roman, Jorge; Flores-Villaseñor, Héctor; León-Sicairos, Nidia; Ley-Quiñonez, C P; Hernández-Díaz, Lucio De Jesús; Canizalez-Roman, Adrian

    2015-01-01

    The aerobic oral and cloacal bacterial microbiota and their antimicrobial resistance were characterized for 64 apparently healthy sea turtles captured at their foraging grounds in Ojo de Liebre Lagoon (OLL), Baja California Sur (BCS), Mexico (Pacific Ocean) and the lagoon system of Navachiste (LSN) and Marine Area of Influence (MAI), Guasave, Sinaloa (Gulf of California). A total of 34 black turtles (Chelonia mydas agassizii) were sampled in OLL and eight black turtles and 22 olive ridley turtles (Lepidochelys olivacea) were sampled in LSN and MAI, respectively from January to December 2012. We isolated 13 different species of Gram-negative bacteria. The most frequently isolated bacteria were Vibrio alginolyticus in 39/64 (60%), V. parahaemolyticus in 17/64 (26%), and V. cholerae in 6/64 (9%). However, V. cholerae was isolated only from turtles captured from the Gulf of California (MAI). Among V. parahaemolyticus strains, six O serogroups and eight serovars were identified from which 5/17 (29.4%) belonged to the pathogenic strains (tdh (+) gene) and 2/17 (11.7%) had the pandemic clone (tdh (+) and toxRS/new (+)). Among V. cholerae strains, all were identified as non-O1/non-O139, and in 4/6 (66%) the accessory cholera enterotoxin gene (ace) was identified but without virulence gene zot, ctxA, and ctxB. Of the isolated V. parahaemolyticus, V. cholerae, and V. alginolyticus strains, 94.1, 33.4, and 100% demonstrated resistance to at least one commonly prescribed antibiotic (primarily to ampicillin), respectively. In conclusion, the presence of several potential (toxigenic) human pathogens in sea turtles may represent transmission of environmental microbes and a high-risk of food-borne disease. Therefore, based on the fact that it is illegal and unhealthy, we discourage the consumption of sea turtle meat or eggs in northwestern Mexico.

  10. Isolation, Characterization, and Antibiotic Resistance of Vibrio spp. in Sea Turtles from Northwestern Mexico

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    Alan A. eZavala-Norzagaray

    2015-06-01

    Full Text Available The aerobic oral and cloacal bacterial microbiota and their antimicrobial resistance were characterized for 64 apparently healthy sea turtles captured at their foraging grounds in Ojo de Liebre Lagoon (OLL, Baja California Sur, Mexico (Pacific Ocean and the lagoon system of Navachiste (LSN and Marine Area of Influence (MAI, Guasave, Sinaloa (Gulf of California. A total of 34 black turtles (Chelonia mydas agassizii were sampled in OLL and eight black turtles and 22 olive ridley turtles (Lepidochelys olivacea were sampled in LSN and MAI, respectively from January to December 2012. We isolated 13 different species of Gram-negative bacteria. The most frequently isolated bacteria were Vibrio alginolyticus in 39/64 (60%, V. parahaemolyticus in 17/64 (26% and V. cholerae in 6/64 (9%,. However, V. cholerae was isolated only from turtles captured from the Gulf of California (MAI. Among V. parahaemolyticus strains, six O serogroups and eight serovars were identified from which 5/17 (29.4% belonged to the pathogenic strains (tdh+ gene and 2/17 (11.7% had the pandemic clone (tdh+ and toxRS/new+. Among V. cholerae strains, all were identified as non-O1/non-O139, and in 4/6 (66% the accessory cholera enterotoxin gene (ace was identified but without virulence gene zot, ctxA and ctxB. Of the isolated V. parahaemolyticus, V. cholerae and V. alginolyticus strains, 94.1%, 33.4% and 100% demonstrated resistance to at least one commonly prescribed antibiotic (primarily to ampicillin, respectively. In conclusion, the presence of several potential (toxigenic human pathogens in sea turtles may represent transmission of environmental microbes and a high-risk of food-borne disease. Therefore, based on the fact that it is illegal and unhealthy, we discourage the consumption of sea turtle meat or eggs in northwestern Mexico.

  11. Prevalence and Antimicrobial Susceptibility of Vibrio parahaemolyticus Isolated from Short Mackerels (Rastrelliger brachysoma in Malaysia

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    Chia W. Tan

    2017-06-01

    Full Text Available Numerous prevalence studies and outbreaks of Vibrio parahaemolyticus infection have been extensively reported in shellfish and crustaceans. Information on the quantitative detection of V. parahaemolyticus in finfish species is limited. In this study, short mackerels (Rastrelliger brachysoma obtained from different retail marketplaces were monitored with the presence of total and pathogenic strains of V. parahaemolyticus. Out of 130 short mackerel samples, 116 (89.2% were detected with the presence of total V. parahaemolyticus and microbial loads of total V. parahaemolyticus ranging from <3 to >105 MPN/g. Prevalence of total V. parahaemolyticus was found highest in wet markets (95.2% followed by minimarkets (89.1% and hypermarkets (83.3%. Pathogenic V. parahaemolyticus strains (tdh+ and/or trh+ were detected in 16.2% (21 of 130 of short mackerel samples. The density of tdh+ V. parahaemolyticus strains were examined ranging from 3.6 to >105 MPN/g and microbial loads of V. parahaemolyticus strains positive for both tdh and trh were found ranging from 300 to 740 MPN/g. On the other hand, antibiotic susceptibility profiles of V. parahaemolyticus strains isolated from short mackerels were determined through disc diffusion method in this study. Assessment of antimicrobial susceptibility profile of V. parahaemolyticus revealed majority of the isolates were highly susceptible to ampicillin sulbactam, meropenem, ceftazidime, and imipenem, but resistant to penicillin G and ampicillin. Two isolates (2.99% exhibited the highest multiple antibiotic resistance (MAR index value of 0.41 which shown resistance to 7 antibiotics. Results of the present study demonstrated that the occurrence of pathogenic V. parahaemolyticus strains in short mackerels and multidrug resistance of V. parahaemolyticus isolates could be a potential public health concerns to the consumer. Furthermore, prevalence data attained from the current study can be further used to develop a

  12. Genome Sequence of Vibrio campbellii Strain UMTGB204, a Marine Bacterium Isolated from a Green Barrel Tunicate

    Science.gov (United States)

    Gan, Huan You; Noor, Mohd Ezhar Mohd; Saari, Nur Azna; Musa, Najiah; Mustapha, Baharim; Usup, Gires

    2015-01-01

    Vibrio campbellii strain UMTGB204 was isolated from a green barrel tunicate. The genome of this strain comprises 5,652,224 bp with 5,014 open reading frames, 9 rRNAs, and 116 tRNAs. It contains genes related to virulence and environmental tolerance. Gene clusters for the biosynthesis of nonribosomal peptides and bacteriocin were also identified. PMID:25814609

  13. New Sequence Types of Vibrio parahaemolyticus Isolated from a Malaysian Aquaculture Pond, as Revealed by Whole-Genome Sequencing.

    Science.gov (United States)

    Foo, Soon Man; Eng, Wilhelm Wei Han; Lee, Yin Peng; Gui, Kimberly; Gan, Han Ming

    2017-05-11

    The acquisition of Photorhabdus insect-related (Pir) toxin-like genes in Vibrio parahaemolyticus has been linked to hepatopancreatic necrosis disease in shrimp. We report the whole-genome sequences of genetically virulent and avirulent V. parahaemolyticus isolated from a Malaysian aquaculture pond and show that they represent previously unreported sequence types of V. parahaemolyticus. Copyright © 2017 Foo et al.

  14. Prevalence, characterization, and antibiotic susceptibility of Vibrio parahaemolyticus isolated from retail aquatic products in North China.

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    Xu, Xiaoke; Cheng, Jianheng; Wu, Qingping; Zhang, Jumei; Xie, Tengfei

    2016-03-09

    Vibrio parahaemolyticus is a major foodborne pathogen, particularly in Asian countries. Increased occurrence of outbreaks of V. parahaemolyticus gastroenteritis in China indicates the need to evaluation of the prevalence of this pathogenic species. V. parahaemolyticus distribution in shellfish from the eastern coast of China has been reported previously. However, to date, the prevalence of V. parahaemolyticus in retail aquatic products in North China has not been determined. To investigate the prevalence of V. parahaemolyticus in aquatic products in North China, 260 aquatic product samples were obtained from retail markets in 6 provinces of North China from November to December in 2012 and July to August in 2013. V. parahaemolyticus was detected in 94 (36.2%) of the samples by the most probable number method. The density of V. parahaemolyticus ranged from 1.50 to 1100 MPN/g. V. parahaemolyticus was detected at a rate of 50.0% and 22.7% in summer and in winter, respectively. The density of V. parahaemolyticus was significantly higher in summer than in winter, with mean levels of 16.5 MPN/g and 5.0 MPN/g, respectively. Among 145 V. parahaemolyticus isolates examined, none of the isolates possessed tdh and trh. In multiplex PCR-based O-antigen serotyping of these 145 isolates, all serotypes, other than O6, O7, and O9, were detected, and serotype O2 was found to be the most prevalent (detected in 54 isolates). The 145 isolates were grouped into 7 clusters by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) at a similarity coefficient of 0.66. The antimicrobial resistance patterns of these 145 isolates to 12 antimicrobial agents revealed that most of the isolates were resistant to streptomycin (86.2%), while fewer were resistant to ampicillin (49.6%), cefazolin (43.5%), cephalothin (35.9%), and kanamycin (22.1%). All of the examined isolates were susceptible to azithromycin and chloramphenicol. The findings of this study will help

  15. Isolation and characterization of agar-digesting Vibrio species from the rotten thallus of Gracilariopsis heteroclada Zhang et Xia.

    Science.gov (United States)

    Martinez, Joval N; Padilla, Philip Ian P

    2016-08-01

    Gracilariopsis heteroclada Zhang et Xia (Gracilariaceae, Rhodophyta) is one of the most studied marine seaweeds due to its economic importance. This has been cultivated extensively on commercial scale in the Philippines and other Asian countries. However, sustainable production of G. heteroclada in the Philippines could not be maximized due to the occurrence of rotten thallus disease. Thus, isolation and characterization of agar-digesting bacteria from the rotten thalli of G. heteroclada was conducted. A total of seven representative bacterial isolates were randomly selected based on their ability to digest agar as evidenced by the formation of depressions around the bacterial colonies on nutrient agar plates supplemented with 1.5% NaCl and liquefaction of agar. Gram-staining and biochemical characterization revealed that isolates tested were gram-negative rods and taxonomically identified as Vibrio parahaemolyticus (86-99.5%) and Vibrio alginolyticus (94.2-97.7%), respectively. It is yet to be confirmed whether these agar-digesting vibrios are involved in the induction and development of rotten thallus disease in G. heteroclada in concomitance with other opportunistic bacterial pathogens coupled with adverse environmental conditions. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. The inhibition and resistance mechanisms of actinonin, isolated from marine Streptomyces sp. NHF165, against Vibrio anguillarum

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    Na Yang

    2016-09-01

    Full Text Available Vibrio sp. is the most serious pathogen in marine aquaculture, and the development of anti-Vibrio agents is urgently needed. However, it is extreme lack of high-throughput screening (HTS model for searching anti-Vibrio compounds. Here, we established a protein-based HTS screening model to identify agents targeting peptide deformylase (PDF of Vibrio anguillarum. To find potential anti-Vibrio compounds, crude extracts derived from marine actinomycetes were applied for screening with this model. Notably, crude extract of strain Streptomyces sp. NHF165 inhibited dramatically both on V. anguillarum PDF (VaPDF activity and V. anguillarum cell growth. And actinonin was further identified as the functional component. Anti-VaPDF and anti-V. anguillarum activities of actinonin were dose-dependent, and the IC50 values were 6.94 M and 2.85 M, respectively. To understand the resistance of V. anguillarum against actinonin, spontaneous V. anguillarum mutants with resistance against actinonin were isolated. Surprisingly, for the resistant strains, the region between 774 and 852 base pairs was found to be absent in the gene folD which produces 10-formyl-tetrahydrofolate, a donor of N-formyl to Met-tRNAfmet. When compared to the wild type strain, folD mutant showed 8 times of minimum inhibition concentration on actinonin, however, the folD complementary strain could not grow on the medium supplemented with actinonin, which suggested that folD gene mutation was mainly responsible for the actinonin resistance. To our knowledge, this is the first report showing that marine derived Streptomyces sp. could produce actinonin with anti-VaPDF activity and the resistance against actinonin by V. anguillarum is mediated by mutation in folD gene.

  17. Genetic diversity of environmental Vibrio cholerae O1 strains isolated in Northern Vietnam.

    Science.gov (United States)

    Takemura, Taichiro; Murase, Kazunori; Maruyama, Fumito; Tran, Thi Luong; Ota, Atsushi; Nakagawa, Ichiro; Nguyen, Dong Tu; Ngo, Tu Cuong; Nguyen, Thi Hang; Tokizawa, Asako; Morita, Masatomo; Ohnishi, Makoto; Nguyen, Binh Minh; Yamashiro, Tetsu

    2017-10-01

    Cholera epidemics have been recorded periodically in Vietnam during the seventh cholera pandemic. Since cholera is a water-borne disease, systematic monitoring of environmental waters for Vibrio cholerae presence is important for predicting and preventing cholera epidemics. We conducted monitoring, isolation, and genetic characterization of V. cholerae strains in Nam Dinh province of Northern Vietnam from Jul 2013 to Feb 2015. In this study, four V. cholerae O1 strains were detected and isolated from 110 analyzed water samples (3.6%); however, none of them carried the cholera toxin gene, ctxA, in their genomes. Whole genome sequencing and phylogenetic analysis revealed that the four O1 isolates were separated into two independent clusters, and one of them diverged from a common ancestor with pandemic strains. The analysis of pathogenicity islands (CTX prophage, VPI-I, VPI-II, VSP-I, and VSP-II) indicated that one strain (VNND_2014Jun_6SS) harbored an unknown prophage-like sequence with high homology to vibriophage KSF-1 phi and VCY phi, identified from Bangladesh and the USA, respectively, while the other three strains carried tcpA gene with a distinct sequence demonstrating a separate clonal lineage. These results suggest that the aquatic environment can harbor highly divergent V. cholera strains and serve as a reservoir for multiple V. cholerae virulence-associated genes which may be exchanged via mobile genetic elements. Therefore, continuous monitoring and genetic characterization of V. cholerae strains in the environment should contribute to the early detection of the sources of infection and prevention of cholera outbreaks as well as to understanding the natural ecology and evolution of V. cholerae. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  18. Isolation and characterization of specific bacteriophage Va1 to Vibrio alginolyticus

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    Carla Fernández Espinel

    2017-04-01

    Full Text Available Vibrio alginolyticus is associated with diseases in aquaculture. The misuse of antibiotics has led to the search for alternatives in the treatment of bacterial diseases, among them the application of bacteriophages that infect and destroy bacteria selectively. In this way, a highly lytic V. alginolyticus bacteriophage, termed Va1, was isolated, with the aim to evaluate its physical chemical parameters. For this purpose, different temperature, pH, chloroform exposure and host range conditions were evaluated. The temperature stability of phage Va1 showed higher titers at 20 and 30 °C decreasing from 40 °C. With respect to pH, the highest titers for the bacteriophage were between 5 and 8, and chloroform exposure reduced viability of the Va1 phage by 25%. The one-step curve determined that the latency period and the burst size were 20 minutes and 192 PFU / infective center respectively. Under the transmission electron microscope, the Va1 phage showed an icosahedral head and a non-contractile tail, belonging to the Podoviridae family. In conclusion, Va1 phage presents potential characteristics for use in phage therapy.

  19. Antibiotic resistance of Vibrio parahaemolyticus isolated from pond-reared Litopenaeus vannamei marketed in Natal, Brazil

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    Ligia Maria Rodrigues de Melo

    2011-12-01

    Full Text Available Ten out of fifty fresh and refrigerated samples of shrimp (Litopenaeus vannamei collected from retailers in Natal (Rio Grande do Norte, Northeastern Brazil tested positive for Vibrio parahaemolyticus. The Kanagawa test and multiplex PCR assays were used to detect TDH and TRH hemolysins and the tdh, trh and tlh genes, respectively. All strains were Kanagawa-negative and tlh-positive. Antibiotic susceptibility testing was done for seven antibiotics by the agar diffusion technique. Five strains (50% presented multiple antibiotic resistance to ampicillin (90% and amikacin (60%, while two strains (20% displayed intermediate-level resistance to amikacin. All strains were sensitive to chloramphenicol. Intermediate-level susceptibility and/or resistance to other antibiotics ranged from 10 to 90%, with emphasis on the observed growing intermediate-level resistance to ciprofloxacin. Half our isolates yielded a multiple antibiotic resistance index above 0.2 (range: 0.14-0.29, indicating a considerable risk of propagation of antibiotic resistance throughout the food chain.

  20. Antibiotic resistance of Vibrio parahaemolyticus isolated from pond-reared Litopenaeus vannamei marketed in natal, brazil

    Science.gov (United States)

    de Melo, Ligia Maria Rodrigues; Almeida, Dulce; Hofer, Ernesto; dos Reis, Cristhiane Moura Falavina; Theophilo, Grace Nazareth Diogo; Santos, André Felipe das Mercês; Vieira, Regine Helena Silva dos Fernandes

    2011-01-01

    Ten out of fifty fresh and refrigerated samples of shrimp (Litopenaeus vannamei) collected from retailers in Natal (Rio Grande do Norte, Northeastern Brazil) tested positive for Vibrio parahaemolyticus. The Kanagawa test and multiplex PCR assays were used to detect TDH and TRH hemolysins and the tdh, trh and tlh genes, respectively. All strains were Kanagawa-negative and tlh-positive. Antibiotic susceptibility testing was done for seven antibiotics by the agar diffusion technique. Five strains (50%) presented multiple antibiotic resistance to ampicillin (90%) and amikacin (60%), while two strains (20%) displayed intermediate-level resistance to amikacin. All strains were sensitive to chloramphenicol. Intermediate-level susceptibility and/or resistance to other antibiotics ranged from 10 to 90%, with emphasis on the observed growing intermediate-level resistance to ciprofloxacin. Half our isolates yielded a multiple antibiotic resistance index above 0.2 (range: 0.14–0.29), indicating a considerable risk of propagation of antibiotic resistance throughout the food chain. PMID:24031779

  1. [Comparative genomic analysis of vibrio cholerae El Tor preseventh and seventh pandemic strains isolated in various periods].

    Science.gov (United States)

    Osin, A V; Nefedov, K S; Eroshenko, G A; Smirnova, N I

    2005-01-01

    Genetic organization of 52 Vibrio cholerae El Tor biotype preseventh and seventh pandemic strains isolated in various periods was studied by PCR assay and DNA-DNA hybridization. It was established that the genome of most ancient of analyzed strains isolated from a diarrhea patient in 1910 was devoid of CTX and RS1 prophages, vibrio pathogenicity islands (VPI and VPI-2), and pandemic islands (VSP-1 and VSP-2) that contain key virulence genes. The appearance of pathogenic properties in cholera vibrios for the first time causing a local outbreak of cholera in 1937 is connected with the acquisition of VPI and CTX that carried genes tcpA and ctx-AB, respectively, which are responsible for the colonization of small intestine and encode the production of cholera toxin. The appearance of seventh pandemic agent for cholera was shown to correlate with the acquisition by its precursor of two additional blocks of genes VSP-1 and VSP-2. This finding strongly supports the involvement of these genes in formation of the pandemic potential in strains. Molecular typing methods allowed elucidation of differences in the genetic organization between prepandemic and pandemic strains. The detected variability of the genome of contemporary virulent strains may be a reason for the occurrence of etiological agent for cholera with new properties.

  2. Molecular Typing of Vibrio parahaemolyticus Strains Isolated from Mollusks in the North Adriatic Sea.

    Science.gov (United States)

    Rahman, Mohammad Shamsur; Carraro, Roberta; Cardazzo, Barbara; Carraro, Lisa; Meneguolo, Davide Boscolo; Martino, Maria Elena; Andreani, Nadia Andrea; Bordin, Paola; Mioni, Renzo; Barco, Lisa; Novelli, Enrico; Balzan, Stefania; Fasolato, Luca

    2017-08-01

    Vibrio parahaemolyticus is an emerging foodborne pathogen in the Mediterranean, usually associated with shellfish consumption. The increase in the number of outbreaks in Europe is primarily associated with the global warming of the ocean that has a great impact on the spread and genetic selection of waterborne pathogens. The primary role of Italy in Europe's mollusk production, together with the fact that cases of infections with V. parahaemolyticus are not always notified to the European community, highlighted the necessity of acquiring new information about the epidemiological involvement of shellfish products. The aim of the study was to provide useful insights into the first steps of the Risk Assessment associated with V. parahaemolyticus through the molecular characterization of isolates from commercialized mollusks. A total of 102 strains identified as V. parahaemolyticus were investigated as part of a larger sampling (1-year survey) from several shellfish species collected from the Venice lagoon and the North Adriatic sea. All strains were characterized by multilocus sequence typing and tested for the presence of virulence genes (trh and tdh). The study of sampling/environmental factors and epidemiological analyses was performed to describe the behaviors of the different genetic populations. The population structure analysis highlighted three genetic clusters that could be subject to temperature selection during cold (≤15°C) and warm (>16°C) seasons. Moreover, other factors, such as molluscan species (clams/mussels), probably played a role in the distribution of genetic clusters. Although few strains carried the virulence factors (n = 6 trh+), epidemiological links with clinical isolates and a local dissemination of some sequence types were underlined. This work provides a useful background on the genotype spread as a first step in the Hazard Identification in light of future climate changes.

  3. Multiple Antibiotic Resistances of Vibrio Isolates from Coastal and Brackish Water Areas

    OpenAIRE

    S. Manjusha; G. B. Sarita; K. K. Elyas; Chandrasekaran, M.

    2005-01-01

    An experiment was designed to assess the occurrence of multiple antibiotic resistances in Vibrio spp. from different (brackish and marine) environments. Water samples from nine marine landing sites and two coastal inland aquaculture farms were screened for the Vibrio spp. and assessed their resistance to twenty-two different antibiotics, which are commonly encountered in the aquatic ecosystem. Tissue samples (shrimp, mussel and sepia) were tested from the sampling site wit...

  4. Biosurfactant production by hydrocarbon-degrading Brevibacterium and Vibrio isolates from the sea pen Pteroeides spinosum (Ellis, 1764).

    Science.gov (United States)

    Graziano, Marco; Rizzo, Carmen; Michaud, Luigi; Porporato, Erika Maria Diletta; De Domenico, Emilio; Spanò, Nunziacarla; Lo Giudice, Angelina

    2016-09-01

    Among filter-feeders, pennatulids are the most complex and polymorphic members of the cnidarian class Anthozoa. They display a wide distribution throughout all the oceans, constituting a significant component of the sessile megafauna from intertidal to abyssal depths. In this study, a total of 118 bacterial isolates from enrichment cultures, carried out with homogenates of the sea pen Pteroeides spinosum (Ellis, 1764), were screened for hydrocarbon utilization by using the 2,6-dichlorophenol indophenol assay. Among them, 83 hydrocarbon-oxidizing isolates were analyzed for biosurfactant production by standard screening tests (i.e., emulsifying activity, E24 detection, surface tension measurement, microplate assay). The 16S rRNA gene sequencing revealed the affiliation of the most promising isolates to the genera Brevibacterium and Vibrio. Biosurfactant production resulted strongly affected by salinity and temperature conditions, and occurred in the presence of diesel oil and/or crude oil, whereas no production was observed when isolates were grown on tetradecane. The strains resulted able to create stable emulsions, thus suggesting the production of biosurfactants. Further analyses revealed a glycolipidic nature of the biosurfactant extracted from Vibrio sp. PBN295, a genus that has been only recently reported as biosurfactant producer. Results suggest that pennatulids could represent a novel source for the isolation of hydrocarbon-oxidizing bacteria with potential in biosurfactant production. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Characterization of trh2 Harbouring Vibrio parahaemolyticus Strains Isolated in Germany

    Science.gov (United States)

    Bechlars, Silke; Jäckel, Claudia; Diescher, Susanne; Wüstenhagen, Doreen A.; Kubick, Stefan; Dieckmann, Ralf; Strauch, Eckhard

    2015-01-01

    Background Vibrio parahaemolyticus is a recognized human enteropathogen. Thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) as well as the type III secretion system 2 (T3SS2) are considered as major virulence factors. As tdh positive strains are not detected in coastal waters of Germany, we focused on the characterization of trh positive strains, which were isolated from mussels, seawater and patients in Germany. Results Ten trh harbouring V. parahaemolyticus strains from Germany were compared to twenty-one trh positive strains from other countries. The complete trh sequences revealed clustering into three different types: trh1 and trh2 genes and a pseudogene Ψtrh. All German isolates possessed alleles of the trh2 gene. MLST analysis indicated a close relationship to Norwegian isolates suggesting that these strains belong to the autochthonous microflora of Northern Europe seawaters. Strains carrying the pseudogene Ψtrh were negative for T3SS2β effector vopC. Transcription of trh and vopC genes was analyzed under different growth conditions. Trh2 gene expression was not altered by bile while trh1 genes were inducible. VopC could be induced by urea in trh2 bearing strains. Most trh1 carrying strains were hemolytic against sheep erythrocytes while all trh2 positive strains did not show any hemolytic activity. TRH variants were synthesized in a prokaryotic cell-free system and their hemolytic activity was analyzed. TRH1 was active against sheep erythrocytes while TRH2 variants were not active at all. Conclusion Our study reveals a high diversity among trh positive V. parahaemolyticus strains. The function of TRH2 hemolysins and the role of the pseudogene Ψtrh as pathogenicity factors are questionable. To assess the pathogenic potential of V. parahaemolyticus strains a differentiation of trh variants and the detection of T3SS2β components like vopC would improve the V. parahaemolyticus diagnostics and could lead to a refinement of the risk

  6. Detection of virulence associated genes, haemolysin and protease amongst Vibrio cholerae isolated in Malaysia.

    Science.gov (United States)

    Iyer, L; Vadivelu, J; Puthucheary, S D

    2000-08-01

    Eighty-four strains of Vibrio cholerae O1, O139 and non-O1/non-O139 from clinical and environmental sources were investigated for the presence of the toxin co-regulated pilus gene, tcpA, the virulence cassette genes ctxA, zot, ace and cep and also for their ability to elaborate haemolysin and protease. The ctxA and zot genes were detected using DNA-DNA hybridization while the ace, cep and tcpA genes were detected using PCR. Production of haemolysin and protease was detected using mammalian erythrocytes and an agar diffusion assay respectively. Analysis of their virulence profiles showed six different groups designated Type I to Type VI and the major distinguishing factor among these profiles was in the in vitro production of haemolysin and/or protease. Clinical O1, O139 and environmental O1 strains were similar with regard to presence of the virulence cassette genes. All environmental O1 strains with the exception of one were found to possess ctxA, zot and ace giving rise to the probability that these strains may actually be of clinical origin. One strain which had only cep but none of the toxin genes may be a true environmental isolate. The virulence cassette and colonization factor genes were absent in all non-O1/non-O139 environmental strains but production of both the haemolysin and protease was present, indicating that these may be putative virulence factors. These findings suggest that with regard to its pathogenic potential, only strains of the O1 and O139 serogroup that possess the tcpA gene which encodes the phage receptor, have the potential to acquire the CTX genetic element and become choleragenic.

  7. Molecular and Genomic Characterization of Vibrio mimicus Isolated from a Frozen Shrimp Processing Facility in Mexico.

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    Iliana Guardiola-Avila

    Full Text Available Vibrio mimicus is a gram-negative bacterium responsible for diseases in humans. Three strains of V. mimicus identified as V. mimicus 87, V. mimicus 92 and V. mimicus 93 were isolated from a shrimp processing facility in Guaymas, Sonora, Mexico. The strains were analyzed using several molecular techniques and according to the cluster analysis they were different, their similarities ranged between 51.3% and 71.6%. ERIC-PCR and RAPD (vmh390R were the most discriminatory molecular techniques for the differentiation of these strains. The complete genomes of two strains (V. mimicus 87, renamed as CAIM 1882, and V. mimicus 92, renamed as CAIM 1883 were sequenced. The sizes of the genomes were 3.9 Mb in both strains, with 2.8 Mb in ChI and 1.1 Mb in ChII. A 12.7% difference was found in the proteome content (BLAST matrix. Several virulence genes were detected (e.g. capsular polysaccharide, an accessory colonization factor and genes involved in quorum-sensing which were classified in 16 categories. Variations in the gene content between these genomes were observed, mainly in proteins and virulence genes (e.g., hemagglutinin, mobile elements and membrane proteins. According to these results, both strains were different, even when they came from the same source, giving an insight of the diversity of V. mimicus. The identification of various virulence genes, including a not previously reported V. mimicus gene (acfD in ChI in all sequenced strains, supports the pathogenic potential of this species. Further analysis will help to fully understand their potential virulence, environmental impact and evolution.

  8. Exploring the Genomic Traits of Non-toxigenic Vibrio parahaemolyticus Strains Isolated in Southern Chile

    Directory of Open Access Journals (Sweden)

    Daniel Castillo

    2018-02-01

    Full Text Available Vibrio parahaemolyticus is the leading cause of seafood-borne gastroenteritis worldwide. As reported in other countries, after the rise and fall of the pandemic strain in Chile, other post-pandemic strains have been associated with clinical cases, including strains lacking the major toxins TDH and TRH. Since the presence or absence of tdh and trh genes has been used for diagnostic purposes and as a proxy of the virulence of V. parahaemolyticus isolates, the understanding of virulence in V. parahaemolyticus strains lacking toxins is essential to detect these strains present in water and marine products to avoid possible food-borne infection. In this study, we characterized the genome of four environmental and two clinical non-toxigenic strains (tdh-, trh-, and T3SS2-. Using whole-genome sequencing, phylogenetic, and comparative genome analysis, we identified the core and pan-genome of V. parahaemolyticus of strains of southern Chile. The phylogenetic tree based on the core genome showed low genetic diversity but the analysis of the pan-genome revealed that all strains harbored genomic islands carrying diverse virulence and fitness factors or prophage-like elements that encode toxins like Zot and RTX. Interestingly, the three strains carrying Zot-like toxin have a different sequence, although the alignment showed some conserved areas with the zot sequence found in V. cholerae. In addition, we identified an unexpected diversity in the genetic architecture of the T3SS1 gene cluster and the presence of the T3SS2 gene cluster in a non-pandemic environmental strain. Our study sheds light on the diversity of V. parahaemolyticus strains from the southern Pacific which increases our current knowledge regarding the global diversity of this organism.

  9. Biodegradable and biocompatible biomaterial, polyhydroxybutyrate, produced by an indigenous Vibrio sp. BM-1 isolated from marine environment.

    Science.gov (United States)

    Wei, Yu-Hong; Chen, Wei-Chuan; Wu, Ho-Shing; Janarthanan, Om-Murugan

    2011-01-01

    Polyhydroxybutyrate (PHB) is one of the polyhydroxyalkanoates (PHAs) which has biodegradable and biocompatible properties. They are adopted in the biomedical field, in, for example, medical implants and drug delivery carriers. This study seeks to promote the production of PHB by Vibrio sp. BM-1, isolated from a marine environment by improving constituents of medium and implementing an appropriate fermentation strategy. This study successfully developed a glycerol-yeast extract-tryptone (GYT) medium that can facilitate the growth of Vibrio sp. BM-1 and lead to the production of 1.4 g/L PHB at 20 h cultivation. This study also shows that 1.57 g/L PHB concentration and 16% PHB content were achieved, respectively, when Vibrio sp. BM-1 was cultivated with MS-GYT medium (mineral salts-supplemented GYT medium) for 12 h. Both cell dry weight (CDW) and residual CDW remained constant at around 8.2 g/L and 8.0 g/L after the 12 h of cultivation, until the end of the experiment. However, both 16% of PHB content and 1.57 g/L of PHB production decreased rapidly to 3% and 0.25 g/L, respectively from 12 h of cultivation to 40 h of cultivation. The results suggest that the secretion of PHB depolymerase that might be caused by the addition of mineral salts reduced PHB after 12 h of cultivation. However, work will be done to explain the effect of adding mineral salts on the production of PHB by Vibrio sp. BM-1 in the near future.

  10. Vibrio cholerae Serogroup O139: Isolation from Cholera Patients and Asymptomatic Household Family Members in Bangladesh between 2013 and 2014.

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    Fahima Chowdhury

    2015-11-01

    Full Text Available Cholera is endemic in Bangladesh, with outbreaks reported annually. Currently, the majority of epidemic cholera reported globally is El Tor biotype Vibrio cholerae isolates of the serogroup O1. However, in Bangladesh, outbreaks attributed to V. cholerae serogroup O139 isolates, which fall within the same phylogenetic lineage as the O1 serogroup isolates, were seen between 1992 and 1993 and in 2002 to 2005. Since then, V. cholerae serogroup O139 has only been sporadically isolated in Bangladesh and is now rarely isolated elsewhere.Here, we present case histories of four cholera patients infected with V. cholerae serogroup O139 in 2013 and 2014 in Bangladesh. We comprehensively typed these isolates using conventional approaches, as well as by whole genome sequencing. Phenotypic typing and PCR confirmed all four isolates belonging to the O139 serogroup.Whole genome sequencing revealed that three of the isolates were phylogenetically closely related to previously sequenced El Tor biotype, pandemic 7, toxigenic V. cholerae O139 isolates originating from Bangladesh and elsewhere. The fourth isolate was a non-toxigenic V. cholerae that, by conventional approaches, typed as O139 serogroup but was genetically divergent from previously sequenced pandemic 7 V. cholerae lineages belonging to the O139 or O1 serogroups.These results suggest that previously observed lineages of V. cholerae O139 persist in Bangladesh and can cause clinical disease and that a novel disease-causing non-toxigenic O139 isolate also occurs.

  11. Antibiotics resistance in El Tor Vibrio cholerae 01 isolated during cholera outbreaks in Mozambique from 2012 to 2015.

    Science.gov (United States)

    Dengo-Baloi, Liliana Candida; Semá-Baltazar, Cynthia Amino; Manhique, Lena Vania; Chitio, Jucunu Elias; Inguane, Dorteia Luísa; Langa, José Paulo

    2017-01-01

    Mozambique has recorded cyclically epidemic outbreaks of cholera. Antibiotic therapy is recommended in specific situations for management and control of cholera outbreaks. However, an increase in resistance rates to antibiotics by Vibrio cholerae has been reported in several epidemic outbreaks worldwide. On the other hand, there are few recent records of continuous surveillance of antibiotics susceptibility pattern of V. cholerae in Mozambique. The purpose of this study was to evaluate antibiotics resistance pattern of Vibrio cholerae O1 Ogawa isolated during Cholera outbreaks in Mozambique to commonly used antibiotics. We analyzed data from samples received in the context of surveillance and response to Cholera outbreaks in the National Reference Laboratory of Microbiology from the National Institute of Health of Mozambique, 159 samples suspected of cholera from cholera treatment centers of, Metangula (09), Memba (01), Tete City (08), Moatize (01), Morrumbala (01) districts, City of Quelimane (01), Lichinga (06) and Nampula (86) districts, from 2012 to 2015. Laboratory culture and standard biochemical tests were employed to isolate and identify Vibrio cholerae; serotypes were determined by antisera agglutination reaction in blade. Biotype and presence of important virulence factors analysis was done by PCR. Antibiotics susceptibility pattern was detected by disk diffusion method Kirby Bauer. Antibiotic susceptibility and results were interpreted by following as per recommendations of CLSI (Clinical and Laboratory Standards Institute) 2014. All samples were collected and tested in the context of Africhol Project, approved by the National Bioethics Committee for Health. Among isolates from of Vibrio cholerae O1 El Tor Ogawa resistance to Sulphamethoxazole-trimethropim was 100% (53/53) to Trimethoprim-, being 100% (54/54) for Ampicillin, 99% (72/74) for Nalidixic Acid, 97% (64/66) to Chloramphenicol, 95% (42/44) for Nitrofurantoin and (19/20) Cotrimoxazole, 83% (80

  12. Molecular Analysis and Toxigenic Potential of Vibrio cholerae Isolated from Hilsha fish (Tenualosa ilisha), Bangladesh

    DEFF Research Database (Denmark)

    Hossain, Zenat Zebin; Farhana, Israt; Tulsiani, Suhella

    Exposure to contaminated fish may upsurge the virulent strains of Vibrio cholerae, the deadly human pathogen in the households of rural and urban Bangladesh. Since V. cholerae spreading was reported from the Bay of Bengal, this study hypothesized that Hilsha (Tenualosa ilisha), a marine and fresh...

  13. Prevalence and Molecular Typing of Vibrio parahaemolyticus (tdh+) isolated from seafood using PCR-based methods

    Science.gov (United States)

    Vibrio parahaemolyticus is a pathogen most frequently implicated in foodborne outbreaks linked to the consumption of seafood in the coastal cities of China. The pathogenicity of environmental V. parahaemolyticus is mostly correlated with the production of thermostable direct hemolysin (TDH). In orde...

  14. Vibrio parahaemolyticus isolates from southeastern Chinese coast are genetically diverse with circulation of clonal complex 3 strains since 2002.

    Science.gov (United States)

    Yu, Ying; Hu, Weizhao; Wu, Beibei; Zhang, Peipei; Chen, Jianshun; Wang, Shuna; Fang, Weihuan

    2011-11-01

    Multilocus sequence typing (MLST) was used to examine the clonal relationship and genetic diversity of 71 Vibrio parahaemolyticus isolates from clinical and seafood-related sources in southeastern Chinese coast between 2002 and 2009. The tested isolates fell into 61 sequence types (STs). Of 17 clinical isolates, 7 belonged to ST3 of the pandemic clonal complex 3, with 3 strains isolated in 2002. Although there was no apparent clonal relationship found between clinical strains and those from seafood-related sources positive with pathogenic markers, there were clonal relationships between clinical strains from this study and those from environmental sources in other parts of China. Phylogenetic analysis showed that strains of 112 STs (61 STs from this study and 51 retrieved from PUBMLST database covering different continents) could be divided into four branches. The vast majority of our isolates and those from other countries were genetically diverse and clustered into two major branches of mixed distribution (of geographic origins and sample sources), whereas five STs representing six isolates split as two minor branches because of divergence of their recA genes, which had 80%-82% nucleotide identity to typical V. parahaemolyticus strains and 73.3%-76.9% identity to the CDS24 of a Vibrio sp. plasmid p23023, indicating that the recA gene might have recombined by lateral gene transfer. This was further supported by a high ratio of recombination to mutation (3.038) for recA. In conclusion, MLST with fully extractable database is a powerful system for analysis of clonal relationship for strains of a particular region in a national or global scale as well as between clinical and environmental or food-related strains.

  15. The ecological coherence of temperature and salinity tolerance interaction and pigmentation in a non-marine vibrio isolated from Salar de Atacama

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    Karem Gallardo

    2016-12-01

    Full Text Available The occurrence of microorganisms from the Vibrio genus in saline lakes from northern Chile had been evidenced using Numerical Taxonomy decades before and, more recently, by phylogenetic analyses of environmental samples and isolates. Most of the knowledge about this genus came from marine isolates and showed temperature and salinity to be integral agents in shaping the niche of the Vibrio populations. The stress tolerance phenotypes of Vibrio sp. Teb5a1 isolated from Salar de Atacama was investigated. It was able to grow without NaCl and tolerated up to 100 g/L of the salt. Furthermore, it grew between 17° and 49°C (optimum 30°C in the absence of NaCl, and the range was expanded into cold temperature (4-49°C in the presence of the salt. Other additional adaptive strategies were observed in response to the osmotic stress: pigment production, identified as the known antibacterial prodigiosin, swimming and swarming motility and synthesis of a polar flagellum. It is possible to infer that environmental congruence might explain the cellular phenotypes observed in Vibrio sp. considering that coupling between temperature and salinity tolerance, the production of antibacterial agents at higher temperatures, flagellation and motility increase the chance of Vibrio sp. to survive in salty environments with high daily temperature swings and UV radiation.

  16. The Ecological Coherence of Temperature and Salinity Tolerance Interaction and Pigmentation in a Non-marine Vibrio Isolated from Salar de Atacama.

    Science.gov (United States)

    Gallardo, Karem; Candia, Jonathan E; Remonsellez, Francisco; Escudero, Lorena V; Demergasso, Cecilia S

    2016-01-01

    The occurrence of microorganisms from the Vibrio genus in saline lakes from northern Chile had been evidenced using Numerical Taxonomy decades before and, more recently, by phylogenetic analyses of environmental samples and isolates. Most of the knowledge about this genus came from marine isolates and showed temperature and salinity to be integral agents in shaping the niche of the Vibrio populations. The stress tolerance phenotypes of Vibrio sp. Teb5a1 isolated from Salar de Atacama was investigated. It was able to grow without NaCl and tolerated up to 100 g/L of the salt. Furthermore, it grew between 17° and 49°C (optimum 30°C) in the absence of NaCl, and the range was expanded into cold temperature (4-49°C) in the presence of the salt. Other additional adaptive strategies were observed in response to the osmotic stress: pigment production, identified as the known antibacterial prodigiosin, swimming and swarming motility and synthesis of a polar flagellum. It is possible to infer that environmental congruence might explain the cellular phenotypes observed in Vibrio sp. considering that coupling between temperature and salinity tolerance, the production of antibacterial agents at higher temperatures, flagellation and motility increase the chance of Vibrio sp. to survive in salty environments with high daily temperature swings and UV radiation.

  17. Molecular, serological, and virulence characteristics of Vibrio parahaemolyticus isolated from environmental, food, and clinical sources in North America and Asia.

    Science.gov (United States)

    DePaola, Angelo; Ulaszek, Jodie; Kaysner, Charles A; Tenge, Bradley J; Nordstrom, Jessica L; Wells, Joy; Puhr, Nancy; Gendel, Steven M

    2003-07-01

    Potential virulence attributes, serotypes, and ribotypes were determined for 178 pathogenic Vibrio parahaemolyticus isolates from clinical, environmental, and food sources on the Pacific, Atlantic, and Gulf Coasts of the United States and from clinical sources in Asia. The food and environmental isolates were generally from oysters, and they were defined as being pathogenic by using DNA probes to detect the presence of the thermostable direct hemolysin (tdh) gene. The clinical isolates from the United States were generally associated with oyster consumption, and most were obtained from outbreaks in Washington, Texas, and New York. Multiplex PCR was used to confirm the species identification and the presence of tdh and to test for the tdh-related hemolysin trh. Most of the environmental, food, and clinical isolates from the United States were positive for tdh, trh, and urease production. Outbreak-associated isolates from Texas, New York, and Asia were predominantly serotype O3:K6 and possessed only tdh. A total of 27 serotypes and 28 ribogroups were identified among the isolates, but the patterns of strain distribution differed between the serotypes and ribogroups. All but one of the O3:K6 isolates from Texas were in a different ribogroup from the O3:K6 isolates from New York or Asia. The O3:K6 serotype was not detected in any of the environmental and food isolates from the United States, and none of the food or environmental isolates belonged to any of the three ribogroups that contained all of the O3:K6 and related clinical isolates. The combination of serotyping and ribotyping showed that the Pacific Coast V. parahaemolyticus population appeared to be distinct from that of either the Atlantic Coast or Gulf Coast. The fact that certain serotypes and ribotypes contained both clinical and environmental isolates while many others contained only environmental isolates implies that certain serotypes or ribotypes are more relevant for human disease.

  18. Incidence of plasmids in marine Vibrio spp. isolated from an oil field in the northwestern Gulf of Mexico

    Energy Technology Data Exchange (ETDEWEB)

    Hada, H.S.; Sizemore, R.K.

    1981-01-01

    Presumptive marine Vibrio spp. were collected from an operational oil field and control site located in the northwest Gulf of Mexico. Of 440 isolates analyzed for the presence of extrachromosomal deoxyribonucleic acid elements or plasmids by using the cleared lysate and agarose gel techniques, 31% showed distinct plasmid bands on agarose gels. A majority of the plasmids detected were estimated to have mollecular masses of 10 x 10/sup 6/ or less. Multiple plasmids were observed in approximately half of the plasmid-containing strains. A number of isolates contained plasmids with similar banding and mobility patterns. The oil field area had noticeably more plasmid-containing strains (35 versus 23% in the control site) and a greater number of plasmids per plasmid-containing strain (an average of 2.5 plasmids, vs 1.5 in the control site). Oil field discharges might have resulted in increased plasmid incidence and diversity.

  19. Distribution and content of class 1 integrons in different Vibrio cholerae O-serotype strains isolated in Thailand

    DEFF Research Database (Denmark)

    Dalsgaard, Anders; Forslund, Anita; Serichantalergs, Oralak

    2000-01-01

    by the strains. Serotype O139 strains did not contain class 1 integrons. However, the appearance and disappearance of the O139 serotype in the coastal city Samutsakorn in 1992 and 1993 were associated with the emergence of a distinct V. cholerae O1 strain which contained the aad-V resistance gene cassette. A 150......-kb self-transmissible plasmid found in three O1 strains isolated in 1982 contained the aadB gene cassette. Surprisingly, several strains harbored two integrons containing different cassettes. Thus, class 1 integrons containing various resistance gene cassettes are distributed among different V......In this study, 176 clinical and environmental Vibrio cholerae strains of different O serotypes isolated in Thailand from 1982 to 1995 were selected and studied for the presence of class 1 integrons, a new group of genetic elements which carry antibiotic resistance genes. Using PCR and DNA...

  20. Leisingera sp. JC1, a Bacterial Isolate from Hawaiian Bobtail Squid Eggs, Produces Indigoidine and Differentially Inhibits Vibrios

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    Samantha M. Gromek

    2016-09-01

    Full Text Available Female members of many cephalopod species house a bacterial consortium that is part of their reproductive system, the accessory nidamental gland (ANG. These bacteria are deposited into eggs that are then laid in the environment where they must develop unprotected from predation, pathogens and fouling. In this study, we characterized the genome and secondary metabolite production of Leisingera sp. JC1, a member of the roseobacter clade (Rhodobacteraceae of Alphaproteobacteria isolated from the jelly coat of eggs from the Hawaiian bobtail squid, Euprymna scolopes. Whole genome sequencing and MLSA analysis revealed that Leisingera sp. JC1 falls within a group of roseobacters associated with squid ANGs. Genome and biochemical analyses revealed the potential for and production of a number of secondary metabolites, including siderophores and acyl-homoserine lactones involved with quorum sensing. The complete biosynthetic gene cluster for the pigment indigoidine was detected in the genome and mass spectrometry confirmed the production of this compound. Furthermore, we investigated the production of indigoidine under co-culture conditions with Vibrio fischeri, the light organ symbiont of E. scolopes, and with other vibrios. Finally, both Leisingera sp. JC1 and secondary metabolite extracts of this strain had differential antimicrobial activity against a number of marine vibrios, suggesting that Leisingera sp. JC1 may play a role in host defense against other marine bacteria either in the eggs and/or ANG. These data also suggest that indigoidine may be partially, but not wholly, responsible for the antimicrobial activity of this squid-associated bacterium. □

  1. Complete Genome Sequence of Vibrio campbellii LMB 29 Isolated from Red Drum with Four Native Megaplasmids

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    Jinxin Liu

    2017-10-01

    Full Text Available Vibrio spp. are the most common pathogens for animals reared in aquaculture. Vibrio campbellii, which is often involved in shrimp, fish and mollusks diseases, is widely distributed in the marine environment worldwide, but our knowledge about its pathogenesis and antimicrobial resistance is very limited. The existence of this knowledge gap is at least partially because that V. campbellii was originally classified as Vibrio harveyi, and the detailed information of its comparative genome analysis to other Vibrio spp. is currently lacking. In this study, the complete genome of a V. campbellii predominant strain, LMB29, was determined by MiSeq in conjunction with PacBio SMRT sequencing. This genome consists of two circular DNA chromosomes and four megaplasmids. Comparative genome analysis indicates that LMB29 shares a 96.66% similarity (average nucleotide identity with the V. campbellii ATCC strain BAA-1116 based on a 75% AF (average fraction calculations, and its functional profile is very similar to V. campbellii E1 and V. campbellii CAIM115. Both type III secretion system (T3SS and type VI secretion system (T6SS, along with the tlh gene which encodes a thermolabile hemolysin, are present in LMB29 which may contribute to the bacterial pathogenesis. The virulence of this strain was experimental confirmed by performing a LDH assay on a fish cell infection model, and cell death was observed as early as within 3 h post infection. Thirty-seven antimicrobial resistance genes (>45% identity were predicted in LMB29 which includes a novel rifampicin ADP ribosyltransferase, arr-9, in plasmid pLMB157. The gene arr-9 was predicted on a genomic island with horizontal transferable potentials which may facilitate the rifampicin resistance dissemination. Future researches are needed to explore the pathogenesis of V. campbellii LMB29, but the availability of this genome sequence will certainly aid as a basis for further analysis.

  2. Phenotypic and Genetic Heterogeneity in Vibrio cholerae O139 Isolated from Cholera Cases in Delhi, India during 2001-2006

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    Raikamal Ghosh

    2016-08-01

    Full Text Available Incidence of epidemic Vibrio cholerae serogroup O139 has declined in cholera endemic countries. However, sporadic cholera caused by V. cholerae O139 with notable genetic changes is still reported from many regions. In the present study, 42 V. cholerae O139 strains isolated from 2001 to 2006 in Delhi, India, were retrospectively analyzed to understand their phenotype and molecular characteristics. The majority of isolates were resistant to ampicillin, furazolidone and nalidixic acid. Though the integrative conjugative element was detected in all the O139 isolates, the 2004-2006 isolates remained susceptible to co-trimoxazole, chloramphenicol, and streptomycin. Cholera toxin genotype 1 was present in the majority of the O139 isolates while few had type 3 or a novel type 4. In the cholera toxin encoding gene (ctx restriction fragment length polymorphism, the majority of the isolates harbored three copies of CTX element, of which one was truncated. In this study, the ctx was detected for the first time in the small chromosome of V. cholerae O139 and one isolate harbored 5 copies of CTX element, of which 3 were truncated. The ribotype BII pattern was found in most of the O139 isolates. Three V. cholerae O139 isolated in 2001 had a new ribotype BVIII. Pulsed-field gel electrophoresis analysis revealed clonal variation in 2001 isolates compared to the 2004-2006 isolates. Molecular changes in V. cholerae O139 have to be closely monitored as this information may help in understanding the changing genetic features of this pathogen in relation to the epidemiology of cholera.

  3. FIRST ISOLATION OF VIBRIO PROBIOTICUS FROM THE GUT OF PORTUNUS PELAGICUS (LINNAEUS, 1758 AND ITS PROBIOTIC COMPETENCY AGAINST SHELLFISH PATHOGENS

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    Mhd Ikhwanuddin

    2012-09-01

    Full Text Available In this study, the Vibrio probioticus was isolated from the gut of female Portunus pleagicus, identified through 16S rDNA gene sequencing, and was evaluated for its probiotic features. The probiont was tested in co-culture assay at various concentrations against three target shellfish pathogens including V. harveyi, V. parahaemolyticus and Pseudoalteromonas piscicida. In a co-culture assay it was apparent that the V. probioticus had shown to outcompete the growth of tested pathogenic bacteria due to its antibacteriocin properties. Different ratios produced different effect on tested pathogens, however, the highest ratio (pathogen:probiont was more effective in suppressing the pathogen. A maximum inhibition growth was recorded for V. harveyi and P. piscicida. The probionts could further be tested in vivo culture environment to understand the effectiveness in microbial control of P. pelagicus larviculture and other aquaculture species.

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

    Science.gov (United States)

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

    2009-11-01

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

  5. Isolation of Vibrio cholera El Tor Inaba From Lemna minor and Eichhornia crassipens Roots in Veracruz, Mexico.

    Science.gov (United States)

    Cordoba Aguilar, Edgar; Herrera Rivero, Marisol; Rubi, Alberto; Arroyo-Helguera, Omar; Coutino Rodriguez, Rocio

    2014-03-01

    During epidemic periods, the strain Vibrio cholera El Tor has been isolated from the aquatic macrophyte roots of Eichhornia crassipens and Lemna minor, suggesting that aquatic plants could be environmental reservoirs through either a non-specific association or a commensalism relationship. Therefore, it is important to understand V. cholera reservoirs in order to establish prevention strategies against this pathogen. Our interest was to determine whether V. cholera could be isolated and typified from L. minor and E. crassipens roots. From 2004 to 2005, plants were collected from various ecological niches and the roots were used to isolate V. cholera. Standard bacteriological, biochemical and serological tests were used for its typification. In five out of the nine ecological niches explored, we collected either L. minor or E. crassipens, as these specimens cohabited only in two niches. V. cholera was isolated from both L. minor and E. crassipens roots. The isolated V. cholera showed the same biochemical characteristics as the pure V. cholera strain which was used as a control. The isolated V. cholera corresponded to V. cholera O1 El Tor Inaba, which is the same serotype related to the last outbreak in Mexico. For first time V. cholera El Tor Inaba has been isolated several years after the last emergence of cholera in Mexico. A viable and cultivable V. cholera strain, sourced from freshwater niches in E. crassipens and L. minor roots, suggests the importance of these plants as a permanent aquatic reservoir for these organisms. The monitoring of E. crassipens and L. minor is the responsibility of health institutions in order to evaluate the ongoing risks.

  6. Phylodynamic analysis of clinical and environmental Vibrio cholerae isolates from Haiti reveals diversification driven by positive selection.

    Science.gov (United States)

    Azarian, Taj; Ali, Afsar; Johnson, Judith A; Mohr, David; Prosperi, Mattia; Veras, Nazle M; Jubair, Mohammed; Strickland, Samantha L; Rashid, Mohammad H; Alam, Meer T; Weppelmann, Thomas A; Katz, Lee S; Tarr, Cheryl L; Colwell, Rita R; Morris, J Glenn; Salemi, Marco

    2014-12-23

    Phylodynamic analysis of genome-wide single-nucleotide polymorphism (SNP) data is a powerful tool to investigate underlying evolutionary processes of bacterial epidemics. The method was applied to investigate a collection of 65 clinical and environmental isolates of Vibrio cholerae from Haiti collected between 2010 and 2012. Characterization of isolates recovered from environmental samples identified a total of four toxigenic V. cholerae O1 isolates, four non-O1/O139 isolates, and a novel nontoxigenic V. cholerae O1 isolate with the classical tcpA gene. Phylogenies of strains were inferred from genome-wide SNPs using coalescent-based demographic models within a Bayesian framework. A close phylogenetic relationship between clinical and environmental toxigenic V. cholerae O1 strains was observed. As cholera spread throughout Haiti between October 2010 and August 2012, the population size initially increased and then fluctuated over time. Selection analysis along internal branches of the phylogeny showed a steady accumulation of synonymous substitutions and a progressive increase of nonsynonymous substitutions over time, suggesting diversification likely was driven by positive selection. Short-term accumulation of nonsynonymous substitutions driven by selection may have significant implications for virulence, transmission dynamics, and even vaccine efficacy. Cholera, a dehydrating diarrheal disease caused by toxigenic strains of the bacterium Vibrio cholerae, emerged in 2010 in Haiti, a country where there were no available records on cholera over the past 100 years. While devastating in terms of morbidity and mortality, the outbreak provided a unique opportunity to study the evolutionary dynamics of V. cholerae and its environmental presence. The present study expands on previous work and provides an in-depth phylodynamic analysis inferred from genome-wide single nucleotide polymorphisms of clinical and environmental strains from dispersed geographic settings in

  7. Molecular Detection of the Three Major Pathogenic Vibrio Species from Seafood Products and Sediments in Tunisia Using Real-Time PCR.

    Science.gov (United States)

    Gdoura, Morsi; Sellami, Hanen; Nasfi, Hanen; Trabelsi, Rahma; Mansour, Sabeur; Attia, Touraya; Nsaibia, Siwar; Vallaeys, Tatiana; Gdoura, Radhouane; Siala, Mariam

    2016-12-01

    Vibrio spp. have emerged as a serious threat to human health worldwide. V. parahaemolyticus , V. cholerae , and V. vulnificus pose a considerable public health risk in Tunisia because they cause sporadic and epidemic foodborne infections associated with the consumption of raw or undercooked contaminated seafood. More recently, toxR-positive V. alginolyticus was also reported to be a potential source of contaminated seafood. A total of 247 samples, including 113 fishes ( Labrus viridis , Penaeus kerathurus , Diplodus annularis , Diplodus sparaillon , Scorparna porcus , Sarpa salpa , Dentex dentex , Scorparna scrofa , Sardinella aurita , Trachurus trachurus , Synodus saurus , Pagellus erythrinus , and Metapenaeus monoceros ), 83 clams ( Ruditapes decussatus species), 30 seawater samples, and 21 sediment samples were analyzed using traditional culture methods (ISO/TS 21872-1; International Organization for Standardization 2007) and a conventional PCR method for Vibrio spp. A rapid, sensitive, and highly reproducible real-time PCR assay was developed to detect the three major Vibrio spp. pathogenic for humans in Tunisian seafood products and sediments. A conventional culture method found 102 (41.3%) of 247 analyzed samples positive for Vibrio spp.; a conventional PCR method found 126 (51%) of the 247 samples positive. Real-time PCR assay found 126 (51.1%) samples positive; V. alginolyticus toxR was the most common, found in 99 (78.57%) of samples, followed by V. parahaemolyticus in 26 (20.63%) and V. cholerae in 1 (0.7%). All culture-positive samples were PCR positive. However, 24 samples that were positive by conventional PCR and real-time PCR were culture negative. Our findings indicate that retail seafood is commonly contaminated with Vibrio spp. and presents a potential risk to human health in Tunisia. These data also indicate that real-time PCR can provide sensitive species-specific detection of Vibrio spp. in seafood without prior isolation and characterization

  8. Clinical isolates of Vibrio cholerae O1 El Tor Ogawa of 2009 from Kolkata, India: preponderance of SXT element and presence of Haitian ctxB variant.

    Directory of Open Access Journals (Sweden)

    Braj M R N S Kutar

    Full Text Available BACKGROUND: Increase in the number of multidrug resistant pathogens and the accompanied rise in case fatality rates has hampered the treatment of many infectious diseases including cholera. Unraveling the mechanisms responsible for multidrug resistance in the clinical isolates of Vibrio cholerae would help in understanding evolution of these pathogenic bacteria and their epidemic potential. This study was carried out to identify genetic factors responsible for multiple drug resistance in clinical isolates of Vibrio cholerae O1, serotype Ogawa, biotype El Tor isolated from the patients admitted to the Infectious Diseases Hospital, Kolkata, India, in 2009. METHODOLOGY/PRINCIPAL FINDINGS: One hundred and nineteen clinical isolates of V. cholerae were analysed for their antibiotic resistance phenotypes. Antibiogram analysis revealed that majority of the isolates showed resistance to co-trimoxazole, nalidixic acid, polymixin B and streptomycin. In PCR, SXT integrase was detected in 117 isolates and its sequence showed 99% identity notably to ICEVchInd5 from Sevagram, India, ICEVchBan5 from Bangladesh and VC1786ICE sequence from Haiti outbreak among others. Antibiotic resistance traits corresponding to SXT element were transferred from the parent Vibrio isolate to the recipient E. coli XL-1 Blue cells during conjugation. Double-mismatch-amplification mutation assay (DMAMA revealed the presence of Haitian type ctxB allele of genotype 7 in 55 isolates and the classical ctxB allele of genotype 1 in 59 isolates. Analysis of topoisomerase sequences revealed the presence of mutation Ser83 → Ile in gyrA and Ser85→ Leu in parC. This clearly showed the circulation of SXT-containing V. cholerae as causative agent for cholera in Kolkata. CONCLUSIONS: There was predominance of SXT element in these clinical isolates from Kolkata region which also accounted for their antibiotic resistance phenotype typical of this element. DMAMA PCR showed them to be a mixture

  9. Molecular characterization of antibiotic resistant Vibrio harveyi isolated from shrimp aquaculture environment in the south east coast of India.

    Science.gov (United States)

    Stalin, Nattan; Srinivasan, Pappu

    2016-08-01

    Vibrio harveyi is a strategic human pathogen that occurs naturally in marine and estuarine environments. The pathogen is believed to cause acute septicemia, gastroenteritis, severe necrotizing soft-tissue infection, and high rate of lethality through ingestion of V. harveyi contaminated seafood. In this study, we isolated and characterized V. harveyi from water suspended sediment samples of black tiger shrimp ponds and from the sea coasts, in the east coast of the Bay of Bengal, India. Initial isolations of putative V. harveyi isolates were grown on thiosulfate-citrate-bill salts-sucrose agar (TCBS) plates for 36 h. Gram-negative and oxidase-positive colonies alone were selected and subsequently identified by 12 different conventional biochemical tests. The species specificity was confirmed by 16S rRNA, hemolysin and toxRvh genes were used through PCR targeted primers. Furthermore, genomic fingerprinting was carried out using randomly amplified polymorphic DNA fingerprinting, which showed that all the five V. harveyi were genetically distinct. From a total of 256 samples, a total of five strains of V. harveyi were isolated, of which three were from various shrimp ponds and two were from the coastal area. These five isolates were subjected to profiling against 15 antibiotics and the perusal results emphasized the V. harveyi resistance to ciprofloxacin, penicillin, rifampicin, and vancomycin compared to other tested antibiotics. The present findings were helpful in understanding the multiple antibiotics resistance of V. harveyi, which indicates the urgent need for targeted alternative biocontrol strategies to enhance the prospects of commercially viable shrimp cultivation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Antimicrobial effect of dietary oregano essential oil against Vibrio bacteria in shrimps

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    Gracia-Valenzuela M.H.

    2014-01-01

    Full Text Available The effect of dietary oregano essential oils on the growth of Vibrio bacteria in shrimps was evaluated. Shrimps were fed: (i food with oregano oil with a high level of thymol; (ii food with oregano oil with a high level of carvacrol, and (iii food without oregano oil (the control. The animals were infected by three species of Vibrio (vulnificus, parahaemolyticus and cholerae. The microbial counts of Vibrio species were significantly lower (p <0.05 in tissues from animals whose food was supplemented with oregano oil. We concluded that dietary supplementation of shrimps with oregano oil provides antimicrobial activity into the body of the penaeids.

  11. A genomic comparison of 13 symbiotic Vibrio fischeri isolates from the perspective of their host source and colonization behavior.

    Science.gov (United States)

    Bongrand, Clotilde; Koch, Eric J; Moriano-Gutierrez, Silvia; Cordero, Otto X; McFall-Ngai, Margaret; Polz, Martin F; Ruby, Edward G

    2016-12-01

    Newly hatched Euprymna scolopes squid obtain their specific light-organ symbionts from an array of Vibrio (Allivibrio) fischeri strains present in their environment. Two genetically distinct populations of this squid species have been identified, one in Kaneohe Bay (KB), and another in Maunaloa Bay (MB), Oahu. We asked whether symbionts isolated from squid in each of these populations outcompete isolates from the other population in mixed-infection experiments. No relationship was found between a strain's host source (KB or MB) and its ability to competitively colonize KB or MB juveniles in a mixed inoculum. Instead, two colonization behaviors were identified among the 11 KB and MB strains tested: a 'dominant' outcome, in which one strain outcompetes the other for colonization, and a 'sharing' outcome, in which two strains co-colonize the squid. A genome-level comparison of these and other V. fischeri strains suggested that the core genomic structure of this species is both syntenous and highly conserved over time and geographical distance. We also identified ~250 Kb of sequence, encoding 194 dispersed orfs, that was specific to those strains that expressed the dominant colonization behavior. Taken together, the results indicate a link between the genome content of V. fischeri strains and their colonization behavior when initiating a light-organ symbiosis.

  12. Historical report: first isolation of Vibrio Cholera serogroup 01 biovar El Tor serovar inaba during the cholerae epidemic in Peru - 1991

    OpenAIRE

    Bravo Cruz, Nora; Facultad de Ciencias Naturales y Matemática, Universidad Nacional Federico Villarreal. Bióloga. Exjefa del Departamento de Bacteriología, Centro Nacional de Salud Pública, Instituto Nacional de Salud.; Guillén, Alfredo; Facultad de Tecnología Médica, Universidad Nacional Federico Villarreal. Laboratorio de Microbiología, Clínica San Borja. Médico Microbiólogo.

    2011-01-01

    20 years ago, a new diarrheal disease was introduced in Peru and the Enteropathogens Reference Laboratory of the Instituto Nacional de Salud had an outstanding role in the isolation and rapid and timely identification of Vibrio cholerae. Cholera had not been seen before, but during the last week of January 1991 an outbreak of acute diarrhea was detected, presenting intense dehydration and some deaths. The epidemic affected, in the beginning, many locations of the peruvian coast. Some work...

  13. Isolation and characterization of a N4-like lytic bacteriophage infecting Vibrio splendidus, a pathogen of fish and bivalves.

    Directory of Open Access Journals (Sweden)

    Pantelis Katharios

    Full Text Available A novel virulent bacteriophage, vB_VspP_pVa5, infecting a strain of Vibrio splendidus was isolated from a sea-cage aquaculture farm in Greece, and characterized using microbiological methods and genomic analysis. Bacteriophage vB_VspP_pVa5 is a N4-like podovirus with an icosahedral head measuring 85 nm in length and a short non-contractile tail. The phage had a narrow host range infecting only the bacterial host, a latent period of 30 min and a burst size of 24 virions per infected bacterium. Its genome size was 78,145 bp and genomic analysis identified 107 densely-packed genes, 40 of which could be annotated. In addition to the very large virion encapsulated DNA-dependent RNA polymerase which is the signature of the N4-like genus, an interesting feature of the novel phage is the presence of a self-splicing group I intron in the thymidylate synthase gene. A tRNAStop interrupted by a ~2.5kb open reading frame-containing area was also identified. The absence of genes related to lysogeny along with the high efficacy observed during in vitro cell lysis trials, indicate that the vB_VspP_pVa5 is a potential candidate component in a bacteriophage cocktail suitable for the biological control of V. splendidus in aquaculture.

  14. Isolation and characterization of a N4-like lytic bacteriophage infecting Vibrio splendidus, a pathogen of fish and bivalves.

    Science.gov (United States)

    Katharios, Pantelis; Kalatzis, Panos G; Kokkari, Constantina; Sarropoulou, Elena; Middelboe, Mathias

    2017-01-01

    A novel virulent bacteriophage, vB_VspP_pVa5, infecting a strain of Vibrio splendidus was isolated from a sea-cage aquaculture farm in Greece, and characterized using microbiological methods and genomic analysis. Bacteriophage vB_VspP_pVa5 is a N4-like podovirus with an icosahedral head measuring 85 nm in length and a short non-contractile tail. The phage had a narrow host range infecting only the bacterial host, a latent period of 30 min and a burst size of 24 virions per infected bacterium. Its genome size was 78,145 bp and genomic analysis identified 107 densely-packed genes, 40 of which could be annotated. In addition to the very large virion encapsulated DNA-dependent RNA polymerase which is the signature of the N4-like genus, an interesting feature of the novel phage is the presence of a self-splicing group I intron in the thymidylate synthase gene. A tRNAStop interrupted by a ~2.5kb open reading frame-containing area was also identified. The absence of genes related to lysogeny along with the high efficacy observed during in vitro cell lysis trials, indicate that the vB_VspP_pVa5 is a potential candidate component in a bacteriophage cocktail suitable for the biological control of V. splendidus in aquaculture.

  15. Isolation of TDA-producing Phaeobacter strains from sea bass larval rearing units and their probiotic effect against pathogenic Vibrio spp. in Artemia cultures.

    Science.gov (United States)

    Grotkjær, Torben; Bentzon-Tilia, Mikkel; D'Alvise, Paul; Dourala, Nancy; Nielsen, Kristian Fog; Gram, Lone

    2016-05-01

    Fish-pathogenic Vibrio can cause large-scale crashes in marine larval rearing units and, since the use of antibiotics can result in bacterial antibiotic resistance, new strategies for disease prevention are needed. Roseobacter-clade bacteria from turbot larval rearing facilities can antagonize Vibrio anguillarum and reduce mortality in V. anguillarum-infected cod and turbot larvae. In this study, it was demonstrated that antagonistic Roseobacter-clade bacteria could be isolated from sea bass larval rearing units. In addition, it was shown that they not only antagonized V. anguillarum but also V. harveyi, which is the major bacterial pathogen in crustaceans and Mediterranean sea bass larvae cultures. Concomitantly, they significantly improved survival of V. harveyi-infected brine shrimp. 16S rRNA gene sequence homology identified the antagonists as Phaeobacter sp., and in silico DNA-DNA hybridization indicated that they could belong to a new species. The genomes contained genes involved in synthesis of the antibacterial compound tropodithietic acid (TDA), and its production was confirmed by UHPLC-TOFMS. The new Phaeobacter colonized live feed (Artemia) cultures and reduced Vibrio counts significantly, since they reached only 10(4)CFUmL(-1), as opposed to 10(8)CFUmL(-1) in non-Phaeobacter treated controls. Survival of V. anguillarum-challenged Artemia nauplii was enhanced by the presence of wild type Phaeobacter compared to challenged control cultures (89±1.0% vs 8±3.2%). In conclusion, TDA-producing Phaeobacter isolated from Mediterranean marine larviculture are promising probiotic bacteria against pathogenic Vibrio in crustacean live-feed cultures for marine fish larvae. Copyright © 2016 Elsevier GmbH. All rights reserved.

  16. Antiadhesive activity of poly-hydroxy butyrate biopolymer from a marine Brevibacterium casei MSI04 against shrimp pathogenic vibrios.

    Science.gov (United States)

    Kiran, George Seghal; Lipton, Anuj Nishanth; Priyadharshini, Sethu; Anitha, Kumar; Suárez, Lucia Elizabeth Cruz; Arasu, Mariadhas Valan; Choi, Ki Choon; Selvin, Joseph; Al-Dhabi, Naif Abdullah

    2014-08-13

    Vibrio pathogens are causative agents of mid-culture outbreaks, and early mortality syndrome and secondary aetiology of most dreadful viral outbreaks in shrimp aquaculture. Among the pathogenic vibrios group, Vibrio alginolyticus and V. harveyi are considered as the most significant ones in the grow-out ponds of giant black tiger shrimp Penaeus monodon in India. Use of antibiotics was banned in many countries due to the emergence of antibiotic-resistant strains and accumulation of residual antibiotics in harvested shrimp. There is an urgent need to consider the use of alternative antibiotics for the control of vibriosis in shrimp aquaculture. Biofilm formation is a pathogenic and/or establishment mechanism of Vibrio spp. This study aims to develop novel safe antibiofilm and/or antiadhesive process using PHB to contain vibrios outbreaks in shrimp aquaculture. In this study a poly-hydroxy butyrate (PHB) polymer producing bacterium Brevibacterium casei MSI04 was isolated from a marine sponge Dendrilla nigra and production of PHB was optimized under submerged-fermentation (SmF) conditions. The effect of carbon, nitrogen and mineral sources on PHB production and enhanced production of PHB by response surface methods were demonstrated. The maximum PHB accumulation obtained was 6.74 g/L in the optimized media containing 25 g/L starch as carbon source, 96 h of incubation, 35°C and 3% NaCl. The highest antiadhesive activity upto 96% was recorded against V. vulnificus, and V. fischeri, followed by 92% against V. parahaemolyticus and V. alginolyticus and 88% inhibition was recorded against V. harveyi. In this study, a thermostable biopolymer was chemically characterized as PHB based on 1HNMR spectra, FT-IR and GC-MS spectra. The NMR spectra revealed that the polymer was an isocratic homopolymer and it also confirmed that the compound was PHB. The antiadhesive activity of PHB was determined in microtitre plate assay and an effective concentration (EC) of PHB (200

  17. Genomic taxonomy of vibrios

    DEFF Research Database (Denmark)

    Thompson, Cristiane C.; Vicente, Ana Carolina P.; Souza, Rangel C.

    2009-01-01

    BACKGROUND: Vibrio taxonomy has been based on a polyphasic approach. In this study, we retrieve useful taxonomic information (i.e. data that can be used to distinguish different taxonomic levels, such as species and genera) from 32 genome sequences of different vibrio species. We use a variety...... analytical and bioinformatics tools will enable the most accurate species identification through genomic computational analysis. This endeavour will culminate in the birth of the online genomic taxonomy whereby researchers and end-users of taxonomy will be able to identify their isolates through a web...

  18. Prevalence and characterization of Vibrio cholerae isolated from shrimp products imported into Denmark

    DEFF Research Database (Denmark)

    Dalsgaard, A.; Bjergskov, T.; Jeppesen, V.F.

    1996-01-01

    , all originating from shrimp produced in aquaculture. Six isolated strains agglutinated in polyvalent O antisera, but did not agglutinate in Ogawa or Inaba antisera. The six strains were resistant to colistin and sulfisoxazole; three strains also showed resistance to ampicillin. None of the strains...

  19. High-frequency rugose exopolysaccharide production by Vibrio cholerae strains isolated in Haiti.

    Directory of Open Access Journals (Sweden)

    Mustafizur Rahman

    Full Text Available In October, 2010, epidemic cholera was reported for the first time in Haiti in over 100 years. Establishment of cholera endemicity in Haiti will be dependent in large part on the continued presence of toxigenic V. cholerae O1 in aquatic reservoirs. The rugose phenotype of V. cholerae, characterized by exopolysaccharide production that confers resistance to environmental stress, is a potential contributor to environmental persistence. Using a microbiologic medium promoting high-frequency conversion of smooth to rugose (S-R phenotype, 80 (46.5% of 172 V. cholerae strains isolated from clinical and environmental sources in Haiti were able to convert to a rugose phenotype. Toxigenic V. cholerae O1 strains isolated at the beginning of the epidemic (2010 were significantly less likely to shift to a rugose phenotype than clinical strains isolated in 2012/2013, or environmental strains. Frequency of rugose conversion was influenced by incubation temperature and time. Appearance of the biofilm produced by a Haitian clinical rugose strain (altered biotype El Tor HC16R differed from that of a typical El Tor rugose strain (N16961R by confocal microscopy. On whole-genome SNP analysis, there was no phylogenetic clustering of strains showing an ability to shift to a rugose phenotype. Our data confirm the ability of Haitian clinical (and environmental strains to shift to a protective rugose phenotype, and suggest that factors such as temperature influence the frequency of transition to this phenotype.

  20. Diversity of Vibrio navarrensis Revealed by Genomic Comparison: Veterinary Isolates Are Related to Strains Associated with Human Illness and Sewage Isolates While Seawater Strains Are More Distant.

    Science.gov (United States)

    Schwartz, Keike; Kukuc, Cindy; Bier, Nadja; Taureck, Karin; Hammerl, Jens A; Strauch, Eckhard

    2017-01-01

    Strains of Vibrio navarrensis are present in aquatic environments like seawater, rivers, and sewage. Recently, strains of this species were identified in human clinical specimens. In this study, V. navarrensis strains isolated from livestock in Germany were characterized that were found in aborted fetuses and/or placentas after miscarriages. The veterinary strains were analyzed using phenotypical and genotypical methods and compared to isolates from marine environments of the Baltic Sea and North Sea. The investigated phenotypical traits were similar in all German strains. Whole genome sequencing (WGS) was used to evaluate a phylogenetic relationship by performing a single nucleotide polymorphism (SNP) analysis. For the SNP analysis, WGS data of two American human pathogenic strains and two Spanish environmental isolates from sewage were included. A phylogenetic analysis of concatenated sequences of five protein-coding housekeeping genes (gyrB, pyrH, recA, atpA, and rpoB), was additionally performed. Both phylogenetic analyses reveal a greater distance of the environmental seawater strains to the other strains. The phylogenetic tree constructed from concatenated sequences of housekeeping genes places veterinary, human pathogenic and Spanish sewage strains into one cluster. Presence and absence of virulence-associated genes were investigated based on WGS data and confirmed by PCR. However, this analysis showed no clear pattern for the potentially pathogenic strains. The detection of V. navarrensis in human clinical specimens strongly suggests that this species should be regarded as a potential human pathogen. The identification of V. navarrensis strains in domestic animals implicates a zoonotic potential of this species. This could indicate a potential threat for humans, as according to the "One Health" concept, human, animal, and environmental health are linked. Future studies are necessary to search for reservoirs of these bacteria in the environment and/or in

  1. Production and Isolation of Amphibactin siderophores in Iron-stressed cultures of the marine bacteria Vibrio spp.

    Science.gov (United States)

    McLean, C.; Boiteau, R.; Bundy, R.; Gauglitz, J.; Repeta, D.

    2016-02-01

    Iron is an important micronutrient for marine microbes. Low concentrations of dissolved iron limit production in much of the ocean, putting pressure on microbial communities to develop efficient iron acquisition strategies. One such strategy is the production of siderophores, high affinity iron binding ligands, to facilitate iron uptake to meet their physiological iron quota. Recently, our lab has shown that amphibactins, siderophores with lipid side chains, are present in iron-deficient regions of the ocean. However, little is known about which organisms can utilize amphibactin bound iron. Here we describe a method to isolate amphibactins from laboratory cultures in order to identify the conditional stability constants and uptake rates of purified amphibactin compounds. We searched the National Center for Biotechnology Information database to identify microbial genomes containing homologous to the known amphibactin biosynthesis genes. Several of these strains were screened with high performance reverse-phase liquid chromatography electrospray ionization mass spectrometry (HPLC-ESIMS) to confirm amphibactin production. We then optimized amphibactin production for the strain Vibrio cyclitrophicus 1F53 under different shaking speeds and iron concentrations, using a chrome azurol S (CAS) assay to screen for siderophore abundance. Maximum production was found after 38 hours of shaking at 150-rpm, and with the addition of 10nM of desferrioxamine B to induce iron limitation. Amphibactins were extracted from the media by solid phase extraction and purified by reverse phase HPLC. The conditional stability constants for several amphibactins were then measured in seawater using competitive ligand exchange absorptive cathodic stripping voltammetry with salicylaldoxime as the added ligand. Future work will determine the uptake rates of these compounds by natural communities of marine bacteria, and give insight on the bioavailability of amphibactins in the marine environment.

  2. Vibrio and Pregnancy

    Science.gov (United States)

    ... 2013. Vibrio Infection. [Accessed January 2015]. Available at URL: http: / / www. cdc. gov/ vibrio/ index. html Centers for ... 2013. Vibrio parahaemolyticus. [Accessed January 2015]. Available at URL: http: / / www. cdc. gov/ vibrio/ vibriop. html Centers for ...

  3. Comparative genomic analysis of Vibrio parahaemolyticus: serotype conversion and virulence

    Directory of Open Access Journals (Sweden)

    Gil Ana I

    2011-06-01

    Full Text Available Abstract Background Vibrio parahaemolyticus is a common cause of foodborne disease. Beginning in 1996, a more virulent strain having serotype O3:K6 caused major outbreaks in India and other parts of the world, resulting in the emergence of a pandemic. Other serovariants of this strain emerged during its dissemination and together with the original O3:K6 were termed strains of the pandemic clone. Two genomes, one of this virulent strain and one pre-pandemic strain have been sequenced. We sequenced four additional genomes of V. parahaemolyticus in this study that were isolated from different geographical regions and time points. Comparative genomic analyses of six strains of V. parahaemolyticus isolated from Asia and Peru were performed in order to advance knowledge concerning the evolution of V. parahaemolyticus; specifically, the genetic changes contributing to serotype conversion and virulence. Two pre-pandemic strains and three pandemic strains, isolated from different geographical regions, were serotype O3:K6 and either toxin profiles (tdh+, trh- or (tdh-, trh+. The sixth pandemic strain sequenced in this study was serotype O4:K68. Results Genomic analyses revealed that the trh+ and tdh+ strains had different types of pathogenicity islands and mobile elements as well as major structural differences between the tdh pathogenicity islands of the pre-pandemic and pandemic strains. In addition, the results of single nucleotide polymorphism (SNP analysis showed that 94% of the SNPs between O3:K6 and O4:K68 pandemic isolates were within a 141 kb region surrounding the O- and K-antigen-encoding gene clusters. The "core" genes of V. parahaemolyticus were also compared to those of V. cholerae and V. vulnificus, in order to delineate differences between these three pathogenic species. Approximately one-half (49-59% of each species' core genes were conserved in all three species, and 14-24% of the core genes were species-specific and in different

  4. Vibrio sp. strain NM 10, isolated from the intestine of a Japanese coastal fish, has an inhibitory effect against Pasteurella piscicida.

    Science.gov (United States)

    Sugita, H; Matsuo, N; Hirose, Y; Iwato, M; Deguchi, Y

    1997-01-01

    Vibrio sp. strain NM 10 with an inhibitory activity against Pasteurella piscicida K-III was isolated from the intestine of a spotnape ponyfish (Leiognathus nuchalis). This bacterium efficiently produced an antibacterial substance after growth at 20 degrees C for 24 h on 1/5 PYBG agar prepared with 50% seawater at pHs of 7.5 to 9.0. The antibacterial substance was heat labile and proteinaceous, with a molecular mass of less than 5 kDa, possibly a bacteriocin or a bacteriocin-like substance. PMID:9406423

  5. Isolation and characterization of a N4-like lytic bacteriophage infecting Vibrio splendidus, a pathogen of fish and bivalves

    DEFF Research Database (Denmark)

    Katharios, Pantelis; Kalatzis, Panagiotis; Kokkari, Constantina

    2017-01-01

    A novel virulent bacteriophage, vB_VspP_pVa5, infecting a strain of Vibrio splendidus was isolated from a sea-cage aquaculture farm in Greece, and characterized using microbiological methods and genomic analysis. Bacteriophage vB_VspP_pVa5 is a N4-like podovirus with an icosahedral head measuring...... open reading frame–containing area was also identified. The absence of genes related to lysogeny along with the high efficacy observed during in vitro cell lysis trials, indicate that the vB_VspP_pVa5 is a potential candidate component in a bacteriophage cocktail suitable for the biological control...

  6. Vibrios associated with red tides caused by Mesodinium rubrum.

    OpenAIRE

    Romalde, J L; Barja, J L; Toranzo, A E

    1990-01-01

    Vibrios were isolated from red tides caused by Mesodinium rubrum and also throughout the year in the Ria de Pontevedra, Spain. The isolates were grouped into 14 phena by numerical toxonomy. Strains associated with red tides were restricted to four phena: phena I and II were Vibrio alginolyticus, and phena III and IV were Vibrio tubiashii and Vibrio anguillarum, respectively. V. anguillarum-like strains (phena V through XI) predominated throughout the year outside the red tide areas. Cytotoxic...

  7. Characterization and PCR Detection Of Binary, Pir-Like Toxins from Vibrio parahaemolyticus Isolates that Cause Acute Hepatopancreatic Necrosis Disease (AHPND in Shrimp.

    Directory of Open Access Journals (Sweden)

    Ratchanok Sirikharin

    Full Text Available Unique isolates of Vibrio parahaemolyticus (VPAHPND have previously been identified as the causative agent of acute hepatopancreatic necrosis disease (AHPND in shrimp. AHPND is characterized by massive sloughing of tubule epithelial cells of the hepatopancreas (HP, proposed to be induced by soluble toxins released from VPAHPND that colonize the shrimp stomach. Since these toxins (produced in broth culture have been reported to cause AHPND pathology in reverse gavage bioassays with shrimp, we used ammonium sulfate precipitation to prepare protein fractions from broth cultures of VPAHPND isolates for screening by reverse gavage assays. The dialyzed 60% ammonium sulfate fraction caused high mortality within 24-48 hours post-administration, and histological analysis of the moribund shrimp showed typical massive sloughing of hepatopancreatic tubule epithelial cells characteristic of AHPND. Analysis of the active fraction by SDS-PAGE revealed two major bands at marker levels of approximately 16 kDa (ToxA and 50 kDa (ToxB. Mass spectrometry analysis followed by MASCOT analysis revealed that both proteins had similarity to hypothetical proteins of V. parahaemolyticus M0605 (contig034 GenBank accession no. JALL01000066.1 and similarity to known binary insecticidal toxins called 'Photorhabdus insect related' proteins A and B (Pir-A and Pir-B, respectively, produced by the symbiotic, nematode bacterium Photorhabdus luminescens. In in vivo tests, it was shown that recombinant ToxA and ToxB were both required in a dose dependent manner to cause AHPND pathology, indicating further similarity to Pir-A and -B. A single-step PCR method was designed for detection of the ToxA gene and was validated using 104 bacterial isolates consisting of 51 VPAHPND isolates, 34 non-AHPND VP isolates and 19 other isolates of bacteria commonly found in shrimp ponds (including other species of Vibrio and Photobacterium. The results showed 100% specificity and sensitivity for

  8. The mosaic accessory gene structures of the SXT/R391-like integrative and conjugative elements derived from Vibrio spp. isolated from aquatic products and environment in the Yangtze River Estuary, China.

    Science.gov (United States)

    Song, Yuze; Yu, Pan; Li, Bailin; Pan, Yingjie; Zhang, Xiaojun; Cong, Jian; Zhao, Yinying; Wang, Hua; Chen, Lanming

    2013-09-30

    The emergence, resurgence and spread of human food-borne pathogenic Vibrios are one of the major contributors to disease burden and mortality particularly in developing countries with disputable sanitary conditions. Previous research on pathogenic Vibrio cholerae and Vibrio parahaemolitycus derived from clinical samples has proposed links between acquisition of virulence and multiple drug resistance traits and intercellular transmissibility of mobile genetic elements in the environment. To date, very few information is available on environmental Vibrio isolates. In this study, we characterized eleven Vibrio strains bearing the SXT/R391-like integrative and conjugative elements (ICEs) derived from aquatic products and environment in the Yangtze River Estuary, China. The eleven Vibrio strains were isolated in 2010 to 2011, and taxonomically identified, which included six Vibrio cholerae, three Vibrio parahaemolyticus, one Vibrio alginolyticus and one Vibrio natriegens. Most of the strains displayed strong resistance phenotypes to ampicillin, mercury and chromium. The majority of their ICEs, which belong to S and R exclusion system groups, contain ICEs-chromosome junction sequences and highly conserved core-genes required for ICE transfer. However, comparative sequence analysis uncovered interesting diversity in their mosaic accessory gene structures, which carry many novel genes that have not been described in any known ICEs to date. In addition, antibiotic resistance was transmitted by ICEVchChn6 and ICEVpaChn1 from V. cholerae, V. parahaemolyticus to E. coli MG1655 via conjugation, respectively. Our data also revealed that the ICEs characterized in this study are phylogenetically distant from most of the SXT/R391 ICEs reported previously, which may represent a novel cluster likely shaped by the ecological environment in the Yangtze River Estuary, China. This study constitutes the first investigation of ICEs-positive Vibrio spp. in the Yangze River Estuary, China

  9. The mosaic accessory gene structures of the SXT/R391-like integrative and conjugative elements derived from Vibrio spp. isolated from aquatic products and environment in the Yangtze River estuary, China

    Science.gov (United States)

    2013-01-01

    Background The emergence, resurgence and spread of human food-borne pathogenic Vibrios are one of the major contributors to disease burden and mortality particularly in developing countries with disputable sanitary conditions. Previous research on pathogenic Vibrio cholerae and Vibrio parahaemolitycus derived from clinical samples has proposed links between acquisition of virulence and multiple drug resistance traits and intercellular transmissibility of mobile genetic elements in the environment. To date, very few information is available on environmental Vibrio isolates. In this study, we characterized eleven Vibrio strains bearing the SXT/R391-like integrative and conjugative elements (ICEs) derived from aquatic products and environment in the Yangtze River Estuary, China. Results The eleven Vibrio strains were isolated in 2010 to 2011, and taxonomically identified, which included six Vibrio cholerae, three Vibrio parahaemolyticus, one Vibrio alginolyticus and one Vibrio natriegens. Most of the strains displayed strong resistance phenotypes to ampicillin, mercury and chromium. The majority of their ICEs, which belong to S and R exclusion system groups, contain ICEs-chromosome junction sequences and highly conserved core-genes required for ICE transfer. However, comparative sequence analysis uncovered interesting diversity in their mosaic accessory gene structures, which carry many novel genes that have not been described in any known ICEs to date. In addition, antibiotic resistance was transmitted by ICEVchChn6 and ICEVpaChn1 from V. cholerae, V. parahaemolyticus to E. coli MG1655 via conjugation, respectively. Our data also revealed that the ICEs characterized in this study are phylogenetically distant from most of the SXT/R391 ICEs reported previously, which may represent a novel cluster likely shaped by the ecological environment in the Yangtze River Estuary, China. Conclusions This study constitutes the first investigation of ICEs-positive Vibrio spp. in the

  10. Presence of T3SS2β genes in trh⁺ Vibrio parahaemolyticus isolated from seafood harvested along Mangalore coast, India.

    Science.gov (United States)

    Kumar, B K; Deekshit, V K; Rai, P; Shekar, M; Karunasagar, I; Karunasagar, I

    2014-05-01

    Vibrio parahaemolyticus is a seafood-borne pathogen autochthonous to the marine and estuarine ecosystem, responsible for gastroenteritis when contaminated raw seafood is consumed. The pathogenicity has been associated with thermostable direct haemolysin (TDH) and TDH-related haemolysin (TRH). Of late, the presence of T3SS2α and T3SS2β gene clusters has been well documented in clinical isolates of Vibrio parahaemolyticus and known to play an essential role in pathogenesis. However, reports on the presence of T3SSβ genes in V. parahaemolyticus isolated from the seafood and/or environmental samples are scanty. In this study, we have identified and analysed the distribution of the T3SS2β genes in V. parahaemolyticus isolated from seafood harvested along southwest coast of India. Results showed that T3SS2β genes are solely associated with trh⁺ and tdh⁺ /trh⁺ strains of V. parahaemolyticus. Reverse transcriptase PCR (RT-PCR) showed that the T3SS2β genes identified in trh⁺ V. parahaemolyticus were transcriptionally active. To our knowledge, this study appears to be the first description on the presence of T3SS2β-positive V. parahaemolyticus isolated from seafood in India. The study of T3SS2 along with other virulence factors will help in better understanding of the risk of seafood-borne illness due to V. parahaemolyticus. T3SSs (α or β) are the important virulence factors of Vibrio parahaemolyticus that contribute to their pathogenicity in humans. This study demonstrated the presence of T3SS2β genes in V. parahaemolyticus isolated from the seafood harvested along Mangalore coast. RT-PCR showed that the T3SS2β genes identified in seafood isolates of V. parahaemolyticus were found to be functional. To the best of our knowledge, this is the first description of T3SS2β genes in trh⁺ V. parahaemolyticus isolated from seafood in India. The presence of T3SS2 along with other virulence factors such as TDH and/or TRH highlights a potential health risk for

  11. Vibrio spp. ISOLATED FROM SHRIMPS AND WATER FROM A MARINE FARM IN PERNAMBUCO, BRAZIL Víbrios en el agua y en las gambas del mar (Litopenaeus vannamei, Boone, 1931 cultivado en Pernambuco Vibrio spp. ISOLADOS DE CAMARÃO E ÁGUA DE CULTIVO DE FAZENDA MARINHA EM PERNAMBUCO

    Directory of Open Access Journals (Sweden)

    Carlos André Bezerra Alves

    2009-12-01

    Full Text Available

    Water and shrimp samples were collected monthly, during all cultivation phases, in three located farms at Pernambuco coast, on winter and summer, for Vibrio spp. quantification and identification. The counting’s were correlated, through mathematical models (P<0.05, with the variables season, water biochemical parameters, wet mount, histopathology exam, toxins presence and handling techniques used. Just the variable cultivation time interfered at total counting of Vibrio spp. in all samples were obtained countings that varied 0.1x 10 to 6.2 x 103 UFC/mL in water, of 7.0 x 10 to 8.2 x 105 UFC/g in powder-larva, of 1.1 x 10 to 1.1 x 105 UFC/mL in hemolymph and of 2.5 x 102 to 1.1 x 106UFC/g in hepatopancreas. The species V. mediterranei (1%, V. mimicus (1.25%, V. fischeri (4.25%, V. cincinnatiensis (4.25%, V. metschnikovii (4.25%, V. proteolyticus (5.5%, V. harveyi (5.5%, V. hollisae (5.5%, V. carchariae (7%, V. vulnificus (8.5%, V. damsela (8.5%, V. parahaemolyticus (13%, V. fluvialis (15%, V. anguillarum (16.5% were identified. It is concluded that the Vibrionaceae load increases proportionally with cultivation time, due to the organic matter increment, what can turn the susceptible animals to the infection for vibrios.

    KEY WORD: Vibrionaceae, Vibrio spp., shrimp and pond water.
    El agua y muestras de la gamba eran la publicación mensual reunido, durante todas las fases del cultivo, en tres granjas localizadas a la costa de Pernambuco, en invierno y verano, para la cuantificación e identificación el de Vibrio spp.. El contando fueron puestos en correlación, a través de los modelos matemáticos (P <0,05, con la estación de las variables, agua que los parámetros bioquímicos, montaña mojada, examen del histopatologia, presencia de las toxinas y técnicas manejando usaron. Simplemente el tiempo del cultivo inconstante interfirió a total que

  12. SURVEY ON V. CHOLERAE, V. VULNIFICUS AND V. PARAHAEMOLYTICUS IN BIVALVE MOLLUSCS OF THE ADRIATIC SEA AND PROPOSAL OF AN ANALYTICAL PROTOCOL

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    M.L. Valeri

    2009-06-01

    Full Text Available Bivalve molluscs from Adriatic sea were analyzed for V. parahaemolyticus, V. cholerae e V. vulnificus presence. The isolates on TCBS Agar and m-CPC Agar were selected on the basis of a new biochemical screening, that showed a good performance, because among 2344 strains from primary culture only 237 (10% were presumptively assigned to the species of interest. The PCR analyses was performed for the target genes toxR hlyA, ctxA, tcpI (V. cholerae, toxR, tl, tdh, trh (V. parahaemolyticus, vvhA and viuB (V. vulnificus. Among the 9 strains confirmed to belong to V. parahaemolyticus specie, 6 were sucrose positive. On 215 samples of molluscs only 5 resulted positive for V. parahaemolyticus being toxR+, tl+, although non pathogenic (tdh-, trh-, and none for V. cholerae e V. vulnificus.

  13. Genomic taxonomy of vibrios

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    Iida Tetsuya

    2009-10-01

    Full Text Available Abstract Background Vibrio taxonomy has been based on a polyphasic approach. In this study, we retrieve useful taxonomic information (i.e. data that can be used to distinguish different taxonomic levels, such as species and genera from 32 genome sequences of different vibrio species. We use a variety of tools to explore the taxonomic relationship between the sequenced genomes, including Multilocus Sequence Analysis (MLSA, supertrees, Average Amino Acid Identity (AAI, genomic signatures, and Genome BLAST atlases. Our aim is to analyse the usefulness of these tools for species identification in vibrios. Results We have generated four new genome sequences of three Vibrio species, i.e., V. alginolyticus 40B, V. harveyi-like 1DA3, and V. mimicus strains VM573 and VM603, and present a broad analyses of these genomes along with other sequenced Vibrio species. The genome atlas and pangenome plots provide a tantalizing image of the genomic differences that occur between closely related sister species, e.g. V. cholerae and V. mimicus. The vibrio pangenome contains around 26504 genes. The V. cholerae core genome and pangenome consist of 1520 and 6923 genes, respectively. Pangenomes might allow different strains of V. cholerae to occupy different niches. MLSA and supertree analyses resulted in a similar phylogenetic picture, with a clear distinction of four groups (Vibrio core group, V. cholerae-V. mimicus, Aliivibrio spp., and Photobacterium spp.. A Vibrio species is defined as a group of strains that share > 95% DNA identity in MLSA and supertree analysis, > 96% AAI, ≤ 10 genome signature dissimilarity, and > 61% proteome identity. Strains of the same species and species of the same genus will form monophyletic groups on the basis of MLSA and supertree. Conclusion The combination of different analytical and bioinformatics tools will enable the most accurate species identification through genomic computational analysis. This endeavour will culminate in

  14. Rapid identification of Vibrio parahaemolyticus isolated from shellfish, sea water and sediments of the Khnifiss lagoon, Morocco, by MALDI-TOF mass spectrometry.

    Science.gov (United States)

    Malainine, S M; Moussaoui, W; Prévost, G; Scheftel, J-M; Mimouni, R

    2013-05-01

    We establish the presence of Vibrio parahaemolyticus and deepen the comparison of isolates using MALDI-TOF MS for the typing of isolates originating from the Khnifiss lagoon (Morocco). Amongst 48 samples from sea water, sediment and shellfish isolated from different sites of Khnifiss lagoon, Morocco, we obtained 22 isolates of V. parahaemolyticus identified by Vitek 2™ System (bioMérieux) and MALDI Biotyper™ (Bruker Daltonics). All isolates were highly resistant to ampicillin and ticarcillin, moderately resistant to cefalotin, but sensitive to 16 other antimicrobials tested. MALDI-TOF MS was used to discriminate between closely related environmental strains of V. parahaemolyticus. A clustering and distribution based on MALDI-TOF spectra were generated using the BioTyper 1.1™ software. Despite low diversity in regard to the biochemical characteristics and antimicrobial resistance, the isolates evoke a larger biodiversity when analysed through mass spectra of abundant proteins. Different evaluations of a cut-off value showed that, when placed at a 10% threshold of the whole diversity, isolates differed by at least three mass peaks. © 2013 The Society for Applied Microbiology.

  15. Diversity of Clinical and Environmental Isolates of Vibrio cholerae in Natural Transformation and Contact-Dependent Bacterial Killing Indicative of Type VI Secretion System Activity.

    Science.gov (United States)

    Bernardy, Eryn E; Turnsek, Maryann A; Wilson, Sarah K; Tarr, Cheryl L; Hammer, Brian K

    2016-05-01

    The bacterial pathogen Vibrio cholerae can occupy both the human gut and aquatic reservoirs, where it may colonize chitinous surfaces that induce the expression of factors for three phenotypes: chitin utilization, DNA uptake by natural transformation, and contact-dependent bacterial killing via a type VI secretion system (T6SS). In this study, we surveyed a diverse set of 53 isolates from different geographic locales collected over the past century from human clinical and environmental specimens for each phenotype outlined above. The set included pandemic isolates of serogroup O1, as well as several serogroup O139 and non-O1/non-O139 strains. We found that while chitin utilization was common, only 22.6% of the isolates tested were proficient at chitin-induced natural transformation, suggesting that transformation is expendable. Constitutive contact-dependent killing of Escherichia coli prey, which is indicative of a functional T6SS, was rare among clinical isolates (only 4 of 29) but common among environmental isolates (22 of 24). These results bolster the pathoadaptive model in which tight regulation of T6SS-mediated bacterial killing is beneficial in a human host, whereas constitutive killing by environmental isolates may give a competitive advantage in natural settings. Future sequence analysis of this set of diverse isolates may identify previously unknown regulators and structural components for both natural transformation and T6SS. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  16. Caracterización de aislamientos de Vibrio cholerae no-O1, no-O139 asociados a cuadros de diarrea Characterization of Vibrio cholerae non-O1 and non-O139 isolates associated with diarrhea

    Directory of Open Access Journals (Sweden)

    S. González Fraga

    2009-03-01

    Full Text Available La infección por Vibrio cholerae, el agente causal del cólera, se trasmite al hombre por ingestión de agua y alimentos contaminados. Aunque son los serogrupos O1 y O139 los que habitualmente se asocian al cólera epidémico, los aislamientos de otros serogrupos también son causales de gastroenteritis e infecciones extra-intestinales. Durante el período 2003-2005, se investigó la presencia de V. cholerae en la materia fecal de niños con diarrea atendidos en el Hospital del Niño Jesús, Tucumán. Se recuperaron 34 aislamientos de V. cholerae no-O1, no-O139. Se determinaron sus perfiles de virulencia por PCR, la sensibilidad a los antimicrobianos y la diversidad genética por electroforesis en campo pulsado. Se obtuvieron ocho perfiles de virulencia, aunque ningún aislamiento fue positivo para la toxina colérica ni para la toxina termoestable. Cuatro aislamientos fueron positivos para el sistema de secreción de tipo tres. El 17,6% de los aislamientos fueron resistentes o de sensibilidad intermedia a ampicilina y el 5,9% fueron resistentes a trimetoprima-sulfametoxazol. Los aislamientos resultaron muy diversos: se hallaron 27 patrones distintos en 29 aislamientos tipificables por electroforesis en campo pulsado. A pesar de su baja incidencia, V. cholerae continúa siendo un agente causal de diarrea en niños, los que se ven afectados por una amplia variedad de cepas circulantes.Vibrio cholerae, etiologic agent of cholera, is transmitted to humans by ingestion of contaminated food or water. Even though serogroups O1 and O139 are the ones usually associated to epidemic cholera, isolates from other serogroups also cause gastroenteritis and extraintestinal infections. During the period 2003-2005, presence of V. cholerae in stools was investigated in children with diarrhea that seaked assistance at the Niño Jesús Hospital in Tucumán. Thirty four isolates of V. cholerae non-O1, non-O139 were recovered. We characterized the isolates studying

  17. Occurrences of pathogenic Vibrio parahaemolyticus from Vellar ...

    African Journals Online (AJOL)

    Vibrio parahaemolyticus is the predominant seafood pathogen associated with human gastroenteritis. Samples were collected from Vellar estuary, shrimp ponds and shrimp for characterization of V. parahaemolyticus. A total of 26 blue green centre (BG) Vibrio strains were isolated and characterized through biochemical ...

  18. Virulence-associated factors in Vibrio cholerae non-O1/non-O139 and V. mimicus strains isolated in ornamental fish species.

    Science.gov (United States)

    Zago, V; Zambon, M; Civettini, M; Zaltum, O; Manfrin, A

    2017-12-01

    During recent decades, ornamental fish have proven to be one of the fastest growing categories of pets in Europe. In this framework, we evaluated both the potential pathogenic and zoonotic risks caused by 53 Vibrio cholerae non-O1/non-O139 and a Vibrio mimicus strain isolated from ornamental fish species mostly originating from South-East Asia countries between 2000 and 2015 in Italy. All the strains were firstly identified at species level by biochemical, phylogenetic and mass spectrometry (matrix-assisted laser desorption ionization time of flight) methods, and then studied to reveal the presence of the main virulence and colonization-associated factors, as ctxA, ace, zot, stn/sto, toxR, rtxA, hlyA and tcpA by multiplex and single endpoint PCR assays. Findings showed that 21 of 54 strains harboured at least one virulence factor with a predominance for the toxR+ , rtxA+ and hlyAET+ genotype. Interestingly, the V. mimicus strain harboured the colonization factor and the CTX prophage receptor, tcpA, indicating the ability to capture and integrate it in its genome increasing its pathogenicity. Although these enterotoxins can sporadically cause gastroenteritis, the results highlight their probable involvement in causing severe implications for public health, suggesting the need for an European microbiological monitoring. © 2017 John Wiley & Sons Ltd.

  19. Non-Cholera Vibrios: The Microbial Barometer of Climate Change.

    Science.gov (United States)

    Baker-Austin, Craig; Trinanes, Joaquin; Gonzalez-Escalona, Narjol; Martinez-Urtaza, Jaime

    2017-01-01

    There is a growing interest in the role of climate change in driving the spread of waterborne infectious diseases, such as those caused by bacterial pathogens. One particular group of pathogenic bacteria - vibrios - are a globally important cause of diseases in humans and aquatic animals. These Gram-negative bacteria, including the species Vibrio vulnificus, Vibrio parahaemolyticus and Vibrio cholerae, grow in warm, low-salinity waters, and their abundance in the natural environment mirrors ambient environmental temperatures. In a rapidly warming marine environment, there are greater numbers of human infections, and most notably outbreaks linked to extreme weather events such as heatwaves in temperate regions such as Northern Europe. Because the growth of pathogenic vibrios in the natural environment is largely dictated by temperature, we argue that this group of pathogens represents an important and tangible barometer of climate change in marine systems. We provide a number of specific examples of the impacts of climate change on this group of bacteria and their associated diseases, and discuss advanced strategies to improve our understanding of these emerging waterborne diseases through the integration of microbiological, genomic, epidemiological, climatic, and ocean sciences. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

  20. Marine Bacteria with antimicrobials capacity isolated from cultures of bivalve mollusks

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    Fabiola Pellon

    2014-06-01

    Full Text Available Microorganisms have commonly been studied as producers of antibacterial substances; yet they are also considered producers of antifungic, antiviral, antiparasitic, citotoxics and inhibitory of other forms of cellular growth substances. This paper describes the isolation, inhibitory potential and phenotipic characterization of native bacterial strains associated to bivalve mollusks such as Argopecten purpuratus “concha de abanico” and Crassostrea gigas “ostra” in cultivation systems. From 345 marine strains collected, 20 strains were recovered that had the ability of inhibiting a wide spectrum of fish, mollusks and shellfish pathogenic bacteria; being the most sensitive pathogens Aeromonas sobria P-281, Aeromonas hydrophila ATCC 7966, Vibrio vulnificus ATCC 27562 and Vibrio parahaemolyticus ATCC 17803. The phenotipic characterization of this strains with inhibitory capacity allowed the identification of the following genera: Vibrio (40%, Aeromonas (15%, Flavobacterium (10%, Pseudomonas (5%, Moraxella (5%, Flexibacter (5%. A 20% could not be identified. The results suggest that the isolated bacteria could be used as probiotics agents for the biological control of pathogens from marine organisms of interest in mariculture.

  1. The Lake Chad Basin, an Isolated and Persistent Reservoir of Vibrio cholerae O1: A Genomic Insight into the Outbreak in Cameroon, 2010.

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    Rolf S Kaas

    Full Text Available The prevalence of reported cholera was relatively low around the Lake Chad basin until 1991. Since then, cholera outbreaks have been reported every couple of years. The objective of this study was to investigate the 2010/2011 Vibrio cholerae outbreak in Cameroon to gain insight into the genomic make-up of the V. cholerae strains responsible for the outbreak. Twenty-four strains were isolated and whole genome sequenced. Known virulence genes, resistance genes and integrating conjugative element (ICE elements were identified and annotated. A global phylogeny (378 genomes was inferred using a single nucleotide polymorphism (SNP analysis. The Cameroon outbreak was found to be clonal and clustered distant from the other African strains. In addition, a subset of the strains contained a deletion that was found in the ICE element causing less resistance. These results suggest that V. cholerae is endemic in the Lake Chad basin and different from other African strains.

  2. The Lake Chad Basin, an Isolated and Persistent Reservoir of Vibrio cholerae O1: A Genomic Insight into the Outbreak in Cameroon, 2010

    DEFF Research Database (Denmark)

    Kaas, Rolf Sommer; Ngandjio, Antoinette; Nzouankeu, Ariane

    2016-01-01

    a single nucleotide polymorphism (SNP) analysis. The Cameroon outbreak was found to be clonal and clustered distant from the other African strains. In addition, a subset of the strains contained a deletion that was found in the ICE element causing less resistance. These results suggest that V. cholerae......The prevalence of reported cholera was relatively low around the Lake Chad basin until 1991. Since then, cholera outbreaks have been reported every couple of years. The objective of this study was to investigate the 2010/2011 Vibrio cholerae outbreak in Cameroon to gain insight into the genomic...... make-up of the V. cholerae strains responsible for the outbreak. Twenty-four strains were isolated and whole genome sequenced. Known virulence genes, resistance genes and integrating conjugative element (ICE) elements were identified and annotated. A global phylogeny (378 genomes) was inferred using...

  3. Factors associated with virulence and survival in environmental and clinical isolates of Vibrio cholerae O1 and non O1 in Romania.

    Science.gov (United States)

    Israil, Anca; Balotescu, Carmen; Bucurenci, Nadia; Năcescu, Nadia; Cedru, Claudia; Popa, Cornelia; Ciufecu, C

    2003-01-01

    Four hundred ninety seven strains of Vibrio cholerae selected from isolates in Romania in the last decade 1990-1999 were investigated for antibiotic resistance and for classical and putative virulence factors. V. cholerae O1 strains predominated in clinical cases and non O1 strains in the environment, excepting in 1992 when non O1 strains were frequent in clinical and environmental sources. V. cholerae O1 strains previously susceptible to tetracycline acquired clinically significant resistance to this drug during 1993-1994, but this trend was reversed in 1995, following the introduction of nalidixic acid in cholera treatment in 1994. V. cholerae O1 and non O1 clinical isolates acquired simultaneous resistance to the vibriostatic agent O/129 and cotrimoxazole during 1994-1995. High levels of intrinsic resistance to multiple antibiotics were exhibited by all strains examined. The presence of cholera toxin (CT) was concentrated in clinical V. cholerae O1 strains and was substituted in clinical non O1 strains by four putative virulence markers (Kanagawa haemolysin, slime, lipase, and colonial opacity). Colonial opacity (30%) was present only in clinical isolates of V. cholerae non O1. Pigmentogenesis (11.7%) has present only in environmental sources. Antibioresistance profiles differ for V. cholerae O1 and non O1 strains with respect to their source of isolation. This aspect may imply a role in virulence and survival of V. cholerae in the natural environment where they may serve as a reservoir of virulence and multiple drug resistance genes.

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

    Science.gov (United States)

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

    2010-11-01

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

  5. Trh (tdh-/trh+) gene analysis of clinical, environmental and food isolates of Vibrio parahaemolyticus as a tool for investigating pathogenicity.

    Science.gov (United States)

    Leoni, Francesca; Talevi, Giulia; Masini, Laura; Ottaviani, Donatella; Rocchegiani, Elena

    2016-05-16

    Sequencing analysis of the trh gene encoding the TDH-related haemolysin of tdh-/trh+ Vibrio parahaemolyticus isolated in Italy between 2002 and 2011 from clinical, environmental, and food samples revealed the presence of the trh2 variant in all isolates. The trh2 of the clinical isolate was 100% identical to other clinical tdh-/trh2 V. parahaemolyticus from Europe. Nucleotide and amino acid differences in the trh2 sequences of clinical isolates from Italy and other countries allowed a differentiation of the clinical strains from the majority of environmental or food strains isolated in Italy. Aspartic acid and isoleucine at positions 113 and 115, encoded by nucleotide triplets GAT and ATT at positions 337-339 and 343-345 of the complete trh gene sequence, were present in clinical strains from Europe (Italy, Norway and Germany), Asia and the United States. Only 35.5% of the tdh-/trh2 V. parahaemolyticus of environmental or food origin from Italy shared the same triplets/amino acid detected in clinical isolates, while 64.5% of isolates from the marine environment were different from those of clinical origins, demonstrating that differences occur amongst the trh2 sequences of strains from the environment and these polymorphisms may differentiate potentially pathogenic from less or non-pathogenic cultures found in the environment and seafood. In addition the distribution of T3SS2 genes was investigated in this group of tdh-/trh+ V. parahaemolyticus from different sources and in three clinical tdh+/trh- V. parahaemolyticus isolates. All tdh-/trh+ V. parahaemolyticus of environmental or food source, independent of year of isolation or geographical origin, amplified all the screened T3SS2β genes and tested negative to PCR assays for all five T3SS2α genes, as the tdh-/trh+ clinical V. parahaemolyticus isolate. The vopC genes, encoding for one of the effector proteins of T3SS2, were partially sequenced and compared to clinical tdh-/trh+ and tdh+/trh+ V. parahaemolyticus

  6. Comparative genome analysis of VSP-II and SNPs reveals heterogenic variation in contemporary strains of Vibrio cholerae O1 isolated from cholera patients in Kolkata, India.

    Science.gov (United States)

    Imamura, Daisuke; Morita, Masatomo; Sekizuka, Tsuyoshi; Mizuno, Tamaki; Takemura, Taichiro; Yamashiro, Tetsu; Chowdhury, Goutam; Pazhani, Gururaja P; Mukhopadhyay, Asish K; Ramamurthy, Thandavarayan; Miyoshi, Shin-Ichi; Kuroda, Makoto; Shinoda, Sumio; Ohnishi, Makoto

    2017-02-01

    Cholera is an acute diarrheal disease and a major public health problem in many developing countries in Asia, Africa, and Latin America. Since the Bay of Bengal is considered the epicenter for the seventh cholera pandemic, it is important to understand the genetic dynamism of Vibrio cholerae from Kolkata, as a representative of the Bengal region. We analyzed whole genome sequence data of V. cholerae O1 isolated from cholera patients in Kolkata, India, from 2007 to 2014 and identified the heterogeneous genomic region in these strains. In addition, we carried out a phylogenetic analysis based on the whole genome single nucleotide polymorphisms to determine the genetic lineage of strains in Kolkata. This analysis revealed the heterogeneity of the Vibrio seventh pandemic island (VSP)-II in Kolkata strains. The ctxB genotype was also heterogeneous and was highly related to VSP-II types. In addition, phylogenetic analysis revealed the shifts in predominant strains in Kolkata. Two distinct lineages, 1 and 2, were found between 2007 and 2010. However, the proportion changed markedly in 2010 and lineage 2 strains were predominant thereafter. Lineage 2 can be divided into four sublineages, I, II, III and IV. The results of this study indicate that lineages 1 and 2-I were concurrently prevalent between 2007 and 2009, and lineage 2-III observed in 2010, followed by the predominance of lineage 2-IV in 2011 and continued until 2014. Our findings demonstrate that the epidemic of cholera in Kolkata was caused by several distinct strains that have been constantly changing within the genetic lineages of V. cholerae O1 in recent years.

  7. Characterization of VCC-1, a Novel Ambler Class A Carbapenemase from Vibrio cholerae Isolated from Imported Retail Shrimp Sold in Canada.

    Science.gov (United States)

    Mangat, Chand S; Boyd, David; Janecko, Nicol; Martz, Sarah-Lynn; Desruisseau, Andrea; Carpenter, Michael; Reid-Smith, Richard J; Mulvey, Michael R

    2016-01-11

    One of the core goals of the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) is to monitor major meat commodities for antimicrobial resistance. Targeted studies with methodologies based on core surveillance protocols are used to examine other foods, e.g., seafood, for antimicrobial resistance to detect resistances of concern to public health. Here we report the discovery of a novel Ambler class A carbapenemase that was identified in a nontoxigenic strain of Vibrio cholerae (N14-02106) isolated from shrimp that was sold for human consumption in Canada. V. cholerae N14-02106 was resistant to penicillins, carbapenems, and monobactam antibiotics; however, PCR did not detect common β-lactamases. Bioinformatic analysis of the whole-genome sequence of V. cholerae N14-02106 revealed on the large chromosome a novel carbapenemase (referred to here as VCC-1, for Vibrio cholerae carbapenemase 1) with sequence similarity to class A enzymes. Two copies of blaVCC-1 separated and flanked by ISVch9 (i.e., 3 copies of ISVch9) were found in an acquired 8.5-kb region inserted into a VrgG family protein gene. Cloned blaVCC-1 conferred a β-lactam resistance profile similar to that in V. cholerae N14-02106 when it was transformed into a susceptible laboratory strain of Escherichia coli. Purified VCC-1 was found to hydrolyze penicillins, 1st-generation cephalosporins, aztreonam, and carbapenems, whereas 2nd- and 3rd-generation cephalosporins were poor substrates. Using nitrocefin as a reporter substrate, VCC-1 was moderately inhibited by clavulanic acid and tazobactam but not EDTA. In this report, we present the discovery of a novel class A carbapenemase from the food supply. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  8. Power plays: iron transport and energy transduction in pathogenic vibrios.

    Science.gov (United States)

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

    2011-06-01

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

  9. Water sources as reservoirs of Vibrio cholerae O1 and non-O1 strains in Bepanda, Douala (Cameroon): relationship between isolation and physico-chemical factors.

    Science.gov (United States)

    Akoachere, Jane-Francis Tatah Kihla; Mbuntcha, Christelle Kwedjeu Pulcherie

    2014-07-30

    Cholera has been endemic in Douala since 1971. Most outbreaks start from Bepanda, an overcrowded neighbourhood with poor hygiene and sanitary conditions. We investigated water sources in Bepanda as reservoirs of Vibrio cholerae, the causative agent of cholera, determined its antibiotic susceptibility and some physico-chemical characteristics that could maintain the endemicity of this organism in Bepanda. Three hundred and eighteen water samples collected from 45 wells, 8 taps and 1 stream from February to July 2009 were analyzed for V. cholerae using standard methods. Isolates were characterized morphologically, biochemically and serologically. The disc diffusion technique was employed to investigate antibiotic susceptibility. Differences in prevalence of organism between seasons were analysed. Correlation strength and direction of association between physico-chemical parameters and occurrence of V. cholerae was analyzed using the Kendall tau_b non-parametric correlation. This was further confirmed with the forward-stepwise binary logistic regression. Eighty-seven (27.4%) samples were positive for V. cholerae. Isolation was highest from wells. The organism was isolated in the rainy season and dry season but the frequency of isolation was significantly higher (χ2 = 7.009, df = 1, P = 0.008) in the rainy season. Of the 96 confirmed V. cholerae isolates, 32 (33.3%) belonged to serogroup O1 and 64 (66.6%) were serogroup non-O1/non-O139. Isolates from tap (municipal water) were non-O1/non-O139 strains. Salinity had a significant positive correlation with isolation in the dry season (+0.267, P = 0.015) and rainy season (+0.223, P = 0.028). The forward-stepwise method of binary logistic regression indicated that as pH (Wald = 11.753, df = 1), P = 0.001) increased, odds of isolation of V. cholerae also increased (B = 1.297, S.E = 0.378, Exp(B) = 3.657). All isolates were sensitive to ciprofloxacin and ofloxacin. Multi-drug resistance was predominant among the non-O1/non

  10. Desarrollo de métodos moleculares para la detección y caracterización de bacterias patógenas emergentes del género Vibrio en aguas y alimentos

    OpenAIRE

    Cañigral Cárcel, Irene

    2011-01-01

    Vibrio parahaemolyticus y Vibrio vulnificus son microorganismos patógenos pertenecientes a la familia Vibrionaceae y al género Vibrio. Son microorganismos con morfología ligeramente curvada, gram negativos y oxidasa positivos. En cuanto a sus aspectos ecológicos son halófilos, por tanto suelen estar presentes en aguas marinas costeras y en el interior de moluscos, crustáceos y peces, y tienden a encontrarse en aguas cálidas. La patología que producen está asociada al consumo de mariscos,...

  11. Determination of several potential virulence factors in non-o1 Vibrio cholerae, Pseudomonas aeruginosa, faecal coliforms and streptococci isolated from Marrakesh groundwater.

    Science.gov (United States)

    Lamrani Alaoui, Hafsa; Oufdou, Khalid; Mezrioui, Nour-Eddine

    2010-01-01

    The dynamic, hemolytic and hemagglutination activities and the antibiotic resistance of non-O1 Vibrio cholerae, Pseudomonas aeruginosa, faecal coliforms (FC) and faecal streptococci (FS), isolated by standard membrane filtration methods from suburban and rural groundwater supplies, were carried out. Detectable non-O1 V. cholerae and P. aeruginosa was present in 81% and 88% of samples. The total occurrence of FC and FS during the period of study was 94%. The annual average densities of non-O1 V. cholerae were 4,903 MPN/100 mL. While, they were 206, 1,891 and 1,246 cfu/100 mL for P. aeruginosa, FC and FS respectively. Non-O1 V. cholerae strains had the highest percentage of hemolytic activities (alpha + beta) (71.29%), whereas 20.71% of FS, 16.88% of FC and 9.13% of P. aeruginosa strains produced hemolysin. Bacterial strains isolated were found to be adhesive, with percentages of 63.09%, 65.09%, 84.06% and 87.98% respectively for non-O1 V. cholerae, FS, FC and P. aeruginosa. As for antibiotic resistance, the overall resistance of non-O1 V. cholerae strains was 79%, whereas it was 100% for the other bacteria. Non-O1 V. cholerae resistance was expressed towards sulfamethoxazole (75%), streptomycin (62%) and cephalothin (60%). Obtained results indicated correlation between bacteriological pollution and their public health implications.

  12. Clinical isolates of Aeromonas veronii biovar veronii harbor a nonfunctional gene similar to the thermostable direct hemolysin-related hemolysin (trh) gene of Vibrio parahaemolyticus.

    Science.gov (United States)

    Raghunath, Pendru; Maiti, Biswajit; Shekar, Malathi; Karunasagar, Iddya; Karunasagar, Indrani

    2010-06-01

    Thermostable direct hemolysin-related hemolysin encoded by the trh gene is considered a major virulence factor in the pathogenesis of Vibrio parahaemolyticus infections. In this study, we report the presence of a trh homolog in three clinical isolates of Aeromonas veronii biovar veronii. The presence of a trh homolog in these strains of A. veronii was confirmed by PCR, followed by cloning, sequencing and colony hybridization using a digoxigenin-labelled probe. DNA sequence analysis revealed that the A. veronii trh gene had an identity of 99% and 84% to the trh1 and trh2 genes of V. parahaemolyticus, respectively. However, the expression of a trh-like gene in A. veronii could not be detected by reverse transcription PCR. Hence, the role of the gene product in the virulence of A. veronii strains is not clear. Further, these A. veronii isolates were negative for the ure gene encoding urease and the transposase gene by PCR. These genes are part of the trh gene cluster in V. parahaemolyticus. However, the presence of a trh homolog in a pathogen other than V. parahaemolyticus points to the fact that detection of the trh gene in stool samples, seafood enrichments or environmental samples does not always imply that trh-carrying V. parahaemolyticus are present.

  13. First isolation of Vibrio tapetis and an atypical strain of Aeromonas salmonicida from skin ulcerations in common dab (Limanda limanda) in the North Sea.

    Science.gov (United States)

    Vercauteren, M; De Swaef, E; Declercq, A; Bosseler, L; Gulla, S; Balboa, S; Romalde, J L; Devriese, L; Polet, H; Boyen, F; Chiers, K; Decostere, A

    2018-02-01

    Skin ulcerations rank amongst the most prevalent lesions affecting wild common dab (Limanda limanda) with an increase in prevalence of up to 3.5% in the Belgian part of the North Sea. A complex aetiology of these ulcerations is suspected, and many questions remain on the exact factors contributing to these lesions. To construct the aetiological spectrum of skin ulcerations in flatfish, a one-day monitoring campaign was undertaken in the North Sea. Fifteen fish presented with one or more ulcerations on the pigmented and/or non-pigmented side. Pathological features revealed various stages of ulcerations with loss of epidermal and dermal tissue, inflammatory infiltrates and degeneration of the myofibers bordering the ulceration, albeit in varying degrees. Upon bacteriological examination, pure cultures of Vibrio tapetis were retrieved in high numbers from five fish and of Aeromonas salmonicida in one fish. The V. tapetis isolates showed cross-reactivity with the sera against the representative strain of serotype O2 originating form a carpet-shell clam (Ruditapes descussatus). Moreover, the A. salmonicida isolates displayed a previously undescribed vapA gene sequence (A-layer type) with possible specificity towards common dab. Further research is necessary to pinpoint the exact role of these agents in the development of skin ulcerations in common dab. © 2017 John Wiley & Sons Ltd.

  14. Misidentification of Vibrio cholerae O155 isolated from imported shrimp as O serogroup O139 due to cross-agglutination with commercial O139 antisera

    DEFF Research Database (Denmark)

    Dalsgaard, A.; Mazur, J.; Dalsgaard, Inger

    2002-01-01

    Fish and shellfish products imported into Denmark are routinely analyzed for pathogenic Vibrio spp., particularly Vibrio cholerae, if products originate from subtropical or tropical areas. A V. cholerae strain that agglutinated commercial O139 antiserum but not the O1, Inaba, or Ogawa antisera...

  15. Enhanced Detection of Vibrio Cholerae in Oyster Homogenate Based on Centrifugal Removal of Inhibitory Agents

    Science.gov (United States)

    Alexander, Donita; DePaola, Angelo; Young, Ronald B.

    1998-01-01

    The disease cholera, caused by Vibrio cholerae, has been associated with consumption of contaminated seafood, including raw oysters. Detection of V. cholerae in foods typically involves blending the oysters, diluting the homogenate in alkaline peptone water (APW), overnight enrichment, and isolation on selective agar. Unfortunately, the oyster homogenate must be diluted to large volumes because lower dilutions inhibit the growth of V. cholerae. The goals of this study were to develop an alternative to large dilutions and to evaluate the basis for the inhibition observed in lower dilutions of oyster homogenates. Centrifugation of oyster homogenates at 10,000 x g for 15 min, followed by enrichment of the resulting pellet in APW, was found to eliminate the inhibition of V. cholerae growth. Inhibition appears not to be due to competing microflora but to a component(s) released when V. cholerae grows in the presence of oyster homogenate. The inhibitory component(s) kills the V. cholerae after the cell concentration reaches > 10(exp 8) cells/mL, rather than initially preventing their growth. The pH also declines from 8.0 to 5.5 during this period; however, the pH decline by itself appears not to cause V. cholerae death. Seven strains of V. cholerae (01 and non-01) and two strains of V. vulnificus were susceptible to the inhibitory agent(s). However, other Vibrio and non-Vibrio species tested were not inhibited by the oyster homogenates. Based on digestion of oyster homogenates with pronase, trypsin and lipase, the inhibitory reaction involves a protein(s). In a preliminary trial with oyster homogenate seeded with 1 cfu/g of V. cholerae, the modified centrifugation technique detected a slightly higher percentage of samples at a 1:10 dilution than the standard FDA Bacteriological Analytical Method (BAM) detected in uncentrifuged oyster homogenate at a 1:100 dilution. V. cholerae in seeded samples could also be detected more frequently by the modified centrifugation method

  16. Vibrios associated with red tides caused by Mesodinium rubrum.

    Science.gov (United States)

    Romalde, J L; Barja, J L; Toranzo, A E

    1990-11-01

    Vibrios were isolated from red tides caused by Mesodinium rubrum and also throughout the year in the Ria de Pontevedra, Spain. The isolates were grouped into 14 phena by numerical toxonomy. Strains associated with red tides were restricted to four phena: phena I and II were Vibrio alginolyticus, and phena III and IV were Vibrio tubiashii and Vibrio anguillarum, respectively. V. anguillarum-like strains (phena V through XI) predominated throughout the year outside the red tide areas. Cytotoxicity assays conducted in different poikilothermic and homoiothermic cell lines showed that cytotoxin production was not necessarily associated with the species selected during the red tides.

  17. Inhibitory Activity of Lactid Acid Bacteria Isolated from Tape Waterlily Seed to Enteric Pathogenic Bacteria (Vibrio cholera, Salmonella typhi, Shigella disentri, and E.coli and Its’ Susceptibility to Antibiotic, Bile Salt and Acidic Condition

    Directory of Open Access Journals (Sweden)

    Iin Khusnul Khotimah

    2012-03-01

    Full Text Available The aim of this research was to observe inhibitory activity of LAB isolated from tape waterlily seed to enteric pathogenic bacteria (Vibrio cholera, Salmonella typhi, Shigella disentri, E.coli ATCC 25922 and it’s susceptibility to antibiotic, in bile salt and under acidic condition. Microbia in the tape ( a fermented product of waterlily seed to showed were Streptococcus thermophilus (IKH-1, Pediococcus pentosaceus (IKH-2 and Leuconostoc mesentroides (IKH-8. Streptococcus thermophillus showed inhibition against the growth of Shigella disentri with inhibition zones 16,28 mm, but did not against the growth of V. Cholera, S. typhi, E.coli. Pediococcus pentosaceus inhibit Vibrio cholera, dan Salmonella thypi with inhibition zones 18,59 mm dan 7,91 mm. So that, Leuconostoc mesenteroides inhibit Salmonella thypi with zones inhibits average 8,25 mm. Chloramfenicol at 0.05 mg concentrations did not show inhibition against the growth of isolated Streptococcus thermophillus, Pediococcus pentosaceus and Leuconostoc mesentroides. These isolates could survive too in bile salt (2% and acidified media (pH 3.   Keyword : The tape of  waterlily seed, LAB, probiotic and enteric pathogenic   KEMAMPUAN PENGHAMBATAN BAKTERI ASAM LAKTAT DARI TAPE BIJI TERATAI TERHADAP PATOGENIK ENTERIK (VIBRIO CHOLERA, SALMONELLA THYPI, SHIGELLA DISENTRI, E. COLI, ANTIBIOTIK, KETAHANANNYA TERHADAP BILE SALT DAN ASAM   ABSTRAK   Penelitian ini bertujuan untuk menguji kemampuan penghambatan bakteri asam laktat yang diisolasi dari tape biji teratai terhadap patogenik enterik (Vibrio cholera, Salmonella thypi, Shigella disentri, E. Coli ATCC 25922, antibiotik, bile salt dan asam. Jenis bakteri yang diketahui tumbuh selama fermentasi tape biji teratai adalah Streptococcus thermopilus (IKH-1, Pediococcus pentosaceus(IKH-2, dan Leuconostoc mesentroides (IKH-8. Pengamatan terhadap uji penghambatan patogenik enterik (Vibrio cholera, Salmonella thypi, Shigella disentri, dan E. Coli ATCC

  18. Draft genome sequence of pathogenic bacteria Vibrio parahaemolyticus strain Ba94C2, associated with acute hepatopancreatic necrosis disease isolate from South America

    Directory of Open Access Journals (Sweden)

    Leda Restrepo

    2016-09-01

    Full Text Available Vibrio parahaemolyticus is a pathogenic bacteria which has been associated to the early mortality syndrome (EMS also known as hepatopancreatic necrosis disease (AHPND causing high mortality in shrimp farms. Pathogenic strains contain two homologous genes related to insecticidal toxin genes, PirA and PirB, these toxin genes are located on a plasmid contained within the bacteria. Genomic sequences have allowed the finding of two strains with a divergent structure related to the geographic region from where they were found. The isolates from the geographic collection of Southeast Asia and Mexico show variable regions on the plasmid genome, indicating that even though they are not alike they still conserve the toxin genes. In this paper, we report for the first time, a pathogenic V. parahaemolyticus strain in shrimp from South America that showed symptoms of AHPND. The genomic analysis revealed that this strain of V. parahaemolyticus found in South America appears to be more related to the Southeast Asia as compared to the Mexican strains. This finding is of major importance for the shrimp industry, especially in regards to the urgent need for disease control strategies to avoid large EMS outbreaks and economic loss, and to determine its dispersion in South America. The whole-genome shotgun project of V. parahaemolyticus strain Ba94C2 have been deposited at DDBJ/EMBL/GenBank under the accession PRJNA335761.

  19. Isolation and characterization of an antibacterium against Vibrio harveyi 11593 from a mixed pond with penaeus japonicus bate, portunus trituberculatus and ruditapes philippinarum in China

    Science.gov (United States)

    An, X. H.; Xu, Y. J.; Li, F. H.; Ma, G. Z.; Li, L. T.

    2016-08-01

    Aquaculture diseases, particularly Vibriosis, are becoming a pressing concern due to incurred aquacultural loss in China. Commercial antibiotics may be used to control it, but its increased antibiotic resistance to prominent pathogenic bacteria has become a prevalent problem nationwide, and a global threat to public health. Probiotics are recommended instead because they are healthy, environment-friendly, and capable of maintaining productivity. An antibacterium against Vibrio harveyi 11593 was isolated from a mariculture pond with shrimp, crabs, and shellfish in China. The bacterium, E14, has an inhibitory zone diameter (DIZ) of 24.5 ± 0.5 mm. The strain was identified as Bacillus pumilus based on morphological observation, conventional biochemical tests, and 16S rDNA sequence analysis. The gram-positiveand motile bacterium is around 1.10-1.32 pm × 0.67-0.83 μm in size. Optimized conditions for antimicrobial substance production of B. pumilus E14 require that it be cultured for 26 h at 28 °C, with an initial pH of 7.0 in 100 mL/500 mL LB with 3% NaCl. The B. pumilus E14 cultures were confirmed to be safe and efficacious and actually worked to protect the host animal in shrimp larvae (Penaeus chinensis) culture. The B. pumilus E14 obtained in this study strengthened the strain's defense against aquaculture disease and made a good candidate for an alternative probiotics and benefit to sustainability of aquaculture.

  20. Draft genome sequence of pathogenic bacteria Vibrio parahaemolyticus strain Ba94C2, associated with acute hepatopancreatic necrosis disease isolate from South America.

    Science.gov (United States)

    Restrepo, Leda; Bayot, Bonny; Betancourt, Irma; Pinzón, Andres

    2016-09-01

    Vibrio parahaemolyticus is a pathogenic bacteria which has been associated to the early mortality syndrome (EMS) also known as hepatopancreatic necrosis disease (AHPND) causing high mortality in shrimp farms. Pathogenic strains contain two homologous genes related to insecticidal toxin genes, PirA and PirB, these toxin genes are located on a plasmid contained within the bacteria. Genomic sequences have allowed the finding of two strains with a divergent structure related to the geographic region from where they were found. The isolates from the geographic collection of Southeast Asia and Mexico show variable regions on the plasmid genome, indicating that even though they are not alike they still conserve the toxin genes. In this paper, we report for the first time, a pathogenic V. parahaemolyticus strain in shrimp from South America that showed symptoms of AHPND. The genomic analysis revealed that this strain of V. parahaemolyticus found in South America appears to be more related to the Southeast Asia as compared to the Mexican strains. This finding is of major importance for the shrimp industry, especially in regards to the urgent need for disease control strategies to avoid large EMS outbreaks and economic loss, and to determine its dispersion in South America. The whole-genome shotgun project of V. parahaemolyticus strain Ba94C2 have been deposited at DDBJ/EMBL/GenBank under the accession PRJNA335761.

  1. Genome Sequence Analysis of Vibrio cholerae clinical isolates from 2013 in Mexico reveals the presence of the strain responsible for the 2010 Haiti outbreak.

    Science.gov (United States)

    Díaz-Quiñonez, José Alberto

    2017-01-01

    La primera semana de septiembre de 2013, el Sistema Nacional de Vigilancia Epidemiológica identificó dos casos de cólera en Ciudad de México. Los cultivos de ambas muestras se confirmaron como Vibrio cholerae serogrupo O1, serotipo Ogawa, biotipo El Tor. Los análisis iniciales por electroforesis por campos pulsados y por reacción en cadena de la polimerasa indicaron que ambas cepas eran similares, pero diferentes de las previamente reportadas en México. La semana siguiente se identificaron cuatro casos más en una comunidad del Estado de Hidalgo, ubicada a 121 kilómetros al noreste de Ciudad de México. Posteriormente se inició un brote de cólera en la región de La Huasteca. Los análisis genómicos de cuatro cepas obtenidas en este estudio confirmaron la presencia de las islas de patogenicidad VPI -1 y VPI-2, VSP-1 y VSP-2, y del elemento integrador SXT. La estructura genómica de los cuatro aislamientos fue similar a la de V. cholerae cepa 2010 EL-1786, identificada durante la epidemia en Haití en 2010. Este estudio pone de manifiesto que la epidemiología molecular es una herramienta muy poderosa para vigilar, prevenir y controlar enfermedades de importancia en salud pública en México. The first week of September 2013, the National Epidemiological Surveillance System identified two cases of cholera in Mexico City. The cultures of both samples were confirmed as Vibrio cholerae serogroup O1, serotype Ogawa, biotype El Tor. Initial analyses by pulsed-field gel electrophoresis and by polymerase chain reaction-amplification of the virulence genes, suggested that both strains were similar, but different from those previously reported in Mexico. The following week, four more cases were identified in a community in the state of Hidalgo, located 121 km northeast of Mexico City. Thereafter a cholera outbreak started in the region of La Huasteca. Genomic analyses of the strains obtained in this study confirmed the presence of pathogenicity islands VPI-1 and

  2. Antibiotic Susceptibility Patterns and Plasmid Profile of Vibrio ...

    African Journals Online (AJOL)

    32.14%) samples of Vibrio cholerae isolates recovered from water samples from Elele Community. All isolates showed a multiple resistance patterns to 7 antibiotics namely amoxicillin, cotrimoxazole, nitrofurantoin, gentamicin, tetracycline, ...

  3. Isolation of bioactive compound from marine seaweeds against fish pathogenic bacteria Vibrio alginolyticus (VA09 and characterisation by FTIR

    Directory of Open Access Journals (Sweden)

    Rajasekar Thirunavukkarasu

    2013-08-01

    Full Text Available Objective: Fresh marine seaweeds Gracilaria edulis, Gracillaria verrcosa, Acanthospora spicifera, Ulva facita, Ulva lacta (U. lacta, Kappaphycus spicifera, Sargassum ilicifolium, Sargassum wightii (S. wightii, Padina tetramatica and Padina gymonospora were collected from Mandapam (Rameshwaram, Tamil Nadu of South East coast of India and were screened for antibacterial activity. Methods: All the collected seaweeds were extracted by using five different solvent (methanol, isopropanol, acetone, chloroform, diethyl ether to study their extracts against fish pathogenic bacteria V. alginolyticus (VA09 purchased from MTCC. And minimum inhibition carried out by using Resazurin micro-titre assay. Crude extract of S. wightii analysied by FTIR. Results: The methanolic extract of S. wightii produced a maximum zone of inhibition (1.95±0.11 cm, isopropanol extract maximum inhibition was produced by S. wightii (1.93±0.78 cm, Acetone extract of Gracilaria verrcosa showed maximum zone of inhibition (1.36±0.05 cm, chloroform extract of S. wightii produced a maximum zone (1.56±0.25 cm and diethyl ether extract of S. wightii produced maximum zone of inhibition(1.86±0.11 cm. Based on the antibacterial activity S. wightii, U. lacta and Padina tetramatica showed best antibacterial activity against Vibrio harveyi. In this three seaweeds were taken for MIC study. The S. wightii methanolic extract, U. lacta diethyl ether extract and Padina tetramatica methanolic extract showed a higher MIC values, and despectively were 25 mg/mL, 50 mg/mL and 50 mg/mL. FTIR result showed that mostly phenolic compounds were present in the S. wightii. Conclusions: Based on the FTIR result S. wightii have high amount of phenolic compound. Phenolic compound have the good antimicrobial activity. The results clearly show that seaweed S. wightii is an interesting source for biologically active compounds that may be applied for prophylaxis and therapy of bacterial fish diseases and it should

  4. Comparative study on the antibiotic susceptibility and plasmid profiles of Vibrio alginolyticus strains isolated from four Tunisian marine biotopes.

    Science.gov (United States)

    Lajnef, Rim; Snoussi, Mejdi; Romalde, Jesús López; Nozha, Cohen; Hassen, Abdennaceur

    2012-12-01

    The antibiotic resistance patterns and the plasmids profiles of the predominant etiological agent responsible for vibriosis in Tunisia, V. alginolyticus were studied to contribute to control their spread in some Mediterranean aquaculture farms and seawater. The sixty-nine V. alginolyticus strains isolated from different marine Tunisian biotopes (bathing waters, aquaculture and conchylicole farms and a river connected to the seawater during the cold seasons) were multi-drug resistant with high resistance rate to ampicillin, kanamycin, doxycyclin, erythromycin, imipinem, and nalidixic acid. The multiple resistance index ranged from 0.3 to 0.7 for the isolates of Khenis, from 0.5 to 0.8 for those of Menzel Jmil, from 0.5 to 0.75 (Hergla) and from 0.3 to 0.7 for the isolates of Oued Soltane. The high value of antibiotic resistance index was recorded for the V. alginolyticus population isolated from the fish farm in Hergla (ARI = 0.672) followed by the population isolated from the conchylicole station of Menzel Jmil (ARI = 0.645). The results obtained by the MIC tests confirmed the resistance of the V. alginolyticus to ampicillin, erythromycin, kanamycin, cefotaxime, streptomycin and trimethoprim. Plasmids were found in 79.48 % of the strains analyzed and 30 different plasmid profiles were observed. The strains had a high difference in the size of plasmids varying between 0.5 and 45 kb. Our study reveals that the antibiotic-resistant bacteria are widespread in the aquaculture and conchylicole farm relatively to others strains isolated from seawater.

  5. Bacteriophage interactions with marine pathogenic Vibrios

    DEFF Research Database (Denmark)

    Kalatzis, Panagiotis

    Incidents of Vibrio-associated diseases in marine aquaculture are increasingly reported on a global scale, incited also by the world’s rising temperature. Administration of antibiotics has been the most commonly applied remedy used for facing vibriosis outbreaks, giving rise to concerns about...... pathogens. The combinatory administration of virulent bacteriophages φSt2 and φGrn1, isolated against Vibrio alginolyticus significantly reduced the Vibrio load in cultures of Artemia salina live prey, decreasing subsequently the risk of a vibriosis outbreak in the marine hatchery. During infection...... to studying the interactions between marine pathogenic Vibrio and their corresponding bacteriophages, while discussing the potential and limitations of phage therapy application in the biological control of vibriosis....

  6. Characterization of Vibrio cholerae O1 El Tor biotype variant clinical isolates from Bangladesh and Haiti, including a molecular genetic analysis of virulence genes.

    Science.gov (United States)

    Son, Mike S; Megli, Christina J; Kovacikova, Gabriela; Qadri, Firdausi; Taylor, Ronald K

    2011-11-01

    Vibrio cholerae serogroup O1, the causative agent of the diarrheal disease cholera, is divided into two biotypes: classical and El Tor. Both biotypes produce the major virulence factors toxin-coregulated pilus (TCP) and cholera toxin (CT). Although possessing genotypic and phenotypic differences, El Tor biotype strains displaying classical biotype traits have been reported and subsequently were dubbed El Tor variants. Of particular interest are reports of El Tor variants that produce various levels of CT, including levels typical of classical biotype strains. Here, we report the characterization of 10 clinical isolates from the International Centre for Diarrhoeal Disease Research, Bangladesh, and a representative strain from the 2010 Haiti cholera outbreak. We observed that all 11 strains produced increased CT (2- to 10-fold) compared to that of wild-type El Tor strains under in vitro inducing conditions, but they possessed various TcpA and ToxT expression profiles. Particularly, El Tor variant MQ1795, which produced the highest level of CT and very high levels of TcpA and ToxT, demonstrated hypervirulence compared to the virulence of El Tor wild-type strains in the infant mouse cholera model. Additional genotypic and phenotypic tests were conducted to characterize the variants, including an assessment of biotype-distinguishing characteristics. Notably, the sequencing of ctxB in some El Tor variants revealed two copies of classical ctxB, one per chromosome, contrary to previous reports that located ctxAB only on the large chromosome of El Tor biotype strains.

  7. Characterization of Vibrio cholerae O1 El Tor Biotype Variant Clinical Isolates from Bangladesh and Haiti, Including a Molecular Genetic Analysis of Virulence Genes ▿

    Science.gov (United States)

    Son, Mike S.; Megli, Christina J.; Kovacikova, Gabriela; Qadri, Firdausi; Taylor, Ronald K.

    2011-01-01

    Vibrio cholerae serogroup O1, the causative agent of the diarrheal disease cholera, is divided into two biotypes: classical and El Tor. Both biotypes produce the major virulence factors toxin-coregulated pilus (TCP) and cholera toxin (CT). Although possessing genotypic and phenotypic differences, El Tor biotype strains displaying classical biotype traits have been reported and subsequently were dubbed El Tor variants. Of particular interest are reports of El Tor variants that produce various levels of CT, including levels typical of classical biotype strains. Here, we report the characterization of 10 clinical isolates from the International Centre for Diarrhoeal Disease Research, Bangladesh, and a representative strain from the 2010 Haiti cholera outbreak. We observed that all 11 strains produced increased CT (2- to 10-fold) compared to that of wild-type El Tor strains under in vitro inducing conditions, but they possessed various TcpA and ToxT expression profiles. Particularly, El Tor variant MQ1795, which produced the highest level of CT and very high levels of TcpA and ToxT, demonstrated hypervirulence compared to the virulence of El Tor wild-type strains in the infant mouse cholera model. Additional genotypic and phenotypic tests were conducted to characterize the variants, including an assessment of biotype-distinguishing characteristics. Notably, the sequencing of ctxB in some El Tor variants revealed two copies of classical ctxB, one per chromosome, contrary to previous reports that located ctxAB only on the large chromosome of El Tor biotype strains. PMID:21880975

  8. Mortality event involving larvae of the carpet shell clam Ruditapes decussatus in a hatchery: isolation of the pathogen Vibrio tubiashii subsp. europaeus.

    Science.gov (United States)

    Dubert, J; Aranda-Burgos, J A; Ojea, J; Barja, J L; Prado, S

    2017-09-01

    Diseases caused by bacteria belonging to the genus Vibrio are a common, as yet unresolved, cause of mortality in shellfish hatcheries. In this study, we report the results of routine microbiological monitoring of larval cultures of the carpet shell clam Ruditapes decussatus in a hatchery in Galicia (NW Spain). Previous episodes of mortality with signs similar to those of vibriosis affecting other species in the installation indicated the possibility of bacterial infection and led to division of the culture at the early D-veliger larval stage. One batch was cultured under routine conditions, and the other was experimentally treated with antibiotic (chloramphenicol). Differences in larval survival were assessed, and culturable bacterial population in clams and sea water was evaluated, with particular attention given to vibrios. Severe mortalities were recorded from the first stages of culture onwards. The pathogen Vibrio tubiashii subsp. europaeus was detected in both batches, mainly associated with larvae. Moreover, initial detection of the pathogen in the eggs suggested the vertical transmission from broodstock as a possible source. Experimental use of antibiotic reduced the presence and diversity of vibrios in sea water, but proved inefficient in controlling vibrios associated with larvae from early stages and it did not stop mortalities. © 2017 John Wiley & Sons Ltd.

  9. Seasonal abundance of total and pathogenic Vibrio parahaemolyticus isolated from American oysters harvested in the Mandinga Lagoon System, Veracruz, Mexico: implications for food safety.

    Science.gov (United States)

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

    2014-07-01

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

  10. Reporte histórico: Primer Aislamiento de Vibrio cholera serogrupo O1 biovar El Tor serovar Inaba durante la epidemia de cólera en el Perú ‑ 1991 Historical report: first isolation of Vibrio cholera serogroup O1 biovar El Tor serovar Inaba during the cholerae epidemic in Perú ‑ 1991

    Directory of Open Access Journals (Sweden)

    Nora Bravo Cruz

    2011-03-01

    Full Text Available Hace 20 años apareció una enfermedad diarreica nueva en el Perú y el Laboratorio de Referencia de Enteropatógenos del Instituto Nacional de Salud, cumplió una labor destacada en el aislamiento e identificación rápida y oportuna del Vibrio cholerae. La enfermedad del cólera no se había presentado anteriormente, pero en la última semana de enero de 1991 se detectó un brote epidémico de diarrea aguda con deshidratación intensa y algunos casos de fallecidos. La epidemia afectó, al comienzo, varias localidades del litoral peruano. Equipos de trabajo de la Oficina General de Epidemiología y de los laboratorios del Instituto Nacional de Salud obtuvieron muestras fecales de pacientes con diarrea aguda procedentes de las ciudades de Chancay, Chimbote, Piura y algunos hospitales de Lima. Las muestras colectadas en el medio de transporte de Cary y Blair fueron procesadas en el Laboratorio Nacional de Referencia de Enteropatógenos (LANARE del Instituto Nacional de Salud. De todas las muestras se aisló e identificó Vibrio cholerae serogrupo O1 biovar El Tor serovar Inaba que mostró ser sensible a la tetraciclina y a otros antibióticos. Esta investigación confirmó el primer brote epidémico de cólera en el Perú.20 years ago, a new diarrheal disease was introduced in Peru and the Enteropathogens Reference Laboratory of the Instituto Nacional de Salud had an outstanding role in the isolation and rapid and timely identification of Vibrio cholerae. Cholera had not been seen before, but during the last week of January 1991 an outbreak of acute diarrhea was detected, presenting intense dehydration and some deaths. The epidemic affected, in the beginning, many locations of the peruvian coast. Some working teams of the General Office of Epidemiology and of the Instituto Nacional de Salud obtained fecal samples from patients with acute diarrhea coming from the cities of Chancay, Chimbote, Piura and some hospitals in Lima. The collected samples

  11. Highly diverse recombining populations of Vibrio cholerae and Vibrio parahaemolyticus in French Mediterranean coastal lagoons

    Science.gov (United States)

    Esteves, Kévin; Mosser, Thomas; Aujoulat, Fabien; Hervio-Heath, Dominique; Monfort, Patrick; Jumas-Bilak, Estelle

    2015-01-01

    Vibrio parahaemolyticus and Vibrio cholerae are ubiquitous to estuarine and marine environments. These two species found in Mediterranean coastal systems can induce infections in humans. Environmental isolates of V. cholerae (n = 109) and V. parahaemolyticus (n = 89) sampled at different dates, stations and water salinities were investigated for virulence genes and by a multilocus sequence-based analysis (MLSA). V. cholerae isolates were all ctxA negative and only one isolate of V. parahaemolyticus displayed trh2 gene. Most Sequence Types (ST) corresponded to unique ST isolated at one date or one station. Frequent recombination events were detected among different pathogenic species, V. parahaemolyticus, V. cholerae, Vibrio mimicus, and Vibrio metoecus. Recombination had a major impact on the diversification of lineages. The genetic diversity assessed by the number of ST/strain was higher in low salinity condition for V. parahaemolyticus and V. cholerae whereas the frequency of recombination events in V. cholerae was lower in low salinity condition. Mediterranean coastal lagoon systems housed V. cholerae and V. parahaemolyticus with genetic diversities equivalent to the worldwide diversity described so far. The presence of STs found in human infections as well as the frequency of recombination events in environmental vibrios populations could predict a potential epidemiological risk. PMID:26236294

  12. Antimicrobial susceptibilty of potentially pathogenic halophilic Vibrio ...

    African Journals Online (AJOL)

    Surveillance of antimicrobial resistance is indispensable for empirical treatment of infections and in preventing the spread of antimicrobial resistant microorganisms. This study is aimed at determining the antibiotic susceptibility of potentially pathogenic halophylic Vibrio species isolated in Lagos, Nigeria. Susceptibility ...

  13. Isolation of TDA-producing Phaeobacter strains from sea bass larval rearing units and their probiotic effect against pathogenic Vibrio spp. in Artemia cultures

    DEFF Research Database (Denmark)

    Grotkjær, Torben; Bentzon-Tilia, Mikkel; D'Alvise, Paul

    2016-01-01

    -DNA hybridization indicated that they could belong to a new species. The genomes contained genes involved in synthesis of the antibacterial compound tropodithietic acid (TDA), and its production was confirmed by UHPLC-TOFMS. The new Phaeobacter colonized live feed (Artemia) cultures and reduced Vibrio counts...... significantly, since they reached only 104 CFUmL-1, as opposed to 108 CFUmL-1 in non-Phaeobacter treated controls. Survival of V. anguillarum-challenged Artemia nauplii was enhanced by the presence of wild type Phaeobacter compared to challenged control cultures (89±1.0% vs 8±3.2%). In conclusion, TDA...

  14. Survival of Vibrio parahaemolyticus in cooked seafood at refrigeration temperatures.

    Science.gov (United States)

    Bradshaw, J G; Francis, D W; Twedt, R M

    1974-04-01

    The growth and survival of two strains of Vibrio parahaemolyticus isolated during food-borne gastroenteritis outbreaks in Japan and surface inoculated on cooked shrimp, shrimp with sauce, or cooked crab were tested at various refrigeration temperatures during a 48-h holding period. On cooked shrimp and crab, the vibrios grew well at 18.3 C, but their numbers declined gradually at 10 C and below. At 12.8 C, vibrios remained static for the most part. Thus, it appeared that 12.8 C was the borderline temperature for growth of the organism on cooked seafood. When cocktail sauce was added to surface-inoculated shrimp at a ratio of 2:1, the vibrio die-off rate was accelerated. In the shrimp and sauce few cells remained after 48 h, but in the sauce alone die-off was complete at 6 h.

  15. A rapid and reliable species-specific identification of clinical and environmental isolates of Vibrio cholerae using a three-test procedure and recA polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    A D Roozbehani

    2012-01-01

    Full Text Available Purpose: Vibrio cholerae, the cause of cholera, is one of the leading causes of morbidity and mortality in many developing countries. Most laboratories initially rely on biochemical tests for a presumptive identification of these strains, followed by a polymerase chain reaction (PCR-based method to confirm their identification. The aim of this study is to establish a rapid and reliable identification scheme for V. cholerae using a minimal, but highly specific number of biochemical tests and a PCR assay. Materials and Methods: We developed a species-specific PCR to identify V. cholerae, using a housekeeping gene recA, and used that to evaluate the sensitivity and specificity of 12 biochemical tests commonly used for screening and / or presumptive identification of V. cholerae in the clinical and environmental samples. Results: Here we introduced a combination of three biochemical tests, namely, sucrose fermentation, oxidase test, and growth in trypton broth containing 0% NaCl, as also the PCR of the recA gene, for rapid identification of V. cholerae isolates, with 100% sensitivity and specificity. The established method accurately identified a collection of 47 V. cholerae strains isolated from the clinical cases (n = 26 and surface waters (n = 21, while none of the 32 control strains belonging to different species were positive in this assay. Conclusion: The triple-test procedure introduced here is a simple and useful assay which can be adopted in cholera surveillance programs for efficient monitoring of V. cholerae in surface water and fecal samples.

  16. [Genomic variability of vibrio cholerae El Tor biovariant strains].

    Science.gov (United States)

    Smirnova, N I; Kostromitina, E A; Osin, A V; Kutyrev, V V

    2005-01-01

    The authors performed comparative analysis of the genomes of 145 clinical and environmental isolates of Vibrio cholerae El Tor biovariants using single locus and multiplex PCR. The study found that clinical strains isolated from patients with cholera formed a genetically homogenous group, where bacterial chromosome contained all the tested virulence genes, situated on mobile genetic elements that had been acquired by the pathogen at various stages of its evolution. Strains isolated from water ecosystems during interepidemic period were heterogeneous and formed three groups: a small number of virulent strains; non-toxigenic vibrio strains that, in the process of reductional variation in their new econiche, had only managed to maintain individual virulence genes; non-pathogenic "water" vibrios, whose chromosome contained only the genes from its core part, mobile genetic elements being optionally represented only by the persistence island. Molecular typing established genetic relations among V. cholerae strains under study.

  17. Vibrio parahaemolyticus produtores de urease isolados a partir de ostras (Crassostrea rizophorae coletadas in natura em restaurantes e mexilhões (Perna perna de banco natural Vibrio parahaemolyticus urease positive isolated from in natura oysters (Crassostrea rizophorae collected at restaurants and mussels (Perna perna harvested from natural habitat

    Directory of Open Access Journals (Sweden)

    Chistiane Soares Pereira

    2004-12-01

    Full Text Available A presença de Vibrio parahaemolyticus foi avaliada em 50 amostras de moluscos bivalves marinhos compostas por 40 amostras de ostras coletadas em 15 restaurantes do Rio de Janeiro e 10 amostras de mexilhões capturados de banco natural em Ponta de Itaipú - Niterói. Foram empregadas a técnica do Número Mais Provável (NMP para a enumeração de V. parahaemolyticus utilizando Caldo Glicosado Salgado com Teepol (GSTB e Água Peptonada Alcalina (APA com 3% de cloreto de sódio (NaCl. Paralelamente foi realizada técnica de enriquecimento em APA com 1 e 3% de NaCl. Decorrido o período de incubação de ambas as técnicas, foi realizado plaqueamento em ágar TCBS (Tiossulfato Citrato Bile Sacarose. Todas as cepas de V. parahaemolyticus isoladas através das duas técnicas foram testadas para o fenômeno de Kanagawa e, quanto à produção de urease. Do total de 141 cepas de V. parahaemolyticus isoladas, 62% revelaram-se urease positivas e, dentre estas, os sorotipos predominantes foram O10:K?, O11:K? e O3:K57 dentre o total de 24 sorotipos urease positivos identificados. Embora todas as cepas de V. parahaemolyticus tenham sido Kanagawa negativas, os resultados apontam elevada incidência desta espécie em ostras comercializadas em restaurantes.The presence of Vibrio parahaemolyticus were detected in 50 marine bivalve mollusks composed by 40 oysters samples collected at 15 restaurants in Rio de Janeiro City and 10 wild mussels' samples harvested in Ponta de Itaipu-Niterói. The Most Probable Number (MPN technique was employed for the enumeration of V. parahaemolyticus, using glucose salt teepol broth (GSTB and alcaline peptone water (APW with 3% NaCl. At the same time, the samples were submitted on direct plating with APW added 1 and 3% NaCl. Both techniques were followed by plating onto TCBS agar. All the V. parahaemolyticus strains isolated were tested for Kanagawa phenomenon and they were also tested for the presence of urease. A total of 141 V

  18. AI-2 quorum-sensing inhibitors affect the starvation response and reduce virulence in several Vibrio species, most likely by interfering with LuxPQ.

    Science.gov (United States)

    Brackman, Gilles; Celen, Shari; Baruah, Kartik; Bossier, Peter; Van Calenbergh, Serge; Nelis, Hans J; Coenye, Tom

    2009-12-01

    The increase of disease outbreaks caused by Vibrio species in aquatic organisms as well as in humans, together with the emergence of antibiotic resistance in Vibrio species, has led to a growing interest in alternative disease control measures. Quorum sensing (QS) is a mechanism for regulating microbial gene expression in a cell density-dependent way. While there is good evidence for the involvement of auto-inducer 2 (AI-2)-based interspecies QS in the control of virulence in multiple Vibrio species, only few inhibitors of this system are known. From the screening of a small panel of nucleoside analogues for their ability to disturb AI-2-based QS, an adenosine derivative with a p-methoxyphenylpropionamide moiety at C-3' emerged as a promising hit. Its mechanism of inhibition was elucidated by measuring the effect on bioluminescence in a series of Vibrio harveyi AI-2 QS mutants. Our results indicate that this compound, as well as a truncated analogue lacking the adenine base, block AI-2-based QS without interfering with bacterial growth. The active compounds affected neither the bioluminescence system as such nor the production of AI-2, but most likely interfered with the signal transduction pathway at the level of LuxPQ in V. harveyi. The most active nucleoside analogue (designated LMC-21) was found to reduce the Vibrio species starvation response, to affect biofilm formation in Vibrio anguillarum, Vibrio vulnificus and Vibrio cholerae, to reduce pigment and protease production in V. anguillarum, and to protect gnotobiotic Artemia from V. harveyi-induced mortality.

  19. Isolation and characterization of antimicrobial peptides derived from Bacillus subtilis E20-fermented soybean meal and its use for preventing Vibrio infection in shrimp aquaculture.

    Science.gov (United States)

    Cheng, Ann-Chang; Lin, Hsueh-Li; Shiu, Ya-Li; Tyan, Yu-Chang; Liu, Chun-Hung

    2017-08-01

    Bacillus subtilis E20-fermented soybean meal (FSBM) was found to produce antimicrobial peptides (AMPs) with great antimicrobial activity against Vibrio alginolyticus (VA) and V. parahaemolyticus (VP). Three AMPs were purified with a 5 kDa ultrafiltration, Sephadex G-15 column and reverse-phase high-performance liquid chromatography (RP-HPLC). The FSB-AMP, HTSKALLDMLKRLGK, identified by an RP-nano-ultrapure liquid chromatography (UPLC) electrospray ionization (ESI)-tandem mass spectroscopic (MS/MS) analysis exhibited the highest bactericidal activity against VA and VP compared to the others. The antimicrobial activity assessment indicated that FSB-AMP inhibited the growth of VA and VP with minimal inhibitory concentrations of 72.5 and 72.5 μM. Alterations in the morphology of VA were observed by scanning electronic microscopy, and membrane disruption of VA and VP was confirmed by fluorescent microscopy with propidium iodide staining. The FSB-AMP was then incorporated into the diet of white shrimp, Litopenaeus vannamei, and a protective effect in shrimp against VP infection was recorded as well as for shrimp fed a diet containing 15% fish meal replaced by B. subtilis E20-FSBM. Results demonstrated that B. subtilis E20-FSBM could be a biofunctional ingredient to prevent vibriosis in shrimp aquaculture. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. EROTYPE IDENTIFICATION OF VIBRIO CHOLERAE BACTERIAWHICH ISOLATED FROM ICE AMONGTUBE AND CUBE ICE TYPE IN FOOD AND BEVERAGES SELLER AT DENPASAR CITY, BALI

    Directory of Open Access Journals (Sweden)

    IGP Dhinarananta

    2014-01-01

    Full Text Available Cholera is a type of watery diarrhea with specific sign stool containing mucus which resembles rice water. Cholera caused by gram negative bacteria Vibrio cholerae (V.Cholerae. The transmissions of bacteria were through a contaminated food or water.Bali is an international tourism destination with tropical weather where ice is widelyused in food and beverage which bring a risk of cholera through a contaminated ice.Iceshave a risk of bacterial contamination whether from the making and the usage process.Type of ice that widely used were cube and tube ice which each of them have a differentin making and usage process. The purpose of this study is to obtain the contamination ofV.cholera in cube and tube ice. The method of this study is descriptive observationalstudy with quota sampling technique. Sample were obtained from a restaurants andstreet vendor which use a block and tube ice with total 10 sample and 5 for each type ofice.Sample then cultured in Alkaline Peptone Water(APW and Thiosulfate Citrate Bilesalt Sucrose(TCBS agar. Bacteriacolony then identified using a gram staining andLatex Serotyping. The result are 3 over 5 (60% sample of cube ice contaminated byV.cholera O1 Inaba serotype and 3 over 5 (60% sample of tube ice contaminated byV.cholera O1 Inaba serotype.

  1. Serotypes of Vibrio cholerae Non-O1 Isolated from Water Supplies for Human Consumption in Campeche, México and their Antibiotic Susceptibility Pattern

    Directory of Open Access Journals (Sweden)

    Isaac-Márquez Angélica P

    1998-01-01

    Full Text Available The presence of Vibrio cholerae non-O1 in water supplies for human consumption in the city of Campeche and rural locality of Bécal was investigated. V. cholerae non-O1 was detected in 5.9% of the samples obtained in deep pools of Campeche. Studies conducted in Bécal and neighbourhood of Morelos in Campeche indicated that collected samples harbored V. cholerae non-O1 in 31.5% and 8.7% respectively. There was a particular pattern of distribution of V. cholerae non-O1 serotypes among different studied regions. Accordingly, V. cholerae non-O1 serotype O14 predominated in the deep pools of Campeche and together with V. cholerae non-O1, O155 were preferentially founds in samples taken from intradomiciliary faucets in the neighbourhood of Morelos. Samples from Bécal predominantly presented the serotype O112. 60% and 53.8% of all studied strains of V. cholerae non-O1 proved to be resistant to ampicillin and carbenicillin. 3.1%, 7.7% and 6.2% presented resistant to doxycycline, trimethoprim-sulfamethoxazole and erythromycin respectively. The study showed the necessity of performing a strong epidemiologic surveillance for emergence and distribution of V. cholerae non-O1

  2. Virulence factors of non-O1 non-O139 Vibrio cholerae isolated in Córdoba, Argentina Factores de virulencia de Vibrio cholerae no-O1 no-O139 aislados en Córdoba, Argentina

    Directory of Open Access Journals (Sweden)

    C. Bidinost

    2004-12-01

    Full Text Available V. cholerae non-O1 non-O139 serogroups isolated from clinical and environmental sources in Córdoba, Argentina, were analyzed for the presence and expression of virulence genes. Most of the strains studied contained the genes toxR and hlyA, but lacked ctxA, zot, ace, tcpA and stn. The culture supernatants were tested for hemolytic and cytotoxic activity. The enterotoxic potential of the strains was studied in a rabbit ileal loop assay and their genetic profiles were compared by PFGE. The environmental strains varied in their virulence phenotype and showed no-clonal relationships. The clinical strains were highly enterotoxic, hemolytic, proteolytic and showed indistinguishable PFGE profiles, although they differed in their cytotoxic activity. This is the first description, using cell culture and “in vivo” studies, of the virulence properties of non-O1 non-O139 V. cholerae from Argentina.En este trabajo se analizó la presencia y expresión de genes de virulencia en V. cholerae no-O1 no-O139 de origen clínico y ambiental, aislados en Córdoba, Argentina. La mayoría de las cepas estudiadas contiene los genes toxR y hlyA, pero no ctxA, zot, ace, tcpA y stn. Se analizó la actividad hemolítica y citotóxica de estas cepas en los sobrenadantes de cultivo, así como su potencial enterotóxico en ensayos de asa ileal ligada de conejo. Además, los aislamientos fueron comparados por sus perfiles genéticos en PFGE. Las cepas del medio ambiente mostraron variación en su fenotipo de virulencia y no mostraron relación clonal. Las cepas clínicas fueron muy enterotóxicas, hemolíticas, proteolíticas y mostraron perfiles indistinguibles de PFGE, aunque mostraron diferencias en su actividad citotóxica. En este trabajo se describen por primera vez, utilizando ensayos de cultivo celular e “in vivo”, propiedades de virulencia de V. cholerae no-O1 no-O139 aislados en Argentina.

  3. Vibrio Parahaemolyticus: The Threat of Another Vibrio Acquiring Pandemic Potential

    Digital Repository Service at National Institute of Oceanography (India)

    Ramamurthy, T.; Nair, G.B.

    or indirectly via contaminated food and water. Food-borne Vibrio infections tend to occur more frequently in developed countries while transmission of Vibrio infections in developing countries is, by and large, water-borne. Further, the magnitude of food... and pandemics mainly due to poor water supply and personal hygiene. The other important and most common seafood-borne halophilic Vibrio is V. parahaemolyticus. Since its discovery in 1953 (Fujino et al., 1953), many aspects on this pathogen were explored...

  4. Vibrio na água e sedimento de viveiros de quatro fazendas de carcinicultura no estado do Ceará, Brasil

    Directory of Open Access Journals (Sweden)

    Regine Helena Silva dos Fernandes Vieira

    2010-12-01

    Full Text Available Foram realizadas 16 coletas, oito no período chuvoso e oito no período de estio, em quatro fazendas de carcinicultura do Estado do Ceará, nos estuários dos rios Jaguaribe (fazendas A e B e Acaraú (fazendas C e D, totalizando 32 amostras. O objetivo da pesquisa foi quantificar e identificar Vibrio spp. nas amostras de água e sedimento. Os valores máximos da Contagem Padrão em Placas (CPP de Vibrio spp. encontrados para as amostras de água, no período chuvoso, foram de 5.10³ UFC/mL est. e, para o sedimento, de 7,5.10³ UFC/g est. No período de estio, a CPP máxima para água foi de 4,35.10² UFC/mL est. e de 3,55.10³ UFC/g est. para as amostras de sedimento. Foram obtidos 36 isolados de Vibrio: Vibrio spp. (17, V. vulnificus B1(3; V. calviensis (2, V. cholerae (2, V. litoralis (2, V. metschnikovii (2, V. agarivorans (1, V. alginolyticus (1, V. campbellii (1, V. coralliilyticus (1, V. diazotrophicus (1, V. logei (1, V. mediterranei (1, V. vulnificus B2 (1. O conhecimento da presença de espécies, nunca anteriormente isoladas em viveiros de fazendas de carcinicultura, tais como o V. coralliilyticus, V. agarivorans, V. litoralis e V. calviensis são importantes para o monitoramento microbiológico contínuo desses ambientes.

  5. Retrospective genomic analysis of Vibrio cholerae O1 El Tor strains from different places in India reveals the presence of ctxB-7 allele found in Haitian isolates.

    Science.gov (United States)

    DE, R; Ramamurthy, T; Sarkar, B L; Mukhopadhyay, A K; Pazhani, G P; Sarkar, S; Dutta, S; Nair, G B

    2017-08-01

    A total of 45 strains of Vibrio cholerae O1 isolated from 10 different places in India where they were associated with cases of cholera between the years 2007 and 2008 were examined by molecular methods. With the help of phenotypic and genotypic tests the strains were confirmed to be O1 El Tor biotype strains with classical ctxB gene. Polymerase chain reaction (PCR) analysis by double - mismatch amplification mutation assay PCR showed 16 of these strains carried the ctxB-7 allele reported in Haitian strains. Sequencing of the ctxB gene in all the 45 strains revealed that in 16 strains the histidine at the 20th amino acid position had been replaced by asparagine and this single nucleotide polymorphism did not affect cholera toxin production as revealed by beads enzyme-linked immunosorbent assay. This study shows that the new ctxB gene sequence was circulating in different places in India. Seven representatives of these 45 strains analysed by pulsed - field gel electrophoresis showed four distinct Not I digested profiles showing that multiple clones were causing cholera in 2007 and 2008.

  6. Vibrio parahemolyticus septicaemia in a liver transplant patient: a case report

    Directory of Open Access Journals (Sweden)

    Fairweather Morgan G

    2011-05-01

    Full Text Available Abstract Introduction Vibrio parahemolyticus is the leading cause of vibrio-associated gastroenteritis in the United States of America, usually related to poor food handling; only rarely has it been reported to cause serious infections including sepsis and soft tissue infections. In contrast, Vibrio vulnificus is a well-known cause of septicaemia, especially in patients with cirrhosis. We present a patient with V. parahemolyticus sepsis who had an orthotic liver transplant in 2007 and was on immunosuppression for chronic rejection. Clinical suspicion driven by patient presentation, travel to Gulf of Mexico and soft tissue infection resulted in early diagnosis and institution of appropriate antibiotic therapy. Case presentation A 48 year old Latin American man with a history of chronic kidney disease, orthotic liver transplant in 2007 secondary to alcoholic end stage liver disease on immunosuppressants, and chronic rejection presented to the emergency department with fever, vomiting, abdominal pain, left lower extremity swelling and fluid filled blisters after a fishing trip in the Gulf of Mexico. Samples from the blister and blood grew V. parahemolyticus. The patient was successfully treated with ceftriaxone and ciprofloxacin. Conclusion Febrile patients with underlying liver disease and/or immunosuppression should be interviewed regarding recent travel to a coastal area and seafood ingestion. If this history is obtained, appropriate empiric antibiotics must be chosen. Patients with liver disease and/or immunosuppresion should be counselled to avoid eating raw or undercooked molluscan shellfish. People can prevent Vibrio sepsis and wound infections by proper cooking of seafood and avoiding exposure of open wounds to seawater or raw shellfish products.

  7. SEROTYPE IDENTIFICATION OF VIBRIO CHOLERAE ISOLATED FROM ICE THAT USE FOR MARINE PRODUCT PRESERVATIVE BETWEEN MODERN AND TRADITIONAL MARKET IN DENPASAR

    Directory of Open Access Journals (Sweden)

    I P Ananta WS

    2013-05-01

    Full Text Available 1024x768 Colera is a disease that cause by Vibrio cholerae (V. cholerae. V. cholerae is one of pathogen bacteria that can be gained from contaminated dietary sources. However, Indonesian people having indulgence in consuming seafood. They usually acquire the marine products at the nearby market. In the other side, unhygienic process in provision of raw materials possibly increase the contamination of V. cholerae. The purpose of this study are to obtain the contamination of V. cholerae on the marine products in Denpasar. This study using observational descriptive method with quotas sampling by taking fish ice preservative used by fish merchants in the modern market and traditional markets as the samples. Six samples taken in each location (traditional and modern market. Sample then cultured with medium alkaline peptone water ( APW continued with thiosulfate citrate bile salt sucrose ( TCBS . The result of the TCBS culture then continued by undergo grams staining and latex serotyping procedure to identify type V. vholerae. Every samples then noted and compared. The result on samples of modern market found 5 from 6 samples ( 83,33 % positive contains V. cholerae with Inaba serotype. Samples of traditional market found 4 from 6 sample (66,67% positive contains V. cholerae with inaba serotype and 1 of them (16,67% with Hikojima serotype. This study represent that contamination of V. cholerae are still high and potentially endanger people of Denpasar.   Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso

  8. Localization of immunodominant linear B-cell epitopes of Vibrio ...

    African Journals Online (AJOL)

    Outer membrane protein U (OmpU), an adhesion protein of Vibrio mimicus, is a good antigen, but its epitopes are still unclear. In order to locate the epitopes of OmpU protein, epitope prediction was performed using the amino acid sequence of OmpU protein of V. mimicus HX4 strain that was isolated from the diseased ...

  9. Genome sequence of the human pathogen Vibrio cholerae Amazonia.

    NARCIS (Netherlands)

    Thompson, C.C.; Marin, M.A.; Dias, G.M.; Dutilh, B.E.; Edwards, R.A.; Iida, T.; Thompson, F.L.; Vicente, A.C.

    2011-01-01

    Vibrio cholerae O1 Amazonia is a pathogen that was isolated from cholera-like diarrhea cases in at least two countries, Brazil and Ghana. Based on multilocus sequence analysis, this lineage belongs to a distinct profile compared to strains from El Tor and classical biotypes. The genomic analysis

  10. Vibrio Cholerae 01 Infections In Jos, Nigeria | Opajobi | African ...

    African Journals Online (AJOL)

    A study to determine the prevalence of Vibrio cholerae 01 in stool sample submitted for routine examination of enteric pathogens, as well as identify the serotypes and antibiogram of the isolates to commonly used antibiotics was undertaken. The survey involved the examination of 774 (763 stool and 11 rectal swabs) ...

  11. Pseudomonas piscicida kills vibrios by two distinct mechanisms

    Science.gov (United States)

    Pseudoalteromonas piscicida is a naturally-occurring marine bacterium which kills competing bacteria, including vibrios. In studies by Richards et al. (AEM00175-17), three strains of P. piscicida were isolated and characterized. Strains secreted proteolytic enzymes which likely killed competing or...

  12. Assessing the function of STAS domain protein SypA in Vibrio fischeri using a comparative analysis.

    Science.gov (United States)

    Thompson, Cecilia M; Visick, Karen L

    2015-01-01

    Colonization of the squid Euprymna scolopes by Vibrio fischeri requires biofilm formation dependent on the 18-gene symbiosis polysaccharide locus, syp. One key regulator, SypA, controls biofilm formation by an as-yet unknown mechanism; however, it is known that SypA itself is regulated by SypE. Biofilm-proficient strains form wrinkled colonies on solid media, while sypA mutants form biofilm-defective smooth colonies. To begin to understand the function of SypA, we used comparative analyses and mutagenesis approaches. sypA (and the syp locus) is conserved in other Vibrios, including two food-borne human pathogens, Vibrio vulnificus (rbdA) and Vibrio parahaemolyticus (sypA VP ). We found that both homologs could complement the biofilm defect of the V. fischeri sypA mutant, but their phenotypes varied depending on the biofilm-inducing conditions used. Furthermore, while SypAVP retained an ability to be regulated by SypE, RbdA was resistant to this control. To better understand SypA function, we examined the biofilm-promoting ability of a number of mutant SypA proteins with substitutions in conserved residues, and found many that were biofilm-defective. The most severe biofilm-defective phenotypes occurred when changes were made to a conserved stretch of amino acids within a predicted α-helix of SypA; we hypothesize that this region of SypA may interact with another protein to promote biofilm formation. Finally, we identified a residue required for negative control by SypE. Together, our data provide insights into the function of this key biofilm regulator and suggest that the SypA orthologs may play similar roles in their native Vibrio species.

  13. Spreading of Pandemic Vibrio parahaemolyticus O3:K6 and Its Serovariants: A Re-analysis of Strains Isolated from Multiple Studies

    Directory of Open Access Journals (Sweden)

    Dongsheng Han

    2017-05-01

    Full Text Available In China, V. parahaemolyticus has been a leading cause of foodborne outbreaks and bacterial infectious diarrhea since the 1990s, and most infections have been associated with the pandemic V. parahaemolyticus O3:K6 and its serovariants. However, a comprehensive overview of the sero-prevalence and genetic diversity of the pandemic V. parahaemolyticus clone in China is lacking. To compensate for this deficiency, pandemic isolates in both clinical and environmental Chinese samples collected from multiple studies were analyzed in this study. Surprisingly, as many as 27 clinical pandemic serovariants were identified and were widely distributed across nine coastal provinces and two inland provinces (Beijing and Sichuan. O3:K6, O4:K68, and O1:KUT represented the predominant clinical serovars. Only four environmental pandemic serovariants had previously been reported, and they were spread throughout Shanghai (O1:KUT, O3:K6, Jiangsu (O3:K6, O4:K48, Zhejiang (O3:K6, and Guangdong (O4:K9. Notably, 24 pandemic serovariants were detected within a short time frame (from 2006 to 2012. The pandemic isolates were divided into 15 sequence types (STs, 10 of which fell within clonal complex (CC 3. Only three STs (ST3, ST192, and ST305 were identified in environmental isolates. Substantial serotypic diversity was mainly observed among isolates within pandemic ST3, which comprised 21 combinations of O/K antigens. The pandemic O3:K6 serotype showed a high level of sequence diversity, which was shared by eight different STs (ST3, ST227, ST431, ST435, ST487, ST489, ST526, and ST672. Antimicrobial susceptibility testing revealed that most isolates shared similar antibiotic susceptibility profiles. They were resistant to ampicillin but sensitive to most other drugs that were tested. In conclusion, the high levels of serotypic and genetic diversity of the pandemic clone suggest that the involved regions are becoming important reservoirs for the emergence of novel pandemic

  14. Comparative assessment of Vibrio virulence in marine fish larvae

    DEFF Research Database (Denmark)

    Rønneseth, A.; Castillo, D.; D'Alvise, Paul

    2017-01-01

    Vibrionaceae infections are a major obstacle for marine larviculture; however, little is known about virulence differences of Vibrio strains. The virulence of Vibrio strains, mostly isolated from vibriosis outbreaks in farmed fish, was tested in larval challenge trials with cod (Gadus morhua...... effects on survival. Some Vibrio strains were pathogenic in all of the larva species, while some caused disease only in one of the species. Twenty-nine of the Vibrio anguillarum strains increased the mortality of larvae from at least one fish species; however, pathogenicity of the strains differed...... markedly. Other Vibrio species had no or less pronounced effects on larval mortalities. Iron uptake has been related to V. anguillarum virulence; however, the presence or absence of the plasmid pJM1 encoding anguibactin did not correlate with virulence. The genomes of V. anguillarum were compared (D...

  15. Demonstration and characterization of simultaneous production of a thermostable direct hemolysin (TDH/I) and a TDH-related hemolysin (TRHx) by a clinically isolated Vibrio parahaemolyticus strain, TH3766.

    Science.gov (United States)

    Xu, M; Iida, T; Yamamoto, K; Takarada, Y; Miwatani, T; Honda, T

    1994-01-01

    Simultaneous production of a thermostable direct hemolysin (TDH)-like toxin (TDHx) and a TDH-related hemolysin (TRH)-like toxin (TRHx) by a clinical isolate (strain TH3766) of Kanagawa phenomenon-positive Vibrio parahaemolyticus was demonstrated and characterized. The two hemolysins were differentially purified by column chromatography on hydroxyapatite and immunoaffinity columns. The molecular weight of the two hemolysins were estimated to be 23,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE). The purified TDHx was indistinguishable from the previously reported TDH/I (from strain TH012) but was different from the authentic TDH of a Kanagawa phenomenon-positive strain (T4750) physicochemically. The mobility of TRHx in nondenaturing PAGE differed from all the known TDHs and TRHs. The genes (tdhX and trhX) coding for TDHx and TRHx were cloned and sequenced. Homologies of nucleotide sequences of the coding regions between tdhX and tdhA (a gene for the authentic TDH) and between trhX and trh (a gene for the authentic TRH) were 98.1 and 99.1%, respectively, and homology between tdhX and trhX was 68.1%. At the amino acid level, TdhX was completely identical to TDH/I, although two base differences were found in the nucleotide sequences between tdhX and tdh/I. Two amino acid differences were observed between TrhX and Trh. Thus, these findings suggest that the TH3766 strain produces two types of hemolysins simultaneously. This is the first evidence that a strain of V. parahaemolyticus produces two types of toxins of the TDH-TRH family at the same time.

  16. Impact of milk fish farming in the tropics on potentially pathogenic vibrios.

    Science.gov (United States)

    Reichardt, W T; Reyes, J M; Pueblos, M J; Lluisma, A O

    2013-12-15

    Ratios of sucrose-negative to sucrose-positive vibrios on TCBS agar (suc-/suc+) indicate the abundance of potential human pathogenic non-cholera vibrios in coastal mariculture environments of the Lingayen Gulf (Philippines. In guts of adult maricultured milkfish (Chanos chanos) of suc- vibrios reached extreme peak values ranging between 2 and 545 million per g wet weight. Suc- vibrios outnumbered suc+ vibrios in anoxic sediments, too, and were rarely predominant in coastal waters or in oxidized sediments. Suc-/suc+ ratios in sediments increased toward the mariculture areas with distance from the open sea at decreasing redox potentials. There is circumstantial evidence that suc- vibrios can be dispersed from mariculture areas to adjacent environments including coral reefs. An immediate human health risk by pathogenic Vibrio species is discounted, since milkfish guts contained mainly members of the Enterovibrio group. A representative isolate of these contained proteolytic and other virulence factors, but no genes encoding toxins characteristic of clinical Vibrio species. Copyright © 2013 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

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

    2014-01-01

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

  18. Sigma E regulators control hemolytic activity and virulence in a shrimp pathogenic Vibrio harveyi.

    Directory of Open Access Journals (Sweden)

    Pimonsri Rattanama

    Full Text Available Members of the genus Vibrio are important marine and aquaculture pathogens. Hemolytic activity has been identified as a virulence factor in many pathogenic vibrios including V. cholerae, V. parahaemolyticus, V. alginolyticus, V. harveyi and V. vulnificus. We have used transposon mutagenesis to identify genes involved in the hemolytic activity of shrimp-pathogenic V. harveyi strain PSU3316. Out of 1,764 mutants screened, five mutants showed reduced hemolytic activity on sheep blood agar and exhibited virulence attenuation in shrimp (Litopenaeus vannamei. Mutants were identified by comparing transposon junction sequences to a draft of assembly of the PSU3316 genome. Surprisingly none of the disrupted open reading frames or gene neighborhoods contained genes annotated as hemolysins. The gene encoding RseB, a negative regulator of the sigma factor (σ(E, was interrupted in 2 out of 5 transposon mutants, in addition, the transcription factor CytR, a threonine synthetase, and an efflux-associated cytoplasmic protein were also identified. Knockout mutations introduced into the rpoE operon at the rseB gene exhibited low hemolytic activity in sheep blood agar, and were 3-to 7-fold attenuated for colonization in shrimp. Comparison of whole cell extracted proteins in the rseB mutant (PSU4030 to the wild-type by 2-D gel electrophoresis revealed 6 differentially expressed proteins, including two down-regulated porins (OmpC-like and OmpN and an upregulated protease (DegQ which have been associated with σ(E in other organisms. Our study is the first report linking hemolytic activity to the σ(E regulators in pathogenic Vibrio species and suggests expression of this virulence-linked phenotype is governed by multiple regulatory pathways within the V. harveyi.

  19. Genetic diversity of culturable Vibrio in an Australian blue mussel Mytilus galloprovincialis hatchery.

    Science.gov (United States)

    Kwan, Tzu Nin; Bolch, Christopher J S

    2015-09-17

    Bacillary necrosis associated with Vibrio species is the common cause of larval and spat mortality during commercial production of Australian blue mussel Mytilus galloprovincialis. A total of 87 randomly selected Vibrio isolates from various stages of rearing in a commercial mussel hatchery were characterised using partial sequences of the ATP synthase alpha subunit gene (atpA). The sequenced isolates represented 40 unique atpA genotypes, overwhelmingly dominated (98%) by V. splendidus group genotypes, with 1 V. harveyi group genotype also detected. The V. splendidus group sequences formed 5 moderately supported clusters allied with V. splendidus/V. lentus, V. atlanticus, V. tasmaniensis, V. cyclitrophicus and V. toranzoniae. All water sources showed considerable atpA gene diversity among Vibrio isolates, with 30 to 60% of unique isolates recovered from each source. Over half (53%) of Vibrio atpA genotypes were detected only once, and only 7 genotypes were recovered from multiple sources. Comparisons of phylogenetic diversity using UniFrac analysis showed that the culturable Vibrio community from intake, header, broodstock and larval tanks were phylogenetically similar, while spat tank communities were different. Culturable Vibrio associated with spat tank seawater differed in being dominated by V. toranzoniae-affiliated genotypes. The high diversity of V. splendidus group genotypes detected in this study reinforces the dynamic nature of microbial communities associated with hatchery culture and complicates our efforts to elucidate the role of V. splendidus group bacteria in vibriosis.

  20. Novel macrolide-resistance genes, mef(C) and mph(G), carried by plasmids from Vibrio and Photobacterium isolated from sediment and seawater of a coastal aquaculture site.

    Science.gov (United States)

    Nonaka, L; Maruyama, F; Suzuki, S; Masuda, M

    2015-07-01

    The aim of this study was to determine whether mef(C) and mph(G), originally found on the transferable multi-drug plasmid pAQU1 from Photobacterium damselae subsp. damselae isolated from seawater of a fish farm, are responsible for conferring macrolide resistance. Since these genes are localized head-to-tail on pAQU1 and only four nucleotides exist between them, the single- and combination-effect of these genes was examined. When mph(G) alone was introduced to Escherichia coli, the minimum inhibitory concentrations (MICs) against erythromycin, clarithromycin and azithromycin increased, whereas introduction of mef(C) alone did not influence macrolide susceptibility. Introduction of both mef(C) and mph(G) dramatically increased the MICs to the same three macrolides, i.e. >512 μg ml(-1) , >512 μg ml(-1) and 128 μg ml(-1) respectively. These results suggest that the macrolide phosphotransferase encoded by mph(G) is essential for macrolide resistance, while the efflux pump encoded by mef(C) is required for high-level macrolide resistance. The tandem-pair arrangements of the mef(C) and mph(G) genes were conserved on plasmids ranging in size from 240 to 350 kb of the 22 erythromycin-resistant strains belonging to Vibrio and Photobacterium obtained from the fish farm. Sixteen of 22 plasmids ranged in size from 300 to 350 kb. This is the first report of novel macrolide resistance genes originating from a marine bacterium. In this study, mef(C) and mph(G) were found to be novel macrolide-resistance genes, and this is the first report of macrolide-resistance genes originating from a marine bacterium. These genes may be responsible for previously reported cases of the emergence of erythromycin-resistant bacteria in aquaculture sites by an unknown mechanism. The introduction of the tandem arrangement of the mef(C) and mph(G) genes in Escherichia coli increased the MICs to erythromycin, clarithromycin and azithromycin, suggesting a novel mechanism conferring high

  1. Development of a More Sensitive and Specific Chromogenic Agar Medium for the Detection of Vibrio parahaemolyticus and Other Vibrio Species.

    Science.gov (United States)

    Yeung, Marie; Thorsen, Trevor

    2016-11-08

    Foodborne infections in the US caused by Vibrio species have shown an upward trend. In the genus Vibrio, V. parahaemolyticus is responsible for the majority of Vibrio-associated infections. Thus, accurate differentiation among Vibrio spp. and detection of V. parahaemolyticus is critically important to ensure the safety of our food supply. Although molecular techniques are increasingly common, culture-depending methods are still routinely done and they are considered standard methods in certain circumstances. Hence, a novel chromogenic agar medium was tested with the goal of providing a better method for isolation and differentiation of clinically relevant Vibrio spp. The protocol compared the sensitivity, specificity and detection limit for the detection of V. parahaemolyticus between the new chromogenic medium and a conventional medium. Various V. parahaemolyticus strains (n=22) representing diverse serotypes and source of origins were used. They were previously identified by Food and Drug Administration (FDA) and Centers for Disease Control and Prevention (CDC), and further verified in our laboratory by tlh-PCR. In at least four separate trials, these strains were inoculated on the chromogenic agar and thiosulfate-citrate-bile salts-sucrose (TCBS) agar, which is the recommended medium for culturing this species, followed by incubation at 35-37 °C for 24-96 hr. Three V. parahaemolyticus strains (13.6%) did not grow optimally on TCBS, nonetheless exhibited green colonies if there was growth. Two strains (9.1%) did not yield the expected cyan colonies on the chromogenic agar. Non-V. parahaemolyticus strains (n=32) were also tested to determine the specificity of the chromogenic agar. Among these strains, 31 did not grow or exhibited other colony morphologies. The mean recovery of V. parahaemolyticus on the chromogenic agar was ~96.4% relative to tryptic soy agar supplemented with 2% NaCl. In conclusion, the new chromogenic agar is an effective medium to detect V

  2. Marine Vibrio Species Produce the Volatile Organic Compound Acetone

    OpenAIRE

    Nemecek-Marshall, M.; Wojciechowski, C.; Kuzma, J.; Silver, G. M.; Fall, R.

    1995-01-01

    While screening aerobic, heterotrophic marine bacteria for production of volatile organic compounds, we found that a group of isolates produced substantial amounts of acetone. Acetone production was confirmed by gas chromatography, gas chromatography-mass spectrometry, and high-performance liquid chromatography. The major acetone producers were identified as nonclinical Vibrio species. Acetone production was maximal in the stationary phase of growth and was stimulated by addition of l-leucine...

  3. The iron-dependent regulator fur controls pheromone signaling systems and luminescence in the squid symbiont Vibrio fischeri ES114.

    Science.gov (United States)

    Septer, Alecia N; Lyell, Noreen L; Stabb, Eric V

    2013-03-01

    Bacteria often use pheromones to coordinate group behaviors in specific environments. While high cell density is required for pheromones to achieve stimulatory levels, environmental cues can also influence pheromone accumulation and signaling. For the squid symbiont Vibrio fischeri ES114, bioluminescence requires pheromone-mediated regulation, and this signaling is induced in the host to a greater extent than in culture, even at an equivalent cell density. Our goal is to better understand this environment-specific control over pheromone signaling and bioluminescence. Previous work with V. fischeri MJ1 showed that iron limitation induces luminescence, and we recently found that ES114 encounters a low-iron environment in its host. Here we show that ES114 induces luminescence at lower cell density and achieves brighter luminescence in low-iron media. This iron-dependent effect on luminescence required ferric uptake regulator (Fur), which we propose influences two pheromone signaling master regulators, LitR and LuxR. Genetic and bioinformatic analyses suggested that under low-iron conditions, Fur-mediated repression of litR is relieved, enabling more LitR to perform its established role as an activator of luxR. Interestingly, Fur may similarly control the LitR homolog SmcR of Vibrio vulnificus. These results reveal an intriguing regulatory link between low-iron conditions, which are often encountered in host tissues, and pheromone-dependent master regulators.

  4. Vibrio bivalvicida sp. nov., a novel larval pathogen for bivalve molluscs reared in a hatchery.

    Science.gov (United States)

    Dubert, Javier; Romalde, Jesús L; Prado, Susana; Barja, Juan L

    2016-02-01

    Three isolates were obtained from cultures of carpet shell clam (Ruditapes decussatus) reared in a bivalve hatchery (Galicia, NW Spain) from different sources: healthy broodstock, moribund larvae and the seawater corresponding to the larval tank. All isolates were studied by a polyphasic approach, including a phylogenetic analysis based on concatenated sequences of the five housekeeping genes ftsZ, gyrB, pyrH, recA and rpoA. The analysis supported their inclusion in the Orientalis clade of the genus Vibrio, and they formed a tight group separated from the closest relatives: Vibrio tubiashii subsp. europaensis, Vibrio tubiashii subsp. tubiashii and Vibrio orientalis. The percentages of genomic resemblance, including average nucleotide identity, DNA-DNA hybridization and in silico genome-to-genome comparison, between the type strain and the closest relatives were below values for species delineation and confirmed the taxonomic position of the new species, which could be differentiated from the related taxa on the basis of several phenotypic and chemotaxonomic features, including FAME and MALDI-TOF-MS. The pathogenicity of the new species was demonstrated in larvae of R. decussatus, Ruditapes philippinarum, Ostrea edulis and Donax trunculus. The results demonstrated that the strains analyzed represented a novel species in the Orientalis clade of the genus Vibrio, for which the name Vibrio bivalvicida sp. nov. is proposed, with 605(T) (= CECT 8855(T)=CAIM 1904(T)) designated as the type strain. Copyright © 2015 Elsevier GmbH. All rights reserved.

  5. Detection of Vibrio splendidus and related species in Chamelea gallina sampled in the Adriatic along the Abruzzi coastline

    Directory of Open Access Journals (Sweden)

    Marina Torresi

    2011-09-01

    Full Text Available Vibrio species are an important and widespread component of marine microbial communities. Some Vibrio strains are potentially pathogenic to marine vertebrates and invertebrates. The aim of this study was to identify vibrios, in particular Vibrio splendidus and related species, isolated from clams (Chamelea gallina collected along the coasts of the Abruzzi region from May to October 2007. The isolates obtained were phenotyped and classified as belonging to the genus Vibrio. The strains underwent biochemical testing in accordance with Alsina’s scheme for V. splendidus identification. Molecular analysis of the 16S-23S intergenic space region and recA gene was used to identify V. splendidus and related species. All the samples examined were found to contain halophylic Vibrio species, with V. alginolyticus, V. splendidus-related species and V. mediterranei most commonly found. A polymerase chain reaction of the 16S-23S intergenic space region and sequencing of the recA gene from isolates confirmed that phenotyping of Vibrio species is not sufficient to distinguish between different species. Differentiation of the highly related species among V. splendidus-related clusters remains an important issue. In this regard, our data suggests sequencing the recA genes was far more discriminatory than sequencing 16S rDNA for this purpose.

  6. Vibrio alginolyticus Associated Chronic Myringitis Acquired in Mediterranean Waters of Turkey

    Directory of Open Access Journals (Sweden)

    Burak Ekrem Citil

    2015-01-01

    Full Text Available Vibrio alginolyticus was originally classified as biotype 2 of Vibrio parahaemolyticus. Most clinical isolates are recovered from superficial wounds or the external ear infections. V. alginolyticus is acknowledged to be nearly nonpathogenic in humans. The reason for presence of V. alginolyticus’s virulence is uncertain. We describe a chronic myringitis case in a 47-year-old female due to V. alginolyticus. According to her anamnesis, it was detected that she had sea bathing history in Mugla Coast in Turkey. Pure isolation of V. alginolyticus was obtained from external auditory canal’s culture. Investigation and antimicrobial susceptibility of the isolate were performed by the automatized BD Phoenix system and Kirby-Bauer disk diffusion method, respectively. The bacteria were sensitive to all antibiotics. This case was presented to pay attention to Vibrio alginolyticus infections.

  7. Assessing the function of STAS domain protein SypA in Vibrio fischeri using a comparative analysis

    Directory of Open Access Journals (Sweden)

    Cecilia M Thompson

    2015-07-01

    Full Text Available Colonization of the squid Euprymna scolopes by Vibrio fischeri requires biofilm formation dependent on the 18-gene symbiosis polysaccharide locus, syp. One key regulator, SypA, controls biofilm formation by an as-yet unknown mechanism; however, it is known that SypA itself is regulated by SypE. Biofilm-proficient strains form wrinkled colonies on solid media, while sypA mutants form biofilm-defective smooth colonies. To begin to understand the function of SypA, we used comparative analyses and mutagenesis approaches. sypA (and the syp locus is conserved in other Vibrios, including two food-borne human pathogens, V. vulnificus (rbdA and V. parahaemolyticus (sypAVP. We found that both homologs could complement the biofilm defect of the V. fischeri sypA mutant, but their phenotypes varied depending on the biofilm-inducing conditions used. Furthermore, while SypAVP retained an ability to be regulated by SypE, RbdA was resistant to this control. To better understand SypA function, we examined the biofilm-promoting ability of a number of mutant SypA proteins with substitutions in conserved residues, and found many that were biofilm-defective. The most severe biofilm-defective phenotypes occurred when changes were made to a conserved stretch of amino acids within a predicted a-helix of SypA; we hypothesize that this region of SypA may interact with another protein to promote biofilm formation. Finally, we identified a residue required for negative control by SypE. Together, our data provide insights into the function of this key biofilm regulator and suggest that the SypA orthologs may play similar roles in their native Vibrio species.

  8. Occurrence of Vibrio species, beta-lactam resistant Vibrio species, and indicator bacteria in ballast and port waters of a tropical harbor.

    Science.gov (United States)

    Ng, Charmaine; Goh, Shin Giek; Saeidi, Nazanin; Gerhard, William A; Gunsch, Claudia K; Gin, Karina Yew Hoong

    2018-01-01

    Ballast water discharges are potential sources for the spread of invasive and pathogenic aquatic organisms. Ballast waters from six ships docked in the Port of Singapore were tested to determine if indictor organisms fell within proposed standards for ballast water discharge according to regulation D-2 of the Ballast Water Management Convention (BWMC) guidelines. Vibrio species were cultured on media supplemented with beta-lactam antibiotics to determine the presence of antibiotic resistant Vibrio species in the ballast waters of these vessels. Indicator organisms were quantified using culture media Colilert-18 and Enterolert in ballast waters of six ships docked in a tropical harbor, with uptake from different geographical locations. Of the six ships, one had ballast water originating from the Persian Gulf, another from the East China Sea, and four from the South China Sea. Two of the six ships which carried ballast waters from the East China Sea and the South China Sea did not meet the D-2 stipulated requirements of the Ballast Water Management Convention for indicator organisms with Enterococci values more than three times higher than the acceptable limit of 110 MPN/100mL), and Vibrio parahaemolyticus (2 to >110 MPN/100mL) were detected in at least one of six ballast water samples. Using thiosulfate-citrate-bile salts-sucrose agar (TCBS) supplemented with beta-lactam antibiotics (meropenem, ceftazidime), 11 different Vibrio species, exhibiting resistance to beta-lactam antibiotics were isolated; with Vibrio campbellii (44%) and Vibrio alginolyticus (15%) the most detected antibiotic resistant Vibrio species. A practical approach of prioritized screening of high-risk vessels should be conducted to ensure that the water quality meets D-2 standards prior to discharge. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Cholera outbreak caused by drug resistant Vibrio cholerae serogroup O1 biotype ElTor serotype Ogawa in Nepal; a cross-sectional study

    Directory of Open Access Journals (Sweden)

    Pappu Kumar Gupta

    2016-06-01

    Full Text Available Abstract Background Cholera is a major cause of mortality and morbidity in underdeveloped countries including Nepal. Recently drug resistance in Vibrio cholerae has become a serious problem mainly in developing countries. The main objectives of our study were to investigate the occurrence of Vibrio cholerae in stool samples from patients with watery diarrhea and to determine the antimicrobial susceptibility patterns of V. cholerae isolates. Methods A total of 116 stool samples from patients suffering from watery diarrhea during July to December 2012 were obtained from outbreak areas from all over Nepal. Alkaline peptone water and thiosulphate citrate bile salt sucrose agar (TCBS were used to isolate the Vibrio cholerae. The isolates were identified with the help of colony morphology, Gram’s staining, conventional biochemical testing, serotyping and biotyping. Antimicrobial susceptibility testing was performed by determining the minimum inhibitory concentration (MIC by agar dilution method. Results Vibrio cholerae was isolated from 26.72 % of total samples. All isolated Vibrio cholerae were confirmed to be Vibrio cholerae serogoup O1 biotype El Tor and serotype Ogawa. All isolates were resistant to ampicillin and cotrimoxazole. Twenty nine isolates were resistant toward two different classes of antibiotics, one strain was resistant to three different classes of antibiotics and one strain was resistant to four different classes of antibiotics. According to the definition of the multidrug resistant bacteria; 6.45 % of the strains of Vibrio cholerae were found to be multidrug resistant. Conclusions Cholera due to multidrug resistant Vibrio cholerae is also possible in Nepal. According to the antimicrobial susceptibility pattern of Vibrio cholerae in our study we recommend to use any antibiotics among tetracycline, doxycycline, levofloxacin, azithromycin, chloramphenicol and ciprofloxacin for preliminary treatment of cholera in Nepal.

  10. Vibrio sp. DSM 14379 pigment production--a competitive advantage in the environment?

    Science.gov (United States)

    Starič, Nejc; Danevčič, Tjaša; Stopar, David

    2010-10-01

    The ability to produce several antibacterial agents greatly increases the chance of producer's survival. In this study, red-pigmented Vibrio sp. DSM 14379 and Bacillus sp., both isolated from the same sampling volume from estuarine waters of the Northern Adriatic Sea, were grown in a co-culture. The antibacterial activity of the red pigment extract was tested on Bacillus sp. in microtiter plates. The MIC(50) for Bacillus sp. was estimated to be around 10⁻⁵ mg/L. The extract prepared form the nonpigmented mutant of Vibrio sp. had no antibacterial effect. The pigment production of Vibrio sp. was studied under different physicochemical conditions. There was no pigment production at high or low temperatures, high or low salt concentrations in peptone yeast extract (PYE) medium, low glucose concentration in mineral growth medium or high glucose concentration in PYE medium. This indicates that the red pigment production is a luxurious good that Vibrio sp. makes only under favorable conditions. The Malthusian fitness of Bacillus sp. in a co-culture with Vibrio sp. under optimal environmental conditions dropped from 4.0 to -7.6, which corresponds to three orders of magnitude decrease in the number of CFU relative to the monoculture. The nonpigmented mutant of Vibrio sp. in a co-culture with Bacillus sp. had a significant antibacterial activity. This result shows that studying antibacterial properties in isolation (i.e. pigment extract only) may not reveal full antibacterial potential of the bacterial strain. The red pigment is a redundant antibacterial agent of Vibrio sp.

  11. Essential oils of Nigella sativa protects Artemia from the pathogenic effect of Vibrio parahaemolyticus Dahv2.

    Science.gov (United States)

    Manju, Sivalingam; Malaikozhundan, Balasubramanian; Withyachumnarnkul, Boonsirm; Vaseeharan, Baskaralingam

    2016-05-01

    The anti-Vibrio activity of essential oils (EOs) of nine medicinal plants was tested against 28 Vibrio spp. isolated from diseased Fenneropenaeus indicus. EO of Nigella sativa exhibited anti-Vibrio activity against all Vibrio spp. and greater inhibition was noted for the isolate V2 which was identified as Vibrio parahaemolyticus Dahv2. Further, EO of N. sativa effectively inhibited V. parahaemolyticus Dahv2 with an inhibition zone of 23.9mm at 101.2μgml(-1). Moreover, EO of N. sativa revealed anti-biofilm activity at 101.2μgml(-1) against V. parahaemolyticus Dahv2 and inhibited the growth of V. parahaemolyticus Dahv2 at 100μgml(-1).In vivo experimental infection studies showed that the survival of Artemia spp. infected with V. parahaemolyticus Dahv2 at 1×10(3)cfuml(-1) was only 40%. However, the survival of Artemia spp. was significantly increased after challenge with 100μgml(-1) of EO of N. sativa. EO of N. sativa showed higher anti-oxidant potential and total phenol content than other EOs tested. The anti-oxidant activity of EO of N. sativa was highly correlated to their total phenolic contents (r=0.836, PArtemia spp. after experimental infection of V. parahaemolyticus Dahv2. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Identification of Vibrio spp. with a set of dichotomous keys.

    Science.gov (United States)

    Noguerola, I; Blanch, A R

    2008-07-01

    To define a binary biochemical key for the identification of all recognized Vibrio spp. A matrix of phenotypical results was developed based on the previous taxonomical studies and the first description manuscripts. A unification of results from various sources was also performed to integrate different taxonomical studies within the same data matrix. Established criteria for selecting the optimal set of tests yielded the highest discrimination, as well as the lowest number of tests. An initial identification key was defined using arginine dihydrolase, lysine decarboxylase and ornithine decarboxylase tests, as well as defining eight different clusters. This key leads each cluster to a secondary key for species identification. Most of Vibrio spp. presented an identification threshold of 100%. A new set of biochemical keys has been determined provides a scheme for the rapid identification of clinical and environmental species of Vibrio. No more than 14 are needed for even the most complicated identifications. This newly defined set of keys updates and improves similar findings published in previous studies. These biochemical keys are designed for use in routine applications, particularly in environmental and clinical studies involving a high number of isolates.

  13. Comparative Genomics of Vibrio cholerae from Haiti, Asia, and Africa

    Science.gov (United States)

    Reimer, Aleisha R.; Van Domselaar, Gary; Stroika, Steven; Walker, Matthew; Kent, Heather; Tarr, Cheryl; Talkington, Deborah; Rowe, Lori; Olsen-Rasmussen, Melissa; Frace, Michael; Sammons, Scott; Dahourou, Georges Anicet; Boncy, Jacques; Smith, Anthony M.; Mabon, Philip; Petkau, Aaron; Graham, Morag; Gilmour, Matthew W.

    2011-01-01

    Cholera was absent from the island of Hispaniola at least a century before an outbreak that began in Haiti in the fall of 2010. Pulsed-field gel electrophoresis (PFGE) analysis of clinical isolates from the Haiti outbreak and recent global travelers returning to the United States showed indistinguishable PFGE fingerprints. To better explore the genetic ancestry of the Haiti outbreak strain, we acquired 23 whole-genome Vibrio cholerae sequences: 9 isolates obtained in Haiti or the Dominican Republic, 12 PFGE pattern-matched isolates linked to Asia or Africa, and 2 nonmatched outliers from the Western Hemisphere. Phylogenies for whole-genome sequences and core genome single-nucleotide polymorphisms showed that the Haiti outbreak strain is genetically related to strains originating in India and Cameroon. However, because no identical genetic match was found among sequenced contemporary isolates, a definitive genetic origin for the outbreak in Haiti remains speculative. PMID:22099115

  14. Draft Genome Sequences of the Fish Pathogen Vibrio harveyi Strains VH2 and VH5

    DEFF Research Database (Denmark)

    Castillo, Daniel; D'Alvise, Paul; Middelboe, Mathias

    2015-01-01

    Vibrio harveyi is an important marine pathogen that is responsible for vibriosis outbreaks in cultured fish and invertebrates worldwide. Here, we announce the draft genome sequences of V. harveyi strains VH2 and VH5, isolated from farmed juvenile Seriola dumerili during outbreaks of vibriosis...

  15. Vibrio cholerae O1 from superficial water of the Tucunduba Stream, Brazilian Amazon

    Science.gov (United States)

    Sá, L.L.C.; Vale, E.R.V.; Garza, D.R.; Vicente, A.C.P.

    2012-01-01

    Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized. PMID:24031874

  16. Molecular phylogenetic analysis of Vibrio cholerae O1 El Tor strains ...

    Indian Academy of Sciences (India)

    Unknown

    all five ISR classes, could be successfully used to study phylogeny in this organism. [Ghatak A, Majumdar A and Ghosh R K 2005 Molecular phylogenetic analysis of Vibrio cholerae O1 El Tor strains isolated before, during and after the O139 outbreak based on the intergenomic heterogeneity of the 16S-23S rRNA intergenic ...

  17. Carbapenemase VCC-1-Producing Vibrio cholerae in Coastal Waters of Germany

    DEFF Research Database (Denmark)

    Hammerl, Jens A.; Jaeckel, Claudia; Bortolaia, Valeria

    2017-01-01

    During antimicrobial drug resistance testing for Vibrio spp. from coastal waters of Germany, we identified 4 nontoxigenic, carbapenem-resistant V. cholerae isolates. We used whole-genome sequencing to identify the carbapenemase gene bla(VCC-1). In addition, a molecular survey showed that more bla...

  18. The syp enhancer sequence plays a key role in transcriptional activation by the σ54-dependent response regulator SypG and in biofilm formation and host colonization by Vibrio fischeri.

    Science.gov (United States)

    Ray, Valerie A; Eddy, Justin L; Hussa, Elizabeth A; Misale, Michael; Visick, Karen L

    2013-12-01

    Biofilm formation by Vibrio fischeri is a complex process that requires multiple regulators. One such regulator, the NtrC-like response regulator SypG, controls biofilm formation and host colonization by V. fischeri via its impact on transcription of the symbiosis polysaccharide (syp) locus. SypG is predicted to activate syp transcription by binding to the syp enhancer (SE), a conserved sequence located upstream of four syp promoters. In this study, we performed an in-depth analysis of the sequences necessary for SypG to promote syp transcription and biofilm formation. We found that the SE sequence is necessary for SypG-mediated syp transcription, identified individual bases necessary for efficient activation, and determined that SypG is able to bind to syp promoter regions. We also identified SE sequences outside the syp locus and established that SypG recognizes these sequences as well. Finally, deletion of the SE sequence upstream of sypA led to defects in both biofilm formation and host colonization that could be restored by reintroducing the SE sequence into its native location in the chromosome. This work thus fills in critical gaps in knowledge of the Syp regulatory circuit by demonstrating a role for the SE sequence in SypG-dependent control of biofilm formation and host colonization and by identifying new putative regulon members. It may also provide useful insights into other bacteria, such as Vibrio vulnificus and Vibrio parahaemolyticus, that have syp-like loci and conserved SE sequences.

  19. Infection Vibrio sp. Bacteria on Kappaphycus Seaweed Varieties Brown and Green

    Science.gov (United States)

    Irmawati, Yuni; Sudirjo, Fien

    2017-10-01

    Disease in seaweed or ice-ice, until today is still a major problem in the cultivation of seaweed. Changes in extreme environmental conditions is a trigger factor of ice-ice, which can result in seaweed susceptible to infection with pathogenic microorganisms, such as bacteria Vibrio sp. This research aims to determine the bacteria Vibrio sp. infection in seaweed Kappaphycus varieties of brown and green. Vibrio sp. bacteria isolated in the infected seaweed thallus ice-ice, grown on TCBS media, purification, gram staining and biochemical tests. Vibrio sp. infected to seaweed Kappaphycus brown and green varieties in containers controlled by different density, 105 CFU/ml, 106 CFU/ml and 107CFU/ml. Observations were made to change clinical effect in thallus seaweed for 14 days of observation. The results obtained show that the levels of infection bacteria Vibrio sp. higher in seaweed Kappaphycus green varieties both in density 105 CFU/ml, 106 CFU/ml and 107CFU/ml, when compared with varieties brown.

  20. Complete genome sequence of a giant Vibrio phage ValKK3 infecting Vibrio alginolyticus

    Directory of Open Access Journals (Sweden)

    Tamrin M. Lal

    2016-06-01

    Full Text Available This paper describes the complete sequence of a giant lytic marine myophage, Vibrio phage ValKK3 that is specific to Vibrio alginolyticus ATCC® 17749™. Vibrio phage ValKK3 was subjected to whole genome sequencing on MiSeq sequencing platform and annotated using Blast2Go. The complete sequence of ValKK3 genome was deposited in DBBJ/EMBL/GenBank under accession number KP671755.

  1. Complete genome sequence of a giant Vibrio phage ValKK3 infecting Vibrio alginolyticus.

    Science.gov (United States)

    Lal, Tamrin M; Sano, Motohiko; Hatai, Kishio; Ransangan, Julian

    2016-06-01

    This paper describes the complete sequence of a giant lytic marine myophage, Vibrio phage ValKK3 that is specific to Vibrio alginolyticus ATCC(®) 17749™. Vibrio phage ValKK3 was subjected to whole genome sequencing on MiSeq sequencing platform and annotated using Blast2Go. The complete sequence of ValKK3 genome was deposited in DBBJ/EMBL/GenBank under accession number KP671755.

  2. Rapid discrimination of environmental Vibrio by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Eddabra, Rkia; Prévost, Gilles; Scheftel, Jean-Michel

    2012-04-20

    The aim of this study was to discriminate 30 Vibrio strains isolated from two wastewater treatment plants from Agadir, Morocco by two molecular typing methods, pulsed-field gel electrophoresis (PFGE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Out of the 30 strains of Vibrio examined in this study, 5 isolates could not be typed by PFGE and consistently appeared as a smear on the gel. In general, high genetic biodiversity among the Vibrio strains was found regardless to the isolation source. The results of MALDI TOF analysis show a high congruence of strain grouping demonstrating the accuracy and reliability of MALDI-TOF MS. Copyright © 2011 Elsevier GmbH. All rights reserved.

  3. Diversity and ecological structure of vibrios in benthic and pelagic habitats along a latitudinal gradient in the Southwest Atlantic Ocean

    Directory of Open Access Journals (Sweden)

    Luciane A. Chimetto Tonon

    2015-02-01

    Full Text Available We analyzed the diversity and population structure of the 775 Vibrio isolates from different locations of the southwestern Atlantic Ocean (SAO, including St. Peter and St. Paul Archipelago (SPSPA, Abrolhos Bank (AB and the St. Sebastian region (SS, between 2005 and 2010. In this study, 195 novel isolates, obtained from seawater and major benthic organisms (rhodoliths and corals, were compared with a collection of 580 isolates previously characterized (available at www.taxvibrio.lncc.br. The isolates were distributed in 8 major habitat spectra according to AdaptML analysis on the basis of pyrH phylogenetic reconstruction and ecological information, such as isolation source (i.e., corals: Madracis decactis, Mussismilia braziliensis, M. hispida, Phyllogorgia dilatata, Scolymia wellsi; zoanthids: Palythoa caribaeorum, P. variabilis and Zoanthus solanderi; fireworm: Hermodice carunculata; rhodolith; water and sediment and sampling site regions (SPSPA, AB and SS. Ecologically distinct groups were discerned through AdaptML, which finds phylogenetic groups that are significantly different in their spectra of habitat preferences. Some habitat spectra suggested ecological specialization, with habitat spectra 2, 3, and 4 corresponding to specialization on SPSPA, AB, and SS, respectively. This match between habitat and location may reflect a minor exchange of Vibrio populations between geographically isolated benthic systems. Moreover, we found several widespread Vibrio species predominantly from water column, and different populations of a single Vibrio species from H. carunculata in ecologically distinct groups (H-1 and H-8 respectively. On the other hand, AdaptML detected phylogenetic groups that are found in both the benthos and in open water. The ecological grouping observed suggests dispersal and connectivity between the benthic and pelagic systems in AB. This study is a first attempt to characterize the biogeographic distribution of vibrios in both

  4. Diversity and ecological structure of vibrios in benthic and pelagic habitats along a latitudinal gradient in the Southwest Atlantic Ocean

    Science.gov (United States)

    Silva, Bruno Sergio de O.; Moreira, Ana Paula B.; Valle, Cecilia; Alves, Nelson; Cavalcanti, Giselle; Garcia, Gizele; Lopes, Rubens M.; Francini-Filho, Ronaldo B.; de Moura, Rodrigo L.; Thompson, Cristiane C.

    2015-01-01

    We analyzed the diversity and population structure of the 775 Vibrio isolates from different locations of the southwestern Atlantic Ocean (SAO), including St. Peter and St. Paul Archipelago (SPSPA), Abrolhos Bank (AB) and the St. Sebastian region (SS), between 2005 and 2010. In this study, 195 novel isolates, obtained from seawater and major benthic organisms (rhodoliths and corals), were compared with a collection of 580 isolates previously characterized (available at www.taxvibrio.lncc.br). The isolates were distributed in 8 major habitat spectra according to AdaptML analysis on the basis of pyrH phylogenetic reconstruction and ecological information, such as isolation source (i.e., corals: Madracis decactis, Mussismilia braziliensis, M. hispida, Phyllogorgia dilatata, Scolymia wellsi; zoanthids: Palythoa caribaeorum, P. variabilis and Zoanthus solanderi; fireworm: Hermodice carunculata; rhodolith; water and sediment) and sampling site regions (SPSPA, AB and SS). Ecologically distinct groups were discerned through AdaptML, which finds phylogenetic groups that are significantly different in their spectra of habitat preferences. Some habitat spectra suggested ecological specialization, with habitat spectra 2, 3, and 4 corresponding to specialization on SPSPA, AB, and SS, respectively. This match between habitat and location may reflect a minor exchange of Vibrio populations between geographically isolated benthic systems. Moreover, we found several widespread Vibrio species predominantly from water column, and different populations of a single Vibrio species from H. carunculata in ecologically distinct groups (H-1 and H-8 respectively). On the other hand, AdaptML detected phylogenetic groups that are found in both the benthos and in open water. The ecological grouping observed suggests dispersal and connectivity between the benthic and pelagic systems in AB. This study is a first attempt to characterize the biogeographic distribution of vibrios in both seawater and

  5. Free living and plankton-associated vibrios: assessment in ballast water, harbor areas and coastal ecosystems in Brazil

    Directory of Open Access Journals (Sweden)

    Irma Nelly G. Rivera

    2013-01-01

    Full Text Available Ballast water is a major transport vector of exotic aquatic species and pathogenic microorganisms. The wide-ranging spread of toxigenic Vibrio cholerae O1 from harbor areas has been frequently ascribed to discharge of contaminated ballast water into eutrophic coastal environments, such as during the onset of the seventh cholera pandemic in South America in the early 1990s. To determine the microbiological hazards of ballast waters transported to Brazilian ports, we evaluated water and plankton samples taken from (i ballast water tanks of recently arrived ships, (ii port areas along the Brazilian coastline from ~1 to 32 oS and (iii three coastal areas in São Paulo State. Vibrio concentration and toxigenic V. cholerae O1 occurrence were analyzed. Plankton-associated vibrios were more abundant than free-living vibrios in all studied environments. Vibrio cholerae was found in 9.5% of ballast tanks and 24.2% of port samples, both as free-living and attached forms, and was absent off São Paulo State. Toxigenic V. cholerae O1 isolates (ctxA+, tcpA+, involved in cholera disease, were found in ballast water (2% and harbor (2% samples. These results confirm that ballast water is an important carrier of pathogenic organisms, and that monitoring of vibrios and other plankton-attached bacteria is of paramount importance in ballast water management programs.

  6. Vibrio diseases of marine fish populations

    Science.gov (United States)

    Colwell, R. R.; Grimes, D. J.

    1984-03-01

    Several Vibrio spp. cause disease in marine fish populations, both wild and cultured. The most common disease, vibriosis, is caused by V. anguillarum. However, increase in the intensity of mariculture, combined with continuing improvements in bacterial systematics, expands the list of Vibrio spp. that cause fish disease. The bacterial pathogens, species of fish affected, virulence mechanisms, and disease treatment and prevention are included as topics of emphasis in this review.

  7. Vibrio chromosomes share common history

    Directory of Open Access Journals (Sweden)

    Gevers Dirk

    2010-05-01

    Full Text Available Abstract Background While most gamma proteobacteria have a single circular chromosome, Vibrionales have two circular chromosomes. Horizontal gene transfer is common among Vibrios, and in light of this genetic mobility, it is an open question to what extent the two chromosomes themselves share a common history since their formation. Results Single copy genes from each chromosome (142 genes from chromosome I and 42 genes from chromosome II were identified from 19 sequenced Vibrionales genomes and their phylogenetic comparison suggests consistent phylogenies for each chromosome. Additionally, study of the gene organization and phylogeny of the respective origins of replication confirmed the shared history. Conclusions Thus, while elements within the chromosomes may have experienced significant genetic mobility, the backbones share a common history. This allows conclusions based on multilocus sequence analysis (MLSA for one chromosome to be applied equally to both chromosomes.

  8. Genome engineering in Vibrio cholerae

    DEFF Research Database (Denmark)

    Val, Marie-Eve; Skovgaard, Ole; Ducos-Galand, Magaly

    2012-01-01

    importance in public health, Vibrio cholerae, the causative agent of cholera, has become a preferred model to study bacteria with multipartite genomes. However, most in vivo studies in V. cholerae have been hampered by its genome architecture, as it is difficult to give phenotypes to a specific chromosome....... This difficulty was surmounted using a unique and powerful strategy based on massive rearrangement of prokaryotic genomes. We developed a site-specific recombination-based engineering tool, which allows targeted, oriented, and reciprocal DNA exchanges. Using this genetic tool, we obtained a panel of V. cholerae...... in V. cholerae and the general question concerning bacteria carrying circular chromosomes--by looking at the effect of chromosome size on topological issues. In this article, we show that Dam, RctB, and ParA2/ParB2 are strictly essential for chrII origin maintenance, and we formally demonstrate...

  9. Hatchery mortalities of larval oysters caused by Vibrio tubiashii and Vibrio coralliilyticus

    Science.gov (United States)

    Hatchery production of bivalve shellfish has been hampered by the occasional presence of opportunistic pathogens, particularly Vibrio coralliilyticus and Vibrio tubiashii. The present study reports the results of several avenues of research to better define these pathogens and the roles they play i...

  10. Directory of Open Access Journals (Sweden)

    Renata Albuquerque Costa

    2013-01-01

    Full Text Available INTRODUCTION: The present study aimed to survey the Vibrio microbiota of oysters (Crassostrea rhizophorae obtained from restaurants in Fortaleza, State of Ceará, Brazil, and to identify virulence factors. METHODS: The isolated vibrios were submitted to biochemical identification and were tested for hemolytic and urease activities. RESULTS: The isolated strains belonged to 13 species, with predominance of Vibrio mimicus. Of the strain isolates only from fresh samples, 20.5% and 2.8% showed hemolytic and urease activities, respectively. CONCLUSIONS: The findings support the little-publicized claim that Vibrio species other than V. parahaemolyticus and V. vulnificus can represent a health risk to public health.

  11. A Comprehensive Epidemiological Research for Clinical Vibrio parahaemolyticus in Shanghai

    Directory of Open Access Journals (Sweden)

    Huan Li

    2017-06-01

    Full Text Available Vibrio parahaemolyticus is one of the most important pathogen for seafood-borne gastroenteritis in Shanghai and the rest of the world. A total of 42 V. parahaemolyticus strains were isolated from 1900 fecal specimens collected from patients in Shanghai hospital presenting from January 2014 to December 2015. All isolates were evaluated for potential virulence factors [tdh, trh, and type three secretion system (T3SS genes], typed using multilocus sequence typing (MLST and screened for antimicrobial resistance phenotype and genotype. And for the first time, the relationship between virulence, genetic diversity and antimicrobial resistance of these isolates were identified. The results showed that 37 isolates carried the tdh gene (88.1% and only seven isolates were positive for the trh gene. The T3SS1 and T3SS2 genes were detected in all strains and only trh-positive isolates are also containing the T3SS2β genes. MLST analysis of the 42 Shanghai isolates identified 20 sequence types (STs with 16 novel STs and that these clinical V. parahaemolyticus strains showed high degrees of genetic diversity. All isolates expressed high levels of resistance against Ampicillin (100.0%, Streptomycin (100.0%, Cephazolin (92.9%, Kanamycin (92.8% and Amikacin (90.5%, and eight out of 38 resistance genes (SHV, tet(B, strA, qnrA, gryA, qnrB, sulI, sulII were detected in at least two isolates. This study confirms that antimicrobial resistance of clinical V. parahaemolyticus isolates is greater than those of environmental isolates. Furthermore, no clear correlation between antimicrobial resistance and virulence or genetic diversity was found in this study. These results add to epidemiological data of clinical V. parahaemolyticus isolates in Shanghai and highlight the need for additional mechanistic studies, especially antimicrobial resistance, to reduce the burden of disease caused by this pathogen in China.

  12. Phylogenetic Diversity of Vibrio cholerae Associated with Endemic Cholera in Mexico from 1991 to 2008

    Directory of Open Access Journals (Sweden)

    Seon Young Choi

    2016-03-01

    Full Text Available An outbreak of cholera occurred in 1991 in Mexico, where it had not been reported for more than a century and is now endemic. Vibrio cholerae O1 prototype El Tor and classical strains coexist with altered El Tor strains (1991 to 1997. Nontoxigenic (CTX− V. cholerae El Tor dominated toxigenic (CTX+ strains (2001 to 2003, but V. cholerae CTX+ variant El Tor was isolated during 2004 to 2008, outcompeting CTX−V. cholerae. Genomes of six Mexican V. cholerae O1 strains isolated during 1991 to 2008 were sequenced and compared with both contemporary and archived strains of V. cholerae. Three were CTX+ El Tor, two were CTX− El Tor, and the remaining strain was a CTX+ classical isolate. Whole-genome sequence analysis showed the six isolates belonged to five distinct phylogenetic clades. One CTX− isolate is ancestral to the 6th and 7th pandemic CTX+V. cholerae isolates. The other CTX− isolate joined with CTX− non-O1/O139 isolates from Haiti and seroconverted O1 isolates from Brazil and Amazonia. One CTX+ isolate was phylogenetically placed with the sixth pandemic classical clade and the V. cholerae O395 classical reference strain. Two CTX+ El Tor isolates possessing intact Vibrio seventh pandemic island II (VSP-II are related to hybrid El Tor isolates from Mozambique and Bangladesh. The third CTX+ El Tor isolate contained West African-South American (WASA recombination in VSP-II and showed relatedness to isolates from Peru and Brazil. Except for one isolate, all Mexican isolates lack SXT/R391 integrative conjugative elements (ICEs and sensitivity to selected antibiotics, with one isolate resistant to streptomycin. No isolates were related to contemporary isolates from Asia, Africa, or Haiti, indicating phylogenetic diversity.

  13. Characterization of Toxigenic Vibrio cholerae from Haiti, 2010–2011

    Science.gov (United States)

    Bopp, Cheryl; Tarr, Cheryl; Parsons, Michele B.; Dahourou, Georges; Freeman, Molly; Joyce, Kevin; Turnsek, Maryann; Garrett, Nancy; Humphrys, Michael; Gomez, Gerardo; Stroika, Steven; Boncy, Jacques; Ochieng, Benjamin; Oundo, Joseph; Klena, John; Smith, Anthony; Keddy, Karen; Gerner-Smidt, Peter

    2011-01-01

    In October 2010, the US Centers for Disease Control and Prevention received reports of cases of severe watery diarrhea in Haiti. The cause was confirmed to be toxigenic Vibrio cholerae, serogroup O1, serotype Ogawa, biotype El Tor. We characterized 122 isolates from Haiti and compared them with isolates from other countries. Antimicrobial drug susceptibility was tested by disk diffusion and broth microdilution. Analyses included identification of rstR and VC2346 genes, sequencing of ctxAB and tcpA genes, and pulsed-field gel electrophoresis with SfiI and NotI enzymes. All isolates were susceptible to doxycycline and azithromycin. One pulsed-field gel electrophoresis pattern predominated, and ctxB sequence of all isolates matched the B-7 allele. We identified the tcpETCIRS allele, which is also present in Bangladesh strain CIRS 101. These data show that the isolates from Haiti are clonally and genetically similar to isolates originating in Africa and southern Asia and that ctxB-7 and tcpETCIRS alleles are undergoing global dissemination. PMID:22099116

  14. Diversity of vibrios in the haemolymph of the spider crab Maja brachydactyla.

    Science.gov (United States)

    Gomez-Gil, B; Roque, A; Lacuesta, B; Rotllant, G

    2010-09-01

    The aim of this study was to characterise and identify vibrios isolated from the haemolymph of apparently healthy adult spider crabs (Maja brachydactyla) wild-caught in the Spanish localities of Galician coast and in the Canary Islands and also from captive animals held at IRTA's facilities in the Ebro Delta of Catalonia, north-west Spanish Mediterranean coast. A total of 277 bacterial isolates were obtained, and of these, 171 were characterised with rep-PCR, resulting electrophoretic bands were analysed and clusters formed. Identification of representative strains of each cluster was made by sequencing the 16S rRNA. Samples from animals caught in Galicia and captive at IRTA (around 15-18°C) rendered mostly species belonging to the Splendidus clade (72·2 and 76·6% respectively), commonly found in cold waters (below 20°C). Higher species diversity was found in the haemolymph of the captive animals. In the warmer Canary Islands waters (around 21°C), the diversity of vibrios is dominated by three clades, Harveyi (Vibrio core group, 39·3%), Orientalis (23·2%) and Splendidus (21·4%) with a species diversity that equals that of the colder captive animals. Differences in the vibrios populations were found in the haemolymph extracted from animals collected from the three localities. Potential new species were found, and their description is under way. As with other invertebrates, spider crabs also contain a diverse population of vibrios. These findings should help researchers to diagnose when a crab is infected. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

  15. Effects of Global Warming on Vibrio Ecology.

    Science.gov (United States)

    Vezzulli, Luigi; Pezzati, Elisabetta; Brettar, Ingrid; Höfle, Manfred; Pruzzo, Carla

    2015-06-01

    Vibrio-related infections are increasing worldwide both in humans and aquatic animals. Rise in global sea surface temperature (SST), which is approximately 1 °C higher now than 140 years ago and is one of the primary physical impacts of global warming, has been linked to such increases. In this chapter, major known effects of increasing SST on the biology and ecology of vibrios are described. They include the effects on bacterial growth rate, both in the field and in laboratory, culturability, expression of pathogenicity traits, and interactions with aquatic organisms and abiotic surfaces. Special emphasis is given to the effect of ocean warming on Vibrio interactions with zooplankters, which represent one of the most important aquatic reservoirs for these bacteria. The reported findings highlight the biocomplexity of the interactions between vibrios and their natural environment in a climate change scenario, posing the need for interdisciplinary studies to properly understand the connection between ocean warming and persistence and spread of vibrios in sea waters and the epidemiology of the diseases they cause.

  16. Genome analysis of the coral bleaching pathogen Vibrio shiloi.

    Science.gov (United States)

    Reshef, Leah; Ron, Eliora; Rosenberg, Eugene

    2008-08-01

    The past few decades have seen a world-wide increase in coral diseases, yet little is known about coral pathogens. In this study, techniques commonly used in pathogenomic research were applied to the coral pathogen Vibrio shiloi in order to identify genetic elements involved in its virulence. Suppressive subtractive hybridization was used to compare the gene content of V. shiloi to that of a closely related but non-pathogenic bacterium, Vibrio mediterranei, resulting in identification of several putative virulence factors and of three novel genomic islands. The entire genome of V. shiloi was further screened for genes related to previously characterized steps in infection: adhesion, superoxide dismutase production and toxin production. Exposure of pure cultures of V. shiloi to crushed coral tissues strongly affected the expression of seven genes encoding pili, zona occludins toxin (Zot) and a superoxide dismutase. Analysis of eight V. shiloi strains isolated in the last decade shows a shift of the natural population from strains carrying all three genomic islands to strains carrying none of them. This shift occurred following appearance of resistance in the coral Oculina patagonica to infection by V. shiloi. The relevance of these findings to the bleaching disease caused by V. shiloi is discussed.

  17. First study of Vibrios in larval cultures of pullet carpet shell clam (Venerupis corrugata in hatchery

    Directory of Open Access Journals (Sweden)

    Javier Dubert Pérez

    2014-07-01

    immediately following Prado et al. (2005, 2014. The bacteriological media used were Marine Agar, for heterotrophic marine bacteria, and Thiosulphate-Citrate-Bile-Sucrose, selective for vibrios. Bacterial counts were determined in all samples and expressed in colony forming unit (cfu per millilitre or gram. The different types of colony in the samples of larval cultures were isolated, purified and subjected to a basic phenotypic characterization. Further studies were carried out with the isolates that shared the main features of the genus Vibrio, including phenotypic tests and the genetic characterization by sequencing of 16S rRNA gene, as described by Prado et al. (2014. The counts of marine heterotrophic bacteria associated to broodstock gonad were similar in both groups (≈105 cfu/g. However, substantial differences were observed in the counts of presumptive vibrios, with higher values in the stock B (≈104 cfu/g than in the stock A (≈102 cfu/g. Interestingly, the larval batch A showed high bacterial counts in the initial samples, including presumptive vibrios, while the batch B began with lower numbers of total marine bacteria and vibrios. The survival rates at the end of the larval development were higher in batch B (40% B1, 32% B2 than in batch A (24% A1, 27% A2. The use of antibiotic did not imply a significant enhancement of the survival. The total counts in the seawater from the culture tanks were similar. The presumptive vibrios were found mainly associated to larvae, regardless the batch or the use of antibiotic. The differences on the composition of bacterial populations will be analysed, focused on the identification of the presumptive vibrios. Preliminary results showed the dominance of species within the Splendidus clade of the genus Vibrio, as expected taking into account that its members have been isolated from marine environments and many of them from bivalves. Due to the high phenotypic diversity within this clade and the difficult to differentiate

  18. DETECTION OF VIRULENCE GENES IN ENVIRONMENTAL STRAINS OF Vibrio cholerae FROM ESTUARIES IN NORTHEASTERN BRAZIL

    Directory of Open Access Journals (Sweden)

    Francisca Gleire Rodrigues de Menezes

    2014-09-01

    Full Text Available The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS, and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

  19. Detection of virulence genes in environmental strains of Vibrio cholerae from estuaries in northeastern Brazil.

    Science.gov (United States)

    Menezes, Francisca Gleire Rodrigues de; Neves, Soraya da Silva; Sousa, Oscarina Viana de; Vila-Nova, Candida Machado Vieira Maia; Maggioni, Rodrigo; Theophilo, Grace Nazareth Diogo; Hofer, Ernesto; Vieira, Regine Helena Silva dos Fernandes

    2014-01-01

    The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

  20. DETECTION OF VIRULENCE GENES IN ENVIRONMENTAL STRAINS OF Vibrio cholerae FROM ESTUARIES IN NORTHEASTERN BRAZIL

    Science.gov (United States)

    de Menezes, Francisca Gleire Rodrigues; Neves, Soraya da Silva; de Sousa, Oscarina Viana; Vila-Nova, Candida Machado Vieira Maia; Maggioni, Rodrigo; Theophilo, Grace Nazareth Diogo; Hofer, Ernesto; Vieira, Regine Helena Silva dos Fernandes

    2014-01-01

    The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes. PMID:25229224

  1. Production of acylated homoserine lactones by different serotypes of Vibrio anguillarum both in culture and during infection of rainbow trout

    DEFF Research Database (Denmark)

    Buch, Christiane; Sigh, Jens; Nielsen, John

    2003-01-01

    Onehundred and forty-eight out of onehundred and fifty strains of Vibrio anguillarum isolated from vibriosis in Danish marine aquaculture produced bacterial communication signals, acylated homoserine lactones, eliciting a response in the Agrobacterium tumefaciens (pZLR4) monitoring system. One....... anguillarum strains and that no clear pattern relating AHL production to disease or virulence appear....

  2. Antioxidative metabolites synthesized by marine pigmented vibrio sp. and its protection on oxidative deterioration of membrane lipids

    Digital Repository Service at National Institute of Oceanography (India)

    Pawar, R.T.; Mohandass, C.; Dastager, S.G.; Kolekar, Y.M.; Malwankar, R.

    Bacterial strain Vibrio sp. (PIGB 184) isolated from water samples of the Arabian Sea and identified through 16S rRNA demonstrated the production of pigmentary antioxidants with higher ABTS activities 90.9±0.42 % in comparison with the standard...

  3. Acetone Formation in the Vibrio Family: a New Pathway for Bacterial Leucine Catabolism

    Science.gov (United States)

    Nemecek-Marshall, Michele; Wojciechowski, Cheryl; Wagner, William P.; Fall, Ray

    1999-01-01

    There is current interest in biological sources of acetone, a volatile organic compound that impacts atmospheric chemistry. Here, we determined that leucine-dependent acetone formation is widespread in the Vibrionaceae. Sixteen Vibrio isolates, two Listonella species, and two Photobacterium angustum isolates produced acetone in the presence of l-leucine. Shewanella isolates produced much less acetone. Growth of Vibrio splendidus and P. angustum in a fermentor with controlled aeration revealed that acetone was produced after a lag in late logarithmic or stationary phase of growth, depending on the medium, and was not derived from acetoacetate by nonenzymatic decarboxylation in the medium. l-Leucine, but not d-leucine, was converted to acetone with a stoichiometry of approximately 0.61 mol of acetone per mol of l-leucine. Testing various potential leucine catabolites as precursors of acetone showed that only α-ketoisocaproate was efficiently converted by whole cells to acetone. Acetone production was blocked by a nitrogen atmosphere but not by electron transport inhibitors, suggesting that an oxygen-dependent reaction is required for leucine catabolism. Metabolic labeling with deuterated (isopropyl-d7)-l-leucine revealed that the isopropyl carbons give rise to acetone with full retention of deuterium in each methyl group. These results suggest the operation of a new catabolic pathway for leucine in vibrios that is distinct from the 3-hydroxy-3-methylglutaryl-coenzyme A pathway seen in pseudomonads. PMID:10601206

  4. Vibrio population structure - Genetic and population structure analysis of clinical and environmental Vibrio parahaemolyticus strains

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Vibrio parahaemolyticus (Vp) is a marine bacterium capable of causing severe gastroenteritis in humans, usually through the consumption of raw shellfish. Before...

  5. Vibrio ecology - Identifying Environmental Determinants Favorable for the Presence and Transmission of Pathogenic Vibrios

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — In a tri-coastal collaborative study, the population densities of vibrios are being determined in the Mississippi Sound, Puget Sound, Chesapeake Bay, and Timbalier...

  6. Entry of Vibrio harveyi and Vibrio fischeri into the viable but nonculturable state

    Digital Repository Service at National Institute of Oceanography (India)

    Ramaiah; Ravel, J.; Straube, W.L.; Hill, R.T.; Colwell, R.R.

    Physical responses of marine luminous bacteria, Vibrio harveyi (ATCC 14216) and V. fischeri (UM1373) to nutrient-limited normal strength (35 ppt iso-osmolarity) and low (10 ppt hypo-osmolarity) salinity conditions were determined. Plate counts...

  7. Free-Living and Plankton-Associated Vibrios: Assessment in Ballast Water, Harbor Areas, and Coastal Ecosystems in Brazil

    Science.gov (United States)

    Rivera, Irma N. G.; Souza, Keili M. C.; Souza, Claudiana P.; Lopes, Rubens M.

    2013-01-01

    Ballast water (BW) is a major transport vector of exotic aquatic species and pathogenic microorganisms. The wide-ranging spread of toxigenic Vibrio cholerae O1 from harbor areas has been frequently ascribed to discharge of contaminated BW into eutrophic coastal environments, such as during the onset of the seventh cholera pandemic in South America in the early 1990s. To determine the microbiological hazards of BWs transported to Brazilian ports, we evaluated water and plankton samples taken from (i) BW tanks of recently arrived ships, (ii) port areas along the Brazilian coastline from ∼1 to 32°S and (iii) three coastal areas in São Paulo State. Vibrio concentration and toxigenic V. cholerae O1 occurrence were analyzed. Plankton-associated vibrios were more abundant than free-living vibrios in all studied environments. V. cholerae was found in 9.5% of ballast tanks and 24.2% of port samples, both as free-living and attached forms and, apart from the Santos harbor, was absent off São Paulo State. Toxigenic V. cholerae O1 isolates (ctxA+, tcpA+), involved in cholera disease, were found in BW (2%) and harbor (2%) samples. These results confirm that BW is an important carrier of pathogenic organisms, and that monitoring of vibrios and other plankton-attached bacteria is of paramount importance in BW management programs. PMID:23335920

  8. Free-living and plankton-associated vibrios: assessment in ballast water, harbor areas, and coastal ecosystems in Brazil.

    Science.gov (United States)

    Rivera, Irma N G; Souza, Keili M C; Souza, Claudiana P; Lopes, Rubens M

    2012-01-01

    Ballast water (BW) is a major transport vector of exotic aquatic species and pathogenic microorganisms. The wide-ranging spread of toxigenic Vibrio cholerae O1 from harbor areas has been frequently ascribed to discharge of contaminated BW into eutrophic coastal environments, such as during the onset of the seventh cholera pandemic in South America in the early 1990s. To determine the microbiological hazards of BWs transported to Brazilian ports, we evaluated water and plankton samples taken from (i) BW tanks of recently arrived ships, (ii) port areas along the Brazilian coastline from ∼1 to 32°S and (iii) three coastal areas in São Paulo State. Vibrio concentration and toxigenic V. cholerae O1 occurrence were analyzed. Plankton-associated vibrios were more abundant than free-living vibrios in all studied environments. V. cholerae was found in 9.5% of ballast tanks and 24.2% of port samples, both as free-living and attached forms and, apart from the Santos harbor, was absent off São Paulo State. Toxigenic V. cholerae O1 isolates (ctxA(+), tcpA(+)), involved in cholera disease, were found in BW (2%) and harbor (2%) samples. These results confirm that BW is an important carrier of pathogenic organisms, and that monitoring of vibrios and other plankton-attached bacteria is of paramount importance in BW management programs.

  9. Environmental reservoirs and mechanisms of persistence of Vibrio cholerae

    Science.gov (United States)

    Lutz, Carla; Erken, Martina; Noorian, Parisa; Sun, Shuyang; McDougald, Diane

    2013-01-01

    It is now well accepted that Vibrio cholerae, the causative agent of the water-borne disease cholera, is acquired from environmental sources where it persists between outbreaks of the disease. Recent advances in molecular technology have demonstrated that this bacterium can be detected in areas where it has not previously been isolated, indicating a much broader, global distribution of this bacterium outside of endemic regions. The environmental persistence of V. cholerae in the aquatic environment can be attributed to multiple intra- and interspecific strategies such as responsive gene regulation and biofilm formation on biotic and abiotic surfaces, as well as interactions with a multitude of other organisms. This review will discuss some of the mechanisms that enable the persistence of this bacterium in the environment. In particular, we will discuss how V. cholerae can survive stressors such as starvation, temperature, and salinity fluctuations as well as how the organism persists under constant predation by heterotrophic protists. PMID:24379807

  10. The Vibrio cholerae cytolysin promotes chloride secretion from intact human intestinal mucosa.

    Directory of Open Access Journals (Sweden)

    Lucantonio Debellis

    Full Text Available BACKGROUND: The pathogenicity of the Vibrio cholerae strains belonging to serogroup O1 and O139 is due to the production of virulence factors such as cholera toxin (CT and the toxin-coregulated pilus (TCP. The remaining serogroups, which mostly lack CT and TCP, are more frequently isolated from aquatic environmental sources than from clinical samples; nevertheless, these strains have been reported to cause human disease, such as sporadic outbreaks of watery diarrhoea and inflammatory enterocolitis. This evidence suggested the possibility that other virulence factor(s than cholera toxin might be crucial in the pathogenesis of Vibrio cholerae-induced diarrhoea, but their nature remains unknown. VCC, the hemolysin produced by virtually all Vibrio cholerae strains, has been proposed as a possible candidate, though a clear-cut demonstration attesting VCC as crucial in the pathogenesis of Vibrio cholerae-induced diarrhoea is still lacking. METHODOLOGY/PRINCIPAL FINDINGS: Electrophysiological parameters and paracellular permeability of stripped human healthy colon tissues, obtained at subtotal colectomy, mounted in Ussing chamber were studied in the presence or absence of VCC purified from culture supernatants of V. cholerae O1 El Tor strain. Short circuit current (I(SC and transepithelial resistance (R(T were measured by a computerized voltage clamp system. The exposure of sigmoid colon specimens to 1 nM VCC resulted in an increase of I(SC by 20.7%, with respect to the basal values, while R(T was reduced by 12.3%. Moreover, increase in I(SC was abolished by bilateral Cl(- reduction. CONCLUSION/SIGNIFICANCE: Our results demonstrate that VCC, by forming anion channels on the apical membrane of enterocytes, triggers an outward transcellular flux of chloride. Such an ion movement, associated with the outward movement of Na(+ and water, might be responsible for the diarrhoea caused by the non-toxigenic strains of Vibrio cholerae.

  11. Comparison of classifications of aptamers against Vibrio ...

    African Journals Online (AJOL)

    As a novel method to detect the pathogen Vibrio alginolyticus, 45 aptamers were previously selected and tested. In order to better understand the properties of these aptamers, it was essential to classify these aptamers based on appropriate criteria. The primary structure of 45 aptamers against V. alginolyticus was analyzed ...

  12. Organic metabolites produced by Vibrio parahaemolyticus strain ...

    African Journals Online (AJOL)

    Identification and action of several antibacterial metabolites produced by a fish pathogen Vibrio parahaemolyticus strain An3 from marine ecosystem of Goa has been demonstrated. Antibacterial activity of the crude cell extract of the test bacterium has been evaluated against indicator pathogenic bacterial strains such as ...

  13. AKTIVITAS ANTIBAKTERI EKSTRAK BUAH ADAS (Foeniculum vulgare, Mill PADA Vibrio harveyi DAN Vibrio alginolyticus Antibacterial Activity of Fennel (Foeniculum vulgare Mill Extract on Vibrio alginolyticus and Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    Budianto Budianto

    2015-10-01

    Pada penelitian ini menggunakan ekstrak air dari buah adas untuk mengetahui aktivitas antibakteri terhadap Vibrio harveyi dan Vibrio alginolyticus dengan menggunakan metode uji Minimum Inhibitory Concentration (MIC dan difusi cakram kertas. Hasil yang diperoleh pada uji MIC, konsentrasi terkecil untuk menghambat pertumbuhan adalah 0,060 g/ml, untuk kedua spesies bakteri. Variasi perlakuan pada uji cakram kertas yaitu konsentrasi A (0,065 g/ml, B (0,070 g/ml, C (0,075 g/ml, D (0,080 g/ml, E (0,085 g/ml, F (0,090 g/ml dan kontrol (0,000 g/ml, hasil yang diperoleh adalah konsentrasi 0,090 g/ml memiliki diameter zona hambat tertinggi sebesar 11,17 ± 0,5 mm (V. harveyi dan 12,53 ± 1,14 mm (V. alginolyticus, sehingga dapat disimpulkan bahwa buah adas (F. vulgare Mill memiliki peranan ekologi yang sangat penting sebagai bahan pengobatan alternatif dalam pengendalian penyebaran penyakit Vibriosis yang disebabkan oleh V. harveyi dan V. alginolyticus. Kata kunci: Foeniculum vulgare Mill, Vibrio harveyi, Vibrio alginolyticus, uji MIC dan difusi cakram kertas

  14. Plankton composition and environmental factors contribute to Vibrio seasonality.

    Science.gov (United States)

    Turner, Jeffrey W; Good, Brooks; Cole, Dana; Lipp, Erin K

    2009-09-01

    Plankton represent a nutrient-rich reservoir capable of enriching Vibrio species, which can include human pathogens, at higher densities than the surrounding water column. To better understand the relationship between vibrios and plankton, the partitioning of culturable vibrios, on TCBS, between free living and plankton associated (63-200- and >200-microm-size fractions) was monitored over a 1-year period in coastal waters of Georgia, USA. Seasonal changes in the total Vibrio concentration were then compared with changes in environmental parameters as well as changes in the relative composition of the plankton community. Using univariate analyses, Vibrio concentrations were strongly associated with temperature, especially when those vibrios were plankton associated (R(2)=0.69 and 0.88 for the water and both plankton fractions; respectively) (Pplankton fractions were also correlated to shifts in the relative abundance of specific plankton taxa. In the 63-200-micro fraction, Vibrio concentrations were inversely associated with copepods, cyanobacteria and diatoms. In the >200-micro fraction, Vibrio concentrations were positively associated with copepods and negatively associated with decapod larvae. Our results confirm the role of temperature in Vibrio seasonality and highlight an important and independent role for plankton composition in explaining seasonal changes in Vibrio concentration.

  15. Identification of genes, desR and desA, required for utilization of desferrioxamine B as a xenosiderophore in Vibrio furnissii.

    Science.gov (United States)

    Tanabe, Tomotaka; Funahashi, Tatsuya; Miyamoto, Katsushiro; Tsujibo, Hiroshi; Yamamoto, Shigeo

    2011-01-01

    We found that Vibrio (V.) furnissii ATCC35016 can gain iron through a xenosiderophore desferrioxamine B (DFOB) for its growth under iron-limiting conditions, concurrent with the expression of the 79-kDa iron-repressible outer membrane protein (IROMP) in response to the presence of DFOB. Based on the sequence of the ferrioxamine B (an iron-bound form of DFOB) receptor gene in V. vulnificus, two V. furnissii genes, termed desA and desR, encoding the 79-kDa IROMP and AraC-type transcriptional regulator, respectively, were identified and cloned. Nucleotide sequences located in the promoter regions of both desR and desA predicted the presence of consensus ferric uptake regulation (Fur)-binding sequences. The transcription of both genes was negatively regulated by exogenous iron levels. Deletion of the desA gene abolished the ability of V. furnissii to utilize DFOB, and neither desA mRNA nor DesA was detected in the deletion mutant of desR regardless of the presence of DFOB. The functions of DesA and DesR as the ferrioxamine B receptor and transcriptional activator for desA, respectively, were confirmed by complementation of desA and desR deletion mutants.

  16. Prevalence of Salmonella and Vibrio spp. in seafood products sold in Singapore.

    Science.gov (United States)

    Huang, Yunle; Ghate, Vinayak; Phua, Leslie; Yuk, Hyun-Gyun

    2012-07-01

    Foodborne hazards in seafood have only recently received increased attention in Singapore since the illness outbreak in 2009 that was associated with consumption of Indian rojak (a traditional salad of fruits, vegetables, and seafood). The microbiological quality of seafood must be evaluated for assurance of food safety. The aim of this study was to evaluate the microbiological quality and to determine the prevalence of Salmonella and Vibrio spp. in seafood sold in Singapore. A total of 116 samples (41 prawn, 44 shellfish, and 31 fishball samples) were collected from major supermarkets and wet markets in Singapore. The mesophilic and psychrotrophic bacterial counts for prawn, shellfish, and fishballs were 2 to 7 log CFU/g. One Salmonella Lexington strain was isolated from a thawed-frozen shellfish product and two Vibrio parahaemolyticus strains were isolated from commercial fishball and shrimp meat products. Thus, seafood sold in Singapore has the potential to be contaminated with Vibrio spp. and Salmonella, and proper handling at food service establishments is required to ensure food safety. Effective control measures also are needed to prevent cross-contamination during postharvest seafood processing.

  17. A single regulatory gene is sufficient to alter Vibrio aestuarianus pathogenicity in oysters.

    Science.gov (United States)

    Goudenège, David; Travers, Marie Agnès; Lemire, Astrid; Petton, Bruno; Haffner, Philippe; Labreuche, Yannick; Tourbiez, Delphine; Mangenot, Sophie; Calteau, Alexandra; Mazel, Didier; Nicolas, Jean Louis; Jacq, Annick; Le roux, Frédérique

    2015-11-01

    Oyster diseases caused by pathogenic vibrios pose a major challenge to the sustainability of oyster farming. In France, since 2012 a disease affecting specifically adult oysters has been associated with the presence of Vibrio aestuarianus. Here, by combining genome comparison, phylogenetic analyses and high-throughput infections of strains isolated before or during the recent outbreaks, we show that virulent strains cluster into two V. aestuarianus lineages independently of the sampling dates. The bacterial lethal dose was not different between strains isolated before or after 2012. Hence, the emergence of a new highly virulent clonal strain is unlikely. Each lineage comprises nearly identical strains, the majority of them being virulent, suggesting that within these phylogenetically coherent virulent lineages a few strains have lost their pathogenicity. Comparative genomics allowed the identification of a single frameshift in a non-virulent strain. This mutation affects the varS gene that codes for a signal transduction histidine-protein kinase. Genetic analyses confirmed that varS is necessary for infection of oysters and for a secreted metalloprotease expression. For the first time in a Vibrio species, we show here that VarS is a key factor of pathogenicity. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

  18. [Prophage CTXphi genome variability and its role in alteration of Vibrio cholerae El Tor virulence characteristics].

    Science.gov (United States)

    Smirnova, N I; Osin, A V; Nefedov, K S; Kul'shan', T A; Zadnova, S P; Livanova, L F; Toporkov, A V; Kutyrev, V V

    2007-01-01

    Comparative analysis of CTXphi prophage genome of 366 V. cholerae El Tor strains isolated from infected people and water was carried out using the polymerase chain reaction. Four groups of vibrios, which carry different combinations of ctxA, zot, and ace genes from core region of CTXphi prophage coding key (cholera enterotoxin) and accessory (Zot and Ace toxins) pathogenicity factors, were determined: ctxA(+) zot(-) ace(+), ctxA(-) zot(+) ace(+), ctxA(-) zot(+) ace(-), ctxA(-) zot(-) ace(+). Vibrios that had lost all tested genes were also revealed. Genomic rearrangements occurring in water environment in virulent V. cholerae strains, which acquired foreign pathogenicity genes necessary for their existence in human organism, were proposed as one of the mechanisms of formation of clones with an incomplete or no prophage. Infection process in model animals challenged with wild and isogenic strains of V. cholerae differing in the set of the phage genes (ctxA, zot, and ace) was comparatively analyzed. It was shown that variability of CTXphi prophage genome was an important factor of modification of cholera vibrios virulent characteristics. Obtained data point to usefulness of ctxA, zot, and ace phage genes detection in wild V. cholerae isolates as it could permit evaluation of their virulent potential determining the severity of the infection.

  19. Vibrios in hatchery cultures of the razor clam, Solen marginatus (Pulteney).

    Science.gov (United States)

    Prado, S; Dubert, J; da Costa, F; Martínez-Patiño, D; Barja, J L

    2014-03-01

    Hatchery culture of the razor clam, Solen marginatus (Pulteney), has recently been developed in Galicia (NW Spain). However, recurrent episodes of mortalities of larval and post-larval cultures have been recorded during the course of various studies. The disease signs were similar to those described for other bivalve species in outbreaks caused by bacteria of the genus Vibrio. In this article, we present the results of microbiological monitoring of two batches of razor clams with different survival rates. All fermentative isolates were identified as members of the Splendidus clade within the genus Vibrio. Some of these isolates, identified as Vibrio splendidus-like, were clearly associated with the batch suffering mortalities, indicating their possible role as pathogens. Similar strains were found in the broodstock, suggesting vertical transmission of these bacteria. This is the first study of the microbiota associated with hatchery culture of S. marginatus, and the results will provide useful information for the optimization of a protocol for hatchery culture of this bivalve species. © 2013 John Wiley & Sons Ltd.

  20. Prevalence and population analysis of Vibrio parahaemolyticus in aquatic products from South China markets.

    Science.gov (United States)

    Xie, Tengfei; Wu, Qingping; Xu, Xiaoke; Zhang, Jumei; Guo, Weipeng

    2015-11-01

    Vibrio parahaemolyticus is a common foodborne pathogen in aquatic products. To investigate the prevalence of Vibrio parahaemolyticus in aquatic products in South China, 224 samples were collected from markets in four provinces (11 cities) from May 2013 to January 2014. One hundred and fifty isolates were isolated from 98 samples. All isolates were analyzed for the presence of thermostable direct haemolysin (TDH) and TDH-related haemolysin (TRH) by PCR, antibiotic susceptibility analysis by disk diffusion method, serotyping by multiplex PCR and molecular typing by enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) typing. Although all 150 isolates were negative for tdh, 61 strains were trh positive (40.67%). Antimicrobial susceptibility results indicated that most strains were resistant to streptomycin (88.67%), cefazolin (66.00%) and ampicillin (62.67%). All strains were susceptible to chloramphenicol. Forty percent of all isolates were O2 type. The 150 isolates were grouped into three clusters by ERIC-PCR typing. The results demonstrated the presence of V. parahaemolyticus in aquatic products from the retail market and this methodology can be used for microbiological risk assessment in China. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. [Detection of Salmonella, Listeria spp., Vibrio spp., and Yersinia enterocolitica in frozen seafood and comparison with enumeration for faecal indicators: implication for public health].

    Science.gov (United States)

    Ripabelli, G; Sammarco, M L; Fanelli, I; Grasso, G M

    2004-01-01

    Infections transmitted through consumption of contaminated seafood is a significant source of human morbidity. The aim of this study was to compare the detection of Salmonella, Listeria, Vibrio, and Yersinia enterocolitica in frozen seafood with results from enumeration of conventional faecal indicators. A total of 213 crustaceans or molluscs were purchased from local vendors in Italy: 74% were harvested in Italy, 25% from other European countries and 1% from outside Europe. Listeria spp. was isolated from 20% of samples, Vibrio spp. from 11%, Salmonella from 3% and Y. enterocolitica from 1%. Listeria species isolated were L. monocytogenes, L. innocua, L. welshimeri, L. ivanovii and L. seeligeri. Vibrio species isolated were V. alginolyticus and V. fluvialis. The most contaminated shellfish for both faecal indicator microrganism and pathogens were hen clams (6% contained Salmonella, 27% Listeria spp. and 3% Y. enterocolitica), while from 27% of shrimps Vibrio spp. was recovered. Higher levels of faecal indicators were recovered from samples harvested outside Europe, and 66% of samples harvested in Thailand were contaminated from Salmonella. Significant differences were found in the levels of contamination of seafoods depending upon the freezing regime, but there was a limited association between presence of potential pathogens (particularly Vibrio spp.) and conventional faecal indicators. Hence, we suggest reconsideration of current legal parameters to evaluate microbiological quality of seafood.

  2. Tropical Atlantic marine macroalgae with bioactivity against virulent and antibiotic resistant Vibrio

    Directory of Open Access Journals (Sweden)

    Giselle Cristina Silva

    2013-03-01

    Full Text Available The antibacterial activity of ethanol, methanol, hexane and acetone-based extracts of the macroalgae Padina gymnospora (PG, Hypnea musciformes (HM, Ulva fasciata (UF and Caulerpa prolifera (CP was investigated. The disk diffusion method was used to evaluate the algae antimicrobial effect against standard strains of Vibrio parahaemolyticus, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella enterica and five virulent antibiotic-resistant strains of V. brasiliensis, V. xuii and V. navarrensis (isolated from the hemolymph of Litopenaeus vannamei. Ethanol extracts of PG and HM inhibited all Vibrio strains. E. coli and P. aeruginosa were only susceptible to ethanol extracts of PG. Among the methanol extracts, only UF was bioactive, inhibiting V. navarrensis. The observed inhibitory effect of ethanol extracts of PG, HM and UF against virulent antibiotic-resistant bacteria suggests these macroalgal species constitute a potential source of bioactive compounds.

  3. Structural Characterization of the Extracellular Polysaccharide from Vibrio cholerae O1 El-Tor

    Science.gov (United States)

    Yildiz, Fitnat; Fong, Jiunn; Sadovskaya, Irina; Grard, Thierry; Vinogradov, Evgeny

    2014-01-01

    The ability to form biofilms is important for environmental survival, transmission, and infectivity of Vibrio cholerae, the causative agent of cholera in humans. To form biofilms, V. cholerae produces an extracellular matrix composed of proteins, nucleic acids and a glycoconjugate, termed Vibrio exopolysaccharide (VPS). Here, we present the data on isolation and characterization of the polysaccharide part of the VPS (VPS-PS), which has the following structure: where α-D-Glc is partially (∼20%) replaced with α-D-GlcNAc. α-GulNAcAGly is an amide between 2-acetamido-2-deoxy-α-guluronic acid and glycine. Apparently, the polysaccharide is bound to a yet unidentified component, which gives it high viscosity and completely suppresses any NMR signals belonging to the sugar chains of the VPS. The only reliable method to remove this component at present is a treatment of the whole glycoconjugate with concentrated hydrochloric acid. PMID:24520310

  4. Characterization of clinical Vibrio parahaemolyticus strains in Zhoushan, China, from 2013 to 2014.

    Directory of Open Access Journals (Sweden)

    Hongling Wang

    Full Text Available Vibrio parahaemolyticus is recognized as major cause of foodborne illness of global public health concern. This study collected 107 strains of V. parahaemolyticus during active surveillance of diarrheal diseases in hospitals in Zhoushan during 2013 to 2014 and investigated their serotypes, virulence genes (tdh, trh, and orf8, antimicrobial resistance, and genotypes. The dominant serotypes of the 107 clinical strains were O3:K6, O4:K8, and O4:KUT with 87.9% and 3.7% of the strains carrying the virulence genes tdh and trh, respectively. Molecular typing by pulsed-field gel electrophoresis indicated divergence among the clinical strains. Most isolates were sensitive to the common antimicrobial agents used against the Vibrio species except ampicillin. We conclude that continuous surveillance of V. parahaemolyticus in diarrhea patients is a public health priority and is useful for conducting risk assessment of foodborne illnesses caused by V. parahaemolyticus.

  5. Predation response of Vibrio fischeri biofilms to bacterivorus protists.

    Science.gov (United States)

    Chavez-Dozal, Alba; Gorman, Clayton; Erken, Martina; Steinberg, Peter D; McDougald, Diane; Nishiguchi, Michele K

    2013-01-01

    Vibrio fischeri proliferates in a sessile, stable community known as a biofilm, which is one alternative survival strategy of its life cycle. Although this survival strategy provides adequate protection from abiotic factors, marine biofilms are still susceptible to grazing by bacteria-consuming protozoa. Subsequently, grazing pressure can be controlled by certain defense mechanisms that confer higher biofilm antipredator fitness. In the present work, we hypothesized that V. fischeri exhibits an antipredator fitness behavior while forming biofilms. Different predators representing commonly found species in aquatic populations were examined, including the flagellates Rhynchomonas nasuta and Neobodo designis (early biofilm feeders) and the ciliate Tetrahymena pyriformis (late biofilm grazer). V. fischeri biofilms included isolates from both seawater and squid hosts (Euprymna and Sepiola species). Our results demonstrate inhibition of predation by biofilms, specifically, isolates from seawater. Additionally, antiprotozoan behavior was observed to be higher in late biofilms, particularly toward the ciliate T. pyriformis; however, inhibitory effects were found to be widespread among all isolates tested. These results provide an alternative explanation for the adaptive advantage and persistence of V. fischeri biofilms and provide an important contribution to the understanding of defensive mechanisms that exist in the out-of-host environment.

  6. Transformation Experiment Using Bioluminescence Genes of "Vibrio fischeri."

    Science.gov (United States)

    Slock, James

    1995-01-01

    Bioluminescence transformation experiments show students the excitement and power of recombinant DNA technology. This laboratory experiment utilizes two plasmids of "Vibrio fischeri" in a transformation experiment. (LZ)

  7. 21 CFR 866.3930 - Vibrio cholerae serological reagents.

    Science.gov (United States)

    2010-04-01

    ... cholera caused by the bacterium Vibrio cholerae and provides epidemiological information on cholera. Cholera is an acute infectious disease characterized by severe diarrhea with extreme fluid and electrolyte...

  8. Carriage of vibrio species by shrimps harvested from the coastal ...

    African Journals Online (AJOL)

    Objectives: To determine the prevalence of Vibrio spp in unprocessed shrimps and their susceptibility to antibiotics. Design: A prospective study of Vibrio spp associated with shrimps harvested from the coastal waters of South West Cameroon. Setting: A laboratory based study at the Department of Life Sciences, University ...

  9. Molecular uptake of chitooligosaccharides through chitoporin from the marine bacterium Vibrio harveyi.

    Directory of Open Access Journals (Sweden)

    Wipa Suginta

    Full Text Available BACKGROUND: Chitin is the most abundant biopolymer in marine ecosystems. However, there is no accumulation of chitin in the ocean-floor sediments, since marine bacteria Vibrios are mainly responsible for a rapid turnover of chitin biomaterials. The catabolic pathway of chitin by Vibrios is a multi-step process that involves chitin attachment and degradation, followed by chitooligosaccharide uptake across the bacterial membranes, and catabolism of the transport products to fructose-6-phosphate, acetate and NH(3. PRINCIPAL FINDINGS: This study reports the isolation of the gene corresponding to an outer membrane chitoporin from the genome of Vibrio harveyi. This porin, expressed in E. coli, (so called VhChiP was found to be a SDS-resistant, heat-sensitive trimer. Immunoblotting using anti-ChiP polyclonal antibody confirmed the expression of the recombinant ChiP, as well as endogenous expression of the native protein in the V. harveyi cells. The specific function of VhChiP was investigated using planar lipid membrane reconstitution technique. VhChiP nicely inserted into artificial membranes and formed stable, trimeric channels with average single conductance of 1.8±0.13 nS. Single channel recordings at microsecond-time resolution resolved translocation of chitooligosaccharides, with the greatest rate being observed for chitohexaose. Liposome swelling assays showed no permeation of other oligosaccharides, including maltose, sucrose, maltopentaose, maltohexaose and raffinose, indicating that VhChiP is a highly-specific channel for chitooligosaccharides. CONCLUSION/SIGNIFICANCE: We provide the first evidence that chitoporin from V. harveyi is a chitooligosaccharide specific channel. The results obtained from this study help to establish the fundamental role of VhChiP in the chitin catabolic cascade as the molecular gateway that Vibrios employ for chitooligosaccharide uptake for energy production.

  10. An acute gastroenteritis outbreak of Vibrio parahaemolyticus O4:K55 in Nursing College, Thailand.

    Science.gov (United States)

    Jatapai, A; Moungthong, B; Thunyaharn, S; Huttayananont, S; Rangsin, R

    2010-08-01

    A cluster of acute gastroenteritis among nursing students was noticed on 13th September 2005. Between 13th and 17th September 2005, a retrospective cohort study was then conducted to identify the most likely cause of gastroenteritis at a nursing college in Bangkok, Thailand. Self-administered questionnaires, interviews, environmental investigations, and rectal swabs from all participants were carried out. In the investigation, 98.9% female nursing students were investigated and had completed the questionnaire, 49.4% of the participants were diagnosed to have acute gastroenteritis. The predominant symptoms were watery diarrhoea (90.8%) and abdominal cramps (71.3%). Of 28.9% of rectal swab isolates were identified as Vibrio parahaemolyticus O4:K55 (40.4%), Salmonella spp. (19.2%), Vibrio cholerae non O1/non O139/non O141 (11.5%), Aeromonas trota (3.9%), Vibrio alginolyticus (1.9%) and other co-infections (23.1%). The tdh gene was identified from all V. parahaemolyticus using multiplex PCR. The implicated food risk factor for gastroenteritis was boiled egg (adjusted prevalence rate ratio; PR=1.9, 95% CI, 1.04-3.79). However the bitter melon soup was not significantly associated for gastroenteritis (adjusted PR=1.3, 95% CI, 0.98-1.82). The population attributable fraction analysis indicated that boiled eggs item was an implicated food risk for this outbreak (PAF=45.8%). Vibrio parahaemolyticus O4:K55 was identified as a major causative agent for gastroenteritis in which the contaminated boiled eggs was a vehicle in this outbreak. Cross-contamination control should be emphasized in food operation plans among institutes.

  11. Genomic epidemiology of Vibrio cholerae O1 associated with floods, Pakistan, 2010.

    Science.gov (United States)

    Shah, Muhammad Ali; Mutreja, Ankur; Thomson, Nicholas; Baker, Stephen; Parkhill, Julian; Dougan, Gordon; Bokhari, Habib; Wren, Brendan W

    2014-01-01

    In August 2010, Pakistan experienced major floods and a subsequent cholera epidemic. To clarify the population dynamics and transmission of Vibrio cholerae in Pakistan, we sequenced the genomes of all V. cholerae O1 El Tor isolates and compared the sequences to a global collection of 146 V. cholerae strains. Within the global phylogeny, all isolates from Pakistan formed 2 new subclades (PSC-1 and PSC-2), lying in the third transmission wave of the seventh-pandemic lineage that could be distinguished by signature deletions and their antimicrobial susceptibilities. Geographically, PSC-1 isolates originated from the coast, whereas PSC-2 isolates originated from inland areas flooded by the Indus River. Single-nucleotide polymorphism accumulation analysis correlated river flow direction with the spread of PSC-2. We found at least 2 sources of cholera in Pakistan during the 2010 epidemic and illustrate the value of a global genomic data bank in contextualizing cholera outbreaks.

  12. Monitoring water sources for environmental reservoirs of toxigenic Vibrio cholerae O1, Haiti.

    Science.gov (United States)

    Alam, Meer T; Weppelmann, Thomas A; Weber, Chad D; Johnson, Judith A; Rashid, Mohammad H; Birch, Catherine S; Brumback, Babette A; Beau de Rochars, Valery E Madsen; Morris, J Glenn; Ali, Afsar

    2014-03-01

    An epidemic of cholera infections was documented in Haiti for the first time in more than 100 years during October 2010. Cases have continued to occur, raising the question of whether the microorganism has established environmental reservoirs in Haiti. We monitored 14 environmental sites near the towns of Gressier and Leogane during April 2012-March 2013. Toxigenic Vibrio cholerae O1 El Tor biotype strains were isolated from 3 (1.7%) of 179 water samples; nontoxigenic O1 V. cholerae was isolated from an additional 3 samples. All samples containing V. cholerae O1 also contained non-O1 V. cholerae. V. cholerae O1 was isolated only when water temperatures were ≥31°C. Our data substantiate the presence of toxigenic V. cholerae O1 in the aquatic environment in Haiti. These isolations may reflect establishment of long-term environmental reservoirs in Haiti, which may complicate eradication of cholera from this coastal country.

  13. US Gulf-like toxigenic O1 Vibrio cholerae causing sporadic cholera outbreaks in China.

    Science.gov (United States)

    Luo, Yun; Octavia, Sophie; Jin, Dazhi; Ye, Julian; Miao, Ziping; Jiang, Tao; Xia, Shichang; Lan, Ruiting

    2016-05-01

    Cholera is potentially a life threatening disease caused by toxigenic Vibrio cholerae. Here we report the identification and characterisation of 76 non-7th pandemic clone O1 V. cholerae isolates including 65 clinical isolates from diarrhoeal patients from 2005 to 2014 in Zhejiang Province, China. We used multilocus sequence typing (MLST) to characterise 65 V. cholerae isolates. Pulse-Field Gel Electrophoresis (PFGE) was performed on a subset of the isolates and whole-genome sequencing was done on 13 isolates. MLST separated 65 isolates into 19 sequence types (STs). Thirty three isolates belonged to ST75 which also contains the US Gulf Coast clone. PFGE separated the 33 isolates into 16 pulsotypes. Whole genome sequencing of 10 ST75 isolates showed that the US Gulf Coast clone and the Chinese ST75 isolates can be separated into two distinct lineages, ST75a and ST75b. All Zhejiang ST75 isolates were ST75b. PFGE and genome sequencing confirmed the linked cases and identified small outbreaks caused by ST75b. The emergence and potential spread of ST75b may pose significant threat to public health. Epidemiological surveillance is required to further understand its epidemic potential. Copyright © 2016 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

  14. Vibrio fischeri metabolism: symbiosis and beyond.

    Science.gov (United States)

    Dunn, Anne K

    2012-01-01

    Vibrio fischeri is a bioluminescent, Gram-negative marine bacterium that can be found free living and in a mutualistic association with certain squids and fishes. Over the past decades, the study of V. fischeri has led to important discoveries about bioluminescence, quorum sensing, and the mechanisms that underlie beneficial host-microbe interactions. This chapter highlights what has been learned about metabolic pathways in V. fischeri, and how this information contributes to a broader understanding of the role of bacterial metabolism in host colonization by both beneficial and pathogenic bacteria, as well as in the growth and survival of free-living bacteria. Copyright © 2012 Elsevier Ltd. All rights reserved.

  15. The Vibrio parahaemolyticus Type III Secretion Systems manipulate host cell MAPK for critical steps in pathogenesis.

    LENUS (Irish Health Repository)

    Matlawska-Wasowska, Ksenia

    2010-12-01

    Vibrio parahaemolyticus is a food-borne pathogen causing inflammation of the gastrointestinal epithelium. Pathogenic strains of this bacterium possess two Type III Secretion Systems (TTSS) that deliver effector proteins into host cells. In order to better understand human host cell responses to V. parahaemolyticus, the modulation of Mitogen Activated Protein Kinase (MAPK) activation in epithelial cells by an O3:K6 clinical isolate, RIMD2210633, was investigated. The importance of MAPK activation for the ability of the bacterium to be cytotoxic and to induce secretion of Interleukin-8 (IL-8) was determined.

  16. Purification of Protease from Vibrio cholerae O1 and its Partial Characterization

    OpenAIRE

    Ichinose, Yoshio; Ehara, Masahiko; Utsunomiya, Akiyoshi

    1992-01-01

    A protease produced by a clinical isolate of Vibrio cholerae O1 was purified to apparent homogeneity by ammonium sulfate fractionation, ultracentrifuge and succesive column chromatography on a Bio-gel A5m and a TSK gel G-3000SW HPLC column. The molecular weight of the purified protease was estimated to be 32,000 on the basis of its mobility on SDS-PAGE and was identical to HA/protease biochemically and physicochemically as previously reported (Booth et al., 1983; Honda et al., 1987). The hema...

  17. Fuse or die: how to survive the loss of Dam in Vibrio cholerae

    DEFF Research Database (Denmark)

    Val, Marie-Eve; Kennedy, Sean P; Soler-Bistue, Alfonso J.

    2014-01-01

    Dam methylates GATC sequences in γ-proteobacteria genomes, regulating several cellular functions including replication. In Vibrio cholerae, which has two chromosomes, Dam is essential for viability, owing to its role in chr2 replication initiation. In this study, we isolated spontaneous mutants...... of V. cholerae that were able to survive the deletion of dam. In these mutants, homologous recombination and chromosome dimer resolution are essential, unless DNA mismatch repair is inactivated. Furthermore, the initiator of chr2 replication, RctB, is no longer required. We show that, instead...

  18. Antibiotic Resistance and Integron of Vibrio cholerae Detection from School Street Foods in Jakarta

    OpenAIRE

    DEASHINTA, NADIA; WATURANGI, DIANA ELIZABETH; YOGIARA,

    2007-01-01

    Street foods represent foods and beverages prepared by vendors in streets or other public places, i.e. schools. Food safety issues perceive street foods as a potential major public risk. Street foods contaminated with toxigenic Vibrio cholerae may lead to serious poisoning to school-age children. In this study, 17 isolates of V. cholerae were obtained from nine (45%) of total 20 street foods samples collected in Jakarta. Five (29%) were confirmed to be V. cholerae O1, serotype Ogawa using bio...

  19. Evolutionary dynamics of Vibrio cholerae O1 following a single-source introduction to Haiti.

    Science.gov (United States)

    Katz, Lee S; Petkau, Aaron; Beaulaurier, John; Tyler, Shaun; Antonova, Elena S; Turnsek, Maryann A; Guo, Yan; Wang, Susana; Paxinos, Ellen E; Orata, Fabini; Gladney, Lori M; Stroika, Steven; Folster, Jason P; Rowe, Lori; Freeman, Molly M; Knox, Natalie; Frace, Mike; Boncy, Jacques; Graham, Morag; Hammer, Brian K; Boucher, Yan; Bashir, Ali; Hanage, William P; Van Domselaar, Gary; Tarr, Cheryl L

    2013-07-02

    Prior to the epidemic that emerged in Haiti in October of 2010, cholera had not been documented in this country. After its introduction, a strain of Vibrio cholerae O1 spread rapidly throughout Haiti, where it caused over 600,000 cases of disease and >7,500 deaths in the first two years of the epidemic. We applied whole-genome sequencing to a temporal series of V. cholerae isolates from Haiti to gain insight into the mode and tempo of evolution in this isolated population of V. cholerae O1. Phylogenetic and Bayesian analyses supported the hypothesis that all isolates in the sample set diverged from a common ancestor within a time frame that is consistent with epidemiological observations. A pangenome analysis showed nearly homogeneous genomic content, with no evidence of gene acquisition among Haiti isolates. Nine nearly closed genomes assembled from continuous-long-read data showed evidence of genome rearrangements and supported the observation of no gene acquisition among isolates. Thus, intrinsic mutational processes can account for virtually all of the observed genetic polymorphism, with no demonstrable contribution from horizontal gene transfer (HGT). Consistent with this, the 12 Haiti isolates tested by laboratory HGT assays were severely impaired for transformation, although unlike previously characterized noncompetent V. cholerae isolates, each expressed hapR and possessed a functional quorum-sensing system. Continued monitoring of V. cholerae in Haiti will illuminate the processes influencing the origin and fate of genome variants, which will facilitate interpretation of genetic variation in future epidemics. Vibrio cholerae is the cause of substantial morbidity and mortality worldwide, with over three million cases of disease each year. An understanding of the mode and rate of evolutionary change is critical for proper interpretation of genome sequence data and attribution of outbreak sources. The Haiti epidemic provides an unprecedented opportunity to

  20. The Role of Vibrios in Diseases of Corals.

    Science.gov (United States)

    Munn, Colin B

    2015-08-01

    The tissue, skeleton, and secreted mucus of corals supports a highly dynamic and diverse community of microbes, which play a major role in the health status of corals such as the provision of essential nutrients or the metabolism of waste products. However, members of the Vibrio genus are prominent as causative agents of disease in corals. The aim of this chapter is to review our understanding of the spectrum of disease effects displayed by coral-associated vibrios, with a particular emphasis on the few species where detailed studies of pathogenicity have been conducted. The role of Vibrio shilonii in seasonal bleaching of Oculina patagonica and the development of the coral probiotic hypothesis is reviewed, pointing to unanswered questions about this phenomenon. Detailed consideration is given to studies of V. coralliilyticus and related pathogens and changes in the dominance of vibrios associated with coral bleaching. Other Vibrio-associated disease syndromes discussed include yellow band/blotch disease and tissue necrosis in temperate gorgonian corals. The review includes analysis of the role of enzymes, resistance to oxidative stress, and quorum sensing in virulence of coral-associated vibrios. The review concludes that we should probably regard most-possibly all-vibrios as "opportunistic" pathogens which, under certain environmental conditions, are capable of overwhelming the defense mechanisms of appropriate hosts, leading to rapid growth and tissue destruction.

  1. Ocurrence of Vibrio spp., positive coagulase staphylococci and enteric bacteria in oysters (Crassostrea gigas harvested in the south bay of Santa Catarina island, Brazil Ocorrência de Vibrio spp., estafilococos coagulase positivo e bactérias entéricas em ostras (Crassostrea gigas cultivadas na baía sul da ilha de Santa Catarina, Brasil

    Directory of Open Access Journals (Sweden)

    Roberta Juliano Ramos

    2012-09-01

    Full Text Available The aim of this study was to assess the contamination of oysters (Crassostrea gigas, harvested in six different regions of the South Bay of Santa Catarina Island, with Coliforms at 45 ºC, Escherichia coli, Vibrio spp., positive coagulase staphylococci, and Salmonella sp. over a period of one year. One hundred eighty oyster samples were collected directly from their culture sites and analyzed. Each sample consisted of a pool of 12 oysters. All of the samples analyzed showed absence of Salmonella, 18 (10% samples showed presence of Escherichia coli, 15 (8.3% samples were positive for V. alginolyticus, and Vibriocholerae was detected in 4 samples (2.2%. The counts of positive-coagulase staphylococci varied from O objetivo deste estudo foi avaliar a contaminação de ostras (Crassostrea gigas cultivadas em diferentes regiões da Baía Sul da Ilha de Santa Catarina, por coliformes a 45 ºC, Escherichia coli, Vibrio spp. Estafilococos coagulase positiva e Salmonella sp., durante o período de um ano. Foram analisadas 180 amostras, coletadas diretamente no local de cultivo. Todas as amostras analisadas apresentaram ausência de Salmonella, 18 (10% amostras apresentaram presença de Escherichia coli, 15 (8,3% amostras positivas para Vibrio alginolyticus e V. cholerae foi detectado em 4 amostras (2,2%. As contagens de Estafilococos coagulase positiva variaram de <10 a 1,9 x 102 UFC.g-1, enquanto que as contagens de coliformes a 45 ºC e E. coli variaram de <3 a 1,5 x 102 NMP.g-1 e <3 e 4,3 x 10 NMP.g-1, respectivamente. As contagens de V. parahaemolyticus e V. vulnificus variaram de <3 a 7 NMP.g-1, para ambos os microrganismos, sugerindo um monitoramento tanto destas espécies quanto da temperatura das águas marinhas nas regiões de cultivo. Com base nos resultados das análises microbiológicas, as amostras analisadas mostraram qualidade bacteriológica aceitável, ou seja, dentro dos parâmetros estabelecidos na legislação brasileira.

  2. Environmental reservoirs and mechanisms of persistence of Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Carla eLutz

    2013-12-01

    Full Text Available It is now well accepted that Vibrio cholerae, the causative agent of the water-borne disease cholera, is acquired from environmental sources where it persists between outbreaks of the disease. Recent advances in molecular technology have demonstrated that this bacterium could be detected in areas where it had not been isolated from before, indicating a much broader, global distribution of this bacterium rather than specifically within regions where cholera is endemic. The environmental persistence of V. cholerae in the aquatic environment can be attributed to multiple intra- and interspecific strategies such as responsive gene regulation and biofilm formation on biotic and abiotic surfaces, as well as interactions with a multitude of other organisms. This review will discuss some of the mechanisms that enable the persistence of the bacterium in the sometimes hostile environment. In particular, we will discuss how V. cholerae can survive stressors such as starvation, temperature and salinity fluctuations as well as how the organism persists under constant predation by heterotrophic protists.

  3. Vibrio cholerae: A historical perspective and current trend

    Directory of Open Access Journals (Sweden)

    Mary Oyenike Oladokun

    2016-11-01

    Full Text Available Vibrio cholerae (V. cholerae is a Gram-negative, curved, rod-shaped bacteria with two of its strains V. cholerae O1 and V. cholerae O139 known to cause cholera, a deadly diarrheal disease that has repeatedly plagued the world in pandemics since 1817 and still remains a public health problem globally till today. The pathogens’ persistence in aquatic milieux during inter-epidemic periods is facilitated by the production of a biofilm, thus evolving from being an infection of oral-fecal transmission to a more composite ecological framework of a communicable disease. The outbreaks of cholera spread rapidly in various intensities within and among countries and even continents and the World Health Organization estimates that 3–5 million cases outbreak and over 200 000 die yearly from cholera. Also, the impact of a cholera epidemic is not limited to its high morbidity and mortality rates alone, but also the grievous impact on the economy of the countries experiencing the outbreaks. In this review, we carried out an overview of V. cholerae including its isolation and detection, genetics as well as a comparison of the toxigenic and non-toxigenic determinants in the human host and the host defences. Furthermore, the history of global pandemics, cost implications, conflict and ecological methodologies of cholera prevention and control. The management of disease and antibiotic resistance in V. cholerae are also highlighted.

  4. Directed evolution of Vibrio fischeri LuxR signal sensitivity.

    Science.gov (United States)

    Kimura, Yuki; Tashiro, Yohei; Saito, Kyoichi; Kawai-Noma, Shigeko; Umeno, Daisuke

    2016-11-01

    LuxR is the core component of Vibrio fischeri quorum sensing. It acts as the transcriptional activator by binding to its cognate signaling molecules 3-oxo-hexanoyl-homoserine lactone (3OC6HSL). Although several acyl-HSLs with 3-oxo groups are known to activate LuxR with similar efficiency, acyl-HSLs without 3-oxo groups are very weak inducers. We conducted a round of LuxR directed evolution to acquire LuxR mutants with higher signal sensitivity to octanoyl-homoserine lactone (C8HSL). All of the isolated mutants showed increased signal sensitivity to many other acyl-HSLs, including C8HSL, and some to the LuxR antagonist p-coumaroyl-HSL. The evolution of their ligand sensitivity proceeded through the stabilization of the signal-bound state, thereby elevating the effective concentration of LuxR at the ON-state. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  5. The repertoire of glycosphingolipids recognized by Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    John Benktander

    Full Text Available The binding of cholera toxin to the ganglioside GM1 as the initial step in the process leading to diarrhea is nowadays textbook knowledge. In contrast, the knowledge about the mechanisms for attachment of Vibrio cholerae bacterial cells to the intestinal epithelium is limited. In order to clarify this issue, a large number of glycosphingolipid mixtures were screened for binding of El Tor V. cholerae. Several specific interactions with minor complex non-acid glycosphingolipids were thereby detected. After isolation of binding-active glycosphingolipids, characterization by mass spectrometry and proton NMR, and comparative binding studies, three distinct glycosphingolipid binding patterns were defined. Firstly, V. cholerae bound to complex lacto/neolacto glycosphingolipids with the GlcNAcβ3Galβ4GlcNAc sequence as the minimal binding epitope. Secondly, glycosphingolipids with a terminal Galα3Galα3Gal moiety were recognized, and the third specificity was the binding to lactosylceramide and related compounds. V. cholerae binding to lacto/neolacto glycosphingolipids, and to the other classes of binding-active compounds, remained after deletion of the chitin binding protein GbpA. Thus, the binding of V. cholerae to chitin and to lacto/neolacto containing glycosphingolipids represents two separate binding specificities.

  6. Antibiotic Resistance and Integron of Vibrio cholerae Detection from School Street Foods in Jakarta

    Directory of Open Access Journals (Sweden)

    NADIA DEASHINTA

    2007-06-01

    Full Text Available Street foods represent foods and beverages prepared by vendors in streets or other public places, i.e. schools. Food safety issues perceive street foods as a potential major public risk. Street foods contaminated with toxigenic Vibrio cholerae may lead to serious poisoning to school-age children. In this study, 17 isolates of V. cholerae were obtained from nine (45% of total 20 street foods samples collected in Jakarta. Five (29% were confirmed to be V. cholerae O1, serotype Ogawa using biochemical tests and serological identification. Of the 17 V. cholerae isolates 47% proved to be resistant to ampicillin, 35% to trimethoprim, 17.6% to tetracycline, and 17.6% to streptomycin. A class 1 integrons bearing streptomycin/spectinomycin resistant gene cassette of aadA1c were discovered on isolate Vc25n. This may leads to horizontal transfer of the antibiotic resistant genes to other bacteria.

  7. Use of REP- and ERIC-PCR to reveal genetic heterogeneity of Vibrio cholerae from edible ice in Jakarta, Indonesia

    Directory of Open Access Journals (Sweden)

    Waturangi Diana E

    2012-03-01

    Full Text Available Abstract Background Vibrio cholerae is the causative organism of waterborne disease, cholera. V. cholerae has caused many epidemics and pandemics of cholera for many years. In this study, V. cholerae recovered from edible ice were investigated for their genetic diversity using Enterobacterial Repetitive Intergenic Consensus (ERIC PCR and Repetitive Extragenic Palindromic (REP PCR. Isolation was done using selective medium and the presumptive isolates were confirmed through biochemical and serological assays. Results Seventy-five isolates of V. cholerae were recovered from ice samples collected from different locations of Jakarta. Specifically, 19 of them were identified as O1 serotype, 16 were Ogawa, 3 isolates were Inaba and the remaining isolates were non-O1. The fingerprinting profiles of V.cholerae isolated from ice samples were very diverse. Conclusion This result showed that the ERIC sequence is more informative and discriminative than REP sequence for analysis of V. cholerae diversity.

  8. Catechol Siderophore Transport by Vibrio cholerae.

    Science.gov (United States)

    Wyckoff, Elizabeth E; Allred, Benjamin E; Raymond, Kenneth N; Payne, Shelley M

    2015-09-01

    Siderophores, small iron-binding molecules secreted by many microbial species, capture environmental iron for transport back into the cell. Vibrio cholerae synthesizes and uses the catechol siderophore vibriobactin and also uses siderophores secreted by other species, including enterobactin produced by Escherichia coli. E. coli secretes both canonical cyclic enterobactin and linear enterobactin derivatives likely derived from its cleavage by the enterobactin esterase Fes. We show here that V. cholerae does not use cyclic enterobactin but instead uses its linear derivatives. V. cholerae lacked both a receptor for efficient transport of cyclic enterobactin and enterobactin esterase to promote removal of iron from the ferrisiderophore complex. To further characterize the transport of catechol siderophores, we show that the linear enterobactin derivatives were transported into V. cholerae by either of the catechol siderophore receptors IrgA and VctA, which also transported the synthetic siderophore MECAM [1,3,5-N,N',N″-tris-(2,3-dihydroxybenzoyl)-triaminomethylbenzene]. Vibriobactin is transported via the additional catechol siderophore receptor ViuA, while the Vibrio fluvialis siderophore fluvibactin was transported by all three catechol receptors. ViuB, a putative V. cholerae siderophore-interacting protein (SIP), functionally substituted for the E. coli ferric reductase YqjH, which promotes the release of iron from the siderophore in the bacterial cytoplasm. In V. cholerae, ViuB was required for the use of vibriobactin but was not required for the use of MECAM, fluvibactin, ferrichrome, or the linear derivatives of enterobactin. This suggests the presence of another protein in V. cholerae capable of promoting the release of iron from these siderophores. Vibrio cholerae is a major human pathogen and also serves as a model for the Vibrionaceae, which include other serious human and fish pathogens. The ability of these species to persist and acquire essential

  9. Opkomst van Vibrio infecties in brakwaterkweekvis : uit de ziekenboeg

    NARCIS (Netherlands)

    Haenen, O.L.M.; Engelsma, M.Y.

    2010-01-01

    De laatste paar jaar zijn er diverse Vibrio-soorten als ziekteverwekkende bacterie aangetoond in brak- en zoutwaterkweekvis. We gaan in dit artikel in op vibriose bij tong, tarbot, barramundi een zeebaars.

  10. Vibrio cholerae classical biotype strains reveal distinct signatures in Mexico.

    Science.gov (United States)

    Alam, Munirul; Islam, M Tarequl; Rashed, Shah Manzur; Johura, Fatema-tuz; Bhuiyan, Nurul A; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Watanabe, Haruo; Hasan, Nur-A; Colwell, Rita R; Cravioto, Alejandro

    2012-07-01

    Vibrio cholerae O1 classical (CL) biotype caused the fifth and sixth pandemics, and probably the earlier cholera pandemics, before the El Tor (ET) biotype initiated the seventh pandemic in Asia in the 1970s by completely displacing the CL biotype. Although the CL biotype was thought to be extinct in Asia and although it had never been reported from Latin America, V. cholerae CL and ET biotypes, including a hybrid ET, were found associated with areas of cholera endemicity in Mexico between 1991 and 1997. In this study, CL biotype strains isolated from areas of cholera endemicity in Mexico between 1983 and 1997 were characterized in terms of major phenotypic and genetic traits and compared with CL biotype strains isolated in Bangladesh between 1962 and 1989. According to sero- and biotyping data, all V. cholerae strains tested had the major phenotypic and genotypic characteristics specific for the CL biotype. Antibiograms revealed the majority of the Bangladeshi strains to be resistant to trimethoprim-sulfamethoxazole, furazolidone, ampicillin, and gentamicin, while the Mexican strains were sensitive to all of these drugs, as well as to ciprofloxacin, erythromycin, and tetracycline. Pulsed-field gel electrophoresis (PFGE) of NotI-digested genomic DNA revealed characteristic banding patterns for all of the CL biotype strains although the Mexican strains differed from the Bangladeshi strains in 1 to 2 DNA bands. The difference was subtle but consistent, as confirmed by the subclustering patterns in the PFGE-based dendrogram, and can serve as a regional signature, suggesting the pre-1991 existence and evolution of the CL biotype strains in the Americas, independent from Asia.

  11. Inactivation of Vibrio anguillarum by attached and planktonic Roseobacter cells

    DEFF Research Database (Denmark)

    D'Alvise, Paul; Melchiorsen, Jette; Porsby, Cisse Hedegaard

    2010-01-01

    The purpose of the present study was to investigate inhibition of Vibrio by Roseobacter in a combined liquid-surface system. Exposure of Vibrio anguillarum to surface-attached roseobacters (10e7 cfu/cm2) resulted in significant reduction or complete killing of the pathogen inoculated at 10e2 – 10...... cfu/ml. The effect was likely associated with production of tropodithietic acid (TDA), as a TDA-negative mutant did not affect survival or growth of V. anguillarum....

  12. Engineering Vibrio fischeri for Inducible Gene Expression.

    Science.gov (United States)

    Ondrey, Jakob M; Visick, Karen L

    2014-01-01

    The marine bacterium Vibrio fischeri serves as a model organism for a variety of natural phenomena, including symbiotic host colonization. The ease with which the V. fischeri genome can be manipulated contributes greatly to our ability to identify the factors involved in these phenomena. Here, we have adapted genetic tools for use in V. fischeri to promote our ability to conditionally control the expression of genes of interest. Specifically, we modified the commonly used mini-Tn5 transposon to contain an outward-facing, LacI-repressible/IPTG-inducible promoter, and inserted the lacI gene into the V. fischeri chromosome. Used together, these tools permit the identification and induction of genes that control specific phenotypes. To validate this approach, we identified IPTG-controllable motility mutants. We anticipate that the ability to randomly insert an inducible promoter into the genome of V. fischeri will advance our understanding of various aspects of the physiology of this microbe.

  13. Vibrio cholerae Biofilms and Cholera Pathogenesis.

    Directory of Open Access Journals (Sweden)

    Anisia J Silva

    2016-02-01

    Full Text Available Vibrio cholerae can switch between motile and biofilm lifestyles. The last decades have been marked by a remarkable increase in our knowledge of the structure, regulation, and function of biofilms formed under laboratory conditions. Evidence has grown suggesting that V. cholerae can form biofilm-like aggregates during infection that could play a critical role in pathogenesis and disease transmission. However, the structure and regulation of biofilms formed during infection, as well as their role in intestinal colonization and virulence, remains poorly understood. Here, we review (i the evidence for biofilm formation during infection, (ii the coordinate regulation of biofilm and virulence gene expression, and (iii the host signals that favor V. cholerae transitions between alternative lifestyles during intestinal colonization, and (iv we discuss a model for the role of V. cholerae biofilms in pathogenicity.

  14. Viscosity dictates metabolic activity of Vibrio ruber

    Science.gov (United States)

    Borić, Maja; Danevčič, Tjaša; Stopar, David

    2012-01-01

    Little is known about metabolic activity of bacteria, when viscosity of their environment changes. In this work, bacterial metabolic activity in media with viscosity ranging from 0.8 to 29.4 mPas was studied. Viscosities up to 2.4 mPas did not affect metabolic activity of Vibrio ruber. On the other hand, at 29.4 mPas respiration rate and total dehydrogenase activity increased 8 and 4-fold, respectively. The activity of glucose-6-phosphate dehydrogenase (GPD) increased up to 13-fold at higher viscosities. However, intensified metabolic activity did not result in faster growth rate. Increased viscosity delayed the onset as well as the duration of biosynthesis of prodigiosin. As an adaptation to viscous environment V. ruber increased metabolic flux through the pentose phosphate pathway and reduced synthesis of a secondary metabolite. In addition, V. ruber was able to modify the viscosity of its environment. PMID:22826705

  15. The population structure of Vibrio cholerae from the Chandigarh Region of Northern India.

    Directory of Open Access Journals (Sweden)

    Moataz Abd El Ghany

    2014-07-01

    Full Text Available Cholera infection continues to be a threat to global public health. The current cholera pandemic associated with Vibrio cholerae El Tor has now been ongoing for over half a century.Thirty-eight V. cholerae El Tor isolates associated with a cholera outbreak in 2009 from the Chandigarh region of India were characterised by a combination of microbiology, molecular typing and whole-genome sequencing. The genomic analysis indicated that two clones of V. cholera circulated in the region and caused disease during this time. These clones fell into two distinct sub-clades that map independently onto wave 3 of the phylogenetic tree of seventh pandemic V. cholerae El Tor. Sequence analyses of the cholera toxin gene, the Vibrio seventh Pandemic Island II (VSPII and SXT element correlated with this phylogenetic position of the two clades on the El Tor tree. The clade 2 isolates, characterized by a drug-resistant profile and the expression of a distinct cholera toxin, are closely related to the recent V. cholerae isolated elsewhere, including Haiti, but fell on a distinct branch of the tree, showing they were independent outbreaks. Multi-Locus Sequence Typing (MLST distinguishes two sequence types among the 38 isolates, that did not correspond to the clades defined by whole-genome sequencing. Multi-Locus Variable-length tandem-nucleotide repeat Analysis (MLVA identified 16 distinct clusters.The use of whole-genome sequencing enabled the identification of two clones of V. cholerae that circulated during the 2009 Chandigarh outbreak. These clones harboured a similar structure of ICEVchHai1 but differed mainly in the structure of CTX phage and VSPII. The limited capacity of MLST and MLVA to discriminate between the clones that circulated in the 2009 Chandigarh outbreak highlights the value of whole-genome sequencing as a route to the identification of further genetic markers to subtype V. cholerae isolates.

  16. The Population Structure of Vibrio cholerae from the Chandigarh Region of Northern India

    KAUST Repository

    Abd El Ghany, Moataz

    2014-07-24

    Background:Cholera infection continues to be a threat to global public health. The current cholera pandemic associated with Vibrio cholerae El Tor has now been ongoing for over half a century.Methodology/Principal Findings:Thirty-eight V. cholerae El Tor isolates associated with a cholera outbreak in 2009 from the Chandigarh region of India were characterised by a combination of microbiology, molecular typing and whole-genome sequencing. The genomic analysis indicated that two clones of V. cholera circulated in the region and caused disease during this time. These clones fell into two distinct sub-clades that map independently onto wave 3 of the phylogenetic tree of seventh pandemic V. cholerae El Tor. Sequence analyses of the cholera toxin gene, the Vibrio seventh Pandemic Island II (VSPII) and SXT element correlated with this phylogenetic position of the two clades on the El Tor tree. The clade 2 isolates, characterized by a drug-resistant profile and the expression of a distinct cholera toxin, are closely related to the recent V. cholerae isolated elsewhere, including Haiti, but fell on a distinct branch of the tree, showing they were independent outbreaks. Multi-Locus Sequence Typing (MLST) distinguishes two sequence types among the 38 isolates, that did not correspond to the clades defined by whole-genome sequencing. Multi-Locus Variable-length tandem-nucleotide repeat Analysis (MLVA) identified 16 distinct clusters.Conclusions/Significance:The use of whole-genome sequencing enabled the identification of two clones of V. cholerae that circulated during the 2009 Chandigarh outbreak. These clones harboured a similar structure of ICEVchHai1 but differed mainly in the structure of CTX phage and VSPII. The limited capacity of MLST and MLVA to discriminate between the clones that circulated in the 2009 Chandigarh outbreak highlights the value of whole-genome sequencing as a route to the identification of further genetic markers to subtype V. cholerae isolates.

  17. Multi-locus variable number tandem repeat analysis of 7th pandemic Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Lam Connie

    2012-05-01

    Full Text Available Abstract Background Seven pandemics of cholera have been recorded since 1817, with the current and ongoing pandemic affecting almost every continent. Cholera remains endemic in developing countries and is still a significant public health issue. In this study we use multilocus variable number of tandem repeats (VNTRs analysis (MLVA to discriminate between isolates of the 7th pandemic clone of Vibrio cholerae. Results MLVA of six VNTRs selected from previously published data distinguished 66 V. cholerae isolates collected between 1961–1999 into 60 unique MLVA profiles. Only 4 MLVA profiles consisted of more than 2 isolates. The discriminatory power was 0.995. Phylogenetic analysis showed that, except for the closely related profiles, the relationships derived from MLVA profiles were in conflict with that inferred from Single Nucleotide Polymorphism (SNP typing. The six SNP groups share consensus VNTR patterns and two SNP groups contained isolates which differed by only one VNTR locus. Conclusions MLVA is highly discriminatory in differentiating 7th pandemic V. cholerae isolates and MLVA data was most useful in resolving the genetic relationships among isolates within groups previously defined by SNPs. Thus MLVA is best used in conjunction with SNP typing in order to best determine the evolutionary relationships among the 7th pandemic V. cholerae isolates and for longer term epidemiological typing.

  18. Thiocarbamates from Moringa oleifera Seeds Bioactive against Virulent and Multidrug-Resistant Vibrio Species

    Science.gov (United States)

    de Sousa, Oscarina Viana; Hofer, Ernesto; Mafezoli, Jair; Barbosa, Francisco Geraldo

    2017-01-01

    Prospect of antibacterial agents may provide an alternative therapy for diseases caused by multidrug-resistant bacteria. This study aimed to evaluate the in vitro bioactivity of Moringa oleifera seed extracts against 100 vibrios isolated from the marine shrimp Litopenaeus vannamei. Ethanol extracts at low (MOS-E) and hot (MOS-ES) temperature are shown to be bioactive against 92% and 90% of the strains, respectively. The most efficient Minimum Inhibitory Concentration (MIC) levels of MOS-E and MOS-ES against a high percentage of strains were 32 µg mL−1. Bioguided screening of bioactive compounds showed that the ethyl acetate fraction from both extracts was the only one that showed antibacterial activity. Vibriocidal substances, niazirine and niazimicine, were isolated from the aforementioned fraction through chromatographic fractionation. PMID:28770224

  19. Roles of thermostable direct hemolysin (TDH and TDH-related hemolysin (TRH in Vibrio parahaemolyticus

    Directory of Open Access Journals (Sweden)

    Pendru eRaghunath

    2015-01-01

    Full Text Available Vibrio parahaemolyticus is the leading cause of seafood borne bacterial gastroenteritis in the world, often associated with the consumption of raw or undercooked seafood. However, not all strains of V. parahaemolyticus are pathogenic. The thermostable direct haemolysin (TDH or TDH-related haemolysin (TRH encoded by tdh and trh genes, respectively are considered major virulence factors in V. parahaemolyticus. However, about 10% of clinical strains do not contain tdh and/or trh. Environmental isolates of V. parahaemolyticus lacking tdh and/or trh are also highly cytotoxic to human gastrointestinal cells. Even in the absence of these hemolysins, V. parahaemolyticus remains pathogenic indicating other virulence factors exist. This mini review aims at discussing the possible roles of tdh and trh genes in clinical and environmental isolates of V. parahaemolyticus.

  20. Roles of thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) in Vibrio parahaemolyticus.

    Science.gov (United States)

    Raghunath, Pendru

    2014-01-01

    Vibrio parahaemolyticus is the leading cause of seafood borne bacterial gastroenteritis in the world, often associated with the consumption of raw or undercooked seafood. However, not all strains of V. parahaemolyticus are pathogenic. The thermostable direct hemolysin (TDH) or TDH-related hemolysin (TRH) encoded by tdh and trh genes, respectively, are considered major virulence factors in V. parahaemolyticus. However, about 10% of clinical strains do not contain tdh and/or trh. Environmental isolates of V. parahaemolyticus lacking tdh and/or trh are also highly cytotoxic to human gastrointestinal cells. Even in the absence of these hemolysins, V. parahaemolyticus remains pathogenic indicating other virulence factors exist. This mini review aims at discussing the possible roles of tdh and trh genes in clinical and environmental isolates of V. parahaemolyticus.

  1. Genome-wide characterization of vibrio phage ϕpp2 with unique arrangements of the mob-like genes

    Directory of Open Access Journals (Sweden)

    Lin Ying-Rong

    2012-06-01

    Full Text Available Abstract Background Vibrio parahaemolyticus is associated with gastroenteritis, wound infections, and septicemia in human and animals. Phages can control the population of the pathogen. So far, the only one reported genome among giant vibriophages is KVP40: 244,835 bp with 26% coding regions that have T4 homologs. Putative homing endonucleases (HE were found in Vibrio phage KVP40 bearing one segD and Vibrio cholerae phage ICP1 carrying one mobC/E and one segG. Results A newly isolated Vibrio phage ϕpp2, which was specific to the hosts of V. parahaemolyticus and V. alginolyticus, featured a long nonenveloped head of ~90 × 150 nm and tail of ~110 nm. The phage can survive at 50°C for more than one hour. The genome of the phage ϕpp2 was sequenced to be 246,421 bp, which is 1587 bp larger than KVP40. 383 protein-encoding genes (PEGs and 30 tRNAs were found in the phage ϕpp2. Between the genomes of ϕpp2 and KVP40, 254 genes including 29 PEGs for viral structure were of high similarity, whereas 17 PEGs of KVP40 and 21 PEGs of ϕpp2 were unmatched. In both genomes, the capsid and tail genes have been identified, as well as the extensive representation of the DNA replication, recombination, and repair enzymes. In addition to the three giant indels of 1098, 1143 and 3330 nt, ϕpp2 possessed unique proteins involved in potassium channel, gp2 (DNA end protector, tRNA nucleotidyltransferase, and mob-type HEs, which were not reported in KVP40. The ϕpp2 PEG274, with strong promoters and translational initiation, was identified to be a mobE type, flanked by NrdA and NrdB/C homologs. Coincidently, several pairs of HE-flanking homologs with empty center were found in the phages of Vibrio phages ϕpp2 and KVP40, as well as in Aeromonas phages (Aeh1 and Ae65, and cyanophage P-SSM2. Conclusions Vibrio phage ϕpp2 was characterized by morphology, growth, and genomics with three giant indels and different types of HEs. The gene analysis on the required

  2. Genome assortment, not serogroup, defines Vibrio cholerae pandemic strains

    Energy Technology Data Exchange (ETDEWEB)

    Brettin, Thomas S [Los Alamos National Laboratory; Bruce, David C [Los Alamos National Laboratory; Challacombe, Jean F [Los Alamos National Laboratory; Detter, John C [Los Alamos National Laboratory; Han, Cliff S [Los Alamos National Laboratory; Munik, A C [Los Alamos National Laboratory; Chertkov, Olga [Los Alamos National Laboratory; Meincke, Linda [Los Alamos National Laboratory; Saunders, Elizabeth [Los Alamos National Laboratory; Choi, Seon Y [SEOUL NATL. UNIV.; Haley, Bradd J [U. MARYLAND; Taviani, Elisa [U. MARYLAND; Jeon, Yoon - Seong [INTL. VACCINE INST. SEOUL; Kim, Dong Wook [INTL. VACCINE INST. SEOUL; Lee, Jae - Hak [SEOUL NATL. UNIV.; Walters, Ronald A [PNNL; Hug, Anwar [NATL. INST. CHOLERIC ENTERIC DIS.; Colwell, Rita R [U. MARYLAND

    2009-01-01

    Vibrio cholerae, the causative agent of cholera, is a bacterium autochthonous to the aquatic environment, and a serious public health threat. V. cholerae serogroup O1 is responsible for the previous two cholera pandemics, in which classical and El Tor biotypes were dominant in the 6th and the current 7th pandemics, respectively. Cholera researchers continually face newly emerging and re-emerging pathogenic clones carrying combinations of new serogroups as well as of phenotypic and genotypic properties. These genotype and phenotype changes have hampered control of the disease. Here we compare the complete genome sequences of 23 strains of V. cholerae isolated from a variety of sources and geographical locations over the past 98 years in an effort to elucidate the evolutionary mechanisms governing genetic diversity and genesis of new pathogenic clones. The genome-based phylogeny revealed 12 distinct V. cholerae phyletic lineages, of which one, designated the V. cholerae core genome (CG), comprises both O1 classical and EI Tor biotypes. All 7th pandemic clones share nearly identical gene content, i.e., the same genome backbone. The transition from 6th to 7th pandemic strains is defined here as a 'shift' between pathogenic clones belonging to the same O1 serogroup, but from significantly different phyletic lineages within the CG clade. In contrast, transition among clones during the present 7th pandemic period can be characterized as a 'drift' between clones, differentiated mainly by varying composition of laterally transferred genomic islands, resulting in emergence of variants, exemplified by V.cholerae serogroup O139 and V.cholerae O1 El Tor hybrid clones that produce cholera toxin of classical biotype. Based on the comprehensive comparative genomics presented in this study it is concluded that V. cholerae undergoes extensive genetic recombination via lateral gene transfer, and, therefore, genome assortment, not serogroup, should be used to

  3. RpoS controls the Vibrio cholerae mucosal escape response.

    Directory of Open Access Journals (Sweden)

    Alex Toftgaard Nielsen

    2006-10-01

    Full Text Available Vibrio cholerae causes a severe diarrhoeal disease by secreting a toxin during colonization of the epithelium in the small intestine. Whereas the initial steps of the infectious process have been intensively studied, the last phases have received little attention. Confocal microscopy of V. cholerae O1-infected rabbit ileal loops captured a distinctive stage in the infectious process: 12 h post-inoculation, bacteria detach from the epithelial surface and move into the fluid-filled lumen. Designated the "mucosal escape response," this phenomenon requires RpoS, the stationary phase alternative sigma factor. Quantitative in vivo localization assays corroborated the rpoS phenotype and showed that it also requires HapR. Expression profiling of bacteria isolated from ileal loop fluid and mucus demonstrated a significant RpoS-dependent upregulation of many chemotaxis and motility genes coincident with the emigration of bacteria from the epithelial surface. In stationary phase cultures, RpoS was also required for upregulation of chemotaxis and motility genes, for production of flagella, and for movement of bacteria across low nutrient swarm plates. The hapR mutant produced near-normal numbers of flagellated cells, but was significantly less motile than the wild-type parent. During in vitro growth under virulence-inducing conditions, the rpoS mutant produced 10- to 100-fold more cholera toxin than the wild-type parent. Although the rpoS mutant caused only a small over-expression of the genes encoding cholera toxin in the ileal loop, it resulted in a 30% increase in fluid accumulation compared to the wild-type. Together, these results show that the mucosal escape response is orchestrated by an RpoS-dependent genetic program that activates chemotaxis and motility functions. This may furthermore coincide with reduced virulence gene expression, thus preparing the organism for the next stage in its life cycle.

  4. Bovine Lactoferrin and Lactoferrin-Derived Peptides Inhibit the Growth of Vibrio cholerae and Other Vibrio species

    Science.gov (United States)

    Acosta-Smith, Erika; Viveros-Jiménez, Karina; Canizalez-Román, Adrian; Reyes-Lopez, Magda; Bolscher, Jan G. M.; Nazmi, Kamran; Flores-Villaseñor, Hector; Alapizco-Castro, Gerardo; de la Garza, Mireya; Martínez-Garcia, Jesús J.; Velazquez-Roman, Jorge; Leon-Sicairos, Nidia

    2018-01-01

    Vibrio is a genus of Gram-negative bacteria, some of which can cause serious infectious diseases. Vibrio infections are associated with the consumption of contaminated food and classified in Vibrio cholera infections and non-cholera Vibrio infections. In the present study, we investigate whether bovine lactoferrin (bLF) and several synthetic peptides corresponding to bLF sequences, are able to inhibit the growth or have bactericidal effect against V. cholerae and other Vibrio species. The antibacterial activity of LF and LF-peptides was assessed by kinetics of growth or determination of colony forming unit in bacteria treated with the peptides and antibiotics. To get insight in the mode of action, the interaction between bLF and bLF-peptides (coupled to FITC) and V. cholera was evaluated. The damage of effector-induced bacterial membrane permeability was measured by inclusion of the fluorescent dye propidium iodide using flow cytometry, whereas the bacterial ultrastructural damage in bacteria treated was observed by transmission electron microscopy. The results showed that bLF and LFchimera inhibited the growth of the V. cholerae strains; LFchimera permeabilized the bacteria which membranes were seriously damaged. Assays with a multidrug-resistant strain of Vibrio species indicated that combination of sub-lethal doses of LFchimera with ampicillin or tetracycline strongly reduced the concentration of the antibiotics to reach 95% growth inhibition. Furthermore, LFchimera were effective to inhibit the V. cholerae counts and damage due to this bacterium in a model mice. These data suggest that LFchimera and bLF are potential candidates to combat the V. cholerae and other multidrug resistant Vibrio species. PMID:29375503

  5. Bovine Lactoferrin and Lactoferrin-Derived Peptides Inhibit the Growth of Vibrio cholerae and Other Vibrio species.

    Science.gov (United States)

    Acosta-Smith, Erika; Viveros-Jiménez, Karina; Canizalez-Román, Adrian; Reyes-Lopez, Magda; Bolscher, Jan G M; Nazmi, Kamran; Flores-Villaseñor, Hector; Alapizco-Castro, Gerardo; de la Garza, Mireya; Martínez-Garcia, Jesús J; Velazquez-Roman, Jorge; Leon-Sicairos, Nidia

    2017-01-01

    Vibrio is a genus of Gram-negative bacteria, some of which can cause serious infectious diseases. Vibrio infections are associated with the consumption of contaminated food and classified in Vibrio cholera infections and non-cholera Vibrio infections. In the present study, we investigate whether bovine lactoferrin (bLF) and several synthetic peptides corresponding to bLF sequences, are able to inhibit the growth or have bactericidal effect against V. cholerae and other Vibrio species. The antibacterial activity of LF and LF-peptides was assessed by kinetics of growth or determination of colony forming unit in bacteria treated with the peptides and antibiotics. To get insight in the mode of action, the interaction between bLF and bLF-peptides (coupled to FITC) and V. cholera was evaluated. The damage of effector-induced bacterial membrane permeability was measured by inclusion of the fluorescent dye propidium iodide using flow cytometry, whereas the bacterial ultrastructural damage in bacteria treated was observed by transmission electron microscopy. The results showed that bLF and LFchimera inhibited the growth of the V. cholerae strains; LFchimera permeabilized the bacteria which membranes were seriously damaged. Assays with a multidrug-resistant strain of Vibrio species indicated that combination of sub-lethal doses of LFchimera with ampicillin or tetracycline strongly reduced the concentration of the antibiotics to reach 95% growth inhibition. Furthermore, LFchimera were effective to inhibit the V. cholerae counts and damage due to this bacterium in a model mice. These data suggest that LFchimera and bLF are potential candidates to combat the V. cholerae and other multidrug resistant Vibrio species.

  6. Anti-vibrio potentials of acetone and aqueous leaf extracts of ...

    African Journals Online (AJOL)

    Purpose: To evaluate the anti-vibrio potentials of acetone and aqueous leaf extracts of Ocimum gratissimum and determine its relevance in the treatment of vibrios infection. Methods: The agar-well diffusion method was used for screening the extracts for their anti-vibrio activity. Broth micro-dilution assay was used to ...

  7. Genetic heterogeneity among Vibrio alginolyticus strains, and design of a PCR-based identification method using gyrB gene sequence.

    Science.gov (United States)

    Bunpa, Supansa; Nishibuchi, Mitsuaki; Thawonsuwan, Jumroensri; Sermwittayawong, Natthawan

    2017-10-10

    Vibrio alginolyticus, a pathogen among humans and marine animals, is ubiquitous in marine environments. The aims of this study were to analyze the relationships between genetic diversity and origins, and to develop new primers based on the gyrB sequence to identify V. alginolyticus isolated from various sources. To determine the genetic diversity of this bacterium, an arbitrarily primed polymerase chain reaction (AP-PCR) technique was performed on 36 strains of V. alginolyticus isolated from diarrhea patients and from diseased marine animals and environments in southern Thailand. The results showed distinct DNA fingerprints of all strains, indicating that they are genetically heterogeneous. For species-specific identification of V. alginolyticus, primers targeting the gyrB gene of V. alginolyticus were developed. Thirty reference Vibrio spp., 13 non-Vibrio spp., and 160 strains of V. alginolyticus isolated from various sources in southern Thailand were used to evaluate the specificity of these primers. Our results showed that the gyrB primers could specifically identify V. alginolyticus from all sample types. In addition, the detection limit of the PCR was at least 95 pg of DNA template. Therefore, we concluded that the newly designed gyrB primers are rapid, highly sensitive, and specific to identify V. alginolyticus isolated from various sources.

  8. ORF Alignment: NC_004459 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ase ... [Vibrio vulnificus CMCP6] ... Length = 153 ... Query: 124 DIHEGIEDTLVIFENRLKLHKLTTHYCSLPPIYCQPIALQQVWTNMISNALDAMP...EHGTL 183 ... DIHEGIEDTLVIFENRLKLHKLTTHYCSLPPIYCQPIALQQVWTNMISNALDAMP...EHGTL Sbjct: 1 ... DIHEGIEDTLVIFENRLKLHKLTTHYCSLPPIYCQPIALQQVWTNMISNALDAMPEHGTL 60 ... Query:

  9. ORF Alignment: NC_005139 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available nase ... [Vibrio vulnificus YJ016] ... Length = 154 ... Query: 494 ADIHEGIEDTLVIFENRLKLHKLTTHYCSLPPIYCQPIALQQVWTNMISNALDAM...PEHGT 553 ... ADIHEGIEDTLVIFENRLKLHKLTTHYCSLPPIYCQPIALQQVWTNMISNALDAMP...EHGT Sbjct: 1 ... ADIHEGIEDTLVIFENRLKLHKLTTHYCSLPPIYCQPIALQQVWTNMISNALDAMPEHGT 60 ... Query:

  10. Structural characterization of the extracellular polysaccharide from Vibrio cholerae O1 El-Tor.

    Directory of Open Access Journals (Sweden)

    Fitnat Yildiz

    Full Text Available The ability to form biofilms is important for environmental survival, transmission, and infectivity of Vibrio cholerae, the causative agent of cholera in humans. To form biofilms, V. cholerae produces an extracellular matrix composed of proteins, nucleic acids and a glycoconjugate, termed Vibrio exopolysaccharide (VPS. Here, we present the data on isolation and characterization of the polysaccharide part of the VPS (VPS-PS, which has the following structure: -4-α-GulpNAcAGly3OAc-(1-4-β-D-Glcp-(1-4-α-Glcp-(1-4-α-D-Galp-(1- where α-D-Glc is partially (∼20% replaced with α-D-GlcNAc. α-GulNAcAGly is an amide between 2-acetamido-2-deoxy-α-guluronic acid and glycine. Apparently, the polysaccharide is bound to a yet unidentified component, which gives it high viscosity and completely suppresses any NMR signals belonging to the sugar chains of the VPS. The only reliable method to remove this component at present is a treatment of the whole glycoconjugate with concentrated hydrochloric acid.

  11. BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING

    Directory of Open Access Journals (Sweden)

    Milagro FERNÁNDEZ-DELGADO

    2016-01-01

    Full Text Available Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM. Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS, while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing.

  12. BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING.

    Science.gov (United States)

    Fernández-Delgado, Milagro; Rojas, Héctor; Duque, Zoilabet; Suárez, Paula; Contreras, Monica; García-Amado, M Alexandra; Alciaturi, Carlos

    2016-01-01

    Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM). Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS), while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing.

  13. Nigribactin, a Novel Siderophore from Vibrio nigripulchritudo, Modulates Staphylococcus aureus Virulence Gene Expression

    Directory of Open Access Journals (Sweden)

    Lone Gram

    2012-11-01

    Full Text Available Staphylococcus aureus is a serious human pathogen that employs a number of virulence factors as part of its pathogenesis. The purpose of the present study was to explore marine bacteria as a source of compounds that modulate virulence gene expression in S. aureus. During the global marine Galathea 3 expedition, a strain collection was established comprising bacteria that express antimicrobial activity against Vibrio anguillarum and/or Staphylococcus aureus. Within this collection we searched colony material, culture supernatants, and cell extracts for virulence modulating activity showing that 68 out of 83 marine bacteria (affiliated with the Vibrionaceae and Pseudoalteromonas sp. influenced expression of S. aureus hla encoding α-hemolysin toxin and/or spa encoding Protein A. The isolate that upon initial screening showed the highest degree of interference (crude ethyl acetate extract was a Vibrio nigripulchritudo. Extraction, purification and structural elucidation revealed a novel siderophore, designated nigribactin, which induces spa transcription. The effect of nigribactin on spa expression is likely to be independent from its siderophore activity, as another potent siderophore, enterobactin, failed to influence S. aureus virulence gene expression. This study shows that marine microorganisms produce compounds with potential use in therapeutic strategies targeting virulence rather than viability of human pathogens.

  14. Chemoreceptor VfcA mediates amino acid chemotaxis in Vibrio fischeri.

    Science.gov (United States)

    Brennan, Caitlin A; DeLoney-Marino, Cindy R; Mandel, Mark J

    2013-03-01

    Flagellar motility and chemotaxis by Vibrio fischeri are important behaviors mediating the colonization of its mutualistic host, the Hawaiian bobtail squid. However, none of the 43 putative methyl-accepting chemotaxis proteins (MCPs) encoded in the V. fischeri genome has been previously characterized. Using both an available transposon mutant collection and directed mutagenesis, we isolated mutants for 19 of these genes, and screened them for altered chemotaxis to six previously identified chemoattractants. Only one mutant was defective in responding to any of the tested compounds; the disrupted gene was thus named vfcA (Vibrio fischeri chemoreceptor A; locus tag VF_0777). In soft-agar plates, mutants disrupted in vfcA did not exhibit the serine-sensing chemotactic ring, and the pattern of migration in the mutant was not affected by the addition of exogenous serine. Using a capillary chemotaxis assay, we showed that, unlike wild-type V. fischeri, the vfcA mutant did not undergo chemotaxis toward serine and that expression of vfcA on a plasmid in the mutant was sufficient to restore the behavior. In addition to serine, we demonstrated that alanine, cysteine, and threonine are strong attractants for wild-type V. fischeri and that the attraction is also mediated by VfcA. This study thus provides the first insights into how V. fischeri integrates information from one of its 43 MCPs to respond to environmental stimuli.

  15. Sentinel Surveillance Detects Low Circulation of Vibrio cholerae Serotype Inaba in Haiti, 2011-2012.

    Science.gov (United States)

    Llanes, Rafael; Lazo, Alcides; Somarriba, Lorenzo; Mas, Pedro

    2015-07-01

    Over 700,000 cases of cholera were reported in Haiti between October 2010 and February 2015. In November 2011, the Cuban Medical Team serving in Haiti established a laboratory-supported sentinel surveillance system for cholera in 10 public hospitals (one in each of Haiti's 10 departments), to estimate the proportion of hospitalized patients with cholera and detect emergence of new Vibrio cholerae serotypes. Each month, the first ten stool samples collected from patients admitted with acute watery diarrhea were studied in all hospitals involved. Surveillance system findings from November 1, 2011, to October 30, 2012 showed that acute watery diarrhea was caused by V. cholerae serogroup O1 in 45.9% (210/458) of patients: Serotype Ogawa was found in 98.6% of this isolates (207/210) and serotype Inaba in 1.4% (3/210), indicating low circulation level of the latter in Haiti. Continuing laboratory sentinel surveillance of V. cholerae is needed to monitor the spread of the disease and prevent and contain outbreaks, particularly of new serotypes. It is important to ensure that these findings are systematically integrated with data available to MSPP from other surveillance sources. KEYWORDS Vibrio cholerae, serotype Inaba, serotype Ogawa, epidemiological surveillance, medical cooperation, Haiti, Cuba.

  16. Comparative Genomic Analysis of Two Vibrio toranzoniae Strains with Different Virulence Capacity Reveals Clues on Its Pathogenicity for Fish.

    Science.gov (United States)

    Lasa, Aide; Gibas, Cynthia J; Romalde, Jesús L

    2017-01-01

    Vibrio toranzoniae is a Gram-negative bacterium of the Splendidus clade within the Vibrio genus. V. toranzoniae was first isolated from healthy clams in Galicia (Spain) but recently was also identified associated to disease outbreaks of red conger eel in Chile. Experimental challenges showed that the Chilean isolates were able to produce fish mortalities but not the strains isolated from clams. The aim of the present study was to determine the differences at the genomic level between the type strain of the species (CECT 7225(T)) and the strain R17, isolated from red conger eel in Chile, which could explain their different virulent capacity. The genome-based comparison showed high homology between both strains but differences were observed in certain gene clusters that include some virulence factors. Among these, we found that iron acquisition systems and capsule synthesis genes were the main differential features between both genomes that could explain the differences in the pathogenicity of the strains. Besides, the studied genomes presented genomic islands and toxins, and the R17 strain presented CRISPR sequences that are absent on the type strain. Taken together, this analysis provided important insights into virulence factors of V. toranzoniae that will lead to a better understanding of the pathogenic process.

  17. Vibrio cholerae pathogen from the freshwater-cultured whiteleg shrimp Penaeus vannamei and control with Bdellovibrio bacteriovorus.

    Science.gov (United States)

    Cao, Haipeng; An, Jian; Zheng, Weidong; He, Shan

    2015-09-01

    Vibriosis has become a major global economic problem in freshwater-farmed whiteleg shrimp (Penaeus vannamei). The prevention and control of vibriosis are now priority research topics. In this study, a pathogenic strain (QH) was isolated from vibriosis-infected freshwater-farmed P. vannamei that resulted in leg yellowing and was identified as a Vibrio cholerae isolate through phylogenetic analysis and the API 32GN system. A phylogenetic tree that was constructed using the neighbor-joining method further confirmed the QH isolate as a V. cholerae strain. A virulent outer membrane protein (ompU) gene was found to be present in the QH isolate, which further confirmed its pathogenicity. In addition, Bdellovibrio bacteriovorus conferred significant protection against V. cholerae: B. bacteriovorus exhibited significant bacteriolytic effects on the V. cholerae pathogen, possessed a wide prey range that included Vibrio pathogens, and displayed a positive protective efficacy against experimental V. cholerae infection in P. vannamei. To the best of our knowledge, this is the first report of the control of shrimp pathogen V. cholerae with B. bacteriovorus. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Development of a loop-mediated Isothermal amplification assay for sensitive and rapid detection of Vibrio parahaemolyticus

    Directory of Open Access Journals (Sweden)

    Kawahara Ryuji

    2008-09-01

    Full Text Available Abstract Background Vibrio parahaemolyticus is a marine seafood-borne pathogen causing gastrointestinal disorders in humans. Thermostable direct hemolysin (TDH and TDH-related hemolysin (TRH are known as major virulence determinants of V. parahaemolyticus. Most V. parahaemolyticus isolates from the environment do not produce TDH or TRH. Total V. parahaemolyticus has been used as an indicator for control of seafood contamination toward prevention of infection. Detection of total V. parahaemolyticus using conventional culture- and biochemical-based assays is time-consuming and laborious, requiring more than three days. Thus, we developed a novel and highly specific loop-mediated isothermal amplification (LAMP assay for the sensitive and rapid detection of Vibrio parahaemolyticus. Results The assay provided markedly more sensitive and rapid detection of V. parahaemolyticus strains than conventional biochemical and PCR assays. The assay correctly identified 143 V. parahaemolyticus strains, but did not detect 33 non-parahaemolyticus Vibrio and 56 non-Vibrio strains. Sensitivity of the LAMP assay for direct detection of V. parahaemolyticus in pure cultures and in spiked shrimp samples was 5.3 × 102 CFU per ml/g (2.0 CFU per reaction. The sensitivity of the LAMP assay was 10-fold more sensitive than that of the conventional PCR assay. The LAMP assay was markedly faster, requiring for amplification 13–22 min in a single colony on TCBS agar from each of 143 V. parahaemolyticus strains and less than 35 min in spiked shrimp samples. The LAMP assay for detection of V. parahaemolyticus required less than 40 min in a single colony on thiosulfate citrate bile salt sucrose (TCBS agar and 60 min in spiked shrimp samples from the beginning of DNA extraction to final determination. Conclusion The LAMP assay is a sensitive, rapid and simple tool for the detection of V. parahaemolyticus and will facilitate the surveillance for control of contamination of V

  19. Surface-attachment sequence in Vibrio Cholerae

    Science.gov (United States)

    Utada, Andrew; Gibiansky, Maxsim; Wong, Gerard

    2013-03-01

    Vibrio cholerae is a gram-negative bacterium that causes the human disease cholera. It is found natively in brackish costal waters in temperate climates, where it attaches to the surfaces of a variety of different aquatic life. V. cholerae has a single polar flagellum making it highly motile, as well as a number of different pili types, enabling it to attach to both biotic and abiotic surfaces. Using in-house built tracking software we track all surface-attaching bacteria from high-speed movies to examine the early-time attachment profile of v. cholerae onto a smooth glass surface. Similar to previous work, we observe right-handed circular swimming trajectories near surfaces; however, in addition we see a host of distinct motility mechanisms that enable rapid exploration of the surface before forming a more permanent attachment. Using isogenic mutants we show that the motility mechanisms observed are due to a complex combination of hydrodynamics and pili-surface interactions. Lauga, E., DiLuzio, W. R., Whitesides, G. M., Stone, H. A. Biophys. J. 90, 400 (2006).

  20. [Influence of aflatoxin on Vibrio fischeri luminescence].

    Science.gov (United States)

    Li, Xiang; Pan, Li; Wang, Bin

    2011-12-01

    In the present study, we aim to evaluate the inhibitory effect of aflatoxin on Vibrio fischeri luminescence. V. fischeri culture is treated with aflatoxin or the culture broth of aflatoxin-producing strains, and the luminescence intensity of V. fischeri is detected to analyze the influence of aflatoxin on V. fischeri. The logarithmic value of aflatoxin concentration and the decrease ratio of V. fischeri luminescence is in a linear relationship. Based on the regression equation between aflatoxin concentration and luminescence decrease of V. fischeri, the toxin-producing status of different microbes can be detected quickly and exactly: all of six tested Aspergillus flavus strains show toxigenicity to V. fischeri, and their toxin yield reached 14.94 mg/L - 46.45 mg/L (represented by aflatoxin concentration), while the tested Aspergillus oryzae shows no toxigenicity. The above data showed that the luminescence change of V. fischeri could exactly reflect the capability of various microbes to produce toxin (especially aflatoxin), which provided a new clue for rapid detection of aflatoxin in industrial and agricultural production and could be developed as a potential method for aflatoxin assay.

  1. Vibrio parahaemolyticus- An emerging foodborne pathogen

    Directory of Open Access Journals (Sweden)

    S Nelapati

    2012-02-01

    Full Text Available Vibrio parahaemolyticus is a halophilic gram negative, motile, oxidase positive, straight or curved rod-shaped, facultative anaerobic bacteria that occur naturally in the marine environment. They form part of the indigenous microflora of aquatic habitats of various salinity and are the major causative agents for some of the most serious diseases in fish, shellfish and penacid shrimp. This human pathogen causes acute gastroenteritis characterized by diarrhea, vomiting and abdominal cramps through consumption of contaminated raw fish or shellfish. V. parahaemolyticus is the leading cause of gastroenteritis due to the consumption of seafood worldwide. The incidence of V. parahaemolyticus infection has been increasing in many parts of the world, due to the emergence of O3:K6 serotype carrying the tdh gene which is responsible for most outbreaks worldwide. The pathogenicity of this organism is closely correlated with the Kanagawa phenomenon (KP + due to production of Kanagawa hemolysin or the thermostable direct hemolysin (TDH. The TDH and TRH (TDH-related hemolysin encoded by tdh and trh genes are considered to be important virulence factors. [Vet. World 2012; 5(1.000: 48-63

  2. Household Transmission of Vibrio cholerae in Bangladesh.

    Science.gov (United States)

    Sugimoto, Jonathan D; Koepke, Amanda A; Kenah, Eben E; Halloran, M Elizabeth; Chowdhury, Fahima; Khan, Ashraful I; LaRocque, Regina C; Yang, Yang; Ryan, Edward T; Qadri, Firdausi; Calderwood, Stephen B; Harris, Jason B; Longini, Ira M

    2014-11-01

    Vibrio cholerae infections cluster in households. This study's objective was to quantify the relative contribution of direct, within-household exposure (for example, via contamination of household food, water, or surfaces) to endemic cholera transmission. Quantifying the relative contribution of direct exposure is important for planning effective prevention and control measures. Symptom histories and multiple blood and fecal specimens were prospectively collected from household members of hospital-ascertained cholera cases in Bangladesh from 2001-2006. We estimated the probabilities of cholera transmission through 1) direct exposure within the household and 2) contact with community-based sources of infection. The natural history of cholera infection and covariate effects on transmission were considered. Significant direct transmission (p-valuelevels of pre-existing immunity to cholera infection. Transmission via direct exposure occurring outside of the household was not considered. Direct exposure contributes substantially to endemic transmission of symptomatic cholera in an urban setting. We provide the first estimate of the transmissibility of endemic cholera within prospectively-followed members of households. The role of direct transmission must be considered when planning cholera control activities.

  3. Role of Vibrio polysaccharide (vps) genes in VPS production, biofilm formation and Vibrio cholerae pathogenesis.

    Science.gov (United States)

    Fong, Jiunn C N; Syed, Khalid A; Klose, Karl E; Yildiz, Fitnat H

    2010-09-01

    Biofilm formation enhances the survival and persistence of the facultative human pathogen Vibrio cholerae in natural ecosystems and its transmission during seasonal cholera outbreaks. A major component of the V. cholerae biofilm matrix is the Vibrio polysaccharide (VPS), which is essential for development of three-dimensional biofilm structures. The vps genes are clustered in two regions, the vps-I cluster (vpsU, vpsA-K, VC0916-27) and the vps-II cluster (vpsL-Q, VC0934-39), separated by an intergenic region containing the rbm gene cluster that encodes biofilm matrix proteins. In-frame deletions of the vps clusters and genes encoding matrix proteins drastically altered biofilm formation phenotypes. To determine which genes within the vps gene clusters are required for biofilm formation and VPS synthesis, we generated in-frame deletion mutants for all the vps genes. Many of these mutants exhibited reduced capacity to produce VPS and biofilms. Infant mouse colonization assays revealed that mutants lacking either vps clusters or rbmA (encoding secreted matrix protein RbmA) exhibited a defect in intestinal colonization compared to the wild-type. Understanding the roles of the various vps gene products will aid in the biochemical characterization of the VPS biosynthetic pathway and elucidate how vps gene products contribute to VPS biosynthesis, biofilm formation and virulence in V. cholerae.

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

    Science.gov (United States)

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

    2013-12-01

    Clam Ruditapes philippinarum is one of the important marine aquaculture species in North China. However, pathogens can often cause diseases and lead to massive mortalities and economic losses of clam. In this work, we compared the metabolic responses induced by Vibrio anguillarum and Vibrio splendidus challenges towards hepatopancreas of clam using NMR-based metabolomics. Metabolic responses suggested that both V. anguillarum and V. splendidus induced disturbances in energy metabolism and osmotic regulation, oxidative and immune stresses with different mechanisms, as indicated by correspondingly differential metabolic biomarkers (e.g., amino acids, ATP, glucose, glycogen, taurine, betaine, choline and hypotaurine) and altered mRNA expression levels of related genes including ATP synthase, ATPase, glutathione peroxidase, heat shock protein 90, defensin and lysozyme. However, V. anguillarum caused more severe oxidative and immune stresses in clam hepatopancreas than V. splendidus. Our results indicated that metabolomics could be used to elucidate the biological effects of pathogens to the marine clam R. philippinarum. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Vibrio fischeri flavohaemoglobin protects against nitric oxide during initiation of the squid-Vibrio symbiosis.

    Science.gov (United States)

    Wang, Yanling; Dunn, Anne K; Wilneff, Jacqueline; McFall-Ngai, Margaret J; Spiro, Stephen; Ruby, Edward G

    2010-11-01

    Nitric oxide (NO) is implicated in a wide range of biological processes, including innate immunity against pathogens, signal transduction and protection against oxidative stress. However, its possible roles in beneficial host-microbe associations are less well recognized. During the early stages of the squid-vibrio symbiosis, the bacterial symbiont Vibrio fischeri encounters host-derived NO, which has been hypothesized to serve as a specificity determinant. We demonstrate here that the flavohaemoglobin, Hmp, of V. fischeri protects against NO, both in culture and during colonization of the squid host. Transcriptional analyses indicate that hmp expression is highly responsive to NO, principally through the repressor, NsrR. Hmp protects V. fischeri from NO inhibition of aerobic respiration, and removes NO under both oxic and anoxic conditions. A Δhmp mutant of V. fischeri initiates squid colonization less effectively than wild type, but is rescued by the presence of an NO synthase inhibitor. The hmp promoter is activated during the initial stage of colonization, during which the Δhmp strain fails to form normal-sized aggregates of colonizing cells. Taken together, these results suggest that the sensing of host-derived NO by NsrR, and the subsequent removal of NO by Hmp, influence aggregate size and, thereby, V. fischeri colonization efficiency. © 2010 Blackwell Publishing Ltd.

  6. Role for cheR of Vibrio fischeri in the Vibrio-squid symbiosis.

    Science.gov (United States)

    Deloney-Marino, Cindy R; Visick, Karen L

    2012-01-01

    Upon hatching, the Hawaiian squid Euprymna scolopes is rapidly colonized by its symbiotic partner, the bioluminescent marine bacterium Vibrio fischeri . Vibrio fischeri cells present in the seawater enter the light organ of juvenile squid in a process that requires bacterial motility. In this study, we investigated the role chemotaxis may play in establishing this symbiotic colonization. Previously, we reported that V. fischeri migrates toward numerous attractants, including N-acetylneuraminic acid (NANA), a component of squid mucus. However, whether or not migration toward an attractant such as squid-derived NANA helps the bacterium to localize toward the light organ is unknown. When tested for the ability to colonize juvenile squid, a V. fischeri chemotaxis mutant defective for the methyltransferase CheR was outcompeted by the wild-type strain in co-inoculation experiments, even when the mutant was present in fourfold excess. Our results suggest that the ability to perform chemotaxis is an advantage during colonization, but not essential.

  7. Reclassification of the larval pathogen for marine bivalves Vibrio tubiashii subsp. europaeus as Vibrio europaeus sp. nov.

    Science.gov (United States)

    Dubert, Javier; Romalde, Jesús L; Spinard, Edward J; Nelson, David R; Gomez-Chiarri, Marta; Barja, Juan L

    2016-11-01

    The Orientalis clade has a relevant significance for bivalve aquaculture since it includes the pathogens Vibrio bivalvicida, Vibrio tubiashii subsp. tubiashii and Vibrio tubiashii subsp. europaeus. However, the previous taxonomic description of the subspecies of V. tubiashii shows some incongruities that should be emended. In the genomic age, the comparison between genome assemblies is the key to clarify the taxonomic position of both subspecies. With this purpose, we have tested the ability of multilocus sequence analysis based on eight housekeeping gene sequences (gapA, gyrB, ftsZ, mreB, pyrH, recA, rpoA and topA), different in silico genome-to-genome comparisons, chemotaxonomic features and phenotypic traits to reclassify the subspecies V. tubiashii subsp. europaeus within the Orientalis clade. This polyphasic approach clearly demonstrated that this subspecies is phylogenetically and phenotypically distinct from V. tubiashii and should be elevated to the rank of species as Vibrio europaeus sp. nov. This reclassification allows us to update the Orientalis clade (V. bivalvicida,V. brasiliensis, V. crosai, V. hepatarius, V. orientalis, V. sinaloensis, V. tubiashii and V. europaeus sp. nov.) and reconstruct a better phylogeny of the genus Vibrio. An emended description of V. tubiashii is provided. Finally, the proposed novel species is represented by emergent bivalve pathogens [type strain PP-638T (=CECT 8136T=DSM 27349T), PP2-843 and 07/118 T2] responsible for high mortalities in Spanish and French hatcheries.

  8. Insights into bacteriophage application in controlling Vibrio species

    Directory of Open Access Journals (Sweden)

    Vengadesh Letchumanan

    2016-07-01

    Full Text Available Bacterial infections from various organisms including Vibrio sp. pose a serious hazard to humans in many forms from clinical infection to affecting the yield of agriculture and aquaculture via infection of livestock. Vibrio sp. is one of the main foodborne pathogens causing human infection and is also a common cause of losses in the aquaculture industry. Prophylactic and therapeutic usage of antibiotics has become the mainstay of managing this problem, however this in turn led to the emergence of multidrug resistant strains of bacteria in the environment; which has raised awareness of the critical need for alternative non antibiotic based methods of preventing and treating bacterial infections. Bacteriophages - viruses that infect and result in the death of bacteria – are currently of great interest as a highly viable alternative to antibiotics. This article provides an insight into bacteriophage application in controlling Vibrio species as well underlining the advantages and drawbacks of phage therapy.

  9. Reclassification of Vibrio fischeri, Vibrio logei, Vibrio salmonicida and Vibrio wodanis as Aliivibrio fischeri gen. nov., comb. nov., Aliivibrio logei comb. nov., Aliivibrio salmonicida comb. nov. and Aliivibrio wodanis comb. nov.

    Science.gov (United States)

    Urbanczyk, Henryk; Ast, Jennifer C; Higgins, Melissa J; Carson, Jeremy; Dunlap, Paul V

    2007-12-01

    Four closely related species, Vibrio fischeri, Vibrio logei, Vibrio salmonicida and Vibrio wodanis, form a clade within the family Vibrionaceae; the taxonomic status and phylogenetic position of this clade have remained ambiguous for many years. To resolve this ambiguity, we tested these species against other species of the Vibrionaceae for phylogenetic and phenotypic differences. Sequence identities for the 16S rRNA gene were > or =97.4 % among members of the V. fischeri group, but were Vibrio, with which they overlap in G+C content, and Enterovibrio, Grimontia and Salinivibrio, with which they do not overlap in G+C content). Combined analysis of the recA, rpoA, pyrH, gyrB and 16S rRNA gene sequences revealed that the species of the V. fischeri group form a tightly clustered clade, distinct from these other genera. Furthermore, phenotypic traits differentiated the V. fischeri group from other genera of the Vibrionaceae, and a panel of 13 biochemical tests discriminated members of the V. fischeri group from type strains of Photobacterium and Vibrio. These results indicate that the four species of the V. fischeri group represent a lineage within the Vibrionaceae that is distinct from other genera. We therefore propose their reclassification in a new genus, Aliivibrio gen. nov. Aliivibrio is composed of four species: Aliivibrio fischeri comb. nov. (the type species) (type strain ATCC 7744(T) =CAIM 329(T) =CCUG 13450(T) =CIP 103206(T) =DSM 507(T) =LMG 4414(T) =NCIMB 1281(T)), Aliivibrio logei comb. nov. (type strain ATCC 29985(T) =CCUG 20283(T) =CIP 104991(T) =NCIMB 2252(T)), Aliivibrio salmonicida comb. nov. (type strain ATCC 43839(T) =CIP 103166(T) =LMG 14010(T) =NCIMB 2262(T)) and Aliivibrio wodanis comb. nov. (type strain ATCC BAA-104(T) =NCIMB 13582(T) =LMG 24053(T)).

  10. Long-term effects of ocean warming on vibrios

    Science.gov (United States)

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

    2012-12-01

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

  11. Relative Frequency, Characteristics, and Antimicrobial Susceptibility Patterns of Vibrio spp., Aeromonas spp., Chromobacterium violaceum, and Shewanella spp. in the Northern Territory of Australia, 2000–2013

    Science.gov (United States)

    McAuliffe, Gary N.; Hennessy, Jann; Baird, Robert W.

    2015-01-01

    Vibrio, Aeromonas, Chromobacterium violaceum, and Shewanella (VACS) are water-associated Gram-negative organisms that can cause a variety of infections. The frequency, patient characteristics, and antimicrobial susceptibilities for 468 isolates from 442 patients from the Northern Territory were reviewed. Aeromonas spp. (312 of 468; 67%) were most commonly isolated followed by Vibrio spp. (71 of 468; 15%), Shewanella spp. (61 of 468; 13%), and C. violaceum (24 of 468; 5%). A strong male predominance was found (male to female ratio of 2.3:1). Skin and soft tissue isolations (373 of 468; 80%) from lower limb infections (222 of 371; 60%) were the most common clinical manifestation. The episodes were usually polymicrobial (281 of 468; 60%). Coisolates included Staphylococcus aureus (137 of 468; 29%), β-hemolytic streptococci (74 of 468; 16%), enterobacteriaceae (111 of 468; 24%), non-fermentative Gram-negative bacilli (35 of 468; 7%), and other VACS organisms (37 of 468; 8%). Antimicrobial resistance of VACS organisms to ciprofloxacin (0–4%), cefepime (0–3%), and gentamicin (0–0.8%) and Vibrio spp., Aeromonas spp., and Shewanella to cotrimoxazole (0–3%) was rarely shown. For water-associated lower limb skin and soft tissue infections in the tropics, clinicians should consider empirical antimicrobial therapy with agents active against S. aureus and VACS organisms. PMID:25548380

  12. Arabinose induces pellicle formation by Vibrio fischeri.

    Science.gov (United States)

    Visick, Karen L; Quirke, Kevin P; McEwen, Sheila M

    2013-03-01

    Biofilms are multicellular communities of bacteria attached to a surface and embedded in a protective matrix. In many cases, the signals that induce biofilm formation are unknown. Here, we report that biofilm formation by the marine bacterium Vibrio fischeri can be induced by the addition of arabinose to LBS (Luria-Bertani-salt), a tryptone-based medium. Growth of cells in the presence of 0.2% arabinose, but not other sugars, induced the production of a pellicle at the air/liquid interfaces of static cultures. V. fischeri failed to grow on arabinose as the sole carbon source, suggesting that pellicle production did not occur as a result of increased growth, but experiments using the acid/base indicator phenol red suggested that V. fischeri may partially metabolize arabinose. Pellicle production was independent of the syp polysaccharide locus but was altered upon disruption of the bcs cellulose locus. Through a screen for mutants defective for pellicle production, we found that loss of motility disrupted the formation of the arabinose-induced pellicle. Among the ∼20 mutants that retained motility were strains with insertions in a putative msh pilus locus and a strain with a defect in yidK, which is involved in galactose catabolism. Mutants with the msh gene disrupted grew poorly in the presence of arabinose, while the yidK mutant appeared to be "blind" to the presence of arabinose. Finally, arabinose impaired symbiotic colonization by V. fischeri. This work thus identifies a novel signal and new pathways involved in control of biofilm formation by V. fischeri.

  13. The light organ symbiont Vibrio fischeri possesses a homolog of the Vibrio cholerae transmembrane transcriptional activator ToxR.

    OpenAIRE

    Reich, K A; Schoolnik, G K

    1994-01-01

    A cross-hybridizing DNA fragment to Vibrio cholerae toxR was cloned from the nonpathogenic light organ symbiont Vibrio fischeri, and three proteins homologous to V. cholerae ToxR, ToxS, and HtpG were deduced from its DNA sequence. V. fischeri ToxR was found to activate a V. cholerae ToxR-regulated promoter, and an antiserum raised against the amino-terminal domain of V. cholerae ToxR cross-reacts V. fischeri ToxR.

  14. Structuration génétique d'une collection de souches de Vibrio. Contribution à la mise au point d'outils de typage moléculaire pour l'aide au diagnostic

    OpenAIRE

    Jaffres, Emmanuel

    2005-01-01

    Les bactéries du genre Vibrio représentent le groupe le plus fréquemment associé à des maladies infectieuses chez les mollusques marins. Ainsi chez l'huître creuse Crassostrea gigas, la majorité des Vibrios isolés d'animaux sains ou malades sont phénotypiquement classés dans un groupe de souches apparentées à Vibrio splendidus. Les récentes études phylogénétiques pars séquençage du gène gyrB, ont permis de distinguer plusieurs espèces au sein de ce groupe (V. splendidus, V. chagasii, V. cycli...

  15. Ocorrência de Vibrio parahaemolyticus em mexilhões (Perna perna, Linnaeus, 1758 de banco natural do litoral do município de Palhoça, Santa Catarina, Brasil Occurrence of Vibrio parahaemolyticus in mussels (Perna perna, Linnaeus, 1758 from a natural coastal bed in the municipality of Palhoça, Santa Catarina, Brazil

    Directory of Open Access Journals (Sweden)

    Rita Maria B. Archer

    1994-09-01

    Full Text Available Foi estudada a ocorrência de Vibrio parahaemolyticus em quarenta amostras de mexilhões (Perna perna, Linnaeus, 1758 procedentes de banco natural da praia da Pinheira, município de Palhoça, Santa catarina, durante um período de três meses. Na metodologia de isolamento foi utilizada a técnica do Número Mais Provável (NMP com enriquecimento em água alcalina peptonada e subseqüente plaqueamento em ágar TCBS (thiosulfate citrate bile salts sucrose. Em 52,5% das amostras de mexilhões foi constatada a presença de Vibrio parahaemolyticus, com níveis de contaminação entre A study was carried out on the occurrence of Vibrio parahaemolyticus in forty samples of mussels (Perna perna, Linnaeus, 1758 from a natural bed at Pinheira Beach, Municipality of Palhoça, Santa Catarina (Brazil, during a three-month period. The most probable number technique was used for isolation methodology with prior enrichment of samples in alkaline peptone water and subsequent planting on thiosulfate citrate bile salts sucrose agar. Vibrio parahaemolyticus was found in 52.5% of samples of mussels with a most probable number range of < 3 to 93 organisms/g. Serotyping was performed on 61 isolates and our findings indicate that 36.1% of these isolates were serologically non-typable; 54.1% of isolates displayed only flagellate antigenic structures and 8.2% had both antigenic structures. None of the isolates were Kanagawa positive.

  16. Vaccination in three different ways against vibriosis of Seriola dumerili caused by Vibrio hollisae

    Science.gov (United States)

    Ji, Rongxing; Zou, Wenzheng; Hu, Shiliu; Yan, Qingpi

    2008-08-01

    Bacterin was prepared by formalin-inactivating the virulent strain of Vibrio hollisae isolated from diseased Seriola dumerili (amberjack) suffering from vibriosis. Healthy S. dumerili were vaccinated by respective procedures of intramuscular injection, immersion, and orally administration. Results of the three different vaccinations were compared. Blood was drawn from the vaccinated fish every 7 days, and the antibody titers and lysozyme activities of the sera were determined. The antibody titer of injected fish was 1:40 at 7 d, and reached its peak of 1:320 at 28 d, while the fish vaccinated by immersion and orally administration exhibited weak antibody responses, the antibody titres of vaccinated fish exhibited significantly higher lysozyme activities ( Pvaccination of S. dumerili by the injection would be the best strategy to prevent the vibriosis in S. dumerili farm.

  17. Nigribactin, a Novel Siderophore from Vibrio nigripulchritudo, Modulates Staphylococcus aureus Virulence Gene Expression

    DEFF Research Database (Denmark)

    Nielsen, Anita; Månsson, Maria; Wietz, Matthias

    2012-01-01

    Staphylococcus aureus is a serious human pathogen that employs a number of virulence factors as part of its pathogenesis. The purpose of the present study was to explore marine bacteria as a source of compounds that modulate virulence gene expression in S. aureus. During the global marine Galathea...... 3 expedition, a strain collection was established comprising bacteria that express antimicrobial activity against Vibrio anguillarum and/or Staphylococcus aureus. Within this collection we searched colony material, culture supernatants, and cell extracts for virulence modulating activity showing...... that 68 out of 83 marine bacteria (affiliated with the Vibrionaceae and Pseudoalteromonas sp.) influenced expression of S. aureus hla encoding α-hemolysin toxin and/or spa encoding Protein A. The isolate that upon initial screening showed the highest degree of interference (crude ethyl acetate extract...

  18. Antagonistic Activity of Probiotic Organism Against Vibrio cholerae and Cryptococcus neoformans

    Directory of Open Access Journals (Sweden)

    Vidya, R.

    2010-01-01

    Full Text Available The microbes are useful in many ways in the modern world. Probiotics one of them, which refers to, acid adherence bacteria in the intestinal cells, are able to survive at low pH and produce large amount of lactic acid. The present investigation deals with the antagonistic activity of Lactobacillus acidophilus organism against pathogens. The organism was isolated from the curd sample. Identification of bacteria was done by various biochemical testing. The present study revealed that L. acidophilus inhibits Vibrio cholerae more efficiently than Streptococcus pneumoniae and Shigella dysentriae. When L. acidophilus and V. cholerae were grown together, L. acidophilus dominated the growth and competitively inhibited the growth of V. cholerae. L. acidophilus was also found to inhibit Cryptococcus neoformans.

  19. Detection of viable toxigenic Vibrio cholerae and virulent Shigella ...

    African Journals Online (AJOL)

    A rapid and sensitive assay was developed for the detection of low numbers of viable Vibrio cholerae and Shigella spp. cells in environmental and drinking water samples. Water samples were filtered, and the filters were enriched in a non-selective medium. The enrichment cultures were prepared for polymerase chain ...

  20. Molecular analysis of the emergence of pandemic Vibrio parahaemolyticus

    DEFF Research Database (Denmark)

    Boyd, EF; Cohen, AL; Naughton, LM

    2008-01-01

    Background Vibrio parahaemolyticus is abundant in the aquatic environment particularly in warmer waters and is the leading cause of seafood borne gastroenteritis worldwide. Prior to 1995, numerous V. parahaemolyticus serogroups were associated with disease, however, in that year an O3:K6 serogroup...

  1. Detection of quorum sensing molecules from Vibrio harveyi and use ...

    African Journals Online (AJOL)

    This paper explores the extraction and detection processes of quorum sensing molecules such as N-aceyl homoserine lactone compounds (AHL) from marine Vibrio harveyi. The spent culture of V. harveyi was solvent partitioned for AHL, rotary evaporated and re-suspended in 50% acetonitrile then detected with reporter ...

  2. Detection of viable toxigenic Vibrio cholerae and virulent Shigella ...

    African Journals Online (AJOL)

    DRINIE

    2003-04-02

    Apr 2, 2003 ... A rapid and sensitive assay was developed for the detection of low numbers of viable Vibrio cholerae and Shigella spp. cells in environmental and drinking water samples. Water samples were filtered, and the filters were enriched in a non-selective medium. The enrichment cultures were prepared for ...

  3. Survival of Vibrio cholerae in industrially polluted water, with ...

    African Journals Online (AJOL)

    containing industrial effluents. The effect of iron as well as pH on the survival of Vibrio cholerae (non-O1, El Tor and classical strains) in water samples from 12 points, where selected industrial effluents were discharged into rivers, was studied.

  4. Detection and confirmation of toxigenic Vibrio cholerae O1 in ...

    African Journals Online (AJOL)

    Epidemic cholera caused by toxigenic Vibrio cholerae O1 is a major health problem in several developing countries. Traditional methods for identifying V. cholerae involve cultural, biochemical and immunological assays which are cumbersome and often take several days to complete. In the present study, a direct cell ...

  5. Ion-swimming speed variation of Vibrio cholerae cells

    Indian Academy of Sciences (India)

    In the present work we report the variation in swimming speed of Vibrio cholerae with respect to the change in concentration of sodium ions in the medium. We have also studied the variation in swimming speed with respect to temperature. We find that the swimming speed initially shows a linear increase with the increase of ...

  6. Salmonella and Vibrio cholerae in Nile perch ( Lates niloticus ...

    African Journals Online (AJOL)

    The Nile perch (Lates niloticus) industry in East Africa has suffered severe economic losses in the last few years due to failure to comply with the microbiological standards of European Union (E.U). Fresh and frozen products have been suspected to be contaminated with Salmonella and Vibrio cholerae. This has led to a ...

  7. Detection and confirmation of toxigenic Vibrio cholerae O1 in ...

    African Journals Online (AJOL)

    2013-08-20

    Aug 20, 2013 ... Epidemic cholera caused by toxigenic Vibrio cholerae O1 is a major health problem in several developing countries. Traditional methods for identifying V. cholerae involve cultural, biochemical and immunological assays which are cumber- some and often take several days to complete. In the present study, ...

  8. antimicrobial susceptibility pattern of vibrio cholerae 01 strains

    African Journals Online (AJOL)

    hi-tech

    East African Medical Journal Vol. 77 No. 7 July 2000. ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF VIBRIO CHOLERAE 01 STRAINS DURING TWO CHOLERA OUTBREAKS IN DAR ES SALAAM,. TANZANIA. W.K. Urassa, MD, MSc, MMed, Lecturer, Department of Microbiology and Immunology, Muhimbili University ...

  9. Prevalence of Vibrio cholerae in rivers of Mpumalanga province ...

    African Journals Online (AJOL)

    Cholera is a life-threatening diarrhoeal disease, which mainly affects inhabitants of developing countries due to poor socio-economic conditions and lack of access to potable water and sanitation. Toxigenic Vibrio cholerae are the aetiological agents of cholera. These bacteria are autochthonous to aquatic environments, ...

  10. Vibriophages and Their Interactions with the Fish Pathogen Vibrio anguillarum

    DEFF Research Database (Denmark)

    Tan, Demeng; Gram, Lone; Middelboe, Mathias

    2014-01-01

    Vibrio anguillarum is an important pathogen in aquaculture, responsible for the disease vibriosis in many fish and invertebrate species. Disease control by antibiotics is a concern due to potential development and spread of antibiotic resistance. The use of bacteriophages to control the pathogen...

  11. Evaluation of in vitro Vibrio static activity of Shewanella algae ...

    African Journals Online (AJOL)

    conventional methods followed by Biolog microlog software. Since production of antagonistic agents rely on cultural conditions, antagonistic ability of candidate probioic against the mentioned Vibrios was assessed using Response Surface Methodology, with central composite design in which four independents variables ...

  12. Hybrid Vibrio cholerae El Tor lacking SXT identified as the cause of a cholera outbreak in the Philippines.

    Science.gov (United States)

    Klinzing, David C; Choi, Seon Young; Hasan, Nur A; Matias, Ronald R; Tayag, Enrique; Geronimo, Josefina; Skowronski, Evan; Rashed, Shah M; Kawashima, Kent; Rosenzweig, C Nicole; Gibbons, Henry S; Torres, Brian C; Liles, Veni; Alfon, Alicia C; Juan, Maria Luisa; Natividad, Filipinas F; Cebula, Thomas A; Colwell, Rita R

    2015-04-21

    Cholera continues to be a global threat, with high rates of morbidity and mortality. In 2011, a cholera outbreak occurred in Palawan, Philippines, affecting more than 500 people, and 20 individuals died. Vibrio cholerae O1 was confirmed as the etiological agent. Source attribution is critical in cholera outbreaks for proper management of the disease, as well as to control spread. In this study, three V. cholerae O1 isolates from a Philippines cholera outbreak were sequenced and their genomes analyzed to determine phylogenetic relatedness to V. cholerae O1 isolates from recent outbreaks of cholera elsewhere. The Philippines V. cholerae O1 isolates were determined to be V. cholerae O1 hybrid El Tor belonging to the seventh-pandemic clade. They clustered tightly, forming a monophyletic clade closely related to V. cholerae O1 hybrid El Tor from Asia and Africa. The isolates possess a unique multilocus variable-number tandem repeat analysis (MLVA) genotype (12-7-9-18-25 and 12-7-10-14-21) and lack SXT. In addition, they possess a novel 15-kb genomic island (GI-119) containing a predicted type I restriction-modification system. The CTXΦ-RS1 array of the Philippines isolates was similar to that of V. cholerae O1 MG116926, a hybrid El Tor strain isolated in Bangladesh in 1991. Overall, the data indicate that the Philippines V. cholerae O1 isolates are unique, differing from recent V. cholerae O1 isolates from Asia, Africa, and Haiti. Furthermore, the results of this study support the hypothesis that the Philippines isolates of V. cholerae O1 are indigenous and exist locally in the aquatic ecosystem of the Philippines. Genetic characterization and phylogenomics analysis of outbreak strains have proven to be critical for probing clonal relatedness to strains isolated in different geographical regions and over time. Recently, extensive genetic analyses of V. cholerae O1 strains isolated in different countries have been done. However, genome sequences of V. cholerae O1

  13. Isolation of putative probionts from cod rearing environment

    DEFF Research Database (Denmark)

    Lauzon, H.L.; Gudmundsdottir, S.; Pedersen, M.H.

    2008-01-01

    , metabolite production and adhesion to fish cell lines. Our study demonstrated that 14% of screened bacteria (n = 188) had antagonistic properties towards fish pathogens. The majority of these isolates were Gram-positive (81%), belonging to Firmicutes (69.2%) and Actinobacteria (11.5%) phyla based on 16S r......RNA gene sequencing. Only 6 (3.2%) of 188 isolates could inhibit all three pathogens tested: Vibrio anguillarum, Aeromonas salmonicida subsp. achromogenes and Vibrio salmonicida. Differences observed in activity intensity and spectrum among inhibitory isolates emphasise the need to develop probiotic...... mixtures for efficient prophylactic methods. Comparison of growth behaviour of inhibitory isolates and pathogens at cod rearing temperatures, metabolite production and adhesion capacity were considered for final probiont selection. Four promising isolates that could be used as a mixed supplement to rearing...

  14. Antimicrobial resistance in aerobic bacteria isolated from oral ...

    African Journals Online (AJOL)

    The species of bacteria detected included Bacillus spp, Pseudomonas spp, Staphylococcus spp, Streptococcus spp, Aeromonas spp, Burkholderia spp, Citrobacter spp, Escherichia spp, Enterobacter spp, Pasteurella spp, Burkholderia spp, Shewanella spp and Vibrio spp. Susceptibility of all identified isolates to ...

  15. Household Transmission of Vibrio cholerae in Bangladesh.

    Directory of Open Access Journals (Sweden)

    Jonathan D Sugimoto

    2014-11-01

    Full Text Available Vibrio cholerae infections cluster in households. This study's objective was to quantify the relative contribution of direct, within-household exposure (for example, via contamination of household food, water, or surfaces to endemic cholera transmission. Quantifying the relative contribution of direct exposure is important for planning effective prevention and control measures.Symptom histories and multiple blood and fecal specimens were prospectively collected from household members of hospital-ascertained cholera cases in Bangladesh from 2001-2006. We estimated the probabilities of cholera transmission through 1 direct exposure within the household and 2 contact with community-based sources of infection. The natural history of cholera infection and covariate effects on transmission were considered. Significant direct transmission (p-value<0.0001 occurred among 1414 members of 364 households. Fecal shedding of O1 El Tor Ogawa was associated with a 4.9% (95% confidence interval: 0.9%-22.8% risk of infection among household contacts through direct exposure during an 11-day infectious period (mean length. The estimated 11-day risk of O1 El Tor Ogawa infection through exposure to community-based sources was 2.5% (0.8%-8.0%. The corresponding estimated risks for O1 El Tor Inaba and O139 infection were 3.7% (0.7%-16.6% and 8.2% (2.1%-27.1% through direct exposure, and 3.4% (1.7%-6.7% and 2.0% (0.5%-7.3% through community-based exposure. Children under 5 years-old were at elevated risk of infection. Limitations of the study may have led to an underestimation of the true risk of cholera infection. For instance, available covariate data may have incompletely characterized levels of pre-existing immunity to cholera infection. Transmission via direct exposure occurring outside of the household was not considered.Direct exposure contributes substantially to endemic transmission of symptomatic cholera in an urban setting. We provide the first estimate of

  16. Multiple antibiotic resistance of Vibrio cholerae serogroup O139 in China from 1993 to 2009.

    Directory of Open Access Journals (Sweden)

    Li Yu

    Full Text Available Regarded as an emerging diarrheal micropathogen, Vibrio cholerae serogroup O139 was first identified in 1992 and has become an important cause of cholera epidemics over the last two decades. O139 strains have been continually isolated since O139 cholera appeared in China in 1993, from sporadic cases and dispersed foodborne outbreaks, which are the common epidemic types of O139 cholera in China. Antibiotic resistance profiles of these epidemic strains are required for development of clinical treatments, epidemiological studies and disease control. In this study, a comprehensive investigation of the antibiotic resistance of V. cholerae O139 strains isolated in China from 1993 to 2009 was conducted. The initial O139 isolates were resistant to streptomycin, trimethoprim-sulfamethoxazole and polymyxin B only, while multidrug resistance increased suddenly and became common in strains isolated after 1998. Different resistance profiles were observed in the isolates from different years. In contrast, most V. cholerae O1 strains isolated in the same period were much less resistant to these antibiotics and no obvious multidrug resistance patterns were detected. Most of the non-toxigenic strains isolated from the environment and seafood were resistant to four antibiotics or fewer, although a few multidrug resistant strains were also identified. These toxigenic O139 strains exhibited a high prevalence of the class I integron and the SXT element, which were rare in the non-toxigenic strains. Molecular subtyping of O139 strains showed highly diverse pulsed-field gel electrophoresis patterns, which may correspond to the epidemic state of sporadic cases and small-scale outbreaks and complex resistance patterns. Severe multidrug resistance, even resistance transfers based on mobile antibiotic resistance elements, increases the probability of O139 cholera as a threat to public health. Therefore, continual epidemiological and antibiotic sensitivity surveillance

  17. Presence of CTX gene cluster in environmental non-O1/O139 Vibrio cholerae and its potential clinical significance

    Directory of Open Access Journals (Sweden)

    B Bakhshi

    2012-01-01

    Full Text Available Purpose: The aim of this study was to understand the epidemiological linkage of clinical and environmental isolates of Vibrio cholerae and to determine their genotypes and virulence genes content. Materials and Methods: A total of 60 V. cholerae strains obtained from clinical specimens (n = 40 and surface waters (n = 20 were subjected to genotyping using PFGE and determination of their virulence-associated gene clusters. Result: PCR analysis showed the presence of chromosomally located hly and RTX genetic elements in 100% and 90% of the environmental isolates, respectively. The phage-mediated genetic elements such as CTX, TLC and VPI were detected in 5% of the environmental isolates suggesting that the environmental isolates cannot acquire certain mobile gene clusters. A total of 4 and 18 pulsotypes were obtained among the clinical and environmental V. cholerae isolates, respectively. Non-pathogenic environmentally isolated V. cholerae constituted a distinct cluster with one single non-O1, non-O139 strain (EP6 carrying the virulence genes similar to the epidemic strains. This may suggest the possible potential of conversion of non-pathogenic to a pathogenic environmental strain. Conclusions: The emergence of a single environmental isolate in our study containing the pathogenicity genes amongst the diverse non-pathogenic environmental isolates needs to be further studied in the context of V. cholerae pathogenicity sero-coversion.

  18. Stumbling across the Same Phage: Comparative Genomics of Widespread Temperate Phages Infecting the Fish Pathogen Vibrio anguillarum

    Directory of Open Access Journals (Sweden)

    Panos G. Kalatzis

    2017-05-01

    Full Text Available Nineteen Vibrio anguillarum-specific temperate bacteriophages isolated across Europe and Chile from aquaculture and environmental sites were genome sequenced and analyzed for host range, morphology and life cycle characteristics. The phages were classified as Siphoviridae with genome sizes between 46,006 and 54,201 bp. All 19 phages showed high genetic similarity, and 13 phages were genetically identical. Apart from sporadically distributed single nucleotide polymorphisms (SNPs, genetic diversifications were located in three variable regions (VR1, VR2 and VR3 in six of the phage genomes. Identification of specific genes, such as N6-adenine methyltransferase and lambda like repressor, as well as the presence of a tRNAArg, suggested a both mutualistic and parasitic interaction between phages and hosts. During short term phage exposure experiments, 28% of a V. anguillarum host population was lysogenized by the temperate phages and a genomic analysis of a collection of 31 virulent V. anguillarum showed that the isolated phages were present as prophages in >50% of the strains covering large geographical distances. Further, phage sequences were widely distributed among CRISPR-Cas arrays of publicly available sequenced Vibrios. The observed distribution of these specific temperate Vibriophages across large geographical scales may be explained by efficient dispersal of phages and bacteria in the marine environment combined with a mutualistic interaction between temperate phages and their hosts which selects for co-existence rather than arms race dynamics.

  19. Prevalence and distribution of Vibrio parahaemolyticus in finfish from Cochin (south India

    Directory of Open Access Journals (Sweden)

    Ammanamveetil A.M. Hatha

    2012-09-01

    Full Text Available Finfish samples obtained from four retail outlets in Cochin between June 2009 and June 2010 were investigated for the occurrence of Vibrio parahaemolyticus. A total of 182 samples were collected and suspect isolates were identified using standard biochemical tests and were further confirmed by a species-specific tlh gene. V. parahaemolyticus was detected in 45.1% of samples, with demersal fish being more affected than pelagic species. The bacterium was isolated more frequently from the skin and gills of pelagic fish, while the intestine yielded greater numbers of V. parahaemolyticus in demersal fish. The highest incidence of antibiotic resistance was recorded against ampicillin and streptomycin, followed by carbenicillin, cefpodoxime, cephalothin, colistin and amoxycillin; the lowest was against nalidixic acid, tetracycline, chloramphenicol and ciprofloxacin. Multiple drug resistance was prevalent among isolates. Although only a fraction of strains are pathogenic for humans, the time-temperature abuse in markets provides ample scope for these strains to multiply to dangerous levels. The multidrug resistant nature of the strains adds to the gravity of the problem. High V. parahaemolyticus incidence rates in market finfish samples from areas in and around Cochin clearly indicates that control measures should be adopted to reduce post-harvest contamination in seafood and time-temperature abuse in markets to diminish the risk of V. parahaemolyticus infection associated with seafood destined for human consumption.

  20. [A comparative analysis of genomes of virulent and avirulent strains of Vibrio cholerae O139].

    Science.gov (United States)

    Eroshenko, G A; Osin, A V; Shchelkanova, E Iu; Smirnova, N I

    2004-01-01

    A comparative analysis of the genome of V. cholerae O139 strains isolated in Russia's territory from patients with cholera and from the environment showed essential differences in their structures. The genome of clinical strains possessed all tested genes associated with virulence (ctxAB, zot, ace, rstC, rtxA, hap, toxR and toxT) and the at-tRS site for the CTXp phage DNA integration. As for the O139 V. cholerae chromosome strains isolated from water, 70% of the studied genes (ctxAB, zot, ace, rstC, tcpA, and toxT) and the attRS sequence were not detected in them. A lack of the key virulence genes in O139-serogroup "water" vibrios, including genes of toxin-coregulated adhesion pili. (that are receptors for the CTXp phage), and of the attachment site of the above phage are indicative of that the O139 V. cholerae strains isolated from open water sources located in different Russia's regions are epidemically negligible.

  1. Vibrio parahaemolyticus a causative bacterium for tail rot disease in ornamental fish, Amphiprion sebae

    Directory of Open Access Journals (Sweden)

    Thangapandi Marudhupandi

    2017-11-01

    Full Text Available The present study was performed to identify the tail rot disease causing bacterium in marine ornamental fish, Amphiprion sebae. Bacteria were isolated from the infected immune organs and tail region of A. sebae. Five different bacterial isolates (S1-S5 with different shape, size and colour were chosen for the infection study. The isolated strains were individually challenged with A. sebae at a constant dose of 1 × 107 CFU/fish. The virulent strain was found to be S-3, which showed maximum reproducing ability in A. sebae by causing typical tail rot disease and mortality. Furthermore, S-3 strain was identified as Vibrio parahaemolyticus by 16S rRNA gene sequencing (KF738005, biochemical analysis and amplification of tox R gene. Subsequently, extracellular products (ECPs of V. parahaemolyticus were prepared by cellophane overlay method. The LD50 value of V. parahaemolyticus and its ECPS were found to be 1 × 105 CFU and 5 μg/fish. The histology results revealed that V. parahaemolyticus and its ECPS are the major cause of tail rot disease in A. sebae.

  2. Isolation and antibiotic sensitivity of Aeromonas from children with diarrhea

    Directory of Open Access Journals (Sweden)

    Meiyanti Meiyanti

    2016-02-01

    Full Text Available Aeromonas species are gram-negative, motile, facultative anaerobic, rod shaped, oxidase positive bacteria of the recently assigned family Aeromonadaceae. The significance of Aeromonas species as causative agent of human diarrhoea has recently been established. The aim of the present study was to investigate the distribution, and antibiotic sensitivity of Aeromonas in nonhospitalized children with diarrhea.One hundred and seventeen rectal swabs from children with diarhhea were cultured for isolation of Aeromonas organisms as the etiological agents. In addition to Aeromonas, other enteric pathogens were also isolated. Overall, the isolates of enteric pathogens amounted to 36.8%, consisting of Salmonella, Shigella, Aeromonas, and Vibrio. Aeromonas was only found in 5.1% of cultures, with a ratio of A. caviae and A. hydrophila of 2:1, while Salmonella made up the majority of causative organisms with an isolation frequency of 18.8%, followed by Shigella with 11.1%. In this study no isolates of Vibrio cholerae O1 were found as etiological agents of diarrhea; however, V. cholerae non-O1 and V. parahaemolyticus were found in small numbers (<1%. All isolates of Aeromonas were resistant to ampicillin and tetracycline, but sensitive to ciprofloxacin and ceftriaxone, as were the other enteric pathogens. Although the frequency of isolation of these enteric pathogens was higher than for Vibrio spp., their role in infective diarrhea was less clearcut in comparison with Salmonella and Shigella.

  3. Evaluation of Cholera Toxin Expression in Different Populations of Vibrio cholera

    OpenAIRE

    Sedigheh Ebrahimi Kasgari; Mahnaz Nourani; Yousef Yahyapour; Seyed Ehsanollah Mousavi; Enayatollah Kalantar; Hami Kaboosi; Seyed Mahmoud Amin Marashi

    2015-01-01

    Background: Cholera is one of the most diseases of human. Cholera toxin is the most important pathogenic factor in humans that causes diarrhea. The cholera toxin is produced by V. cholerae and CTXфPhage. Objectives: In this study, we have investigated the production cholera toxin with different density of Vibrio cholerae. Materials and Methods: With this propose we inoculated classical strain O1 of Vibrio cholerae ATCC 14035 and Vibrio cholerae O1biovar El Tor N16961 into th...

  4. VibrioBase: A Model for Next-Generation Genome and Annotation Database Development

    Directory of Open Access Journals (Sweden)

    Siew Woh Choo

    2014-01-01

    Full Text Available To facilitate the ongoing research of Vibrio spp., a dedicated platform for the Vibrio research community is needed to host the fast-growing amount of genomic data and facilitate the analysis of these data. We present VibrioBase, a useful resource platform, providing all basic features of a sequence database with the addition of unique analysis tools which could be valuable for the Vibrio research community. VibrioBase currently houses a total of 252 Vibrio genomes developed in a user-friendly manner and useful to enable the analysis of these genomic data, particularly in the field of comparative genomics. Besides general data browsing features, VibrioBase offers analysis tools such as BLAST interfaces and JBrowse genome browser. Other important features of this platform include our newly developed in-house tools, the pairwise genome comparison (PGC tool, and pathogenomics profiling tool (PathoProT. The PGC tool is useful in the identification and comparative analysis of two genomes, whereas PathoProT is designed for comparative pathogenomics analysis of Vibrio strains. Both of these tools will enable researchers with little experience in bioinformatics to get meaningful information from Vibrio genomes with ease. We have tested the validity and suitability of these tools and features for use in the next-generation database development.

  5. Fasciitis necroticans ten gevolge van een infectie met in een palingkwekerij

    NARCIS (Netherlands)

    Dijkstra, A.; Ingen, van J.; Lubbert, P.H.W.; Haenen, O.L.M.; Möller, A.V.M.

    2009-01-01

    Een 41-jarige man presenteerde zich met een pijnlijke infectie van zijn rechter hand. Hij werkte in een palingkwekerij, waar op dat moment een infectie met Vibrio vulnificus onder de palingen heerste. Vibrio vulnificus als veroorzaker van een wondinfectie, een sepsis of een gastro-enteritis is een

  6. FURTHER STUDIES ON THE ETIOLOGICAL ROLE OF VIBRIO FETUS.

    Science.gov (United States)

    Smith, T; Little, R B; Taylor, M S

    1920-11-30

    The data bearing on these three cases are quite sufficient to rule out Bacillus abortus as the agent. Not only the cultures and guinea pig tests of fetal tissues and contents of the digestive tract, but also the agglutination and guinea pig tests of the milk, were negative. The same is true of the agglutination tests of the blood serum. Only in one case was the placenta obtained in part. The stained films and the sections from various regions showed no abortion bacilli. Guinea pig tests of placental tissue were negative for Bacillus abortus. On the other hand) minute organisms resembling vibrios were detected in the cytoplasm of endothelial cells within capillaries in the edematous subchorionic tissue. Subsequently the agglutination titer of the blood serum of one of these cases rose to a level indicating infection with Bacillus abortus during the second pregnancy. The peculiar distribution of abortions due to Vibrio fetus among older cows and heifers in this herd, resulting at first in cases among older cows and latterly passing to young stock, may be explained by certain occurrences in the herd itself. It may be assumed that the infection was originally brought in by purchased cows. The young stock is kept segregated from these in a special barn, and when 6 months old it is pastured on outlying farms until returned in an advanced stage of pregnancy. The heifers during the first pregnancy were thus kept away from vibrio carriers until after the first calf was born. In June and July, 1919, 55 older cows, purchased and native, were placed on the young stock pasture. The three cases of abortion in heifers due to Vibrio fetus occurred October 24, November 9, and December 2, 1919. The age and condition of the fetuses accord very well with the assumption that Vibrio fetus was introduced among the young stock in June or July of the same year. The information gathered thus far concerning vibrionic abortion in this herd enables us to formulate a tentative hypothesis

  7. Advanced Microbial Taxonomy Combined with Genome-Based-Approaches Reveals that Vibrio astriarenae sp. nov., an Agarolytic Marine Bacterium, Forms a New Clade in Vibrionaceae.

    Directory of Open Access Journals (Sweden)

    Nurhidayu Al-Saari

    Full Text Available Advances in genomic microbial taxonomy have opened the way to create a more universal and transparent concept of species but is still in a transitional stage towards becoming a defining robust criteria for describing new microbial species with minimum features obtained using both genome and classical polyphasic taxonomies. Here we performed advanced microbial taxonomies combined with both genome-based and classical approaches for new agarolytic vibrio isolates to describe not only a novel Vibrio species but also a member of a new Vibrio clade. Two novel vibrio strains (Vibrio astriarenae sp. nov. C7T and C20 showing agarolytic, halophilic and fermentative metabolic activity were isolated from a seawater sample collected in a coral reef in Okinawa. Intraspecific similarities of the isolates were identical in both sequences on the 16S rRNA and pyrH genes, but the closest relatives on the molecular phylogenetic trees on the basis of 16S rRNA and pyrH gene sequences were V. hangzhouensis JCM 15146T (97.8% similarity and V. agarivorans CECT 5085T (97.3% similarity, respectively. Further multilocus sequence analysis (MLSA on the basis of 8 protein coding genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA, and topA obtained by the genome sequences clearly showed the V. astriarenae strain C7T and C20 formed a distinct new clade protruded next to V. agarivorans CECT 5085T. The singleton V. agarivorans has never been included in previous MLSA of Vibrionaceae due to the lack of some gene sequences. Now the gene sequences are completed and analysis of 100 taxa in total provided a clear picture describing the association of V. agarivorans into pre-existing concatenated network tree and concluded its relationship to our vibrio strains. Experimental DNA-DNA hybridization (DDH data showed that the strains C7T and C20 were conspecific but were separated from all of the other Vibrio species related on the basis of both 16S rRNA and pyrH gene phylogenies (e.g., V

  8. Characterization of the secretomes of two vibrios pathogenic to mollusks.

    Science.gov (United States)

    Madec, Stéphanie; Pichereau, Vianney; Jacq, Annick; Paillard, Mathieu; Boisset, Claire; Guérard, Fabienne; Paillard, Christine; Nicolas, Jean-Louis

    2014-01-01

    Vibrio tapetis causes the brown ring disease in the Japanese clam Ruditapes philippinarum while Vibrio aestuarianus is associated with massive oyster mortalities. As extracellular proteins are often associated with the virulence of pathogenic bacteria, we undertook a proteomic approach to characterize the secretomes of both vibrios. The extracellular proteins (ECPs) of both species were fractionated by SEC-FPLC and in vitro assays were performed to measure the effects of each fraction on hemocyte cellular parameters (phagocytosis and adhesion). Fractions showing a significant effect were subjected to SDS-PAGE, and proteins were identified by nano LC-MS/MS. 45 proteins were identified for V. aestuarianus and 87 for V. tapetis. Most of them belonged to outer membrane or were periplasmic, including porins or adhesins that were already described as virulence factors in other bacterial species. Others were transporter components, flagella proteins, or proteins of unknown function (14 and 15 respectively). Interestingly, for V. aestuarianus, we noted the secretion of 3 extracellular enzymes including the Vam metalloprotease and two other enzymes (one putative lipase and one protease). For V. tapetis, we identified five extracellular enymes, i.e. two different endochitinases, one protease, one lipase and an adhesin. A comparison of both secretomes also showed that only the putative extracellular lipase was common to both secretomes, underscoring the difference in pathogenicity mechanisms between these two species. Overall, these results characterize for the first time the secretomes of these two marine pathogenic vibrios and constitute a useful working basis to further analyze the contribution of specific proteins in the virulence mechanisms of these species.

  9. Mutation of Bacterium Vibrio gazogenes for Selective Preparation of Colorants

    OpenAIRE

    Alihosseini, Farzaneh; Lango, Jozsef; Ju, Kou-San; Hammock, Bruce D.; Sun, Gang

    2010-01-01

    A novel marine bacterium strain effectively produced prodiginine type pigments. These colorants could dye wool, silk and synthetic fabrics such as polyester and polyacrylic and also show antibacterial properties against Escherichia coli and Staphylococcus aureus bacteria on the dyed products. Methyl nitrosoguanidine was used as a mutation agent to increase the genetic diversity and the production yield of the bacteria of the family of Vibrio gazogenes. The analysis of the mutated samples show...

  10. Wind direction and its linkage with Vibrio cholerae dissemination.

    Science.gov (United States)

    Paz, Shlomit; Broza, Meir

    2007-02-01

    The relevance of climatic events as causative factors for cholera epidemics is well known. However, examinations of the involvement of climatic factors in intracontinental disease distribution are still absent. The spreading of cholera epidemics may be related to the dominant wind direction over land. We examined the geographic diffusion of three cholera outbreaks through their linkage with the wind direction: a) the progress of Vibrio cholerae O1 biotype El Tor in Africa during 1970-1971 and b) again in 2005-2006; and c) the rapid spread of Vibrio cholerae O139 over India during 1992-1993. We also discuss the possible influence of the wind direction on windborn dissemination by flying insects, which may serve as vectors. Analysis of air pressure data at sea level and at several altitudes over Africa, India, and Bangladesh show a correspondence between the dominant wind direction and the intracontinental spread of cholera. We explored the hypothesis that winds have assisted the progress of cholera Vibrios throughout continents. The current analysis supports the hypothesis that aeroplankton (the tiny life forms that float in the air and that may be caught and carried upward by the wind, landing far from their origin) carry the cholera bacteria from one body of water to an adjacent one. This finding may improve our understanding of how climatic factors are involved in the rapid distribution of new strains throughout a vast continental area. Awareness of the aerial transfer of Vibrio cholerae may assist health authorities by improving the prediction of the disease's geographic dissemination.

  11. Adsorption kinetics of laterally and polarly flagellated Vibrio.

    OpenAIRE

    Belas, M R; Colwell, R R

    1982-01-01

    The adsorption of laterally and polarly flagellated bacteria to chitin was measured, and from the data obtained, a modified Langmuir adsorption isotherm was derived. Results indicated that the adsorption of laterally flagellated Vibrio parahaemolyticus follows the Langmuir adsorption isotherm, a type of adsorption referred to as surface saturation kinetics, when conditions are favorable for the production of lateral flagella. When conditions were not favorable for the production of lateral fl...

  12. Revealing the ability of a novel polysaccharide bioflocculant in bioremediation of heavy metals sensed in a Vibrio bioluminescence reporter assay.

    Science.gov (United States)

    Sajayan, Arya; Seghal Kiran, G; Priyadharshini, S; Poulose, Navya; Selvin, Joseph

    2017-09-01

    A bioflocculant-producing bacterial strain, designated MSI021, was isolated from the marine sponge Dendrilla nigra and demonstrated 94% flocculation activity in a kaolin clay suspension. MSI021 was identified as Bacillus cereus based on phylogenetic affiliation and biochemical characteristics. The purified extra-cellular bioflocculant was chemically elucidated as a polysaccharide molecule. The polysaccharide bioflocculant was stable under both acidic and alkaline conditions (pH 2.0-10.0) and temperatures up to 100 °C. The purified bioflocculant efficiently nucleated the formation of silver nanoparticles which showed broad spectrum antibacterial activity. The ability of the bioflocculant to remediate heavy metal toxicity was evaluated by measuring the inhibition of bioluminescence expression in Vibrio harveyi. Enrichment of heavy metals such as zinc, mercury and copper at concentrations of 1, 2 and 3 mM in culture media showed significant reduction of bioluminescence in Vibrio, whereas media enriched with heavy metals and bioflocculant showed dose dependent improvement in the expression of bioluminescence. The assay results demonstrated that the polysaccharide bioflocculant effectively mitigates heavy metal toxicity, thereby improving the expression of bioluminescence in Vibrio. This bioluminescence reporter assay can be developed into a high-throughput format to monitor and evaluate of heavy metal toxicity. The findings of this study revealed that a novel polysaccharide bioflocculant produced by a marine B. cereus demonstrated strong flocculating performance and was effective in nucleating the formation antibacterial silver nanoparticles and removing heavy metals. These results suggest that the MSI021 polysaccharide bioflocculant can be used to develop greener waste water treatment systems. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Biocompatible capped iron oxide nanoparticles for Vibrio cholerae detection

    Science.gov (United States)

    Sharma, Anshu; Baral, Dinesh; Rawat, Kamla; Solanki, Pratima R.; Bohidar, H. B.

    2015-05-01

    We report the studies relating to fabrication of an efficient immunosensor for Vibrio cholerae detection. Magnetite (iron oxide (Fe3O4)) nanoparticles (NPs) have been synthesized by the co-precipitation method and capped by citric acid (CA). These NPs were electrophoretically deposited onto indium-tin-oxide (ITO)-coated glass substrate and used for immobilization of monoclonal antibodies against Vibrio cholerae (Ab) and bovine serum albumin (BSA) for Vibrio cholerae detection using an electrochemical technique. The structural and morphological studies of Fe3O4 and CA-Fe3O4/ITO were characterized by x-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectroscopy, and dynamic light scattering (DLS) techniques. The average crystalline size of Fe3O4, CA-Fe3O4 nanoparticles obtained were about 29 ± 1 nm and 37 ± 1 nm, respectively. The hydrodynamic radius of the nanoparticles was found to be 77.35 nm (Fe3O4) and 189.51 nm (CA-Fe3O4) by DLS measurement. The results of electrochemical response studies of the fabricated BSA/Ab/CA-Fe2O3/ITO immunosensor exhibits a good detection range of 12.5-500 ng mL-1 with a low detection limit of 0.32 ng mL-1, sensitivity 0.03 Ω/ng ml-1 cm-2, and reproducibility more than 11 times.

  14. In vitro biofilm forming capacity on abiotic contact surfaces by outbreak-associated Vibrio harveyi strains

    Directory of Open Access Journals (Sweden)

    Pallaval Veera Bramha Chari

    2014-02-01

    Full Text Available Objective: To evaluate the in vitro biofilm forming capacity on abiotic food contact surfaces by Vibrio harveyi (V. harveyi strains. Methods: Thirty six Gram-negative V. harveyi strains were isolated from various street vended seafood outlets in a food processing line and evaluated for their ability to produce mucoid biofilms on food contact surfaces using a microplate assay. Phenotypic characterization of mucoid biofilm producing V. harveyi strains were screened on Congo red agar, thiosulfate-citrate-bile salts-sucrose agar and tryptic soy agar, respectively. Results: Only five V. harveyi strains (14% were mucoid biofilm producers characterized by formation of black colonies, whereas the remaining 31 strains (86% were not capable of producing biofilm characterized by formation of red colonies or pinkish-red colonies with darkening at the centre. The morphological, physiological and biochemical characteristics of these isolates were studied using standard protocols. Strain identification was confirmed by polymerase chain reaction targeted to species-specific polymerase chain reaction primers VH-1 and VH-2 corresponding to variable regions of V. harveyi 16S rRNA sequence. All the biofilm-forming strains showed resistance to at least three antimicrobial compounds tested. V. harveyi strains isolated from various seafood were able to form biofilms of different capacity, and the strains VB267, VB238 and VB166 isolated from cat fish, shrimp and eel fish exhibited significantly greater biofilm forming ability compared to other isolates. Conclusions: It can be concluded from the present study that the strain VB166 was able to better attach and form subsequent biofilms on glass and stainless steel compared to high density polyethylene. These properties allow these bacteria to survive, proliferate and persist in street vended seafood outlets.

  15. Pathogenicity profile of Vibrio parahaemolyticus in farmed Pacific white shrimp, Penaeus vannamei.

    Science.gov (United States)

    Ananda Raja, R; Sridhar, R; Balachandran, C; Palanisammi, A; Ramesh, S; Nagarajan, K

    2017-08-01

    A pathobiological study was conducted using Vibrio parahaemolyticus (VP) strain isolated from vibriosis affected shrimp (Penaeus vannamei) farms in Kancheepuram and Thiruvallur districts of Tamil Nadu during August 2014 to February 2015. The isolate was identified based on the morphological, physiological, biochemical and molecular characters. LD50 value with intramuscular injection was determined as 2.6 × 104 cfu/shrimp and sequential pathology was studied giving 6.1 × 103 cfu/shrimp (LD25). Total plate count (TPC) and total Vibrio count (TVC) in water, pond sediment, haemolymph, muscle, HP and gut were found significantly (P vannamei. Major histopathological manifestations observed were haemocyte infiltration/nodule in the epidermis, skeletal and cardiac muscles, atrophy of the excretory organ, and disrupted HP tubules with diffuse interstitial edema and haemocytic infiltration. Further HP showed that there was thickening of intertubular space, karyomegaly with prominent nucleoli, rounding and sloughing of HP tubular epithelium, many mitotic figures with bacterial colonies and apoptotic bodies, separation of shrunken tubule epithelium from myoepithelial fibers, regeneration of tubules, cystic, dilated and vacuolated appearance of HP tubules, hypoplastic changes in the tubules with no B, R and F cells, granuloma formation, concretions in tubules, calcification, necrosis, and washed out appearance with complete loss of architecture. The progression of the degenerative changes in the HP tubular epithelial cells was from proximal to distal end. In haematopoietic organ, increased mitotic activities with focal to extensive depletion and degeneration were observed. Degeneration of the stromal matrix with spheroid formation in lymphoid organ was observed among the Vp infected natural and experimental animals. Degeneration of glandular structures in the prehensile appendages with bacterial colonies, melanization and loss of epithelial layer in oesophagus

  16. Genome-Wide Biases in the Rate and Molecular Spectrum of Spontaneous Mutations in Vibrio cholerae and Vibrio fischeri.

    Science.gov (United States)

    Dillon, Marcus M; Sung, Way; Sebra, Robert; Lynch, Michael; Cooper, Vaughn S

    2017-01-01

    The vast diversity in nucleotide composition and architecture among bacterial genomes may be partly explained by inherent biases in the rates and spectra of spontaneous mutations. Bacterial genomes with multiple chromosomes are relatively unusual but some are relevant to human health, none more so than the causative agent of cholera, Vibrio cholerae Here, we present the genome-wide mutation spectra in wild-type and mismatch repair (MMR) defective backgrounds of two Vibrio species, the low-%GC squid symbiont V. fischeri and the pathogen V. cholerae, collected under conditions that greatly minimize the efficiency of natural selection. In apparent contrast to their high diversity in nature, both wild-type V. fischeri and V. cholerae have among the lowest rates for base-substitution mutations (bpsms) and insertion-deletion mutations (indels) that have been measured, below 10(-)(3)/genome/generation. Vibrio fischeri and V. cholerae have distinct mutation spectra, but both are AT-biased and produce a surprising number of multi-nucleotide indels. Furthermore, the loss of a functional MMR system caused the mutation spectra of these species to converge, implying that the MMR system itself contributes to species-specific mutation patterns. Bpsm and indel rates varied among genome regions, but do not explain the more rapid evolutionary rates of genes on chromosome 2, which likely result from weaker purifying selection. More generally, the very low mutation rates of Vibrio species correlate inversely with their immense population sizes and suggest that selection may not only have maximized replication fidelity but also optimized other polygenic traits relative to the constraints of genetic drift. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  17. Bacteriemia por Vibrio cholerae no-O1, no-O139 en un paciente en hemodiálisis crónica Non-O1, non-O139 Vibrio cholerae bacteremia in a chronic hemodialysis patient

    Directory of Open Access Journals (Sweden)

    Mariela S. Zárate

    2011-06-01

    Full Text Available Vibrio cholerae no-O1, no-O139 es un agente poco frecuente como causal de bacteriemias y no hay informes que documenten su presencia en pacientes en hemodiálisis crónica. Se describe el caso de una paciente en hemodiálisis crónica que presentó un cuadro de sepsis, por lo cual inició un tratamiento con vancomicina y ceftacidima. Al cabo de seis horas y media de incubación en el sistema BACT/ALERT de hemocultivo, se evidenció la presencia de bacilos curvos gram negativos, posteriormente identificados como Vibrio cholerae mediante pruebas bioquímicas convencionales y el uso de los kits API 20 NE y VITEK 2. La evaluación del serogrupo y de la presencia de factores de patogenicidad, realizada en el laboratorio de referencia, determinó que el microorganismo hallado pertenecía al serogrupo no-O1, no-O139. No se detectó la toxina de cólera, tampoco el factor de colonización ni la toxina termoestable. El aislamiento presentó sensibilidad frente a ampicilina, trimetoprima-sulfametoxazol, ciprofloxacina, tetraciclina, ceftacidima y cefotaxima por el método de difusión con discos y por VITEK 2. La paciente cumplió 14 días de tratamiento con ceftacidima endovenosa, con evolución favorable.Non-O1, and non-O139 Vibrio cholerae is an infrequent cause of bacteremia. There are no reports of such bacteremia in chronic hemodialysis patients. This work describes the case of a chronic hemodialysis patient that had an episode of septicemia associated with dialysis. Blood cultures were obtained and treatment was begun with vancomycin and ceftazidime. After 6.5 hours of incubation in the Bact/Alert system there is evidence of gram-negative curved bacilli that were identified as Vibrio cholerae by conventional biochemical tests, API 20 NE and the VITEK 2 system. This microorganism was sent to the reference laboratory for evaluation of serogroup and virulence factors and was identified as belonging to the non-O1 and non-O139 serogroup. The cholera

  18. Role of Indole Production on Virulence of Vibrio cholerae Using Galleria mellonella Larvae Model.

    Science.gov (United States)

    Nuidate, Taiyeebah; Tansila, Natta; Saengkerdsub, Suwat; Kongreung, Jetnaphang; Bakkiyaraj, Dhamodharan; Vuddhakul, Varaporn

    2016-09-01

    Cell to cell communication facilitated by chemical signals plays crucial roles in regulating various cellular functions in bacteria. Indole, one such signaling molecule has been demonstrated to control various bacterial phenotypes such as biofilm formation and virulence in diverse bacteria including Vibrio cholerae. The present study explores some key factors involved in indole production and the subsequent pathogenesis of V. cholerae. Indole production was higher at 37 °C than at 30 °C, although the growth at 37 °C was slightly higher. A positive correlation was observed between indole production and biofilm formation in V. cholerae. Maximum indole production was detected at pH 7. There was no significant difference in indole production between clinical and environmental V. cholerae isolates, although indole production in one environmental isolate was significantly different. Both growth and indole production showed relevant changes with differences in salinity. An indole negative mutant strain was constructed using transposon mutagenesis and the direct effect of indole on the virulence of V. cholerae was evaluated using Galleria mellonella larvae model. Comparison to the wild type strain, the mutant significantly reduced the mortality of G. mellonella larvae which regained its virulence after complementation with exogenous indole. A gene involved in indole production and the virulence of V. cholerae was identified.

  19. Virulence of an emerging pathogenic lineage of Vibrio nigripulchritudo is dependent on two plasmids.

    Science.gov (United States)

    Le Roux, Frédérique; Labreuche, Yannick; Davis, Brigid M; Iqbal, Naeem; Mangenot, Sophie; Goarant, Cyrille; Mazel, Didier; Waldor, Matthew K

    2011-02-01

    Vibrioses are the predominant bacterial infections in marine shrimp farms. Vibrio nigripulchritudo is an emerging pathogen of the cultured shrimp Litopenaeus stylirostris in New Caledonia and other regions in the Indo-Pacific. The molecular determinants of V. nigripulchritudo pathogenicity are unknown; however, molecular epidemiological studies have revealed that recent pathogenic V. nigripulchritudo isolates from New Caledonia all cluster into a monophyletic clade and contain a small plasmid, pB1067. Here, we report that a large plasmid, pA1066 (247 kb), can also serve as a marker for virulent V. nigripulchritudo, and that an ancestral version of this plasmid was likely acquired prior to other virulence-linked markers. Additionally, we demonstrate that pA1066 is critical for the full virulence of V. nigripulchritudo in several newly developed experimental models of infection. Plasmid pB1067 also contributes to virulence; only strains containing both plasmids induced the highest level of shrimp mortality. Thus, it appears that these plasmids, which are absent from non-pathogenic isolates, may be driving forces, as well as markers, for the emergence of a pathogenic lineage of V. nigripulchritudo.

  20. Toxigenic Vibrio cholerae identified in estuaries of Tanzania using PCR techniques.

    Science.gov (United States)

    Dalusi, Lucy; Lyimo, Thomas J; Lugomela, Charles; Hosea, Ken M M; Sjöling, Sara

    2015-03-01

    The current study assessed the occurrence of the Vibrio cholerae serogroups O1 and O139 in environmental samples along salinity gradients in three selected estuaries of Tanzania both through culture independent methods and by cultured bacteria. Occurrence of V. cholerae was determined by PCR targeting the V. cholerae outer membrane protein gene ompW. Furthermore, the presence of toxigenic strains and serogroups O1 and O139 was determined using multiplex PCR with specific primers targeting the cholera toxin gene subunit A, ctxA, and serotype specific primers, O1-rfb and O139-rfb, respectively. Results showed that V. cholerae occurred in approximately 10% (n = 185) of both the environmental samples and isolated bacteria. Eight of the bacteria isolates (n = 43) were confirmed as serogroup O1 while one belonged to serogroup O139, the first reported identification of this epidemic strain in East African coastal waters. All samples identified as serogroup O1 or O139 and a number of non-O1/O139 strains were ctxA positive. This study provides in situ evidence of the presence of pathogenic V. cholerae O1 and O139 and a number of V. cholerae non-O1/O139 that carry the cholera toxin gene in estuaries along the coast of Tanzania. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Integration of a laterally acquired gene into a cell network important for growth in a strain of Vibrio rotiferianus

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    Labbate Maurizio

    2011-11-01

    Full Text Available Abstract Background Lateral Gene Transfer (LGT is a major contributor to bacterial evolution and up to 25% of a bacterium's genome may have been acquired by this process over evolutionary periods of time. Successful LGT requires both the physical transfer of DNA and its successful incorporation into the host cell. One system that contributes to this latter step by site-specific recombination is the integron. Integrons are found in many diverse bacterial Genera and is a genetic system ubiquitous in vibrios that captures mobile DNA at a dedicated site. The presence of integron-associated genes, contained within units of mobile DNA called gene cassettes makes up a substantial component of the vibrio genome (1-3%. Little is known about the role of this system since the vast majority of genes in vibrio arrays are highly novel and functions cannot be ascribed. It is generally regarded that strain-specific mobile genes cannot be readily integrated into the cellular machinery since any perturbation of core metabolism is likely to result in a loss of fitness. Results In this study, at least one mobile gene contained within the Vibrio rotiferianus strain DAT722, but lacking close relatives elsewhere, is shown to greatly reduce host fitness when deleted and tested in growth assays. The precise role of the mobile gene product is unknown but impacts on the regulation of outermembrane porins. This demonstrates that strain specific laterally acquired mobile DNA can be integrated rapidly into bacterial networks such that it becomes advantageous for survival and adaptation in changing environments. Conclusions Mobile genes that are highly strain specific are generally believed to act in isolation. This is because perturbation of existing cell machinery by the acquisition of a new gene by LGT is highly likely to lower fitness. In contrast, we show here that at least one mobile gene, apparently unique to a strain, encodes a product that has integrated into central

  2. Vibrio Pathogens: A Public Health Concern in Rural Water Resources in Sub-Saharan Africa.

    Science.gov (United States)

    Osunla, Charles A; Okoh, Anthony I

    2017-10-07

    Members of the Vibrio genus are autochthonous inhabitants of aquatic environments and play vital roles in sustaining the aquatic milieu. The genus comprises about 100 species, which are mostly of marine or freshwater origin, and their classification is frequently updated due to the continuous discovery of novel species. The main route of transmission of Vibrio pathogens to man is through drinking of contaminated water and consumption inadequately cooked aquatic food products. In sub-Saharan Africa and much of the developing world, some rural dwellers use freshwater resources such as rivers for domestic activities, bathing, and cultural and religious purposes. This review describes the impact of inadequately treated sewage effluents on the receiving freshwater resources and the associated risk to the rural dwellers that depends on the water. Vibrio infections remain a threat to public health. In the last decade, Vibrio disease outbreaks have created alertness on the personal, economic, and public health uncertainties associated with the impact of contaminated water in the aquatic environment of sub-Saharan Africa. In this review, we carried out an overview of Vibrio pathogens in rural water resources in Sub-Saharan Africa and the implication of Vibrio pathogens on public health. Continuous monitoring of Vibrio pathogens among environmental freshwater and treated effluents is expected to help reduce the risk associated with the early detection of sources of infection, and also aid our understanding of the natural ecology and evolution of Vibrio pathogens.

  3. Vibrio Pathogens: A Public Health Concern in Rural Water Resources in Sub-Saharan Africa

    Directory of Open Access Journals (Sweden)

    Charles A. Osunla

    2017-10-01

    Full Text Available Members of the Vibrio genus are autochthonous inhabitants of aquatic environments and play vital roles in sustaining the aquatic milieu. The genus comprises about 100 species, which are mostly of marine or freshwater origin, and their classification is frequently updated due to the continuous discovery of novel species. The main route of transmission of Vibrio pathogens to man is through drinking of contaminated water and consumption inadequately cooked aquatic food products. In sub-Saharan Africa and much of the developing world, some rural dwellers use freshwater resources such as rivers for domestic activities, bathing, and cultural and religious purposes. This review describes the impact of inadequately treated sewage effluents on the receiving freshwater resources and the associated risk to the rural dwellers that depends on the water. Vibrio infections remain a threat to public health. In the last decade, Vibrio disease outbreaks have created alertness on the personal, economic, and public health uncertainties associated with the impact of contaminated water in the aquatic environment of sub-Saharan Africa. In this review, we carried out an overview of Vibrio pathogens in rural water resources in Sub-Saharan Africa and the implication of Vibrio pathogens on public health. Continuous monitoring of Vibrio pathogens among environmental freshwater and treated effluents is expected to help reduce the risk associated with the early detection of sources of infection, and also aid our understanding of the natural ecology and evolution of Vibrio pathogens.

  4. The Occurrence of Vibrio species in the Gut of Sardinella madrensis ...

    African Journals Online (AJOL)

    The occurrence of vibrio bacteria in the gut of “Songu”: Sardinella madrensis was investigated using enrichment procedures. Seventy percent (70%) of the total fish samples examined were positive for vibrios. The mean bacterial counts ranged between 2.68 x 102 to 1.30 x 104 cfu/g in all the fish samples. The weight of fish ...

  5. Bactericidal effect of lactoferrin and lactoferrin chimera against halophilic Vibrio parahaemolyticus

    NARCIS (Netherlands)

    Leon-Sicairos, N.; Canizalez-Roman, A.; de la Garza, M.; Reyes-Lopez, M.; Zazueta-Beltran, J.; Nazmi, K.; Gomez-Gil, B.; Bolscher, J.G.

    2009-01-01

    Infections caused by Vibrio parahaemolyticus, an halophilic member of the genus Vibrio, have increased globally in the last 5 years. Diarrhea caused by V. parahaemolyticus results from eating raw or undercooked seafood. The aim of this work was to investigate whether lactoferrin and some

  6. Efek Antibakteri Ekstrak Daun Mimba (Azadirachta indica A. Juss terhadap Bakteri Vibrio algynoliticus Secara In Vitro

    Directory of Open Access Journals (Sweden)

    Uli Ayini

    2014-03-01

    Full Text Available Budidaya udang windu di Indonesia telah berkembang pesat. Salah satu kendala budidaya udang adalah penyakit Vibriosis yang disebabkan oleh bakteri Vibrio algynoliticus. Tujuan penelitian ini adalah untuk mengetahui efek antibakeri ekstrak daun mimba terhadap bakteri Vibrio algynoliticus. Penelitian ini menggunakan metode dilusi untuk mengetahui efek antibakteri ekstrak daun mimba terhadap bakteri Vibrio algynoliticus secara in vitro. Konsentrasi ekstrak yang digunakan (% yaitu: 0; 2,5; 5; 7,5; 10; 12,5 dan sebagai kontrol terdiri dari kontrol positif, dan kontrol negatif. Pengumpulan data untuk menentukan MIC (Minimum Inhibitory Concentration dilakukan dengan membandingkan kejernihan kultur di medium TSB 2% pada berbagai konsentrasi yang berbeda, dengan kontrol positif dan kontrol negatif. Penentuan MBC (Minimum Bacterisidal Concentration dilakukan dengan melihat ada tidaknya dan jumlah koloni bakteri Vibrio alginolyticus yang muncul pada medium subkultur TSA 2% setelah inkubasi 24 jam. Hasil penelitian menunjukkan nilai MIC yaitu konsentrasi 5%, hal ini ditunjukkan dengan tabung yang mulai jernih. Nilai MBC ekstrak daun mimba terhadap bakteri Vibrio alginolyticus adalah konsentrasi 12,5% ditandai dengan sudah tidak munculnya  koloni bakteri Vibrio alginolyticus. Berdasarkan penelitian ini dapat disimpulkan bahwa ekstrak daun mimba dapat memberikan efek antibakteri terhadap bakteri Vibrio alginolyticus secara in vitro.Tiger shrimp cultivation in Indonesia has been growing rapidly. The main obstacle is the shrimp farming vibriosis disease caused by the bacterium Vibrio algynoliticus. The aim of this research was to determine the effects of neem leaf extract antibakeri against Vibrio algynoliticus. This study used a dilution method to determine the antibacterial effect of neem leaf extract against Vibrio algynoliticus bacteria in vitro. The concentration of the extract used (%: 0; 2.5; 5; 7.5; 10; 12.5 and as a control consisting of a positive

  7. Vibrio parahaemolyticus: A Review on the Pathogenesis, Prevalence and Advance Molecular Identification Techniques

    Directory of Open Access Journals (Sweden)

    Vengadesh eLetchumanan

    2014-12-01

    Full Text Available Vibrio parahaemolyticus is a Gram-negative halophilic bacterium that is found in estuarine, marine and coastal environments. Vibrio parahaemolyticus is the leading causal agent of human acute gastroenteritis following the consumption of raw, undercooked or mishandled marine products. In rare cases, Vibrio parahaemolyticus causes wound infection, ear infection or septicaemia in individuals with pre-existing medical conditions. Vibrio parahaemolyticus has two hemolysins virulence factors that are thermostable direct hemolysin (tdh-a pore-forming protein that contributes to the invasiveness of the bacterium in humans, and TDH-related hemolysin (trh, which plays a similar role as thermostable direct hemolysin (tdh in the disease pathogenesis. In addition, the bacterium is also encodes for adhesions and type III secretion systems (T3SS1 and T3SS2 to ensure its survival in the environment. This review aims at discussing the Vibrio parahemolyticus growth and characteristics, pathogenesis, prevalence and advances in molecular identification techniques.

  8. Vibrio ecology in PNW - The Ecology of Vibrio parahaemolyticus in the Pacific Northwest: Implications for risk assessment and early warning systems

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Over the past decade, there has been a significant increase in Vibrio parahaemolyticus-related gastroenteritis from the consumption of raw oysters harvested in...

  9. Saharan dust nutrients promote Vibrio bloom formation in marine surface waters

    Science.gov (United States)

    Westrich, Jason R.; Ebling, Alina M.; Landing, William M.; Joyner, Jessica L.; Kemp, Keri M.; Griffin, Dale W.; Lipp, Erin K.

    2016-01-01

    Vibrio is a ubiquitous genus of marine bacteria, typically comprising a small fraction of the total microbial community in surface waters, but capable of becoming a dominant taxon in response to poorly characterized factors. Iron (Fe), often restricted by limited bioavailability and low external supply, is an essential micronutrient that can limit Vibrio growth. Vibrio species have robust metabolic capabilities and an array of Fe-acquisition mechanisms, and are able to respond rapidly to nutrient influx, yet Vibrio response to environmental pulses of Fe remains uncharacterized. Here we examined the population growth of Vibrioafter natural and simulated pulses of atmospherically transported Saharan dust, an important and episodic source of Fe to tropical marine waters. As a model for opportunistic bacterial heterotrophs, we demonstrated that Vibrio proliferate in response to a broad range of dust-Fe additions at rapid timescales. Within 24 h of exposure, strains of Vibrio cholerae and Vibrio alginolyticus were able to directly use Saharan dust–Fe to support rapid growth. These findings were also confirmed with in situ field studies; arrival of Saharan dust in the Caribbean and subtropical Atlantic coincided with high levels of dissolved Fe, followed by up to a 30-fold increase of culturable Vibrio over background levels within 24 h. The relative abundance of Vibrio increased from ∼1 to ∼20% of the total microbial community. This study, to our knowledge, is the first to describe Vibrio response to Saharan dust nutrients, having implications at the intersection of marine ecology, Fe biogeochemistry, and both human and environmental health.

  10. Quorum Sensing Disruption in Vibrio harveyi Bacteria by Clay Materials.

    Science.gov (United States)

    Naik, Sajo P; Scholin, Jonathon; Ching, San; Chi, Fang; Herpfer, Marc

    2018-01-10

    This work describes the use of clay minerals as catalysts for the degradation of quorum sensing molecule N-(3-oxooctanoyl)-dl-homoserine lactone. Certain clay minerals as a result of their surface properties and porosity can catalytically degrade the quorum sensing molecule into smaller fragments. The disruption of quorum sensing by clay in a growing Gram-negative Vibrio harveyi bacteria culture was also studied by monitoring luminescence and population density of the bacteria, wherein quenching of bacterial quorum sensing activity was observed by means of luminescence reduction. The results of this study show that food-grade clays can be used as biocatalysts in disrupting bacterial activity in various media.

  11. Inhibitory Effect of Glycerin on Vibrio parahaemolyticus and Salmonella

    Science.gov (United States)

    Chun, Doki; Seol, Sung Yong; Tak, Ryunbin; Park, Cheong Kyu

    1972-01-01

    In a study of the effect of glycerin in transport media on Vibrio parahaemolyticus and Salmonella, it was found that a concentration of 30% glycerin was highly inhibitory for V. parahaemolyticus and to a lesser degree for Salmonella. The incorporation of peptone or human feces in media did not reduce the inhibitory effect of glycerin. In media with 15% glycerin, viable counts of V. parahaemolyticus and Salmonella increased after 24 hr of incubation both in the presence and absence of feces. Due to the concurrent increase in the total bacterial count in the media containing feces, no enrichment effect was noted. PMID:4565633

  12. Unsupervised statistical identification of genomic islands using ...

    Indian Academy of Sciences (India)

    Abstract. Vibrio cholerae, Vibrio vulnificus, Vibrio parahaemolyticus and sev- eral other related Vibrio species show distinctly similar two-chromosomal genome organization. However, the modes of pathogenicity are very different among these species, and this is largely attributed to externally acquired genetic elements.

  13. Insight Into the Origin and Evolution of the Vibrio parahaemolyticus Pandemic Strain

    Directory of Open Access Journals (Sweden)

    Romilio T. Espejo

    2017-07-01

    Full Text Available A strain of Vibrio parahaemolyticus that emerged in 1995 caused the first known pandemic involving this species. This strain comprises clonal autochthonous ocean-dwelling bacteria whose evolution has occurred in the ocean environment. The low sequence diversity in this population enabled the discovery of information on its origin and evolution that has been hidden in bacterial clones that have evolved over a long period. Multilocus sequencing and microarray analysis, together with phylogenetic analysis, of pandemic and pre-pandemic isolates has suggested that the founder clone was an O3:K6 non-pathogenic strain that initially acquired a toxRS/new region and subsequently acquired at least seven novel genomic islands. Sequencing and comparison of whole genomes later confirmed these early observations, and it confirmed that most of the genetic changes occurred via gene conversion involving horizontally transmitted DNA. The highly clonal population rapidly diversified, especially in terms of antigenicity, and 27 serotypes have already been reported. Comparisons of the core genomes derived from the founder clone indicate that there are only a few hundred single-nucleotide variations between isolates. However, when the whole genome is considered (the core plus non-core genome and from any clonal frame, the amount of DNA with a different clonal frame can reach up to 4.2% and the number of single-nucleotide variations can reach several hundred thousand. Altogether, these and previous observations based on multilocus sequence typing, microarray analysis, and whole-genome sequencing indicate the large contribution made by DNA with different clonal genealogy to genome diversification. The evidence also indicates that horizontal gene transfer (HGT caused the emergence of new pathogens. Furthermore, the extent of HGT seems to depend on the vicissitudes of the life of each bacterium, as exemplified by differences in thousands of base pairs acquired by HGT

  14. Application of Reverse Transcriptase-PCR-DGGE as a rapid method for routine determination of Vibrio spp. in foods.

    Science.gov (United States)

    Chahorm, Kanchana; Prakitchaiwattana, Cheunjit

    2018-01-02

    The aim of this research was to evaluate the feasibility of PCR-DGGE and Reverse Transcriptase-PCR-DGGE techniques for rapid detection of Vibrio species in foods. Primers GC567F and 680R were initially evaluated for amplifying DNA and cDNA of ten references Vibrio species by PCR method. The GC-clamp PCR amplicons were separated according to their sequences by the DGGE using 10% (w/v) polyacrylamide gel containing 45-70% urea and formamide denaturants. Two pair of Vibrio species, which could not be differentiated on the gel, was Vibrio fluvialis - Vibrio furnissii and Vibrio parahaemolyticus - Vibrio harveyi. To determine the detection limit, in the community of 10 reference strains containing the same viable population, distinct DNA bands of 3 species; Vibrio cholerae, Vibrio mimicus and Vibrio alginolyticus were consistently observed by PCR-DGGE technique. In fact, 5 species; Vibrio cholerae, Vibrio mimicus, Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio fluvialis consistently observed by Reverse Transcriptase-PCR-DGGE. In the community containing different viable population increasing from 10 2 to 10 5 CFU/mL, PCR-DGGE analysis only detected the two most prevalent species, while RT-PCR-DGGE detected the five most prevalent species. Therefore, Reverse Transcriptase-PCR-DGGE was also selected for detection of various Vibrio cell conditions, including viable cell (VC), injured cells from frozen cultures (IVC) and injured cells from frozen cultures with pre-enrichment (PIVC). It was found that cDNA band of all cell conditions gave the same migratory patterns, except that multiple cDNA bands of Plesiomonas shigelloides under IVC and PIVC conditions were found. When Reverse Transcriptase-PCR-DGGE was used for detecting Vibrio parahaemolyticus in the pathogen-spiked food samples, Vibrio parahaemolyticus could be detected in the spiked samples containing at least 10 2 CFU/g of this pathogen. The results obtained also corresponded to standard method (USFDA, 2004

  15. Vitroprocines, new antibiotics against Acinetobacter baumannii, discovered from marine Vibrio sp. QWI-06 using mass-spectrometry-based metabolomics approach

    Science.gov (United States)

    Liaw, Chih-Chuang; Chen, Pei-Chin; Shih, Chao-Jen; Tseng, Sung-Pin; Lai, Ying-Mi; Hsu, Chi-Hsin; Dorrestein, Pieter C.; Yang, Yu-Liang

    2015-08-01

    A robust and convenient research strategy integrating state-of-the-art analytical techniques is needed to efficiently discover novel compounds from marine microbial resources. In this study, we identified a series of amino-polyketide derivatives, vitroprocines A-J, from the marine bacterium Vibrio sp. QWI-06 by an integrated approach using imaging mass spectroscopy and molecular networking, as well as conventional bioactivity-guided fractionation and isolation. The structure-activity relationship of vitroprocines against Acinetobacter baumannii is proposed. In addition, feeding experiments with 13C-labeled precursors indicated that a pyridoxal 5‧-phosphate-dependent mechanism is involved in the biosynthesis of vitroprocines. Elucidation of amino-polyketide derivatives from a species of marine bacteria for the first time demonstrates the potential of this integrated metabolomics approach to uncover marine bacterial biodiversity.

  16. Migratory responses of hemocytes to Vibrio parahaemolyticus in the alimentary tract of an estuarine neritid gastropod, Clithon retropictus.

    Science.gov (United States)

    Kumazawa, N H; Mine, A

    2001-12-01

    Migratory responses of hemocytes to Vibrio parahaemolyticus strain D3 in the alimentary tracts of an estuarine neritid gastropod, Clithon retropictus, and a related marine neritid, Nerita albicilla, were examined under the scanning electron microscope. After ingesting the strain, active responses were seen at the esophagus, stomach and anterior intestine of adult C. retropictus and at the middle and posterior intestines of adult N. albicilla. When the alimentary tracts were isolated from the gastropod and incubated in vitro with strain D3, active response was induced at the most parts of the tract of the adult gastropods and at the stomach and the anterior intestine of juvenile C. retropictus. The responding hemocytes were confirmed to be granulocytes in the semi-thin sections of the tract of adult C. retropictus. The poor hemocyte responses at the middle and posterior intestines of juvenile C. retropictus might support the colonization of the organism there.

  17. New insights into Oculina patagonica coral diseases and their associated Vibrio spp. communities.

    Science.gov (United States)

    Rubio-Portillo, Esther; Yarza, Pablo; Peñalver, Cindy; Ramos-Esplá, Alfonso A; Antón, Josefa

    2014-09-01

    Bleaching of Oculina patagonica has been extensively studied in the Eastern Mediterranean Sea, although no studies have been carried out in the Western basin. In 1996 Vibrio mediterranei was reported as the causative agent of bleaching in O. patagonica but it has not been related to bleached or healthy corals since 2003, suggesting that it was no longer involved in bleaching of O. patagonica. In an attempt to clarify the relationship between Vibrio spp., seawater temperature and coral diseases, as well as to investigate the putative differences between Eastern and Western Mediterranean basins, we have analysed the seasonal patterns of the culturable Vibrio spp. assemblages associated with healthy and diseased O. patagonica colonies. Two sampling points located in the Spanish Mediterranean coast were chosen for this study: Alicante Harbour and the Marine Reserve of Tabarca. A complex and dynamic assemblage of Vibrio spp. was present in O. patagonica along the whole year and under different environmental conditions and coral health status. While some Vibrio spp. were detected all year around in corals, the known pathogens V. mediteranei and V. coralliilyticus were only present in diseased specimens. The pathogenic potential of these bacteria was studied by experimental infection under laboratory conditions. Both vibrios caused diseased signs from 24 °C, being higher and faster at 28 °C. Unexpectedly, the co-inoculation of these two Vibrio species seemed to have a synergistic pathogenic effect over O. patagonica, as disease signs were readily observed at temperatures at which bleaching is not normally observed.

  18. Identification and Initial Characterization of Prophages in Vibrio campbellii.

    Directory of Open Access Journals (Sweden)

    Nicola Lorenz

    Full Text Available Phages are bacteria targeting viruses and represent the most abundant biological entities on earth. Marine environments are exceptionally rich in bacteriophages, harboring a total of 4x1030 viruses. Nevertheless, marine phages remain poorly characterized. Here we describe the identification of intact prophage sequences in the genome of the marine γ-proteobacterium Vibrio campbellii ATCC BAA-1116 (formerly known as V. harveyi ATCC BAA-1116, which presumably belong to the family of Myoviridae. One prophage was found on chromosome I and shows significant similarities to the previously identified phage ΦHAP-1. The second prophage region is located on chromosome II and is related to Vibrio phage kappa. Exposure of V. campbellii to mitomycin C induced the lytic cycle of two morphologically distinct phages and, as expected, extracellular DNA from induced cultures was found to be specifically enriched for the sequences previously identified as prophage regions. Heat stress (50°C, 30 min was also found to induce phage release in V. campbellii. Notably, promoter activity of two representative phage genes indicated heterogeneous phage induction within the population.

  19. Vibrio cholerae Represses Polysaccharide Synthesis To Promote Motility in Mucosa

    Science.gov (United States)

    Liu, Zhenyu; Wang, Yuning; Liu, Shengyan; Sheng, Ying; Rueggeberg, Karl-Gustav; Wang, Hui; Li, Jie; Gu, Frank X.; Zhong, Zengtao; Kan, Biao

    2015-01-01

    The viscoelastic mucus layer of gastrointestinal tracts is a host defense barrier that a successful enteric pathogen, such as Vibrio cholerae, must circumvent. V. cholerae, the causative agent of cholera, is able to penetrate the mucosa and colonize the epithelial surface of the small intestine. In this study, we found that mucin, the major component of mucus, promoted V. cholerae movement on semisolid medium and in liquid medium. A genome-wide screen revealed that Vibrio polysaccharide (VPS) production was inversely correlated with mucin-enhanced motility. Mucin adhesion assays indicated that VPS bound to mucin. Moreover, we found that vps expression was reduced upon exposure to mucin. In an infant mouse colonization model, mutants that overexpressed VPS colonized less effectively than wild-type strains in more distal intestinal regions. These results suggest that V. cholerae is able to sense mucosal signals and modulate vps expression accordingly so as to promote fast motion in mucus, thus allowing for rapid spread throughout the intestines. PMID:25561707

  20. Invariant recognition of polychromatic images of Vibrio cholerae 01

    Science.gov (United States)

    Alvarez-Borrego, Josue; Mourino-Perez, Rosa R.; Cristobal, Gabriel; Pech-Pacheco, Jose L.

    2002-04-01

    Cholera is an acute intestinal infectious disease. It has claimed many lives throughout history, and it continues to be a global health threat. Cholera is considered one of the most important emergence diseases due its relation with global climate changes. Automated methods such as optical systems represent a new trend to make more accurate measurements of the presence and quantity of this microorganism in its natural environment. Automatic systems eliminate observer bias and reduce the analysis time. We evaluate the utility of coherent optical systems with invariant correlation for the recognition of Vibrio cholerae O1. Images of scenes are recorded with a CCD camera and decomposed in three RGB channels. A numeric simulation is developed to identify the bacteria in the different samples through an invariant correlation technique. There is no variation when we repeat the correlation and the variation between images correlation is minimum. The position-, scale-, and rotation-invariant recognition is made with a scale transform through the Mellin transform. The algorithm to recognize Vibrio cholerae O1 is the presence of correlation peaks in the green channel output and their absence in red and blue channels. The discrimination criterion is the presence of correlation peaks in red, green, and blue channels.

  1. Vibrio fischeri-derived outer membrane vesicles trigger host development.

    Science.gov (United States)

    Aschtgen, Marie-Stephanie; Wetzel, Keith; Goldman, William; McFall-Ngai, Margaret; Ruby, Edward

    2016-04-01

    Outer membrane vesicles (OMV) are critical elements in many host-cell/microbe interactions. Previous studies of the symbiotic association between Euprymna scolopes and Vibrio fischeri had shown that within 12 h of colonizing crypts deep within the squid's light organ, the symbionts trigger an irreversible programme of tissue development in the host. Here, we report that OMV produced by V. fischeri are powerful contributors to this process. The first detectable host response to the OMV is an increased trafficking of macrophage-like cells called haemocytes into surface epithelial tissues. We showed that exposing the squid to other Vibrio species fails to induce this trafficking; however, addition of a high concentration of their OMV, which can diffuse into the crypts, does. We also provide evidence that tracheal cytotoxin released by the symbionts, which can induce haemocyte trafficking, is not part of the OMV cargo, suggesting two distinct mechanisms to induce the same morphogenesis event. By manipulating the timing and localization of OMV signal delivery, we showed that haemocyte trafficking is fully induced only when V. fischeri, the sole species able to reach and grow in the crypts, succeeds in establishing a sustained colonization. Further, our data suggest that the host's detection of OMV serves as a symbiotic checkpoint prior to inducing irreversible morphogenesis. © 2015 John Wiley & Sons Ltd.

  2. "Quorum Non-Sensing": Social Cheating and Deception in Vibrio cholerae.

    Science.gov (United States)

    Katzianer, David S; Wang, Hui; Carey, Ryan M; Zhu, Jun

    2015-06-01

    Quorum sensing (QS) is widely used by bacteria to coordinate behavior in response to external stimuli. In Vibrio cholerae, this process is important for environmental survival and pathogenesis, though, intriguingly, a large percentage of natural isolates are QS deficient. Here, we show that QS-deficient mutants can spread as social cheaters by ceasing production of extracellular proteases under conditions requiring their growth. We further show that mutants stimulate biofilm formation and are over-represented in biofilms compared to planktonic communities; on this basis, we suggest that QS-deficient mutants may have the side effect of enhancing environmental tolerance of natural populations due to the inherent resistance properties of biofilms. Interestingly, high frequencies of QS-deficient individuals did not impact production of QS signaling molecules despite mutants being unable to respond to these inducers, indicating that these variants actively cheat by false signaling under conditions requiring QS. Taken together, our results suggest that social cheating may drive QS deficiency emergence within V. cholerae natural populations. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  3. Visualization of coral host-pathogen interactions using a stable GFP-labeled Vibrio coralliilyticus strain

    Science.gov (United States)

    Pollock, F. Joseph; Krediet, Cory J.; Garren, Melissa; Stocker, Roman; Winn, Karina; Wilson, Bryan; Huete-Stauffer, Carla; Willis, Bette L.; Bourne, David G.

    2015-06-01

    The bacterium Vibrio coralliilyticus has been implicated as the causative agent of coral tissue loss diseases (collectively known as white syndromes) at sites across the Indo-Pacific and represents an emerging model pathogen for understanding the mechanisms linking bacterial infection and coral disease. In this study, we used a mini-Tn7 transposon delivery system to chromosomally label a strain of V. coralliilyticus isolated from a white syndrome disease lesion with a green fluorescent protein gene (GFP). We then tested the utility of this modified strain as a research tool for studies of coral host-pathogen interactions. A suite of biochemical assays and experimental infection trials in a range of model organisms confirmed that insertion of the GFP gene did not interfere with the labeled strain's virulence. Using epifluorescence video microscopy, the GFP-labeled strain could be reliably distinguished from non-labeled bacteria present in the coral holobiont, and the pathogen's interactions with the coral host could be visualized in real time. This study demonstrates that chromosomal GFP labeling is a useful technique for visualization and tracking of coral pathogens and provides a novel tool to investigate the role of V. coralliilyticus in coral disease pathogenesis.

  4. The putative oligosaccharide translocase SypK connects biofilm formation with quorum signaling in Vibrio fischeri.

    Science.gov (United States)

    Miyashiro, Tim; Oehlert, Dane; Ray, Valerie A; Visick, Karen L; Ruby, Edward G

    2014-12-01

    Quorum signaling (QS) describes how bacteria can use small signaling molecules (autoinducers) to coordinate group-level behaviors. In Vibrio fischeri, QS is achieved through a complex regulatory network that ultimately controls bioluminescence, motility, and host colonization. We conducted a genetic screen focused on qrr1, which encodes a small regulatory RNA that is necessary for the core quorum-signaling cascade to transduce autoinducer information into cellular responses. We isolated unique mutants with a transposon inserted into one of two genes within the syp locus, which is involved in biofilm formation. We found that overexpression of sypK, which encodes a putative oligosaccharide translocase, is sufficient to activate qrr1, and, in addition, this effect appears to depend on the kinase activity of the sensor LuxQ. Consistent with the established model for QS in V. fischeri, enhanced expression of qrr1 by the overexpression of sypK resulted in reduced bioluminescence and increased motility. Finally, we found that induction of the syp locus by overexpression of sypG was sufficient to activate qrr1 levels. Together, our results show how conditions that promote biofilm formation impact the quorum-signaling network in V. fischeri, and further highlight the integrated nature of the regulatory circuits involved in complex bacterial behaviors. © 2014 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  5. Vibrio fischeri exhibit the growth advantage in stationary-phase phenotype.

    Science.gov (United States)

    Petrun, Branden; Lostroh, C Phoebe

    2013-02-01

    Vibrio fischeri are bioluminescent marine bacteria that can be isolated from their symbiotic animal partners or from ocean water. A V. fischeri population increases exponentially inside the light organ of the Hawaiian bobtail squid (Euprymna scolopes) while the host is quiescent during the day. This bacterial light organ population reaches stationary phase and then remains high during the night, when the squid use bacterial bioluminescence as a counter-predation strategy. At dawn, host squid release 90%-95% of the light organ contents into the ocean water prior to burying in the sand for the day. As the squid sleeps, the cycle of bacterial population growth in the light organ begins again. These V. fischeri cells that are vented into the ocean must persist under typical marine low nutrient conditions until they encounter another opportunity to colonize a host. We hypothesized that because V. fischeri regularly encounter cycles of feast and famine in nature, they would exhibit the growth advantage in stationary phase (GASP) phenotype. We found that older V. fischeri cells exhibit a Class 2 GASP response in which old cells increase dramatically in frequency while the population of young V. fischeri cells remains almost constant during co-incubation.

  6. A highly specific phage defense system is a conserved feature of the Vibrio cholerae mobilome.

    Directory of Open Access Journals (Sweden)

    Brendan J O'Hara

    2017-06-01

    Full Text Available Vibrio cholerae-specific bacteriophages are common features of the microbial community during cholera infection in humans. Phages impose strong selective pressure that favors the expansion of phage-resistant strains over their vulnerable counterparts. The mechanisms allowing virulent V. cholerae strains to defend against the ubiquitous threat of predatory phages have not been established. Here, we show that V. cholerae PLEs (phage-inducible chromosomal island-like elements are widespread genomic islands dedicated to phage defense. Analysis of V. cholerae isolates spanning a 60-year collection period identified five unique PLEs. Remarkably, we found that all PLEs (regardless of geographic or temporal origin respond to infection by a myovirus called ICP1, the most prominent V. cholerae phage found in cholera patient stool samples from Bangladesh. We found that PLE activity reduces phage genome replication and accelerates cell lysis following ICP1 infection, killing infected host cells and preventing the production of progeny phage. PLEs are mobilized by ICP1 infection and can spread to neighboring cells such that protection from phage predation can be horizontally acquired. Our results reveal that PLEs are a persistent feature of the V. cholerae mobilome that are adapted to providing protection from a single predatory phage and advance our understanding of how phages influence pathogen evolution.

  7. A highly specific phage defense system is a conserved feature of the Vibrio cholerae mobilome

    Science.gov (United States)

    O’Hara, Brendan J.

    2017-01-01

    Vibrio cholerae-specific bacteriophages are common features of the microbial community during cholera infection in humans. Phages impose strong selective pressure that favors the expansion of phage-resistant strains over their vulnerable counterparts. The mechanisms allowing virulent V. cholerae strains to defend against the ubiquitous threat of predatory phages have not been established. Here, we show that V. cholerae PLEs (phage-inducible chromosomal island-like elements) are widespread genomic islands dedicated to phage defense. Analysis of V. cholerae isolates spanning a 60-year collection period identified five unique PLEs. Remarkably, we found that all PLEs (regardless of geographic or temporal origin) respond to infection by a myovirus called ICP1, the most prominent V. cholerae phage found in cholera patient stool samples from Bangladesh. We found that PLE activity reduces phage genome replication and accelerates cell lysis following ICP1 infection, killing infected host cells and preventing the production of progeny phage. PLEs are mobilized by ICP1 infection and can spread to neighboring cells such that protection from phage predation can be horizontally acquired. Our results reveal that PLEs are a persistent feature of the V. cholerae mobilome that are adapted to providing protection from a single predatory phage and advance our understanding of how phages influence pathogen evolution. PMID:28594826

  8. Flavonoids from Piper delineatum modulate quorum-sensing-regulated phenotypes in Vibrio harveyi.

    Science.gov (United States)

    Martín-Rodríguez, Alberto J; Ticona, Juan C; Jiménez, Ignacio A; Flores, Ninoska; Fernández, José J; Bazzocchi, Isabel L

    2015-09-01

    Quorum sensing (QS), or bacterial cell-to-cell communication, is a key process for bacterial colonization of substrata through biofilm formation, infections, and production of virulence factors. In an ongoing investigation of bioactive secondary metabolites from Piper species, four new flavonoids (1-4), along with five known ones (5-9) were isolated from the leaves of Piper delineatum. Their stereostructures were established by spectroscopic and spectrometric methods, including 1D and 2D NMR experiments, and comparison with data reported in the literature. The compounds were screened for their ability to interfere with QS signaling in the bacterial model Vibrio harveyi. Four compounds from this series (2, 3, 6, and 7) exhibited remarkable activity in the micromolar range, being compounds 3 and 7 particularly attractive since they did not affect bacterial growth. The results suggest that these flavonoids disrupt QS-mediated bioluminescence by interaction with elements downstream LuxO in the QS circuit of V. harveyi, and also, they exhibited a strong dose-dependent inhibition of biofilm formation. The present findings shed light on the QS inhibition mechanisms of flavonoids, underlining their potential applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Characterization of Aeromonas trota strains that cross-react with Vibrio cholerae O139 Bengal.

    Science.gov (United States)

    Albert, M J; Ansaruzzaman, M; Shimada, T; Rahman, A; Bhuiyan, N A; Nahar, S; Qadri, F; Islam, M S

    1995-12-01

    It has previously been shown that Vibrio cholerae O139 Bengal shares antigens with V. cholerae serogroups O22 and O155. We detected six surface water isolates of Aeromonas trota that agglutinated in polyclonal antisera to V. cholerae O139 and V. cholerae O22 but not in antiserum to V. cholerae O155. On the basis of agglutinin-absorption studies, the antigenic relationship between the cross-reacting bacteria were found to be in an a,b-a,c fashion, where a is the common antigenic epitope and b and c are unique epitopes. The antigen sharing between A. trota strains and V. cholerae O139 was confirmed in immunoblot studies. However, A. trota strains did not react with two monoclonal antibodies specific for V. cholerae O139 and, consequently, tested negative in the Bengal SMART rapid diagnostic test for V. cholerae O139 which uses one of the monoclonal antibodies. A polyclonal antiserum to a cross-reacting A. trota strain cross-protected infant mice against cholera on challenge with virulent V. cholerae O139. All A. trota strains were cytotoxic for HeLa cells, positive for adherence to HEp-2 cells, and weakly invasive for HEp-2 cells; one strain was heat-stable toxin positive in the suckling mouse assay; however, all strains were negative for cholera toxin-like enterotoxin. Studies on bacteria that share somatic antigen with V. cholerae O139 may shed further light on the genesis of V. cholerae O139.

  10. Susceptibility of Vibrio cholerae O139 to Antibody-Dependent, Complement-Mediated Bacteriolysis

    Science.gov (United States)

    Attridge, Stephen R.; Qadri, Firdausi; Albert, M. John; Manning, Paul A.

    2000-01-01

    Volunteer studies with Vibrio cholerae O1 have shown that the best correlate of a vaccine's protective efficacy is its propensity to elicit serum bactericidal responses in its recipients. Attempts to detect such responses following infection with V. cholerae O139, however, have met with varying success. Using a tube-based assay which involves viable counting, we now report that strains of serogroup O139 can appear to be sensitive or resistant to a fixed concentration of complement in the presence of antibody, depending on assay conditions. Susceptibility to lysis is critically dependent on the availability of complement, but with O139 indicator strains this is not simply determined by the concentration of serum added to the reaction mix. The nature of the assay diluent and the concentration of indicator bacteria can also dramatically affect bactericidal end points, whereas such variables have minimal significance with O1 indicator bacteria. Although some laboratories use unencapsulated mutant strains to seek evidence of seroconversion following exposure to V. cholerae O139, this is not necessary, and our findings question the significance of capsule expression as a determinant of complement sensitivity when antibody is present. The medium used for growth of the indicator strain and the particular strain used appeared to be unimportant. Each of seven O139 isolates tested was found to be lysed by antibody and complement in our standard assay system, which allowed the detection of significant serum bactericidal responses in 9 of 11 cases of O139 disease. PMID:10799459

  11. Intraspecific Competition Impacts Vibrio fischeri Strain Diversity during Initial Colonization of the Squid Light Organ.

    Science.gov (United States)

    Sun, Yan; LaSota, Elijah D; Cecere, Andrew G; LaPenna, Kyle B; Larios-Valencia, Jessie; Wollenberg, Michael S; Miyashiro, Tim

    2016-05-15

    Animal development and physiology depend on beneficial interactions with microbial symbionts. In many cases, the microbial symbionts are horizontally transmitted among hosts, thereby making the acquisition of these microbes from the environment an important event within the life history of each host. The light organ symbiosis established between the Hawaiian squid Euprymna scolopes and the bioluminescent bacterium Vibrio fischeri is a model system for examining how hosts acquire horizontally transmitted microbial symbionts. Recent studies have revealed that the light organ of wild-caught E. scolopes squid contains polyclonal populations of V. fischeri bacteria; however, the function and development of such strain diversity in the symbiosis are unknown. Here, we report our phenotypic and phylogenetic characterizations of FQ-A001, which is a V. fischeri strain isolated directly from the light organ of an E. scolopes individual. Relative to the type strain ES114, FQ-A001 exhibits similar growth in rich medium but displays increased bioluminescence and decreased motility in soft agar. FQ-A001 outcompetes ES114 in colonizing the crypt spaces of the light organs. Remarkably, we find that animals cocolonized with FQ-A001 and ES114 harbor singly colonized crypts, in contrast to the cocolonized crypts observed from competition experiments involving single genotypes. The results with our two-strain system suggest that strain diversity within the squid light organ is a consequence of diversity in the single-strain colonization of individual crypt spaces. The developmental programs and overall physiologies of most animals depend on diverse microbial symbionts that are acquired from the environment. However, the basic principles underlying how microbes colonize their hosts remain poorly understood. Here, we report our findings of bacterial strain competition within the coevolved animal-microbe symbiosis composed of the Hawaiian squid and bioluminescent bacterium Vibrio fischeri

  12. Ecología de Vibrio cholerae en relación al Fitoplancton y variables fisicoquímicas en ríos de Tucumán (Argentina Ecology of Vibrio cholerae in relation to phytoplankton and physico-chemical variables in rivers of Tucumán (Argentina

    Directory of Open Access Journals (Sweden)

    V. Mirande

    relation between phytoplankton species, physico-chemical variables and V. cholerae isolations in rivers of Tucumán. There were 18 sampling in the rivers Salí and Lules from 2003 to 2005. Physico-chemical variables of water (pH, temperature, conductivity and dissolved oxygen, phytoplankton (richness and relative frequency and V. cholerae isolations were studied in this paper. Results showed differences in water quality, with anoxic periods in the River Salí. Diatoms were conspicuous in almost all months and generally there were in percentages upper 85 %. Just Vibrio cholerae non-O1, non-O139 was isolated, specially in months warmer with alkaline pH and also with dissolved oxygen low concentration.

  13. Diluent composition for use of API 20E in characterizing marine and estuarine bacteria.

    Science.gov (United States)

    MacDonell, M T; Singleton, F L; Hood, M A

    1982-01-01

    Nine chemically defined inoculation diluents, with compositions ranging from 0.85% NaCl to 35% marine salts, were used to evaluate the influence of diluent composition on the biochemical profiles of 30 marine and estuarine bacterial strains, including species of Vibrio, Aeromonas, Allomonas, and Photobacterium. Results demonstrated that a 20% marine salts diluent enabled the characterization of halophilic strains normally nonreactive by the API 20E system. Furthermore, the use of 20% marine salts showed that certain environmental isolates, identifiable as Vibrio parahaemolyticus by the recommended clinical inoculation procedure, were Vibrio vulnificus. An analysis of the profiles provided by the nine diluents indicates that the API 20E system, modified by the use of a diluent composed of 20% marine salts and incubated at 22 degrees C, can provide a reliable tool for the rapid characterization of marine and estuarine bacterial isolates. PMID:7125655

  14. Diluent composition for use of API 20E in characterizing marine and estuarine bacteria.

    Science.gov (United States)

    MacDonell, M T; Singleton, F L; Hood, M A

    1982-08-01

    Nine chemically defined inoculation diluents, with compositions ranging from 0.85% NaCl to 35% marine salts, were used to evaluate the influence of diluent composition on the biochemical profiles of 30 marine and estuarine bacterial strains, including species of Vibrio, Aeromonas, Allomonas, and Photobacterium. Results demonstrated that a 20% marine salts diluent enabled the characterization of halophilic strains normally nonreactive by the API 20E system. Furthermore, the use of 20% marine salts showed that certain environmental isolates, identifiable as Vibrio parahaemolyticus by the recommended clinical inoculation procedure, were Vibrio vulnificus. An analysis of the profiles provided by the nine diluents indicates that the API 20E system, modified by the use of a diluent composed of 20% marine salts and incubated at 22 degrees C, can provide a reliable tool for the rapid characterization of marine and estuarine bacterial isolates.

  15. Salinity-induced survival strategy of Vibrio cholerae associated with copepods in Cochin backwaters

    Digital Repository Service at National Institute of Oceanography (India)

    Thomas, K.U.; Joseph, N.; Raveendran, O.; Nair, S.

    The occurrence of Vibrio cholerae in water, sediment and copepods was studied over a wide range of salinity using conventional and polymerase chain reaction (PCR) techniques in the Cochin backwaters. V. cholerae occurred either as culturable or non-culturable...

  16. Competitive Survival of Escherichia coli, Vibrio cholerae, Salmonella typhimurium and Shigella dysenteriae in Riverbed Sediments

    CSIR Research Space (South Africa)

    Abia, AL

    2016-11-01

    Full Text Available investigated the survival of Escherichia coli, Salmonella enterica ser. Typhimurium, Vibrio cholerae and Shigella dysenteriae in riverbed sediments of the Apies River. Experiments were performed in flow chambers containing three sediment types and connected...

  17. Corrosion of mild steel and stainless steel by marine Vibrio sp.

    Digital Repository Service at National Institute of Oceanography (India)

    PrabhaDevi; Wagh, A.B.

    Microbially induced corrosion (MIC) of stainless steel and mild steel coupons exposed to media with and without a bacterial culture Vibrio sp. was studied using Scanning Electron Microscope (SEM). Pitting type of corrosion was noticed which was more...

  18. Vibrio cholerae Detection in Water and Wastewater by Polymerase Chain Reaction Assay

    Directory of Open Access Journals (Sweden)

    Behnaz Barzamini

    2014-11-01

    Full Text Available Background: Vibrio cholerae is a significant human pathogen worldwide and annually causes some cases of deaths. Contaminated water plays an important role in transmission of this pathogen, which indicates the importance of early diagnosis. Objectives: The current study aimed to perform Polymerase Chain Reaction (PCR on water and wastewater samples to determine the detection limit for Vibrio cholerae. Materials and Methods: PCR was performed on the DNA extracted from Vibrio cholerae of the contaminated water and wastewater using ctxA gene specific primers. The accuracy of PCR method to detect these bacteria was also assessed. Results: The result of PCR performed on the extracted DNA showed a specific 241 base pair band. The limit of bacterial detection for water and wastewater were 40 cfu/mL and 81 cfu/mL, respectively. Conclusions: In the current study, PCR performance using the ctxA gene specific primers to detect Vibrio cholerae was found highly accurate and specific.

  19. Complete Genome Sequence of Vibrio fischeri: A Symbiotic Bacterium with Pathogenic Congeners

    National Research Council Canada - National Science Library

    E. G. Ruby; M. Urbanowski; J. Campbell; A. Dunn; M. Faini; R. Gunsalus; P. Lostroh; C. Lupp; J. McCann; D. Millikan; A. Schaefer; E. Stabb; A. Stevens; K. Visick; C. Whistler; E. P. Greenberg

    2005-01-01

    Vibrio fischeri belongs to the Vibrionaceae, a large family of marine γ-proteobacteria that includes several dozen species known to engage in a diversity of beneficial or pathogenic interactions with animal tissue...

  20. Global discovery of colonization determinants in the squid symbiont Vibrio fischeri

    National Research Council Canada - National Science Library

    John F. Brooks; Mattias C. Gyllborg; David C. Cronin; Sarah J. Quillin; Celeste A. Mallama; Randi Foxall; Cheryl Whistler; Andrew L. Goodman; Mark J. Mandel

    2014-01-01

    .... To understand the molecular determinants of microbiota selection, we examined colonization of a simplified model in which the light organ of Euprymna scolopes squid is colonized exclusively by Vibrio fischeri bacteria...

  1. Population structure of clinical Vibrio parahaemolyticus from 17 coastal countries, determined through multilocus sequence analysis.

    Science.gov (United States)

    Han, Dongsheng; Tang, Hui; Lu, Jun; Wang, Guangzhou; Zhou, Lin; Min, Lingfeng; Han, Chongxu

    2014-01-01

    Vibrio parahaemolyticus is a leading cause of food-borne gastroenteritis worldwide. Although this bacterium has been the subject of much research, the population structure of clinical strains from worldwide collections remains largely undescribed, and the recorded outbreaks of V. parahaemolyticus gastroenteritis highlight the need for the subtyping of this species. We present a broad phylogenetic analysis of 490 clinical V. parahaemolyticus isolates from 17 coastal countries through multilocus sequence analysis (MLST). The 490 tested isolates fell into 161 sequence types (STs). The eBURST algorithm revealed that the 161 clinically relevant STs belonged to 8 clonal complexes, 11 doublets, and 94 singletons, showing a high level of genetic diversity. CC3 was found to be a global epidemic clone of V. parahaemolyticus, and ST-3 was the only ST with an international distribution. recA was observed to be evolving more rapidly, exhibiting the highest degree of nucleotide diversity (0.028) and the largest number of polymorphic nucleotide sites (177). We also found that the high variability of recA was an important cause of differences between the results of the eBURST and ME tree analyses, suggesting that recA has a much greater influence on the apparent evolutionary classification of V. parahaemolyticus based on the current MLST scheme. In conclusion, it is evident that a high degree of genetic diversity within the V. parahaemolyticus population and multiple sequence ty