WorldWideScience

Sample records for vibrio harveyi electronic

  1. Isolation of lytic bacteriophage against Vibrio harveyi.

    Science.gov (United States)

    Crothers-Stomps, C; Høj, L; Bourne, D G; Hall, M R; Owens, L

    2010-05-01

    The isolation of lytic bacteriophage of Vibrio harveyi with potential for phage therapy of bacterial pathogens of phyllosoma larvae from the tropical rock lobster Panulirus ornatus. Water samples from discharge channels and grow-out ponds of a prawn farm in northeastern Australia were enriched for 24 h in a broth containing four V. harveyi strains. The bacteriophage-enriched filtrates were spotted onto bacterial lawns demonstrating that the bacteriophage host range for the samples included strains of V. harveyi, Vibrio campbellii, Vibrio rotiferianus, Vibrio parahaemolyticus and Vibrio proteolyticus. Bacteriophage were isolated from eight enriched samples through triple plaque purification. The host range of purified phage included V. harveyi, V. campbellii, V. rotiferianus and V. parahaemolyticus. Transmission electron microscope examination revealed that six purified phage belonged to the family Siphoviridae, whilst two belonged to the family Myoviridae. The Myoviridae appeared to induce bacteriocin production in a limited number of host bacterial strains, suggesting that they were lysogenic rather than lytic. A purified Siphoviridae phage could delay the entry of a broth culture of V. harveyi strain 12 into exponential growth, but could not prevent the overall growth of the bacterial strain. Bacteriophage with lytic activity against V. harveyi were isolated from prawn farm samples. Purified phage of the family Siphoviridae had a clear lytic ability and no apparent transducing properties, indicating they are appropriate for phage therapy. Phage resistance is potentially a major constraint to the use of phage therapy in aquaculture as bacteria are not completely eliminated. Phage therapy is emerging as a potential antibacterial agent that can be used to control pathogenic bacteria in aquaculture systems. The development of phage therapy for aquaculture requires initial isolation and determination of the bacteriophage host range, with subsequent creation of

  2. A selective and differential medium for Vibrio harveyi.

    OpenAIRE

    Harris, L; Owens, L; Smith, S

    1996-01-01

    A new medium, termed Vibrio harveyi agar, has been developed for the isolation and enumeration of V. harveyi. It is possible to differentiate V. harveyi colonies from the colonies of strains representing 15 other Vibrio species with this medium. This medium has been shown to inhibit the growth of two strains of marine Pseudomonas spp. and two strains of marine Flavobacterium spp. but to allow the growth of Photobacterium strains. Colonies displaying typical V. harveyi morphology were isolated...

  3. Simulated hatchery system to assess bacteriophage efficacy against Vibrio harveyi.

    Science.gov (United States)

    Raghu Patil, J; Desai, Srividya Narayanamurthy; Roy, Panchali; Durgaiah, Murali; Saravanan, R Sanjeev; Vipra, Aradhana

    2014-12-02

    Vibriosis caused by luminous Vibrio harveyi commonly contributes to poor survival in shrimp hatcheries and aquaculture ponds. Lytic bacteriophages pathogenic for V. harveyi are currently being investigated as an alternative to antibiotics to prevent vibriosis. Here, 8 bacteriophages were isolated from oysters and clams using V. harveyi strains as baiting hosts. Among these bacteriophages, 1 strain (VHP6b) identified as broadly pathogenic for 27 V. harveyi strains examined was further characterized by electron microscopy and genome sequence analysis. Phage VHP6b possessed a tail and morphology consistent with it being a member of the family Siphoviridae, and its genome and proteome were most closely related to the Vibrio phages SSP02 and MAR10. An integrase gene essential for lysogeny was not evident. The ability of bacteriophage VHP6b to protect shrimp postlarvae against vibriosis caused by V. harveyi strain VH6 was demonstrated in a model system designed to simulate typical hatchery conditions. Bacteriophage treatment improved survival of postlarvae by 40 to 60% under these conditions, so therapies based on this or other bacteriophages may be useful in shrimp hatcheries.

  4. A selective and differential medium for Vibrio harveyi.

    Science.gov (United States)

    Harris, L; Owens, L; Smith, S

    1996-01-01

    A new medium, termed Vibrio harveyi agar, has been developed for the isolation and enumeration of V. harveyi. It is possible to differentiate V. harveyi colonies from the colonies of strains representing 15 other Vibrio species with this medium. This medium has been shown to inhibit the growth of two strains of marine Pseudomonas spp. and two strains of marine Flavobacterium spp. but to allow the growth of Photobacterium strains. Colonies displaying typical V. harveyi morphology were isolated from the larval rearing water of a commercial prawn hatchery with V. harveyi agar as a primary isolation medium and were positively identified, by conventional tests, as V. harveyi. This agar displays great potential as a primary isolation medium and offers significant advantages over thiosulfate-citrate-bile salts-sucrose agar as a medium for differentiating V. harveyi from other marine and estuarine Vibrio species. PMID:8795252

  5. Pathogenic bacteriumVibrio harveyi: an endosymbiont in the marine parasitic ciliate protozoan Cryptocaryon irritans

    Institute of Scientific and Technical Information of China (English)

    QIAO Ying; WANG Jun; MAO Yong; LIU Min; CHEN Ruanni; SU Yongquan; KE Qiaozhen; HAN Kunhuang; ZHENG Weiqiang

    2017-01-01

    Vibrio harveyi, known as a pathogenic bacterium caused severe secondary bacterial infections of the large yellow croaker Larimichthys crocea, was identified as an endosymbiont in the marine parasitic ciliate protozoan Cryptocaryon irritans. Meta 16S sequencing method was used to identify the bacterial flora in C. irritans, and V. harveyi was isolated via culture-dependent method.Vibrio harveyi was observed in cytoplasm of C. irritans at the stage of tomont both by transmission electron microscopy and by Fluorescencein situ hybridization; no signal, however, was detected in nucleus area. The relationship betweenV. harveyi and C. irritans and the role of endosymbioticV. harveyi inC. irritans merit further investigation.

  6. 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 ...

  7. Autoinducers act as biological timers in Vibrio harveyi

    OpenAIRE

    Anetzberger, C.; Reiger, M.; Fekete, A.; Schell, U.; Stambrau, N.; Plener, L.; Kopka, J.; Schmitt-Kopplin, P.; Hilbi, H.; Jung, K.

    2012-01-01

    Quorum sensing regulates cell density-dependent phenotypes and involves the synthesis, excretion and detection of so-called autoinducers. Vibrio harveyi strain ATCC BAA-1116 (recently reclassified as Vibrio campbellii), one of the best-characterized model organisms for the study of quorum sensing, produces and responds to three autoinducers. HAI-1, AI-2 and CAI-1 are recognized by different receptors, but all information is channeled into the same signaling cascade, which controls a specific ...

  8. 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

  9. Heterogeneity in quorum sensing-regulated bioluminescence of Vibrio harveyi.

    Science.gov (United States)

    Anetzberger, Claudia; Pirch, Torsten; Jung, Kirsten

    2009-07-01

    Quorum sensing (QS) refers to the ability of bacterial populations to read out the local environment for cell density and to collectively activate gene expression. Vibrio harveyi, one of the best characterized model organisms in QS, was used to address the question how single cells behave within a QS-activated community in a homogeneous environment. Analysis of the QS-regulated bioluminescence of a wild type strain revealed that even at high cell densities only 69% of the cells of the population produced bioluminescence, 25% remained dark and 6% were dead. Moreover, light intensities greatly varied from cell to cell at high population density. Addition of autoinducer to a bright liquid culture of V. harveyi increased the percentage of luminescent cells up to 98%, suggesting that V. harveyi produces and/or keeps the autoinducers at non-saturating concentrations. In contrast, all living cells of a constitutive QS-active mutant (DeltaluxO) produced light. We also found that QS affects biofilm formation in V. harveyi. Our data provide first evidence that a heterogeneous population produces more biofilm than a homogeneous one. It is suggested that even a QS-committed population of V. harveyi takes advantage of heterogeneity, which extends the current view of QS-regulated uniformity.

  10. Luminescence, virulence and quorum sensing signal production by pathogenic Vibrio campbellii and Vibrio harveyi isolates.

    Science.gov (United States)

    Defoirdt, T; Verstraete, W; Bossier, P

    2008-05-01

    To study the relationship between luminescence, autoinducer production and virulence of pathogenic vibrios. Luminescence, quorum sensing signal production and virulence towards brine shrimp nauplii of 13 Vibrio campbellii and Vibrio harveyi strains were studied. Although only two of the tested strains were brightly luminescent, all of them were shown to produce the three different types of quorum sensing signals known to be produced by Vibrio harveyi. Cell-free culture fluids of all strains significantly induced bioluminescence in the cholerae autoinducer 1, autoinducer 2 and harveyi autoinducer 1 reporter strains JAF375, JMH597 and JMH612, respectively. There was no relation between luminescence and signal production and virulence towards brine shrimp. There is a large difference between different strains of Vibrio campbellii and Vibrio harveyi with respect to bioluminescence. However, this is not reflected in signal production and virulence towards gnotobiotic brine shrimp. Moreover, there seems to be no relation between quorum sensing signal production and virulence towards brine shrimp. The results presented here indicate that strains that are most brightly luminescent are not necessarily the most virulent ones and that the lower virulence of some of the strains is not due to a lack of autoinducer production.

  11. Antigenicity analysis of Vibrio harveyi TS-628 strain

    Institute of Scientific and Technical Information of China (English)

    QIN Yingxue; WANG Jun; WANG Shifeng; YAN Qingpi

    2007-01-01

    Vibrio harveyi,the major causative agent of vibriosis,affects a diverse range of marine cultured organisms over a wide geographical area.However,reports about screening the effective antigen and research on vaccines of V.harveyi are scarce.Flagellin,lipopolysaccharide (LPS) and outer membrane proteins (OMP) are major immunogenic antigens in many Gram-negative bacteria.In this study,the flagellin,OMP and LPS of the V.harveyi TS-628 strain isolated from infected groupers were extracted and Western blot analysis was used to detect the antigenicity of these extractions.Results of the Western blot assay reveal that there are four positive flagellin bands:35 kDa,38 kDa,43 kDa,and 52 kDa,of which the 43 kDa and 52 kDa bands displayed the strongest positive reaction.There are five positive OMP bands about 35 kDa,38 kDa,43 kDa,47 kDa,and 52 kDa,of which the 43 kDa appeared to have the strongest positive reaction although the other four proteins also displayed strong reactions.However,LPS is Western blot-negative.These results indicate that the 43 kDa and 52 kDa flagellin and OMP of size 43 kDa,52 kDa can be candidates for developing vaccines against V.harveyi.

  12. Effect of organic acids on shrimp pathogen, Vibrio harveyi.

    Science.gov (United States)

    Mine, Saori; Boopathy, Raj

    2011-07-01

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

  13. Quorum sensing negatively regulates chitinase in Vibrio harveyi.

    Science.gov (United States)

    Defoirdt, Tom; Darshanee Ruwandeepika, H A; Karunasagar, Indrani; Boon, Nico; Bossier, Peter

    2010-02-01

    Quorum sensing, bacterial cell-to-cell communication, regulates the virulence of Vibrio harveyi towards different hosts. Chitinase can be considered as a virulence factor because it helps pathogenic bacteria to attach to the host and to penetrate its tissues (e.g. in case of shrimp). Here, we show that quorum sensing negatively regulates chitinase in V. harveyi. Chitinolytic activity towards natural chitin from crab shells, the synthetic chitin derivative chitin azure, and fluorogenic chitin oligomers was significantly higher in a mutant in which the quorum-sensing system is completely inactivated when compared with a mutant in which the system is maximally active. Furthermore, the addition of signal molecule containing cell-free culture fluids decreased chitinase activity in a Harveyi Autoinducer 1 and Autoinducer 2-deficient double mutant. Finally, chitinase A mRNA levels were fivefold lower in the mutant in which the quorum-sensing system is maximally active when compared with the mutant in which the system is completely inactivated. [Correction added on 25 September 2009, after first online publication: the preceding sentence was corrected from 'Finally, chitinase A mRNA levels were fivefold lower in the mutant in which the quorum-sensing system is completely inactivated when compared with the mutant in which the system is maximally active.'] We argue that this regulation might help the vibrios to switch between host-associated and free-living life styles. © 2009 Society for Applied Microbiology and Blackwell Publishing Ltd.

  14. Unique and conserved genome regions in Vibrio harveyi and related species in comparison with the shrimp pathogen Vibrio harveyi CAIM 1792

    DEFF Research Database (Denmark)

    Valles, Iliana Espinoza; Vora, Gary J; Lin, Baochuan

    2015-01-01

    Vibrio harveyi CAIM 1792 is a marine bacterial strain that causes mortality in farmed shrimp in north-west Mexico, and the identification of virulence genes in this strain is important for understanding its pathogenicity. The aim of this work was to compare the V. harveyi CAIM 1792 genome....... The proteome of CAIM 1792 had higher similarity to those of other V. harveyi strains (78 %) than to those of the other closely related species Vibrio owensii (67 %), Vibrio rotiferianus (63 %) and Vibrio campbellii (59 %). Pan-genome ORFans trees showed the best fit with the accepted phylogeny based on DNA......-DNA hybridization and multi-locus sequence analysis of 11 concatenated housekeeping genes. SNP analysis clustered 34/38 genomes within their accepted species. The pangenomic and SNP trees showed that V. harveyi is the most conserved of the four species studied and V. campbellii may be divided into at least three...

  15. Aktivitas Antibakteri Ekstrak Buah Adas (Foeniculum vulgare, Mill) pada Vibrio harveyi dan Vibrio alginolyticus

    OpenAIRE

    Budianto, Budianto; Prajitno, Arief; Yuniarti, Ating

    2017-01-01

    Evaluation of natural products as a safe and effective antimicrobial agent is a scientific strategy to treat the drugresistant pathogens.Fennel(FoeniculumvulgareMill) is an herbal plant that has an active in gredient which is one of its benefit sasan antibacterial material. In thisstudy,water extract of fennel fruit determined the antibacterial activity against Vibrio harveyi and Vibrio alginolyticus using the minimum Inhibitory  Concentration Test (MIC) and paper disk diffusion method....

  16. 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.

  17. Study on the relationship of protease production and luminescence in Vibrio harveyi.

    Science.gov (United States)

    Nakayama, T; Nomura, N; Matsumura, M

    2006-07-01

    To demonstrate that Vibrio harveyi produces various types of toxins and how the production of those toxins is related with luminescence. Luminescence and toxicity of eight V. harveyi were evaluated. We demonstrated that all V. harveyi emitting luminescence were isolated from marine organisms and also showed that they were highly pathogenic when compared with culture collection V. harveyi based on cytotoxic assay test. On the contrary, V. harveyi isolated from shrimp farm showed no luminescence but showed high pathogenicity based on toxicity test. The effect of protease inhibitors on pathogenicity and luminescence was also investigated. We demonstrated that light emission of pathogenic V. harveyi remarkably decreased after addition of protease inhibitor. Furthermore, extracellular proteins from cell-free culture supernatant of luminescent and nonluminescent V. harveyi were compared using SDS-PAGE analysis. Results showed that there were differences in molecular weight and amount of proteins. Vibrio harveyi parasiting marine organisms have both luminescence and pathogenicity. Based on this study, luminescence and protease toxin activity in V. harveyi are related. Moreover, this paper clarified that V. harveyi produces various types of toxins. The current study demonstrated that V. harveyi produces two kinds of toxins, haemolysin and protease toxin. It may be clear roots of V. harveyi toxin.

  18. Biosynthesis of myristic acid in luminescent bacteria. [Vibrio harveyi

    Energy Technology Data Exchange (ETDEWEB)

    Byers, D.M.

    1987-05-01

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

  19. Quorum sensing regulates the osmotic stress response in Vibrio harveyi.

    Science.gov (United States)

    van Kessel, Julia C; Rutherford, Steven T; Cong, Jian-Ping; Quinodoz, Sofia; Healy, James; Bassler, Bonnie L

    2015-01-01

    Bacteria use a chemical communication process called quorum sensing to monitor cell density and to alter behavior in response to fluctuations in population numbers. Previous studies with Vibrio harveyi have shown that LuxR, the master quorum-sensing regulator, activates and represses >600 genes. These include six genes that encode homologs of the Escherichia coli Bet and ProU systems for synthesis and transport, respectively, of glycine betaine, an osmoprotectant used during osmotic stress. Here we show that LuxR activates expression of the glycine betaine operon betIBA-proXWV, which enhances growth recovery under osmotic stress conditions. BetI, an autorepressor of the V. harveyi betIBA-proXWV operon, activates the expression of genes encoding regulatory small RNAs that control quorum-sensing transitions. Connecting quorum-sensing and glycine betaine pathways presumably enables V. harveyi to tune its execution of collective behaviors to its tolerance to stress. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  20. Vibrio harveyi effect under survival of Litopenaeus vannamei larvae

    Directory of Open Access Journals (Sweden)

    Gabriel Aguirre-Guzmán

    2013-06-01

    Full Text Available The culture of aquatic organisms show a high relevance in the human feeding and the culture activities can create artificial conditions that increase the growth and selection of specific bacteria. Vibrio species are normal bacteria’s from microflora of penaeid shrimp, those are opportunistic pathogens that can take advantage of the ecological changes generated for the culture of aquatic organisms and which may cause diseases, low survival and economic losses in the shrimp production. The aim of this research was to determine the variation in the survival of different larval substages (nauplius, zoea I-III, mysis I-III and postlarvae 1, of Pacific white shrimp Litopenaeus vannamei exposed at three doses [103 , 105 , and 107 colony-forming unit (CFU ml-1 [ of V. harveyi, by immersion (30 min as infection method. This species generated a significant low survival in larvae (p < 0.05 only in high doses (105 and 107 CFU ml-1 , where higher doses show the lowest values of survival. Larval substages and postlarvae 1 of shrimp showed sensitivity associate to the increase of Vibrio doses and this sensitivity decreased with the growth of larval substages and postlarvae 1. This information has high significance for the fisheries and aquaculture industry, which help to generate strategies to reduce the effects of V. harveyi with positive effect in growth and survival of the shrimp larvae and postlarvae 1.

  1. Autoinducers act as biological timers in Vibrio harveyi.

    Science.gov (United States)

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

    2012-01-01

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

  2. Autoinducers act as biological timers in Vibrio harveyi.

    Directory of Open Access Journals (Sweden)

    Claudia Anetzberger

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

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

    Science.gov (United States)

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

    2002-01-01

    Physiological 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, direct viable counts, actively respiring cell counts, nucleoid-containing cell counts, and total counts were determined. Vibrio harveyi incubated at 22 degrees C in nutrient-limited artificial seawater (ASW) became nonculturable after approximately 62 and 45 d in microcosms of 35 ppt and 10 ppt ASW, respectively. In contrast, V. fischeri became nonculturable at approximately 55 and 31 d in similar microcosms. Recovery of both culturability and luminescence of cells in the viable but nonculturable state was achieved by addition of nutrient broth or nutrient broth supplemented with a carbon source, including luminescence-stimulating compounds. Temperature upshift from 22 degrees C to 30 degrees C or 37 degrees C did not result in recovery from nonculturability. The study confirms entry of V. harveyi and V. fischeri into the viable but nonculturable state under low-nutrient conditions and demonstrates nutrient-dependent resuscitation from this state. This study confirms loss of luminescence of V. harveyi and V. fischeri on entry into the viable but nonculturable state and suggests that enumeration of luminescent cells in water samples may be a rapid method to deduce the nutrient status of a water sample.

  4. Vibrio harveyi modulated gene expression in Penaeus monodon and Fenneropenaeus indicus

    Digital Repository Service at National Institute of Oceanography (India)

    Nayak, S.

    : Gammaproteobacteria Order : Vibrionales Family : Vibrionaceae Genus : Vibrio Species : harveyi Vibrio harveyi is a Gram-negative bioluminescent marine bacterium. It is both a free- living, as well as a symbiont with many... marine animals. In free-living form, it can be found throughout the water column, in the sediment as well as on the exterior surfaces of marine organisms. It causes systemic infection resulting in mortalities in larvae and post-larvae, which sometimes...

  5. Potensi Ekstrak Daun Binahong (Anredera cordifolia) Sebagai Penghambat Bakteri Vibrio harveyi

    OpenAIRE

    Gde Raka Angga Kartika; Sri Andayani; Soelistyowati Soelistyowati

    2016-01-01

    Binahong (Anredera cordifolia) is a plant that can treat various kinds of diseases, because this plant has a high antioxidant content and as an antibacterial and antiviral. Vibriosis disease caused by the bacterium Vibrio harveyi is a serious problem in marine and brackish culture, this disease can cause death for shrimp and fish that farmed in marine or brackish. This study aims to determine the potential of using leaf extract Binahong with different concentrations as Vibrio harveyi inhibiti...

  6. Bacteroides species produce Vibrio harveyi autoinducer 2-related molecules.

    Science.gov (United States)

    Antunes, Luis Caetano Martha; Ferreira, Lívia Queiroz; Ferreira, Eliane Oliveira; Miranda, Karla Rodrigues; Avelar, Kátia Eliane Santos; Domingues, Regina Maria Cavalcanti Pilotto; Ferreira, Maria Candida de Souza

    2005-10-01

    Quorum sensing is a density-dependent gene regulation mechanism that has been described in many bacterial species in the last decades. Bacteria that use quorum sensing as part of their gene regulation circuits produce molecules called autoinducers that accumulate in the environment and activate target genes in a quorum-dependent way. Some specific clues led us to hypothesize that Bacteroides species can produce autoinducers and possess a quorum sensing system. First, Bacteroides are anaerobic bacteria that are frequently involved in polymicrobial infections. These infections often involve Pseudomonas aeruginosa and Staphylococcus aureus, two of the best understood examples of bacteria that employ quorum sensing systems as part of their pathogenesis. Also, studies have detected the presence of a quorum sensing gene involved in the production of autoinducers in Porphyromonas gingivalis, a species closely related to the Bacteroides genus. These and other evidences prompted us to investigate if Bacteroides strains could produce autoinducer molecules that could be detected by a Vibrio harveyi reporter system. In this paper, we show that supernatants of B. fragilis, B. vulgatus and B. distasonis strains are able to stimulate the V. harveyi quorum sensing system 2. Also, we were able to demonstrate that the stimulation detected is due to the production of autoinducer molecules and not the growth of reporter strains after addition of supernatant. Moreover, the phenomenon observed does not seem to represent the degradation of repressors possibly present in the culture medium used. We could also amplify bands from some of the strains tested using primers designed to the luxS gene of Escherichia coli. Altogether, our results show that B. fragilis, B. vulgatus and B. distasonis (but possibly some other species) can produce V. harveyi autoinducer 2-related molecules. However, the role of such molecules in the biology of these organisms remains unknown.

  7. Improved quorum sensing capacity by culturing Vibrio harveyi in microcapsules.

    Science.gov (United States)

    Gao, Meng; Song, Huiyi; Liu, Xiudong; Yu, Weiting; Ma, Xiaojun

    2016-04-01

    Microcapsule entrapped low density cells with culture (ELDCwc), different from free cell culture, conferred stronger stress resistance and improved cell viability of microorganisms. In this paper, the quorum sensing (QS) system of Vibrio harveyi was used to investigate changes when cells were cultured in microcapsules. Cells in ELDCwc group grew into cell aggregates, which facilitated cell-cell communication and led to increased bioluminescence intensity. Moreover, the luxS-AI-2 system, a well-studied QS signal pathway, was detected as both luxS gene and the AI-2 signaling molecule, and the results were analyzed with respect to QS capacity of unit cell. The V. harveyi of ELDCwc also showed higher relative gene expression and stronger quorum sensing capacity when compared with free cells. In conclusion, the confined microcapsule space can promote the cell aggregates formation, reduce cell-cell communication distance and increase local concentration of signal molecule, which are beneficial to bacterial QS. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  8. Competitive Dominance by a Bacteriocin-Producing Vibrio harveyi Strain.

    Science.gov (United States)

    Hoyt, P R; Sizemore, R K

    1982-09-01

    Vibrio (Beneckea) harveyi, a bioluminescent marine bacterium, has been shown to produce a bacteriocin-like substance the production of which is mediated by a plasmid. This substance is assumed to be proteinaceous because of its sensitivity to certain proteolytic enzymes. It is stable at low temperatures and can be concentrated by ammonium sulfate precipitation or negative-pressure dialysis. The molecular weight of the bacteriocin was determined to be 2.4 x 10 by molecular exclusion chromatography. Competition experiments indicated that bacteriocin-producing strains predominated over cured variants of the same strain in broth culture experiments. We studied several environmental parameters (pH, salinity, temperature, nutrient concentration) to determine their effects on the competitive advantage bestowed on a bacteriocin-producing strain. Under simulated free-living conditions, no competitive advantage attributable to bacteriocin production was observed. In a simulated enteric habitat, a bacteriocin-producing strain showed dramatic (>90%) inhibition of the sensitive strain within 24 h.

  9. Development and efficacy of an attenuated Vibrio harveyi vaccine candidate with cross protectivity against Vibrio alginolyticus.

    Science.gov (United States)

    Hu, Yong-hua; Deng, Tian; Sun, Bo-guang; Sun, Li

    2012-06-01

    Vibrio harveyi is a Gram-negative bacterial pathogen that can infect a wide range of marine animals. In previous studies, we have reported a virulent V. harveyi strain, T4D. In the present study, an attenuated mutant of T4D, T4DM, was obtained by selection of rifampicin resistance. Compared to the wild type, T4DM was different in whole-cell protein profile and much slower in growth rate when cultured in stress conditions caused by iron depletion. Virulence analysis showed that compared to T4D, T4DM exhibited a dramatically increased median lethal dose, impaired tissue dissemination capacity, defective hemolytic activity, and significantly reduced resistance against the killing effect of host serum. To examine the potential of T4DM as a live attenuated vaccine, Japanese flounder (Paralichthys olivaceus) were vaccinated with T4DM via intraperitoneal injection or immersion. The results showed that at one and two months post-vaccination, fish administered with T4DM via both approaches, in particular that of immersion, were effectively protected against not only V. harveyi but also Vibrio alginolyticus, another important fish pathogen. Microbiological analysis showed that following immersion vaccination, T4DM was recovered from the internal organs of the vaccinated fish in a time-dependent manner within the first 6 days post-vaccination. Serum antibodies against V. harveyi and V. alginolyticus were detected in T4DM-vaccinated fish, and, compared to serum from control fish, serum from T4DM-vaccinated fish was significantly enhanced in bactericidal activity. These results indicate that T4DM is an attenuated strain with residual infectivity and that T4DM can induce effective cross-species protection against both V. harveyi and V. alginolyticus when used as a live immersion vaccine. Copyright © 2012 Elsevier Ltd. All rights reserved.

  10. Vibrio lentus protects gnotobiotic sea bass (Dicentrarchus labrax L.) larvae against challenge with Vibrio harveyi.

    Science.gov (United States)

    Schaeck, M; Duchateau, L; Van den Broeck, W; Van Trappen, S; De Vos, P; Coulombet, C; Boon, N; Haesebrouck, F; Decostere, A

    2016-03-15

    Due to the mounting awareness of the risks associated with the use of antibiotics in aquaculture, treatment with probiotics has recently emerged as the preferred environmental-friendly prophylactic approach in marine larviculture. However, the presence of unknown and variable microbiota in fish larvae makes it impossible to disentangle the efficacy of treatment with probiotics. In this respect, the recent development of a germ-free culture model for European sea bass (Dicentrarchus labrax L.) larvae opened the door for more controlled studies on the use of probiotics. In the present study, 206 bacterial isolates, retrieved from sea bass larvae and adults, were screened in vitro for haemolytic activity, bile tolerance and antagonistic activity against six sea bass pathogens. Subsequently, the harmlessness and the protective effect of the putative probiotic candidates against the sea bass pathogen Vibrio harveyi were evaluated in vivo adopting the previously developed germ-free sea bass larval model. An equivalence trial clearly showed that no harmful effect on larval survival was elicited by all three selected probiotic candidates: Bacillus sp. LT3, Vibrio lentus and Vibrio proteolyticus. Survival of Vibrio harveyi challenged larvae treated with V. lentus was superior in comparison with the untreated challenged group, whereas this was not the case for the larvae supplemented with Bacillus sp. LT3 and V. proteolyticus. In this respect, our results unmistakably revealed the protective effect of V. lentus against vibriosis caused by V. harveyi in gnotobiotic sea bass larvae, rendering this study the first in its kind. Copyright © 2016. Published by Elsevier B.V.

  11. The effect of copper concentration on the virulence of pathogenic Vibrio harveyi.

    Science.gov (United States)

    Nakayama, T; Nomura, N; Matsumura, M

    2007-05-01

    To demonstrate the influence of copper on luminescence and toxin production in Vibrio harveyi. The effect of copper concentration on the expression of both luminescence and toxin of V. harveyi was investigated. Copper concentration of less than 40 ppm had no effect on the growth. While V. harveyi cultured with 40 ppm copper concentration showed decreased luminescence as measured by spectrofluorophotometer and as observed. LuxD gene, which is related to luminescence expression, was monitored using real-time RT-PCR. Result showed that the concentration of cDNA coding for luxD was lower in V. harveyi with copper. Toxic activity against both HeLa cells and shrimp haemocytes was also lower in the culture supernatant of V. harveyi grown with 40 ppm copper concentration. Moreover, V. harveyi extracellular proteins were analysed using SDS-PAGE. Results showed that culture supernatant from V. harveyi grown without copper had thicker band indicating a higher concentration of the putative cysteine protease, one of the major toxin of V. harveyi. This study proved that both luminescence and toxin were repressed by copper. The current study demonstrated that copper inhibited expression of phenotype of V. harveyi. Furthermore, it may inhibit quorum sensing of V. harveyi.

  12. Luciferase inactivation in the luminous marine bacterium Vibrio harveyi.

    Science.gov (United States)

    Reeve, C A; Baldwin, T O

    1981-06-01

    Luciferase was rapidly inactivated in stationary-phase cultures of the wild type of the luminous marine bacterium Vibrio harveyi, but was stable in stationary-phase cultures of mutants of V. harveyi that are nonluminous without exogenous aldehyde, termed the aldehyde-deficient mutants. The inactivation in the wild type was halted by cell lysis and was slowed or stopped by O2 deprivation or by addition of KCN and NaF or of chloramphenicol. If KCN and NaF or chloramphenicol were added to a culture before the onset of luciferase inactivation, then luciferase inactivation did not occur. However, if these inhibitors were added after the onset of luciferase inactivation, then luciferase inactivation continued for about 2 to 3 h before the inactivation process stopped. The onset of luciferase inactivation in early stationary-phase cultures of wild-type cell coincided with a slight drop in the intracellular adenosine 5'-triphosphate (ATP) level from a relatively constant log-phase value of 20 pmol of ATP per microgram of soluble cell protein. Addition of KCN and NaF to a culture shortly after this drop in ATP caused a rapid decrease in the ATP level to about 4 pmol of ATP per microgram whereas chloramphenicol added at this same time caused a transient increase in ATP level to about 25 pmol/microgram. The aldehyde-deficient mutant (M17) showed a relatively constant log-phase ATP level identical with that of the wild-type cells, but rather than decreasing in early stationary phase, the ATP level increased to a value twice that in log-phase cells. We suggest that the inactivation of luciferase is dependent on the synthesis of some factor which is produced during stationary phase and is itself unstable, and whose synthesis is blocked by chloramphenicol or cyanide plus fluoride.

  13. Luminescence, virulence and quorum sensing signal production by pathogenic Vibrio campbellii and Vibrio harveyi isolates

    OpenAIRE

    Defoirdt, T.; Verstraete, W.; Bossier, P.

    2008-01-01

    Aims: To study the relationship between luminescence, autoinducer production and virulence of pathogenic vibrios.Methods and Results: Luminescence, quorum sensing signal production and virulence towards brine shrimp nauplii of 13 Vibrio campbellii and Vibrio harveyi strains were studied. Although only two of the tested strains were brightly luminescent, all of them were shown to produce the three different types of quorum sensing signals known to be produced by Vibrio harveyi. Cell-free cultu...

  14. Inhibitory effects of Bacillus probionts on growth and toxin production of Vibrio harveyi pathogens of shrimp.

    Science.gov (United States)

    Nakayama, T; Lu, H; Nomura, N

    2009-12-01

    To investigate the effects of Bacillus subtilis, Bacillus licheniformis and Bacillus megaterium in terms of toxin and growth of pathogenic Vibrio harveyi. Three Bacillus probionts were isolated from probiotic BZT aquaculture and identified using a 16S rDNA sequence. Growth inhibition assay showed that supernatants from the 24-h culture of three Bacillus species were able to inhibit the growth of V. harveyi (LMG 4044); B. subtilis was the most effective based on the well diffusion method. Results of a liquid culture model showed that B. subtilis was also widely effective in inhibiting three strains of V. harveyi (isolated from Thailand, the Philippines and LMG 4044), and that both B. licheniformis and B. megaterium inhibit the growth of V. harveyi isolated from the Philippines. Moreover, a haemolytic activity assay demonstrated that V. harveyi (IFO 15634) was significantly decreased by the addition of B. licheniformis or B. megaterium supernatant. Bacillus subtilis inhibited Vibrio growth, and both B. licheniformis and B. megaterium suppressed haemolytic activity in Vibrio. The cell-free supernatants produced by Bacillus probionts inhibit Vibrio disease, and Bacillus probionts might have an influence on Vibrio cell-to-cell communications.

  15. Potensi Ekstrak Daun Binahong (Anredera cordifolia Sebagai Penghambat Bakteri Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    Gde Raka Angga Kartika

    2016-07-01

    Full Text Available Binahong (Anredera cordifolia is a plant that can treat various kinds of diseases, because this plant has a high antioxidant content and as an antibacterial and antiviral. Vibriosis disease caused by the bacterium Vibrio harveyi is a serious problem in marine and brackish culture, this disease can cause death for shrimp and fish that farmed in marine or brackish. This study aims to determine the potential of using leaf extract Binahong with different concentrations as Vibrio harveyi inhibiting bacterial growth in vitro. The method used is to test the Minimum inhibitory concentration (MIC to determine minimum levels inhibit the growth of Vibrio harveyi and Minimum Bactericidal Concentration (MBC using paper disc. The results showed the use of leaf extract Binahong (Anredera cordifolia with different concentrations significant effect on the growth of Vibrio harveyi in vitro. Binahong leaf extract (Anredera cordifolia with a concentration of 3%, 5%, 7%, 9%, 11% and 13% is only bacteriostatic which inhibits the growth of bacteria Vibrio harveyi with the best concentration obtained is equal to 13%.

  16. The application of bioflocs technology to protect brine shrimp (Artemia franciscana) from pathogenic Vibrio harveyi.

    Science.gov (United States)

    Crab, R; Lambert, A; Defoirdt, T; Bossier, P; Verstraete, W

    2010-11-01

    To study the potential biocontrol activity of bioflocs technology. Glycerol-grown bioflocs were investigated for their antimicrobial and antipathogenic properties against the opportunistic pathogen Vibrio harveyi. The bioflocs did not produce growth-inhibitory substances. However, bioflocs and biofloc supernatants decreased quorum sensing-regulated bioluminescence of V. harveyi. This suggested that the bioflocs had biocontrol activity against this pathogen because quorum sensing regulates virulence of vibrios towards different hosts. Interestingly, the addition of live bioflocs significantly increased the survival of gnotobiotic brine shrimp (Artemia franciscana) larvae challenged to V. harveyi. Bioflocs grown on glycerol as carbon source inhibit quorum sensing-regulated bioluminescence in V. harveyi and protect brine shrimp larvae from vibriosis. The results presented in this study indicate that in addition to water quality control and in situ feed production, bioflocs technology could help in controlling bacterial infections within the aquaculture pond. © 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for Applied Microbiology.

  17. GENETIC DIVERSITY OF Vibrio harveyi ISOLATED FROM TIGER PRAWN Penaeus monodon HATCHERIES AND GROW OUT PONDS

    OpenAIRE

    Muliani Muliani; Ince Ayu Khairana Kadriah; Andi Parenrengi; Sulaeman Sulaeman

    2006-01-01

    ibrio harveyi is known as one among the most harmful bacteria infecting tiger prawn at every stage of its life’s. The present research was aimed to reveal the genetic diversity of Vibrio harveyi isolated from tiger prawn (Penaeus monodon) culture. The samples of bacteria were collected from hatchery (brood-stock, larvae, natural feed, artemia, and larval rearing water) and grow-out (juveniles, water, shrimp, sediment, plankton, crab, mollusc, microalgae, and wild fish).  The taxonomic identif...

  18. Effect of combined function of temperature and water activity on the growth of Vibrio harveyi

    OpenAIRE

    Zhou,Kang; Gui,Meng; Li,Pinglan; Xing,Shaohua; Cui,Tingting; Peng,Zhaohui

    2012-01-01

    Vibrio harveyi is considered as a causative agent of the systemic disease, vibriosis, which occurs in many biological fields. The effects of temperatures (12.9-27.1 ºC) and water activity (NaCl% 0.6%-3.4%) on V. harveyi were investigated. The behavior and growth characteristics of V. harveyi was studied and modeled. Growth curves were fitted by using Gompertz and Baranyi models, and the Baranyi model showed a better fittness. Then, the maximum growth rates (µmax) and lag phase durations (LPD,...

  19. GENETIC DIVERSITY OF Vibrio harveyi ISOLATED FROM TIGER PRAWN Penaeus monodon HATCHERIES AND GROW OUT PONDS

    Directory of Open Access Journals (Sweden)

    Muliani Muliani

    2006-06-01

    Full Text Available ibrio harveyi is known as one among the most harmful bacteria infecting tiger prawn at every stage of its life’s. The present research was aimed to reveal the genetic diversity of Vibrio harveyi isolated from tiger prawn (Penaeus monodon culture. The samples of bacteria were collected from hatchery (brood-stock, larvae, natural feed, artemia, and larval rearing water and grow-out (juveniles, water, shrimp, sediment, plankton, crab, mollusc, microalgae, and wild fish.  The taxonomic identification of Vibrio spp.was performed based on the physiological and biochemical characteristic following the isolation by Thiosulphate Citrate Bile-salt Sucrose Agar (TCBSA media. Amplified Ribosomoal DNA Restriction Analysis (ARDRA for 16S-rRNA analysis and Macrorestriction Fragment Length Polymorphism (MLFP analysis using Pulsed Field Gel Electrophoresis (PFGE were applied to reveal the genetic diversity of V. harveyi. According to the taxonomic identification, of 361 isolates of Vibrio spp., 129 isolates (35.7% were identified as V. harveyi. The result of ARDRA analysis showed that the 16S-rRNA gene of V. harveyi digested by RsaI and HhaI enzyme, each generated three and four identical fragments respectively for the all samples. These meaned that ARDRA could not reveal any genetic variation on V. harveyi. The size range of all DNA fragment was less than 500 bp. This result indicated that the high genetic diversity of V. harveyi was revealed by MFLP-PFGE analysis. DNA fragment of V. harveyi was digested by NotI enzyme.

  20. Unexpected photoreactivation of Vibrio harveyi bacteria living in ionization environment

    International Nuclear Information System (INIS)

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

    2011-01-01

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

  1. Detection of Vibrio harveyi using hemolysin primer in tiger shrimp Penaeus monodon

    Directory of Open Access Journals (Sweden)

    Irma Suriyani

    2015-05-01

    Full Text Available ABSTRACT This study was aimed to analyze the sensitivity and ability of primer hemolysin in detecting pathogenetic Vibrio on tiger shrimp post-larvae (PL exposed under different exposure times in media inoculated with Vibrio harveyi. The PL of tiger shrimp were infected with 106 cfu/mL of V. harveyi by immersion method for three, six, 12, 24, 48 and 72 hours. The presence of hemolisin genes was detected by PCR techniques. The electrophoresis detected narrow hemolysin genes after PL were exposed for three and six hours. Clear visible bands of DNA Vibrio were observed for 12 hours of exposure. In contrast, no detected hemolysin gene of Vibrio was observed for PL exposed within 24, 48, and 72 hours. The rapid detection on Vibrio pathogenic for tiger shrimp PL should be conducted within three to 12 hours of exposure. No recommendation in utilizing this rapid detection for tiger shrimp PL exposed beyond 12 hours of V. harveyi. Keywords: specific primer, luminous Vibrio bacteria, pathogenic, PCR method, hemolysin gene  ABSTRAK Penelitian ini bertujuan untuk mengetahui kemampuan atau sensitivitas primer hemolisin dalam mendeteksi Vibrio patogen dengan lama pemaparan berbeda. Penelitian ini dilakukan dengan menginfeksikan Vibrio harveyi pada benur udang dengan metode perendaman pada konsentrasi 106 cfu/mL. Pengambilan sampel dilakukan pada waktu tiga, enam, 12, 24, 48, dan 72 jam pascainfeksi. Keberadaan gen hemolisin pada bakteri V. harveyi dideteksi menggunakan teknik polymerase chain reaction (PCR. Hasil elektroforesis memperlihatkan bahwa pada pemaparan tiga dan enam jam keberadaan gen hemolisin dari bakteri Vibrio patogen yang diinfeksikan sudah dapat terdeteksi pada benur walaupun masih terlihat tipis. Pada pemaparan 12 jam terlihat sangat jelas pita-pita DNA dari bakteri patogen. Sedangkan pada pemaparan 24, 48, dan 72 jam sudah tidak terdeteksi lagi gen hemolisin dari bakteri Vibrio. Hal ini diduga disebabkan terjadinya penurunan populasi

  2. Molecular variations in Vibrio alginolyticus and V. harveyi in shrimp-farming systems upon stress

    OpenAIRE

    Santhyia,Anix Vivek; Mulloorpeedikayil,Rosalind George; Kollanoor,Riji John; Jeyaseelan,Prince M.J.

    2015-01-01

    A study was performed to investigate the genomic variations in the shrimp farm isolates of Vibrio alginolyticus and V. harveyi when the isolates were subjected to environmental stress. Samples of shrimps, water and sediment were collected from Southern Indian coastal shrimp farms. Vibrio isolates were biochemically identified and confirmed using 16S rDNA and gyrB gene specific PCR. The bacterial strains were genotyped by PCR fingerprinting using GTG(5) and IS (Insertion Sequence) primers. Sev...

  3. Comparative analysis of the survival and gene expression of pathogenic strains Vibrio harveyi after starvation.

    Science.gov (United States)

    Sun, Jingjing; Gao, Xiaojian; Qun, Jiang; Du, Xuedi; Bi, Keran; Zhang, Xiaojun; Lin, Li

    2016-11-01

    This study aimed to evaluate the survival and gene expression of Vibrio harveyi under starvation conditions. The microcosms V. harveyi were incubated in sterilized seawater for 4 weeks at room temperature. Overall, the cell numeration declined rapidly about 10 3 CFU/ml during starvation, with a tiny rebound at day 21. Scanning electron microscopy revealed that rod-shaped cells became sphere with a rippled cell surface. By polymerase chain reaction (PCR) assay, nine genes, named luxR, toxR, vhhB, flaA, topA, fur, rpoS, mreB and ftsZ, were detected in the non-starved cells. In the starved cells, the expression levels of the detected genes declined substantially ranging from 0.005-fold to 0.028-fold compared to the non-starved cells performed by reverse transcription quantitative real-time PCR with 16S rRNA as the internal control. In the recovering cells, the expression levels of the detected genes, except luxR and mreB, were upregulated dramatically compared to the wild, especially topA (23.720-fold), fur (39.400-fold) and toxR (9.837-fold), validating that the expressions of both the metabolism and virulence genes were important for growth and survival of V. harveyi. The results may shed a new light on understanding of stress adaptation in bacteria. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Oral administration of formalin killed Vibrio anguillarum cells improves growth and protection against challenge with Vibrio harveyi in banana shrimp.

    Science.gov (United States)

    Patil, P K; Gopal, C; Panigrahi, A; Rajababu, D; Pillai, S M

    2014-03-01

    Larval rearing in hatcheries and highly intensive grow-out culture practices followed in shrimp production systems favour the growth of potential pathogenic bacterial loads. This study reports the efficacy of formalin-killed vibrio bacterin on growth, survival and protection to challenge with virulent Vibrio harveyi and Vibrio anguillarum in juveniles of banana shrimp Fenneropenaeus merguiensis. Postlarvae 15 (0·24 ± 0·01 g) were administered orally in different concentrations of bacterial preparation (0, 10(6) , 10(8) , 10(10) and 10(12 ) CFU kg(-1) feed) for a period of 6 weeks. Physicochemical and microbial quality of water in larval rearing tanks, and growth and survival of the postlarvae were monitored at regular intervals, and body composition was estimated at the end of the experiment. Shrimps were challenged with V. harveyi and V. anguillarum, and cumulative mortality was calculated. The group receiving 10(8)  CFU kg(-1) feed showed highest average weight gain (162·66 ± 22·94 mg) and survival (90·33 ± 4·5%) and lowest cumulative mortality following the challenge with V. anguillarum (26%) and V. harveyi (36·67%). The results of the study suggest that formalized vibrio administered orally to F. merguiensis postlarvae could induce both homologous and heterologous protection against V. anguillarum and V. harveyi. 'Vaccination' of shrimp postlarvae at hatcheries would help in preventing the losses due to vibriosis and the most susceptible stages of shrimp development. The study demonstrates the cross-protection offered by the oral feeding of formalin-killed Vibrio anguillarum against pathogenic V. harveyi challenge at the early developmental stages of banana shrimp, Fenneropenaeus merguiensis. © 2013 The Society for Applied Microbiology.

  5. Partial characterization of a novel bacteriophage of Vibrio harveyi isolated from shrimp culture ponds in Thailand.

    Science.gov (United States)

    Pasharawipas, Tirasak; Thaikua, Surasak; Sriurairatana, Siriporn; Ruangpan, Lila; Direkbusarakum, Sataporn; Manopvisetcharean, Jaroon; Flegel, Timothy W

    2005-12-01

    A bacteriophage was isolated together with Vibrio harveyi (VH) 1114 a from a black tiger shrimp-rearing pond in Thailand. By negative staining transmission electron microscopy (TEM), the phage had an icosahedral head (diameter 60-62 nm), a rigid, non-contractile tail (9-10 nm x 100-120 nm) without a collar or terminal fibers and a genome of double stranded DNA of approximately 80 kb as determined by analysis of restriction enzyme digestion fragments. Since these features would place it in the family Siphoviridae, it was tentatively named V. harveyi siphoviridae-like phage or VHS1. VHS1 could also infect two VH reference strains LMD 22.30 and LMD 80.33 (=ATCC 14126) but yielded smaller plaques than with VH1114. The phage tolerated temperatures as high as 60 degrees C for up to 2h and overnight exposure to a broad range of pH. VHS1 lysogens of VH1114 were unstable, contained unaltered VHS1 DNA, were immune to VHS1 lysis and spontaneously released VHS1 in liquid cultures. Approximately 20 kb of the genome has been sequenced and deposited at GenBank but it mostly showed no significant homology with existing sequences in the database.

  6. Vibrio harveyi as a causative agent of the White Syndrome in tropical stony corals.

    Science.gov (United States)

    Luna, Gian Marco; Bongiorni, Lucia; Gili, Claudia; Biavasco, Francesca; Danovaro, Roberto

    2010-02-01

    We investigated bacterial assemblages associated with corals displaying symptoms of the 'White Syndrome' (WS), a general term used for indicating the appearance of bands, spots or patches of tissue loss, which is devastating wide areas of tropical ecosystems worldwide. We collected WS-diseased (n = 15) and healthy (n = 15) corals from the natural reef (Indonesia, Indian Ocean) and from four large public aquaria. By using culture-dependent and culture-independent techniques, we found that a large fraction (73%) of the investigated WS events was associated with the presence of a high bacterial abundance and, specifically, of Vibrio spp. Vibrio harveyi, a pathogen of many marine organisms and recently involved in coral Yellow Band disease, was the most represented species, being recovered from five out of 15 diseased corals. In experimental infection assays, two V. harveyi strains, isolated from diseased corals, were inoculated on a total of 62 healthy colonies of Pocillopora damicornis. WS signs appeared in 57 corals, confirming the ability of V. harveyi strains to induce the disease. We conclude that V. harveyi is one of the coral pathogens involved in the appearance of WS. However, not all of the investigated WSs were associated to V. harveyi detection, nor to other Vibrio species (such as V. coralliilyticus), which supports the hypothesis that WS is not caused exclusively by Vibrio spp., but rather can have a multifactorial aetiology, or can represent a group of diseases caused by a variety of agents. Further investigations to identify specific virulence traits will contribute to the understanding of the role of V. harveyi in WS pathogenesis. © 2009 Society for Applied Microbiology and Blackwell Publishing Ltd.

  7. Regulation of Metalloprotease Gene Expression in Vibrio vulnificus by a Vibrio harveyi LuxR Homologue

    Science.gov (United States)

    Shao, Chung-Ping; Hor, Lien-I

    2001-01-01

    Expression of the Vibrio vulnificus metalloprotease gene, vvp, was turned up rapidly when bacterial growth reached the late log phase. A similar pattern of expression has been found in the metalloprotease gene of Vibrio cholerae, and this has been shown to be regulated by a Vibrio harveyi LuxR-like transcriptional activator. To find out whether a LuxR homologue exists in V. vulnificus, a gene library of this organism was screened by colony hybridization using a probe derived from a sequence that is conserved in various luxR-like genes of vibrios. A gene containing a 618-bp open reading frame was identified and found to be identical to the smcR gene of V. vulnificus reported previously. An isogenic SmcR-deficient (RD) mutant was further constructed by an in vivo allelic exchange technique. This mutant exhibited an extremely low level of vvp transcription compared with that of the parent strain. On the other hand, the cytolysin gene, vvhA, was expressed at a higher level in the RD mutant than in the parent strain during the log phase of growth. These data suggested that SmcR might not only be a positive regulator of the protease gene but might also be involved in negative regulation of the cytolysin gene. Virulence of the RD mutant in either normal or iron-overloaded mice challenged by intraperitoneal injection was comparable to that of the parent strain, indicating that SmcR is not required for V. vulnificus virulence in mice. PMID:11157950

  8. Vibrio harveyi Adheres to and Penetrates Tissues of the European Abalone Haliotis tuberculata within the First Hours of Contact

    OpenAIRE

    Cardinaud, Marion; Barbou, Annaïck; Capitaine, Carole; Bidault, Adeline; Dujon, Antoine Marie; Moraga, Dario; Paillard, Christine

    2014-01-01

    International audience; Vibrio harveyi is a marine bacterial pathogen responsible for episodic epidemics generally associated with massive mortalities in many marine organisms, including the European abalone Haliotis tuberculata. The aim of this study was to identify the portal of entry and the dynamics of infection of V. harveyi in the European abalone. The results indicate that the duration of contact be-tween V. harveyi and the European abalone influences the mortality rate and precocity. ...

  9. Regulation of expression of Na+ -translocating NADH:quinone oxidoreductase genes in Vibrio harveyi and Klebsiella pneumoniae.

    Science.gov (United States)

    Fadeeva, Maria S; Yakovtseva, Evgenia A; Belevich, Galina A; Bertsova, Yulia V; Bogachev, Alexander V

    2007-10-01

    The expression of genes encoding sodium-translocating NADH:quinone oxidoreductase (Na(+)-NQR) was studied in the marine bacterium Vibrio harveyi and in the enterobacterium Klebsiella pneumoniae. It has been shown that such parameters as NaCl concentration, pH value, and presence of an uncoupler in the growth media do not influence significantly the level of nqr expression. However, nqr expression depends on the growth substrates used by these bacteria. Na(+)-NQR is highly repressed in V. harveyi during anaerobic growth, and nqr expression is modulated by electron acceptors and values of their redox potentials. The latter effect was shown to be independent of the ArcAB regulatory system.

  10. Pathogenicity Assay of Vibrio harveyi in Tiger Shrimp Larvae Employing Rifampicin-Resistant as A Molecular Marker

    Directory of Open Access Journals (Sweden)

    . Widanarni

    2007-12-01

    Full Text Available Rifampicin-resistant marker was employed as a reporter to assay pathogenicity of Vibrio harveyi  in shrimp larvae.  V. harveyi M. G3 and G7 that difference not schizotyping as shown by Pulsed-Filed Gel Electrophoresis (PFGE used in this study. Spontaneous mutation was conducted to generate V. harveyi resistant to rifampicin. Two groups of shrimp post-larvae (PL5 were immersed for 30 min in 106 CFU/ml of mutants and wild type of V. harveyi, respectively; and then placed in a 2 liter shrimp rearing tank for five days. A control group was immersed in sterile seawater. Growth curve analysis and pathogenicity assay of V. harveyi  showed that each of the V. harveyi mutant exhibited almost identical profiles to that of the wild type parental strain and did not show alteration in their pathogenicity. Sample from dead shrimp larvae showed that the dead shrimp larvae were infected by V. harveyi RfR, indicated that rifampicin-resistant marker effective as a reporter to assay pathogenicity of Vibrio harveyi in shrimp larvae. Key words: shrimp larvae, Vibrio harveyi, rifampicin-resistant, molecular marker

  11. 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...... in Crete, Greece....

  12. In vitro interactions of Acanthamoeba castellanii Neff and Vibrio harveyi.

    Science.gov (United States)

    Reyes-Batlle, María; Martín-Rodríguez, Alberto J; López-Arencibia, Atteneri; Sifaoui, Ines; Liendo, Aitor Rizo; Bethencourt Estrella, Carlos J; García Méndez, Ana B; Chiboub, Olfa; Hajaji, Soumaya; Valladares, Basilio; Martínez-Carretero, Enrique; Piñero, José E; Lorenzo-Morales, Jacob

    2017-12-01

    Free-living amoebae (FLA) are opportunistic protozoa widely distributed in the environment. They are frequently found in water and soil samples, but they have also been reported to be associated with bacterial human pathogens such as Legionella spp. Campylobacter spp or Vibrio cholerae among others. Including within Vibrio spp. V. harveyi (Johnson and Shunk, 1936) is a bioluminescent marine bacteria which has been found swimming freely in tropical marine waters, being part of the stomach and intestine microflora of marine animals, and as both a primary and opportunistic pathogen of marine animals. Our aim was to study the interactions between Vibrio harveyi and Acanthamoeba castellanii Neff. Firstly, in order to analyze changes in it cultivability, V. harveyi was coincubated with A. castellanii Neff axenic culture and with Acanthamoeba Conditioned Medium (ACM) at different temperatures in aerobic conditions. Interestingly, at 4 °C and 18-20 °C bacteria were still cultivable in marine agar, at 28 °C, in aerobic conditions, but there weren't significant differences comparing with the controls. We also noted an enhanced migration of Acanthamoeba toward V. harveyi on non-nutrient agar plates compared to controls with no bacteria. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Phenotypic and molecular typing of Vibrio harveyi isolates and their pathogenicity to tiger shrimp larvae.

    Science.gov (United States)

    Alavandi, S V; Manoranjita, V; Vijayan, K K; Kalaimani, N; Santiago, T C

    2006-11-01

    The objective of the present study was to identify the biotype(s) and molecular type(s) of Vibrio harveyi associated with pathogenicity in tiger shrimp (Penaeus monodon) larvae. Five luminescent and four nonluminescent V. harveyi isolates were subjected to phenotyping and random amplified polymorphic DNA (RAPD) fingerprinting, and pathogenicity testing to P. monodon mysis. Four isolates induced 34-41% mortality of P. monodon mysis when challenged at the rate of 10(6) CFU ml(-1) within 60 h. Sucrose-fermenting biotypes of V. harveyi appeared to be associated with pathogenicity to larval shrimp. Higher temperature and salinity appeared to play a role on the onset of vibriosis and mortality in the challenged larval shrimp. Pathogenic isolates of V. harveyi could be demarcated as revealed by their clustering in the dendrogram constructed based on the RAPD fingerprints. Nonluminescent V. harveyi also appear to be important aetiological agents of vibriosis of shrimp larvae. Sucrose-fermenting biotypes are likely to be pathogenic. High temperature may trigger onset of vibriosis. Biotyping of V. harveyi isolates and looking for traits, such as ability to ferment sucrose may be helpful in identifying the pathogenic forms, and such approach requires to be investigated further with larger number of isolates.

  14. Pathogenicity Assay of Vibrio harveyi in Tiger Shrimp Larvae Employing Rifampicin-Resistant as A Molecular Marker

    OpenAIRE

    . Widanarni; D. Meha; Sri Nuryati; . Sukenda; A. Suwanto

    2007-01-01

    Rifampicin-resistant marker was employed as a reporter to assay pathogenicity of Vibrio harveyi  in shrimp larvae.  V. harveyi M. G3 and G7 that difference not schizotyping as shown by Pulsed-Filed Gel Electrophoresis (PFGE) used in this study. Spontaneous mutation was conducted to generate V. harveyi resistant to rifampicin. Two groups of shrimp post-larvae (PL5) were immersed for 30 min in 106 CFU/ml of mutants and wild type of V. harveyi, respectively; and then placed in a 2 liter shrimp r...

  15. Efficacy of potential phage cocktails against Vibrio harveyi and closely related Vibrio species isolated from shrimp aquaculture environment in the south east coast of India.

    Science.gov (United States)

    Stalin, Nattan; Srinivasan, Pappu

    2017-08-01

    A diverse set of novel phages infecting the marine pathogenic Vibrio harveyi was isolated from shrimp aquaculture environments in the south east coast of India. Based on initial screening, three phages with a broad host range revealed that the growth inhibition of phage is relatively specific to V. harveyi. They were also able to infect V. alginolyticus and V. parahemolyticus that belonged to the Harveyi clade species from shrimp pond and sea coast environment samples. However, the impact of these phages on their host bacterium are well understood; a one-step growth curve experiment and transmission electron microscope (TEM) revealed three phages grouped under the Myoviridae (VHM1 and VHM2); Siphoviridae (VHS1) family. These phages were further molecular characterized with respect to phage genomic DNA isolates. The randomly amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) digestion with HindIII, and major structural proteins were distinguished by sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) clearly indicated that all the phage isolates were different, even when they came from the same source, giving an insight into the diversity of phages. Evaluation of microcosm studies of Penaeus monodon larvae infected with V. harveyi (105 CFU mL-1) showed that larvae survival after 96 h in the presence of phage treatment at 109 PFU mL-1 was enhanced when compared with the control. The resolution in over survival highly recommended that this study provides the phage-based therapy which could be an innovative and eco-friendly solution against Vibrio disease in shrimp aquaculture and in the natural environment. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Adherence And Pathogenicity Assay Of Vibrio Harveyi In Tiger Shrimp (Penaeus Monodon) Larvae For Screening Biocontrol Agent

    OpenAIRE

    SUWANTO, ANTONIUS; HALA, YUSMINAH; AFFANDI, RIDWAN; Jr., MUHAMMAD ZAIRIN

    2002-01-01

    Rifampicin-resistant marker was employed as a reporter to detect the adherence and colonization of V. harveyi in shrimp larvae. Vibrio harveyi P1B and YA32.2 were isolated from dead shrimp larvae in Besuki, Northern Coast of East Java, while V. harveyi HB3, was isolated from pristine sea water in Pacitan, Southern Coast of East Java. Vibrio metschnikovii used as biocontrol agent was isolated from healthy shrimp larvae in Serang, West Java. Spontaneous mutation was conducted to generate V. h...

  17. Comparison of Vibrio harveyi strains isolated from shrimp farms and from culture collection in terms of toxicity and antibiotic resistance.

    Science.gov (United States)

    Nakayama, Tatsuya; Ito, Emi; Nomura, Nakao; Nomura, Nobuhiko; Matsumura, Masatoshi

    2006-05-01

    Vibrio harveyi strains isolated from shrimp farms (wild strains) were compared with those from culture collections in terms of minimum inhibitory concentration (MIC) and toxicity. Wild strains had higher MIC values for four antibiotics (kanamycin, carbenicillin, oxytetracycline and ampicillin) and also showed higher toxicity compared with culture collection strains. Vibrio harveyi with the lowest antibacterial resistance was chosen to test if a gradual increase in antibiotic concentration and frequent subculture would enhance its antibiotic resistance. Results showed that V. harveyi was able to develop resistance to oxytetracycline. The MIC value was 250 times higher compared with the MIC before subculturing. Moreover, the V. harveyi strain developed slightly higher toxicity. Therefore, it is possible that there is a relationship between antibiotic resistance and toxicity in V. harveyi.

  18. A soluble fatty acyl-acyl carrier protein synthetase from the bioluminescent bacterium Vibrio harveyi.

    Science.gov (United States)

    Byers, D M; Holmes, C G

    1990-01-01

    An enzyme catalyzing the ligation of long chain fatty acids to bacterial acyl carrier protein (ACP) has been detected and partially characterized in cell extracts of the bioluminescent bacterium Vibrio harveyi. Acyl-ACP synthetase activity (optimal pH 7.5-8.0) required millimolar concentrations of ATP and Mg2+ and was slightly activated by Ca2+, but was inhibited at high ionic strength and by Triton X-100. ACP from either Escherichia coli (apparent Km = 20 microM) or V. harveyi was used as a substrate. Of the [14C]fatty acids tested as substrates (8-18 carbons), a preference for fatty acids less than or equal to 14 carbons in length was observed. Vibrio harveyi acyl-ACP synthetase appears to be a soluble hydrophilic enzyme on the basis of subcellular fractionation and Triton X-114 phase partition assay. The enzyme was not coinduced with luciferase activity or light emission in vivo during the late exponential growth phase in liquid culture. Acyl-ACP synthetase activity was also detected in extracts from the luminescent bacterium Vibrio fischeri, but not Photobacterium phosphoreum. The cytosolic nature and enzymatic properties of V. harveyi acyl-ACP synthetase indicate that it may have a different physiological role than the membrane-bound activity of E. coli, which has been implicated in phosphatidylethanolamine turnover. Acyl-ACP synthetase activity in V. harveyi could be involved in the intracellular activation and elongation of exogenous fatty acids that occurs in this species or in the reactivation of free myristic acid generated by luciferase.

  19. 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.

  20. Isolation and characterization of exopolysaccharide produced by Vibrio harveyi strain VB23.

    Science.gov (United States)

    Bramhachari, P V; Dubey, S K

    2006-11-01

    The aim of the study was to isolate and characterize exopolysaccharide (EPS) produced by Vibrio harveyi strain VB23. Growth and EPS production by V. harveyi strain VB23, was studied in mineral salts medium supplemented with NaCl (1.5%) and glucose (0.2%). The rate of EPS production in batch cultures was highest during the late log phase of growth when compared with stationary growth phase. The exopolymer was recovered from the culture supernatant by using a cold ethanol precipitation-dialysis procedure. Chemical analyses of EPS revealed that it is primarily composed of neutral sugars, uronic acids, proteins and sulfates. The purified EPS revealed prominent functional reactive groups, such as hydroxyl, carboxylic and amides, which correspond to a typical heteropolymeric polysaccharide and the EPS, also possessed good emulsification activity. The gas chromatographic analysis of an alditol acetate-derivatized sample of EPS revealed that it is composed primarily of galactose and glucose. Minor components found were rhamnose, fucose, ribose, arabinose, xylose and mannose. The EPS produced by V. harveyi strain VB23 is a heteropolysaccharide possessing good emulsification activity. EPS was readily isolated from culture supernatants, which suggests that the EPS was a slime-like EPS. This is the first report of EPS characterization in luminous V. harveyi bacteria, which describes the isolation and characterization of an EPS expressed by V. harveyi. The results of the study contributes significantly towards an understanding of the chemical composition and applications of the EPS in environmental biotechnology and bioremediation.

  1. Transferable chloramphenicol resistance determinant in luminous Vibrio harveyi from penaeid shrimp Penaeus monodon larvae

    Directory of Open Access Journals (Sweden)

    Thangapalam Jawahar Abraham

    2016-12-01

    Full Text Available Antibiotic-resistant luminous Vibrio harveyi strains isolated from Penaeus monodon larvae were screened for the possession of transferable resistance determinants. All the strains were resistant to chloramphenicol and the determinant coding for chloramphenicol resistance was transferred to Escherichia coli at frequencies of 9.50x10-4 to 4.20x10-4. The results probably suggest the excessive use of chloramphenicol in shrimp hatcheries to combat luminous vibriosis.

  2. Monitoring of Vibrio harveyi quorum sensing activity in real time during infection of brine shrimp larvae.

    Science.gov (United States)

    Defoirdt, Tom; Sorgeloos, Patrick

    2012-12-01

    Quorum sensing, bacterial cell-to-cell communication, has been linked to the virulence of pathogenic bacteria. Indeed, in vitro experiments have shown that many bacterial pathogens regulate the expression of virulence genes by this cell-to-cell communication process. Moreover, signal molecules have been detected in samples retrieved from infected hosts and quorum sensing disruption has been reported to result in reduced virulence in different host-pathogen systems. However, data on in vivo quorum sensing activity of pathogens during infection of a host are currently lacking. We previously reported that quorum sensing regulates the virulence of Vibrio harveyi in a standardised model system with gnotobiotic brine shrimp (Artemia franciscana) larvae. Here, we monitored quorum sensing activity in Vibrio harveyi during infection of the shrimp, using bioluminescence as a read-out. We found that wild-type Vibrio harveyi shows a strong increase in quorum sensing activity early during infection. In this respect, the bacteria behave remarkably similar in different larvae, despite the fact that only half of them survive the infection. Interestingly, when expressed per bacterial cell, Vibrio harveyi showed around 200-fold higher maximal quorum sensing-regulated bioluminescence when associated with larvae than in the culture water. Finally, the in vivo quorum sensing activity of mutants defective in the production of one of the three signal molecules is consistent with their virulence, with no detectable in vivo quorum sensing activity in AI-2- and CAI-1-deficient mutants. These results indicate that AI-2 and CAI-1 are the dominant signals during infection of brine shrimp.

  3. Shrimp pathogenicity, hemolysis, and the presence of hemolysin and TTSS genes in Vibrio harveyi isolated from Thailand.

    Science.gov (United States)

    Rattanama, Pimonsri; Srinitiwarawong, Kanchana; Thompson, Janelle R; Pomwised, Rattanaruji; Supamattaya, Kidchakarn; Vuddhakul, Varaporn

    2009-09-23

    The virulence factors of Vibrio harveyi, the causative agent of luminous vibriosis, are not completely understood. We investigated the correlations between shrimp mortality, hemolysis, the presence of a hemolysin gene (vhh), and a gene involved in the type III secretion system (the Vibrio calcium response gene vcrD). V harveyi HY01 was isolated from a shrimp that died from vibriosis, and 36 other V. harveyi isolates were obtained from fish and shellfish in Hat Yai city, Thailand. An ocean isolate of V. harveyi BAA-1116 was also included. Thirteen isolates including V harveyi HYO1 caused shrimp death 12 h after injection. Most V harveyi isolates in this group (designated as Group A) caused hemolysis on prawn blood agar. None of the shrimp died after injection with V harveyi BAA-1116. Molecular analysis of all V harveyi isolates revealed the presence of vcrD in both pathogenic and non-pathogenic strains. Although vhh was detected in all V harveyi isolates, some isolates did not cause hemolysis, indicating that vhh gene expression might be regulated. Analysis of the V harveyi HYO1 genome revealed a V cholerae like-hemolysin gene, hlyA (designated as hhl). Specific primers designed for hhl detected this gene in 3 additional V harveyi isolates but the presence of this gene was not correlated with pathogenicity. Random amplified polymorphic DNA (RAPD) analysis revealed a high degree of genetic diversity in all V harveyi isolates, and there were no correlations among the hhl-positive isolates or the pathogenic strains.

  4. Passive Immune-Protection of Litopenaeus vannamei against Vibrio harveyi and Vibrio parahaemolyticus Infections with Anti-Vibrio Egg Yolk (IgY-Encapsulated Feed

    Directory of Open Access Journals (Sweden)

    Xiaojian Gao

    2016-05-01

    Full Text Available Vibrio spp. are major causes of mortality in white shrimp (Litopenaeus vannamei which is lacking adaptive immunity. Passive immunization with a specific egg yolk antibody (IgY is a potential method for the protection of shrimp against vibriosis. In this study, immune effects of the specific egg yolk powders (IgY against both V. harveyi and V. parahaemolyticus on white shrimp were evaluated. The egg yolk powders against V. harveyi and V. parahaemolyticus for passive immunization of white shrimp were prepared, while a tube agglutination assay and an indirect enzyme-linked immunosorbent assay (ELISA were used for detection of IgY titer. Anti-Vibrio egg yolk was encapsulated by β-cyclodextrin, which could keep the activity of the antibody in the gastrointestinal tract of shrimp. The results showed that the anti-Vibrio egg powders had an inhibiting effect on V. harveyi and V. parahaemolyticus in vitro. Lower mortality of infected zoeae, mysis, and postlarva was observed in groups fed with anti-Vibrio egg powders, compared with those fed with normal egg powders. The bacterial load in postlarva fed with specific egg powders in seeding ponds was significantly lower than those fed with normal egg powders in seeding ponds. These results show that passive immunization by oral administration with specific egg yolk powders (IgY may provide a valuable protection of vibrio infections in white shrimp.

  5. Molecular variations in Vibrio alginolyticus and V. harveyi in shrimp-farming systems upon stress

    Directory of Open Access Journals (Sweden)

    Anix Vivek Santhyia

    2015-01-01

    Full Text Available A study was performed to investigate the genomic variations in the shrimp farm isolates of Vibrio alginolyticus and V. harveyi when the isolates were subjected to environmental stress. Samples of shrimps, water and sediment were collected from Southern Indian coastal shrimp farms. Vibrio isolates were biochemically identified and confirmed using 16S rDNA and gyrB gene specific PCR. The bacterial strains were genotyped by PCR fingerprinting using GTG(5 and IS (Insertion Sequence primers. Seven strains each of V. alginolyticus and V. harveyi were subjected to 10 passages through trypticase soya broth (TSB, which contained different NaCl concentrations (3, 6 and 8% and trypticase soya agar (TSA. V. alginolyticus was also passaged through TSB with a 12% NaCl concentration. PCR fingerprinting, which was performed on the strains that were passaged through different salt concentrations, confirmed that V. alginolyticus and V. harveyi could affect the genomic variations, depending on the environmental conditions of the culture. The study highlights the complex genotypic variations that occur in Vibrio strains of tropical aquatic environment because of varied environmental conditions, which result in genetic divergence and/or probable convergence. Such genetic divergence and/or convergence can lead to the organismal adaptive variation, which results in their ability to cause a productive infection in aquatic organisms or generation of new strains.

  6. Exposure to static magnetic field stimulates quorum sensing circuit in luminescent Vibrio strains of the Harveyi clade.

    Directory of Open Access Journals (Sweden)

    Adelfia Talà

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Byers, D.M.

    1989-01-01

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

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

    International Nuclear Information System (INIS)

    Byers, D.M.

    1989-01-01

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

  9. A simple and rapid immunochromatographic test strip for detection of pathogenic isolates of Vibrio harveyi.

    Science.gov (United States)

    Sithigorngul, Paisarn; Rukpratanporn, Sombat; Pecharaburanin, Nilawan; Suksawat, Pornthip; Longyant, Siwaporn; Chaivisuthangkura, Parin; Sithigorngul, Weerawan

    2007-12-01

    Mouse monoclonal antibodies (MAbs) and rabbit polyclonal antibody (PAb) against Vibrio harveyi were generated from immunization of mice and rabbits with highly virulent isolate of V. harveyi. Two MAbs specific to virulent isolates of V. harveyi were obtained and one of them (VH4) was selected to conjugate with colloidal gold as the detector antibody was laid on a sample pad. Rabbit polyclonal antibody was used as the capture antibody at the test line (T) and goat anti-mouse IgG antibody (GAM) was used as the capture antibody at the control line (C) of nitrocellulose strip. The ready-to-use strip was held in a plastic case and then stored in a desiccated plastic bag. A sample volume of 100 microl of bacterial suspension from various sources mixed with application buffer was applied to the sample chamber at one end of the strip and allowed to flow by chromatography through the nitrocellulose membrane to the other end. In test samples containing virulent isolates of V. harveyi, the bacteria would bind to the monoclonal antibody conjugated with colloidal gold and the resulting complex would be captured by the antibodies at the test line to give a reddish-purple band. Any unbound monoclonal antibody conjugated with colloidal gold moved across the test line would be captured by the GAM and form a band at the control line (C). In sample without V. harveyi or with V. harveyi below the limit (culture. The beneficial features of this kit are that simple, convenient and quick results (within 15 min) can be obtained without the requirement of sophisticated tools or special equipments and skills.

  10. Infection with Photobacterium damselae subspecies damselae and Vibrio harveyi in snapper, Pagrus auratus with bloat.

    Science.gov (United States)

    Stephens, F J; Raidal, S R; Buller, N; Jones, B

    2006-05-01

    To diagnose the cause of chronic, low mortality associated with bloat in tanks of snapper at an aquaculture facility. A clinical, pathological and microbiological investigation into the cause of a low number of ongoing mortalities associated with bloat in snapper at an aquaculture facility is outlined. Necropsy, histology, microbiology and a comparison of haematology and water analysis from affected and unaffected fish and holding tanks, respectively were conducted. Affected moribund fish were found in lateral or dorsal recumbency floating on the water surface within 24 hours of death. Photobacterium damselae subspecies damselae was isolated from intestinal contents and Vibrio harveyi from the blood of affected fish and both were isolated from culture water. Both V harveyi and P damselae subspecies damselae isolates were sensitive to tetracycline, ciprofloxacin and sulphamethoxazole plus trimethoprim. Environmental parameters such as pH and dissolved oxygen were similar in tanks of affected and unaffected fish. Affected fish had gas distended swimbladders, anaemia, and the intestines were diffusely distended with a clear, pale yellowish fluid. Livers were mottled tan and green in a zonal pattern. Histologically the intestines of fish from tanks suffering mortality had a moderate granulocytic enteritis with oedema and infiltrations with eosinophilic granule cells that were also present as an infiltrate in the gills. There were elevated numbers of melanomacrophage centres and haemosiderin deposits in the spleen, kidney and liver of affected fish. Vibrio harveyi and Photobacterium damselae subspecies damselae infection should be recognised as potential pathogens of snapper held in water of less than optimal quality.

  11. Extract from the fermented soybean product Natto inhibits Vibrio biofilm formation and reduces shrimp mortality from Vibrio harveyi infection.

    Science.gov (United States)

    Yatip, Pattanan; Nitin Chandra Teja, D; Flegel, Timothy W; Soowannayan, Chumporn

    2018-01-01

    Many bacteria, including Vibrio pathogens of shrimp, need to colonize and/or form biofilms in hosts or the environment to cause disease. Thus, one possible control strategy for shrimp Vibriosis is biofilm inhibition. With this objective, an extract from the Japanese fermented soybean product, Natto was tested with the luminescent shrimp pathogen Vibrio harveyi (VH) for its ability to inhibit or degrade biofilm and to interfere with cell growth in broth. Natto is a traditional fermentation product of Bacillus subtilis var Natto (BSN1). Using 96 well microtiter plates coated with 0.4% chitosan, we found that biofilm formation by VH was inhibited, while growth in parallel broth cultures was not. When an extract from Natto prepared using BSN1 was mixed with feed for the whiteleg shrimp Penaeus vannamei before immersion challenge with V. harveyi at 10 6  cfu/ml, survival was significantly higher (p≤0.05) than for control shrimp given feed without these additives. Further work done to test whether d-amino acids were involved in biofilm formation as previously reported for B. subtilis, Staphylococus aureus and Pseudomonas aeruginosa gave negative results. In conclusion, we discovered that Natto extract can inhibit Vibrio biofilm formation and that it or BSN1 alone added to shrimp feed can significantly reduce shrimp mortality in immersion challenges with pathogenic VH. This shows some promise for possible application against Vibriosis in shrimp since Natto is generally regarded as safe (GRAS) for human consumption. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Inhibitory Mechanism of Robiotic Bacteria on The Growth of Vibrio harveyi in Tiger Shrimp (Penaeus monodon Larvae

    Directory of Open Access Journals (Sweden)

    . Widanarni

    2008-07-01

    Full Text Available Three probiotics named SKT-b, 1Ub, and Ua had inhibitory activity against the growth of Vibrio harveyi. These strains were mutated by rifampicin resistant. The inhibitory effect of SKT-b,1Ub, and Ua on the growth of V. harveyi was investigated by concomitant incubation of the two bacteria in a culture shrimp larvae. Colony forming unit of V. harveyi, probiotic and total of bacteria in dead, live larvae and water culture was monitored, and survival rate of larvae was investigated. Shrimp inoculated probiotic previously had survival rate higher than control (without probiotic. Number of V. harveyi in treatment without probiotic inoculation also higher compared to treatment with probiotic inoculation in dead, live larvae and water culture.  It demonstrated possible inhibition of probiotic bacteria on V. harveyi through competition for adherence sites or nutrition source. Partial sequencing of 16S-rRNA gene showed that 1Ub was similar to Pseudoalteromonas piscicida, whereas SKT-b and Ua were similar to Vibrio alginolyticus. Keywords: probiotic bacteria, inhibitory mechanism, V. harveyi, tiger shrimp   ABSTRAK Tiga isolat bakteri probiotik yaitu 1Ub, SKT-b dan Ua telah diuji memiliki aktivitas penghambatan terhadap pertumbuhan V. harveyi secara in vitro. Ketiga isolat ini kemudian diberi penanda resisten rifampisin (RfR melalui mutasi spontan untuk mempelajari mekanisme penghambatannya pada larva udang windu.  Efek penghambatan dari 1Ub, SKT-b dan Ua terhadap pertumbuhan V. harveyi diamati melalui pemberian secara bersamaan antara bakteri probiotik dan V. harveyi tersebut dalam air pemeliharaan larva udang.  Jumlah sel bakteri probiotik, V. harveyi dan total bakteri baik pada larva mati, larva hidup dan air pemeliharaan diamati dan kelangsungan hidup larva dihitung.  Nilai kelangsungan hidup udang pada perlakuan yang diinokulasi bakteri probiotik lebih tinggi daripada kontrol (tanpa penambahan bakteri probiotik. Jumlah sel V. harveyi pada

  13. Identification of AI-2 Quorum Sensing Inhibitors in Vibrio harveyi Through Structure-Based Virtual Screening.

    Science.gov (United States)

    Jiang, Tianyu; Zhu, Peng; Du, Lupei; Li, Minyong

    2018-01-01

    Quorum sensing (QS) is a cell-to-cell communication system that regulates gene expression as a result of the production and perception of signal molecules called autoinducers (AIs). AI-2 is a QS autoinducer produced by both Gram-negative and Gram-positive bacteria, in which it regulates intraspecies and interspecies communication. The identification of QS inhibitors is considered a promising strategy for the development of anti-virulence drugs with reduced selective pressure for resistance. Here we describe a high-throughput virtual screening approach to identify AI-2 quorum sensing inhibitors on the basis of Vibrio harveyi LuxPQ crystal structure. Seven potent inhibitors with IC 50 values in the micromolar range were selected with no effect or low effect on V. harveyi growth rate.

  14. Norepinephrine and dopamine increase motility, biofilm formation and virulence of Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    Qian eYang

    2014-11-01

    Full Text Available Vibrio harveyi is one of the major pathogens of aquatic organisms, affecting both vertebrates and invertebrates, and causes important losses in the aquaculture industry. In order to develop novel methods to control disease caused by this pathogen, we need to obtain a better understanding of pathogenicity mechanisms. Sensing of catecholamines increases both growth and production of virulence-related factors in pathogens of terrestrial animals and humans. However, at this moment, knowledge on the impact of catecholamines on the virulence of pathogens of aquatic organisms is lacking. In the present study, we report that in V. harveyi, norepinephrine and dopamine increased growth in serum-supplemented medium, siderophore production, swimming motility and expression of genes involved in flagellar motility, biofilm formation, and exopolysaccharide production. Consistent with this, pretreatment of V. harveyi with catecholamines prior to inoculation into the rearing water resulted in significantly decreased survival of gnotobiotic brine shrimp larvae, when compared to larvae challenged with untreated V. harveyi. Further, norepinephrine-induced effects could be neutralized by α-adrenergic antagonists or by the bacterial catecholamine receptor antagonist LED209, but not by β-adrenergic or dopaminergic antagonists. Dopamine-induced effects could be neutralized by dopaminergic antagonists or LED209, but not by adrenergic antagonists. Together, our results indicate that catecholamine sensing increases the success of transmission of V. harveyi and that interfering with catecholamine sensing might be an interesting strategy to control vibriosis in aquaculture. We hypothesise that upon tissue and/or hemocyte damage during infection, pathogens come into contact with elevated catecholamine levels, and that this stimulates the expression of virulence factors that are required to colonize a new host.

  15. Norepinephrine and dopamine increase motility, biofilm formation, and virulence of Vibrio harveyi.

    Science.gov (United States)

    Yang, Qian; Anh, Nguyen D Q; Bossier, Peter; Defoirdt, Tom

    2014-01-01

    Vibrio harveyi is one of the major pathogens of aquatic organisms, affecting both vertebrates and invertebrates, and causes important losses in the aquaculture industry. In order to develop novel methods to control disease caused by this pathogen, we need to obtain a better understanding of pathogenicity mechanisms. Sensing of catecholamines increases both growth and production of virulence-related factors in pathogens of terrestrial animals and humans. However, at this moment, knowledge on the impact of catecholamines on the virulence of pathogens of aquatic organisms is lacking. In the present study, we report that in V. harveyi, norepinephrine (NE) and dopamine (Dopa) increased growth in serum-supplemented medium, siderophore production, swimming motility, and expression of genes involved in flagellar motility, biofilm formation, and exopolysaccharide production. Consistent with this, pretreatment of V. harveyi with catecholamines prior to inoculation into the rearing water resulted in significantly decreased survival of gnotobiotic brine shrimp larvae, when compared to larvae challenged with untreated V. harveyi. Further, NE-induced effects could be neutralized by α-adrenergic antagonists or by the bacterial catecholamine receptor antagonist LED209, but not by β-adrenergic or dopaminergic antagonists. Dopa-induced effects could be neutralized by dopaminergic antagonists or LED209, but not by adrenergic antagonists. Together, our results indicate that catecholamine sensing increases the success of transmission of V. harveyi and that interfering with catecholamine sensing might be an interesting strategy to control vibriosis in aquaculture. We hypothesize that upon tissue and/or hemocyte damage during infection, pathogens come into contact with elevated catecholamine levels, and that this stimulates the expression of virulence factors that are required to colonize a new host.

  16. Aktivitas Antibakteri Ekstrak Jeruju (Acanthus ilicifolius terhadap Pertumbuhan Vibrio harveyi Secara in vitro (ANTIBACTERIAL ACTIVITY OF JERUJU (ANACTHUS ILICIFOLIUS EXTRACTS ON THE GROWTH OF VIBRIO HARVEYI IN VITRO

    Directory of Open Access Journals (Sweden)

    Gina Saptiani

    2013-07-01

    Full Text Available The aim of this research was to study the potential of jeruju (Acanthus ilicifolius extract as anantibacterial for alternative therapy and control of bacterial diseases in prawn nurseries. Ethanol extractionwas prepared from jeruju’s leaves, trunks, fruits, and flowers..Each extract was prepared at differentconcentrations (50 ppm, 100 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm,900 ppm, and 1000 ppm, respectively and further tested its antibacterial activity against Vibrio harveyiusing the agar disc diffusion method. The results showed that A. ilicifolius is a potential antibacterial,extract of the leaves seemed to be more effective in inhibit the growth of V. harveyi compared to other partsof the plant.

  17. Chemical and structural characterization of hydroxamate siderophore produced by marine Vibrio harveyi.

    Science.gov (United States)

    Murugappan, R M; Aravinth, A; Karthikeyan, M

    2011-02-01

    In the present study, 22 different bacteria were isolated from open ocean water from the Gulf of Mannar, India. Of the 22 isolates, 4 were identified as Vibrio spp. (VM1, VM2, VM3 and VM4) and found to produce siderophores (iron-binding chelators) under iron-limited conditions. Different media were found to have an influence on siderophore production. Maximum siderophore production was observed with VM1 isolate in MM9 salts medium at 48 h of incubation. The isolate was confirmed as Vibrio harveyi based on 16S rRNA gene sequencing and phylogenetic analysis. Fourier-transform infrared (FTIR) and (1)H nuclear magnetic resonance (NMR) spectra revealed the hydroxamate nature of the siderophore produced. Further characterization of the siderophore revealed it to be of dihydroxamate nature, forming hexadentate ligands with Fe(III) ions. A narrow shift in ultraviolet (UV)-Vis spectrum was observed on photolysis due to ligand oxidation. Growth-promotion bioassay with Aeromonas hydrophila, Staphylococcus aureus and E. coli confirmed the iron-scavenging property of the siderophore produced by Vibrio harveyi.

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

    Digital Repository Service at National Institute of Oceanography (India)

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

    , direct viable counts, actively respiring cell counts, nucleoid-containing cell counts, and total counts were determined. V. harveyi incubated at 22 degrees C nutrient-limited artificial seawater (ASW) became nonculturable after approximately 62 and 45 d...

  19. 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.

  20. A novel bacteriocin-like substance (BLIS) from a pathogenic strain of Vibrio harveyi.

    Science.gov (United States)

    Prasad, Sathish; Morris, Peter C; Hansen, Rasmus; Meaden, Philip G; Austin, Brian

    2005-09-01

    Inter-strain and inter-species inhibition mediated by a bacteriocin-like inhibitory substance (BLIS) from a pathogenic Vibrio harveyi strain VIB 571 was demonstrated against four isolates of the same species, and one culture each of a Vibrio sp., Vibrio fischeri, Vibrio gazogenes and Vibrio parahaemolyticus. The crude BLIS, which was obtained by ammonium-sulphate precipitation of the cell-free supernatant of a 72 h broth culture of strain VIB 571, was inactivated by lipase, proteinase K, pepsin, trypsin, pronase E, SDS and incubation at > or =60 degrees C for 10 min. The activity was stable between pH 2-11 for at least 5 h. Anion-exchange chromatography, gel filtration, SDS-PAGE and two-dimensional gel electrophoresis revealed the presence of a single major peak, comprising a protein with a pI of approximately 5.4 and a molecular mass of approximately 32 kDa. The N-terminal amino acid sequence of the protein comprised Asp-Glu-Tyr-Ile-Ser-X-Asn-Lys-X-Ser-Ser-Ala-Asp-Ile (with X representing cysteine or modified amino acid residues). A similarity search based on the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) generated peptide masses and the N-terminal sequence did not yield any significant matches.

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

    Directory of Open Access Journals (Sweden)

    Anetzberger Claudia

    2012-09-01

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

  2. Sequence and function of LuxO, a negative regulator of luminescence in Vibrio harveyi.

    Science.gov (United States)

    Bassler, B L; Wright, M; Silverman, M R

    1994-05-01

    Density-dependent expression of luminescence in Vibrio harveyi is regulated by the concentration of extracellular signal molecules (autoinducers) in the culture medium. A recombinant clone that restored function to one class of spontaneous dim mutants was found to encode a function required for the density-dependent response. Transposon Tn5 insertions in the recombinant clone were isolated, and the mutations were transferred to the genome of V. harveyi for examination of mutant phenotypes. Expression of luminescence in V. harveyi strains with transposon insertions in one locus, luxO, was independent of the density of the culture and was similar in intensity to the maximal level observed in wild-type bacteria. Sequence analysis of luxO revealed one open reading frame that encoded a protein, LuxO, similar in amino acid sequence to the response regulator domain of the family of two-component, signal transduction proteins. The constitutive phenotype of LuxO- mutants indicates that LuxO acts negatively to control expression of luminescence, and relief of repression by LuxO in the wild type could result from interactions with other components in the Lux signalling system.

  3. Effect of combined function of temperature and water activity on the growth of Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    Kang Zhou

    2012-12-01

    Full Text Available Vibrio harveyi is considered as a causative agent of the systemic disease, vibriosis, which occurs in many biological fields. The effects of temperatures (12.9-27.1 ºC and water activity (NaCl% 0.6%-3.4% on V. harveyi were investigated. The behavior and growth characteristics of V. harveyi was studied and modeled. Growth curves were fitted by using Gompertz and Baranyi models, and the Baranyi model showed a better fittness. Then, the maximum growth rates (µmax and lag phase durations (LPD, λ obtained from both Gompertz and Baranyi model were modeled as a combination function of temperature and water activity using the response surface and Arrhenius-Davey models for secondary model. The value of r², MSE, bias and accuracy factor suggest Baranyi model has better fitness than Gompertz model. Furthermore, validation of the developed models with independent data from ComBase also shown better interrelationship between observed and predicted growth parameter when using Baranyi model.

  4. Effect of combined function of temperature and water activity on the growth of Vibrio harveyi.

    Science.gov (United States)

    Zhou, Kang; Gui, Meng; Li, Pinglan; Xing, Shaohua; Cui, Tingting; Peng, Zhaohui

    2012-10-01

    Vibrio harveyi is considered as a causative agent of the systemic disease, vibriosis, which occurs in many biological fields. The effects of temperatures (12.9-27.1 °C) and water activity (NaCl% 0.6%-3.4%) on V. harveyi were investigated. The behavior and growth characteristics of V. harveyi was studied and modeled. Growth curves were fitted by using Gompertz and Baranyi models, and the Baranyi model showed a better fittness. Then, the maximum growth rates (μmax) and lag phase durations (LPD, λ) obtained from both Gompertz and Baranyi model were modeled as a combination function of temperature and water activity using the response surface and Arrhenius-Davey models for secondary model. The value of r(2), MSE, bias and accuracy factor suggest Baranyi model has better fitness than Gompertz model. Furthermore, validation of the developed models with independent data from ComBase also shown better interrelationship between observed and predicted growth parameter when using Baranyi model.

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

    Science.gov (United States)

    Anetzberger, Claudia; Schell, Ursula; Jung, Kirsten

    2012-09-18

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

  6. Chitin Degradation Proteins Produced by the Marine Bacterium Vibrio harveyi Growing on Different Forms of Chitin.

    Science.gov (United States)

    Svitil, A L; Chadhain, S; Moore, J A; Kirchman, D L

    1997-02-01

    Relatively little is known about the number, diversity, and function of chitinases produced by bacteria, even though chitin is one of the most abundant polymers in nature. Because of the importance of chitin, especially in marine environments, we examined chitin-degrading proteins in the marine bacterium Vibrio harveyi. This bacterium had a higher growth rate and more chitinase activity when grown on (beta)-chitin (isolated from squid pen) than on (alpha)-chitin (isolated from snow crab), probably because of the more open structure of (beta)-chitin. When exposed to different types of chitin, V. harveyi excreted several chitin-degrading proteins into the culture media. Some chitinases were present with all of the tested chitins, while others were unique to a particular chitin. We cloned and identified six separate chitinase genes from V. harveyi. These chitinases appear to be unique based on DNA restriction patterns, immunological data, and enzyme activity. This marine bacterium and probably others appear to synthesize separate chitinases for efficient utilization of different forms of chitin and chitin by-products.

  7. In vitro antibiofilm efficacy of Piper betle against quorum sensing mediated biofilm formation of luminescent Vibrio harveyi.

    Science.gov (United States)

    Srinivasan, Ramanathan; Santhakumari, Sivasubramanian; Ravi, Arumugam Veera

    2017-09-01

    Vibrio harveyi is a potent biofilm former, which confers resistance to multiple antimicrobials, disinfectants, chemicals and biocides. The prevalence of biofilm mediated antibiotic resistance among aquatic bacterial pathogens stresses the search for novel alternative approach to treat vibriosis in aquaculture. Exploring suitable therapeutics from natural resources could be a novel area of research. Therefore, this work was executed to evaluate the inhibitory effect of Piper betle ethyl acetate extract (PBE) on bioluminescence production and biofilm formation of V. harveyi. Minimal inhibitory concentration (MIC) of PBE against planktonic V. harveyi was found to be 1600 μg ml -1 ; furthermore, PBE inhibited the quorum sensing (QS) mediated bioluminescence production and biofilm formation in V. harveyi upto 98 and 74% respectively, at its sub-MIC concentration of 400 μg ml -1 without affecting their cell viability. Similar results were obtained for exopolysaccharides production and swimming motility related to biofilm formation of V. harveyi, where PBE reduced EPS production upto 64%. Light and confocal laser scanning microscopic analyses further confirmed that the PBE effectively prevented the initial attachment as well as microcolonies formation of V. harveyi biofilm, when compared to their untreated controls. This study demonstrates the promising antibiofilm activity of PBE and confirms the ethnopharmacological potential of this plant against V. harveyi infections. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Production and characterization of polyhydroxybutyrate from Vibrio harveyi MCCB 284 utilizing glycerol as carbon source.

    Science.gov (United States)

    Mohandas, S P; Balan, L; Lekshmi, N; Cubelio, S S; Philip, R; Bright Singh, I S

    2017-03-01

    Production and characterization of polyhydroxybutyrate (PHB) from moderately halophilic bacterium Vibrio harveyi MCCB 284 isolated from tunicate Phallusia nigra. Twenty-five bacterial isolates were obtained from tunicate samples and three among them exhibited an orange fluorescence in Nile red staining indicating the presence of PHB. One of the isolates, MCCB 284, which showed rapid growth and good polymer yield, was identified as V. harveyi. The optimum conditions of the isolate for the PHB production were pH 8·0, sodium chloride concentration 20 g l -1 , inoculum size 0·5% (v/v), glycerol 20 g l -1 and 72 h of incubation at 30°C. Cell dry weight (CDW) of 3·2 g l -1 , PHB content of 2·3 g l -1 and final PHB yield of 1·2 g l -1 were achieved. The extracted PHB was characterized by FTIR, NMR and DSC-TGA techniques. An isolate of V. harveyi that could effectively utilize glycerol for growth and PHB accumulation was obtained from tunicate P. nigra. PHB produced was up to 72% based on CDW. This is the first report of an isolate of V. harveyi which utilizes glycerol as the sole carbon source for PHB production with high biomass yield. This isolate could be of use as candidate species for commercial PHB production using glycerol as the feed stock or as source of genes for recombinant PHB production or for synthetic biology. © 2016 The Society for Applied Microbiology.

  9. Identification of a locus controlling expression of luminescence genes in Vibrio harveyi.

    Science.gov (United States)

    Martin, M; Showalter, R; Silverman, M

    1989-05-01

    Mutagenesis with transposon mini-Mulac was used to identify loci containing genes for bioluminescence (lux) in the marine bacterium Vibrio harveyi. Transposon insertions which resulted in a Lux- phenotype were mapped to two unlinked regions of the genome. Region I contained the luxCDABE operon which was previously shown to encode the enzymes luciferase and fatty acid reductase, which are required for light production. The other locus, region II, which was identified for the first time in this study, appeared to have a regulatory function. In Northern blot analysis of mRNA from mutants with defects in this region, no transcription from the luxCDABE operon could be detected. Strains with transposon-generated lux::lacZ gene fusions were used to analyze control of the transcription of these regions. Expression of luminescence in the wild type was strongly influenced by the density of the culture, and in strains with the lacZ indicator gene coupled to the luxCDABE operon, beta-galactosidase synthesis was density dependent. So, transcription of this operon is responsive to a density-sensing mechanism. However, beta-galactosidase synthesis in strains with lacZ fused to the region II transcriptional unit did not respond to cell density. The organization and regulation of the lux genes of V. harveyi are discussed, particularly with regard to the contrasts observed with the lux system of the fish light-organ symbiont Vibrio fischeri.

  10. Vibrio harveyi adheres to and penetrates tissues of the European abalone Haliotis tuberculata within the first hours of contact.

    Science.gov (United States)

    Cardinaud, Marion; Barbou, Annaïck; Capitaine, Carole; Bidault, Adeline; Dujon, Antoine Marie; Moraga, Dario; Paillard, Christine

    2014-10-01

    Vibrio harveyi is a marine bacterial pathogen responsible for episodic epidemics generally associated with massive mortalities in many marine organisms, including the European abalone Haliotis tuberculata. The aim of this study was to identify the portal of entry and the dynamics of infection of V. harveyi in the European abalone. The results indicate that the duration of contact between V. harveyi and the European abalone influences the mortality rate and precocity. Immediately after contact, the epithelial and mucosal area situated between the gills and the hypobranchial gland was colonized by V. harveyi. Real-time PCR analyses and culture quantification of a green fluorescent protein-tagged strain of V. harveyi in abalone tissues revealed a high density of bacteria adhering to and then penetrating the whole gill-hypobranchial gland tissue after 1 h of contact. V. harveyi was also detected in the hemolymph of a significant number of European abalones after 3 h of contact. In conclusion, this article shows that a TaqMan real-time PCR assay is a powerful and useful technique for the detection of a marine pathogen such as V. harveyi in mollusk tissue and for the study of its infection dynamics. Thus, we have revealed that the adhesion and then the penetration of V. harveyi in European abalone organs begin in the first hours of contact. We also hypothesize that the portal of entry of V. harveyi in the European abalone is the area situated between the gills and the hypobranchial gland. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  11. Isolation and characterization of Pseudoalteromonas sp. from fermented Korean food, as an antagonist to Vibrio harveyi.

    Science.gov (United States)

    Morya, V K; Choi, Wooyoung; Kim, Eun-Ki

    2014-02-01

    The microbial intervention for sustainable management of aquaculture, especially use of probiotics, is one of the most popular and practical approaches towards controlling pathogens. Vibrio harveyi is a well-known pathogenic bacterium, which is associated to a huge economic loss in the aquaculture system by causing vibriosis. The present study is crafted for screening and characterization of anti-Vibrio strains, which were isolated from various traditional fermented Korean foods. A total of 196 strains have been isolated from soybean paste (78 strains), red chili paste (49 strains), soy sauce (18 strains), jeotgal-a salted fish (34 strains), and the gazami crab-Portunus trituberculatus (17 strains). Fifteen strains showed an inhibitory effect on the growth of V. harveyi when subjected to coculture condition. Among the strains isolated, one has been identified as a significant anti-Vibrio strain. Further biochemical characterization and 16S rDNA sequencing revealed it as Pseudoalteromonas aliena, which had been deposited at the Korean Culture Center of Microorganisms (KCCM), Korea and designated as KCCM 11207P. The culture supernatants did not have any antimicrobial properties either in pure or in coculture condition. The culture supernatant was not toxic when supplemented to the swimming crab, Zoea, and Artemia larvae in aquaculture system. The results were very encouraging and showed a significant reduction in accumulated mortality. Here, we reported that pathogenic vibriosis can be controlled by Pseudoalteromonas sp. under in vitro and in vivo conditions. The results indicated that the biotic treatment offers a promising alternative to the use of antibiotics in crab aquaculture.

  12. Anaerobic respiratory growth of Vibrio harveyi, Vibrio fischeri and Photobacterium leiognathi with trimethylamine N-oxide, nitrate and fumarate: ecological implications.

    Science.gov (United States)

    Proctor, L M; Gunsalus, R P

    2000-08-01

    Two symbiotic species, Photobacterium leiognathi and Vibrio fischeri, and one non-symbiotic species, Vibrio harveyi, of the Vibrionaceae were tested for their ability to grow by anaerobic respiration on various electron acceptors, including trimethylamine N-oxide (TMAO) and dimethylsulphoxide (DMSO), compounds common in the marine environment. Each species was able to grow anaerobically with TMAO, nitrate or fumarate, but not with DMSO, as an electron acceptor. Cell growth under microaerophilic growth conditions resulted in elevated levels of TMAO reductase, nitrate reductase and fumarate reductase activity in each strain, whereas growth in the presence of the respective substrate for each enzyme further elevated enzyme activity. TMAO reductase specific activity was the highest of all the reductases. Interestingly, the bacteria-colonized light organs from the two squids, Euprymna scolopes and Euprymna morsei, and the light organ of the ponyfish, Leiognathus equus, also had high levels of TMAO reductase enzyme activity, in contrast to non-symbiotic tissues. The ability of these bacterial symbionts to support cell growth by respiration with TMAO may conceivably eliminate the competition for oxygen needed for both bioluminescence and metabolism.

  13. Inhibition of luminescence and virulence in the black tiger prawn (Penaeus monodon) pathogen Vibrio harveyi by intercellular signal antagonists.

    Science.gov (United States)

    Manefield, M; Harris, L; Rice, S A; de Nys, R; Kjelleberg, S

    2000-05-01

    Expression of luminescence in the Penaeus monodon pathogen Vibrio harveyi is regulated by an intercellular quorum sensing mechanism involving the synthesis and detection of two signaling molecules, one of which is N-hydroxy butanoyl-L-homoserine lactone and the other of which is uncharacterized. Indirect evidence has suggested that virulence, associated with a toxic extracellular protein, and luminescence in V. harveyi are coregulated. In this study the effects of an acylated homoserine lactone antagonist produced by the marine alga Delisea pulchra on luminescence and toxin production in a virulent strain of V. harveyi were analyzed. Luminescence and toxin production were both inhibited by the signal antagonist at concentrations that had no impact on growth. Toxin production was found to be prematurely induced in V. harveyi cultures incubated in a 10% conditioned medium. Additionally, a significant reduction in the toxicity of concentrated supernatant extracts from V. harveyi cultures incubated in the presence of the signal antagonist, as measured by in vivo toxicity assays in mice and prawns, was observed. These results suggest that intercellular signaling antagonists have potential utility in the control of V. harveyi prawn infections.

  14. Inhibition of Luminescence and Virulence in the Black Tiger Prawn (Penaeus monodon) Pathogen Vibrio harveyi by Intercellular Signal Antagonists

    Science.gov (United States)

    Manefield, Michael; Harris, Lachlan; Rice, Scott A.; de Nys, Rocky; Kjelleberg, Staffan

    2000-01-01

    Expression of luminescence in the Penaeus monodon pathogen Vibrio harveyi is regulated by an intercellular quorum sensing mechanism involving the synthesis and detection of two signaling molecules, one of which is N-hydroxy butanoyl-l-homoserine lactone and the other of which is uncharacterized. Indirect evidence has suggested that virulence, associated with a toxic extracellular protein, and luminescence in V. harveyi are coregulated. In this study the effects of an acylated homoserine lactone antagonist produced by the marine alga Delisea pulchra on luminescence and toxin production in a virulent strain of V. harveyi were analyzed. Luminescence and toxin production were both inhibited by the signal antagonist at concentrations that had no impact on growth. Toxin production was found to be prematurely induced in V. harveyi cultures incubated in a 10% conditioned medium. Additionally, a significant reduction in the toxicity of concentrated supernatant extracts from V. harveyi cultures incubated in the presence of the signal antagonist, as measured by in vivo toxicity assays in mice and prawns, was observed. These results suggest that intercellular signaling antagonists have potential utility in the control of V. harveyi prawn infections. PMID:10788385

  15. MARINE SPONGE Aaptos suberitoides, IT’S POTENTIAL SOURCE OF NATURAL ANTIBACTERIAL FOR CONTROLLING Vibrio harveyi ON TIGER SHRIMP (Penaeus monodon) CULTURE

    OpenAIRE

    Rosmiati Rosmiati; Andi Parenrengi; Emma Suryati

    2015-01-01

    The study aims to isolate and identify the natural antibacterial compounds potential from Aaptos suberitoides for Vibrio harveyi control on tiger shrimp (Penaeus monodon) culture. The agar diffusion method using paper discs was used to determine the antibacterial activity of extracts (diethyl ether (DEE), butanol (BUE) and aqueous (HOE) and compounds successfully isolated against Vibrio harveyi. Findings showed that the antibacterial activity was concentrated in BUE with the inhibition zone o...

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

    Science.gov (United States)

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

    2007-01-01

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

  17. Efecto de Vibrio harveyi en la sobre vivencia de larvas de Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Gabriel Aguirre-Guzmán

    2013-01-01

    Full Text Available Los cultivos acuícolas son de gran relevancia en la alimentación humana, pero crean un medio ambiente artificial que promueve el crecimiento de diferentes especies de bacterias. Las especies del género Vibrio son bacterias de la microflora normal de los camarones peneidos y son también agentes patógenos oportunistas que pueden tomar ventaja de éstos cambios ecológicos generados en los cultivos acuícolas causando diferentes enfermedades, sobrevivencias bajas y pérdidas económicas en la producción de camarón. El objetivo de la presente investigación fue determinar la variación en la sensibilidad, de diferentes subestadios larvarios (nauplio, zoea I-III, misis I-III y en el de postlarva 1, del camarón blanco del Pacifico Litopenaeus vannamei al ser expuestos a tres dosis [103, 105, y 107 unidades formadoras de colonias (UFC ml-1] de V. harveyi, mediante infecciones por inmersión (30 min. Esta especie generó una sobrevivencia baja significativa en las larvas (p < 0,05 solamente en las dosis más altas (105 y 107 UFC ml-1, siendo la última dosis la que presentó los valores más bajos de sobrevivencia. Además, se observó que los subestadios larvales y en el de postlarva 1 fueron más resistentes a V. harveyi al aumentar la edad de los mismos. Esta información tiene gran significado para la industria acuícola, ya que les permite generar estrategias que disminuyan los efectos de V. harveyi y que les permita mejorar el crecimiento y sobrevivencia de las larvas y de postlarva 1 del camarón.

  18. ANALYSIS OF IMMUNE RESPONSES ON TRANSGENIC TIGER SHRIMP (Penaeus monodon AGAINST PATHOGENIC BACTERIUM Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    Andi Parenrengi

    2014-06-01

    Full Text Available Vibriosis is one of main diseases of the black tiger shrimp Penaeus monodon infected by pathogenic bioluminous bacterium Vibrio harveyi that can cause mass mortalities in shrimp culture. The bacteria can also trigger the disease white spot syndrome virus (WSSV. An effort to produce shrimp disease-resistant strains has been done through transgenesis technology with antiviral gene transfection. By this technology, it is expected an increase in the immune response of shrimp in a variety of diseasecausing pathogens. This study aimed to determine the immune responses (total haemocytes, haemocyte differentiation, and phenoloxydase activity of transgenic tiger shrimp against pathogenic bacterium V. harveyi. Research using completely randomized design, which consists of two treatments and three replications. Test animals being used were transgenic and non-transgenic shrimp with size, weight 3.93±1.25 g and a total length of 7.59±0.87 cm. Treatments being tested were the injection of bacterium V. harveyi (density of 5x106 cfu/mL of 0.1 mL/individual on transgenic (A and non-transgenic shrimp (B. Immune response parameters such as total haemocytes, haemocyte differentiation, and phenoloxydase activity were observed on day 1, 3, and 6 days after challenging. Data were analyzed using t-test by SPSS software. The results showed that the total haemocyte of transgenic shrimp was not significantly different (P>0.05 from non-transgenic shrimp, but haemocyte differentiation and phenoloxydase activity were significantly different (P<0.05 especially on sixth days after being exposed to the bioluminescent bacteria. The study results implied that transgenic shrimp has a better immune response compared than non-transgenic shrimp.

  19. Identification and characterization of Vibrio harveyi associated with diseased abalone Haliotis diversicolor.

    Science.gov (United States)

    Jiang, Qingru; Shi, Liuyang; Ke, Caihuan; You, Weiwei; Zhao, Jing

    2013-03-26

    Mass mortality of farmed small abalone Haliotis diversicolor occurred in Fujian, China, from 2009 to 2011. Among isolates obtained from moribund abalones, the dominant species AP37 exhibited the strongest virulence. After immersion challenge with 106 CFU ml-1 of AP37, abalone mortalities of 0, 53 and 67% were induced at water temperatures of 20°C, 24°C, and 28°C, respectively. Following intramuscular injection, AP37 showed a low LD50 (median lethal concentration) value of 2.9 × 102 CFU g-1 (colony forming units per gram abalone wet body weight). The LT50 (median lethal time) values were 5.2 h for 1 × 106 CFU abalone-1, 8.4 h for 1 × 105 CFU abalone-1, and 21.5 h for 1 × 104 CFU abalone-1. For further analysis of virulence, AP37 was screened for the production of extracellular factors. The results showed that various factors including presence of flagella and production of extracellular enzymes, such as lipase, phospholipase and haemolysin, could be responsible for pathogenesis. Based on its 16S rRNA gene sequence, strain AP37 showed >98.8% similarity to Vibrio harveyi, V. campbellii, V. parahaemolyticus, V. alginolyticus, V. natriegens and V. rotiferianus, so it could not be identified by this method. However, multi-locus sequence analysis (MLSA) of concatenated sequences, including the rpoD, rctB, gyrB, toxR and pyrH genes, identified strain AP37 as V. harveyi. Phenotypic characters of AP37 were identified by API 20E. In antibiotic susceptibility tests, strain AP37 exhibited susceptibility to 7 antibiotics and resistance to 13. This is the first report of a V. harveyi-related species being linked with the mass mortality of adult abalone H. diversicolor in southern China.

  20. Nitric oxide as an antimicrobial molecule against Vibrio harveyi infection in the hepatopancreas of Pacific white shrimp, Litopenaeus vannamei.

    Science.gov (United States)

    Chen, Ting; Wong, Nai-Kei; Jiang, Xiao; Luo, Xing; Zhang, Lvping; Yang, Dan; Ren, Chunhua; Hu, Chaoqun

    2015-01-01

    Nitric oxide (NO) is a key effector molecule produced in the innate immune systems of many species for antimicrobial defense. However, how NO production is regulated during bacterial infection in invertebrates, especially crustaceans, remains poorly understood. Vibrio harveyi, a Gram-negative marine pathogen, is among the most prevalent and serious threats to the world's shrimp culture industry. Its virulence typically manifests itself through shrimp hepatopancreas destruction. In the current study, we found that NO generated by an in vitro donor system (NOC-18) could rapidly and effectively kill V. harveyi. In addition, injection of heat-killed V. harveyi increased the concentration of NO/nitrite and the mRNA expression of nitric oxide synthase (NOS) in the hepatopancreas of Pacific white shrimp (Litopenaeus vannamei), the commercially most significant shrimp species. Live V. harveyi challenge also induced NO/nitrite production and NOS gene expression in primary L. vannamei hepatopancreatic cells in a time- and dose-dependent manner. Co-incubation of l-NAME, an inhibitor selective for mammalian constitutive NOSs, dose-dependently blocked V. harveyi-induced NO/nitrite production, without affecting V. harveyi-induced NOS mRNA expression. Furthermore, l-NAME treatment significantly increased the survival rate of infecting V. harveyi in cultured primary hepatopancreatic cells of L. vannamei. As a whole, we have demonstrated that endogenous NO produced by L. vannamei hepatopancreatic cells occurs in enzymatically regulated manners and is sufficient to act as a bactericidal molecule for V. harveyi clearance. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. 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.

  2. Unstable lysogeny and pseudolysogeny in Vibrio harveyi siphovirus-like phage 1.

    Science.gov (United States)

    Khemayan, Krit; Pasharawipas, Tirasak; Puiprom, Orapim; Sriurairatana, Siriporn; Suthienkul, Orasa; Flegel, Timothy W

    2006-02-01

    Exposure of Vibrio harveyi (strain VH1114) to V. harveyi siphovirus-like phage 1 (VHS1) resulted in the production of a low percentage of lysogenized clones of variable stability. These were retrieved most easily as small colonies within dot plaques. Analysis revealed that VHS1 prophage was most likely carried by VH1114 as an episome rather than integrated into the host chromosome. In the late exponential growth phase, lysogenized VH1114 continuously produced VHS1 but also gave rise to a large number of cured progeny. The absence of phage DNA in the cured progeny was confirmed by the absence of VHS1 DNA in Southern blot and PCR assays. Curiously, these very stable, cured subclones did not show the parental phenotype of clear plaques with VHS1 but instead showed turbid plaques, both in overlaid lawns and in dot plaque assays. This phenotypic difference from the original parental isolate suggested that transient lysogeny by VHS1 had resulted in a stable genetic change in the cured clones. Such clones may be called pseudolysogens (i.e., false lysogens), since they have undergone transient lysogeny and have retained some resistance to full lytic phage development, despite the loss of viable or detectable prophage.

  3. Crustin protein Amk1 from black tiger shrimp (Penaeus monodon inhibits Vibrio harveyi and Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Moltira Tonganunt1*

    2008-05-01

    Full Text Available A crustin gene (Amk1 was identified from a haemocyte library of the black tiger shrimp, Penaeus monodon. The full-length cDNA consists of 411 bp encoding a deduced precursor of 136 amino acids with a signal peptide of 17 aminoacids. Amk1 contains a hydrophobic and a Gly-rich region at the N-terminus and a 12 conserved cysteine domain (6-DSC at the C-terminus. Transcripts of Amk1 are mainly detected in haemocytes and gills by RT-PCR analysis. A recombinant Amk1was overexpressed and purified from Escherichia coli. This has a molecular mass of 43.66 kDa with a predicted pI of 8.23. Antibacterial assays demonstrated that recombinant Amk1 exhibited antibacterial activity against Gram-positive and Gramnegativebacteria with strong inhibition against Staphylococcus aureus and Vibrio harveyi.

  4. Protective Efficacy of a Pseudoalteromonas Strain in European Abalone, Haliotis tuberculata, Infected with Vibrio harveyi ORM4.

    Science.gov (United States)

    Offret, Clément; Rochard, Vincent; Laguerre, Hélène; Mounier, Jérôme; Huchette, Sylvain; Brillet, Benjamin; Le Chevalier, Patrick; Fleury, Yannick

    2018-02-06

    The hemolymph of healthy marine invertebrates is known to harbor antibiotic-producing bacteria belonging to the genus Pseudoalteromonas. Such strains are potential probiotics to control infectious diseases in aquaculture. In the present study, we screened a collection of Pseudoalteromonas strains isolated from the hemolymph of oyster and mussel for antimicrobial activity against Vibrio harveyi, a pathogenic species responsible for high mortality in abalone. Subsequently, the protective efficacy of the most active strain named hCg-6 was investigated in abalone culture faced with a Vibrio harveyi ORM4 infection. First, we have controlled the Pseudoalteromonas hCg-6 safety for abalone health. To that end, animals were immersed for 4 h in Pseudoalteromonas hCg-6 suspensions in seawater. The abalone viability was monitored and Pseudoalteromonas hCg-6 was tracked by quantitative-PCR in abalone hemolymph. After immersion, no abalone death occurred while the strain hCg-6 was significantly detected in hemolymph. Therefore, the strain hCg-6 was considered safe for abalone and evaluated for its ability to protect abalone against V. harveyi (injection of 1 × 10 3 Vibrio per animal). A 4-h long immersion of abalone in a seawater suspension of Pseudoalteromonas hCg-6 (1 × 10 6  CFU mL -1 ) prior to infection with Vibrio harveyi significantly improved the abalone viability. Indeed, 15 days post infection, the hCg-6 treatment used increased the abalone survival rate from 16% in untreated animals to 40% in treated abalone. We hypothesized that Pseudoalteromonas hCg-6 antibacterial activity increased the hemomicrobiota shielding effect. In conclusion, Pseudoalteromonas hCg-6 is a promising anti-Vibrio strain for abalone culture.

  5. A sensitive fluorescence reporter for monitoring quorum sensing regulated protease production in Vibrio harveyi.

    Science.gov (United States)

    Rajamani, Sathish; Sayre, Richard T

    2011-02-01

    Many bacteria produce and secrete proteases during host invasion and pathogenesis. Vibrio harveyi, an opportunistic pathogen of shrimp, is known to use a two-component quorum sensing (QS) mechanism for coordination of gene expression including protease secretion at high population densities. We examined the role of V. harveyi's QS signaling molecules, N-(3-hydroxybutanoyl)-L-homoserine lactone (AI-1) and the boron derivative of autoinducer-2 (BAI-2) in extracellular protease production. A fusion protein, M3CLPY (Rajamani et al., 2007), consisting of a large protease sensitive BAI-2 mutant receptor LuxP (~38kDa) flanked by two protease insensitive cyan and yellow variants of GFP (~28kDa each) was utilized as a substrate to detect secreted protease activity. The M3CLPY fusion, with the addition of wild-type V. harveyi (BB120) cell-free culture filtrate showed a time-dependent loss in fluorescence resonance energy transfer (FRET) associated with the cleavage of the LuxP linker protein and hence separation of the two fluorophores. This cleavage of LuxP linker protein leading to decreased FRET efficiency was further confirmed by immunoblotting using anti-GFP antibody. The addition of cell-free filtrates from strains defective in one or both of the two-component QS pathways: luxN(-) (defective in AI-1), luxS(-) (defective in BAI-2), and luxN(-)/luxS(-) (defective in both AI-1/BAI-2) showed differential levels of protease production. The observed protease activities were most pronounced in wild-type, followed by the AI-1 defective mutant (BB170) and the least for luxS(-) mutant (MM30) and luxN(-)/luxS(-) double mutant (MM32) strains. Incidentally, the lowest protease producing strains MM30 and MM32 were both defective in BAI-2 production. This observation was validated by addition of synthetic BAI-2 to MM30 and MM32 strains to restore protease production. Our results indicate that BAI-2 signaling in the two-component QS pathway plays the key role in regulating

  6. Oral application of probiotic, prebiotic, and synbiotic in Pacific white shrimp(Litopenaeus vannameichallenged with Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    . Widanarni

    2013-11-01

    Full Text Available The use of antibiotics for controlling of luminous vibriosis caused by Vibrio harveyi is restricted now, because it induces antibiotic-resistant bacteria and leave residue in shrimp’s body. An alternative solution that can be done to treat the disease is by using applications of probiotic, prebiotic, and synbiotic. The aim of this research was to examine the effect of probiotic, prebiotic, and synbiotic on the survival rate and growth of Pacific white shrimp against V. harveyi infection. Feed as a treatment was supplemented with probiotic 1%, prebiotic 2%, and probiotic 1%+prebiotic 2% (synbiotic. Shrimps feed without supplementation of probiotic and prebiotic was used as a control treatment. The shrimps were maintainedin the aquarium (60×30×35 cm3 with a density of 40 shrimps/40 L and an average weight of 0.4±0.1 g. After 30 days of feeding treatment, the shrimp was challenged by immersion method with V. harveyi solution containing 106 CFU/mL. The results showed that before challenge, synbiotic feed treated shrimp has a growth rate (5.89%, feed conversionrate (1.21, and a high survival rate (80%. After challenge, survival rate (83.33% of shrimp fed diet supplemented with synbiotic was higher than prebiotic (51.67% and positive control (31.67%. Keywords: probiotic, prebiotic, synbiotic, Vibrio harveyi,Pacific white shrimp

  7. Coculture induced improved production of biosurfactant by Staphylococcus lentus SZ2: Role in protecting Artemia salina against Vibrio harveyi.

    Science.gov (United States)

    Hamza, Faseela; Kumar, Ameeta Ravi; Zinjarde, Smita

    2018-07-01

    Coculturing microorganisms can lead to enhanced production of bioactive compounds as a result of cross-species or cross-genera interactions. In this study, we demonstrate improved production of the biosurfactant (BS-SLSZ2 with antibiofilm properties) by the marine epibiotic bacterium Staphylococcus lentus SZ2 after cross-genera interactions with an aquaculture pathogen Vibrio harveyi. In cocultures, growth of V. harveyi was completely inhibited and resultant biofilms were exclusively composed of S. lentus. The cell free supernatant (CFS) derived from cocultures displayed improved antibiofilm activity with enhanced contents of BS-SLSZ2 compared to monocultured S. lentus. During coculture experiments, after short periods of incubation (6 and 12 h), 2.3 fold increased production of BS-SLSZ2 was observed. Planktonic growth of V. harveyi was also inhibited after coculturing with S. lentus as evidenced from plate culture-based studies and microscopic observations. The CFS derived from monocultures and cocultures did not display bactericidal activity and the observed inhibition of V. harveyi could be of competitive nature. During in vivo challenge experiments, S. lentus protected the model aquaculture system Artemia salina from V. harveyi infections. Seven days post infection, survival of the group of larvae infected with V. harveyi was 5 ± 4.47%. Better survival rates (73.33 ± 5.16%, comparable with the unexposed group) were observed in the group of larvae incubated with S. lentus and V. harveyi. This study highlights increased biosurfactant production by cocultured S. lentus and the application of this bacterium as a protective probiotic strain for inclusion in aquaculture practices. Copyright © 2018 Elsevier Inc. All rights reserved.

  8. The early stages of the immune response of the European abalone Haliotis tuberculata to a Vibrio harveyi infection.

    Science.gov (United States)

    Cardinaud, Marion; Dheilly, Nolwenn M; Huchette, Sylvain; Moraga, Dario; Paillard, Christine

    2015-08-01

    Vibrio harveyi is a marine bacterial pathogen responsible for episodic abalone mortalities in France, Japan and Australia. In the European abalone, V. harveyi invades the circulatory system in a few hours after exposure and is lethal after 2 days of infection. In this study, we investigated the responses of European abalone immune cells over the first 24 h of infection. Results revealed an initial induction of immune gene expression including Rel/NF-kB, Mpeg and Clathrin. It is rapidly followed by a significant immuno-suppression characterized by reduced cellular hemocyte parameters, immune response gene expressions and enzymatic activities. Interestingly, Ferritin was overexpressed after 24 h of infection suggesting that abalone attempt to counter V. harveyi infection using soluble effectors. Immune function alteration was positively correlated with V. harveyi concentration. This study provides the evidence that V. harveyi has a hemolytic activity and an immuno-suppressive effect in the European abalone. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. The Use of Synbiotics to Prevent IMNV and Vibrio harveyi Co-Infection in Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    ADNI OKTAVIANA

    2014-09-01

    Full Text Available This study evaluated the effects on viral immune responses and bacterial co-infection, of different feeding frequencies of a synbiotic supplemented diet given to Pacific white shrimp (L. vannamei. A synbiotic-supplemented diet was formulated from probiotic Vibrio alginolitycus SKT-bR and prebiotics from sweet potato (Ipomoea batatas L. oligosaccharide. Pacific white shrimp were fed with synbiotic diet at different frequencies, i.e. daily (P1, twice a week (P2, and once a week (P3 for a 30 day pre-challenge test. After the 30 day feeding period, the shrimps were challenged by intramuscular injection of Infectious Myonecrosis Virus (IMNV and Vibrio harveyi. The results showed that shrimp treated with a synbiotic-supplemented diet showed significantly higher growth performance than control groups (P<0.05. Shrimp treated under regime P1 showed the highest values for phenoloxidase (PO and respiratory burst (RB parameters compared to shrimp given with other treatments. Following the challenge test, higher survival rate were seen in the P1 treatment group, in comparison to the positive control, and the P1 treatment group showed the highest values in total haemocyte count (THC, PO, and RB.

  10. Antibacterial Property of a Coral-Associated Bacterium Pseudoalteromonas luteoviolacea Against Shrimp Pathogenic Vibrio harveyi (In Vitro Study)

    OpenAIRE

    OCKY KARNA RADJASA; TORBEN MARTENS; HANS-PETER GROSSART; AGUS SABDONO; MEINHARD SIMON; TONNY BACHTIAR

    2005-01-01

    A coral-associated bacterium was successfully screened for secondary metabolites production based on PCR amplification of the nonribosomal peptide synthetase gene and was identified as closely related to Pseudoalteromonas luteoviolacea based on its 16S rDNA. The bacterium was found to inhibit the growth of shrimp pathogenic bacterium tested, Vibrio harveyi. To characterize the inhibiting metabolite, a 279 bp long DNA fragment was obtained and the deduced amino acid sequence showed conserved s...

  11. A mathematical model and quantitative comparison of the small RNA circuit in the Vibrio harveyi and Vibrio cholerae quorum sensing systems

    International Nuclear Information System (INIS)

    Hunter, G A M; Vasquez, F Guevara; Keener, J P

    2013-01-01

    Quorum sensing is the process by which bacteria regulate their gene expression based on the local cell-population density. The quorum sensing systems of Vibrio harveyi and Vibrio cholerae are comprised of a phosphorelay cascade coupled to a small RNA (sRNA) circuit. The sRNA circuit contains multiple quorum regulated small RNA (Qrr) that regulate expression of the homologous master transcriptional regulators LuxR (in V. harveyi) and HapR (in V. cholerae). Their quorum sensing systems are topologically similar and homologous thereby making it difficult to understand why repression of HapR is more robust than LuxR to changes in Qrr. In this work we formulate and parameterize a novel mathematical model of the V. harveyi and V. cholerae sRNA circuit. We parameterize the model by fitting it to a variety of empirical data from both species. We show that we can distinguish all of the parameters and that the parameterizations (one for each species) are robust to errors in the data. We then use our model to propose some experiments to identify and explain kinetic differences between the species. We find that V. cholerae Qrr are more abundant and more sensitive to changes in LuxO than V. harveyi Qrr and argue that this is why expression of HapR is more robust than LuxR to changes in Qrr. (paper)

  12. Lactic-acid bacteria increase the survival of marine shrimp, Litopenaeus vannamei, after infection with Vibrio harveyi

    OpenAIRE

    Vieira, Felipe do Nascimento; Pedrotti, Fabiola Santiago; Buglione Neto, Celso Carlos; Mouriño, José Luiz Pedreira; Beltrame, Elpídio; Martins, Maurício Laterça; Ramirez, Cristina; Arana, Luis Alejandro Vinatea

    2007-01-01

    This study evaluated the survival, post-larvae quality, and the population of bacteria in Litopenaeus vannamei after the addition of two strains of lactic-acid bacteria (2 and B6) experimentally infected by Vibrio harveyi. Fifteen hundred nauplii were distributed in 20 L capacity tanks with four replicates. The survival of control animals was lower (21%) than that of animals fed with the strains B6 (50%) and 2 (44%). Total bacterial population in the water and larvae, as well as of the Vibrio...

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

    Science.gov (United States)

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

    2002-10-01

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

  14. Investigation of Vibrio alginolyticus, V. harveyi, and V. parahaemolyticus in large yellow croaker, Pseudosciaena crocea (Richardson reared in Xiangshan Bay, China

    Directory of Open Access Journals (Sweden)

    Lu Liu

    2016-05-01

    Full Text Available Large yellow croaker (LYC, Pseudosciaena crocea is an economically important fish species of mariculture in China. The variation of yearly production of LYC has been increasingly related to the outbreaks of fish diseases. Moreover, Vibrio infections have been identified in this fish frequently. To understand the pattern of Vibrio infections in LYC, we conducted a culture-independent survey of Vibrios in farmed LYC populations using a multiplex PCR method targeting Vibrio alginolyticus, Vibrio harveyi and Vibrio parahaemolyticus. The results showed that three fish pathogenic Vibrios had been detected in LYC populations at each sampling with a prevalence ranging from 6.7% to 73.3% but no single species dominated the Vibrio infection. The findings indicate that three Vibrio species still have impact on health status of farmed LYC and LYC aquaculture requires more efficacious prophylactic strategies. Keywords: Vibrio, Large yellow croaker, Multiplex PCR, Epidemiology

  15. Application of bacterial lipopolysaccharide to improve survival of the black tiger shrimp after Vibrio harveyi exposure.

    Science.gov (United States)

    Rungrassamee, Wanilada; Maibunkaew, Sawarot; Karoonuthaisiri, Nitsara; Jiravanichpaisal, Pikul

    2013-10-01

    This study investigates an effect of bacterial lipopolysaccharide (LPS) as feed supplement to improve immunity of the black tiger shrimp (Penaeus monodon). LPS was coated to commercial feed pellets and given to the shrimp once or twice a day for 10 days before an exposure with shrimp pathogenic bacterium Vibrio harveyi. The growth rates, percent weight gains, total hemocyte and granulocyte counts and survival rates of shrimp between the LPS-coated pellet fed groups and a control group where shrimp fed with commercial feed pellets were compared. After 10 days of the feeding trials, growth rates were not significantly different in all groups, suggesting no toxicity from LPS supplement. To determine beneficial effect of LPS diets, each group was subsequently exposed to V. harveyi by immersion method and the survival rates were recorded for seven days after the immersion. Regardless of the dosages of LPS, the shrimp groups fed with LPS-coated pellets showed higher survival rates than the control group. There was no significant difference in survival rates between the two LPS dosages groups. In addition to survival under pathogen challenge, we also determine effect of LPS on immune-related genes after 10-day feeding trial. Gene expression analysis in the P. monodon intestines revealed that antilipopolysaccharide factor isoform 3 (ALF3), C-type lectin, and mucine-like peritrophin (mucin-like PM) were expressed significantly higher in a group fed with LPS supplemental diet once or twice a day than in a control group. The transcript levels of C-type lectin and mucin-like PM had increased significantly when LPS was given once a day, while significant induction of ALF3 transcripts was observed when shrimp were fed with LPS twice a day. The up-regulation of the immune gene levels in intestines and higher resistance to V. harveyi of the shrimp fed with LPS provide the evidence for potential application of LPS as an immunostimulant in P. monodon farming. Copyright © 2013

  16. Characterization of DegQVh, a serine protease and a protective immunogen from a pathogenic Vibrio harveyi strain.

    Science.gov (United States)

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

    2008-10-01

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

  17. A nonluminescent and highly virulent Vibrio harveyi strain is associated with "bacterial white tail disease" of Litopenaeus vannamei shrimp.

    Directory of Open Access Journals (Sweden)

    Junfang Zhou

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

  18. Bacterial dynamics in intestines of the black tiger shrimp and the Pacific white shrimp during Vibrio harveyi exposure.

    Science.gov (United States)

    Rungrassamee, Wanilada; Klanchui, Amornpan; Maibunkaew, Sawarot; Karoonuthaisiri, Nitsara

    2016-01-01

    The intestinal microbiota play important roles in health of their host, contributing to maintaining the balance and resilience against pathogen. To investigate effects of pathogen to intestinal microbiota, the bacterial dynamics upon a shrimp pathogen, Vibrio harveyi, exposures were determined in two economically important shrimp species; the black tiger shrimp (BT) and the Pacific white shrimp (PW). Both shrimp species were reared under the same diet and environmental conditions. Shrimp survival rates after the V. harveyi exposure revealed that the PW shrimp had a higher resistance to the pathogen than the BT shrimp. The intestinal bacterial profiles were determined by denaturing gradient gel electrophoresis (DGGE) and barcoded pyrosequencing of the 16S rRNA sequences under no pathogen challenge control and under pathogenic V. harveyi challenge. The DGGE profiles showed that the presence of V. harveyi altered the intestinal bacterial patterns in comparison to the control in BT and PW intestines. This implies that bacterial balance in shrimp intestines was disrupted in the presence of V. harveyi. The barcoded pyrosequencing analysis showed the similar bacterial community structures in intestines of BT and PW shrimp under a normal condition. However, during the time course exposure to V. harveyi, the relative abundance of bacteria belong to Vibrio genus was higher in the BT intestines at 12h after the exposure, whereas relative abundance of vibrios was more stable in PW intestines. The principle coordinates analysis based on weighted-UniFrac analysis showed that intestinal bacterial population in the BT shrimp lost their ability to restore their bacterial balance during the 72-h period of exposure to the pathogen, while the PW shrimp were able to reestablish their bacterial population to resemble those seen in the unexposed control group. This observation of bacterial disruption might correlate to different mortality rates observed between the two shrimp species

  19. Unusual adaptive, cross protection responses and growth phase resistance against peroxide killing in a bacterial shrimp pathogen, Vibrio harveyi.

    Science.gov (United States)

    Vattanaviboon, P; Mongkolsuk, S

    2001-06-12

    Oxidant induced protection against peroxide killing was investigated in a prawn bacterial pathogen, Vibrio harveyi. Exposure to 250 microM H(2)O(2) induced adaptive protection against subsequent exposure to killing concentrations of H(2)O(2). In addition, 200 microM t-butyl hydroperoxide (tBOOH) induced cross protection to H(2)O(2) killing. On the other hand, peroxide pretreatment did not induce protection against tBOOH killing. Peroxide induced adaptive and cross protection responses required new protein synthesis and were abolished by addition of a protein synthesis inhibitor. Pretreatments of V. harveyi with 250 microM H(2)O(2) and 200 microM tBOOH induced an increase in peroxide scavenging enzymes, catalase and alkyl hydroperoxide reductase subunit C. In addition, stationary phase cells of V. harveyi were more resistant to H(2)O(2) and iodoacetamide killing but highly susceptible to tBOOH killing compared to exponential phase cells. Many aspects of the oxidative stress response of V. harveyi are different from those of other bacteria and these factors may be important for bacterial survival in the environment and during interactions with host shrimp.

  20. Luminous vibriosis in rock lobster Jasus verreauxi (Decapoda: Palinuridae) phyllosoma larvae associated with infection by Vibrio harveyi.

    Science.gov (United States)

    Diggles, B K; Moss, G A; Carson, J; Anderson, C D

    2000-11-14

    Studies were conducted to determine the cause of outbreaks of luminous vibriosis in phyllosoma larvae of the packhorse rock lobster Jasus verreauxi reared in an experimental culture facility. On 2 separate occasions mortalities of up to 75% over a period of 4 wk were observed in 4th to 5th and 8th to 10th instar phyllosomas at water temperatures of 20 and 23 degrees C, respectively. Affected larvae became opaque, exhibited small red spots throughout the body and pereiopods, and were faintly luminous when viewed in the dark. Histopathology showed that the gut and hepatopancreas tubules of moribund phyllosomas contained massive bacterial plaques. The hepatopancreas tubules of moribund larvae were atrophic and some contained necrotic cells sloughed into the lumen. Dense, pure cultures of a bacterium identified as Vibrio harveyi were isolated from moribund larvae. The disease syndrome was reproduced by in vivo challenge and V. harveyi was successfully reisolated from diseased larvae after apparently healthy larvae were exposed by immersion to baths of more than 10(4) V. harveyi ml(-1) at 24 degrees C. Injured larvae were more susceptible to infection than were healthy larvae. Survival of larvae experimentally and naturally exposed to V. harveyi was improved when antibiotics were administered via bath exposures.

  1. Effect of the synthetic cannabinoid HU-210 on quorum sensing and on the production of quorum sensing-mediated virulence factors by Vibrio harveyi

    OpenAIRE

    Soni, Divya; Smoum, Reem; Breuer, Aviva; Mechoulam, Raphael; Steinberg, Doron

    2015-01-01

    Background Bacterial populations communicate through the cell density-dependent mechanism of quorum sensing (QS). Vibrio harveyi, one of the best studied model organisms for QS, was used to explore effects of the synthetic cannabinoid HU-210 on QS and different QS-regulated physiological processes in bacteria. Results Analysis of QS-regulated bioluminescence in wild-type and mutant strains of V. harveyi revealed that HU-210 affects the autoinducer-2 (AI-2) pathway, one of three known QS casca...

  2. Intercellular signalling in Vibrio harveyi: sequence and function of genes regulating expression of luminescence.

    Science.gov (United States)

    Bassler, B L; Wright, M; Showalter, R E; Silverman, M R

    1993-08-01

    Density-dependent expression of luminescence in Vibrio harveyi is regulated by the concentration of an extracellular signal molecule (autoinducer) in the culture medium. A recombinant clone that restored function to one class of spontaneous dim mutants was found to encode functions necessary for the synthesis of, and response to, a signal molecule. Sequence analysis of the region encoding these functions revealed three open reading frames, two (luxL and luxM) that are required for production of an autoinducer substance and a third (luxN) that is required for response to this signal substance. The LuxL and LuxM proteins are not similar in amino acid sequence to other proteins in the database, but the LuxN protein contains regions of sequence resembling both the histidine protein kinase and the response regulator domains of the family of two-component, signal transduction proteins. The phenotypes of mutants with luxL, luxM and luxN defects indicated that an additional signal-response system controlling density-dependent expression of luminescence remains to be identified.

  3. Illuminating cell signaling: Using Vibrio harveyi in an introductory biology laboratory.

    Science.gov (United States)

    Hrizo, Stacy L; Kaufmann, Nancy

    2009-05-01

    Cell signaling is an essential cellular process that is performed by all living organisms. Bacteria communicate with each other using a chemical language in a signaling pathway that allows bacteria to evaluate the size of their population, determine when they have reached a critical mass (quorum sensing), and then change their behavior in unison to carry out processes that require many cells acting together to be effective. Here, we describe a laboratory exercise in which the students observe the induction of bioluminescence or light production as an output of the quorum sensing pathway in Vibrio harveyi. Using both wildtype and mutant bacterial strains they explore the induction of community behavior via cell-cell communication by determining whether there is a correlation between the density of the bacterial population and the production of light by the bacterial culture. Using data from a cross-feeding assay the students make predictions about the identity of their strains and directly test these predictions using conditioned media from various liquid cultures. This two part exercise is designed for an introductory biology course to begin familiarizing students with collecting data, making predictions based upon the data and directly testing their hypotheses using a model organism with a cell signaling pathway that has a simple visual output: light production. Copyright © 2009 International Union of Biochemistry and Molecular Biology, Inc.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2000-01-05

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

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

    Science.gov (United States)

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

    2000-01-05

    One possible alternative to current fuel hydrodesulfurization methods is the use of microorganisms to remove sulfur compounds. Biodesulfurization requires much milder processing conditions, gives higher specificity, and does not require molecular hydrogen. In the present work we have produced two compatible plasmids: pDSR3, which allows Escherichia coli to convert dibenzothiophene (DBT) to hydroxybiphenyl (HBP), and pDSR2, which produces a Vibrio harveyi flavin oxidoreductase. We show that the flavin oxidoreductase enhances the rate of DBT removal when co-expressed in vivo with the desulfurization enzymes. The plasmids pDSR2 and pDSR3 were co-expressed in growing cultures. The expression of oxidoreductase caused an increase in the rate of DBT removal but a decrease in the rate of HBP production. The maximum rate of DBT removal was 8 mg/h. g dry cell weight. Experiments were also conducted using resting cells with the addition of various carbon sources. It was found that the addition of glucose or glycerol to cultures with oxidoreductase expression produced the highest DBT removal rate (51 mg/h. g dry cell weight). The culture with acetate and no oxidoreductase expression had the highest level of HBP production. For all carbon sources, the DBT removal rate was faster and the HBP generation rate slower with the expression of the oxidoreductase. Analysis of desulfurization intermediates indicates that the last enzyme in the pathway may be limiting. Copyright 2000 John Wiley & Sons, Inc.

  6. Construction of a stable GFP-tagged Vibrio harveyi strain for bacterial dynamics analysis of abalone infection.

    Science.gov (United States)

    Travers, Marie-Agnès; Barbou, Annaïck; Le Goïc, Nelly; Huchette, Sylvain; Paillard, Christine; Koken, Marcel

    2008-12-01

    Vibrio harveyi is a bacterial marine pathogen that can cause fatal disease in a large range of vertebrates and invertebrates, including the commercially important marine gastropod, Haliotis tuberculata. Since 1997, strains of this bacterium have regularly been causing high mortalities in farmed and wild abalone populations. The way in which the pathogen enters into abalone and the disease transmission mechanisms are thus far unknown. Therefore, a pathogenic strain, ORM4, was green fluorescent protein-tagged and validated both for its growth characteristics and for its virulence as a genuine model for abalone disease. The strain allows V. harveyi quantification by flow cytometry in seawater and in abalone haemolymph as well as the in situ detection of the parasite inside abalone tissues.

  7. Activity, Abundance, and Localization of Quorum Sensing Receptors in Vibrio harveyi.

    Science.gov (United States)

    Lorenz, Nicola; Shin, Jae Yen; Jung, Kirsten

    2017-01-01

    Quorum sensing (QS) is a process enabling a bacterial population to communicate via small molecules called autoinducers (AIs). This intercellular communication process allows single cells to synchronize their behavior within a population. The marine bacterium Vibrio harveyi ATCC BAA-1116 channels the information of three AI signals into one QS cascade. Three receptors perceive these AIs, the hybrid histidine kinases LuxN, Lux(P)Q and CqsS, to transduce the information to the histidine phosphotransfer (HPt) protein LuxU via phosphorelay, and finally to the response regulator LuxO. Hence, the level of phosphorylated LuxO depends on the AI concentrations. The phosphorylated LuxO (P-LuxO) controls the expression of small regulatory RNAs (sRNAs), which together with the RNA chaperon Hfq, destabilize the transcript of the master regulator luxR . LuxR is responsible for the induction and repression of several genes (e.g., for bioluminescence, exoprotease and siderophore production). In vivo studies with various mutants have demonstrated that the ratio between kinase and phosphatase activities of the individual QS receptors and therefore the P-LuxO/LuxO ratio is crucial not only for the output strength but also for the degree of noise. This study was undertaken to better understand the inherent design principles of this complex signaling cascade, which allows sensing and integration of different signals, but also the differentiated output in individual cells. Therefore, we quantitatively analyzed not only the enzymatic activities, but also the abundance and localization of the three QS receptors. We found that LuxN presents the highest capacity to phosphorylate LuxU, while the phosphatase activity was comparable to LuxQ and CqsS in vitro . In whole cells the copy number of LuxN was higher than that of LuxQ and CqsS, and further increased in the late exponential growth phase. Microscopy experiments indicate that LuxN and LuxQ form independent clusters. Altogether, these

  8. VanT, a homologue of Vibrio harveyi LuxR, regulates serine, metalloprotease, pigment, and biofilm production in Vibrio anguillarum.

    Science.gov (United States)

    Croxatto, Antony; Chalker, Victoria J; Lauritz, Johan; Jass, Jana; Hardman, Andrea; Williams, Paul; Cámara, Miguel; Milton, Debra L

    2002-03-01

    Vibrio anguillarum possesses at least two N-acylhomoserine lactone (AHL) quorum-sensing circuits, one of which is related to the luxMN system of Vibrio harveyi. In this study, we have cloned an additional gene of this circuit, vanT, encoding a V. harveyi LuxR-like transcriptional regulator. A V. anguillarum Delta vanT null mutation resulted in a significant decrease in total protease activity due to loss of expression of the metalloprotease EmpA, but no changes in either AHL production or virulence. Additional genes positively regulated by VanT were identified from a plasmid-based gene library fused to a promoterless lacZ. Three lacZ fusions (serA::lacZ, hpdA-hgdA::lacZ, and sat-vps73::lacZ) were identified which exhibited decreased expression in the Delta vanT strain. SerA is similar to 3-phosphoglycerate dehydrogenases and catalyzes the first step in the serine-glycine biosynthesis pathway. HgdA has identity with homogentisate dioxygenases, and HpdA is homologous to 4-hydroxyphenylpyruvate dioxygenases (HPPDs) involved in pigment production. V. anguillarum strains require an active VanT to produce high levels of an L-tyrosine-induced brown color via HPPD, suggesting that VanT controls pigment production. Vps73 and Sat are related to Vibrio cholerae proteins encoded within a DNA locus required for biofilm formation. A V. anguillarum Delta vanT mutant and a mutant carrying a polar mutation in the sat-vps73 DNA locus were shown to produce defective biofilms. Hence, a new member of the V. harveyi LuxR transcriptional activator family has been characterized in V. anguillarum that positively regulates serine, metalloprotease, pigment, and biofilm production.

  9. Lactic-acid bacteria increase the survival of marine shrimp, Litopenaeus vannamei, after infection with Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    Felipe do Nascimento Vieira

    2007-12-01

    Full Text Available This study evaluated the survival, post-larvae quality, and the population of bacteria in Litopenaeus vannamei after the addition of two strains of lactic-acid bacteria (2 and B6 experimentally infected by Vibrio harveyi. Fifteen hundred nauplii were distributed in 20 L capacity tanks with four replicates. The survival of control animals was lower (21% than that of animals fed with the strains B6 (50% and 2 (44%. Total bacterial population in the water and larvae, as well as of the Vibrio ssp. in water was not different among the treatments. No difference was observed in the population of Vibrio ssp. between the control larvae (5.5±0.5 log UFC/mL and that fed with strain 2 (5.4±0.1 log UFC/mL. Shrimp from control and fed with strain 2 showed significantly higher bacterial population than those fed with strain B6 (1.2±0.2 log UFC/mL. It was detected the lower load of Vibrio ssp. bacteria with potential of pathogenicity after feeding with strain B6.Moreover, these larvae showed more active behavior and low number of necrosis in relation to the control group and to that fed with strain 2.Este trabalho avaliou a adição de duas cepas de bactérias lácticas (2 e B6 na sobrevivência, qualidade de pós-larva e na população de bactérias na larvicultura de Litopenaeus vannamei experimentalmente infectado por Vibrio harveyi. Mil e quinhentos náuplios foram distribuídos em tanques de 20 L com quatro repetições. A sobrevivência dos animais controle foi menor (21% do que a dos alimentados com as cepas B6 (50% e 2 (44%. Sobrevivência de misis após desafio com V. harveyi foi maior em B6 do que nos outros tratamentos. A população total de bactérias na água e nas larvas, bem como de Vibrio ssp. na água não foi diferente entre os tratamentos. Não houve diferença, também, entre a população de Vibrio ssp. em larvas do grupo controle (5,5±0,5 log UFC/mL e larvas alimentadas com a cepa 2 (5,4±0,1 log UFC/mL. Camarões do grupo controle e

  10. MARINE SPONGE Aaptos suberitoides, IT’S POTENTIAL SOURCE OF NATURAL ANTIBACTERIAL FOR CONTROLLING Vibrio harveyi ON TIGER SHRIMP (Penaeus monodon CULTURE

    Directory of Open Access Journals (Sweden)

    Rosmiati Rosmiati

    2015-06-01

    Full Text Available The study aims to isolate and identify the natural antibacterial compounds potential from Aaptos suberitoides for Vibrio harveyi control on tiger shrimp (Penaeus monodon culture. The agar diffusion method using paper discs was used to determine the antibacterial activity of extracts (diethyl ether (DEE, butanol (BUE and aqueous (HOE and compounds successfully isolated against Vibrio harveyi. Findings showed that the antibacterial activity was concentrated in BUE with the inhibition zone of 17.2±0.1 mm. Meanwhile, two other extracts (DEE and HOE did not exhibit any antibacterial activity against V. harveyi. From the active BUE, it was successfully isolated two compounds giving a strong anti-vibrio activity with the inhibition zone of 22±0.1 mm. The IR, 1H, 13C, COSY, HMQC, HMBC, and MS spectrum analysis indicated that both active compounds identified as aaptamine (1 and 9-demethyloxyaaptamine (2. The study suggested that marine sponge A. suberitoides may have potential compounds source for controlling of V. harveyi on tiger shrimp culture.

  11. Multiple signalling systems controlling expression of luminescence in Vibrio harveyi: sequence and function of genes encoding a second sensory pathway.

    Science.gov (United States)

    Bassler, B L; Wright, M; Silverman, M R

    1994-07-01

    Density-dependent expression of luminescence in Vibrio harveyi is regulated by the concentration of extracellular signal molecules (autoinducers) in the culture medium. One signal-response system is encoded by the luxL,M,N locus. The luxL and luxM genes are required for the production of an autoinducer (probably beta-hydroxybutyl homoserine lactone), and the luxN gene is required for the response to that autoinducer. Analysis of the phenotypes of LuxL,M and N mutants indicated that an additional signal-response system also controls density sensing. We report here the identification, cloning and analysis of luxP and luxQ, which encode functions required for a second density-sensing system. Mutants with defects in luxP and luxQ are defective in response to a second autoinducer substance. LuxQ, like LuxN, is similar to members of the family of two-component, signal transduction proteins and contains both a histidine protein kinase and a response regulator domain. Analysis of signalling mutant phenotypes indicates that there are at least two separate signal-response pathways which converge to regulate expression of luminescence in V. harveyi.

  12. Expression and Quorum Sensing Regulation of Type III Secretion System Genes of Vibrio harveyi during Infection of Gnotobiotic Brine Shrimp.

    Directory of Open Access Journals (Sweden)

    H A Darshanee Ruwandeepika

    Full Text Available Type III secretion systems enable pathogens to inject their virulence factors directly into the cytoplasm of the host cells. The type III secretion system of Vibrio harveyi, a major pathogen of aquatic organisms and a model species in quorum sensing studies, is repressed by the quorum sensing master regulator LuxR. In this study, we found that during infection of gnotobiotic brine shrimp larvae, the expression levels of three type III secretion operons in V. harveyi increased within the first 12h after challenge and decreased again thereafter. The in vivo expression levels were highest in a mutant with a quorum sensing system that is locked in low cell density configuration (minimal LuxR levels and lowest in a mutant with a quorum sensing system that is locked in the high cell density configuration (maximal LuxR levels, which is consistent with repression of type III secretion by LuxR. Remarkably, in vivo expression levels of the type III secretion system genes were much (> 1000 fold higher than the in vitro expression levels, indicating that (currently unknown host factors significantly induce the type III secretion system. Given the fact that type III secretion is energy-consuming, repression by the quorum sensing master regulators might be a mechanism to save energy under conditions where it does not provide an advantage to the cells.

  13. Antibacterial Property of a Coral-Associated Bacterium Pseudoalteromonas luteoviolacea Against Shrimp Pathogenic Vibrio harveyi (In Vitro Study

    Directory of Open Access Journals (Sweden)

    OCKY KARNA RADJASA

    2005-06-01

    Full Text Available A coral-associated bacterium was successfully screened for secondary metabolites production based on PCR amplification of the nonribosomal peptide synthetase gene and was identified as closely related to Pseudoalteromonas luteoviolacea based on its 16S rDNA. The bacterium was found to inhibit the growth of shrimp pathogenic bacterium tested, Vibrio harveyi. To characterize the inhibiting metabolite, a 279 bp long DNA fragment was obtained and the deduced amino acid sequence showed conserved signature regions for peptide synthetases and revealed a high similarity to NosD (40% identity, a multifunctional peptide synthetase from Nostoc sp. GSV224, and NdaB (44% identity, a peptide synthetase module of Nodularia spumigena

  14. Antibacterial Property of a Coral-Associated Bacterium Pseudoalteromonas luteoviolacea Against Shrimp Pathogenic Vibrio harveyi (In Vitro Study

    Directory of Open Access Journals (Sweden)

    OCKY KARNA RADJASA

    2005-06-01

    Full Text Available A coral-associated bacterium was successfully screened for secondary metabolites production based on PCR amplification of the nonribosomal peptide synthetase gene and was identified as closely related to Pseudoalteromonas luteoviolacea based on its 16S rDNA.The bacterium was found to inhibit the growth of shrimp pathogenic bacterium tested, Vibrio harveyi. To characterize the inhibiting metabolite, a 279 bp long DNA fragment was obtained and the deduced amino acid sequence showed conserved signature regions for peptide synthetases and revealed a high similarity to NosD (40% identity, a multifunctional peptide synthetase from Nostoc sp. GSV224, and NdaB (44% identity, a peptide synthetase module of Nodularia spumigena.

  15. Studies on luminous, Vibrio harveyi associated with shrimp culture system rearing Penaeus monodon.

    Science.gov (United States)

    Kannapiran, E; Ravindran, J; Chandrasekar, R; Kalaiarasi, A

    2009-09-01

    Microbiological studies in a modified extensive shrimp culture system at Nambuthalai, southeast coast of India were carried out fora period of 120 days. Population dynamics and distribution profile of luminous bacteria and total heterotrophic bacteria in the water sediment and animal samples were monitored. Luminous bacteria associated with exoskeleton, gills and gut were isolated and quantified. The total heterotrophic bacterial counts ranged from 1.3 x 10(4) to 25.3 x 10(4) CFU ml(-1) in water and 1.5 x 10(6) to 26.2 x 10(6) CFU g(-1) in sediment. The V. harveyi population density varied between 0.6 x 10(4) and 8.8 x 10(4) LCFU ml(-1) in water and from 1.2 x 10(6) to 10.4 x 10(6) LCFU g(-1) sediment respectively. The gut of the animal was found to harbor high density of V. harveyi than gills and exoskeleton. The total heterotrophic bacteria and V. harveyi population density showed increasing trend during the culture period. The high V harveyi density observed in this study at the end of the culture period correlated with the outbreak of white spot disease.

  16. Selection of proteolytic bacteria with ability to inhibit Vibrio harveyi during white shrimp (Litopenaeus vannamei cultivation

    Directory of Open Access Journals (Sweden)

    Suntinanalert, P.

    2007-03-01

    Full Text Available Five isolates of bacteria with high proteolytic activity, isolated from water samples of intensive shrimp ponds in southern Thailand, were selected to test for the ability to control the shrimp pathogen Vibrioharveyi. 70 μl of each culture broth were investigated for their ability to inhibit V. harveyi using an agar well diffusion test but only one isolate W3 gave a reasonable sized inhibition zone of 21.62 mm. This zone wassimilar to that of oxolinic acid (2 μg and sulfamethoxazole (25 μg. The W3 isolate was identified as Pseudomonas sp. Shrimp cultivation in aquaria was conducted to investigate the inhibition of V. harveyi bythe isolate W3. The experiment consisted of a treatment of the shrimp culture with an inoculum of the isolate W3 and V. harveyi (biocontrol set, a positive control set (only inoculation of V. harveyi and a negativecontrol set as without inoculation. No mortality was found in the negative control. Shrimp mortality in the biocontrol set (33% was lower than that in the positive control set (40%; however, it showed no significantdifference (p>0.05. The average numbers of V. harveyi over 12 days of the biocontrol set were lower than those in the positive control set by about 1 log cycle although the numbers were not significantly different(p>0.05. The shrimp growth rate at day 32 of cultivation was in order of the biocontrol treatment (10.17% > the negative control treatment (9.44% > the positive control set (9.28%, but no significant difference (p>0.05 was observed among treatments.

  17. Using fusions with luxAB from Vibrio harveyi MAV to quantify induction and catabolite repression of the xyl operon in Staphylococcus carnosus TM300.

    Science.gov (United States)

    Sizemore, C; Geissdörfer, W; Hillen, W

    1993-03-01

    The luxA,B genes from the Gram-negative marine bacterium Vibrio harveyi MAV were used in Staphylococcus carnosus TM300 as a reporter system for regulated expression of xylose utilization. The luciferase genes were fused to the xyl operon from Staphylococcus xylosus C2a. Expression of bioluminescence was induced through addition of xylose and repressed in the presence of glucose. A method to quantitate bioluminescence directly from the culture is described.

  18. Laboratory simulation reveals significant impacts of ocean acidification on microbial community composition and host-pathogen interactions between the blood clam and Vibrio harveyi.

    Science.gov (United States)

    Zha, Shanjie; Liu, Saixi; Su, Wenhao; Shi, Wei; Xiao, Guoqiang; Yan, Maocang; Liu, Guangxu

    2017-12-01

    It has been suggested that climate change may promote the outbreaks of diseases in the sea through altering the host susceptibility, the pathogen virulence, and the host-pathogen interaction. However, the impacts of ocean acidification (OA) on the pathogen components of bacterial community and the host-pathogen interaction of marine bivalves are still poorly understood. Therefore, 16S rRNA high-throughput sequencing and host-pathogen interaction analysis between blood clam (Tegillarca granosa) and Vibrio harveyi were conducted in the present study to gain a better understanding of the ecological impacts of ocean acidification. The results obtained revealed a significant impact of ocean acidification on the composition of microbial community at laboratory scale. Notably, the abundance of Vibrio, a major group of pathogens to many marine organisms, was significantly increased under ocean acidification condition. In addition, the survival rate and haemolytic activity of V. harveyi were significantly higher in the presence of haemolymph of OA treated T. granosa, indicating a compromised immunity of the clam and enhanced virulence of V. harveyi under future ocean acidification scenarios. Conclusively, the results obtained in this study suggest that future ocean acidification may increase the risk of Vibrio pathogen infection for marine bivalve species, such as blood clams. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Quorum sensing influences Vibrio harveyi growth rates in a manner not fully accounted for by the marker effect of bioluminescence.

    Science.gov (United States)

    Nackerdien, Zeena E; Keynan, Alexander; Bassler, Bonnie L; Lederberg, Joshua; Thaler, David S

    2008-02-27

    The light-emitting Vibrios provide excellent material for studying the interaction of cellular communication with growth rate because bioluminescence is a convenient marker for quorum sensing. However, the use of bioluminescence as a marker is complicated because bioluminescence itself may affect growth rate, e.g. by diverting energy. The marker effect was explored via growth rate studies in isogenic Vibrio harveyi (Vh) strains altered in quorum sensing on the one hand, and bioluminescence on the other. By hypothesis, growth rate is energy limited: mutants deficient in quorum sensing grow faster because wild type quorum sensing unleashes bioluminescence and bioluminescence diverts energy. Findings reported here confirm a role for bioluminescence in limiting Vh growth rate, at least under the conditions tested. However, the results argue that the bioluminescence is insufficient to explain the relationship of growth rate and quorum sensing in Vh. A Vh mutant null for all genes encoding the bioluminescence pathway grew faster than wild type but not as fast as null mutants in quorum sensing. Vh quorum sensing mutants showed altered growth rates that do not always rank with their relative increase or decrease in bioluminescence. In addition, the cell-free culture fluids of a rapidly growing Vibrio parahaemolyticus (Vp) strain increased the growth rate of wild type Vh without significantly altering Vh's bioluminescence. The same cell-free culture fluid increased the bioluminescence of Vh quorum mutants. The effect of quorum sensing on Vh growth rate can be either positive or negative and includes both bioluminescence-dependent and independent components. Bioluminescence tends to slow growth rate but not enough to account for the effects of quorum sensing on growth rate.

  20. Quorum sensing influences Vibrio harveyi growth rates in a manner not fully accounted for by the marker effect of bioluminescence.

    Directory of Open Access Journals (Sweden)

    Zeena E Nackerdien

    2008-02-01

    Full Text Available The light-emitting Vibrios provide excellent material for studying the interaction of cellular communication with growth rate because bioluminescence is a convenient marker for quorum sensing. However, the use of bioluminescence as a marker is complicated because bioluminescence itself may affect growth rate, e.g. by diverting energy.The marker effect was explored via growth rate studies in isogenic Vibrio harveyi (Vh strains altered in quorum sensing on the one hand, and bioluminescence on the other. By hypothesis, growth rate is energy limited: mutants deficient in quorum sensing grow faster because wild type quorum sensing unleashes bioluminescence and bioluminescence diverts energy. Findings reported here confirm a role for bioluminescence in limiting Vh growth rate, at least under the conditions tested. However, the results argue that the bioluminescence is insufficient to explain the relationship of growth rate and quorum sensing in Vh. A Vh mutant null for all genes encoding the bioluminescence pathway grew faster than wild type but not as fast as null mutants in quorum sensing. Vh quorum sensing mutants showed altered growth rates that do not always rank with their relative increase or decrease in bioluminescence. In addition, the cell-free culture fluids of a rapidly growing Vibrio parahaemolyticus (Vp strain increased the growth rate of wild type Vh without significantly altering Vh's bioluminescence. The same cell-free culture fluid increased the bioluminescence of Vh quorum mutants.The effect of quorum sensing on Vh growth rate can be either positive or negative and includes both bioluminescence-dependent and independent components. Bioluminescence tends to slow growth rate but not enough to account for the effects of quorum sensing on growth rate.

  1. Assimilable organic carbon (AOC) in soil water extracts using Vibrio harveyi BB721 and its implication for microbial biomass.

    Science.gov (United States)

    Ma, Jincai; Ibekwe, A Mark; Wang, Haizhen; Xu, Jianming; Leddy, Menu; Yang, Ching-Hong; Crowley, David E

    2012-01-01

    Assimilable organic carbon (AOC) is commonly used to measure the growth potential of microorganisms in water, but has not yet been investigated for measuring microbial growth potential in soils. In this study, a simple, rapid, and non-growth based assay to determine AOC in soil was developed using a naturally occurring luminous strain Vibrio harveyi BB721 to determine the fraction of low molecular weight organic carbon in soil water extract. Calibration of the assay was achieved by measuring the luminescence intensity of starved V. harveyi BB721 cells in the late exponential phase with a concentration range from 0 to 800 µg l(-1) glucose (equivalent to 0-16.0 mg glucose C kg(-1) soil) with the detection limit of 10 µg l(-1) equivalent to 0.20 mg glucose C kg(-1) soil. Results showed that bioluminescence was proportional to the concentration of glucose added to soil. The luminescence intensity of the cells was highly pH dependent and the optimal pH was about 7.0. The average AOC concentration in 32 soils tested was 2.9±2.2 mg glucose C kg(-1). Our data showed that AOC levels in soil water extracts were significantly correlated (Pgrowth bioluminescence based assay. Understanding the levels of AOC in soil water extract provides new insights into our ability to estimate the most available carbon pool to bacteria in soil that may be easily assimilated into cells for many metabolic processes and suggest possible the links between AOC, microbial regrowth potential, and microbial biomass in soils.

  2. Assimilable organic carbon (AOC in soil water extracts using Vibrio harveyi BB721 and its implication for microbial biomass.

    Directory of Open Access Journals (Sweden)

    Jincai Ma

    Full Text Available Assimilable organic carbon (AOC is commonly used to measure the growth potential of microorganisms in water, but has not yet been investigated for measuring microbial growth potential in soils. In this study, a simple, rapid, and non-growth based assay to determine AOC in soil was developed using a naturally occurring luminous strain Vibrio harveyi BB721 to determine the fraction of low molecular weight organic carbon in soil water extract. Calibration of the assay was achieved by measuring the luminescence intensity of starved V. harveyi BB721 cells in the late exponential phase with a concentration range from 0 to 800 µg l(-1 glucose (equivalent to 0-16.0 mg glucose C kg(-1 soil with the detection limit of 10 µg l(-1 equivalent to 0.20 mg glucose C kg(-1 soil. Results showed that bioluminescence was proportional to the concentration of glucose added to soil. The luminescence intensity of the cells was highly pH dependent and the optimal pH was about 7.0. The average AOC concentration in 32 soils tested was 2.9±2.2 mg glucose C kg(-1. Our data showed that AOC levels in soil water extracts were significantly correlated (P<0.05 with microbial biomass determined as microbial biomass carbon, indicating that the AOC concentrations determined by the method developed might be a good indicator of soil microbial biomass. Our findings provide a new approach that may be used to determine AOC in environmental samples using a non-growth bioluminescence based assay. Understanding the levels of AOC in soil water extract provides new insights into our ability to estimate the most available carbon pool to bacteria in soil that may be easily assimilated into cells for many metabolic processes and suggest possible the links between AOC, microbial regrowth potential, and microbial biomass in soils.

  3. Effect of Lactobacillus plantarum isolated from digestive tract of wild shrimp on growth and survival of white shrimp (Litopenaeus vannamei) challenged with Vibrio harveyi.

    Science.gov (United States)

    Kongnum, Khanitta; Hongpattarakere, Tipparat

    2012-01-01

    Two hundred and two strains of lactic acid bacteria (LAB) isolated from digestive tracts of cultivated and wild adult shrimp, including Litopenaeus vannamei, Metapenaeus brevicornis and Penaeus merguiensis were selected based on their antibacterial activity against Vibrio harveyi. LAB strain of MRO3.12 exhibiting highest reduction of V. harveyi was identified as Lactobacillus plantarum MRO3.12 based on the nucleotide sequence of its 16S rDNA, which showed 99% (780/786 bp) homology to L. plantarum strain L5 (GenBank accession number DQ 239698.1). Co-cultivation of V. harveyi and L. plantarum MRO3.12 showed complete reduction of V. harveyi at 24 h under aerobic and anaerobic conditions, whereas L. plantarum increased from 5.29 to 9.47 log CFU ml(-1). After 6-week feeding trial with L. plantarum supplemented diet, white shrimp (L. vannamei) exhibited significant differences (p < 0.05) in relative growth rate (% RGR), feed conversion ratio (FCR) and survival compared to the control group fed with non-supplemented diet. LAB-fed group showed 98.89% survival, whereas only 68.89% survival was observed in the control group. LAB from the digestive tract of probiotic-fed shrimp showed higher level of 5.0 ± 0.14 log CFU/g than the non-supplemented ones (3.34 ± 0.21 log CFU/g). However, total bacterial and non-fermenting vibrios counts decreased in shrimps fed on L. plantarum. Ten days after infection with V. harveyi (5.3-5.5 log CFU ml(-1)), significant survival (p < 0.05) of 77% was observed in LAB supplemented shrimp, while only 67% survival was observed in the control. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. Induction of peroxide and superoxide protective enzymes and physiological cross-protection against peroxide killing by a superoxide generator in Vibrio harveyi.

    Science.gov (United States)

    Vattanaviboon, Paiboon; Panmanee, Warunya; Mongkolsuk, Skorn

    2003-04-11

    Vibrio harveyi is a causative agent of destructive luminous vibriosis in farmed black tiger prawn (Penaeus monodon). V. harveyi peroxide and superoxide stress responses toward elevated levels of a superoxide generated by menadione were investigated. Exposure of V. harveyi to sub-lethal concentrations of menadione induced high expression of genes in both the OxyR regulon (e.g., a monofunctional catalase or KatA and an alkyl hydroperoxide reductase subunit C or AhpC), and the SoxRS regulon (e.g., a superoxide dismutase (SOD) and a glucose-6-phosphate dehydrogenase). V. harveyi expressed two detectable, differentially regulated SOD isozymes, [Mn]-SOD and [Fe]-SOD. [Fe]-SOD was expressed constitutively throughout the growth phase while [Mn]-SOD was expressed at the stationary phase and could be induced by a superoxide generator. Physiologically, pre-treatment of V. harveyi with menadione induced cross-protection against subsequent exposure to killing concentrations of H(2)O(2). This induced cross-protection required newly synthesized proteins. However, the treatment did not induce significant protection against exposures to killing concentrations of menadione itself or cross-protect against an organic hydroperoxide (tert-butyl hydroperoxide). Unexpectedly, growing V. harveyi in high-salinity media induced protection against menadione killing. This protection was independent of SOD induction. Stationary-phase cells were more resistant to menadione killing than exponential-phase cells. The induction of oxidative stress protective enzymes and stress-altered physiological responses could play a role in the survival of this bacterium in the host marine crustaceans.

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

    Science.gov (United States)

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

    2005-09-01

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

  6. Establishment and Preliminary Application of the Duplex PCR to Detection of Vibrio vulnificus and Vibrio harveyi%创伤弧菌和哈氏弧菌双重PCR检测方法的建立及初步应用

    Institute of Scientific and Technical Information of China (English)

    陈福艳; 王瑞; 李丽萍; 韦明利; 程光平; 欧阳贤华; 陈晓汉; 梁万文; 韦友传; 黄婷; 杨学明; 陈明

    2013-01-01

    As Vibrio vulnificus and Vibrio harveyi mainly affects marine cultured animals, it is necessary to establish a rapid detection method. According to the cytolysin gene sequence of V. vulnificus and toxR gene sequence of V. harveyi, two pairs of primers were designed and synthesized. The diagnosis of duplex polymerase chain reaction (PCR) for rapid identification of V. vulnificus and V. harveyi was established by optimization of PCR conditions and by testing specificity and sensitiveness. The duplex PCR test results showed that two specific and amplified fragments were produced which sizes were 128 bp for V. vulnificus and 211 bp for V. harveyi, and no cross-reaction tests were done with Streptococcus agalactiae, Streptococcus iniae, Bibrio parahemolyticus, and Vibrio damsela. The lowest concentrations of 12 cfu/mL for V. vulnificus and 18 cfu /mL for V. harveyi could be detected. The duplex PCR would be a specific and sensitive method which was timesaving, laborsaving, quick and high-efficient for identification of duplex V. vulnificus and V. harveyi.%  根据创伤弧菌(Vibrio vulnificus)的溶细胞毒素基因序列和哈氏弧菌(Vibrio harveyi)的toxR基因序列,分别设计并合成两对特异性引物,通过PCR反应条件优化,测试两种菌的特异性和敏感性,建立双重PCR方法,同时快速检测V. vulnificus和V. harveyi。结果表明:纯培养V. vulnificus和V. harveyi的检测灵敏度分别是12 cfu/mL和18 cfu/mL,与无乳链球菌、海豚链球菌、副溶血弧菌及美人发光杆菌无交叉反应;此PCR 检测方法具有良好的特异性、敏感性,具快速、高效等优点,对细菌V. vulnificus和V. harveyi诊断与防治具有较好的临床应用性。

  7. Effect of the synthetic cannabinoid HU-210 on quorum sensing and on the production of quorum sensing-mediated virulence factors by Vibrio harveyi.

    Science.gov (United States)

    Soni, Divya; Smoum, Reem; Breuer, Aviva; Mechoulam, Raphael; Steinberg, Doron

    2015-08-12

    Bacterial populations communicate through the cell density-dependent mechanism of quorum sensing (QS). Vibrio harveyi, one of the best studied model organisms for QS, was used to explore effects of the synthetic cannabinoid HU-210 on QS and different QS-regulated physiological processes in bacteria. Analysis of QS-regulated bioluminescence in wild-type and mutant strains of V. harveyi revealed that HU-210 affects the autoinducer-2 (AI-2) pathway, one of three known QS cascades of V. harveyi. Furthermore, QS-mediated biofilm formation and swimming motility in the mutant strain BB152 (AI-1(-), AI-2(+)) were significantly reduced in the presence of HU-210. HU-210 inhibited QS-mediated virulence factor production without any inhibitory effect on bacterial growth. It also alters the expression of several genes, which are regulated by QS, specifically downregulating the genes of the AI-2 QS cascade. First evidence is being provided for interference of bacterial signal-transduction systems by a synthetic cannabinoid. The effect of HU-210 was specific to the AI-2 cascade in V. harveyi. AI-2 is known as a "universal autoinducer" and interference with its activity opens a broad spectrum of applications for synthetic cannabinoids in future research as a potential anti-QS agent.

  8. Identification of Vibrio harveyi as a causative bacterium for a tail rot disease of sea bream Sparus aurata from research hatchery in Malta.

    Science.gov (United States)

    Haldar, S; Maharajan, A; Chatterjee, S; Hunter, S A; Chowdhury, N; Hinenoya, A; Asakura, M; Yamasaki, S

    2010-10-20

    A bacterial disease was reported from gilthead sea bream (Sparus aurata) within a hatchery environment in Malta. Symptoms included complete erosion of tail, infection in the eye, mucous secretion and frequent mortality. A total of 540 strains were initially isolated in marine agar from different infected body parts and culture water sources. Subsequently 100 isolates were randomly selected, identified biochemically and all were found to be Vibrio harveyi-related organisms; finally from 100 isolates a total of 13 numbers were randomly selected and accurately identified as V. harveyi by 16S rRNA gene sequencing and species-specific PCR. Ribotyping of these strains with HindIII revealed total of six clusters. In vivo challenge study with representative isolates from each cluster proved two clusters each were highly pathogenic, moderately pathogenic and non-pathogenic. All 13 isolates were positive for hemolysin gene, a potential virulence factor. Further analysis revealed probably a single copy of this gene was encoded in all isolates, although not in the same locus in the genome. Although V. harveyi was reported to be an important pathogen for many aquatic organisms, to our knowledge this might be the first report of disease caused by V. harveyi and their systematic study in the sea bream hatchery from Malta. Copyright © 2009 Elsevier GmbH. All rights reserved.

  9. Molecular genetic diversity of Vibrio harveyi%哈维氏弧菌的分子遗传多样性研究

    Institute of Scientific and Technical Information of China (English)

    徐芝亮; 吴灶和; 简纪常; 黄郁葱

    2012-01-01

    Vibrio harveyi is known to be a major pathogen in mariculture animal systems. Disease outbreaks attributed to V. harveyi have been reported in main mariculture animal farm production. In recent years,mass mortalities of many cultured kinds of maricultured fishes, such as Litopenaeus vannamei, Pseudosciaena crocea, and Lutjanus sanguineus have occurred frequently infected by V. harveyi. In this article the 101 isolates of V. harveyi were subjected to random amplified polymorphic DNA (RAPD)-PCR and BOX-PCR analysis to investigate the genetic variability among V. harveyi strains. A total of 10 RAPD primers were designed for their specificity in detecting V. harveyi, and only the primer PM2 was highly reproducible and found suitable to use in RAPD-PCR. The genetic diversity among V. harveyi isolates assessed by RAPD-PCR by PM2 primer yielded 15 different RAPD patterns which clustered the isolates into 18 groups at 65% similarity level. Similarly, BOX-PCR clustered the 20 patterns into 15 groups at 40% similarity. However, with RAPD-PCR and BOX-PCR, 101 V. harveyi could be divided into 18 groups at 50% similarity. We could obtain the accurate results of molecular genetic diversity by both RAPD-PCR and BOX-PCR.%运用RAPD分型和BOX分型分子方法研究患病的海水鱼类体内和海水环境的101株哈维氏弧菌的分子遗传多样性.结果显示,从10个RAPD引物中发现只有PM2引物的分辨率和重复性较好,出现15条不同RAPD条带,在相似度65%下,101株哈维氏弧菌可以分为18种不同型;BOX分型出现20条不同的条带,在相似度为40%的情况下,可以把101株哈维式弧菌分15种不同的型;综合RAPD PM2和BOX分型,发现在相似度50%下,101株哈维氏弧菌可分18种型.综合RAPD和BOX分型数据可以很好地克服各自的缺点,得到更加准确的分型结果.

  10. Interaction of Haematopoietic Tissue Cultures of the Dublin Bay Prawn,Nephrops Norvegicus(L.),with the Causal Agent of Luminous Vibriosis Vibrio Harveyi

    Institute of Scientific and Technical Information of China (English)

    Mulford A.L.; ZHANG X.H.; XU H. S.; Austin B.

    2002-01-01

    Vibrio harveyi cells (dose = > 103 cells mL 1) and extracellular products (ECP; >25μg m L-1 of total protein concentration) destroyed haematopoietic cultures of Nephrops norvegicus within 24 h of exposure. Cytopathic effects (CPE)started after 4 h of exposure to the bacterial cells, with some granularity in the cytoplasm, mostly in cells in the outer periphery of the explant growth. At the end of the infection, a considerable number of nuclei remained attached to the substrate,apparently unaffected. Following exposure to ECP, initial deterioration was observed at 2 h with the presence of granularity in the cytoplasm of< 20% cells, and few cells displayed small vacuoles around the nuclei. Parallel results were obtained using whole animal experiments, with V. harveyi cells being lethal to nephrops within 24 h.

  11. Interaction of haematopoietic tissue cultures of the Dublin Bay Prawn, Nephrops norvegicus (L.), with the causal agent of luminous vibriosis Vibrio harveyi

    Science.gov (United States)

    Mulford, A. L.; Zhang, X. H.; Xu, H. S.; Austin, B.

    2002-04-01

    Vibrio harveyi cells (dose—25 μmg mL-1 of total protein concentration) destroyed haematopoietic cultures of Nephrops norvegicus within 24 h of exposure. Cytopathic effects (CPE) started after 4h of exposure to the bacterial cells, with some granularity in the cytoplasm, mostly in cells in the outer periphery of the explant growth. At the end of the infection, a considerable number of nuclei remained attached to the substrate, apparently unaffected. Following exposure to ECP, initial deterioration was observed at 2 h with the presence of granularity in the cytoplasm of<20% cells, and few cells displayed small vacuoles around the nuclei. Parallel results were obtained using whole animal experiments, with V. harveyi cells being lethal to nephrops within 24 h.

  12. Effects of Microencapsulated Synbiotic Administration at Different Dosages against heavy co-infection of White Spot Disease (WSD and Vibrio harveyi in Pacific White Shrimp (Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Yunarty Yunarty

    2016-12-01

    Full Text Available White spot disease (WSD is one of infectious disease in shrimp caused by white spot syndrome virus (WSSV. This study aimed to determine the dosage immunological effects and growth performances of microencapsulated synbiotic (Bacillus NP5 and mannan oligosaccharide at different dosages on Pacific white shrimp.  The microencapsulated synbiotic   was administered as feed supplementation  against the co-infection of   WSSV and Vibrio harveyi. Synbiotic was encapsulated by spray drying method, further feed supplemented to Pacific white shrimp for 30 days at a  dosages of 0.5% (A, 1% (B, 2% (C and control treatments, i.e. without any microencapsulated synbiotic administration as positive control (D and negative control (E. The challenge test was performed on day 30 after feeding supplementation, then the experimental shrimps were injected by WSSV intramuscularly   at the infective dosage of 104 copies.-ml-1. Afterwards,   24 hours after WSSV injection the shrimps were immersed in water contained cells suspension of V. harveyi  at the cells population dosage of 106 CFU-.ml-1. All synbiotic treatments showed better results with the values of Total Haemocyte Count (THC, Phenoloxidase (PO and Respiratory Burst (RB, were higher (P<0.05 compared to positive control. The specific growth rates (SGR of A, B and C showed higher than both controls of D and E. The feed conversion ratio (FCR value of synbiotic treatments were lower (P<0.05 than both controls. However, the administration of microencapsulated synbiotic have not been able to prevent heavy impact of WSSV and V. harveyi co-infection due to lower SR and mortality pattern which continued to increase.   Keywords: Synbiotic, Litopenaeus vannamei, WSSV, Vibrio harveyi, co-infection

  13. Effect of probiotic supplemented diet on marine shrimp survival after challenge with Vibrio harveyi Efeito do uso de dieta suplementada com o probiótico sobre a sobrevivência de camarões marinhos após o desafio com Vibrio harveyi

    OpenAIRE

    F.N. Vieira; C.C. Buglione; J.P.L. Mouriño; A. Jatobá; M.L. Martins; D.D. Schleder; E.R. Andreatta; M.A. Barraco; L.A. Vinatea

    2010-01-01

    The effect of a Lactobacillus plantarum-supplemented diet on shrimp growth, digestive tract bacterial microbiota, survival, and some hemato-immunological parameters after an experimental challenge with Vibrio harveyi was studied. No difference (P>0.05) was observed in shrimp survival (80±6%) and final weight (6.63±0.56g) after 60 days feeding trial. Total bacteria count and Vibrio spp. count in the digestive tract were not diferent (P>0.05) until day 40, but they were lower (P0.05) than the c...

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

    Science.gov (United States)

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

    2007-07-01

    To evaluate the effect of Vibrio harveyi strains on the growth rate of the gnotobiotically cultured rotifer Brachionus plicatilis, and to establish whether quorum sensing is involved in the observed phenomena. Gnotobiotic B. plicatilis sensu strictu, obtained by hatching glutaraldehyde-treated amictic eggs, were used as test organisms. Challenge tests were performed with 11 V. harveyi strains and different quorum sensing mutants derived from the V. harveyi BB120 strain. Brominated furanone [(5Z)-4-bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone] as a quorum sensing inhibitor was tested in Brachionus challenge tests. Some V. harveyi strains, such as strain BB120, had a significantly negative effect on the Brachionus growth rate. In the challenge test with MM77, an isogenic strain of BB120 in which the two autoinducers (HAI-1 and AI-2) are both inactivated, no negative effect was observed. The effect of single mutants was the same as that observed in the BB120 strain. This indicates that both systems are responsible for the growth-retarding (GR) effect of the BB120 strain towards Brachionus. Moreover, the addition of an exogenous source of HAI-1 or AI-2 could restore the GR effect in the HAI-1 and AI-2 nonproducing mutant MM77. The addition of brominated furanone at a concentration of 2.5 mg l(-1) could neutralize the GR effect of some strains such as BB120 and VH-014. Two quorum sensing systems in V. harveyi strain BB120 (namely HAI-1 and AI-2-mediated) are necessary for its GR effect on B. plicatilis. With some other V. harveyi strains, however, growth inhibition towards Brachionus does not seem to be related to quorum sensing. Interference with the quorum sensing system might help to counteract the GR effect of some V. harveyi strains on Brachionus. However, further studies are needed to demonstrate the positive effect of halogenated furanone in nongnotobiotic Brachionus cultures and eventually, in other segments of the aquaculture industry.

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

    Science.gov (United States)

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

    2014-05-21

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

  16. Pathogenesis and virulence of Vibrio harveyi from southern part of Thailand in black tiger shrimp, Penaeus monodon Fabricius

    Directory of Open Access Journals (Sweden)

    Pecharat, J.

    2004-01-01

    Full Text Available Ten isolates of bacteria were performed from diseased black tiger shrimp collected from farms in Nakhon Si Thammarat, Patthalung, Songkhla, Satun and Patthani. All isolates belonged to Vibrio harveyi, forming rounded colonies with smooth periphery and hemolysis of red blood cells. The colonies appearedgreen and yellow on TCBS selective media and indeeded both luminescent and non-luminescent groups. To support good growth as cell proliferation, an addition of 2-8% salt to the culture media was required. Sensitivity tests against antibiotics showed that all ten isolates were sensitive to chloramphenicol, norfloxacin, trimethoprim-sulfamethoxazole, oxolinic acid, oxytetracyclin and sarafloxacin at 80, 60, 50, 40, 40 and 30%, respectively. When 12-15 g juvenile shrimp were tested for the virulence by injection, the isolates were highly virulent with OD at 640 nm in the range 0.007-0.139. The bacteria concentration of 1.60×106 - 7.27×107 CFU/ ml caused 50% juvenile mortality in ten days. The physiological changes after susceptibility included a decline in the blood cells (p<0.05, higher plasma pH (p<0.05, decrease in serum protein within 24 h (p<0.05, and a return to normal levels within 48 h. There were no differences in blood sugar levels and phenoloxidase activity between the healthy and the infected individuals. Histological study showed that at an early period of susceptibility, there were swollen tubular lumen, minor cell degeneration of the hepatic tubules and lymphoid organs and aggregation of blood cells around the degenerating cells. During 7-day susceptibility period, there was large scale cell degeneration of the hepatopancreas, lymphoid organs, gills and blood formingorgans with a great extent of blood cell aggregation and eventually mortalities occurred. During 14 day susceptibility period, there was almost entire degeneration of cells in hepatic tubules and lymphoid organs causing hepatopancreatic tubular necrosis. During this period

  17. Pleurocidin Peptide Enhances Grouper Anti-Vibrio harveyi Immunity Elicited by Poly(lactide-co-glycolide)-Encapsulated Recombinant Glyceraldehyde-3-phosphate Dehydrogenase.

    Science.gov (United States)

    Chuang, Shu-Chun; Huang, Wan-Ling; Kau, Sau-Wei; Yang, Yun-Pei; Yang, Chung-Da

    2014-05-14

    Outer membrane proteins, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), are considered immunodominant antigens for eliciting protective immunity against Vibrio harveyi, the main etiological agent of vibriosis in fish. Cationic antimicrobial peptides (AMPs), such as pleurocidin (PLE), play important roles in activating and recruiting immune cells, thereby contributing to subsequent innate and adaptive immune responses. In the present study, we aimed to use PLE peptide as a potent adjuvant to improve the immunogenicity of V. harveyi recombinant GAPDH (rGAPDH). In order to prepare a controlled-release vaccine, PLE peptide and rGAPDH protein were simultaneously encapsulated into polymeric microparticles made from the biodegradable poly(lactide-co-glycolide) (PLG) polymer. The resulting PLG-encapsulated PLE plus rGAPDH (PLG-PLE/rGAPDH) microparticles, 3.21-6.27 μm in diameter, showed 72%-83% entrapment efficiency and durably released both PLE and rGAPDH for a long 30-day period. Following peritoneal immunization in grouper (Epinephelus coioides), PLG-PLE/rGAPDH microparticles resulted in significantly higher (p PLE/rGAPDH microparticles conferred a high survival rate (85%), which was significantly higher (p PLE peptide exhibits an efficacious adjuvant effect to elicit not only improved immunity, but also enhanced protection against V. harveyi in grouper induced by rGAPDH protein encapsulated in PLG microparticles.

  18. Azadirachta indica (neem) leaf dietary effects on the immunity response and disease resistance of Asian seabass, Lates calcarifer challenged with Vibrio harveyi.

    Science.gov (United States)

    Talpur, Allah Dad; Ikhwanuddin, Mhd

    2013-01-01

    The present study was aimed to address the possible evaluation of Azadirachta indica (neem) leaf-supplemented diets on innate immune response in Asian seabass, Lates calcarifer fingerlings against Vibrio harveyi infection. Fish were fed for two weeks diets containing six graded levels of neem leaf at 0 g, 1 g, 2 g, 3 g, 4 g and 5 g per kg feed. Fish fed neem leaf-supplemented diet displayed significant differences (p growth rate (SGR) and feed conversion ratio (FCR) compared to the control group fed without neem leaf-supplemented diet. Various innate immune parameters were examined pre-challenge and post-challenge. Fish was injected intraperitoneally with a lethal dose of V. harveyi containing 10(8) cells mL(-1). Supplementation of neem leaf diet significantly increased phagocytic activity, superoxide anion production, serum lysozyme, serum bactericidal activity, serum anti-protease activity throughout the experimental period when compared with the control group. Dietary doses of neem leaf diet significantly influenced the immune parameters, haematological parameters and blood biochemical indices of treated fish. The results suggested that fish fed neem leaf-supplemented diet improved the immune system and increased survival rate in L. calcarifer fingerlings against V. harveyi infection. Copyright © 2012 Elsevier Ltd. All rights reserved.

  19. Expression of Toll-like receptors (TLR, in lymphoid organ of black tiger shrimp (Penaeus monodon in response to Vibrio harveyi infection

    Directory of Open Access Journals (Sweden)

    Mundanda Muthappa Dechamma

    2015-05-01

    Full Text Available The Toll-like receptors (TLR, being pattern recognition molecules, are a powerful first line of defense in response to pathogen invasion. They are known to play a crucial role in detecting and binding to the microbial molecule and triggering a non-specific immune response. Quantitative real time PCR (qPCR expression of the TLR gene was studied in healthy and Vibrio harveyi infected black tiger shrimp (Penaeus monodon. Lymphoid tissue expression of TLR in V. harveyi infected animals 24 h post injection showed statistically significant up regulation of the gene as compared to the control animals sham injected with phosphate buffered saline (PBS. The qPCR expression pattern of TLR at different time points in shrimp administered with the immunostimulant glucan for 6 days by oral feeding followed by challenge with V. harveyi showed statistically significant level at 48 h post bacterial challenge as compared to the control (immunostimulant treated animals sham injected with PBS. The novelty of the study is that it elicits the role of TLRs as important response proteins of the innate immune system in the shrimp.

  20. Structural and functional investigation of flavin binding center of the NqrC subunit of sodium-translocating NADH:quinone oxidoreductase from Vibrio harveyi.

    Directory of Open Access Journals (Sweden)

    Valentin Borshchevskiy

    Full Text Available Na+-translocating NADH:quinone oxidoreductase (NQR is a redox-driven sodium pump operating in the respiratory chain of various bacteria, including pathogenic species. The enzyme has a unique set of redox active prosthetic groups, which includes two covalently bound flavin mononucleotide (FMN residues attached to threonine residues in subunits NqrB and NqrC. The reason of FMN covalent bonding in the subunits has not been established yet. In the current work, binding of free FMN to the apo-form of NqrC from Vibrio harveyi was studied showing very low affinity of NqrC to FMN in the absence of its covalent bonding. To study structural aspects of flavin binding in NqrC, its holo-form was crystallized and its 3D structure was solved at 1.56 Å resolution. It was found that the isoalloxazine moiety of the FMN residue is buried in a hydrophobic cavity and that its pyrimidine ring is squeezed between hydrophobic amino acid residues while its benzene ring is extended from the protein surroundings. This structure of the flavin-binding pocket appears to provide flexibility of the benzene ring, which can help the FMN residue to take the bended conformation and thus to stabilize the one-electron reduced form of the prosthetic group. These properties may also lead to relatively weak noncovalent binding of the flavin. This fact along with periplasmic location of the FMN-binding domains in the vast majority of NqrC-like proteins may explain the necessity of the covalent bonding of this prosthetic group to prevent its loss to the external medium.

  1. A Nonluminescent and Highly Virulent Vibrio harveyi Strain Is Associated with “Bacterial White Tail Disease” of Litopenaeus vannamei Shrimp

    Science.gov (United States)

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

    2012-01-01

    Recurrent outbreaks of a disease in pond-cultured juvenile and subadult Litopenaeus vannamei shrimp in several districts in China remain an important problem in recent years. The disease was characterized by “white tail” and generally accompanied by mass mortalities. Based on data from the microscopical analyses, PCR detection and 16S rRNA sequencing, a new Vibrio harveyi strain (designated as strain HLB0905) was identified as the etiologic pathogen. The bacterial isolation and challenge tests demonstrated that the HLB0905 strain was nonluminescent but highly virulent. It could cause mass mortality in affected shrimp during a short time period with a low dose of infection. Meanwhile, the histopathological and electron microscopical analysis both showed that the HLB0905 strain could cause severe fiber cell damages and striated muscle necrosis by accumulating in the tail muscle of L. vannamei shrimp, which led the affected shrimp to exhibit white or opaque lesions in the tail. The typical sign was closely similar to that caused by infectious myonecrosis (IMN), white tail disease (WTD) or penaeid white tail disease (PWTD). To differentiate from such diseases as with a sign of “white tail” but of non-bacterial origin, the present disease was named as “bacterial white tail disease (BWTD)”. Present study revealed that, just like IMN and WTD, BWTD could also cause mass mortalities in pond-cultured shrimp. These results suggested that some bacterial strains are changing themselves from secondary to primary pathogens by enhancing their virulence in current shrimp aquaculture system. PMID:22383954

  2. Promoting resuscitation of viable but nonculturable cells of Vibrio harveyi by a resuscitation-promoting factor-like protein YeaZ.

    Science.gov (United States)

    Li, Y; Chen, J; Zhao, M; Yang, Z; Yue, L; Zhang, X

    2017-02-01

    To demonstrate the resuscitation-promoting activities of recombinant YeaZ from Vibrio harveyi SF-1. The gene of resuscitation-promoting factor YeaZ was cloned from genomic DNA of V. harveyi SF-1. The gene was expressed in Escherichia coli, and the expressed protein was purified by Ni 2+ -affinity chromatography. A yeaZ mutant was constructed by using the suicide plasmid pNQ705 with homologous recombination. Disruption of yeaZ did not affect cell growth significantly in 2216 E broth at 28°C. The wild-type and mutant viable but nonculturable (VBNC) cells could be resuscitated by temperature upshift method. In addition, the recombinant YeaZ increased the culturable counts from 1·27 × 10 4  CFU per ml and 1·99 × 10 4 CFU per ml to 2·88 × 10 5  CFU per ml and 4·59 × 10 5 CFU per ml, respectively. After the VBNC cells of wild-type and mutant cells were maintained at 4°C for 120 days, no resuscitation was obtained by temperature upshift method, but addition of the recombinant YeaZ promoted the resuscitation of the wild-type and mutant cells, with the culturable cell counts of 1·13 × 10 3 and 1·44 × 10 3 CFU per ml, respectively. Disruption of yeaZ decreased the virulence of V. harveyi in zebrafish. The lethal dose 50% of the yeaZ null mutant was more than 10-fold higher than that of the wild-type cells. The recombinant YeaZ could efficiently promote resuscitation of the wild-type and mutant cells of V. harveyi from VBNC to culturable state. The protein also promoted resuscitation of the VBNC wild-type and mutant cells, which were maintained at 4°C for 120 days and not recovered by temperature upshift method. Disruption of yeaZ decreased the virulence of V. harveyi in zebrafish. Here, we show clear evidence of a resuscitation-promoting factor YeaZ of V. harveyi and the roles in resuscitation of the VBNC cells and its pathogenicity. © 2016 The Society for Applied Microbiology.

  3. Pleurocidin Peptide Enhances Grouper Anti-Vibrio harveyi Immunity Elicited by Poly(lactide-co-glycolide-Encapsulated Recombinant Glyceraldehyde-3-phosphate Dehydrogenase

    Directory of Open Access Journals (Sweden)

    Shu-Chun Chuang

    2014-05-01

    Full Text Available Outer membrane proteins, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH, are considered immunodominant antigens for eliciting protective immunity against Vibrio harveyi, the main etiological agent of vibriosis in fish. Cationic antimicrobial peptides (AMPs, such as pleurocidin (PLE, play important roles in activating and recruiting immune cells, thereby contributing to subsequent innate and adaptive immune responses. In the present study, we aimed to use PLE peptide as a potent adjuvant to improve the immunogenicity of V. harveyi recombinant GAPDH (rGAPDH. In order to prepare a controlled-release vaccine, PLE peptide and rGAPDH protein were simultaneously encapsulated into polymeric microparticles made from the biodegradable poly(lactide-co-glycolide (PLG polymer. The resulting PLG-encapsulated PLE plus rGAPDH (PLG-PLE/rGAPDH microparticles, 3.21–6.27 μm in diameter, showed 72%–83% entrapment efficiency and durably released both PLE and rGAPDH for a long 30-day period. Following peritoneal immunization in grouper (Epinephelus coioides, PLG-PLE/rGAPDH microparticles resulted in significantly higher (p < 0.05, nested design long-lasting GAPDH-specific immunity (serum titers and lymphocyte proliferation than PLG-encapsulated rGAPDH (PLG-rGAPDH microparticles. After an experimental challenge of V. harveyi, PLG-PLE/rGAPDH microparticles conferred a high survival rate (85%, which was significantly higher (p < 0.05, chi-square test than that induced by PLG-rGAPDH microparticles (67%. In conclusion, PLE peptide exhibits an efficacious adjuvant effect to elicit not only improved immunity, but also enhanced protection against V. harveyi in grouper induced by rGAPDH protein encapsulated in PLG microparticles.

  4. Effect of probiotic supplemented diet on marine shrimp survival after challenge with Vibrio harveyi Efeito do uso de dieta suplementada com o probiótico sobre a sobrevivência de camarões marinhos após o desafio com Vibrio harveyi

    Directory of Open Access Journals (Sweden)

    F.N. Vieira

    2010-06-01

    Full Text Available The effect of a Lactobacillus plantarum-supplemented diet on shrimp growth, digestive tract bacterial microbiota, survival, and some hemato-immunological parameters after an experimental challenge with Vibrio harveyi was studied. No difference (P>0.05 was observed in shrimp survival (80±6% and final weight (6.63±0.56g after 60 days feeding trial. Total bacteria count and Vibrio spp. count in the digestive tract were not diferent (P>0.05 until day 40, but they were lower (P0.05 than the control group (39.9±4.4%. Bacterial counts in hemolymph and hepatopancreas was lower (P0.05 in the probiotic-supplemented group after challenge with V. harveyi. Probiotic-supplemented diet modifies shrimp digestive tract bacterial microbiota, increasing resistance to V. harveyi infection.Avaliou-se o efeito de uma dieta suplementada com Lactobacillus plantarum na engorda de Litopenaeus vannamei sobre a microbiota bacteriana do trato digestivo, a sobrevivência e alguns parâmetros hematoimunológicos após desafio com Vibrio harveyi. Não foi observada diferença significativa na sobrevivência (80±6% e no peso final dos camarões (6,63±0,56g durante a engorda (60 dias. A população de bactérias totais e Vibrio spp. no intestino não diferiu até o 40° dia de cultivo, mas foi inferior nos camarões alimentados com dieta suplementada com probiótico no 60° dia de cultivo. A população de bactérias lácticas foi superior no intestino de camarões alimentados com dieta suplementada com probióticos após o 20° dia de cultivo. Dez horas após o desafio, a sobrevivência dos camarões alimentados com dieta suplementada com probiótico foi superior (65,7±2,9% a dos camarões do controle (39,9±4,4%. Camarões alimentados com dieta suplementada com probióticos e desafiados com V. harveyi apresentaram menor contagem de bactérias na hemolinfa e hepatopâncreas. A contagem total de hemócitos e a atividade aglutinante da hemolinfa foram superiores nos camar

  5. Elevated cytokine responses to Vibrio harveyi infection in the Japanese pufferfish (Takifugu rubripes) treated with Lactobacillus paracasei spp. paracasei (06TCa22) isolated from the Mongolian dairy product.

    Science.gov (United States)

    Biswas, G; Korenaga, H; Nagamine, R; Kawahara, S; Takeda, S; Kikuchi, Y; Dashnyam, B; Yoshida, T; Kono, T; Sakai, M

    2013-09-01

    With the aim of evaluating the effect of a Mongolian dairy product derived Lactobacillus paracasei spp. paracasei (strain 06TCa22) (Lpp) on the cytokine-mediated immune responses to Vibrio harveyi infection, we examined 16 cytokine expressions in the Japanese pufferfish, Takifugu rubripes. Fish were orally treated with the heat-killed Lpp at 1 mg g(-1) body weight d(-1) for 3 days. At 24 h posttreatment, fish were infected by an intramuscular injection of 0.1 mL V. harveyi bacterial suspension (10(8) cfu mL(-1)). Additionally, superoxide anion production (SAP) and phagocytic activity (PA) of head kidney cells were assessed during 120 h postinfection period. Significant up-regulation of pro-inflammatory (IL-1β, IL-6, IL-17A/F-3, TNF-α and TNF-N), cell-mediated immune inducing (IL-12p35, IL-12p40 and IL-18), antiviral/intra-cellular pathogen killing (I-IFN-1 and IFN-γ), anti-inflammatory (IL-10) and lymphocyte agonistic (IL-2, IL-7, IL-15, IL-21 and TGF-β1) cytokines was observed in the treated fish compared to control ones during the pathogen infection. Furthermore, significantly increased SAP and PA (P < 0.01; 0.05) were recorded in the treated fish compared to untreated fish. These results suggest the beneficial role of Lpp in enhancement of cytokine-mediated immunity in the Japanese pufferfish against V. harveyi infection and application of this product as a potential fish immunostimulant. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Control of Vibrio harveyi Infection in Blue Swimming Crab, Portunus pelagicus Larvae by the Gut Isolated Lactic Acid Bacteria under Challenge Bioassay

    Directory of Open Access Journals (Sweden)

    Allah Dad Talpur*§, Abdul Jabbar Memon§, Muhammad Iqbal Khan§, Muhammad Ikhwanuddin, Muhammad Mhd Danish Daniel2 and Ambok Bolong Abol-Munafi

    2012-06-01

    Full Text Available Five isolates of lactic acid bacteria (LAB isolated from the gut of female Portunus pelagicus with inhibitory activity against shellfish pathogens and validation as probiotics via small scale in vivo model were tested for hatchery trials pathogen probiotic assay. Vibrio harveyi previously isolated from the gut of P. pelagicus, was added at 104 cfu mL-1 to test larvae for 10h. Test (LAB isolates were inoculated at a concentration of 106 cfu mL-1 to pathogen addition aquaria once and until day four during the experiment. 20 larvae/liter were stocked and larval survival was determined over five days. Lactobacillus plantarum did produce highest survival 28.33% to 48.33% in one day and daily inoculations respectively and in probiotic control it produced 58.33% survival followed by L. rhamnosus 55% and L. salivarius 53.33% respectively over non inoculated control 43.33% and 0% survival in V. harveyi inoculated control. However, Weissella confusa and W. cibaria did show less probiotic activity compared to rests of three LAB isolates. In the present study, it was determined that three LAB probiotics were effective in hatchery trials challenge assays which may significantly control the infection and increased the survival of larvae.

  7. Effect of alkaloids derived from jellyfish (Aeginura sp.) on the intestinal histopathology and relative percentage survival (RPS) of tiger grouper (Epinephelus fuscoguttatus) infected by Vibrio harveyi

    Science.gov (United States)

    Andayani, S.; Fajar, M.; Rahman, M. F.

    2018-04-01

    The purposes of this research were to determine the effect of alkaloid jellyfish compounds on intestinal histopathology of tiger grouper and to determine the best doses to the relative percent survival (RPS) of tiger grouper. The method of this research was descriptive with completely randomized design. The treatment of active alkaloid compound on feed was investigated for 28 days. The fish were then challenged with Vibrio harveyi at 105 CFU/cell for 7 days. Alkaloids were added to the feed with the doses (g alkaloid/kg feed) of 0 (control); A = 0.5; B = 0.75; C = 1.0; and D = 1.25. The intestinal histopathology and RPS were observed. The best RPS was found at a treatment of C with the value of 100 %.

  8. Genes encoding the Vibrio harveyi haemolysin (VHH)/thermolabile haemolysin (TLH) are widespread in Vibrios%VHH/TLH溶血素基因在海洋弧菌中分布的研究

    Institute of Scientific and Technical Information of China (English)

    王淑娴; 张晓华; 钟英斌; 孙铂光; 陈吉祥

    2007-01-01

    deduced amino acid sequences are up to 85.6%), is a putative virulence factor to marine cultured fish. A VHH probe, which is specific to V. harveyi vhhA haemolysin gene, was used to screen EcoR Ⅰ digests of total DNA from 57 vibrio strains, including 26 vibrio type strains, 20 V. harveyi isolates and 11 V. parahaemolyticus isolates. As a result, 1 strong hybridisation band was detected in 13 type strains, including 2 of Vibrio alginolyticus , 2 of V. harveyi , and 1 strain each of Grimontia hollisae , V. campbellii , V. cincinnatiensis , V. fischeri , V. mimicus , V.natriegens, V. parahaemolyticus, V. proteolyticus and V. logei. Also, 1 weak band was detected in 6 type strains, including V.anguillarum, V. aestuarianus, Photobacterium damselae subsp. damselae, V. fluvialis, V. furnissii and V. vulnificus. There was not any hybridization signal in other type strains. Also, vhh/tlh was present in all isolates of V. harveyi and V. parahaemolyticus. Moreover, 3 isolates of V. harveyi, i.e. VIB 645, VIB 648 and SF1, had duplicated vhh genes. The data indicates that vhh/tlh is widespread in vibrios,especially in V. harveyi related species and V. fischeri related species. To support this conclusion, the vhh/tlh homologue genes in V.anguillarum VIB 72, V. campbellii VIB 285, V. natriegens VIB 299 and V. harveyi VIB 647 were cloned and sequenced, and the deduced amino acid sequences showed high degree of identities to VHH (67% ~ 99% ) and TLH haemolysin (69% ~ 91% ). This study will help us to identify the role of vhh/tlh haemolysin gene in the pathogenicity of vibrios.

  9. Promoter Boundaries for the luxCDABE and betIBA-proXWV Operons in Vibrio harveyi Defined by the Method Rapid Arbitrary PCR Insertion Libraries (RAIL).

    Science.gov (United States)

    Hustmyer, Christine M; Simpson, Chelsea A; Olney, Stephen G; Rusch, Douglas B; Bochman, Matthew L; van Kessel, Julia C

    2018-06-01

    Experimental studies of transcriptional regulation in bacteria require the ability to precisely measure changes in gene expression, often accomplished through the use of reporter genes. However, the boundaries of promoter sequences required for transcription are often unknown, thus complicating the construction of reporters and genetic analysis of transcriptional regulation. Here, we analyze reporter libraries to define the promoter boundaries of the luxCDABE bioluminescence operon and the betIBA-proXWV osmotic stress operon in Vibrio harveyi We describe a new method called r apid a rbitrary PCR i nsertion l ibraries (RAIL) that combines the power of arbitrary PCR and isothermal DNA assembly to rapidly clone promoter fragments of various lengths upstream of reporter genes to generate large libraries. To demonstrate the versatility and efficiency of RAIL, we analyzed the promoters driving expression of the luxCDABE and betIBA-proXWV operons and created libraries of DNA fragments from these loci fused to fluorescent reporters. Using flow cytometry sorting and deep sequencing, we identified the DNA regions necessary and sufficient for maximum gene expression for each promoter. These analyses uncovered previously unknown regulatory sequences and validated known transcription factor binding sites. We applied this high-throughput method to gfp , mCherry , and lacZ reporters and multiple promoters in V. harveyi We anticipate that the RAIL method will be easily applicable to other model systems for genetic, molecular, and cell biological applications. IMPORTANCE Gene reporter constructs have long been essential tools for studying gene regulation in bacteria, particularly following the recent advent of fluorescent gene reporters. We developed a new method that enables efficient construction of promoter fusions to reporter genes to study gene regulation. We demonstrate the versatility of this technique in the model bacterium Vibrio harveyi by constructing promoter libraries

  10. Effects of dietary hawthorn extract on growth performance, immune responses, growth- and immune-related genes expression of juvenile golden pompano (Trachinotus ovatus) and its susceptibility to Vibrio harveyi infection.

    Science.gov (United States)

    Tan, Xiaohong; Sun, Zhenzhu; Huang, Zhong; Zhou, Chuanpeng; Lin, Heizhao; Tan, Lianjie; Xun, Pengwei; Huang, Qian

    2017-11-01

    The present study was conducted to investigate the effects of dietary hawthorn extract (HTE) supplementation on growth performance, immune responses, hepatic antioxidant abilities, growth- and immune-related and heat shock protein genes expression and resistance to the pathogen Vibrio harveyi in Trachinotus ovatus. A basal diet supplemented with HTE at 0 (Diet 1), 0.50 (Diet 2), 1.00 (Diet 3), 2.00 (Diet 4), 4.00 (Diet 5) and 10.00 (Diet 6) g kg -1 were fed to golden pompano for 8 weeks. The highest final body weight, weight gain rate, specific growth rate, feed efficiency ratio and protein efficiency rate were observed in fish fed Diet 2 (P Vibrio harveyi, significant higher post-challenge survival was observed in fish fed Diet 2 and Diet 3 than the control group (P growth-related genes (IGF-I and IGF-II) were significantly up-regulated in fish fed HTE supplement (P growth performance and growth-related genes expression, strengthen immunity, and improve hepatic antioxidative abilities and resistance to Vibrio harveyi infection. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Screening and Identification of Anti-Vibrio harveyi Lactic Acid Bacteria Derived from Marine Fish Intestine%海水鱼肠道源性抗哈维氏弧菌乳酸菌的筛选与鉴定

    Institute of Scientific and Technical Information of China (English)

    马欢欢; 吕欣然; 林洋; 白凤翎; 励建荣

    2017-01-01

    Vibrio harveyi is a main pathogenic bacterium that can induce vibriosis of shrimp and fish in the marine environment,causing huge economic losses in aquaculture.In this paper,49 strains of lactic acid bacteria (LAB) were isolated from the marine fish intestine by the use of 1.0% CaCO3 MRS agar,including seabass,coilia ectenes,Paralichthys lethostigma and turbot.strain YP4-5 isolated from the intestine of Paralichthys lethostigma had strong antagonistic activity against V.harveyi using the method of Oxford cup agar diffusion,showing an inhibitory zone diameter of 18.26 mm.Antimicrobial substances of the cell-free supernatant (CFS) produced by strain YP4-5 were sensitive to protease,heat stable and effective within the pH range of 2.5-4.5,and they were preliminary determined as bacteriocins.The growth curve indicated that 74.74% of V.harveyi was inhibited by treatment with the CFS at a concentration of 0.4 MIC.Furthermore,cell integrity damage,cell membrane dissolution and intracellular component leakage were observed by scanning electron microscope.Strain YP4-5 was identified as Lactobacillus sakei according to physiological and biochemical characteristics and 16S rRNA sequence analysis.This research can provide a good source of probiotics to prevent and control V.harveyi infection in aquaculture.%哈维氏弧菌是引起对虾和鱼弧菌病的主要病原菌,给海产养殖业造成重大的经济损失.利用1.0% CaCO3MRS琼脂从鲈鱼、刀鱼、牙鲆和大菱鲆等海水鱼肠道中分离到49株乳酸菌,采用牛津杯琼脂扩散法从中筛选出对哈维氏弧菌具有较强拮抗作用的菌株YP4-5,抑菌直径为18.26 mm.通过蛋白酶、pH值和温度等因素对菌株YP4-5的无细胞上清液(cell free supematant,CFS)抑菌物质进行初步分析,表明抑菌活性物质对蛋白酶敏感,pH2.5~4.5时对哈维氏弧菌具有抑制作用,具有良好的热稳定性,初步判断YP4-5 CFS中抑菌活性物质为细菌素类.0.4

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

    International Nuclear Information System (INIS)

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

    2009-01-01

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

  13. Expression of Vibrio harveyi acyl-ACP synthetase allows efficient entry of exogenous fatty acids into the Escherichia coli fatty acid and lipid A synthetic pathways.

    Science.gov (United States)

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

    2010-02-02

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

  14. Growth, nonspecific immune characteristics, and survival upon challenge with Vibrio harveyi in Pacific white shrimp (Litopenaeus vannamei) raised on diets containing algal meal.

    Science.gov (United States)

    Nonwachai, Thasanee; Purivirojkul, Watchariya; Limsuwan, Chalor; Chuchird, Niti; Velasco, Mario; Dhar, Arun K

    2010-08-01

    A 70-day growth trial was conducted with postlarvae 12 (PL12) Pacific white shrimp (Litopenaeus vannamei) to study the suitability of soybean meal and oil originating from a single-celled microorganism (thraustochytrid) as fishmeal and fish oil substitutes in practical diets for L. vannamei. The growth, survival rate and immune characteristics were evaluated. Seven experimental diets were designed with soybean meal used as the primary protein source; each formulation contained 33% crude protein and 8% lipid. Fish oil was completely substituted with 3% soybean oil and meals originating from single-celled heterotrophs rich in docosahexaenoic acid (DHA) and arachidonic acid (ARA) were added at different concentrations. A commercial shrimp feed was used as the control diet. The final weights and survival rates of the shrimp were not significantly different among all treatments. However, shrimp raised on diets supplemented with marine algal meals rich in DHA and ARA showed significant improvement in immune parameters, such as total hemocyte count, phenoloxidase activity, superoxide dismutase activity, and bactericidal activity. Additionally, the survival rate after challenge with Vibrio harveyi was increased. These findings demonstrated that substitution of thraustochytrid-derived meals as an alternative to fish-based ingredients in shrimp diets provided similar growth rates while increasing the immune parameters and providing vibriosis resistance. Copyright 2010 Elsevier Ltd. All rights reserved.

  15. 哈氏弧菌EcGY020401优化培养%Optimization Culture of Vibrio harveyi EcGY020401

    Institute of Scientific and Technical Information of China (English)

    陶家发; 赖迎迢; 潘厚军; 石存斌; 吴淑勤

    2010-01-01

    目的:为规模化制备弧菌疫苗提供相关数据.方法:利用摇瓶和小型发酵罐培养,通过平板计数测定培养菌液的活菌数.结果:哈氏弧菌(Vibrio harveyi)EcGY020401株最适盐度为20~25g/L,合适的pH为7.5~7.7,葡萄糖浓度最适浓度为2~5g/L,用TSB优化培养基:胰蛋白胨5g/L、蛋白胨15g/L、大豆蛋白胨3g/L、酵母膏1g/L、葡萄糖4g/L、磷酸氢二钾5g/L、氯化钠15g/L、pH 7.5,培养EcGY020401菌株,可达2.95×1010 cfu/mL;小型发酵罐培养,10h可达到生长最大值,菌液浓度为3.46×1010cfu/mL 结论:用优化TSB培养基,采用发酵罐培养弧菌,可降低成本,提搞培养效率.

  16. 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.

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

    Science.gov (United States)

    Pholdaeng, Komsil; Pongsamart, Sunanta

    2010-04-01

    Oral administration of polysaccharide gel (PG) in shrimp diets revealed immunostimulating potential and disease resistance in Penaeus monodon (black tiger shrimp). PG from the fruit-rind of Durio zibethinus has been characterized to be a pectic polysaccharide with immunomodulating and antibacterial activities. PG inhibited growth of the shrimp bacterial pathogen, Vibrio harveyi 1526, by agar diffusion and broth microdilution tests. Clear inhibition zones on agar plates were observed at the lowest PG concentration of 3.1 mg/ml, where minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values for PG were 6.3 and 12.5 mg/ml, respectively. Each group of juvenile shrimps, initial mean body weight 0.29 +/- 0.04 g, was housed in a closed-recirculating treated water system and was fed with PG-supplemented diets containing 1, 2 and 3% PG or shrimp basal diet in the control group for 8 and 12 weeks. PG-supplemented diets did not contribute to the overall growth of black tiger shrimp. The immune response was evaluated by analysis of prophenoloxidase activity and total hemocyte count in the shrimp fed PG-supplemented diets for 12 weeks. Prophenoloxidase activity in shrimp fed the 1, 2 and 3% PG-supplemented diet and total hemocyte count in shrimp fed the 1 and 2% PG-supplemented diet were higher (P < 0.05) than those of the control group. The percent survival was higher in groups fed the 1-3% PG-supplemented diets in challenge tests with either white spot syndrome virus (WSSV) or the bacterium V. harveyi 1526 than that of the control group. Relative percent survival (RPS) values in groups fed the 2% PG-supplemented diet showed the highest RPS value for disease resistance of 100% (at Day 6) and 36% (at Day 4) in treated shrimp against viral and bacterial infection, respectively. Mortality of PG-supplemented diets in treated shrimps against WSSV infection was also found to be much lower (P < 0.05) than that of the control group. Copyright 2009

  18. Avaliação de bacterina e Lactobacillus plantarum frente à infecção experimental por Vibrio harveyi em pós-larvas de Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Celso Carlos Buglione

    2008-12-01

    Full Text Available This study aimed to verify the effect of probiotics and inactivated cells of bacterias such as Vibrio alginolyticus, Aeromonas salmonicida and Pasteurella multocida in larvae survival of Litopenaeus vannamei, in stress test and experimental infection with Vibrio harveyi. Conic tanks of 30 L, were stocked with 400 post-larvae stage five. Four experimental treatments with triplicates consisted of: 1: commercial feed (control, 2: commercial feed plus bacterin by oral administration in artemia, 3: commercial feed plus bacterin by immersion administration, 4: commercial feed with Lactobacillus plantarum inoculation. Bacterin application was conducted 6h before the infection and stress test, while probiotic administration was for 15 days before challenges. In stress test, post-larvae of treatment 4 (commercial feed supplemented with Lactobacillus plantarum with reached the highest survival rate (87,86 ± 2,35% followed by the ones of treatment 3 and 2 (bacterim by immersion and bacterim by oral administration in artemia with 81,54±1,50% and 80,16 ± 2,15%, respectively, which were superior to the control treatment (72,63 ± 3,34%. Next to V. harveyi challenge, animals from treatment 3 presented the highest survival rate (79,60 ± 7,12% followed by treatments 4 (69,60 ± 10,43%, 2 (65,60 ± 5,18% and control (56,4 ± 5,58%. All treatments were different from control. The present results demonstrate the possible use of L. plantarum and bacterin as promoters in survival rates of L. vannamei post-larvae in the stress tests and challenges with Vibrio harveyi.

  19. 黄芩乙醇提取物通过下调NAD特异的谷氨酸脱氢酶抑制哈维氏弧菌生长%Ethanol-extracts from Scutellaria Inhibit the Growth of Vibrio harveyi by Downregulating NAD +-dependent Glutamate Dehydrogenase

    Institute of Scientific and Technical Information of China (English)

    谢丽玲; 朱琳; 王爱霞; 朱炎坤; 黎家杰; 周亮; 毕潇

    2017-01-01

    Vibrio harveyi is one of common pathogenic bacteria,which is seriously imperilling human health and aquatic industry.Because of antibiotics abuse,drug residues and drug-resistance are becoming more serious.It is urgent to look for new alternatives with low toxicity and susceptiblihy to drug-resistance.In this study,we reveal the antibacterial effects of the ethanol extracts from Scutellaria on V.harveyi.The results showed that the ethanol extracts of Scutellaria had a strong inhibitory effect on V.harvey,the diameter of inhibition zone was 18.33 ± 0.58 mm.The minimum inhibitory concentrations (MIC) and minimal bactericidal concentration (MBC) were 7.92 mg/mL and 15.84 mg/mL,respectively.By observation with scanning electron microscope (SEM) and concentration determination of intracellular and extracellular proteins,we found that although there were many little pores on the surface of V.harveyi,it remained smooth and integrated.After treated with ethanol extracts of Scutellaria,a band of protein disappeared in SDS-PAGE which was identified as NAD-specific glutamate dehydrogenase (NAD-GDH) by MALDI-TOF-TOF-MS.Furthermore,the real-time PCR results showed that the mRNA levels of this gene were affected by ethanol extracts of Scutellaria.These results are in good agreement with Scutellaria application in aqueous infection,and indicate that ethanol extracts of Scutellaria inhibit V.harveyi growth effectively through downregulating the expression of NAD-dependent glutamate dehydrogenase,which provides new evidence for the application of traditional Chinese medicine in the fields of aquaculture.%哈维氏弧菌(Vibrio harveyi)是水产动物的常见致病菌,对人类健康和水产经济带来巨大威 胁.抗生素的滥用使得药物残留和耐药性问题变得日益严重.因此,迫切需要寻找新型、不易产生耐药性和低毒的抗菌物质.本文研究黄芩醇提物对哈维氏弧菌的抑制作用及抑菌机制.实验结果表明,黄芩醇提物对

  20. Investigation of Vibrio alginolyticus, V. harveyi, and V. parahaemolyticus in large yellow croaker, Pseudosciaena crocea (Richardson) reared in Xiangshan Bay, China

    OpenAIRE

    Lu Liu; Mingfeng Ge; Xiaoye Zheng; Zhen Tao; Suming Zhou; Guoliang Wang

    2016-01-01

    Large yellow croaker (LYC), Pseudosciaena crocea is an economically important fish species of mariculture in China. The variation of yearly production of LYC has been increasingly related to the outbreaks of fish diseases. Moreover, Vibrio infections have been identified in this fish frequently. To understand the pattern of Vibrio infections in LYC, we conducted a culture-independent survey of Vibrios in farmed LYC populations using a multiplex PCR method targeting Vibrio alginolyticus, Vibri...

  1. Leucaena leucocephala pod seed protein as an alternate to animal protein in fish feed and evaluation of its role to fight against infection caused by Vibrio harveyi and Pseudomonas aeruginosa.

    Science.gov (United States)

    Verma, Vipin Kumar; Rani, Kumari Vandana; Kumar, Shiva Raj; Prakash, Om

    2018-05-01

    The laboratory acclimatized Clarias gariepinus (80 ± 10 g) were divided into six groups and five subgroups each containing 10 fish. A fish feed was reconstituted by adding 33% powder of Leucaena leucocephala seed in place of fish trash. Group B, C and E were fed on reconstituted feed and group A, D and F were fed on artificial feed containing animal protein for 7 days prior to start of experiments. Then Group B was challenged with BSA while other groups were challenged with Vibrio harveyi (Group C, D) and Pseudomonas aeruginosa (Group E, F). Group A was used as negative control (not challenged with antigen). The fish were challenged on weekly intervals till 28th day. Blood was collected from one subgroup of each group on day 7, 14, 21 & 28 and finally sacrificed on day 35. Change in body weight, liver function tests (SGOT, SGPT) and serum ALP levels were monitored. The phagocytic index, percentage phagocytosis and nitric oxide levels were measured in macrophages isolated from spleen and head kidney. The levels of total fish immunoglobulin were also measured following indirect ELISA. The results showed improved immune response in fish fed on 33% L. leucocephala pod seed reconstituted feed; however their specific growth rate was low. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. Antibiogram types of Vibrio harveyi strains isolated from maricultured fish%海水养殖鱼哈维弧菌分离株的耐药谱型分析

    Institute of Scientific and Technical Information of China (English)

    曾德乾; 冯娟; 徐力文; 苏友禄; 郭志勋

    2015-01-01

    The present study investigated the antibiogram types of Vibrio harveyi isolates collected from diseased fish that were cultured along the south China coast. The antibiotic resistance profiles of 84 strains were tested with 12 common antibiotics using Kirby–Bauer disc diffusion methodology. The antimicrobial susceptibility data were used to identify correlations between antibiogram and subgroup types using cluster analysis software (SPSS 19.0, Statistical Product and Service Solutions). Eighty-four V. harveyi isolates formed 26 antibiogram types with an antibiogram abundance of 31.0%. The number of multiple antibiotic resistant types (i.e., resistant to more than three antibiotics) in the strains was 13 and comprised 50.0% of the total antibiogram types, causing the multi-antibiogram abundance value to increase to 59.1%. Isolates originating from Hainan, Guangdong, Guangxi, and Fujian Provinces possessed 18, 15, 3, and 1 of the antibiogram types, respectively, and all shared the J antibiogram type (FUR/AMO). Representative antibiogram types were J and M from 2007, A and B were added in 2010, 24 A to X types (A to X except for Y, Z) were added in 2011, while Y and Z appeared in 2012. The kinds of suppressive antimicrobial agents was 2, 5, 7, and 8, thereby mirroring the sampling years, 2007, 2010, 2011 and 2012, respectively. Strains isolated from Epinephelus awoara, E. fuscoguttatus, Plectropomus leopardus, Trachinotus ovatus and E. lanceolatus♂ × E. fuscoguttatus♀ contained 6–10 antibiogram types, while the remaining fish sampled had only 1–3 types. The strains clustered initially into six subgroups (i–vi) and further formed groups I and II. Each subgroup contained characteristic antibiogram types comprising N, P, T, U, Y;R, S;K, P; F, H, O, X, V, W; E, G, Q; and C, L, Z. Common antibiogram types were not found in the Group I and II strains. Our research revealed that the V. harveyi isolates displayed polymorphic antibiogram types of moderately high

  3. POTENSI BEBERAPA ISOLAT PROBIOTIK SEBAGAI ANTIBAKTERI TERHADAP PERTUMBUHAN Vibrio spp.

    OpenAIRE

    HASBIAH

    2015-01-01

    The research about potential of some probiotic isolates as an antibacterial on the growth of Vibrio spp had been done. This research aimed to know the antibacterial potency from some isolates probiotic on the growth of Vibrio spp. This research to tested the inhibition on the three species of Vibrio that are Vibrio harveyi, Vibrio prahaemolyticus, and Vibrio cholerae using agar diffusion method. Probiotic isolates come from lactic acid bacteria group that provide beneficial effects on health ...

  4. Preliminary Study on One Pathogenic Bacterium---Vibrio harveyi Associated with Red Body Disease in Cultured Exopalaemon carinicauda%一种引起脊尾白虾红体病病原菌的初步研究

    Institute of Scientific and Technical Information of China (English)

    张文文; 王庚申; 施慧; 谢建军; 许文军

    2014-01-01

    In July 2012, a serious pathogentic disease outbroke and quickly spread in Exopalaemon carinicauda which were temporary bred in indoor farm in Zhoushan of Zhejiang province. Incidence rates were around 30% in a wee k (the highest mortality rate of 0.5%). Diseased shrimp showed typical red body disease characteristics: systemic red bodies, unresponsive, feeding reduced or even stopt, swollen liver, pancreas and empty sotmach, etc. A bacterial strain named XS1207005 was isolated from the diseased E. carinicauda. In ar-tificial infection test, XS1207005 was proved to be the pathogen. Amplification and sequencing of the 16S rRNA and HSP60 gene followed by phylogenetic analysis confirmed its identity. According to the traditional biochemical identification and 16S rRNA and HSP60 homology analysis, the pathogenic bacterium was V. har-veyi. Phylogenetic tree of Vibrios based on HSP60 gene sequences revealed that strain XS1207005 showed the highest level of similarity to V. harveyi (EU036994, AF230934, AY332569). HSP60 gene sequences showed that strain XS1207005 had sequence identity of more than 99.6% to V. harveyi. The results of sensitivity to 22 kinds of antibacterial agents showed that strain XS1207005 was sensitive to cefotaxime, sulfamethoxazolum, ri-fampicine, cefoperazone, ceftriaxone, norfloxacin, ceftazidime, ofloxacin. V. harveyi is a kind of pathogenic bacterium commonly found in marine-fishes culture, but this is the first time that V. harveyi is reported as the pathogenic bacteria of cultured E. carinicauda in China.%2012年7月,浙江舟山某试验场暂养的脊尾白虾暴发一种较为严重的传染性疾病,病虾身体发红,额剑发黑,反应迟钝,摄食减少,一周内发病死亡率达到30%左右,并有迅速蔓延趋势。从病虾的血淋巴液和肝胰腺中分离到一株优势细菌XS1207005。人工感染试验结果表明,该菌株对健康脊尾白虾有较强的致病性,且感染发病的脊尾白虾与自然发

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

    Science.gov (United States)

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

    2014-01-01

    In this study, vegetative cell suspensions of two Bacillus subtilis strains, L10 and G1 in equal proportions, was administered at two different doses 10(5) (BM5) and 10(8) (BM8) CFU ml(-1) in the rearing water of shrimp (Litopenaeus vannamei) for eight weeks. Both probiotic groups showed a significant reduction of ammonia, nitrite and nitrate ions under in vitro and in vivo conditions. In comparison to untreated control group, final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR) and digestive enzymatic activity were significantly greater in the BM5 and BM8 groups. Significant differences for survival were recorded in the BM8 group as compared to the control. Eight weeks after the start of experiment, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 80%, whereas cumulative mortality of the shrimp that had been given probiotics was 36.7% with MB8 and 50% with MB5. Subsequently, real-time RT-PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and β-1,3-glucan- binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was only significantly up-regulated in the BM5 group compared to the BM8 and control groups. These results suggest that administration of B. subtilis strains in the rearing water confers beneficial effects for shrimp aquaculture, considering water quality, growth performance, digestive enzymatic activity, immune response and disease resistance. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Screening of probiotic bacteria and its role on artificial infection of Vibrio harvey in white shrimp (Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    . Sukenda

    2007-07-01

    Full Text Available Probiotic was screened from 28 strains of normal bacterial flora isolated from rearing water in a Litopenaeus vannamei farm based on its inhibitory activity against the growth of Vibrio harveyi.  Antibacterial activity was also tested in vivo to V. harveyi in L. vannamei.  The result showed that  the probiotic has a antibacterial effect on V. harveyi.  The in vivo test showed that shrimps injected with probiotic previously before challenged with V. harveyi has survival higher than control.  Probiotic isolate was suspected as Vibrio furnissi. Keywords:  biocontrol, inhibitory activity, Vibrio furnissi, Vibrio harveyi, Litopenaeus vannamei   AbstraK Bakteri probiotik ditapis dari 28 strain bacteria flora yang diisolasi dari air pemeliharaan udang vaname Litopenaeus vannamei berdasarkan aktivitas penghambatannya terhadap pertumbuhan Vibrio harveyi.  Aktivitas bakteri probiotik juga diuji secara in vivo terhadap V. harveyi pada udang putih.  Hasil penelitian menunjukkan bahwa bakteri probiotik isolat memiliki kemampuan antibakteria terhadap V. harveyi.  Uji in vivo menunjukkan bahwa udang yang diinjeksi probiotik sebelum diuji tantang dengan V. harveyi memiliki kelangsungan hidup lebih tinggi daripada kontrol.  Isolat probiotik tersebut diduga adalah Vibrio furnissi. Kata kunci:  biokontrol, aktivitas penghambatan, Vibrio furnissi, Vibrio harveyi, Litopenaeus vannamei

  7. 大蒜对印度明对虾黑鳃病原耐药菌株哈维氏弧菌抗菌活性的研究%Antibacterial activity of Allium sativum against multidrug-resistant Vibrio harveyi isolated from black gill-diseased Fenneropenaeus indicus

    Institute of Scientific and Technical Information of China (English)

    杨仲明(译)

    2012-01-01

    本研究针对大蒜提取物对印度明对虾(Fenneropenaeusindicus)黑腮病病原多重耐药哈维氏弧菌(Vibrioharveyi)体外和体内的抗菌活性进行了评价。先利用市售的几种抗生素记录致病性哈维氏弧菌的抗菌谱。然后,利用琼脂扩散法来研究大蒜对哈维氏弧菌的抗菌活性。将哈维氏弧菌接种在琼脂平板上,对每种大蒜提取物(鲜榨大蒜提取物-FsGE,冻干大蒜提取物-FDGE以及用甲醇提取的大蒜提取物-MGE)设计三种不同的浓度。结果表明,鲜榨大蒜提取物对哈维氏弧菌的抑菌活性明显优于冻干大蒜提取物和用甲醇提取的大蒜提取物。本研究分别在对虾基础饲料中添加O%(对照)、0.5%和1%浓度梯度的大蒜来评价大蒜治疗感染哈维氏弧菌的对虾的效果。结果表明,综合大蒜提取物的短期和长期防治(1%含量)的效果分析,对虾累积死亡率降低了75%。本研究探讨了鲜榨大蒜提取物替代抗生素在印度明对虾饲养防病上的优点。%The present study evaluated the in vitro and in vivo inhibitory effects of Allium sativum extracts against the multidrug- resistant Vibrio harveyi, isolated from the black gill-diseased Fenneropenaeus indicus. Initially, the antibiogram of pathogenic V. harveyi was documented for its resistance towards selected commercial antibiotics. Further, antibacterial activity of A. sativum against Vibrio harveyi was performed by the agar well diffusion method. V. harveyi culture was seeded over the agar plates, and each of the garlic extracts (freshly squeezed garlic extract-FSGE, freeze-dried garlic extract-FDGE and methanolic garlic extract-MGE) was tested at three different concentrations. FSGE showed greater inhibitory activity against V. harveyi than FDGE and MGE. Three diets of standard meal preparation for shrimp containing 0% garlic as a control, 0. 5% garlic and 1% garlic treatments were formulated to

  8. 环介导等温扩增联合横向流动试纸条可视化检测哈维氏弧菌的研究%Visual Detection ofVibrio harveyi Based on Loop-mediated Isothermal Amplification Combined with a Lateral Flow Dipstick

    Institute of Scientific and Technical Information of China (English)

    程蝶; 柴方超; 蔡怡; 周前进; 陈炯

    2016-01-01

    Based on nucleotide enrichment by a loop-mediated isothermal amplification(LAMP)and chromatographic visualization by a lateral flow dipstick(LFD)assay,this work aims to develop a novel LAMP-LFD method for the rapid detection of Vibrio harveyi. Three pairs of primers were designed using the hemolysin gene(vhhA)of V. harveyi as detection target,and used in LAMP reaction,among which the forward inner primer vhhA-FIP was biotinylated. Similarly,a fluorescein isothiocyanate(FITC)-labeled probe vhhA-HP was designed to specifically hybridize with LAMP products. And then the hybridized LAMP products were visually detected by LFD. The optimized LAMP was performed at 63℃ for 40 min;and visual detection via LFD took 50 min. The results indicated that LAMP-LFD was able to specifically identify V. harveyi from other 9 pathogenic bacteria commonly existing in the aquatic animals,such as V. vulnificus. The detection limit of LAMP-LFD was 1.0×102 CFU/mL for V. harveyi pure cultures(equivalent to 2 CFU per reaction),and 5×102 CFU/mL for V. harveyi contaminated tissues of large yellow croaker(equivalent to 20 CFU per reaction),both of which were 100 times lower than that of the conventional PCR method using both outer primers vhhA-F3/vhhA-B3. Therefore,this rapid and accurate LAMP-LFD method is a promising alternative in the surveillance and point-of-care test of V. harveyiin sea farming.%以哈维氏弧菌(Vibrio harveyi)为材料,利用环介导等温扩增技术(LAMP)进行核酸扩增,借助横向流动试纸条(LFD)完成产物检测,旨在建立一种可用于哈维氏弧菌快速检测的LAMP-LFD新技术。以哈维氏弧菌的溶血素基因(vhhA)为检测靶标设计了3对特异性引物(其中,上游内引物vhhA-FIP由生物素标记),进行由生物素标记的LAMP反应;同时设计1条异硫氰酸荧光素(FITC)标记的探针,与获得的LAMP产物进行特异性杂交,杂交产物经LFD完成检测。经

  9. In vitro and in vivo effect of 2,6-Di-tert-butyl-4-methylphenol as an antibiofilm agent against quorum sensing mediated biofilm formation of Vibrio spp.

    Science.gov (United States)

    Santhakumari, Sivasubramanian; Jayakumar, Rengarajan; Logalakshmi, Ravichandran; Prabhu, Narayanan Marimuthu; Abdul Nazar, Abdul Kuthus; Karutha Pandian, Shunmugiah; Veera Ravi, Arumugam

    2018-05-25

    This study unveils the in vitro and in vivo antibiofilm potential of 2,6-Di-tert-butyl-4-methylphenol (DTBMP) from Chroococcus turgidus against Vibrio spp. In the preliminary study, cell free culture supernatant (CFCS) of C. turgidus inhibited the violacein production in biomarker strain Chromobacterium violaceum and its mutant strain CV026 in a dose dependent manner. The effective biofilm inhibitory concentration (BIC) of pure compound DTBMP from C. turgidus was identified as 250 μg/ml concentration in tested Vibrio species. Furthermore, DTBMP proved to effectively inhibit the bioluminescence production in V. harveyi and other biofilm related virulence traits such as exopolysaccharides (EPS) production, hydrophobicity index, swimming and swarming motility at its BIC concentration in three major pathogenic vibrios: V. harveyi, V. parahaemolyticus and V. vulnificus. The antibiofilm potential of DTBMP was validated through light, confocal laser scanning and scanning electron microscopic analyses. In addition, the non-bactericidal effect of DTBMP was determined through growth curve and 2,3-bis (2-methyloxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay. Real-time PCR studies revealed the down-regulation of master quorum sensing (QS) regulator genes of V. harveyi such as luxR, luxS, luxP, luxQ and luxO on treatment with DTBMP. In vivo results confirmed that DTBMP augmented the survival rate of Litopenaeus vannamei larvae up to 75, 88 and 66% upon infection with V. harveyi, V. parahaemolyticus and V. vulnificus, respectively. The results of this study ascertain the promising effects of DTBMP as an antibiofilm agent, which could be positively explored to treat biofilm-associated vibrios infections in aquaculture. Copyright © 2018 Elsevier B.V. All rights reserved.

  10. Screening of probiotic bacteria and its role on artificial infection of Vibrio harvey in white shrimp (Litopenaeus vannamei)

    OpenAIRE

    . Sukenda; A.J. Sihombing; Fitria Novianti; . Widanarni

    2007-01-01

    Probiotic was screened from 28 strains of normal bacterial flora isolated from rearing water in a Litopenaeus vannamei farm based on its inhibitory activity against the growth of Vibrio harveyi.  Antibacterial activity was also tested in vivo to V. harveyi in L. vannamei.  The result showed that  the probiotic has a antibacterial effect on V. harveyi.  The in vivo test showed that shrimps injected with probiotic previously before challenged with V. harveyi has survival higher than control.  P...

  11. Characterization of Vibrio species isolated from freshwater fishes by ribotyping

    OpenAIRE

    Mishra, P.; Samanta, M.; Mohanty, S.; Maiti, N. K.

    2010-01-01

    Three Vibrio species from the resident microflora of gastrointestinal tract of freshwater carps and prawns were isolated and confirmed biochemically as V. fluvialis from Cyprinus carpio/Labeo rohita; V. parahaemolyticus from Macrobrachium rosenbergii and V. harveyi from Macrobrachium malcomsoni. The genetic relationship among these Vibrio species was carried out by polymerase chain reaction (PCR) amplification of 16S rRNA gene followed by restriction digestion with Hae III, Bam HI and Pst I. ...

  12. Genomic taxonomy of vibrios

    Directory of Open Access Journals (Sweden)

    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

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

    Science.gov (United States)

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

    2010-04-01

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

  14. POTENCY OF VIBRIO ISOLATES FOR BIOCONTROL OF VIBRIOSIS IN TIGER SHRIMP (PENAEUS MONODON LARVAE

    Directory of Open Access Journals (Sweden)

    B. W. LAY

    2003-01-01

    Full Text Available This study was carried out to obtain Vibrio isolates able to function as biocontr ol of vibriosis in shrimp hatchery. Thirty one Vibrio isolates were isolated from tiger shrimp larvae and hatchery environments, i.e. Labuan, Pangandaran, and Lampung, Indonesia. Pathogenic V. harveyi MR5339 was obtained from Maros, South-Sulawesi and was made as a rifampicin resistant mutant (RFR to screen for those 31 Vibrio isolates in in vitro assays and to allow us to monitor their presence in shrimp larvae and larval rearing water. Almost all Vibrio isolates could inhibit the growth of pathogenic V. harveyi MR5339 RFR. SKT-b isolate from Skeletonema was the most effective to inhibit the growth of V. harveyi MR5339 Rf* and significantly reduced larval mortality in pathogen challenge assays. These prospective biocontrol bacteria, at concentration of 10" CFU/ml, did not show pathogenicity to shrimp larvae. SKT-b was Gram negative, short rod-shape, exhibited yellow colonies on TCBS and swarming on SWC-agar media, motile, u tilized glucose and sucrose but not lactose: produced extra-cellular protease and amylase, but did not produce chitmase. Partial sequencing of 16S-rRNA gene SKT-b showed SKT-b similarity to Vibrio alginofyticus.

  15. Antagonistic Activities of Purple Non-sulfur Bacterial Extracts Against Antibiotic Resistant Vibrio sp.

    Directory of Open Access Journals (Sweden)

    Chandrasekaran, R.

    2011-01-01

    Full Text Available Solvent extracts of native purple non-sulfur bacterial (PNSB isolates from the effluents of brackish shrimp culture ponds, near Nagapattinam coast (South India were evaluated for antibacterial activity by the disc diffusion method. Best results were shown by the chloroform extracts against oxytetracycline resistant Vibrio harveyi and Vibrio fischerii. Among the purple non-sulfur bacterial isolates, Rhodobacter sphaeroides, showed maximum antagonistic activity. The findings suggest that the antagonistic extracts from Rba. sphaeroides could be used as an effective antibiotic in controlling Vibrio spp., in aquaculture systems.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. QStatin, a Selective Inhibitor of Quorum Sensing in Vibrio Species

    Directory of Open Access Journals (Sweden)

    Byoung Sik Kim

    2018-01-01

    Full Text Available Pathogenic Vibrio species cause diseases in diverse marine animals reared in aquaculture. Since their pathogenesis, persistence, and survival in marine environments are regulated by quorum sensing (QS, QS interference has attracted attention as a means to control these bacteria in aquatic settings. A few QS inhibitors of Vibrio species have been reported, but detailed molecular mechanisms are lacking. Here, we identified a novel, potent, and selective Vibrio QS inhibitor, named QStatin [1-(5-bromothiophene-2-sulfonyl-1H-pyrazole], which affects Vibrio harveyi LuxR homologues, the well-conserved master transcriptional regulators for QS in Vibrio species. Crystallographic and biochemical analyses showed that QStatin binds tightly to a putative ligand-binding pocket in SmcR, the LuxR homologue in V. vulnificus, and changes the flexibility of the protein, thereby altering its transcription regulatory activity. Transcriptome analysis revealed that QStatin results in SmcR dysfunction, affecting the expression of SmcR regulon required for virulence, motility/chemotaxis, and biofilm dynamics. Notably, QStatin attenuated representative QS-regulated phenotypes in various Vibrio species, including virulence against the brine shrimp (Artemia franciscana. Together, these results provide molecular insights into the mechanism of action of an effective, sustainable QS inhibitor that is less susceptible to resistance than other antimicrobial agents and useful in controlling the virulence of Vibrio species in aquacultures.

  18. Genomic diversity of vibrios associated with the Brazilian coral Mussismilia hispida and its sympatric zoanthids (Palythoa caribaeorum, Palythoa variabilis and Zoanthus solanderi).

    Science.gov (United States)

    Chimetto, L A; Brocchi, M; Gondo, M; Thompson, C C; Gomez-Gil, B; Thompson, F L

    2009-06-01

    A taxonomic survey of the vibrios associated with the Brazilian endemic coral Mussismilia hispida and the sympatric zoanthids (i.e. Palythoa caribaeorum, Palythoa variabilis and Zoanthus solanderi). Mucus of 54 cnidarian specimens collected in three different places at São Sebastião in two consecutive years (i.e. 2005 and 2006) was used for taxonomic characterization of the cnidarian microbiota. Ninety-eight of the 151 vibrio isolates fell within the vibrio core group according to partial 16S rDNA sequences. We performed the sequencing of recA and pyrH genes of all vibrio isolates. The most abundant taxa belonged to the vibrio core group (Vibrio harveyi, Vibrio rotiferianus, Vibrio campbellii and Vibrio alginolyticus), Vibrio mediterranei (=Vibrio shillonii) and Vibrio chagasii. With the exception of V. chagasii which was found only in the mucus of M. hispida, the other species appeared in different hosts with no evidence for the presence of host-specific clones or species. Using rep-PCR analysis, we observed a high genomic heterogeneity within the vibrios. Each vibrio isolate generated a different rep-PCR fingerprint pattern. There was a complete agreement between the grouping based on rep-PCR and concatenated sequences of pyrH, recA and 16S rDNA, but the pyrH gene has the highest discriminatory power for vibrio species identification. The vibrio core group is dominant in the mucus of these cnidarians. There is a tremendous diversity of vibrio lineages within the coral mucus. pyrH gene sequences permit a clear-cut identification of vibrios. The taxonomic resolution provided by pyrH (but not recA) appears to be enough for identifying species of vibrios and for disclosing putative new taxa. The vibrio core group appears to be dominant in the mucus of the Brazilian cnidarians. The overrepresentation of these vibrios may reflect as yet unknown ecological functions in the coral holobiont.

  19. QStatin, a Selective Inhibitor of Quorum Sensing in Vibrio Species.

    Science.gov (United States)

    Kim, Byoung Sik; Jang, Song Yee; Bang, Ye-Ji; Hwang, Jungwon; Koo, Youngwon; Jang, Kyung Ku; Lim, Dongyeol; Kim, Myung Hee; Choi, Sang Ho

    2018-01-30

    Pathogenic Vibrio species cause diseases in diverse marine animals reared in aquaculture. Since their pathogenesis, persistence, and survival in marine environments are regulated by quorum sensing (QS), QS interference has attracted attention as a means to control these bacteria in aquatic settings. A few QS inhibitors of Vibrio species have been reported, but detailed molecular mechanisms are lacking. Here, we identified a novel, potent, and selective Vibrio QS inhibitor, named QStatin [1-(5-bromothiophene-2-sulfonyl)-1H-pyrazole], which affects Vibrio harveyi LuxR homologues, the well-conserved master transcriptional regulators for QS in Vibrio species. Crystallographic and biochemical analyses showed that QStatin binds tightly to a putative ligand-binding pocket in SmcR, the LuxR homologue in V. vulnificus , and changes the flexibility of the protein, thereby altering its transcription regulatory activity. Transcriptome analysis revealed that QStatin results in SmcR dysfunction, affecting the expression of SmcR regulon required for virulence, motility/chemotaxis, and biofilm dynamics. Notably, QStatin attenuated representative QS-regulated phenotypes in various Vibrio species, including virulence against the brine shrimp ( Artemia franciscana ). Together, these results provide molecular insights into the mechanism of action of an effective, sustainable QS inhibitor that is less susceptible to resistance than other antimicrobial agents and useful in controlling the virulence of Vibrio species in aquacultures. IMPORTANCE Yields of aquaculture, such as penaeid shrimp hatcheries, are greatly affected by vibriosis, a disease caused by pathogenic Vibrio infections. Since bacterial cell-to-cell communication, known as quorum sensing (QS), regulates pathogenesis of Vibrio species in marine environments, QS inhibitors have attracted attention as alternatives to conventional antibiotics in aquatic settings. Here, we used target-based high-throughput screening to identify

  20. Vibrio rotiferianus sp. nov., isolated from cultures of the rotifer Brachionus plicatilis.

    Science.gov (United States)

    Gomez-Gil, B; Thompson, F L; Thompson, C C; Swings, J

    2003-01-01

    Five Gram-negative bacterial strains, oxidase-positive, motile by means of more than one polar flagella, facultative anaerobe, arginine dihydrolase-negative, lysine- and omithine decarboxylase-positive, sensitive to the vibriostatic agent O/129, were isolated from a flow-through rotifer culture system in Gent, Belgium, and previously characterized by fluorescent amplified fragment length polymorphism. Comparison of the 16S rDNA sequence of strain LMG 21460T indicated close relationships (approximately 99% similarity) to Vibrio campbellii, Vibrio harveyi, Vibrio alginolyticus and Vibrio parahaemolyticus. However, DNA hybridization experiments revealed similarity values below 70% with its closest species V. campbellii and V. harveyi. Additionally, the analysed strains differ from related Vibrio species by the utilization of melibiose and production of acid from L-arabinose and amygdalin. Among the strains analysed, differences were observed in some phenotypic characters, particularly susceptibility to ampicillin, polymyxin B and amikacin, and urease activity. The major fatty acids identified were 16:0, 18:1 omega7c, 14:0, 12:0 3-OH and 18:0. Vibrio rotiferianus sp. nov. is proposed, with type strain LMG 21460T (=CAIM 577T); it has a DNA G+C content of 44.5 +/- 0.01 mol%.

  1. Zernike phase contrast cryo-electron tomography of sodium-driven flagellar hook-basal bodies from Vibrio alginolyticus.

    Science.gov (United States)

    Hosogi, Naoki; Shigematsu, Hideki; Terashima, Hiroyuki; Homma, Michio; Nagayama, Kuniaki

    2011-01-01

    Vibrio alginolyticus use flagella to swim. A flagellum consists of a filament, hook and basal body. The basal body is made up of a rod and several ring structures. This study investigates the structure of the T ring which is a unique component of the V. alginolyticus sodium ion-driven flagellar basal body. Using Zernike phase contrast (ZPC) cryo-electron tomography, we compared the 3D structures of purified hook-basal bodies (HBB) from a wild-type strain (KK148) and a deletion mutant lacking MotX and MotY (TH3), which are thought to form the T ring. ZPC images of HBBs had highly improved signal-to-noise ratio compared to conventional phase contrast images. We observed the outline of the HBBs from strains KK148 and TH3, and the TH3 mutant was missing its T ring. In the wild-type strain, the T ring was beneath the LP ring and seemed to form a ring shape with diameter of 32 nm. Copyright © 2010 Elsevier Inc. All rights reserved.

  2. Identification of DLD, by immunoproteomic analysis and evaluation as a potential vaccine antigen against three Vibrio species in Epinephelus coioides.

    Science.gov (United States)

    Pang, Huanying; Chen, Liming; Hoare, Rowena; Huang, Yucong; ZaoheWu; Jian, Jichang

    2016-02-24

    Vibrio spp. represent a serious threat to the culture of Epinephelus coioides (Orange-spotted Grouper) in Southeast Asia. In this study we used two-dimensional electrophoresis (2-DE) and Western blotting to identify common immunogenic proteins of Vibrio alginolyticus, Vibrio harveyi and Vibrio parahaemolyticus. Membranes were probed with orange-spotted grouper anti-V. alginolyticus sera and accordingly 60, 58 and 48 immunogenic protein spots were detected. By matching analysis for the three Western blotting membranes, 6 cross immunogenic spots for the three Vibrio species were identified. They were Outer membrane protein W (OmpW), dihydrolipoamide dehydrogenase (DLD), succinate dehydrogenase flavoprotein subunit(SDHA), elongation factor Ts(Ts), peptide ABC transporter periplasmic peptide-binding protein and phosphoenolpyruvate carboxykinase(PEPCK). One of the proteins, DLD, was used to evaluate the cross protective function for E. coioides with a bacterial immunization and challenge method. The relative percent survival rate of E. coioides against V. alginolyticus, V. harveyi and V. parahaemolyticus was 90%, 86% and 80%, respectively. This work may provide potential cross protective vaccine candidate antigens for three Vibrio species, and DLD may be considered as an effective cross-protective immunogen against three Vibrio species. Copyright © 2016. Published by Elsevier Ltd.

  3. Immune priming and portal of entry effectors improve response to vibrio infection in a resistant population of the European abalone.

    Science.gov (United States)

    Dubief, Bruno; Nunes, Flavia L D; Basuyaux, Olivier; Paillard, Christine

    2017-01-01

    Since 1997, populations of the European abalone Haliotis tuberculata suffer mass mortalities attributed to the bacterium Vibrio harveyi. These mortalities occur at the spawning season, when the abalone immune system is depressed, and when temperatures exceed 17 °C, leading to favorable conditions for V. harveyi proliferation. In order to identify mechanisms of disease resistance, experimental successive infections were carried out on two geographically distinct Brittany populations: one that has suffered recurrent mortalities (Saint-Malo) and one that has not been impacted by the disease (Molène). Furthermore, abalone surviving these two successive bacterial challenges and uninfected abalone were used for several post-infection analyses. The Saint-Malo population was found to be resistant to V. harveyi infection, with a survival rate of 95% compared to 51% for Molène. While in vitro quantification of phagocytosis by flow cytometry showed strong inhibition following the first infection, no inhibition of phagocytosis was observed following the second infection for Saint-Malo, suggesting an immune priming effect. Moreover, assays of phagocytosis of GFP-labelled V. harveyi performed two months post-infection show an inhibition of phagocytosis by extracellular products of V. harveyi for uninfected abalone, while no effect was observed for previously infected abalone from Saint-Malo, suggesting that the effects of immune priming may last upwards of two months. Detection of V. harveyi by qPCR showed that a significantly greater number of abalone from the susceptible population were positive for V. harveyi in the gills, indicating that portal of entry effectors may play a role in resistance to the disease. Collectively, these results suggest a potential synergistic effect of gills and hemolymph in the resistance of H. tuberculata against V. harveyi with an important involvement of the gills, the portal of entry of the bacteria. Copyright © 2016 Elsevier Ltd

  4. Architecture of the Vibrio cholerae toxin-coregulated pilus machine revealed by electron cryotomography

    DEFF Research Database (Denmark)

    Chang, Yi Wei; Kjær, Andreas; Ortega, Davi R.

    2017-01-01

    ,2. T4aP are more widespread and are involved in cell motility 3, DNA transfer 4, host predation 5 and electron transfer 6. T4bP are less prevalent and are mainly found in enteropathogenic bacteria, where they play key roles in host colonization 7. Following similar work on T4aP machines 8,9, here we...... sequence homology to components of the previously analysed Myxococcus xanthus T4aP machine (T4aPM), we find that their structures are nevertheless remarkably similar. Based on homologies with components of the M. xanthus T4aPM and additional reconstructions of TCPM mutants in which the non...

  5. Virulence of luminescent and non-luminescent isogenic vibrios towards gnotobiotic Artemia franciscana larvae and specific pathogen-free Litopenaeus vannamei shrimp.

    Science.gov (United States)

    Phuoc, L H; Defoirdt, T; Sorgeloos, P; Bossier, P

    2009-04-01

    This study was conducted to test the virulence of luminescent (L) and non-luminescent (NL) isogenic strains of Vibrio campbellii LMG21363, Vibrio harveyi BB120 (wild type) and quorum-sensing mutant strains derived from the wild type such as Vibrio harveyi BB152, BB170, MM30 and BB886. The NL strains could be obtained by culturing rifampicin-resistant luminescent strains in the dark under static condition. The virulence of the L and NL strains was tested in gnotobiotic Artemia franciscana larvae challenged with 10(4) CFU ml(-1) of bacteria. All luminescent isogenic tested strains showed higher virulence compared to the NL strains. The virulence of L and NL V. campbellii and V. harveyi BB120 was also tested in specific pathogen-free juvenile shrimp upon intramuscular injection with 10(6) CFU of bacteria. In contrast with Artemia, there was no significant difference in mortality between the groups challenged with L and NL strains (P > 0.05). The non-luminescent strains were not able to revert back to the luminescent state and quorum sensing did not influence this phenotypic shift. Luminescent Vibrio strains can switch to a non-luminescent state by culturing them in static conditions. The NL strains become less virulent as verified in Artemia. The luminescent state of Vibrio cells in a culture needs to be verified in order to assure maintenance of virulence.

  6. Vibrio communis sp. nov., isolated from the marine animals Mussismilia hispida, Phyllogorgia dilatata, Palythoa caribaeorum, Palythoa variabilis and Litopenaeus vannamei.

    Science.gov (United States)

    Chimetto, Luciane A; Cleenwerck, Ilse; Alves, Nelson; Silva, Bruno Sergio; Brocchi, Marcelo; Willems, Anne; De Vos, Paul; Thompson, Fabiano L

    2011-02-01

    Eight Vibrio isolates originating from the marine corals Mussismilia hispida and Phyllogorgia dilatata and the zoanthids Palythoa caribaeorum and Palythoa variabilis in Brazil and the Pacific white shrimp (Litopenaeus vannamei) in Ecuador were studied by means of a polyphasic approach. The novel isolates formed a tight monophyletic group in the genus Vibrio and were closely related to species of the Vibrio harveyi group, to which they showed more than 99 % 16S rRNA gene sequence similarity. Analysis based on concatenated sequences of the following seven genes, 16S rRNA, gyrB, recA, rpoA, topA, pyrH and mreB (5633 bp in length), showed clear separation between the isolates and species of the V. harveyi group. Amplified fragment length polymorphism (AFLP) analysis, performed previously, revealed that a representative isolate of this group, LMG 20370, was clearly separate from known Vibrio species (it belonged to the so-called AFLP cluster A31). DNA-DNA hybridization (DDH) experiments with representative isolates and type strains of the V. harveyi species group revealed high DDH between the novel isolates (more than 74 %) and less than 70 % DDH towards type strains of related Vibrio species, proving the novel species status of the isolates. Phenotypically, the novel species belongs to the arginine dihydrolase (A)-negative, lysine decarboxylase (L)-positive and ornithine decarboxylase (O)-positive (A-/L+/O+) cluster reported previously. Most species of the V. harveyi group (i.e. Vibrio rotiferianus, V. harveyi, V. parahaemolyticus and V. alginolyticus) also belong to this A-/L+/O+ cluster. However, several phenotypic features can be used for the identification of the novel species. In contrast to its closest phylogenetic neighbours, the novel species exhibits esterase (C4) and N-acetyl-β-glucosaminidase activities, but it does not produce acetoin, does not use citrate, α-ketoglutaric acid or propionic acid and does not ferment melibiose. The novel species can

  7. 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.

  8. 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

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

    Directory of Open Access Journals (Sweden)

    Van Calenbergh Serge

    2008-09-01

    Full Text Available Abstract Background To date, only few compounds targeting the AI-2 based quorum sensing (QS system are known. In the present study, we screened cinnamaldehyde and substituted cinnamaldehydes for their ability to interfere with AI-2 based QS. The mechanism of QS inhibition was elucidated by measuring the effect on bioluminescence in several Vibrio harveyi mutants. We also studied in vitro the ability of these compounds to interfere with biofilm formation, stress response and virulence of Vibrio spp. The compounds were also evaluated in an in vivo assay measuring the reduction of Vibrio harveyi virulence towards Artemia shrimp. Results Our results indicate that cinnamaldehyde and several substituted derivatives interfere with AI-2 based QS without inhibiting bacterial growth. The active compounds neither interfered with the bioluminescence system as such, nor with the production of AI-2. Study of the effect in various mutants suggested that the target protein is LuxR. Mobility shift assays revealed a decreased DNA-binding ability of LuxR. The compounds were further shown to (i inhibit biofilm formation in several Vibrio spp., (ii result in a reduced ability to survive starvation and antibiotic treatment, (iii reduce pigment and protease production in Vibrio anguillarum and (iv protect gnotobiotic Artemia shrimp against virulent Vibrio harveyi BB120. Conclusion Cinnamaldehyde and cinnamaldehyde derivatives interfere with AI-2 based QS in various Vibrio spp. by decreasing the DNA-binding ability of LuxR. The use of these compounds resulted in several marked phenotypic changes, including reduced virulence and increased susceptibility to stress. Since inhibitors of AI-2 based quorum sensing are rare, and considering the role of AI-2 in several processes these compounds may be useful leads towards antipathogenic drugs.

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

    Science.gov (United States)

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

    2008-01-01

    Background To date, only few compounds targeting the AI-2 based quorum sensing (QS) system are known. In the present study, we screened cinnamaldehyde and substituted cinnamaldehydes for their ability to interfere with AI-2 based QS. The mechanism of QS inhibition was elucidated by measuring the effect on bioluminescence in several Vibrio harveyi mutants. We also studied in vitro the ability of these compounds to interfere with biofilm formation, stress response and virulence of Vibrio spp. The compounds were also evaluated in an in vivo assay measuring the reduction of Vibrio harveyi virulence towards Artemia shrimp. Results Our results indicate that cinnamaldehyde and several substituted derivatives interfere with AI-2 based QS without inhibiting bacterial growth. The active compounds neither interfered with the bioluminescence system as such, nor with the production of AI-2. Study of the effect in various mutants suggested that the target protein is LuxR. Mobility shift assays revealed a decreased DNA-binding ability of LuxR. The compounds were further shown to (i) inhibit biofilm formation in several Vibrio spp., (ii) result in a reduced ability to survive starvation and antibiotic treatment, (iii) reduce pigment and protease production in Vibrio anguillarum and (iv) protect gnotobiotic Artemia shrimp against virulent Vibrio harveyi BB120. Conclusion Cinnamaldehyde and cinnamaldehyde derivatives interfere with AI-2 based QS in various Vibrio spp. by decreasing the DNA-binding ability of LuxR. The use of these compounds resulted in several marked phenotypic changes, including reduced virulence and increased susceptibility to stress. Since inhibitors of AI-2 based quorum sensing are rare, and considering the role of AI-2 in several processes these compounds may be useful leads towards antipathogenic drugs. PMID:18793453

  11. 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.

  12. Screening and evaluation of local bacteria isolated from shellfish as potential probiotics against pathogenic Vibrios.

    Science.gov (United States)

    Jasmin, M Y; Wagaman, Hazimah; Yin, Tan Ai; Ina-salwany, M Y; Daud, H M; Karim, Murni

    2016-07-01

    The present study was carried out to isolate, screen and evaluate potential candidates of local bacteria isolated from tiger shrimp Penaeus monodon and slipper cupped oysters Crassostrea iredalei as probiotics in shellfish aquaculture. A total of 144 of bacteria were successfully isolated from the intestine and stomach of 20 tails of healthy adult tiger shrimp P. monodon, while 136 were successfully isolated from the digestive tract, gills and inner shells of 10 healthy adult C. iredalei. The number of potential isolates was narrowed down to two from tiger shrimp, and one from slipper cupped oyster after in vitro screening assays. The three isolates, labeled as G11, I24 and S66, were identified as Virgibacillus sp., Bacillus sp. and Exiquobacterium sp., respectively, using 16S rDNA gene analysis. The antagonistic ability of the isolates towards Vibrio alginolyticus and Vibrio harveyi were conducted in stagnant and liquid modes via spot lawn and broth co-culture assay, respectively. In these assays, all the potential probionts were inhibitory to both pathogenic vibrios. In the in-vivo assay, Artemia was used as host and treated with different concentrations of potential probionts (10(4), 10(6) and 10(8) CFU ml(-1)), and challenged with V. alginolyticus and V. harveyi at 105 CFU ml(-1), respectively. Artemia treated with probiont G11 at all concentrations and challenged with V. alginolyticus had increased survival (70 ? 80 %), which was significantly higher as compared with group with only the pathogen (20 %). Meanwhile, probiont I24 increased the survival of Artemia by 70 % at a concentration of 10(8) CFU ml(-1) after being challenged with V. alginolyticus and Artemia treated with 10(6) CFU ml(-1) of probiont S66 had increased survival of 90% after being challenged with V. harveyi. Thus, the three isolates might have potential applications as probiotics in shellfish aquaculture against vibriosis. ?

  13. Reducing Vibrio load in Artemia nauplii using antimicrobial photodynamic therapy: a promising strategy to reduce antibiotic application in shrimp larviculture

    Science.gov (United States)

    Asok, Aparna; Arshad, Esha; Jasmin, C.; Somnath Pai, S.; Bright Singh, I. S.; Mohandas, A.; Anas, Abdulaziz

    2012-01-01

    Summary We propose antimicrobial photodynamic therapy (aPDT) as an alternative strategy to reduce the use of antibiotics in shrimp larviculture systems. The growth of a multiple antibiotic resistant Vibrio harveyi strain was effectively controlled by treating the cells with Rose Bengal and photosensitizing for 30 min using a halogen lamp. This resulted in the death of > 50% of the cells within the first 10 min of exposure and the 50% reduction in the cell wall integrity after 30 min could be attributed to the destruction of outer membrane protein of V. harveyi by reactive oxygen intermediates produced during the photosensitization. Further, mesocosm experiments with V. harveyi and Artemia nauplii demonstrated that in 30 min, the aPDT could kill 78.9% and 91.2% of heterotrophic bacterial and Vibrio population respectively. In conclusion, the study demonstrated that aPDT with its rapid action and as yet unreported resistance development possibilities could be a propitious strategy to reduce the use of antibiotics in shrimp larviculture systems and thereby, avoid their hazardous effects on human health and the ecosystem at large. PMID:21951316

  14. Virulence of luminous vibrios to Artemia franciscana nauplii.

    Science.gov (United States)

    Soto-Rodriguez, S A; Roque, A; Lizarraga-Partida, M L; Guerra-Flores, A L; Gomez-Gill, B

    2003-02-27

    From healthy and diseased penaeid shrimp from Asia and the Americas, 25 luminous and 2 non-luminous bacterial strains were isolated, and 14 were phenotypically identified as Vibrio harveyi; 9 isolates produced significant mortalities (45 to 80%) in Artemia franciscana nauplii at inoculation densities of 10(5) to 10(6) CFU ml(-1) compared to the controls (unchallenged nauplii). The maximum number of bacteria ingested (bioencapsulated) by the Artemia nauplii varied from less than 10 to 10(3) CFU nauplius(-1) and no significant relationship was observed between the density of bacteria inoculated, the amount of bacteria ingested, and naupliar mortality. Significant correlations were obtained between naupliar mortality and production of proteases, phospholipases or siderophores, but not between mortality and lipase production, gelatinase production, hydrophobicity or hemolytic activity. The results suggest that virulence of the strains tested was more related to the production of particular exoenzymes than to the measured colonization factors.

  15. Phytoplankton production systems in a shellfish hatchery: variations of the bacterial load and diversity of vibrios.

    Science.gov (United States)

    Dubert, J; Fernández-Pardo, A; Nóvoa, S; Barja, J L; Prado, S

    2015-06-01

    Outbreaks of disease caused by some Vibrio species represent the main production bottleneck in shellfish hatcheries. Although the phytoplankton used as food is one of the main sources of bacteria, studies of the associated bacterial populations, specifically vibrios, are scarce. The aim of the study was the microbiological monitoring of the microalgae as the first step in assessing the risk disease for bivalve cultures. Two phytoplankton production systems were sampled weekly throughout 1-year period in a bivalve hatchery. Quantitative analysis revealed high levels of marine heterotrophic bacteria in both systems throughout the study. Presumptive vibrios were detected occasionally and at low concentrations. In most of the cases, they belonged to the Splendidus and Harveyi clades. The early detection of vibrios in the microalgae may be the key for a successful bivalve culture. Their abundance and diversity were affected by factors related to the hatchery environment. This work represents the first long study where the presence of vibrios was evaluated rigorously in phytoplankton production systems and provides a suitable microbiological protocol to control and guarantee the quality of the algal cultures to avoid the risk of transferring potential pathogens to shellfish larvae and/or broodstock. © 2015 The Society for Applied Microbiology.

  16. 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.

    , National Institute of Oceanography Regional Centre (CSIR), Kochi- 682 018, India b Department of Microbiology, College of Fisheries, Mangalore-575 002, India c Department of Biology, Eastern New Mexico University, Portales, New Mexico 88130 USA... institutional project SIP 1302 is gratefully acknowledged. S.K is grateful to Prof. (Dr). Manuel Varela, Department of Biology, Eastern New Mexico University, USA for scientific advice and guidance. This is NIO contribution no. 4508.   10...

  17. Illuminating Cell Signaling: Using "Vibrio harveyi" in an Introductory Biology Laboratory

    Science.gov (United States)

    Hrizo, Stacy L.; Kaufmann, Nancy

    2009-01-01

    Cell signaling is an essential cellular process that is performed by all living organisms. Bacteria communicate with each other using a chemical language in a signaling pathway that allows bacteria to evaluate the size of their population, determine when they have reached a critical mass (quorum sensing), and then change their behavior in unison…

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

    Digital Repository Service at National Institute of Oceanography (India)

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

    at the early life-stages of shrimps are of great significance. An understanding therefore, of molecular responses and defense mechanism(s) in shrimps against bacterial pathogens is essential. Besides production of antimicrobial biomolecules, the innate... of this strain grown for 24h in seawater nutrient broth were used for the experimental challenge. Cells were harvested by centrifugation and re-suspended in sterilized seawater and the PL were ‘bath’ challenged by adding the harvested cells to attain 2.5- 3...

  19. Studies on luminous, Vibrio harveyi associated with shrimp culture system rearing Panaeus monodon

    Digital Repository Service at National Institute of Oceanography (India)

    Kannapiran, E.; Ravindran, J.; Chandrasekar, R.; Kalaiarasi, A.

    feed, fertilizers and vitamins. This observation reconciles with the earlier findings that the artificial feed and other organic additions in the aquaculture farms enhanced the heterotrophic bacterial population (MacLean et al., 1994; Sharmila et al... the pond bottom preparation and low level of organic matter in the pond. As culture period progresses, accumulation of organic wastes in the shrimp pond takes placed due to the application of organic manure and fertilizers, feed waste, fecal matter...

  20. Quorum quenching bacteria protect Macrobrachium rosenbergii larvae from Vibrio harveyi infection.

    Science.gov (United States)

    Nhan, D T; Cam, D T V; Wille, M; Defoirdt, T; Bossier, P; Sorgeloos, P

    2010-09-01

    In this study, we investigated the effect of N-acyl homoserine lactone-degrading bacterial enrichment cultures (ECs) on larviculture of the giant freshwater prawn Macrobrachium rosenbergii. The larval performance in terms of larval growth, larval survival, larval quality, duration of the larval rearing process and microflora levels in the rearing water as well as inside the prawn gut was investigated. The application of the EC bacteria was performed in two ways: by adding them directly into the larval rearing water and via enriched Artemia nauplii used for larval feeding. The results of the study demonstrated that both ECs that were tested had a similar positive effect on larval survival and larval quality, whereas they did not affect larval growth or the duration of the larval rearing process. Under normal hatchery conditions, the optimal EC densities were found to be 10(6) CFU ml(-1) for adding into the rearing water and 5 × 10(8) CFU ml(-1) for enrichment of Artemia nauplii used for feeding of the larvae. In the hatchery, the ECs can be grown on waste streams of Artemia hatching. Application of this kind of ECs could lead to a more sustainable aquaculture production, by replacing the use of antibiotics to control diseases. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

  1. Distribution of putative virulence genes and antimicrobial drug resistance in Vibrio harveyi

    Digital Repository Service at National Institute of Oceanography (India)

    Parvathi, A.; Mendez, D.; Anto, C.

    zonula occludens toxin (Zot) and a hemolysin-coregulated protein gene (hcp) by polymerase chain reaction (PCR). Of the four putative reversible toxin genes, vhh-1 was detected in 31% of the isolates, vhh-2 in 46%, vhh-3 in 23% and vhh-4 was detected in 27...

  2. 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 of...

  3. Single nucleotide polymorphisms in i-type lysozyme gene and their correlation with vibrio-resistance and growth of clam Meretrix meretrix based on the selected resistance stocks.

    Science.gov (United States)

    Yue, Xin; Wang, Hongxia; Huang, Xiaohong; Wang, Chao; Chai, Xueliang; Wang, Chunde; Liu, Baozhong

    2012-09-01

    I-type lysozyme is considered to play crucial roles in both anti-bacteria and digestion function of the bivalve, which signifies that it is related to both immunity and growth. In this study, based on the principle of case-control association analysis, using the stock materials with different vibrio-resistance profile obtained by selective breeding, single nucleotide polymorphisms (SNPs) in the DNA partial sequence of an i-type lysozyme of Meretrix meretrix (MmeLys) were discovered and examined for their association with vibrio-resistance and growth. Twenty-seven SNPs were detected and fifteen of them were genotyped in clam stocks with different resistance to Vibrio harveyi (09-C and 09-R) and to Vibrio parahaemolyticus (11-S and 11-R). Allele frequency distribution among different stocks was compared. And wet weight of clams with different genotype at each SNP locus was compared. The results indicated that SNP locus 9 was associated with V. harveyi and V. parahaemolyticus resistance and growth of M. meretrix. Loci 12 and 14 were associated with both V. parahaemolyticus-resistance and growth, and also have the potential to be related with V. harveyi-resistance of M. meretrix. Therefore these three SNPs especially locus 9 were the potential markers which may be involved in assisting resistance selective breeding. In addition, this study showed evidence that improvements in clam resistance to vibriosis could be achieved through selective breeding. All results provided encouragement for the continuation of the selective breeding program for vibrio-resistance gain in clam M. meretrix and the application of polymorphisms in MmeLys to the future marker assisted selection. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. 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

    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...... 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......-producing Phaeobacter isolated from Mediterranean marine larviculture are promising probiotic bacteria against pathogenic Vibrio in crustacean live-feed cultures for marine fish larvae....

  5. Molecular Cloning and Expression Analysis of Transforming Growth Factor TGF-β1 and TGF-β3 in Half-smooth Tongue Sole (Cynoglossus semilaevis) Following Stimulation with Vibrio harveyi%半滑舌鳎转化生长因子TGF-β1和TGF-β3基因的克隆及受哈维氏弧菌感染后表达分析

    Institute of Scientific and Technical Information of China (English)

    李雪; 陈松林; 杨长庚; 邵长伟; 李仰真; 位战飞

    2016-01-01

    转化生长因子β (transforming growth factor β,TGF-β)是一类具有多种功能的蛋白超家族,在细胞免疫、细胞增殖分化和组织损伤的修复中起着关键性作用.本研究从半滑舌鳎(Cynoglossus semilaevis)肝脏中克隆获得了TGF-β1和TGF-β3基因.推导的TGF-β1和TGF-β3氨基酸序列均含有多个N糖基化位点和一个TGF-β家族标签.系统进化树分析显示,半滑舌鳎TG F-β1和TGF-β3分别与鱼类的TGF-β1和TGF-β3亲缘关系最为密切.qRT-PCR结果表明,半滑舌鳎TGF-β1和TGF-β3基因在健康鱼的多个组织中均有表达,二者在皮肤中表达量最高,在肌肉中表达量最低.经哈维氏弧菌(Vibrio harveyi)感染后,TGF-β1在肝脏、脾脏和肾脏中呈现先上升后下降的表达趋势,在感染48 h后的肝脏中表达量达到最大值,是对照组的3.17倍;TGF-β3在脾脏、肾脏和鳃中也呈现先上升后下降的表达趋势,在感染24 h后的鳃中表达量达到最大值,是对照组的4.71倍.以上结果提示,TG F-β1和TGF-β3可能在半滑舌鳎抵御细菌感染的免疫中发挥了重要作用,本研究为证明二者参与机体免疫调节提供了有力证据,为半滑舌鳎分子免疫研究提供了理论依据.

  6. The first temporal and spatial assessment of Vibrio diversity of the surrounding seawater of coral reefs in Ishigaki, Japan

    Directory of Open Access Journals (Sweden)

    AKM Rohul Amin

    2016-08-01

    Full Text Available Coral reefs perform a major role in regulating marine biodiversity and serve as hotspot for highly dynamic and diverse microbiomes as holobionts. Corals around Ishigaki, however, are at risk due to tremendous stressors including elevation of seawater temperature, eutrophication and so on. However, no information is currently available on how Vibrio diversity fluctuates spatially and temporally due to environmental determinants in Ishigaki coral reef ecosystems. The aim of this study is to elucidate spatiotemporal Vibrio diversity dynamic at both community and population levels and to assess the environmental drivers correlated to Vibrio abundance and diversity. The Vibrio community identified based on pyrH gene phylogeny of 685 isolates from seawater directly connecting to Ishigaki coral holobionts consisted of 22 known and 12 potential novel Vibrionaceae species. The most prominent species were V. hyugaensis, V. owensii and V. harveyi followed by V. maritimus/V. variabillis, V. campbellii, V. coralliilyticus and P. rosenbergii. The Vibrio community fluctuations, assessed by PCoA with UniFrac distance and clustering with Euclidiean distance were varied less not only by year but also by site. Interestingly, significant positive correlation was observed between rising seawater temperature and the abundance of V. campbellii (r=0.62; P<0.05 whereas the opposite was observed for V. owensii (r=-0.58; P<0.05 and the C6 group of V. hyugaensis (r=-0.62; P<0.05. AdaptML-based microhabitat differentiation revealed that V. harveyi, V. campbellii, P. rosenbergii and V. coralliilyticus populations were less-ecologically distinctive whereas V. astriarenae and V. ishigakensis were ecologically diverse. This knowledge could be important clue for the future actions coral conservation.

  7. Efeito do uso de bactérias probióticas na sobrevivência de larvas de Litopenaeus vannamei expostas à infecção experimental por Vibrio spp.

    OpenAIRE

    Joana Angélica Lyra Vogeley de Carvalho

    2011-01-01

    O presente estudo teve como objetivo avaliar a sobrevivência de larvas de Litopenaeus vannamei tratadas com bactérias probióticas e posteriormente infectadas por Vibrio harveyi e Vibrio alginolyticus. Os camarões foram mantidos em dois tanques de 80L nas mesmas condições, mas apenas um deles foi previamente tratado com probiótico comercial (Bacillus spp.) adicionado diariamente na água na concentração de 105 UFC/ml. Este protocolo foi mantido até os animais alcançarem o estágio de desen...

  8. 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.

  9. 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.

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

    Institute of Scientific and Technical Information of China (English)

    Prabhakaran Priyaja; Puthumana Jayesh; Neil Scolastin Correya; Balachandran Sreelakshmi; Naduthalmuriparambil S Sudheer; Rosamma Philip; Isaac Sarogeni Bright Singh

    2014-01-01

    Objective: To abrogate pathogenic vibrios in aquaculture by testing the potential of Pseudomonas isolates from fresh water, brackish and marine environments as probiotics.Methods: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 werePseudomonas 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:Purification and structural elucidation of antagonistic compound were carried out. ideal for application in freshwater, MCCB102 and MCCB103 in brackish water and MCCB117 and The present investigation showed that Pseudomonas aeruginosa MCCB119 would be MCCB118 in marine aquaculture systems as putative probiotics in the management of vibrios.

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Three vibrio-resistance related EST-SSR markers revealed by selective genotyping in the clam Meretrix meretrix.

    Science.gov (United States)

    Nie, Qing; Yue, Xin; Chai, Xueliang; Wang, Hongxia; Liu, Baozhong

    2013-08-01

    The clam Meretrix meretrix is an important commercial bivalve distributed in the coastal areas of South and Southeast Asia. In this study, marker-trait association analyses were performed based on the stock materials of M. meretrix with different vibrio-resistance profile obtained by selective breeding. Forty-eight EST-SSR markers were screened and 27 polymorphic SSRs of them were genotyped in the clam stocks with different resistance to Vibrio parahaemolyticus (11-R and 11-S) and to Vibrio harveyi (09-R and 09-C). Allele frequency distributions of the SSRs among different stocks were compared using Pearson's Chi-square test, and three functional EST-SSR markers (MM959, MM4765 and MM8364) were found to be associated with vibrio-resistance trait. The 140-bp allele of MM959 and 128-bp allele of MM4765 had significantly higher frequencies in resistant groups (11-R and 09-R) than in susceptive/control groups (11-S and 09-C) (P SSR markers were consistent with the three subgroups distinctions. The putative functions of contig959, contig4765 and contig8364 also suggested that the three SSR-involved genes might play important roles in immunity of M. meretrix. All these results supported that EST-SSR markers MM959, MM4765 and MM8364 were associated with vibrio-resistance and would be useful for marker-assisted selection (MAS) in M. meretrix genetic breeding. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Effects of different doses of skt-b vibrio probiotic bacteria addition on survival and growth rate of tiger shrimp (Penaeus monodon larva

    Directory of Open Access Journals (Sweden)

    . Widanarni

    2010-01-01

    Full Text Available Probiotic bacteria has been widely used as biocontrol agents in tiger shrimp hatcheries.  Vibrio SKT-b is one of the probiotic bacteria candidates that could suppressed the growth of pathogenic bacteria Vibrio harveyi and could increase survival rate of tiger shrimp larva. This experiment was carried out to study the effects of probiotic bacteria SKT-b Vibrio addition at different doses on survival and growth rate of tiger shrimp larva.  Experiment was conducted with five treatments and three replications, consisted of SKT-b Vibrio probiotic bacteria addition at the doses of 103 CFU/ml, 104 CFU/ml, 105 CFU/ml, and 106 CFU/ml and control (0 CFU/ml.  Results showed that optimum dose of probiotic bacteria for tiger shrimp was 104 CFU/ml with a survival rate of 94.67%. However, the addition of probiotic bacteria at this particular dose did not significantly increase shrimp growth rate as compared with control. Key words: Probiotic bacteria, SKT-b Vibrio, doses, tiger shrimp larva   ABSTRAK Bakteri probiotik telah banyak digunakan sebagai agen biokontrol dalam pembenihan udang windu.  Vibrio SKT-b merupakan salah satu jenis bakteri kandidat probiotik yang telah diuji dapat menekan pertumbuhan bakteri patogen Vibrio harveyi dan dapat meningkatkan kelangsungan hidup larva udang windu.  Tujuan dari penelitian ini adalah untuk mengetahui pengaruh pemberian bakteri probiotik Vibrio SKT-b dengan dosis yang berbeda terhadap kelangsungan hidup dan pertumbuhan larva udang windu. Penelitian ini dilakukan dalam 5 perlakuan dengan masing-masing 3 ulangan, yaitu penambahan bakteri probiotik Vibrio SKT-b dengan dosis 103 CFU/ml, 104 CFU/ml, 105 CFU/ml, dan 106 CFU/ml dan kontrol (0 CFU/ml.  Hasil penelitian menunjukkan bahwa dosis optimal untuk larva udang windu adalah 104 CFU/ml dengan nilai kelangsungan hidup 94,67%. Namun, pemberian bakteri probiotik tersebut belum menghasilkan pertumbuhan yang berbeda nyata dengan kontrol. Kata kunci: Bakteri probiotik

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

    Science.gov (United States)

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

    2011-05-01

    Vibrio parahaemolyticus is a gram-negative, halophilic bacterium indigenous to marine and estuarine environments and it is capable of causing food and water-borne illness in humans. It can also cause disease in marine animals, including cultured species. Currently, culture-based techniques are used for quantification of V. parahaemolyticus in environmental samples; however, these can be misleading as they fail to detect V. parahaemolyticus in a viable but nonculturable (VBNC) state which leads to an underestimation of the population density. In this study, we used a novel fluorescence visualization technique, called recognition of individual gene fluorescence in situ hybridization (RING-FISH), which targets chromosomal DNA for enumeration. A polynucleotide probe labeled with Cyanine 3 (Cy3) was created corresponding to the ubiquitous V. parahaemolyticus gene that codes for thermolabile hemolysin (tlh). When coupled with the Kogure method to distinguish viable from dead cells, RING-FISH probes reliably enumerated total, viable V. parahaemolyticus. The probe was tested for sensitivity and specificity against a pure culture of tlh(+), tdh(-), trh(-)V. parahaemolyticus, pure cultures of Vibrio vulnificus, Vibrio harveyi, Vibrio alginolyticus and Vibrio fischeri, and a mixed environmental sample. This research will provide additional tools for a better understanding of the risk these environmental organisms pose to human health. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. The Diversity of Vibrios Associated with Vibriosis in Pacific White Shrimp (Litopenaeus vannamei) from Extensive Shrimp Pond in Kendal District, Indonesia

    Science.gov (United States)

    Sarjito; Harjuno Condro Haditomo, Alfabetian; Desrina; Djunaedi, Ali; Budi Prayitno, Slamet

    2018-02-01

    Vibriosis out breaks frequently occur in extensive shrimps farming. The study were commenced to find out the clinical signs of white shrimp that was infected by the Vibrio and to identify the bacterial associated with vibriosis in the pacific white shrimp, Litopenaeus vannamei. Bacterial isolates were gained from hepatopancreas and telson of moribund shrimps that were collected from extensive shrimp ponds of Kendal District, Indonesia and cultured on Thiosulfate Citrate Bile Salts Sucrose Agar (TCBSA). Isolates were clustered and identified using repetitive sequence-based polymerase chain reaction (rep-PCR). Three representative isolates (SJV 03, SJV 05 and SJV 19) were amplified with PCR using primers for 16S rRNA, and sequence for further identification. The clinical signs of shrimps affected by vibrio were pale hepatopancreas, weak of telson, dark and reddish coloration of smouth, patches of red colour in part of the body on the carapace, periopods, pleuopods, and telson. A total of 19 isolates were obtained and belong to three groups of genus Vibrios. Result of the 16S DNA sequence analysis, the vibrio found in this study related to vibriosis in white shrimps from extensive shrimp ponds of Kendal were closely related to Vibrio harveyi (SJV 03); V. parahaemolyticus (SJV 05) and V. alginolyticus (SJV 19).

  16. RpoS induces expression of the Vibrio anguillarum quorum-sensing regulator VanT.

    Science.gov (United States)

    Weber, Barbara; Croxatto, Antony; Chen, Chang; Milton, Debra L

    2008-03-01

    In vibrios, regulation of the Vibrio harveyi-like LuxR transcriptional activators occurs post-transcriptionally via small regulatory RNAs (sRNAs) that destabilize the luxR mRNA at a low cell population, eliminating expression of LuxR. Expression of the sRNAs is modulated by the vibrio quorum-sensing phosphorelay systems. However, vanT mRNA, which encodes a LuxR homologue in Vibrio anguillarum, is abundant at low and high cell density, indicating that VanT expression may be regulated via additional mechanisms. In this study, Western analyses showed that VanT was expressed throughout growth with a peak of expression during late exponential growth. VanO induced partial destabilization of vanT mRNA via activation of at least one Qrr sRNA. Interestingly, the sigma factor RpoS significantly stabilized vanT mRNA and induced VanT expression during late exponential growth. This induction was in part due to RpoS repressing expression of Hfq, an RNA chaperone. RpoS is not part of the quorum-sensing regulatory cascade since RpoS did not regulate expression or activity of VanO, and RpoS was not regulated by VanO or VanT. VanT and RpoS were needed for survival following UV irradiation and for pigment and metalloprotease production, suggesting that RpoS works with the quorum-sensing systems to modulate expression of VanT, which regulates survival and stress responses.

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

    Directory of Open Access Journals (Sweden)

    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.

  18. Vibrios and Aeromonas.

    Science.gov (United States)

    Holmberg, S D

    1988-09-01

    There are many similarities in the Vibrionaceae that cause human illness in the United States (see Table 1). Vibrios are characteristically indigenous to marine, estuarine, and brackish environments. They are distributed mainly in Gulf of Mexico coastal water, and these organisms "bloom" when the water is warm. Outbreaks of disease in humans frequently occur in summer, coinciding with multiplication of vibrios in warm water. Sporadic cases and small outbreaks of cholera continue to occur in persons living on or near the Gulf of Mexico, but infection in most persons is unrecognized. In fact, more serious and frequent illnesses result from V. vulnificus wound infections and from gastroenteritis caused by vibrios other than V. cholerae 01. Underlying hepatic or neoplastic disease (especially leukemia) apparently increases the likelihood and severity of illnesses caused by V. vulnificus and Aeromonas. Some Vibrionaceae produce clinical illness by means of enterotoxins identical or similar to cholera toxin. For many others, hemolysins, cytotoxins, and other exotoxins are necessary to produce disease; the importance of these virulence factors often is not known or the importance of these virulence factors often is not known or is of doubtful significance. Also, purported pathogenicity as demonstrated by animal models, such as fluid accumulation in ligated ileal loops, is quite nonspecific and needs to be interpreted cautiously. For Plesiomonas, a mode of pathogenesis has not been discovered. Eating raw shellfish (frequently raw oysters) has been linked epidemiologically to enteric infections with most of these bacteria; foreign travel and exposure to seawater are other frequently observed epidemiologic associations with infection. Foreign travel, particularly to the Yucatan Peninsula of Mexico, has been strongly associated with the acquisition of non-01 V. cholerae and Plesiomonas organisms. Most Vibrionaceae in the United States are susceptible in vitro--and illnesses

  19. PREPARATION AND APPLICATIONS OF MONOCLONAL ANTIBODY AGAINST THE PATHOGENIC HARVEYI FROM PSEUDOSCIAENA CROCEA%大黄鱼病原哈维氏弧菌单克隆抗体的制备及其应用

    Institute of Scientific and Technical Information of China (English)

    郝贵杰; 沈锦玉; 徐洋; 姚嘉赟; 潘晓艺; 尹文林

    2009-01-01

    用甲醛灭活哈维氏弧菌(Vibrio harveyi)GYC1108-1制备免疫原免疫8周龄雌性BALB/c小鼠,利用淋巴细胞杂交瘤技术,用间接酶联免疫吸附试验(ELISA)对杂交瘤进行筛选,阳性克隆经2-3次亚克隆后,共获得5株针对GYC1108-1的单克隆抗体.选取1B7制备小鼠腹水抗体,其细胞上清及腹水效价分别为1∶3200和1∶32000.同样用灭活的哈维氏弧菌免疫新西兰大白兔,5次免疫后颈动脉采血,离心取血清,并用饱和硫酸铵法进行纯化,制备了兔抗哈维氏弧菌多克隆抗体,其ELISA效价为1∶51200.利用1B7单克隆抗体和兔抗哈维氏弧菌多克隆抗体以及山羊抗小鼠HRP酶标二抗,建立了检测哈维氏弧菌的三抗体夹心酶联免疫吸附试验(TAS-ELISA)方法.该方法对哈维氏弧菌的最小检出浓度为1×104个/mL.用该TAS-ELISA方法检测大黄鱼(Pseudosciaena crocea)样品,54尾患病大黄鱼中有45尾检出哈维氏弧菌,而14尾健康大黄鱼都没有检出哈维氏弧菌.由此可见,本试验建立的TAS-ELISA方法,可以用于患病大黄鱼哈维氏弧菌的快速诊断.%Vibrio harveyi is a kind of important pathogenic bacteria for seawater animals, such as prawn, crab, calm and ormer. Along with the development of seawater fish cage-culture in China, Vibrio harveyi also becomes a main cause of disease of cage-culture fish, including Lutianus erythopterus, Seriola dumerili , Epinephelus coioides and Pseudosciaena cro-cea. One pathogenic V. harveyi strain, GYC1108-1, which was isolated from diseased Pseudosciaena crocea in Zhejiang Province, 2003, was identified after morphological observation, biochemical characteristic analysis, 16sRNA and HSP60 gene sequence detection.

  20. 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...

  1. Bovine Lactoferrin and Lactoferrin-Derived Peptides Inhibit the Growth of Vibrio cholerae and Other Vibrio species

    Directory of Open Access Journals (Sweden)

    Erika Acosta-Smith

    2018-01-01

    Full Text Available 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.

  2. 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

  3. Fluorescence detection of the pathogenic bacteria Vibrio harveyi in solution and animal cells using semiconductor quantum dots

    Digital Repository Service at National Institute of Oceanography (India)

    Arshad, E.; Anas, A.; Aparna, A.; Jasmin, C.; Pai, S.S.; BrightSingh, I.S.; Mohandas, A.; Biju, V.

    ,1 A. Mohandas,1 Vasudevanpillai Biju4,* 1National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Kochi 682 016, India 2Council of Scientific and Industrial Research (CSIR)- National Institute of Oceanography (NIO...), Regional Centre Cochin, Kochi 682 018, India 3Amity Institute of Virology and Immunology, Amity University, Noida, Uttar Pradesh 201 313 India 4 Health Research Institute, AIST, 2217-14 Hayashi-Cho, Takamatsu, Kagawa 761-0395, Japan *Address...

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

    Digital Repository Service at National Institute of Oceanography (India)

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

    controlled tumor protein (TCTP), hemocyanin, serine carboxy peptidase, and chitinase accounted for 10% of the identified ESTs. Interestingly, there was also a higher incidence of genes related to cell cycle progression and protein modification such as S...). It was earlier reported to be upregulated following challenge with WSSV (Pan et al., 2005). Binding of free iron is considered bacteriostatic, as bacterial cell growth, in particular of Vibrionaceae, is inhibited if iron is not available for synthesis...

  5. Microbial Ecophysiology of Vibrio ruber

    Directory of Open Access Journals (Sweden)

    Tjaša Danevčič

    2014-01-01

    Full Text Available Bacteria use different adaptation strategies to survive environmental perturbations. In this minireview, adaptation strategies of new red-pigmented Vibrio ruber isolated from coastal environments to different environmental stresses (i.e. salinity, viscosity, UV light, mitomycin C, nutrient availability and temperature are reviewed. To cope with environmental stresses Vibrio ruber uses several different adaptive strategies. For example, lipid composition as well as phase behaviour are strongly dependent on salt concentration. Vibrio ruber membrane has no hydroxy fatty acids, but exceptionally high lysolipid content compared to other related Vibrio species. Inorganic nutrient uptake by bacteria is selective, depends on environmental conditions and varies several fold with environmental perturbations. Protein composition, carbon flow through the central metabolic pathways, energy generation as well as secondary metabolite production adapt readily to stress conditions. The activity of glucose-6-phosphate dehydrogenase proved to be a good indicator of Vibrio ruber stress. Cells are able to modulate their local viscosity in response to variations of environmental viscosity. The bacterium harbours several viral genetic elements in its genome, which could be induced by mitomycin C. Environmental conditions during growth of bacteria have a significant effect on lysate carbon turnover. Secondary metabolite prodigiosin confers protection against UV in the environment, which adds to the known repertoire of prodigiosin ecophysiological functions. In conclusion, Vibrio ruber in its short acquaintance with the scientific community (less than ten years has proven to be an immensely valuable model system for ecophysiological studies of bacteria.

  6. Effect of molases addition on survival and growth of tiger shrimp Penaeus monodon Fab. larva treated with SKT-b Vibrio probiotic bacteria

    Directory of Open Access Journals (Sweden)

    , Widanarni

    2015-03-01

    Full Text Available  ABSTRACTProbiotic bacteria SKT-b Vibrio that belong to heterotrophic bacteria have been tested can suppress the growth of pathogenic Vibrio harveyi in tiger shrimp larvae. One of the most important energy sources of heterotrophic bacteria is organic carbon such as sucrose and glucose found in molasses. The objective of this experiment was to study the effect of molasses addition on the survival and growth rate of tiger shrimp Penaeus monodon larvae treated with SKT-b Vibrio probiotic bacteria. This experiment was done both in vitro and in vivo. In vitro tests performed by growing bacteria in media containing molasses, fish meal and premix. Whereas in vivo tests performed with the addition of molasses to shrimp larva culture media inoculated with SKT-b Vibrio bacteria which included five treatments: the control treatment without bacteria and molasses addition (K, the addition of bacteria without molasses (B, the addition of molasses 1 ppm and bacteria (MB1, the addition of molasses 3 ppm and bacteria (MB3, and the addition of molasses 5 ppm and bacteria (MB5. The result of in vitro tests showed the SKT-b Vibrio bacteria can grow on molasses media with a population reached 0,41 × 108 CFU/ml. In vivo test results showed that treatment MB3 gave the highest survival (93.3% and growth weight (35.94%, but were not significantly different from control result in the survival and growth weight that respectively were 83.3% and 30.38%.Keywords: probiotic bacteria, SKT-b Vibrio, molasses, tiger shrimp larvae ABSTRAKBakteri probiotik Vibrio SKT-b yang tergolong bakteri heterotrof telah diuji dapat menekan pertumbuhan Vibrio harveyi yang bersifat patogen pada larva udang windu. Salah satu sumber energi yang paling penting bagi bakteri heterotrof adalah karbon organik seperti sukrosa dan glukosa yang terdapat pada molase. Tujuan dari penelitian ini adalah untuk mengetahui pengaruh penambahan molase terhadap kelangsungan hidup dan pertumbuhan larva udang

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

    Science.gov (United States)

    This study shows that naturally occurring Vibrio predatory bacteria (VPB) exert a major role in controlling pathogenic vibrios in seawater and shellfish. The growth and persistence of Vibrio parahaemolyticus (Vp) and Vibrio vulnificus (Vv) were assessed in natural seawater and in the Eastern oyster...

  8. Vibrio campbellii hmgA-mediated pyomelanization impairs quorum sensing, virulence and cellular fitness

    Directory of Open Access Journals (Sweden)

    Zheng eWang

    2013-12-01

    Full Text Available Melanization due to the inactivation of the homogentisate-1,2-dioxygenase gene (hmgA has been demonstrated to increase stress resistance, persistence and virulence in some bacterial species but such pigmented mutants have not been observed in pathogenic members of the Vibrio Harveyi clade. In this study, we used Vibrio campbellii ATCC BAA-1116 as model organism to understand how melanization affected cellular phenotype, metabolism and virulence. An in-frame deletion of the hmgA gene resulted in the overproduction of a pigment in cell culture supernatants and cellular membranes that was identified as pyomelanin. Unlike previous demonstrations in Vibrio cholerae, Burkholderia cepacia and Pseudomonas aeruginosa, the pigmented V. campbellii mutant did not show increased UV resistance and was found to be ~2.7 times less virulent than the wild type strain in Penaeus monodon shrimp virulence assays. However, the extracted pyomelanin pigment did confer a higher resistance to oxidative stress when incubated with wild type cells. Microarray-based transcriptomic analyses revealed that the hmgA gene deletion and subsequent pyomelanin production negatively effected the expression of 129 genes primarily involved in energy production, amino acid and lipid metabolism, and protein translation and turnover. This transcriptional response was mediated in part by an impairment of the quorum sensing regulon as transcripts of the quorum sensing high cell density master regulator LuxR and other operonic members of this regulon were significantly repressed in the hmgA mutant. Taken together, the results suggest that the pyomelanization of V. campbellii sufficiently impairs the metabolic activities of this organism and renders it less fit and virulent than its isogenic wild type strain.

  9. Vibrio campbellii hmgA-mediated pyomelanization impairs quorum sensing, virulence, and cellular fitness.

    Science.gov (United States)

    Wang, Zheng; Lin, Baochuan; Mostaghim, Anahita; Rubin, Robert A; Glaser, Evan R; Mittraparp-Arthorn, Pimonsri; Thompson, Janelle R; Vuddhakul, Varaporn; Vora, Gary J

    2013-01-01

    Melanization due to the inactivation of the homogentisate-1,2-dioxygenase gene (hmgA) has been demonstrated to increase stress resistance, persistence, and virulence in some bacterial species but such pigmented mutants have not been observed in pathogenic members of the Vibrio Harveyi clade. In this study, we used Vibrio campbellii ATCC BAA-1116 as model organism to understand how melanization affected cellular phenotype, metabolism, and virulence. An in-frame deletion of the hmgA gene resulted in the overproduction of a pigment in cell culture supernatants and cellular membranes that was identified as pyomelanin. Unlike previous demonstrations in Vibrio cholerae, Burkholderia cepacia, and Pseudomonas aeruginosa, the pigmented V. campbellii mutant did not show increased UV resistance and was found to be ~2.7 times less virulent than the wild type strain in Penaeus monodon shrimp virulence assays. However, the extracted pyomelanin pigment did confer a higher resistance to oxidative stress when incubated with wild type cells. Microarray-based transcriptomic analyses revealed that the hmgA gene deletion and subsequent pyomelanin production negatively effected the expression of 129 genes primarily involved in energy production, amino acid, and lipid metabolism, and protein translation and turnover. This transcriptional response was mediated in part by an impairment of the quorum sensing regulon as transcripts of the quorum sensing high cell density master regulator LuxR and other operonic members of this regulon were significantly less abundant in the hmgA mutant. Taken together, the results suggest that the pyomelanization of V. campbellii sufficiently impairs the metabolic activities of this organism and renders it less fit and virulent than its isogenic wild type strain.

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

    Digital Repository Service at National Institute of Oceanography (India)

    Ramamurthy, T.; Nair, G.B.

    investigations of Vibrio parahaemolyticus in oysters following outbreaks in Washington, Texas, and New York. (1997 and 1998). Appl. Envrion. Microbiol. 66, 4649- 4654. DePaola, A., Ulaszek, J., Kaysner, C. A., Tenge, B. J., Nordstrom, J. L., Wells, J., Puhr, N...-710. Andrews, L. S., DeBlanc, S., Veal, C. D., Park, D. L., 2003. Response of Vibrio parahaemolyticus O3:K6 to a hot water/ cold shock pasteurization process. Food Addit. Contam. 20, 331-334. Bag, P. K., Nandi, S., Bhadra, R. K., Ramamurthy, T., Bhattacharya, S...

  11. The ecology of Vibrio vulnificus, Vibrio cholerae, and Vibrio parahaemolyticus in North Carolina estuaries.

    Science.gov (United States)

    Blackwell, Karen Dyer; Oliver, James D

    2008-04-01

    While numerous studies have characterized the distribution and/or ecology of various pathogenic Vibrio spp., here we have simultaneously examined several estuarine sites for Vibrio vulnificus, V. cholerae, and V. parahaemolyticus. For a one year period, waters and sediment were monitored for the presence of these three pathogens at six different sites on the east coast of North Carolina in the United States. All three pathogens, identified using colony hybridization and PCR methods, occurred in these estuarine environments, although V. cholerae occurred only infrequently and at very low levels. Seventeen chemical, physical, and biological parameters were investigated, including salinity, water temperature, turbidity, dissolved oxygen, levels of various inorganic nutrients and dissolved organic carbon, as well as total vibrios, total coliforms, and E. coli. We found each of the Vibrio spp. in water and sediment to correlate to several of these environmental measurements, with water temperature and total Vibrio levels correlating highly (P<0.0001) with occurrence of the three pathogens. Thus, these two parameters may represent simple assays for characterizing the potential public health hazard of estuarine waters.

  12. Sensitivity of the vibrios to ultraviolet-radiation

    International Nuclear Information System (INIS)

    Banerjee, S.K.; Chatterjee, S.N.

    1977-01-01

    The ultraviolet-inactivation kinetics of a number of strains of Vibrio cholerae (classical), Vibrio cholerae (el tor), NAG vibrios and Vibrio parahaemolyticus were investigated. Statistical analyses revealed significant differences between any two of the four types of vibrio in respect of their sensitivity to U.V. (author)

  13. 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.

  14. White shrimp (Litopenaeus vannamei) recombinant lysozyme has antibacterial activity against Gram negative bacteria: Vibrio alginolyticus, Vibrio parahemolyticus and Vibrio cholerae.

    Science.gov (United States)

    de-la-Re-Vega, Enrique; García-Galaz, Alfonso; Díaz-Cinco, Martha E; Sotelo-Mundo, Rogerio R

    2006-03-01

    C-type lysozyme has been described as an antibacterial component of the shrimp innate defence system. We determined quantitatively the antibacterial activity of white shrimp (Litopenaeus vannamei) recombinant lysozyme against three Gram negative bacteria: Vibrio alginolyticus, Vibrio parahemolyticus and Vibrio cholerae, using a turbidimetric assay with live bacteria and differential bacterial viable count after interaction with the protein. In conclusion, the antibacterial activity of recombinant shrimp lysozyme against Vibrio sp. is at least equal to the values against the Gram positive M. luteus and more active against the shrimp pathogens V. alginolyticus and V. parahemolyticus.

  15. Vibrio chromosome-specific families

    DEFF Research Database (Denmark)

    Lukjancenko, Oksana; Ussery, David

    2014-01-01

    We have compared chromosome-specific genes in a set of 18 finished Vibrio genomes, and, in addition, also calculated the pan- and core-genomes from a data set of more than 250 draft Vibrio genome sequences. These genomes come from 9 known species and 2 unknown species. Within the finished...... chromosomes, we find a core set of 1269 encoded protein families for chromosome 1, and a core of 252 encoded protein families for chromosome 2. Many of these core proteins are also found in the draft genomes (although which chromosome they are located on is unknown.) Of the chromosome specific core protein...... families, 1169 and 153 are uniquely found in chromosomes 1 and 2, respectively. Gene ontology (GO) terms for each of the protein families were determined, and the different sets for each chromosome were compared. A total of 363 different "Molecular Function" GO categories were found for chromosome 1...

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

    Science.gov (United States)

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

    1979-01-01

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

  17. 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.

  18. Crayfish: a newly recognized vehicle for vibrio infections.

    Science.gov (United States)

    Bean, N H; Maloney, E K; Potter, M E; Korazemo, P; Ray, B; Taylor, J P; Seigler, S; Snowden, J

    1998-10-01

    We conducted a 1-year case-control study of sporadic vibrio infections to identify risk factors related to consumption of seafood products in two coastal areas of Louisiana and Texas. Twenty-six persons with sporadic vibrio infections and 77 matched controls were enrolled. Multivariate analysis revealed that crayfish (P Vibrio parahemolyticus infection (OR 9.24, P vibrio infection.

  19. Vibrio parahemolyticus bacteremia: case report.

    Science.gov (United States)

    Ng, T C; Chiang, P C; Wu, T L; Leu, H S

    1999-09-01

    Vibrio parahemolyticus (V. parahemolyticus) is a halophilic gram-negative bacillus that lives in the ocean. It is the leading cause of infectious diarrhea in Taiwan and sometimes produces soft tissue infections, but it is rarely a cause of bacteremia. There have been only 11 cases reported in the literature. Most of the cases involved a history of ingestion of seafood or exposure to seawater. In addition, those patients were all immunosuppressed, especially with leukemia and cirrhosis. We report a 60-year-old male patient with chronic hepatitis C and adrenal insufficiency. He developed V. parahemolyticus bacteremia following ingestion of seafood one week prior to admission. His condition was complicated with neck and right lower leg soft tissue infection, as well as multiple organ failure. The patient survived after intravenous ceftazidime, oral doxycycline, and surgical debridement. To our knowledge, this is the 12th reported cases on Medline, and the second bacteremic case in Taiwan. After reviewing the literature, we suggest that all patients with immunosuppressed conditions or adrenal insufficiency should eat foods that are well cooked and avoid raw seafood. Moreover, when patients who are at risk to develop fever, diarrhea, and soft tissue infection after ingestion of seafood, V. parahemolyticus infection should be suspected. All culture specimens should be inoculated on Vibrios selective media.

  20. Evidence for the introduction of the Asian red alga Neosiphonia japonica and its introgression with Neosiphonia harveyi (Ceramiales, Rhodophyta) in the Northwest Atlantic.

    Science.gov (United States)

    Savoie, Amanda M; Saunders, Gary W

    2015-12-01

    There is currently conflict in the literature on the taxonomic status of the reportedly cosmopolitan species Neosiphonia harveyi, a common red alga along the coast of Atlantic Canada and New England, USA. Neosiphonia harveyi sensu lato was assessed using three molecular markers: COI-5P, ITS and rbcL. All three markers clearly delimited three genetic species groups within N. harveyi sensu lato in this region, which we identified as N. harveyi, N. japonica and Polysiphonia akkeshiensis (here resurrected from synonymy with N. japonica). Although Neosiphonia harveyi is considered by some authors to be introduced to the Atlantic from the western Pacific, it was only confirmed from the North Atlantic suggesting it is native to this area. In contrast, Neosiphonia japonica was collected from only two sites in Rhode Island, USA, as well as from its reported native range in Asia (South Korea), which when combined with data in GenBank indicates that this species was introduced to the Northwest Atlantic. The GenBank data further indicate that N. japonica was also introduced to North Carolina, Spain, Australia and New Zealand. Despite the fact that all three markers clearly delimited N. harveyi and N. japonica as distinct genetic species groups, the ITS sequences for some N. harveyi individuals displayed mixed patterns and additivity indicating introgression of nuclear DNA from N. japonica into N. harveyi in the Northwest Atlantic. Introgression of DNA from an introduced species to a native species (i.e. 'genetic pollution') is one of the possible consequences of species introductions, and we believe this is the first documented evidence for this phenomenon in red algae. © 2015 John Wiley & Sons Ltd.

  1. Identification of capsule, biofilm, lateral flagellum, and type IV pili in Vibrio mimicus strains.

    Science.gov (United States)

    Tercero-Alburo, J J; González-Márquez, H; Bonilla-González, E; Quiñones-Ramírez, E I; Vázquez-Salinas, C

    2014-11-01

    Vibrio mimicus is a bacterium that causes gastroenteritis; it is closely related to Vibrio cholerae, and can cause acute diarrhea like cholera- or dysentery-type diarrhea. It is distributed worldwide. Factors associated with virulence (such as hemolysins, enterotoxins, proteases, phospholipases, aerobactin, and hemagglutinin) have been identified; however, its pathogenicity mechanism is still unknown. In pathogenic Vibrio species such as V. cholerae, Vibrio. parahaemolyticus and Vibrio vulnificus, capsule, biofilms, lateral flagellum, and type IV pili are structures described as essential for pathogenicity. These structures had not been described in V. mimicus until this work. We used 20 V. mimicus strains isolated from water (6), oyster (9), and fish (5) samples and we were able to identify the capsule, biofilm, lateral flagellum, and type IV pili through phenotypic tests, electron microscopy, PCR, and sequencing. In all tested strains, we observed and identified the presence of capsular exopolysaccharide, biofilm formation in an in vitro model, as well as swarming, multiple flagellation, and pili. In addition, we identified homologous genes to those described in other bacteria of the genus in which these structures have been found. Identification of these structures in V. mimicus is a contribution to the biology of this organism and can help to reveal its pathogenic behavior. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Non-lethal heat shock increased Hsp70 and immune protein transcripts but not Vibrio tolerance in the white-leg shrimp.

    Directory of Open Access Journals (Sweden)

    Nguyen Hong Loc

    Full Text Available Non-lethal heat shock boosts bacterial and viral disease tolerance in shrimp, possibly due to increases in endogenous heat shock protein 70 (Hsp70 and/or immune proteins. To further understand the mechanisms protecting shrimp against infection, Hsp70 and the mRNAs encoding the immune-related proteins prophenoloxidase (proPO, peroxinectin, penaeidin, crustin and hemocyanin were studied in post-larvae of the white-leg shrimp Litopenaeus vannamei, following a non-lethal heat shock. As indicated by RT-qPCR, a 30 min abrupt heat shock increased Hsp70 mRNA in comparison to non-heated animals. Immunoprobing of western blots and quantification by ELISA revealed that Hsp70 production after heat shock was correlated with enhanced Hsp70 mRNA. proPO and hemocyanin mRNA levels were augmented, whereas peroxinectin and crustin mRNA levels were unchanged following non-lethal heat shock. Penaeidin mRNA was decreased by all heat shock treatments. Thirty min abrupt heat shock failed to improve survival of post-larvae in a standardized challenge test with Vibrio harveyi, indicating that under the conditions of this study, L. vannamei tolerance to Vibrio infection was influenced neither by Hsp70 accumulation nor the changes in the immune-related proteins, observations dissimilar to other shrimp species examined.

  3. Non-Lethal Heat Shock Increased Hsp70 and Immune Protein Transcripts but Not Vibrio Tolerance in the White-Leg Shrimp

    Science.gov (United States)

    Loc, Nguyen Hong; MacRae, Thomas H.; Musa, Najiah; Bin Abdullah, Muhd Danish Daniel; Abdul Wahid, Mohd. Effendy; Sung, Yeong Yik

    2013-01-01

    Non-lethal heat shock boosts bacterial and viral disease tolerance in shrimp, possibly due to increases in endogenous heat shock protein 70 (Hsp70) and/or immune proteins. To further understand the mechanisms protecting shrimp against infection, Hsp70 and the mRNAs encoding the immune-related proteins prophenoloxidase (proPO), peroxinectin, penaeidin, crustin and hemocyanin were studied in post-larvae of the white-leg shrimp Litopenaeus vannamei, following a non-lethal heat shock. As indicated by RT-qPCR, a 30 min abrupt heat shock increased Hsp70 mRNA in comparison to non-heated animals. Immunoprobing of western blots and quantification by ELISA revealed that Hsp70 production after heat shock was correlated with enhanced Hsp70 mRNA. proPO and hemocyanin mRNA levels were augmented, whereas peroxinectin and crustin mRNA levels were unchanged following non-lethal heat shock. Penaeidin mRNA was decreased by all heat shock treatments. Thirty min abrupt heat shock failed to improve survival of post-larvae in a standardized challenge test with Vibrio harveyi, indicating that under the conditions of this study, L. vannamei tolerance to Vibrio infection was influenced neither by Hsp70 accumulation nor the changes in the immune-related proteins, observations dissimilar to other shrimp species examined. PMID:24039886

  4. Resistance to Antimicrobial Peptides in Vibrios

    Directory of Open Access Journals (Sweden)

    Delphine Destoumieux-Garzón

    2014-10-01

    Full Text Available Vibrios are associated with a broad diversity of hosts that produce antimicrobial peptides (AMPs as part of their defense against microbial infections. In particular, vibrios colonize epithelia, which function as protective barriers and express AMPs as a first line of chemical defense against pathogens. Recent studies have shown they can also colonize phagocytes, key components of the animal immune system. Phagocytes infiltrate infected tissues and use AMPs to kill the phagocytosed microorganisms intracellularly, or deliver their antimicrobial content extracellularly to circumvent tissue infection. We review here the mechanisms by which vibrios have evolved the capacity to evade or resist the potent antimicrobial defenses of the immune cells or tissues they colonize. Among their strategies to resist killing by AMPs, primarily vibrios use membrane remodeling mechanisms. In particular, some highly resistant strains substitute hexaacylated Lipid A with a diglycine residue to reduce their negative surface charge, thereby lowering their electrostatic interactions with cationic AMPs. As a response to envelope stress, which can be induced by membrane-active agents including AMPs, vibrios also release outer membrane vesicles to create a protective membranous shield that traps extracellular AMPs and prevents interaction of the peptides with their own membranes. Finally, once AMPs have breached the bacterial membrane barriers, vibrios use RND efflux pumps, similar to those of other species, to transport AMPs out of their cytoplasmic space.

  5. 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...

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

    Science.gov (United States)

    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.

  7. Biocompatible capped iron oxide nanoparticles for Vibrio cholerae detection

    International Nuclear Information System (INIS)

    Sharma, Anshu; Rawat, Kamla; Solanki, Pratima R; Bohidar, H B; Baral, Dinesh

    2015-01-01

    We report the studies relating to fabrication of an efficient immunosensor for Vibrio cholerae detection. Magnetite (iron oxide (Fe 3 O 4 )) 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 Fe 3 O 4 and CA-Fe 3 O 4 /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 Fe 3 O 4 , CA-Fe 3 O 4 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 (Fe 3 O 4 ) and 189.51 nm (CA-Fe 3 O 4 ) by DLS measurement. The results of electrochemical response studies of the fabricated BSA/Ab/CA-Fe 2 O 3 /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. (paper)

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

    Science.gov (United States)

    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.

  9. 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.

  10. Studies on Culture Technological Conditions of Vibrio harveyi Y91301%哈维氏弧菌Y91301菌株培养工艺的研究

    Institute of Scientific and Technical Information of China (English)

    毛芝娟; 楼丹; 吴建军

    2004-01-01

    用大黄鱼分离的哈维氏弧菌Y91301进行试验,研究接种浓度、培养瓶装液量、起始pH值、培养时间、培养温度和培养液配方等因素对菌株生长的影响,确定静置培养时最适条件为:接种母液浓度1×108~1×109cfu/ml,接种量6%~8%(相对培养瓶装液量),装液量40~50 ml/250 ml,培养液初始pH值为6.0~6.5,培养温度28℃;使用Zobell 2216E培养配方更适合大量培养和生产的需要;旋转培养能够获得更高的生物量.

  11. Effects of stocking density of the white shrimp Litopenaeus vannamei (Boone) on immunities, antioxidant status, and resistance against Vibrio harveyi in a biofloc system.

    Science.gov (United States)

    Liu, Gang; Zhu, Songming; Liu, Dezhao; Guo, Xishan; Ye, Zhangying

    2017-08-01

    Determining optimum stocking density of the white shrimp Litopenaeus vannamei (Boone) is a big concern for shrimp farmers. However, few studies have assessed the influence of stocking density on the antioxidant status, immunology, digestive enzyme activities, and growth performance of white shrimp in biofloc systems. In this study, these parameters of white shrimp in a biofloc system were compared at three stocking densities: 300 orgs m -3 as low stocking density (LD), 400 orgs m -3 as medium stocking density (MD), and 500 orgs m -3 as high stocking density (HD). The feed conversion ratio in the LD group was significantly lower than that in the MD and HD groups (P white shrimp can be seriously impaired in the HD condition (i.e., ≥500 m -3 ) in biofloc systems. These findings can be used to determine suitable stocking densities in the white shrimp farming industry using the biofloc system. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Bacteriophage interactions with marine pathogenic Vibrios

    DEFF Research Database (Denmark)

    Kalatzis, Panagiotis

    development and spreading of antibiotic resistant bacteria in the environment. Bacteriophage therapy, constitutes a potent alternative not only for treatment but also for prevention of vibriosis in aquaculture and the current thesis addresses the potential and challenges of using phages to control Vibrio...... 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...... therapy applications. Lytic phage vB_VspP_pVa5 that has been isolated against the rapidly emerging pathogen V. splendidus is also a promising candidate for phage therapy application according to its gene content and in vitro performance against its host. The genetic features of vB_VspP_pVa5 provide also...

  13. ADHERENCE AND PA THOGENICITY ASSA Y OF VIBRIO HARVE YI IN TIGER SHRIMP (PENAEUS MONODON LARVAE FOR SC REENING BIOCONTROL AGE NT

    Directory of Open Access Journals (Sweden)

    YUSMINAH H ALA

    2002-01-01

    Full Text Available Rifampicin-resistant marker w as empl oyed as a repor ter t o detect the ad here nce and colonization of V. harve yi in s hrimp larvae. Vibri o h arve yi P1B and YA3 2. 2 were isolated f rom dead s hrimp la rv ae i n Besuki, Norther n C oast of East Jav a, while V. harveyi HB3, was isolated from pris tine sea water in Pacita n, S outhe rn Coast of E ast Ja va . Vi brio metsc hnikovii used a s bioc ontr ol a gent was i solated fr om healthy shrimp larvae in Serang, We st Java. Spontane ous mutation was c onducted to generate V. harveyi P1B, YA32.2 and HB3 resistant to rifampi cin. These mutants exhibit ed similar survival ability to t heir parent al (wild type strai ns. Signifi cant larval mortalit y was o bserv ed i n s hrimp l arv ae i noculated with YA32.2 than that of larvae ino culated with P1B. Larvae inoculated with H B 3 showed the lo w est mortality. Bacterial cell c ount of Vi brio Rf* in dead larvae w ere 103-104 cells/larvae. Isolates of Vi brio metschnikovii Z an d M as biocontrol candid ates effectively r educed the growth a nd a dherence ability of YA32.2 t o shrimp larvae. Larval mort ality in rearing water inoculat ed simultaneousl y w ith YA32.2 and V. metschnikovii was lower than the one inoculated with YA32.2 alone. Therefore, Vibrio metschnikovii Z or M could be develope d as an effective pro biotic or biocontrol agent for V. harvey i in shrimp hatcheri es.

  14. Bacteriophages in the control of pathogenic vibrios

    DEFF Research Database (Denmark)

    Plaza, Nicolás; Castillo Bermúdez, Daniel Elías; Perez-Reytor, Diliana

    2018-01-01

    constitute a continuing threat for aquaculture. Moreover, the continuous use of antibiotics has been accompanied by an emergence of antibiotic resistance in Vibrio species, implying a necessity for efficient treatments. One promising alternative that emerges is the use of lytic bacteriophages; however......, there are some drawbacks that should be overcome to make phage therapy a widely accepted method. In this work, we discuss about the major pathogenic Vibrio species and the progress, benefits and disadvantages that have been detected during the experimental use of bacteriophages to their control....

  15. 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...

  16. 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...

  17. Effect of three pathogenic vibrio infections on the non-specific immunity of croaker, Pseudosciaena crocea%三种致病弧菌感染对大黄鱼非特异性免疫功能的影响

    Institute of Scientific and Technical Information of China (English)

    李思源; 葛明峰; 胡亚萍; 蔡林婷; 王国良

    2012-01-01

    Ulcer disease, which caused by the pathogens such as Vibrio alginolytkus, Vibrio harveyi and Vibrio parahaemolyticus according to the researches, is the most common, wide spreading and harmful disease of cage cultured croaker, Pseudosciaena crocea. In this research, we set 3 experimental groups with injections of 0.2 cm3 Vibrio harveyi, Vibrio alginolyticus and Vibrio parahaemolyticus, respectively, besides a control group treated with a same dose of normal saline. At the intervals ofO, 1,2,4,7, 10, 13, 16 and 20 days after infection, blood leucocytes , NBT-positive cells and lysozyme level were taken and measured for the influence on the croaker' s non-specific immunity function under the pressure of 3 pathogenic vibrio . The results show that the change of blood physiological signs in the croaker mainly occurred 1 ~ 7 days after infection. At the initial stage, each experimental group had a significantly increase in white blood cell and NBT positive cell number (p <0.05). The percentage of lymphocyte and neutrophilic granulocyte decreased with an increasing percentage of monocyte (p <0.05). Thereafter, white blood cells started to decline and the percentage of lymphocyte and neutrophilic granulocyte started to increased. Seven days after infection, the number of varied blood cells didn' t change significantly any more, and the group infected by vibrio heavily had an obvious significant change of blood physiological signs and NBT positive cell number. The activity of lysozyme in serum was higher than spleen, but both of them were first increased and then decreased, at last they went to an accordant tendency. So we conclude that the activation of stress response and non-specific immunity function' s happened at the early stage of infection. The pathogenic vibrio showed different influences, with a time lag, on the non-specific immunity.%溶藻弧菌(Vibrio alginolyticus)、哈维氏弧菌(Vibrio harveyi)、副溶血弧菌(Vibrio parahaemolyticus

  18. 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 ...

  19. 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 ...

  20. Ocean acidification alleviates low-temperature effects on growth and photosynthesis of the red alga Neosiphonia harveyi (Rhodophyta).

    Science.gov (United States)

    Olischläger, Mark; Wiencke, Christian

    2013-12-01

    This study aimed to examine interactive effects between ocean acidification and temperature on the photosynthetic and growth performance of Neosiphonia harveyi. N. harveyi was cultivated at 10 and 17.5 °C at present (~380 µatm), expected future (~800 µatm), and high (~1500 µatm) pCO2. Chlorophyll a fluorescence, net photosynthesis, and growth were measured. The state of the carbon-concentrating mechanism (CCM) was examined by pH-drift experiments (with algae cultivated at 10 °C only) using ethoxyzolamide, an inhibitor of external and internal carbonic anhydrases (exCA and intCA, respectively). Furthermore, the inhibitory effect of acetazolamide (an inhibitor of exCA) and Tris (an inhibitor of the acidification of the diffusive boundary layer) on net photosynthesis was measured at both temperatures. Temperature affected photosynthesis (in terms of photosynthetic efficiency, light saturation point, and net photosynthesis) and growth at present pCO2, but these effects decreased with increasing pCO2. The relevance of the CCM decreased at 10 °C. A pCO2 effect on the CCM could only be shown if intCA and exCA were inhibited. The experiments demonstrate for the first time interactions between ocean acidification and temperature on the performance of a non-calcifying macroalga and show that the effects of low temperature on photosynthesis can be alleviated by increasing pCO2. The findings indicate that the carbon acquisition mediated by exCA and acidification of the diffusive boundary layer decrease at low temperatures but are not affected by the cultivation level of pCO2, whereas the activity of intCA is affected by pCO2. Ecologically, the findings suggest that ocean acidification might affect the biogeographical distribution of N. harveyi.

  1. BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING

    OpenAIRE

    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 sur...

  2. AN INVESTIGATION ON PATHOGENIC VIBRIOS DISTRIBUTION IN DOMESTIC WASTEWATER

    OpenAIRE

    A. Almasi

    2005-01-01

    Municipal wastewater is one of the most important pollution sources for water supply resources. Identification and enumeration of pathogenic agents particularly pathogenic Vibrios are beneficial for controlling and prevention planning of the infectious diseases. This research was carried out to identify the distribution of the recognized pathogenic Vibrios with emphasizing on identification of Vibrio cholera in the wastewater of Kermanshah city western Iran in 2002. The method of study was cr...

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

    Directory of Open Access Journals (Sweden)

    Kevin eEsteves

    2015-07-01

    Full Text Available Vibrio parahaemolyticus and Vibrio cholerae are ubiquitous to estuarine and marine environments. These two species can induce infections in humans. Therefore understanding the structure and dynamics of non-pandemic environmental populations in temperate regions, such as Mediterranean coastal systems, is important if we are to evaluate the risks of infection to 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 conditions for V. parahaemolyticus and V. cholerae whereas the frequency of recombination events in V. cholerae was lower in low salinity. 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.

  4. 21 CFR 866.3930 - Vibrio cholerae serological reagents.

    Science.gov (United States)

    2010-04-01

    ... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3930 Vibrio... from cultured isolates derived from clinical specimens. The identification aids in the diagnosis of...

  5. Effects of Intertidal Harvest Practices on Levels of Vibrio parahaemolyticus and Vibrio vulnificus Bacteria in Oysters.

    Science.gov (United States)

    Jones, J L; Kinsey, T P; Johnson, L W; Porso, R; Friedman, B; Curtis, M; Wesighan, P; Schuster, R; Bowers, J C

    2016-08-01

    Vibrio parahaemolyticus and Vibrio vulnificus can grow rapidly in shellfish subjected to ambient air conditions, such as during intertidal exposure. In this study, levels of total and pathogenic (tdh(+) and/or trh(+)) V. parahaemolyticus and total V. vulnificus were determined in oysters collected from two study locations where intertidal harvest practices are common. Samples were collected directly off intertidal flats, after exposure (ambient air [Washington State] or refrigerated [New Jersey]), and after reimmersion by natural tidal cycles. Samples were processed using a most-probable-number (MPN) real-time PCR method for total and pathogenic V. parahaemolyticus or V. vulnificus In Washington State, the mean levels of V. parahaemolyticus increased 1.38 log MPN/g following intertidal exposure and dropped 1.41 log MPN/g after reimmersion for 1 day, but the levels were dependent upon the container type utilized. Pathogenic V. parahaemolyticus levels followed a similar trend. However, V. vulnificus levels increased 0.10 log MPN/g during intertidal exposure in Washington but decreased by >1 log MPN/g after reimmersion. In New Jersey, initial levels of all vibrios studied were not significantly altered during the refrigerated sorting and containerizing process. However, there was an increase in levels after the first day of reimmersion by 0.79, 0.72, 0.92, and 0.71 log MPN/g for total, tdh(+) and trh(+) V. parahaemolyticus, and V. vulnificus, respectively. The levels of all targets decreased to those similar to background after a second day of reimmersion. These data indicate that the intertidal harvest and handling practices for oysters that were studied in Washington and New Jersey do not increase the risk of illness from V. parahaemolyticus or V. vulnificus Vibrio parahaemolyticus and Vibrio vulnificus are the leading causes of seafood-associated infectious morbidity and mortality in the United States. Vibrio spp. can grow rapidly in shellfish subjected to ambient

  6. 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 ...

  7. Vibrio damsela as a pathogenic agent causing mortalities in cultured sea bass (Lates calcarifer)

    OpenAIRE

    Renault, Tristan; Haffner, Philippe; Malfondet, C.; Weppe, Maurice

    1994-01-01

    Vibrio anguillarum and Vibrio ordali are species frequently described as fish pathogens. Seven species of Vibrio can also be implicated in disease problems in mariculture (Toranzo 1990). sorne of Vibrios and Barja, In addition, these marine such as V. vulnificus (Tison et al.. 1982) and V. damsela (Love et al., 1981) can also cause illness homoiothermic animals

  8. 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

  9. Stress response and virulence in Vibrio anguillarum

    OpenAIRE

    Weber, Barbara

    2010-01-01

    Bacteria use quorum sensing, a cell to cell signaling mechanism mediated by small molecules that are produced by specific signal molecule synthases, to regulate gene expression in response to population density. In Vibrio anguillarum, the quorum-sensing phosphorelay channels information from three hybrid sensor kinases VanN, VanQ, CqsS that sense signal molecules produced by the synthases VanM, VanS and CqsA, onto the phosphotransferase VanU, to regulate activity of the response regulator Van...

  10. Vibrio Iron Transport: Evolutionary Adaptation to Life in Multiple Environments

    Science.gov (United States)

    Mey, Alexandra R.; Wyckoff, Elizabeth E.

    2015-01-01

    SUMMARY Iron is an essential element for Vibrio spp., but the acquisition of iron is complicated by its tendency to form insoluble ferric complexes in nature and its association with high-affinity iron-binding proteins in the host. Vibrios occupy a variety of different niches, and each of these niches presents particular challenges for acquiring sufficient iron. Vibrio species have evolved a wide array of iron transport systems that allow the bacteria to compete for this essential element in each of its habitats. These systems include the secretion and uptake of high-affinity iron-binding compounds (siderophores) as well as transport systems for iron bound to host complexes. Transporters for ferric and ferrous iron not complexed to siderophores are also common to Vibrio species. Some of the genes encoding these systems show evidence of horizontal transmission, and the ability to acquire and incorporate additional iron transport systems may have allowed Vibrio species to more rapidly adapt to new environmental niches. While too little iron prevents growth of the bacteria, too much can be lethal. The appropriate balance is maintained in vibrios through complex regulatory networks involving transcriptional repressors and activators and small RNAs (sRNAs) that act posttranscriptionally. Examination of the number and variety of iron transport systems found in Vibrio spp. offers insights into how this group of bacteria has adapted to such a wide range of habitats. PMID:26658001

  11. Vibrio elicits targeted transcriptional responses from copepod hosts.

    Science.gov (United States)

    Almada, Amalia A; Tarrant, Ann M

    2016-06-01

    Copepods are abundant crustaceans that harbor diverse bacterial communities, yet the nature of their interactions with microbiota are poorly understood. Here, we report that Vibrio elicits targeted transcriptional responses in the estuarine copepod Eurytemora affinis We pre-treated E. affinis with an antibiotic cocktail and exposed them to either a zooplankton specialist (Vibrio sp. F10 9ZB36) or a free-living species (Vibrio ordalii 12B09) for 24 h. We then identified via RNA-Seq a total of 78 genes that were differentially expressed following Vibrio exposure, including homologs of C-type lectins, chitin-binding proteins and saposins. The response differed between the two Vibrio treatments, with the greatest changes elicited upon inoculation with V. sp. F10 We suggest that these differentially regulated genes play important roles in cuticle integrity, the innate immune response, and general stress response, and that their expression may enable E. affinis to recognize and regulate symbiotic vibrios. We further report that V. sp. F10 culturability is specifically altered upon colonization of E. affinis These findings suggest that rather than acting as passive environmental vectors, copepods discriminately interact with vibrios, which may ultimately impact the abundance and activity of copepod-associated bacteria. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. 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 (oysters were highest in samples which were held for 7h in the shade (5.3 and 4.8 log MPN/g, respectively). Mean pathogenic Vp levels in oysters at initial harvest were also highest in oysters 7h in the shade (2.1 and 2.2 log MPN/g for tdh+ and trh+ Vp). Regardless of sampling location, Vibrio spp. levels were generally significantly (poysters exposed to 5h of air storage compared to the initially harvested samples. In addition, the data demonstrated that the use of layered 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

  13. [Mexican phenotype and genotype Vibrio cholerae 01].

    Science.gov (United States)

    Giono, S; Gutiérrez Cogno, L; Rodríguez Angeles, G; del Rio Zolezzi, A; Valdespino González, J L; Sepúlveda Amor, J

    1995-01-01

    This paper presents the phenotypical and genotypical characterization of 26922 Vibrio cholerae 01 strains isolated in Mexico from 1991 to 1993. All strains isolated were El Tor biovar. Strains were sensitive to antibiotics excluding furazolidone, streptomycin and sulfisoxasole to which we found resistance in 97% and we are using this characteristic as epidemiological markers. We detected a marked change in frequency of Inaba serotype from 1991, when it was dominant, with 99.5%, until 1992 when Ogawa serotype turned to be dominant with 95% of isolates. All Vibrio cholerae 01 strains, except one Ogawa strain, were to igenic, and V. choleraeno 01 were not toxigenic by ELISA, PCR and cell culture tests. Dominant ribotype was 5, but we found some strains with 6a pattern and two with ribotype 12. We are searching for ribotype 2 among hemolytic strains in order to learn if there is any relation to Gulf Coast strains prevalent in the USA, but until now we have not found any V. cholerae ribotype 2 in our isolates. Even if rapid tests are recommended for immediate diagnosis of cholera, it is necessary to continue bacterial isolation in order to have strains for phenotyping and genotyping studies that may support epidemiological analysis.

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

    Science.gov (United States)

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

    2011-05-27

    A microbiological survey was conducted to determine the levels of total and pathogenic Vibrio parahaemolyticus (Vp) and Vibrio vulnificus (Vv) in Pacific oysters (Crassostrea gigas) collected from commercial growing areas in the North Island, New Zealand. The survey was intended to be geographically representative of commercial growing areas of Pacific oysters in New Zealand, while selecting the time frame most likely to coincide with the increased abundance of pathogenic vibrio species. Vp was detected in 94.8% of oyster samples examined (n=58) with a geometric mean concentration of 99.3 MPN/g, while Vv was detected in 17.2% of oyster samples examined with a geometric mean concentration of 7.4 MPN/g. The frequency of Vp positive samples was 1.7 fold greater than reported in a study conducted three decades ago in New Zealand. Potentially virulent (tdh positive) Vp was detected in two samples (3.4%, n=58) while no trh (another virulence marker) positive samples were detected. 16S rRNA genotype could be assigned only to 58.8% of Vv isolates (8:1:1 A:B:AB ratio, n=10). There was a good agreement [98.2% of Vp (n=280) and 94.4% of Vv (n=18) isolates] between molecular tests and cultivation based techniques used to identify Vibrio isolates and there was a significant (R(2)=0.95, Pcultivation. There was no significant correlation between any of the environmental parameters tested and Vp or Vv concentrations. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Antibiotic Resistant Salmonella and Vibrio Associated with Farmed Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Sanjoy Banerjee

    2012-01-01

    Full Text Available Salmonella and Vibrio species were isolated and identified from Litopenaeus vannamei cultured in shrimp farms. Shrimp samples showed occurrence of 3.3% of Salmonella and 48.3% of Vibrio. The isolates were also screened for antibiotic resistance to oxolinic acid, sulphonamides, tetracycline, sulfamethoxazole/trimethoprim, norfloxacin, ampicillin, doxycycline hydrochloride, erythromycin, chloramphenicol, and nitrofurantoin. Salmonella enterica serovar Corvallis isolated from shrimp showed individual and multiple antibiotic resistance patterns. Five Vibrio species having individual and multiple antibiotic resistance were also identified. They were Vibrio cholerae (18.3%, V. mimicus (16.7%, V. parahaemolyticus (10%, V. vulnificus (6.7%, and V. alginolyticus (1.7%. Farm owners should be concerned about the presence of these pathogenic bacteria which also contributes to human health risk and should adopt best management practices for responsible aquaculture to ensure the quality of shrimp.

  16. Vibrio Phage KVP40 Encodes a Functional NAD+ Salvage Pathway.

    Science.gov (United States)

    Lee, Jae Yun; Li, Zhiqun; Miller, Eric S

    2017-05-01

    The genome of T4-type Vibrio bacteriophage KVP40 has five genes predicted to encode proteins of pyridine nucleotide metabolism, of which two, nadV and natV , would suffice for an NAD + salvage pathway. NadV is an apparent nicotinamide phosphoribosyltransferase (NAmPRTase), and NatV is an apparent bifunctional nicotinamide mononucleotide adenylyltransferase (NMNATase) and nicotinamide-adenine dinucleotide pyrophosphatase (Nudix hydrolase). Genes encoding the predicted salvage pathway were cloned and expressed in Escherichia coli , the proteins were purified, and their enzymatic properties were examined. KVP40 NadV NAmPRTase is active in vitro , and a clone complements a Salmonella mutant defective in both the bacterial de novo and salvage pathways. Similar to other NAmPRTases, the KVP40 enzyme displayed ATPase activity indicative of energy coupling in the reaction mechanism. The NatV NMNATase activity was measured in a coupled reaction system demonstrating NAD + biosynthesis from nicotinamide, phosphoribosyl pyrophosphate, and ATP. The NatV Nudix hydrolase domain was also shown to be active, with preferred substrates of ADP-ribose, NAD + , and NADH. Expression analysis using reverse transcription-quantitative PCR (qRT-PCR) and enzyme assays of infected Vibrio parahaemolyticus cells demonstrated nadV and natV transcription during the early and delayed-early periods of infection when other KVP40 genes of nucleotide precursor metabolism are expressed. The distribution and phylogeny of NadV and NatV proteins among several large double-stranded DNA (dsDNA) myophages, and also those from some very large siphophages, suggest broad relevance of pyridine nucleotide scavenging in virus-infected cells. NAD + biosynthesis presents another important metabolic resource control point by large, rapidly replicating dsDNA bacteriophages. IMPORTANCE T4-type bacteriophages enhance DNA precursor synthesis through reductive reactions that use NADH/NADPH as the electron donor and NAD

  17. Septicemia caused by Vibrio parahemolyticus: a case report.

    Science.gov (United States)

    Hsu, G J; Young, T; Peng, M Y; Chang, F Y; Chou, M Y

    1993-11-01

    Vibrio parahemolyticus is a halophilic marine vibrio commonly associated with outbreaks of acute gastroenteritis which also sometimes causes serious wound infection. It is an uncommon cause of septicemia. A few reports suggest that patients with chronic liver disease and leukemia are more susceptible. A case of liver cirrhosis with septicemia caused by this organism is discussed. The patient's condition rapidly deteriorated, and he died 12 hours after admission.

  18. 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

  19. Occurance and survival of Vibrio alginolyticus in Tamouda Bay (Morocco).

    Science.gov (United States)

    Sabir, M; Cohen, N; Boukhanjer, A; Ennaji, M M

    2011-10-15

    The objectives of this study were to investigate the spatial and seasonal fluctuations of Vibrio alginolyticus in marine environment of the Tamouda Bay on the Mediterranean coast of Morocco and to determine the dominant factors of the environment that govern these fluctuations. The samples (sea water, plankton, shellfish and sediment) were collected fortnightly for two years from three study sites on the coast Tamouda Bay in northern Morocco. The charge of Vibrio alginolyticus is determined by MPN method. The physicochemical parameters including temperature of sea water, pH, salinity, turbidity and chlorophyll a concentration were determined. Analysis of variance of specific variables and several principal component analyses showed that the temperature of seawater is the major determinant of seasonal distribution of Vibrio alginolyticus. The results showed a positive linear correlation between Vibrio alginolyticus and the water temperature, pH, turbidity and chlorophyll a. Similarly, there are seasonal variations and spatial of Vibrio alginolyticus in marine environment of the Tamouda bay and the highest concentrations were recorded in both years of study during the warm season whereas it was minimal during the cold season. Linear positive correlation was recorded between Vibrio alginolyticus populations in all ecological types of samples studied.

  20. VISCOSITY DICTATES METABOLIC ACTIVITY of Vibrio ruber

    Directory of Open Access Journals (Sweden)

    Maja eBoric

    2012-07-01

    Full Text Available 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 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.

  1. Intestinal Colonization Dynamics of Vibrio cholerae.

    Directory of Open Access Journals (Sweden)

    Salvador Almagro-Moreno

    2015-05-01

    Full Text Available To cause the diarrheal disease cholera, Vibrio cholerae must effectively colonize the small intestine. In order to do so, the bacterium needs to successfully travel through the stomach and withstand the presence of agents such as bile and antimicrobial peptides in the intestinal lumen and mucus. The bacterial cells penetrate the viscous mucus layer covering the epithelium and attach and proliferate on its surface. In this review, we discuss recent developments and known aspects of the early stages of V. cholerae intestinal colonization and highlight areas that remain to be fully understood. We propose mechanisms and postulate a model that covers some of the steps that are required in order for the bacterium to efficiently colonize the human host. A deeper understanding of the colonization dynamics of V. cholerae and other intestinal pathogens will provide us with a variety of novel targets and strategies to avoid the diseases caused by these organisms.

  2. Vibrio vulnificus: An Environmental and Clinical Burden

    Directory of Open Access Journals (Sweden)

    Sing-Peng Heng

    2017-05-01

    Full Text Available Vibrio vulnificus is a Gram negative, rod shaped bacterium that belongs to the family Vibrionaceae. It is a deadly, opportunistic human pathogen which is responsible for the majority of seafood-associated deaths worldwide. V. vulnificus infection can be fatal as it may cause severe wound infections potentially requiring amputation or lead to sepsis in susceptible individuals. Treatment is increasingly challenging as V. vulnificus has begun to develop resistance against certain antibiotics due to their indiscriminate use. This article aims to provide insight into the antibiotic resistance of V. vulnificus in different parts of the world as well as an overall review of its clinical manifestations, treatment, and prevention. Understanding the organism's antibiotic resistance profile is vital in order to select appropriate treatment and initiate appropriate prevention measures to treat and control V. vulnificus infections, which should eventually help lower the mortality rate associated with this pathogen worldwide.

  3. DETEKSI GEN-GEN PENYANDI FAKTOR VIRULENSI PADA BAKTERI VIBRIO

    Directory of Open Access Journals (Sweden)

    Ince Ayu Khairani Kadriah

    2011-04-01

    menggunakan isolat bakteri yang diisolasi dari budidaya udang windu di berbagai daerah di Sulawesi Selatan dan Jawa. Pada penelitian ini digunakan primer spesifik untuk mendeteksi gen-gen virulen toxR gene, hemolysin (vvh gene, dan GyrB gene dengan metode PCR. Dari 35 isolat yang diisolasi, 20 isolat terdeteksi memiliki gen virulensi dan 8 di antaranya memiliki dua gen virulen. Spesies bakteri yang memiliki gen virulen adalah: V.harveyi, V. parahaemolyticus, V. mimicus, dan V. campbelli

  4. Liquid holding recovery and photoreactivation of the ultraviolet-inactivated vibrios

    International Nuclear Information System (INIS)

    Banerjee, S.K.; Chatterjee, S.N.

    1981-01-01

    The kinetics of liquid holding recovery and photoreactivation of the ultra-violet-inactivated vibrios have been investigated. Photoreactivation was highest (about 80%) for Vibrio cholerae (classical) strains but the liquid holding recovery was highest (about 29%) for Vibrio parahemolyticus ones. Significance of the differences between any two of the four vibrio biotypes in respect of their liquid holding recovery and also photoreactivation was analysed statistically. (auth.)

  5. Mortalities of Eastern and Pacific oyster Larvae caused by the pathogens Vibrio coralliilyticus and Vibrio tubiashii.

    Science.gov (United States)

    Richards, Gary P; Watson, Michael A; Needleman, David S; Church, Karlee M; Häse, Claudia C

    2015-01-01

    Vibrio tubiashii is reported to be a bacterial pathogen of larval Eastern oysters (Crassostrea virginica) and Pacific oysters (Crassostrea gigas) and has been associated with major hatchery crashes, causing shortages in seed oysters for commercial shellfish producers. Another bacterium, Vibrio coralliilyticus, a well-known coral pathogen, has recently been shown to elicit mortality in fish and shellfish. Several strains of V. coralliilyticus, such as ATCC 19105 and Pacific isolates RE22 and RE98, were misidentified as V. tubiashii until recently. We compared the mortalities caused by two V. tubiashii and four V. coralliilyticus strains in Eastern and Pacific oyster larvae. The 50% lethal dose (LD50) of V. coralliilyticus in Eastern oysters (defined here as the dose required to kill 50% of the population in 6 days) ranged from 1.1 × 10(4) to 3.0 × 10(4) CFU/ml seawater; strains RE98 and RE22 were the most virulent. This study shows that V. coralliilyticus causes mortality in Eastern oyster larvae. Results for Pacific oysters were similar, with LD50s between 1.2 × 10(4) and 4.0 × 10(4) CFU/ml. Vibrio tubiashii ATCC 19106 and ATCC 19109 were highly infectious toward Eastern oyster larvae but were essentially nonpathogenic toward healthy Pacific oyster larvae at dosages of ≥1.1 × 10(4) CFU/ml. These data, coupled with the fact that several isolates originally thought to be V. tubiashii are actually V. coralliilyticus, suggest that V. coralliilyticus has been a more significant pathogen for larval bivalve shellfish than V. tubiashii, particularly on the U.S. West Coast, contributing to substantial hatchery-associated morbidity and mortality in recent years. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  6. Antibacterial activity of polyphenolic fraction of Kombucha against Vibrio cholerae: targeting cell membrane.

    Science.gov (United States)

    Bhattacharya, D; Ghosh, D; Bhattacharya, S; Sarkar, S; Karmakar, P; Koley, H; Gachhui, R

    2018-02-01

    The present study was undertaken to determine the mechanism of antibacterial activity of a polyphenolic fraction, composed of mainly catechin and isorhamnetin, previously isolated from Kombucha, a 14-day fermented beverage of sugared black tea, against the enteropathogen Vibrio cholerae N16961. Bacterial growth was found to be seriously impaired by the polyphenolic fraction in a dose-dependent manner. Scanning Electron Microscopy demonstrated morphological alterations in bacterial cells when exposed to the polyphenolic fraction in a concentration-dependent manner. Permeabilization assays confirmed that the fraction disrupted bacterial membrane integrity in both time- and dose-dependent manners, which were proportional to the production of intracellular reactive oxygen species (ROS). Furthermore, each of the polyphenols catechin and isorhamnetin showed the ability to permeate bacterial cell membranes by generating oxidative stress, thereby suggesting their role in the antibacterial potential of Kombucha. Thus, the basic mechanism of antibacterial activity of the Kombucha polyphenolic fraction against V. cholerae involved bacterial membrane permeabilization and morphological changes, which might be due to the generation of intracellular ROS. To the best of our knowledge, this is the first report on the investigation of antibacterial mechanism of Kombucha, which is mostly attributed to its polyphenolic content. The emergence of multidrug-resistant Vibrio cholerae strains has hindered an efficient anti-Vibrio therapy. This study has demonstrated the membrane damage-mediated antibacterial mechanism of Kombucha, a popular fermented beverage of sugared tea, which is mostly attributed to its polyphenolic content. This study also implies the exploitation of Kombucha as a potential new source of bioactive polyphenols against V. cholerae. © 2017 The Society for Applied Microbiology.

  7. Proteomics Analysis Reveals Previously Uncharacterized Virulence Factors in Vibrio proteolyticus

    Directory of Open Access Journals (Sweden)

    Ann Ray

    2016-07-01

    Full Text Available Members of the genus Vibrio include many pathogens of humans and marine animals that share genetic information via horizontal gene transfer. Hence, the Vibrio pan-genome carries the potential to establish new pathogenic strains by sharing virulence determinants, many of which have yet to be characterized. Here, we investigated the virulence properties of Vibrio proteolyticus, a Gram-negative marine bacterium previously identified as part of the Vibrio consortium isolated from diseased corals. We found that V. proteolyticus causes actin cytoskeleton rearrangements followed by cell lysis in HeLa cells in a contact-independent manner. In search of the responsible virulence factor involved, we determined the V. proteolyticus secretome. This proteomics approach revealed various putative virulence factors, including active type VI secretion systems and effectors with virulence toxin domains; however, these type VI secretion systems were not responsible for the observed cytotoxic effects. Further examination of the V. proteolyticus secretome led us to hypothesize and subsequently demonstrate that a secreted hemolysin, belonging to a previously uncharacterized clan of the leukocidin superfamily, was the toxin responsible for the V. proteolyticus-mediated cytotoxicity in both HeLa cells and macrophages. Clearly, there remains an armory of yet-to-be-discovered virulence factors in the Vibrio pan-genome that will undoubtedly provide a wealth of knowledge on how a pathogen can manipulate host cells.

  8. 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 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.

  9. 大黄鱼三种病原弧菌外膜蛋白交叉保护性抗原筛选%Selection of cross-protective antigens from outer membrane proteins of three pathogenic vibrios isolated from infected large yellow croaker (Pseudosciaena crocea)

    Institute of Scientific and Technical Information of China (English)

    张崇文; 毛芝娟; 于涟

    2012-01-01

    弧菌是海水养殖环境中常见的条件性致病菌,弧菌病的暴发给水产养殖业造成了严重损失.鉴于水生动物尤其是鱼类弧菌病的发生常常是多种(血清型或亚种)弧菌的混合感染,筛选具有潜在的交叉保护性蛋白抗原,作为制备多价疫苗或联合疫苗的侯选成分具有重要意义.文中从患病大黄鱼中分离到8株弧菌,经生理生化和分子生物学鉴定分别为6株哈维氏弧菌Vibrio harveyi,1株溶藻弧菌Vibrio alginolyticus和1株副溶血弧菌Vibrio parahaemolyticus.选择典型的不同种的弧菌为代表,提取其外膜蛋白,经SDS-PAGE和Western blotting分析,确定它们大约在45 kDa、35 kDa、22 kDa处出现了3条共同的免疫印迹条带,表明它们很有可能含有共同的能够彼此交叉保护的抗原.利用双向电泳和免疫印迹相结合的方法,借助于MALDI-TOF-MS质谱分析技术,发现溶藻弧菌V.alginolyticus的一种功能未知的孔蛋白(Porin,GenBank Accession No.ZP_01260407)和副溶血弧菌V.parahaemolyticus的一种麦芽糖孔蛋白的前体蛋白(Maltoporin precursor,GenBank Accession No.NP_801154)能够和哈维氏弧菌V.harveyi全菌多抗产生免疫反应,表明这两种蛋白可以作为3种弧菌的交叉保护性抗原,以此制备的疫苗可望对3种弧菌的感染产生交叉保护作用.%Vibrios are universal conditioned-pathogenic bacteria in marine culture environment, and the outbreak of vibrio disease resulted in a serious damage to aquaculture. Considering that vibrio disease in aquatic species, especially fishes, usually originated from mixed infection of different species (serotypes or subspecies) of vibrios, it is important to select the potential cross-protective protein antigens as candidates of polyvalent or combined vaccines. In present research, several strains of vibrios were isolated from infected large yellow croaker (Pseudosciaena crocea) and subsequently identified as six strains of V. harveyi, one V

  10. 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

  11. Cell vacuolation caused by Vibrio cholerae hemolysin.

    Science.gov (United States)

    Figueroa-Arredondo, P; Heuser, J E; Akopyants, N S; Morisaki, J H; Giono-Cerezo, S; Enríquez-Rincón, F; Berg, D E

    2001-03-01

    Non-O1 strains of Vibrio cholerae implicated in gastroenteritis and diarrhea generally lack virulence determinants such as cholera toxin that are characteristic of epidemic strains; the factors that contribute to their virulence are not understood. Here we report that at least one-third of diarrhea-associated nonepidemic V. cholerae strains from Mexico cause vacuolation of cultured Vero cells. Detailed analyses indicated that this vacuolation was related to that caused by aerolysin, a pore-forming toxin of Aeromonas; it involved primarily the endoplasmic reticulum at early times (approximately 1 to 4 h after exposure), and resulted in formation of large, acidic, endosome-like multivesicular vacuoles (probably autophagosomes) only at late times (approximately 16 h). In contrast to vacuolation caused by Helicobacter pylori VacA protein, that induced by V. cholerae was exacerbated by agents that block vacuolar proton pumping but not by endosome-targeted weak bases. It caused centripetal redistribution of endosomes, reflecting cytoplasmic alkalinization. The gene for V. cholerae vacuolating activity was cloned and was found to correspond to hlyA, the structural gene for hemolysin. HlyA protein is a pore-forming toxin that causes ion leakage and, ultimately, eukaryotic cell lysis. Thus, a distinct form of cell vacuolation precedes cytolysis at low doses of hemolysin. We propose that this vacuolation, in itself, contributes to the virulence of V. cholerae strains, perhaps by perturbing intracellular membrane trafficking or ion exchange in target cells and thereby affecting local intestinal inflammatory or other defense responses.

  12. Relationship of aquatic environmental factors with the abundance of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio mimicus and Vibrio vulnificus in the coastal area of Guaymas, Sonora, Mexico.

    Science.gov (United States)

    León Robles, A; Acedo Félix, E; Gomez-Gil, B; Quiñones Ramírez, E I; Nevárez-Martínez, M; Noriega-Orozco, L

    2013-12-01

    Members of the genus Vibrio are common in aquatic environments. Among them are V. cholerae, V. vulnificus, V. parahaemolyticus and V. mimicus. Several studies have shown that environmental factors, such as temperature, salinity, and dissolved oxygen, are involved in their epidemiology. Therefore, the main objective of this study is to determine if there is a correlation between the presence/amount of V. cholerae, V, vulnificus, V. parahaemolyticus and V. mimicus and the environmental conditions of the seawater off the coast of Guaymas, México. Quantification of all four pathogenic bacteria was performed using the most probable number method, and suspected colonies were identified by polymerase chain reaction (PCR). Correlations were found using principal component analysis. V. parahaemolyticus was the most abundant and widely distributed bacteria, followed by V. vulnificus, V. mimicus and V. cholerae. Positive correlations between V. parahaemolyticus, V. vulnificus and V. mimicus with temperature, salinity, electric conductivity, and total dissolved solids were found. The abundance of V. cholerae was mainly affected by the sampling site and not by physicochemical parameters.

  13. A pan-European ring trial to validate an International Standard for detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus in seafoods.

    Science.gov (United States)

    Hartnell, R E; Stockley, L; Keay, W; Rosec, J-P; Hervio-Heath, D; Van den Berg, H; Leoni, F; Ottaviani, D; Henigman, U; Denayer, S; Serbruyns, B; Georgsson, F; Krumova-Valcheva, G; Gyurova, E; Blanco, C; Copin, S; Strauch, E; Wieczorek, K; Lopatek, M; Britova, A; Hardouin, G; Lombard, B; In't Veld, P; Leclercq, A; Baker-Austin, C

    2018-02-10

    Globally, vibrios represent an important and well-established group of bacterial foodborne pathogens. The European Commission (EC) mandated the Comite de European Normalisation (CEN) to undertake work to provide validation data for 15 methods in microbiology to support EC legislation. As part of this mandated work programme, merging of ISO/TS 21872-1:2007, which specifies a horizontal method for the detection of V. parahaemolyticus and V. cholerae, and ISO/TS 21872-2:2007, a similar horizontal method for the detection of potentially pathogenic vibrios other than V. cholerae and V. parahaemolyticus was proposed. Both parts of ISO/TS 21872 utilized classical culture-based isolation techniques coupled with biochemical confirmation steps. The work also considered simplification of the biochemical confirmation steps. In addition, because of advances in molecular based methods for identification of human pathogenic Vibrio spp. classical and real-time PCR options were also included within the scope of the validation. These considerations formed the basis of a multi-laboratory validation study with the aim of improving the precision of this ISO technical specification and providing a single ISO standard method to enable detection of these important foodborne Vibrio spp.. To achieve this aim, an international validation study involving 13 laboratories from 9 countries in Europe was conducted in 2013. The results of this validation have enabled integration of the two existing technical specifications targeting the detection of the major foodborne Vibrio spp., simplification of the suite of recommended biochemical identification tests and the introduction of molecular procedures that provide both species level identification and discrimination of putatively pathogenic strains of V. parahaemolyticus by the determination of the presence of theromostable direct and direct related haemolysins. The method performance characteristics generated in this have been included in revised

  14. Molecular Cloning and Bioinformatics Analysis of Two-component Regulation System PhoR/PhoB fromVibrio alginolyticus HY9901%溶藻弧菌双组分调控系统PhoR/PhoB的基因克隆及生物信息学分析

    Institute of Scientific and Technical Information of China (English)

    张燕飞; 庞欢瑛; 吴灶和; 简纪常; 鲁义善

    2014-01-01

    Two-component regulatory system plays an important role in the growth and virulence of bacteria. In this study, the phosphate regulatory sensor protein (PhoR) gene and phosphate transcriptional regulatory protein(PhoB) gene ofVibrio alginolyticus strain HY9901 were cloned. Sequence analysis revealed that the length ofphoR gene was 1 299 bp, encoding a putative protein of 432 amino acids. The length ofphoB gene was 690 bp and encodes a putative protein of 299 amino acids. A phylogenetic tree was constructed by MEGA 5.0 software, indicating that the PhoR and PhoB ofV. alginolyticus showed high genetic relationship withVibrioparahaemolyticusand Vibrio harveyi. By SWISS-MODEL work-space, the three-dimensional structures of HATPase_c and REC conserved domain in the PhoR and PhoB were determined, showing that HATPase_c and REC domain contained an ATP binding site and four aspartic acid active sites, respectively, playing critical roles in regulation of growing and virulence. These results would provide a basis for the further studies on the biological function and drug development of PhoR / PhoB.%双组分调控系统(Two-component Regulatory System)在致病菌的生长及毒力调控中起重要作用。克隆溶藻弧菌(Vibrio alginolyticus)HY9901株组氨酸激酶PhoR和反应调控因子PhoB的全长基因,并对其进行生物信息学分析。序列分析结果显示,phoR(GenBank登录号:KJ958404)全长1299 bp,共编码432个氨基酸;phoB (GenBank登录号:KJ863646)全长690 bp,编码229个氨基酸。构建PhoR/PhoB的系统进化树,结果显示,溶藻弧菌PhoR/PhoB与副溶血弧菌(Vibrio parahaemolyticus)、哈氏弧菌(Vibrio harveyi)有较近亲缘关系。利用SWISS-MODEL软件,对PhoR/PhoB中两个相对保守的功能域HATPase_c和REC进行同源建模,发现HATPase_c具有1个ATP结合位点,REC具有4个天冬氨酸活性位点。

  15. 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...... patterns of the individual host isolates, key phenotypic properties related to phage susceptibility are distributed worldwide and maintained in the global Vibrio community for decades. The phage susceptibility pattern of the isolates did not show any relation to the physiological relationships obtained...... from Biolog GN2 profiles, demonstrating that similar phage susceptibility patterns occur across broad phylogenetic and physiological differences in Vibrio strains. Subsequent culture experiments with two phages and two V. anguillarum hosts demonstrated an initial strong lytic potential of the phages...

  16. Multiple enzymatic profiles of Vibrio parahaemolyticus strains isolated from oysters

    Directory of Open Access Journals (Sweden)

    Renata Albuquerque Costa

    Full Text Available The enzymatic characterization of vibrios has been used as a virulence indicator of sanitary interest. The objective of this study was to determine the enzymatic profile of Vibrio parahaemolyticus strains (n = 70 isolated from Crassostrea rhizophorae oysters. The strains were examined for the presence of gelatinase (GEL, caseinase (CAS, elastase (ELAS, phospholipase (PHOS, lipase (LIP, amilase (AML and DNase. All enzymes, except elastase, were detected in more than 60% of the strains. The most recurrent enzymatic profiles were AML + DNase + PHOS + GEL + LIP (n = 16; 22.9% and AML + CAS + DNase + PHOS + GEL + LIP (n = 21; 30%. Considering the fact that exoenzyme production by vibrios is closely related to virulence, one must be aware of the bacteriological risk posed to human health by the consumption of raw or undercooked oysters.

  17. 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.

  18. Chitovibrin: a chitin-binding lectin from Vibrio parahemolyticus.

    Science.gov (United States)

    Gildemeister, O S; Zhu, B C; Laine, R A

    1994-12-01

    A novel 134 kDa, calcium-independent chitin-binding lectin, 'chitovibrin', is secreted by the marine bacterium Vibrio parahemolyticus, inducible with chitin or chitin-oligomers. Chitovibrin shows no apparent enzymatic activity but exhibits a strong affinity for chitin and chito-oligomers > dp9. The protein has an isoelectric pH of 3.6, shows thermal tolerance, binds chitin with an optimum at pH 6 and is active in 0-4 M NaCl. Chitovibrin appears to be completely different from other reported Vibrio lectins and may function to bind V. parahemolyticus to chitin substrates, or to capture or sequester chito-oligomers. It may be a member of a large group of recently described proteins in Vibrios related to a complex chitinoclastic (chitinivorous) system.

  19. Ozone Technology for Pathogenic Bacteria of Shrimp (Vibrio sp.) Disinfection

    Science.gov (United States)

    Wulansarie, Ria; Dyah Pita Rengga, Wara; Rustamadji

    2018-03-01

    One of important marine commodities in Indonesia, shrimps are susceptible with Vibrio sp bacteria infection. That infection must be cleared. One of the technologies for disinfecting Vibrio sp. is ozone technology. In this research, Vibrio sp. is a pathogenic bacterium which infects Penaeus vannamei. Ozone technology is applied for threatening Vibrio sp. In this research, ozonation was performed in different pH. Those are neutral, acid (pH=4), and base (pH=9). The sample was water from shrimp embankment from Balai Besar Perikanan Budidaya Air Payau (BBPBAP) located in Jepara. That water was the habitat of Penaeus vannamei shrimp. The brand of ozonator used in this research was “AQUATIC”. The used ozonator in this research had 0,0325 g/hour concentration. The flow rate of sample used in this research was 2 L/minute. The ozonation process was performed in continuous system. A tank, pipe, pump, which was connected with microfilter, flowmeter and ozone generator were the main tools in this research. It used flowmeter and valve to set the flow rate scalable as desired. The first step was the insert of 5 L sample into the receptacle. Then, by using a pump, a sample supplied to the microfilter to be filtered and passed into the flow meter. The flow rate was set to 2 LPM. Furthermore, gas from ozonator passed to the flow for the disinfection of bacteria and then was recycled to the tank and the process run continuously. Samples of the results of ozonation were taken periodically from time 0, 3, 7, 12, 18, 24 to 30 minutes. The samples of the research were analyzed using Total Plate Count (TPC) test in BBPBAP Jepara to determine the number of Vibrio sp. bacteria. The result of this research was the optimal condition for pathogenic bacteria of shrimp (Vibrio sp.) ozonation was in neutral condition.

  20. 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

  1. 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

  2. Vibrio infections in Louisiana: twenty-five years of surveillance 1980-2005.

    Science.gov (United States)

    Thomas, Annu; Straif-Bourgeois, Susanne; Sokol, Theresa M; Ratard, Raoult C

    2007-01-01

    A total of 1,007 Vibrio infections were reported to the Infectious Disease Epidemiology Department at the Louisiana Office of Public Heath, between 1980 and 2005. The most common were Vibrio vulnificus (257 infections), Vibrio parahemolyticus (249 infections), and Vibrio cholerae non O1 (200 cases). Other species were much less common. Vibrio vulnificus infections, which are associated with consumption of raw seafood (particularly oysters) or contact with sea water, and severe immuno-suppression or liver disease were increasing. Septicemia and blood stream infections are the main manifestations of this infection. The number of infections due to Vibrio parahemolyticus on the other hand, causing mostly gastroenteritis, has remained stable. Vibrio cholerae infections are less common and almost always associated with consumption of partially cooked or contaminated crabs.

  3. 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.

  4. 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.

  5. Vibrio cholerae O1 secretes an extracellular matrix in response to antibody-mediated agglutination.

    Directory of Open Access Journals (Sweden)

    Danielle E Baranova

    Full Text Available Vibrio cholerae O1 is one of two serogroups responsible for epidemic cholera, a severe watery diarrhea that occurs after the bacterium colonizes the human small intestine and secretes a potent ADP-ribosylating toxin. Immunity to cholera is associated with intestinal anti-lipopolysaccharide (LPS antibodies, which are known to inhibit V. cholerae motility and promote bacterial cell-cell crosslinking and aggregation. Here we report that V. cholerae O1 classical and El Tor biotypes produce an extracellular matrix (ECM when forcibly immobilized and agglutinated by ZAC-3 IgG, an intestinally-derived monoclonal antibody (MAb against the core/lipid A region of LPS. ECM secretion, as demonstrated by crystal violet staining and scanning electron microscopy, occurred within 30 minutes of antibody exposure and peaked by 3 hours. Non-motile mutants of V. cholerae did not secrete ECM following ZAC-3 IgG exposure, even though they were susceptible to agglutination. The ECM was enriched in O-specific polysaccharide (OSP but not Vibrio polysaccharide (VPS. Finally, we demonstrate that ECM production by V. cholerae in response to ZAC-3 IgG was associated with bacterial resistant to a secondary complement-mediated attack. In summary, we propose that V. cholerae O1, upon encountering anti-LPS antibodies in the intestinal lumen, secretes an ECM (or O-antigen capsule possibly as a strategy to shield itself from additional host immune factors and to exit an otherwise inhospitable host environment.

  6. Electronics

    Science.gov (United States)

    2001-01-01

    International Acer Incorporated, Hsin Chu, Taiwan Aerospace Industrial Development Corporation, Taichung, Taiwan American Institute of Taiwan, Taipei, Taiwan...Singapore and Malaysia .5 - 4 - The largest market for semiconductor products is the high technology consumer electronics industry that consumes up...Singapore, and Malaysia . A new semiconductor facility costs around $3 billion to build and takes about two years to become operational

  7. Cell Vacuolation Caused by Vibrio cholerae Hemolysin

    Science.gov (United States)

    Figueroa-Arredondo, Paula; Heuser, John E.; Akopyants, Natalia S.; Morisaki, J. Hiroshi; Giono-Cerezo, Silvia; Enríquez-Rincón, Fernando; Berg, Douglas E.

    2001-01-01

    Non-O1 strains of Vibrio cholerae implicated in gastroenteritis and diarrhea generally lack virulence determinants such as cholera toxin that are characteristic of epidemic strains; the factors that contribute to their virulence are not understood. Here we report that at least one-third of diarrhea-associated nonepidemic V. cholerae strains from Mexico cause vacuolation of cultured Vero cells. Detailed analyses indicated that this vacuolation was related to that caused by aerolysin, a pore-forming toxin of Aeromonas; it involved primarily the endoplasmic reticulum at early times (∼1 to 4 h after exposure), and resulted in formation of large, acidic, endosome-like multivesicular vacuoles (probably autophagosomes) only at late times (∼16 h). In contrast to vacuolation caused by Helicobacter pylori VacA protein, that induced by V. cholerae was exacerbated by agents that block vacuolar proton pumping but not by endosome-targeted weak bases. It caused centripetal redistribution of endosomes, reflecting cytoplasmic alkalinization. The gene for V. cholerae vacuolating activity was cloned and was found to correspond to hlyA, the structural gene for hemolysin. HlyA protein is a pore-forming toxin that causes ion leakage and, ultimately, eukaryotic cell lysis. Thus, a distinct form of cell vacuolation precedes cytolysis at low doses of hemolysin. We propose that this vacuolation, in itself, contributes to the virulence of V. cholerae strains, perhaps by perturbing intracellular membrane trafficking or ion exchange in target cells and thereby affecting local intestinal inflammatory or other defense responses. PMID:11179335

  8. 一株海绵放线菌的分离培养、发酵条件优化及其抑制弧菌效果%Isolation, optimization of fermentation conditions and inhibition of Vibrio in a sponge-associated actinobacterium FH

    Institute of Scientific and Technical Information of China (English)

    杜英侠; 付晚涛; 李晶莹; 傅敏; 张鑫; 曲春强; 李小艺; 张菊林; 顾颖; 王选骏

    2017-01-01

    为研究放线菌对海水养殖病原弧菌(哈维弧菌Vibrio harveyi和灿烂弧菌Vibrio splendidus)的抑制效果,从繁茂膜海绵Hymeniacidon perlevis中分离筛选出了一株具有抑菌活性的放线菌(FH),并采用16S rD-NA基因序列进行分析,初步鉴定为球孢链霉菌Streptomyces globisporus;以抑菌活性为导向对放线菌FH菌株的发酵条件进行优化,并通过测量发酵沉淀菌丝体干质量,获得了FH菌株生长曲线,同时研究了发酵产物保存温度及时间对抑菌效果的影响,以及FH发酵产物在灭菌海水中对弧菌的抑制作用.结果表明:对FH菌株生长曲线测定结果显示,0~15 h为适应期,15~72 h为指数生长期,72~120 h为稳定期,120 h后为衰亡期;抑制弧菌效果试验显示,FH菌株发酵正交试验的最优条件为温度25℃、pH 7.5、 装液量30 mL(250 mL三角瓶);在正交试验最优条件下,FH菌株发酵培养72 h获得的粗提物抑制哈维弧菌和灿烂弧菌的效果最好;FH菌株发酵粗提物在4℃和-20℃下保存60 d和在室温保存7 d,其抑菌效果无显著性差异(P>0.05);在100 mL灭菌海水中,FH菌株发酵产物2 mL可有效抑制哈维弧菌和灿烂弧菌的繁殖.研究表明,海绵放线菌及其发酵产物在海水养殖领域具有潜在应用价值.%A sponge-associated actinobacterium FH with strong bioactivity was isolated from marine sponge Hy-meniacidon perlevis, and identified by 16S rDNA gene sequence analysis as Streptomyces globisporus. The inhibition of Vibrio harveyi and Vibrio splendidus by FH strain was studied and the fermentation condition of FH strain was op-timized, including influence of preservation temperature and time on the antibacterial effect of fermented product. The dry weight of mycelium revealed that the growth curve of FH strain showed a lag phase in 0-15 h, log phase in 15-72 h, stationary phase in 72-120 h and decline phase after 120 h. The optimal fermentation of FH strain was observed under

  9. 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 ...

  10. 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...

  11. 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) ...

  12. Outbreak of Vibrio parahaemolyticus Sequence Type 120, Peru, 2009.

    Science.gov (United States)

    Gonzalez-Escalona, Narjol; Gavilan, Ronnie G; Toro, Magaly; Zamudio, Maria L; Martinez-Urtaza, Jaime

    2016-07-01

    In 2009, an outbreak of Vibrio parahaemolyticus occurred in Piura, Cajamarca, Lambayeque, and Lima, Peru. Whole-genome sequencing of clinical and environmental samples from the outbreak revealed a new V. parahaemolyticus clone. All the isolates identified belonged to a single clonal complex described exclusively in Asia before its emergence in Peru.

  13. 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 serogrou...

  14. Vibrio trends in the ecology of the Venice lagoon.

    Science.gov (United States)

    Rahman, Mohammad Shamsur; Martino, Maria Elena; Cardazzo, Barbara; Facco, Pierantonio; Bordin, Paola; Mioni, Renzo; Novelli, Enrico; Fasolato, Luca

    2014-04-01

    Vibrio is a very diverse genus that is responsible for different human and animal diseases. The accurate identification of Vibrio at the species level is important to assess the risks related to public health and diseases caused by aquatic organisms. The ecology of Vibrio spp., together with their genetic background, represents an important key for species discrimination and evolution. Thus, analyses of population structure and ecology association are necessary for reliable characterization of bacteria and to investigate whether bacterial species are going through adaptation processes. In this study, a population of Vibrionaceae was isolated from shellfish of the Venice lagoon and analyzed in depth to study its structure and distribution in the environment. A multilocus sequence analysis (MLSA) was developed on the basis of four housekeeping genes. Both molecular and biochemical approaches were used for species characterization, and the results were compared to assess the consistency of the two methods. In addition, strain ecology and the association between genetic information and environment were investigated through statistical models. The phylogenetic and population analyses achieved good species clustering, while biochemical identification was demonstrated to be imprecise. In addition, this study provided a fine-scale overview of the distribution of Vibrio spp. in the Venice lagoon, and the results highlighted a preferential association of the species toward specific ecological variables. These findings support the use of MLSA for taxonomic studies and demonstrate the need to consider environmental information to obtain broader and more accurate bacterial characterization.

  15. In situ measured elimination of Vibrio cholerae from brackish water

    Czech Academy of Sciences Publication Activity Database

    Martínez-P., M. E.; Macek, Miroslav; Castro-G., M. T.

    2004-01-01

    Roč. 9, č. 1 (2004), s. 133-140 ISSN 1360-2276 R&D Projects: GA MŠk(CZ) ME 296 Grant - others:UNAM/DGAPA/PAPIT(MX) IN216796 Keywords : Vibrio cholerae * protozoan feeding * brackish water Subject RIV: EE - Microbiology, Virology Impact factor: 1.969, year: 2004

  16. 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.

  17. 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 ...

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

    African Journals Online (AJOL)

    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.

  19. Natural modulators of Vibrios in seawater and shellfish

    Science.gov (United States)

    Naturally occurring marine bacteria, Vibrio parahaemolyticus and V. vulnificus, are major threats to the safety of molluscan shellfish in the US and elsewhere. Illnesses range from mild gastrointestinal upset to septicemia and death. In studies on the uptake and persistence of V. parahaemolyticus ...

  20. 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 ...

  1. 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

  2. Microbially influenced corrosion of stainless steel by marine bacterium Vibrio natriegens: (I) Corrosion behavior

    Energy Technology Data Exchange (ETDEWEB)

    Cheng Sha; Tian Jintao [Institute of Materials Science and Engineering, Ocean University of China, Qingdao 266100 (China); Chen Shougang, E-mail: sgchen@ouc.edu.cn [Institute of Materials Science and Engineering, Ocean University of China, Qingdao 266100 (China); Lei Yanhua; Chang Xueting; Liu Tao [Institute of Materials Science and Engineering, Ocean University of China, Qingdao 266100 (China); Yin Yansheng, E-mail: yys2006@ouc.edu.cn [Institute of Materials Science and Engineering, Ocean University of China, Qingdao 266100 (China)

    2009-04-30

    The microbially influenced corrosion of stainless steel (SS) by marine bacterium Vibrio natriegens (V. natriegens) was investigated using surface analysis (atomic force microscopy (AFM), scanning electron microscopy (SEM), and energy dispersive X-ray analysis (EDXA)) and electrochemical techniques (the open circuit potential, electrochemical impedance spectroscopy (EIS), and potentiodynamic polarization curves ). AFM images corroborated the results from the EIS models which show biofilm attachment and subsequent detachment over time. The SEM images revealed the occurrence of micro-pitting corrosion underneath the biofilms on the metal surface after the biofilm removal. The presence of carbon, oxygen, phosphor and sulfur obtained from EDXA proved the formation of biofilm. The electrochemical results showed that the corrosion of SS was accelerated in the presence of V. natriegens based on the decrease in the resistance of the charge transfer resistance (R{sub ct}) obtained from EIS and the increase in corrosion current densities obtained from potentiodynamic polarization curves.

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

    OpenAIRE

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

    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 localitie...

  4. Vibrio vulnificus phage PV94 is closely related to temperate phages of V. cholerae and other Vibrio species.

    Directory of Open Access Journals (Sweden)

    Mark Pryshliak

    Full Text Available BACKGROUND: Vibrio vulnificus is an important pathogen which can cause serious infections in humans. Yet, there is limited knowledge on its virulence factors and the question whether temperate phages might be involved in pathogenicity, as is the case with V. cholerae. Thus far, only two phages (SSP002 and VvAW1 infecting V. vulnificus have been genetically characterized. These phages were isolated from the environment and are not related to Vibrio cholerae phages. The lack of information on temperate V. vulnificus phages prompted us to isolate those phages from lysogenic strains and to compare them with phages of other Vibrio species. RESULTS: In this study the temperate phage PV94 was isolated from a V. vulnificus biotype 1 strain by mitomycin C induction. PV94 is a myovirus whose genome is a linear double-stranded DNA of 33,828 bp with 5'-protruding ends. Sequence analysis of PV94 revealed a modular organization of the genome. The left half of the genome comprising the immunity region and genes for the integrase, terminase and replication proteins shows similarites to V. cholerae kappa phages whereas the right half containing genes for structural proteins is closely related to a prophage residing in V. furnissii NCTC 11218. CONCLUSION: We present the first genomic sequence of a temperate phage isolated from a human V. vulnificus isolate. The sequence analysis of the PV94 genome demonstrates the wide distribution of closely related prophages in various Vibrio species. Moreover, the mosaicism of the PV94 genome indicates a high degree of horizontal genetic exchange within the genus Vibrio, by which V. vulnificus might acquire virulence-associated genes from other species.

  5. Vibrio Parahemolyticus in the Wastewater of Kermanshah City

    Directory of Open Access Journals (Sweden)

    Ali Almasi

    2005-11-01

    Full Text Available آب و فاضلاب                                                                                                                                                                                                               شماره 51- سال 1383     Municipal wastewater is one of the most important pollution sources for water supply resources. Soil, vegetable, and food material are exposed as well. Identification and enumeration of pathogenic agents particularly pathogenic Vibrios are beneficial for control and prevention planning of the infectious diseases. This research carried out to identify the distribution of the recognized pathogenic Vibrios emphasizing on identification of Vibrio cholerain the wastewater of city of Kermanshah in 2001. Population of city of Kermanshah was estimated over 713000 and produced wastewater was approximately 150 l/cap/d. The method of study was cross-sectional descriptive. Sampling procedure was adopted from standard Methods for the Examination of water and wastewater, and the method for Vibrios identification was according to finegold 1990. There were 8 discharge outlet domestic wastewaters, which had been chosen as sampling sites. Samples were collected weekly in randomized manner in day time. Although 288 samples should be collected statistically, 339 samples were collected and analyzed. The results indicated that site 7 with 5 positives, sites 4 and 8 with 3 positives, site 5 with 2 postitives and sites 2, 3 and 6 with one positive suspected to vibrio pathogens. However, not any Vibrio detected in site 1. The most positive samples were seen in spring, late summer and early autumn. The positive results were detected in May, June, September, and October. Among samples which have been detected as a

  6. MR findings of infectious myositis caused by vibrio vulnificus: case report

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Joon Ho; Na, Jae Boem [Gyeongsang National University College of Medicine, Jinju (Korea, Republic of)

    2003-03-01

    Vibrio vulnificus infection is a fatal disease occurring after the consumption of seafood in patients with underlying liver disease. Inflammation of the skin, subcutanous fat and fascia disseminates from the lower extremity to the trunk and upper extremity. Infection myositis caused by vibrio vulnificus is rare, and its MR findings have not been reported. We report these in a case of infectious myositis caused by vibrio vulnificus involving both lower extremities.

  7. MR findings of infectious myositis caused by vibrio vulnificus: case report

    International Nuclear Information System (INIS)

    Lee, Joon Ho; Na, Jae Boem

    2003-01-01

    Vibrio vulnificus infection is a fatal disease occurring after the consumption of seafood in patients with underlying liver disease. Inflammation of the skin, subcutanous fat and fascia disseminates from the lower extremity to the trunk and upper extremity. Infection myositis caused by vibrio vulnificus is rare, and its MR findings have not been reported. We report these in a case of infectious myositis caused by vibrio vulnificus involving both lower extremities

  8. Efek Antibakteri Ekstrak Daun Mimba (Azadirachta indica A. Juss) terhadap Bakteri Vibrio algynoliticus Secara In Vitro

    OpenAIRE

    Uli Ayini; Siti Harnina B.; Titis Candra Dewi

    2014-01-01

    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,...

  9. Emergent Patterns of Diversity and Dynamics in Natural Populations of Planktonic Vibrio Bacteria

    Science.gov (United States)

    2005-06-01

    1973. Ecology of Vibrio parahemolyticus in mixed-template amplifications: formation, consequences and elimination by Chesapeake Bay. J. Bacteriol. 113...Science 1930 and Engineering DOCTORAL DISSERTATION Emergent Patterns of Diversity and Dynamics in Natural Populations of Planktonic Vibrio Bacteria by...DYNAMICS IN NATURAL POPULATIONS OF PLANKTONIC VIBRIO BACTERIA by Janelle Ren6e Thompson B.S. Biological Sciences, Stanford University 1998 M.S

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

    OpenAIRE

    Fairweather Morgan G; Krishnan Sujatha; Fernando Rajeev R; Ericsson Charles D

    2011-01-01

    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 ...

  11. Hemolytic and urease activities in vibrios isolated from fresh and frozen oysters

    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.

  12. Role and regulation of the orphan AphA protein of quorum sensing in pathogenic Vibrios.

    Science.gov (United States)

    Lu, Renfei; Osei-Adjei, George; Huang, Xinxiang; Zhang, Yiquan

    2018-03-01

    Quorum sensing (QS), a cell-to-cell communication process, is widely distributed in the bacterial kingdom. Bacteria use QS to control gene expression in response to cell density by detecting the signal molecules called autoinducers. AphA protein is the master QS regulator of vibrios operating at low cell density. It regulates the expression of a variety of genes, especially those encoding virulence factors, flagella/motility and biofilm formation. The role and regulation of AphA in vibrios, especially in human pathogenic vibrios, are summarized in this review. Clarification of the roles of AphA will help us to understand the pathogenesis of vibrios.

  13. Impact of PFOS on Growth and ECP Gene of Vibrio harveyi%PFOS对哈维氏弧菌生长及外毒素基因的影响

    Institute of Scientific and Technical Information of China (English)

    舒波; 黄德贵; 关万春; 潘长旺; 常彦祥

    2011-01-01

    探讨了全氟辛烷磺酸盐(PROS)对哈维氏弧菌密度及外毒素基因表达的影响.用含有不同浓度PFOS(5、50、500 mmol/L)的培养基培养哈维氏弧菌,分别在6、15、24 h取适量菌液,经4 000 r/min离心,磷酸盐缓冲液(PBS)洗涤3次,制成PBS菌悬液.采用实时荧光定量PCR检测其胞外蛋白酶、溶血素及保守序列16 SrRNA基因在不同培养时间的拷贝数.结果表明,仅5mmol/L PFOS对细菌生长有明显的抑制作用(P<0.01),其它两浓度均无明显影响.3个浓度的PFOS都使胞外蛋白酶和溶血素基因的拷贝数有明显增加(P<0.01),而16 SrRNA的基因拷贝数仅受低浓度的5 mmol/L PFOS影响而有所增加(P<0.01).同时,溶血素基因与16 SrRNA类似,对低浓度PFOS更为易感.提示PFOS对哈维氏弧菌的生长及基因有一定影响,尤以低浓度PFOS为甚.

  14. 哈维氏弧菌黑鲷分离株BK-1培养条件优化研究%Studies on the Optimal Culture Conditions of Vibrio harveyi BK-1 from Sparus macrocephalus

    Institute of Scientific and Technical Information of China (English)

    潘晓艺; 沈锦玉; 尹文林; 曹铮; 马海其; 常抗美

    2005-01-01

    从患病黑鲷Sparus macrocephalus 的肾脏分离到一致病菌株BK-1,经鉴定为哈维氏弧菌.对哈维氏弧菌BK-1株的最佳生长条件及培养基优化进行了测定.结果表明:不同的培养条件和培养基成份均会影响其产量.哈维氏弧菌最适宜生长条件为:盐度为2%、pH8、温度为30℃;最佳培养基成份为:蛋白胨0.5%,牛肉膏0.75%,甘油0.05%,CuSO40.15 mg/L,CaCl2 0.01 g/L.

  15. Studies on the Optimal Culture Conditions of Vibrio harveyi GYC1108-1 from Pseudosciaena crocea%大黄鱼细菌性病原哈维氏弧菌培养特性的研究

    Institute of Scientific and Technical Information of China (English)

    沈锦玉; 潘晓艺; 尹文林; 郝贵杰; 曹铮

    2009-01-01

    弧菌是危害水产养殖动物的最为严重的病原之一.从患病大黄鱼肝脏分离致病的细菌哈维氏弧菌GYC1108-1为材料,对哈维氏弧菌GYC1108-1株的最佳生长条件及培养基优化进行了测定.综合考察不同的培养温度、培养时间、培养基盐度、培养基的pH值对细菌生长的影响,并采用正交实验法,对培养基配方进行优化.结果表明:不同的培养温度、培养时间、培养基盐度、培养基pH值等均会影响细菌的产量.GYC1108-1适宜生长的盐度为1%-5%、pH为5~9.5、温度为15~35℃、在培养基中添加硫酸铜、硫酸氨会明显促进生长;最佳培养条件及培养基成份为:NaCl 2%,pH8.0,温度30℃,蛋白胨1.0%、牛肉膏0.75%、CuSO4 0.3mg/L、(NH4)2SO4 0.1g/L.

  16. The Optimal Culture Conditions of Bacterium Vibrio harveyi WG1702 from Clam Meretrix meretrix%哈氏弧菌文蛤分离株WG1702培养条件优化研究

    Institute of Scientific and Technical Information of China (English)

    刘连生; 闫茂仓; 赵海泉; 陈倩影; 马爱敏; 林志华

    2010-01-01

    于2007年9月在江苏吕泗文蛤发病高峰期,从发病文蛤体内和养殖水体中分离到1株哈氏弧菌 WG1702,培养条件优化试验结果表明,菌株WG1702在32 h时达到生长高峰,最适温度为28 ℃,最佳初始pH为7,最佳盐度为40,最佳装液量为60 ml/150 ml,促进生长的金属离子是NH_4~+、Fe~(2+) 、Fe~(3+) ,最佳C、N源为葡萄糖和蛋白胨;pH、温度、不同菌龄正交试验证明最佳组合为pH 7.11,温度28 ℃,菌龄24 h,通过方差分析结果可以看出三因素中pH对生长影响显著( P <0.05).

  17. Investigation of household contamination of Vibrio cholerae in Bangladesh

    DEFF Research Database (Denmark)

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

    . cholerae El Tor strain N16961, showed hemolysis and proteolysis activity but none of them exhibited any hemagglutinin activity on human erythrocytes. The study findings indicate that V. cholerae contamination is mostly originated in and around kitchen area rather than latrine area. Contaminated food...... and water supply may be the reason behind this relatively high presence of virulence factors in food plates and water pots. Direct exposure routes of disease transmission should be a major consideration in cholera prevention policies. Investigation of household contamination of Vibrio cholerae in Bangladesh......The role of in-house transmission on the incidence of Vibrio cholerae, the deadly waterborne pathogen, is still not developed. The aim of the current study was to investigate possible contamination routes in household domain for effective cholera control in Bangladesh. To examine the prevalence...

  18. Vibrio bacteria in raw oysters: managing risks to human health.

    Science.gov (United States)

    Froelich, Brett A; Noble, Rachel T

    2016-03-05

    The human-pathogenic marine bacteria Vibrio vulnificus and V. parahaemolyticus are strongly correlated with water temperature, with concentrations increasing as waters warm seasonally. Both of these bacteria can be concentrated in filter-feeding shellfish, especially oysters. Because oysters are often consumed raw, this exposes people to large doses of potentially harmful bacteria. Various models are used to predict the abundance of these bacteria in oysters, which guide shellfish harvest policy meant to reduce human health risk. Vibrio abundance and behaviour varies from site to site, suggesting that location-specific studies are needed to establish targeted risk reduction strategies. Moreover, virulence potential, rather than simple abundance, should be also be included in future modeling efforts. © 2016 The Author(s).

  19. The squid-Vibrio symbioses: from demes to genes.

    Science.gov (United States)

    Kimbell, Jennifer R; McFall-Ngai, Margaret J

    2003-04-01

    The monospecific light organ association between the Hawaiian sepiolid squid Euprymna scolopes and the marine luminous bacterium Vibrio fischeri has been used as a model for the study of the most common type of coevolved animal-bacterial interaction; i.e., the association of Gram-negative bacteria with the extracellular apical surfaces of polarized epithelia. Analysis of the squid-vibrio symbiosis has ranged from characterizations of the harvesting mechanisms by which the host ensures colonization by the appropriate symbiont to identification of bacteria-induced changes in host gene expression that accompany the establishment and maintenance of the relationship. Studies of this model have been enhanced by extensive collaboration with microbiologists, who are able to manipulate the genetics of the bacterial symbiont. The results of our studies have indicated that initiation and persistence of the association requires a complex, reciprocal molecular dialogue between these two phylogenetically distant partners.

  20. Characterization of the secretomes of two vibrios pathogenic to mollusks.

    Directory of Open Access Journals (Sweden)

    Stéphanie Madec

    Full Text Available 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.

  1. 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...

  2. Impacts of Climatic Variability on Vibrio parahaemolyticus Outbreaks in Taiwan

    OpenAIRE

    Hsin-I Hsiao; Man-Ser Jan; Hui-Ju Chi

    2016-01-01

    This study aimed to investigate and quantify the relationship between climate variation and incidence of Vibrio parahaemolyticus in Taiwan. Specifically, seasonal autoregressive integrated moving average (ARIMA) models (including autoregression, seasonality, and a lag-time effect) were employed to predict the role of climatic factors (including temperature, rainfall, relative humidity, ocean temperature and ocean salinity) on the incidence of V. parahaemolyticus in Taiwan between 2000 and 201...

  3. Temperature affects species distribution in symbiotic populations of Vibrio spp.

    Science.gov (United States)

    Nishiguchi, M K

    2000-08-01

    The genus Sepiola (Cephalopoda: Sepiolidae) contains 10 known species that occur in the Mediterranean Sea today. All Sepiola species have a light organ that contains at least one of two species of luminous bacteria, Vibrio fischeri and Vibrio logei. The two Vibrio species coexist in at least four Sepiola species (S. affinis, S. intermedia, S. ligulata, and S. robusta), and their concentrations in the light organ depend on changes in certain abiotic factors, including temperature. Strains of V. fischeri grew faster in vitro and in Sepiola juveniles when they were incubated at 26 degrees C. In contrast, strains of V. logei grew faster at 18 degrees C in culture and in Sepiola juveniles. When aposymbiotic S. affinis or S. ligulata juveniles were inoculated with one Vibrio species, all strains of V. fischeri and V. logei were capable of infecting both squid species at the optimum growth temperatures, regardless of the squid host from which the bacteria were initially isolated. However, when two different strains of V. fischeri and V. logei were placed in direct competition with each other at either 18 or 26 degrees C, strains of V. fischeri were present in sepiolid light organs in greater concentrations at 26 degrees C, whereas strains of V. logei were present in greater concentrations at 18 degrees C. In addition to the competition experiments, the ratios of the two bacterial species in adult Sepiola specimens caught throughout the season at various depths differed, and these differences were correlated with the temperature in the surrounding environment. My findings contribute additional data concerning the ecological and environmental factors that affect host-symbiont recognition and may provide insight into the evolution of animal-bacterium specificity.

  4. Distribution ofVibrio cholerae in two Florida estuaries.

    Science.gov (United States)

    Hood, M A; Ness, G E; Rodrick, G E; Blake, N J

    1983-04-01

    The distribution ofVibrio cholerae was examined in 2 Florida estuaries, Apalachicola and Tampa Bay.Vibrio cholerae serotype non-01 was the most abundant serotype, being isolated from 45% of the oyster samples, 30% of the sediments, 50% of the waters, and 75% of the blue crabs.Vibrio cholerae serotype 01 was isolated from only one oyster sample. Strong linear correlations betweenV. cholerae and temperature, salinity, or the other physical/chemical parameters measured,Escherichia coli, or fecal coliforms were not observed, but a range of temperatures and salinities appeared relevant to the distribution of the organism. The organism was present in the highest concentrations when salinities were 10‰-25‰ and temperatures were 20‡C-35‡C.In vitro growth curves of 95V. cholerae environmental isolates further supported that 10‰-25‰ was an ideal salinity range for the organisms. The results suggest thatV. cholerae is a widely distributed organism in the nutrient-rich warm waters of the Gulf Coast estuaries.

  5. [Characterization of haemolysis of the Vibrio parahaemolyticus no.93].

    Science.gov (United States)

    Su, S C; Lee, C Y

    1997-02-01

    Vibrio parahaemolyticus is a causative bacterium of food poisoning, and the haemolysin produced by this organism has been considered as one of the important virulence factors. In order to understand the pathogenic mechanism of this bacterium, the characteristics of haemolysin from Vibrio parahaemolyticus isolated from Taiwan were studied. One of the clinical strains, V. parahaemolyticus No.93, presents a weak hemolytic zone on 7% NaCl-Wagatsuma medium. The DNA hybridization results show that V. parahemolyticus has neither tdh nor trh gene. V. parahaemolyticus No.93 shows obviously hemolytic zone on 3%-NaCl Wagatsuma medium (human blood). The crude extracellular protein of V. parahaemolyticus No. 93 was evaluated for its heat tolerance and enzyme activities by media assay. The results show that this crude extracellular protein is thermolabile. The crude extracellular protein of V. parahaemolyticus No.93 was analyzed on 10% SDS-PAGE and an apparent band of 64 kDa protein was observed. Furthermore, the crude extracellular protein was analyzed by running gelatin-SDS-PAGE and hemoglobin-SDS-PAGE, and three clear zones on 62 kDa, 52 kDa and 41 kDa were observed on both SDS-PAGEs. Thus we propose that the crude extracellular protein of the V. parahaemolyticus No.93 can degrade gelatin as well as hemoglobin. Whether these protease being the virulence factors of Vibrio parahaemolyticus No.93 needs to be further studied.

  6. 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

    2016-05-24

    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 Vibrio after 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.

  7. Label-free electrochemical immunosensor based on cerium oxide nanowires for Vibrio cholerae O1 detection

    International Nuclear Information System (INIS)

    Tam, Phuong Dinh; Thang, Cao Xuan

    2016-01-01

    This paper developed a label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application. The CeO 2 nanowires were synthesized by hydrothermal reaction. The immobilization of Anti-V. cholerae O1 onto CeO 2 nanowire-deposited sensor was performed via an amino ester, which was created by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and sulfo-N-hydroxysuccinimide. The electrochemical responses of the immunosensor were studied by electrochemical impedance spectroscopy with [Fe (CN) 6 ] 3−/4− as redox probe. A linear response in electron transfer resistance for cell of V. cholerae O1 concentration was found in the range of 1.0 × 10 2 CFU/mL to 1.0 × 10 4 CFU/mL. The detection limit of the immunosensor was 1.0 × 10 2 CFU/mL. The immunosensor sensitivity was 56.82 Ω/CFU·mL −1 . Furthermore, the parameters affecting immunosensor response were also investigated, as follows: pH value, immunoreaction time, incubation temperature, and anti-V. cholerae O1 concentration. - Highlights: • A label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application was developed. • A linear response was found in the range of 1.0 × 10 2 CFU/mL to 1.0 × 10 4 CFU/mL. • The detection limit of the immunosensor was 1.0 × 10 2 CFU/mL. • The immunosensor sensitivity was 56.82 Ω/CFU.mL −1 .

  8. Label-free electrochemical immunosensor based on cerium oxide nanowires for Vibrio cholerae O1 detection

    Energy Technology Data Exchange (ETDEWEB)

    Tam, Phuong Dinh, E-mail: phuongdinhtam@gmail.com; Thang, Cao Xuan, E-mail: thang.caoxuan@hust.edu.vn

    2016-01-01

    This paper developed a label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application. The CeO{sub 2} nanowires were synthesized by hydrothermal reaction. The immobilization of Anti-V. cholerae O1 onto CeO{sub 2} nanowire-deposited sensor was performed via an amino ester, which was created by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and sulfo-N-hydroxysuccinimide. The electrochemical responses of the immunosensor were studied by electrochemical impedance spectroscopy with [Fe (CN) {sub 6}] {sup 3−/4−} as redox probe. A linear response in electron transfer resistance for cell of V. cholerae O1 concentration was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. The immunosensor sensitivity was 56.82 Ω/CFU·mL{sup −1}. Furthermore, the parameters affecting immunosensor response were also investigated, as follows: pH value, immunoreaction time, incubation temperature, and anti-V. cholerae O1 concentration. - Highlights: • A label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application was developed. • A linear response was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. • The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. • The immunosensor sensitivity was 56.82 Ω/CFU.mL{sup −1}.

  9. 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.

  10. Genome Sequence of Vibrio cholerae Strain O1 Ogawa El Tor, Isolated in Mexico, 2013

    OpenAIRE

    Díaz-Quiñonez, José Alberto; Hernández-Monroy, Irma; López-Martínez, Irma; Ortiz-Alcántara, Joanna; González-Durán, Elizabeth; Ruiz-Matus, Cuitláhuac; Kuri-Morales, Pablo; Ramírez-González, José Ernesto

    2014-01-01

    We present the draft genome sequence of Vibrio cholerae InDRE 3140 recovered in 2013 during a cholera outbreak in Mexico. The genome showed the Vibrio 7th pandemic islands VSP1 and VSP2, the pathogenic islands VPI-1 and VPI-2, the integrative and conjugative element SXT/R391 (ICE-SXT), and both prophages CTXφ and RS1φ.

  11. 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

  12. 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.

  13. Vibrio Pathogens: A Public Health Concern in Rural Water Resources in Sub-Saharan Africa

    Science.gov (United States)

    Osunla, Charles A.

    2017-01-01

    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. PMID:28991153

  14. 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 ...

  15. 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

  16. Vibrios on the half shell: what the walrus and the carpenter didn't know.

    Science.gov (United States)

    Blake, P A

    1983-10-01

    At least nine Vibrio species have been associated with disease in the United States. Vibrio fluvialis, V. hollisae, V. mimicus, and V. parahaemolyticus cause diarrheal diseases, but may also be encountered in extraintestinal infections such as wound and ear infections, septicemia, and cholecystitis. Vibrio alginolyticus, V. damsela, V. metschnikovii, and V. vulnificus primarily cause extraintestinal disease. Toxigenic V. cholerae O1 is the cause of epidemic cholera, whereas nontoxigenic V. cholerae O1 and non-O1 V. cholerae have been associated with both diarrheal and extraintestinal diseases. Most reports of vibrio infections have come from states along the Atlantic Ocean and Gulf of Mexico and from Hawaii, and most of the infections have occurred during summer and fall. Wound and ear infections have occurred after exposure to salty or brackish water or to drippings from raw seafoods. Foodborne vibrio infections are almost all caused by seafoods, especially oysters eaten raw. Thorough cooking and careful handling will render seafoods safe for consumption.

  17. 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.

  18. 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...

  19. Temperature-dependent inhibition of opportunistic Vibrio pathogens by native coral commensal bacteria.

    Science.gov (United States)

    Frydenborg, Beck R; Krediet, Cory J; Teplitski, Max; Ritchie, Kim B

    2014-02-01

    Bacteria living within the surface mucus layer of corals compete for nutrients and space. A number of stresses affect the outcome of this competition. The interactions between native microorganisms and opportunistic pathogens largely determine the coral holobiont's overall health and fitness. In this study, we tested the hypothesis that commensal bacteria isolated from the mucus layer of a healthy elkhorn coral, Acropora palmata, are capable of inhibition of opportunistic pathogens, Vibrio shiloi AK1 and Vibrio coralliilyticus. These vibrios are known to cause disease in corals and their virulence is temperature dependent. Elevated temperature (30 °C) increased the cell numbers of one commensal and both Vibrio pathogens in monocultures. We further tested the hypothesis that elevated temperature favors pathogenic organisms by simultaneously increasing the fitness of vibrios and decreasing the fitness of commensals by measuring growth of each species within a co-culture over the course of 1 week. In competition experiments between vibrios and commensals, the proportion of Vibrio spp. increased significantly under elevated temperature. We finished by investigating several temperature-dependent mechanisms that could influence co-culture differences via changes in competitive fitness. The ability of Vibrio spp. to utilize glycoproteins found in A. palmata mucus increased or remained stable when exposed to elevated temperature, while commensals' tended to decrease utilization. In both vibrios and commensals, protease activity increased at 30 °C, while chiA expression increased under elevated temperatures for Vibrio spp. These results provide insight into potential mechanisms through which elevated temperature may select for pathogenic bacterial dominance and lead to disease or a decrease in coral fitness.

  20. Incidence of Vibrio cholerae and related vibrios in a coastal lagoon and seawater influenced by lake discharges along an annual cycle.

    Science.gov (United States)

    Garay, E; Arnau, A; Amaro, C

    1985-08-01

    Most probable numbers of Vibrio cholerae and related vibrios were determined in Albufera Lake, Valencia, Spain, and in coastal waters under the influence of the lake discharges over the course of an annual cycle. The influence of temperature, kind of water, and characteristics of the different sampling sites on the numbers of vibrios recovered was evaluated. Maximum recovery of vibrios reached 10(3)/ml in both types of waters analyzed. V. cholerae numbers reached 10(3)/ml in the lake and 10(2) in one of the coastal sites. Frequently during the warm season, all vibrios isolated were identified as V. cholerae. Occasionally, no V. cholerae was recovered. The recovery of vibrios was significantly influenced by the temperature of the water and the type of water analyzed. Most of the V. cholerae isolates were included in Heiberg groups I and II, and nearly 50% of the strains used chitin as sole carbon source. Indole was not produced by 100% of the strains. All strains tested were non-O1 serovars.

  1. Vibrio cholerae Classical Biotype Strains Reveal Distinct Signatures in Mexico

    OpenAIRE

    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

    2012-01-01

    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 199...

  2. Environmental determinants of Vibrio parahaemolyticus in the Chesapeake Bay.

    Science.gov (United States)

    Davis, Benjamin J K; Jacobs, John M; Davis, Meghan F; Schwab, Kellogg J; DePaola, Angelo; Curriero, Frank C

    2017-08-25

    Vibrio parahaemolyticus naturally-occurs in brackish and marine waters and is one of the leading causes of seafood-borne illness. Previous work studying the ecology of V. parahaemolyticus is often limited in geographic extent and lacking a full range of environmental measures. This study used a unique, large dataset of surface water samples in the Chesapeake Bay ( n =1,385) collected from 148 monitoring stations from 2007 to 2010. Water was analyzed for over 20 environmental parameters with additional meteorological and surrounding land use data. V. parahaemolyticus -specific genetic markers thermolabile hemolysin ( tlh ), thermostable direct hemolysin ( tdh ), and tdh -related hemolysin ( trh ) were assayed using quantitative PCR (qPCR), and interval-censored regression models with non-linear effects were estimated to account for limits of detection and quantitation. tlh was detected in 19.6% of water samples; tdh or trh markers were not detected. Results confirmed previously reported positive associations for V. parahaemolyticus abundance with temperature and turbidity and negative associations with high salinity (> 10-23‰). Furthermore, the salinity relationship was determined to be a function of both low temperature and turbidity, with an increase of either nullifying the high salinity effect. Associations with dissolved oxygen and phosphate also appeared stronger when samples were taken nearby human developments. Renewed focus on the V. parahaemolyticus ecological paradigm is warranted to protect public health. Importance Vibrio parahaemolyticus is one of the leading causes of seafood-borne illness in the United States and across the globe. Exposure is often through consuming raw or undercooked shellfish. Given the natural presence of the bacterium in the marine environment, improved understanding of its environmental determinants is necessary for future preventative measures. This analysis of environmental Vibrio parahaemolyticus is one of only a few that

  3. 副溶血弧菌一种糖蛋白酶(复苏促进因子)在大肠杆菌中的表达及性质%Expression and Characterization of a Resuscitation-Promoting Factor Like Glycoprotease Gene from Vibrio parahaemolyticus

    Institute of Scientific and Technical Information of China (English)

    赵明君; 贾俊涛; 陈吉祥

    2012-01-01

    A resuscitation-promoting factor like glycoprotease gene was cloned from the chromosomal DNA of pathogenic Vibrio parahaemolyticus 8621. 4 by PCR amplification. The ORF of the glycoprotease gene consisted of 702 base pairs, encoding a polypeptide of 233 amino acids. The homologies of the nucle-otide sequence with those of other V. Parahaemolyticus stains were 100%. The similarities of the amino acid sequence with glycoproteases of Vibrio harveyi HY01, Vibrio sp. Ex25, Vibrio coralliilyticus , Vibrio mimicus and Vibrio salmonicida were from 67 % to 92 %. The glycoprotease gene was then subcloned into pET28a and expressed in BL21 (DE3). The polyclonal antibody was prepared by immunization of rabbits with the expressed protein, which was purified by Ni ( + )-affinity chromatography. A specific protein band of 27 kDa was detected in the whole-cell lysate preparation of normal cultural cells and 45 days culture cells of V. Parahaemolyticus 8621. 4 at low temperature conditions by western-blotting analysis , and one additional band could be detected in the VBNC cells. These results may be of significance in evaluating the role of Gcp in entering into the VBNC state under starvation stresses and recovering from the VBNC state.%用PCR方法从副溶血弧菌8621.4基因组中扩增出1种细胞复苏促进因子家族的糖蛋白酶(Glycoprotease,Gcp)基因,核酸序列分析表明其含有完整的gcp基因开放阅读框,编码233个氨基酸组成的蛋白质.核酸序列与副溶血弧菌糖蛋白酶家族基因的序列相似性为100%.其氨基酸序列与哈维氏弧菌HY01、弧菌Ex25、溶珊瑚弧菌、拟态弧菌和杀鲑弧菌LFI1238等糖蛋白酶的序列相似性为67%~92%.将该基因克隆到表达载体pET28a,在大肠杆菌BL21 (DE3)中诱导表达,用Ni琼脂糖亲和柱层析纯化的蛋白为单一条带,利用纯化的Gcp免疫新西兰大白兔制备特异性抗体,Western-Blotting分析发现正常生长及活的非可培养状态(VBNC)

  4. Preliminary study of transplanting as a process for reducing levels of Vibrio vulnificus and Vibrio parahaemolyticus in shellstock oysters.

    Science.gov (United States)

    Walton, William C; Nelson, Chris; Hochman, Mona; Schwarz, John

    2013-01-01

    Increasingly strict standards for harvest of oysters for the raw, half-shell market (designated as "white tag") should increase the proportion of oysters not meeting these standards (designated as "green tag"). Transplanting of green tag oysters into highsalinity waters (>20 practical salinity units) was explored as a means of returning Vibrio vulnificus and Vibrio parahaemolyticus levels to levels present on initial harvest. In summer 2011, oysters originally harvested in Louisiana were transplanted on two separate occasions (n = 2) to two sites in Mississippi Sound, AL: Sandy Bay and Dauphin Island. Oysters were tested for V. vulnificus and V. parahaemolyticus densities (by using the U.S. Food and Drug Administration enrichment method) after 2, 7, and 14 days deployed, with baseline samples taken (i) at the time of original harvest and iced, (ii) from oysters refrigerated within 1 h of harvest at oysters not refrigerated during the harvest trip (green tag) but refrigerated after an 8-h trip. White and green tag oysters were sampled ∼24 h on arrival in Bon Secour, AL, put on ice, and shipped for analysis. Among baseline samples, there were no significant differences in V. vulnificus and V. parahaemolyticus densities, although the densities in the green tag oysters tended to be highest. After transplanting, V. vulnificus densities were significantly highest on day 2, with no significant differences among any of the other days within a site. On day 2, Sandy Bay had significantly greater densities of V. vulnificus than the Dauphin Island site, but no other days differed from time zero. For Vibrio parahaemolyticus, densities were greatest on day 2 and lowest at time zero, but this did not differ significantly from abundance on day 14. Average survival was 83.4% (± 3.13 SD), with no differences between sites. These preliminary results indicate that high-salinity transplanting could be an effective method of converting green tag oysters to oysters suitable for

  5. Inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in oysters by high-hydrostatic pressure and mild heat.

    Science.gov (United States)

    Ye, Mu; Huang, Yaoxin; Chen, Haiqiang

    2012-10-01

    Several recent outbreaks associated with oysters have heightened safety concerns of raw shellfish consumptions, with the majority being attributed to Vibrio spp. The objective of this study was to determine the effect of high-hydrostatic pressure (HHP) followed by mild heating on the inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in live oysters. Inoculated oysters were randomly subjected to: a) pressurization at 200-300 MPa for 2 min at 21 °C, b) mild heat treatment at 40, 45 or 50 °C for up to 20 min and c) pressure treatment of 200-300 MPa for 2 min at 21 °C followed by heat treatment at 40-50 °C. Counts of V. parahaemolyticus and V. vulnificus were then determined using the most probable number (MPN) method. Pressurization at 200-300 MPa for 2 min resulted in various degrees of inactivation, from 1.2 to >7 log MPN/g reductions. Heat treatment at 40 and 45 °C for 20 min only reduced V. parahaemolyticus and V. vulnificus by 0.7-2.5 log MPN/g while at 50 °C for 15 min achieved >7 log MPN/g reduction. HHP and mild heat had synergistic effects. Combinations such as HHP at 250 MPa for 2 min followed by heat treatment at 45 °C for 15 min and HHP at 200 MPa for 2 min followed by heat treatment at 50 °C for 5 min reduced both V. parahaemolyticus and V. vulnificus to non-detectable levels by the MPN method (oysters (negative enrichment results). This study demonstrated the efficiency of HHP followed by mild heat treatments on inactivation of V. parahaemolyticus and V. vulnificus and could help the industry to establish parameters for processing oysters. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Vibrios patogênicos em ostras (Crassostrea rhizophorae servidas em restaurantes no Rio de Janeiro: um alerta para a Saúde Pública Pathogenic Vibrios in oysters (Crassostrea rhizophorae served at restaurants in Rio de Janeiro: a public health warning

    Directory of Open Access Journals (Sweden)

    Christiane Soares Pereira

    2007-06-01

    Full Text Available Avaliaram-se 40 amostras de ostras (Crassostrea rhizophorae servidas in natura em 15 restaurantes da Cidade do Rio de Janeiro, a fim de investigar a presença de Vibrio spp. As amostras de ostras foram analisadas e submetidas a enriquecimento em água peptonada alcalina adicionada de 1 e 3% de NaCl, incubadas a 37°C por 24 horas. Em seguida, os cultivos foram semeados em agar tiossulfato citrato bile sacarose e as colônias suspeitas foram submetidas à caracterização bioquímica. Vibrio parahaemolyticus, Vibrio carchariae, Vibrio alginolyticus e Vibrio vulnificus representaram as principais espécies (> 60% isoladas a partir das ostras in natura.Forty oyster samples (Crassostrea rhizophorae served raw in 15 restaurants in the city of Rio de Janeiro were evaluated in order to investigate the presence of Vibrio spp. The oyster samples were analyzed and subjected to enrichment in alkaline peptone water with the addition of 1 and 3% NaCl and incubated at 37°C for 24 hours. Following this, the cultures were seeded onto thiosulfate citrate bile sucrose agar (TCBS and the suspected colonies were subjected to biochemical characterization. Vibrio parahaemolyticus, Vibrio carchariae, Vibrio alginolyticus and Vibrio vulnificus were the main species (> 60% isolated from raw oysters.

  7. Extracellular proteolytic enzymes produced by human pathogenic Vibrio species

    Directory of Open Access Journals (Sweden)

    Shin-Ichi eMiyoshi

    2013-11-01

    Full Text Available Bacteria in the genus Vibrio produce extracellular proteolytic enzymes to obtain nutrients via digestion of various protein substrates. However, the enzymes secreted by human pathogenic species have been documented to modulate the bacterial virulence. Several species including Vibrio cholerae and V. vulnificus are known to produce thermolysin-like metalloproteases termed vibriolysin. The vibriolysin from V. vulnificus, a causative agent of serious systemic infection, is a major toxic factor eliciting the secondary skin damage characterized by formation of the hemorrhagic brae. The vibriolysin from intestinal pathogens may play indirect roles in pathogenicity because it can activate protein toxins and hemagglutinin by the limited proteolysis and can affect the bacterial attachment to or detachment from the intestinal surface by degradation of the mucus layer. Two species causing wound infections, V. alginolyticus and V. parahaemolyticus, produce another metalloproteases so-called collagenases. Although the detailed pathological roles have not been studied, the collagenase is potent to accelerate the bacterial dissemination through digestion of the protein components of the extracellular matrix. Some species produce cymotrypsin-like serine proteases, which may also affect the bacterial virulence potential. The intestinal pathogens produce sufficient amounts of the metalloprotease at the small intestinal temperature; however, the metalloprotease production by extra-intestinal pathogens is much higher around the body surface temperature. On the other hand, the serine protease is expressed only in the absence of the metalloprotease.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Aspects of vibrio parahaemolyticus(SAK) in fish preservation by irradiation 1. The presence of vibrio parahaemolyticus(SAK) in coastal areas of Sumatra and Java

    International Nuclear Information System (INIS)

    Suhadi, F.

    1984-01-01

    A study was carried out on the presence of vibrio parahaemolyticus in samples of sediment and seafoods originating from the eastern coast of Sumatra and the north coastal areas of Java. In a total of 2.434 samples of sediment and seafoods, 79 (3,3%) were found to contain vibrio parahaemolyticus. Among the 467 sediment samples, 1.650 fish, 133 shellfish, 123 shrimps, and 61 crab samples, 22 (4.7%), 41 (2.5%), 12 (9.0%), 2 (1,6%) and 2 (3.3%) were positive for vibrio parahaemolyticus, respectively. Based on the sampling areas, the high incidence of vibrio parahaemolyticus was found in samples collected from Riau (5.4%), while the samples from East Java was only about 0.4%. The contamination level of vibrio parahaemolyticus in samples collected from coastal areas of Sumatra and Java is relatively lower compared with the data obtained from some other coastal areas in the United States and Japan. (author)

  11. Structural organization of the transfer RNA operon I of Vibrio cholerae

    Indian Academy of Sciences (India)

    Unknown

    [Ghatak A, Majumdar A and Ghosh R K 2005 Structural organization of the transfer RNA operon I of Vibrio cholerae: Differences ..... clonal relationship are of utmost importance. ... rately derived from environmental, nontoxigenic, non-O1.

  12. Uptake of Vibrio cholerae biotype eltor from contaminated water by water hyacinth (eichornia crassipes).

    Science.gov (United States)

    Spira, W M; Huq, A; Ahmed, Q S; Saeed, Y A

    1981-09-01

    Vibrio cholerae biotype eltor appears to concentrate on the surface of the water hyacinth (Eichornia crassipes), thereby enhancing its survival and its potential for transmission through waterways of cholera-endemic regions such as Bangladesh.

  13. Uptake of Vibrio cholerae Biotype eltor from Contaminated Water by Water Hyacinth (Eichornia crassipes)

    OpenAIRE

    Spira, William M.; Huq, Anwarul; Ahmed, Qazi Shafi; Saeed, Yusuf A.

    1981-01-01

    Vibrio cholerae biotype eltor appears to concentrate on the surface of the water hyacinth (Eichornia crassipes), thereby enhancing its survival and its potential for transmission through waterways of cholera-endemic regions such as Bangladesh.

  14. Risk assessment of Vibrio parahaemolyticus in seafood: interpretative summary and technical report

    National Research Council Canada - National Science Library

    2011-01-01

    "Vibrio parahaemolyticus are common causes of diarrhoeal disease worldwide. These marine micro-organisms, native in estuarine waters globally, concentrate in the gut of filter-feeding molluscan shellfish, such as oysters, clams and mussels...

  15. INFLUENCE OF SEASONAL FACTORS ON OYSTER HEMOCYTE KILLING OF VIBRIO PARAHEMOLYTICUS

    Science.gov (United States)

    Seasonal variation of cellular defenses of oyster Crassostrea virginica against Vibrio parahaemolyticus were examined from June 1997 to December 1998 using a recently developed bactericidal assay that utilizes a tetrazolium dye. Mean hemocyte numbers, plasma lysozyme, and P. mari...

  16. DIFFERENTIAL EFFECTS OF OYSTER (CRASSOSTREA VIRGINICA) DEFENSES ON CLINICAL AND ENVIRONMENTAL ISOLATES OF VIBRIO PARAHEMOLYTICUS

    Science.gov (United States)

    Three clinical (2030, 2062, and 2107) and three environmental (1094, 1163, and ATCC 17802) isolates of Vibrio parahaemolyticus were exposed to hemocytes and plasma collected from oysters (Crassostrea virginica) to determine their susceptibility to putative oyster defenses. Clinic...

  17. Microbiota of Vibrio sp. in the hepatopancreas of cultured white pacific shrimp (Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Renata Albuquerque C.

    2013-08-01

    Full Text Available Objective. The present study aimed to investigate the presence of vibrios in the hepatopancreas of cultured shrimp. Materials and methods. Vibrios from the hepatopancreas of fifteen samples of five specimens each, of apparently healthy Pacific white shrimp (Litopenaeus vannamei were isolated, identified and quantified. Results. The vibrio density ranged from 430 to 2,400 MPN g-1 (rs MPN cm-1=-0.114; rs MPN g-1 = 0.211. Thirty isolations were obtained, most of which belonged to the species V. cholerae (n=11 and V. parahaemolyticus (n=7. Conclusions. The outcomes of the present study suggest that, even in the absence of symptoms of vibriosis, the microbiota of the hepatopancreas of cultured shrimp may include sucrose positive and negative vibrios.

  18. [Environmental drivers of emergence and spreading of Vibrio epidemics in South America].

    Science.gov (United States)

    Gavilán, Ronnie G; Martínez-Urtaza, Jaime

    2011-03-01

    Vibrio cholerae and V. parahaemolyticus are the two Vibrio species with a major impact on human health. Diseases caused by both pathogens are acquiring increasing relevance due to their expansion at global scale. In this paper, we resume the ecological aspects associated with the arrival and spreading of infections caused by V. parahaemolyticus and V. cholerae in Peru from a South American perspective. Moreover, we discuss the similarities in the emergence in Peru of cholera cases in 1991 and V. parahaemolyticus infections in 1997. These constituted exceptional experiments to evaluate the relationships between the Vibrio epidemics and changes in the environment. The epidemic radiations of V. cholerae and V. parahaemolyticus constitute to clear examples supporting the oceanic dispersion of pathogenic vibrios and have enabled the identification of El Niño events as a potential mechanism for the spreading of diseases through the ocean.

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

    Energy Technology Data Exchange (ETDEWEB)

    Colwell, Rita

    2012-06-01

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

  20. Antibiotic resistance monitoring in Vibrio spp. isolated from rearing environment and intestines of abalone Haliotis diversicolor.

    Science.gov (United States)

    Wang, R X; Wang, J Y; Sun, Y C; B L Yang; A L Wang

    2015-12-30

    546 Vibrio isolates from rearing seawater (292 strains) and intestines of abalone (254 strains) were tested to ten antibiotics using Kirby-Bauer diffusion method. Resistant rates of abalone-derived Vibrio isolates to chloramphenicol (C), enrofloxacin (ENX) and norfloxacin (NOR) were 40%) to kanamycin (KNA), furazolidone (F), tetracycline (TE), gentamicin (GM) and rifampin (RA). 332 isolates from seawater (n=258) and abalone (n=74) were resistant to more than three antibiotics. Peaked resistant rates of seawater-derived isolates to multiple antibiotics were overlapped in May and August. Statistical analysis showed that pH had an important effect on resistant rates of abalone-derived Vibrio isolates to RA, NOR, and ENX. Salinity and dissolved oxygen were negatively correlated with resistant rates of seawater-derived Vibrio isolates to KNA, RA, and PG. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Reduction of acute toxicity of the pharmaceutical fluoxetine (Prozac) submitted to ionizing radiation to Vibrio fischeri

    Energy Technology Data Exchange (ETDEWEB)

    Santos, Dymes R.A.; Garcia, Vanessa S.G.; Vilarrubia, Anna C.S.; Borrely, Sueli I., E-mail: vanessagarcia@usp.br, E-mail: sborrely@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2011-07-01

    The constant use of pharmaceutical drugs by great part of the population and its continuous input into the environment creates a growing need of investigating its presence, behavior and the effects on aquatic biota, as well as new ways to treat wastewater containing such substances. The fluoxetine hydrochloride (FH) present in the drug Prozac is an active ingredient used in the treatment of depressive and anxiety disorders. Generally, these compounds enter the aquatic environment by sewage collectors systems after undergoing prior treatment in sewage treatment plants (STPs) or without any treatment. This study focused on evaluating the reduction of acute toxicity of the pharmaceutical FH, under its manipulated formula, for the marine bacterium Vibrio fischeri. It was also evaluated the acute toxicity of the aqueous solution containing the FH after its exposition to ionizing radiation from industrial electron accelerator. It was performed acute toxicity tests lasting 15 minutes, where the average EC (50) of the non-irradiated CF water solution was approximately 0.68 mg L-1. While the CF water solution irradiated with 1 kGy, 2.5 kGy, 7.5 kGy and 10 kGy, presented an average EC(50) 1.63 mg.L{sup -1}, 2.34 mg.L{sup -1}, 2.35 mg.L{sup -1} and 1.80 mg.L{sup -1}, respectively, showing a notable reduction of the acute toxicity for this organism. (author)

  2. Mechanism of membrane damage by El Tor hemolysin of Vibrio cholerae O1.

    Science.gov (United States)

    Ikigai, H; Akatsuka, A; Tsujiyama, H; Nakae, T; Shimamura, T

    1996-08-01

    El Tor hemolysin (ETH; molecular mass, 65 kDa) derived from Vibrio cholerae O1 spontaneously assembled oligomeric aggregates on the membranes of rabbit erythrocyte ghosts and liposomes. Membrane-associated oligomers were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting into two to nine bands with apparent molecular masses of 170 to 350 kDa. ETH assembled oligomers on a liposomal membrane consisting of phosphatidylcholine and cholesterol, but not on a membrane of phosphatidylcholine alone. Cholesterol could be replaced with diosgenin or ergosterol but not with 5alpha-cholestane-3-one, suggesting that sterol is essential for the oligomerization. The treatment of carboxyfluorescein-encapsulated liposomes with ETH caused a rapid release of carboxyfluorescein into the medium. Because dextrin 20 (molecular mass, 900 Da) osmotically protected ETH-mediated hemolysis, this hemolysis is likely to be caused by pore formation on the membrane. The pore size(s) estimated from osmotic protection assays was in the range of 1.2 to 1.6 nm. The pore formed on a rabbit erythrocyte membrane was confirmed morphologically by electron microscopy. Thus, we provide evidence that ETH damages the target by the assembly of hemolysin oligomers and pore formation on the membrane.

  3. Reduction of acute toxicity of the pharmaceutical fluoxetine (Prozac) submitted to ionizing radiation to Vibrio fischeri

    International Nuclear Information System (INIS)

    Santos, Dymes R.A.; Garcia, Vanessa S.G.; Vilarrubia, Anna C.S.; Borrely, Sueli I.

    2011-01-01

    The constant use of pharmaceutical drugs by great part of the population and its continuous input into the environment creates a growing need of investigating its presence, behavior and the effects on aquatic biota, as well as new ways to treat wastewater containing such substances. The fluoxetine hydrochloride (FH) present in the drug Prozac is an active ingredient used in the treatment of depressive and anxiety disorders. Generally, these compounds enter the aquatic environment by sewage collectors systems after undergoing prior treatment in sewage treatment plants (STPs) or without any treatment. This study focused on evaluating the reduction of acute toxicity of the pharmaceutical FH, under its manipulated formula, for the marine bacterium Vibrio fischeri. It was also evaluated the acute toxicity of the aqueous solution containing the FH after its exposition to ionizing radiation from industrial electron accelerator. It was performed acute toxicity tests lasting 15 minutes, where the average EC (50) of the non-irradiated CF water solution was approximately 0.68 mg L-1. While the CF water solution irradiated with 1 kGy, 2.5 kGy, 7.5 kGy and 10 kGy, presented an average EC(50) 1.63 mg.L -1 , 2.34 mg.L -1 , 2.35 mg.L -1 and 1.80 mg.L -1 , respectively, showing a notable reduction of the acute toxicity for this organism. (author)

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

    Directory of Open Access Journals (Sweden)

    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.

  5. Induction of a gradual, reversible morphogenesis of its host's epithelial brush border by Vibrio fischeri.

    Science.gov (United States)

    Lamarcq, L H; McFall-Ngai, M J

    1998-02-01

    Bacteria exert a variety of influences on the morphology and physiology of animal cells whether they are pathogens or cooperative partners. The association between the luminous bacterium Vibrio fischeri and the sepiolid squid Euprymna scolopes provides an experimental model for the study of the influence of extracellular bacteria on the development of host epithelia. In this study, we analyzed bacterium-induced changes in the brush borders of the light organ crypt epithelia during the initial hours following colonization of this tissue. Transmission electron microscopy of the brush border morphology in colonized and uncolonized hosts revealed that the bacteria effect a fourfold increase in microvillar density over the first 4 days of the association. Estimates of the proportions of bacterial cells in contact with host microvilli showed that the intimacy of the bacterial cells with animal cell surfaces increases significantly during this time. Antibiotic curing of the organ following colonization showed that sustained interaction with bacteria is essential for the retention of the induced morphological changes. Bacteria that are defective in either light production or colonization efficiency produced changes similar to those by the parent strain. Conventional fluorescence and confocal scanning laser microscopy revealed that the brush border is supported by abundant filamentous actin. However, in situ hybridization with beta-actin probes did not show marked bacterium-induced increases in beta-actin gene expression. These experiments demonstrate that the E. scolopes-V. fischeri system is a viable model for the experimental study of bacterium-induced changes in host brush border morphology.

  6. Genome Sequence of Vibrio cholerae Strain O1 Ogawa El Tor, Isolated in Mexico, 2013.

    Science.gov (United States)

    Díaz-Quiñonez, José Alberto; Hernández-Monroy, Irma; López-Martínez, Irma; Ortiz-Alcántara, Joanna; González-Durán, Elizabeth; Ruiz-Matus, Cuitláhuac; Kuri-Morales, Pablo; Ramírez-González, José Ernesto

    2014-10-30

    We present the draft genome sequence of Vibrio cholerae InDRE 3140 recovered in 2013 during a cholera outbreak in Mexico. The genome showed the Vibrio 7th pandemic islands VSP1 and VSP2, the pathogenic islands VPI-1 and VPI-2, the integrative and conjugative element SXT/R391 (ICE-SXT), and both prophages CTXφ and RS1φ. Copyright © 2014 Díaz-Quiñonez et al.

  7. Prevalence and diversity of Aeromonas and Vibrio spp. in coastal waters of Southern Italy

    DEFF Research Database (Denmark)

    Dumontet, S.; Krovacek, K.; Svenson, S.B.

    2000-01-01

    % of samples were positive for Vibrio spp. It was interesting to note that 38% of the positive stations for both Aeromonas and Vibrio spp. showed a fecal coliform contamination of water at ... coliforms) do not always satisfactorily reflect the hygienic quality of water. The presence of Vibrionaceae on copepods was also investigated. Copepods were sampled at a station located inside the harbour of the city of Naples and were found contaminated by V. cholerae non-Ol, V. alginolyticus, V. fluvialis...

  8. 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.

  9. [Factors of persistence and (or) pathogenicity in vibrios and aeromonads belonging to different ecotopes].

    Science.gov (United States)

    Bukharin, O V; Boĭko, A V; Zhuravleva, L A

    1998-01-01

    Factors of persistence and/or pathogenicity in Vibrio parahaemolyticus and Aeromonas hydrophila (hemolytic, lipase, lecithin, DNAase, RNAase, antilysozyme, "anti-interferon", anticomplementary activities and capacity for absorbing Congo red) were studied. The study revealed the interspecific and subpopulation (hospital and extraorganismal parts of the population) differences in the activity of the manifestation of these factors. Strong dependence of the whole complex of persistence and pathogenicity factors of their belonging to the hostal part of Vibrio and Aeromonas populations was shown.

  10. Natural transformation of Vibrio parahaemolyticus: A rapid method to create genetic deletions.

    Science.gov (United States)

    Chimalapati, Suneeta; de Souza Santos, Marcela; Servage, Kelly; De Nisco, Nicole J; Dalia, Ankur B; Orth, Kim

    2018-03-19

    The Gram-negative bacterium Vibrio parahaemolyticus is an opportunistic human pathogen and the leading cause of seafood borne acute gastroenteritis worldwide. Recently, this bacterium was implicated as the etiologic agent of a severe shrimp disease with consequent devastating outcomes to shrimp farming. In both cases, acquisition of genetic material via horizontal transfer provided V. parahaemolyticus with new virulence tools to cause disease. Dissecting the molecular mechanisms of V. parahaemolyticus pathogenesis often requires manipulating its genome. Classically, genetic deletions in V. parahaemolyticus are performed using a laborious, lengthy, multi-step process. Herein, we describe a fast and efficient method to edit this bacterium's genome based on V. parahaemolyticus natural competence. Although this method is similar to one previously described, V. parahaemolyticus requires counter selection for curing of acquired plasmids due to its recalcitrant nature of retaining extrachromosomal DNA. We believe this approach will be of use to the Vibrio community. Importance Spreading of Vibrios throughout the world correlates with increased global temperatures. As they spread, they find new niches to survive, proliferate and invade. Therefore, genetic manipulation of Vibrios is of utmost importance for studying these species. Herein, we have delineated and validated a rapid method to create genetic deletions in Vibrio parahaemolyticus This study provides insightful methodology for studies with other Vibrio species. Copyright © 2018 American Society for Microbiology.

  11. Prevalence of potentially pathogenic Vibrio species in the seafood marketed in Malaysia.

    Science.gov (United States)

    Elhadi, Nasreldin; Radu, Son; Chen, Chien-Hsien; Nishibuchi, Mitsuaki

    2004-07-01

    Seafood samples obtained in seafood markets and supermarkets at 11 sites selected from four states in Malaysia were examined for the presence of nine potentially pathogenic species from the genus Vibrio between July 1998 and June 1999. We examined 768 sample sets that included shrimp, squid, crab, cockles, and mussels. We extensively examined shrimp samples from Selangor State to determine seasonal variation of Vibrio populations. Eight potentially pathogenic Vibrio species were detected, with overall incidence in the samples at 4.6% for V. cholerae, 4.7% for V. parahaemolyticus, 6.0% for V. vulnificus, 11% for V. alginolyticus, 9.9% for V. metschnikovii, 1.3% for V. mimicus, 13% for V. damsela, 7.6% for V. fluvialis, and 52% for a combined population of all of the above. As many as eight Vibrio species were detected in shrimp and only four in squid and peel mussels. The overall percent incidence of any of the eight vibrios was highest (82%) in cockles (Anadara granosa) among the seafoods examined and was highest (100%) in Kuching, Sarawak State, and lowest (25%) in Penang, Pulau Penang State, among the sampling sites. Of 97 strains of V. cholerae isolated, one strain belonged to the O1 serotype and 14 to the O139 serotype. The results indicate that the various seafood markets in Malaysia are contaminated with potentially pathogenic Vibrio species regardless of the season and suggest that there is a need for adequate consumer protection measures.

  12. Prevalence of listeria, Aeromonas, and Vibrio species in fish used for human consumption in Turkey.

    Science.gov (United States)

    Yücel, Nihal; Balci, Senay

    2010-02-01

    A total of 78 raw retail fish samples from 30 freshwater and 48 marine fish were examined for the presence of Listeria, Aeromonas, and Vibrio species. The overall incidence of Listeria spp. was 30% in freshwater samples and 10.4% in marine fish samples. Listeria monocytogenes (44.5%) was the most commonly isolated species in freshwater fish, and Listeria murrayi (83.5%) was the most commonly isolated species in marine fish samples. Motile aeromonads were more common in marine fish samples (93.7%) than in freshwater fish samples (10%). Vibrio alginolyticus, Vibrio fluvialis, and Vibrio damsela were isolated only in marine fish samples, representing 40.9, 38.6, and 36.3% of Vibrio isolates, respectively. In freshwater and marine fish, the highest incidences of Listeria and Aeromonas were found in skin samples; the highest incidence of Vibrio in marine fish was found in gill samples. The location of Listeria spp. and L. monocytogenes in a fish was significantly different among freshwater fish. A high incidence of these bacterial pathogens was found in the brown trout (Salmo trutta) and horse mackerel (Trachurus trachurus). Handling of contaminated fish, cross-contamination, or eating raw fish might pose a health hazard, especially in immunosuppressed individuals, elderly people, and children. This study highlights the importance of bacterial pathogens in fish intended for human consumption, but more study is needed.

  13. Spatiotemporal Dynamics of Total Viable Vibrio spp. in a NW Mediterranean Coastal Area.

    Science.gov (United States)

    Girard, Léa; Peuchet, Sébastien; Servais, Pierre; Henry, Annabelle; Charni-Ben-Tabassi, Nadine; Baudart, Julia

    2017-09-27

    A cellular approach combining Direct Viable Counting and Fluorescent In Situ Hybridization using a one-step multiple-probe technique and Solid Phase Cytometry (DVC-FISH-SPC) was developed to monitor total viable vibrios and cover the detection of a large diversity of vibrios. FISH combined three probes in the same assay and targeted sequences located at different positions on the 16S rRNA of Vibrio and Aliivibrio members. We performed a 10-month in situ study to investigate the weekly dynamics of viable vibrios relative to culturable counts at two northwestern Mediterranean coastal sites, and identified the key physicochemical factors for their occurrence in water using a multivariate analysis. Total viable and culturable cell counts showed the same temporal pattern during the warmer season, whereas the ratios between both methods were inverted during the colder seasons (<15°C), indicating that some of the vibrio community had entered into a viable but non-culturable (VBNC) state. We confirmed that Seawater Surface Temperature explained 51-62% of the total variance in culturable counts, and also showed that the occurrence of viable vibrios is controlled by two variables, pheopigment (15%) and phosphate (12%) concentrations, suggesting that other unidentified factors play a role in maintaining viability.

  14. Acute Otitis due to Vibrio fluvialis after Swimming

    Directory of Open Access Journals (Sweden)

    Ping-Jen Chen

    2012-01-01

    Full Text Available A 40-year-old female presented with purulent exudate through the left auditive duct and pain in the left ear region, which intensified during mastication. After collection of the pus from the left ear lesion, amoxicillin-clavulanic acid for seven days was prescribed for a presumed diagnosis of acute otitis. Four days later, the pus culture grew V. fluvialis which is further identified by API 20E identification system (bioMérieux. Following the successful completion of a course of antibiotics, the patient recovered completely and without complication. To the best of our knowledge, this is the first case of Vibrio fluvialis otitis after swimming in an immunocompetent patient.

  15. Aptamer-Based Paper Strip Sensor for Detecting Vibrio fischeri.

    Science.gov (United States)

    Shin, Woo-Ri; Sekhon, Simranjeet Singh; Rhee, Sung-Keun; Ko, Jung Ho; Ahn, Ji-Young; Min, Jiho; Kim, Yang-Hoon

    2018-05-14

    Aptamer-based paper strip sensor for detecting Vibrio fischeri was developed. Our method was based on the aptamer sandwich assay between whole live cells, V. fischeri and DNA aptamer probes. Following 9 rounds of Cell-SELEX and one of the negative-SELEX, V. fischeri Cell Aptamer (VFCA)-02 and -03 were isolated, with the former showing approximately 10-fold greater avidity (in the subnanomolar range) for the target cells when arrayed on a surface. The colorimetric response of a paper sensor based on VFCA-02 was linear in the range of 4 × 10 1 to 4 × 10 5 CFU/mL of target cell by using scanning reader. The linear regression correlation coefficient ( R 2 ) was 0.9809. This system shows promise for use in aptamer-conjugated gold nanoparticle probes in paper strip format for in-field detection of marine bioindicating bacteria.

  16. In situ measured elimination of Vibrio cholerae from brackish water.

    Science.gov (United States)

    Pérez, María Elena Martínez; Macek, Miroslav; Galván, María Teresa Castro

    2004-01-01

    In situ elimination of fluorescently labelled Vibrio cholerae (FLB) was measured in two saline water bodies in Mexico: in a brackish water lagoon, Mecoacán (Gulf of Mexico; State of Tabasco) and an athalassohaline lake, Alchichica (State of Puebla). Disappearance rates of fluorescently labelled V. cholera O1 showed that they were eliminated from the environment at an average rate of 32% and 63%/day, respectively (based on the bacterial standing stocks). The indirect immunofluorescence method confirmed the presence of V. cholerae O1 in the lagoon. However, the elimination of FLB was not directly related either to the presence or absence of the bacterium in the water body or to the phytoplankton concentration.

  17. Attenuation of Vibrio fischeri quorum sensing using rationally designed polymers.

    Science.gov (United States)

    Piletska, Elena V; Stavroulakis, Georgios; Karim, Kal; Whitcombe, Michael J; Chianella, Iva; Sharma, Anant; Eboigbodin, Kevin E; Robinson, Gary K; Piletsky, Sergey A

    2010-04-12

    A first attempt to attenuate the quorum sensing (QS) of a marine heterotroph microorganism, Vibrio fischeri , using signal molecule-sequestering polymers (SSPs) is presented. A set of rationally designed polymers with affinity toward a signal molecule of V. fischeri , N-(beta-ketocaproyl)-l-homoserine lactone (3-oxo-C6-AHL) was produced. It is reported that computationally designed polymers could sequester a signal molecule of V. fischeri and prevent QS-controlled phenotypes (in this case, bioluminescence) from being up-regulated. It was proven that the attenuation of bioluminescence of V. fischeri was due to sequestration of the signal molecule by specific polymers and not due to the toxicity of polymer or nonspecific depletion of nutrients. The ability to disrupt the bacterial communication using easy to synthesize and chemically inert polymers could provide a new concept for the development of pharmaceuticals and susceptible device coatings such as catheters.

  18. Vibrio cholerae as a predator: lessons from evolutionary principles

    Directory of Open Access Journals (Sweden)

    Stefan ePukatzki

    2013-12-01

    Full Text Available Diarrheal diseases are the second-most common cause of death among children under the age of five worldwide. Cholera alone, caused by the marine bacterium Vibrio cholerae, is responsible for several million cases and over 120,000 deaths annually. When contaminated water is ingested, V. cholerae passes through the gastric acid barrier, penetrates the mucin layer of the small intestine, and adheres to the underlying epithelial lining. V. cholerae multiplies rapidly, secretes cholera toxin, and exits the human host in vast numbers during diarrheal purges. How V. cholerae rapidly reaches such high numbers during each purge is not clearly understood. We propose that V. cholerae employs its bactericidal type VI secretion system to engage in intraspecies and intraguild predation for nutrient acquisition to support rapid growth and multiplication.

  19. Impacts of Climatic Variability on Vibrio parahaemolyticus Outbreaks in Taiwan

    Science.gov (United States)

    Hsiao, Hsin-I; Jan, Man-Ser; Chi, Hui-Ju

    2016-01-01

    This study aimed to investigate and quantify the relationship between climate variation and incidence of Vibrio parahaemolyticus in Taiwan. Specifically, seasonal autoregressive integrated moving average (ARIMA) models (including autoregression, seasonality, and a lag-time effect) were employed to predict the role of climatic factors (including temperature, rainfall, relative humidity, ocean temperature and ocean salinity) on the incidence of V. parahaemolyticus in Taiwan between 2000 and 2011. The results indicated that average temperature (+), ocean temperature (+), ocean salinity of 6 months ago (+), maximum daily rainfall (current (−) and one month ago (−)), and average relative humidity (current and 9 months ago (−)) had significant impacts on the incidence of V. parahaemolyticus. Our findings offer a novel view of the quantitative relationship between climate change and food poisoning by V. parahaemolyticus in Taiwan. An early warning system based on climate change information for the disease control management is required in future. PMID:26848675

  20. Bile Sensing: The Activation of Vibrio parahaemolyticus Virulence

    Directory of Open Access Journals (Sweden)

    Bey-Hing Goh

    2017-04-01

    Full Text Available Bacteria must develop resistance to various inhospitable conditions in order to survive in the human gastrointestinal tract. Bile, which is secreted by the liver, and plays an important role in food digestion also has antimicrobial properties and is able to disrupt cellular homeostasis. Paradoxically, although bile is one of the guts defenses, many studies have reported that bacteria such as Vibrio parahaemolyticus can sense bile and use its presence as an environmental cue to upregulate virulence genes during infection. This article aims to discuss how bile is detected by V. parahaemolyticus and its role in regulating type III secretion system 2 leading to human infection. This bile–bacteria interaction pathway gives us a clearer understanding of the biochemical and structural analysis of the bacterial receptors involved in mediating a response to bile salts which appear to be a significant environmental cue during initiation of an infection.

  1. Impacts of Climatic Variability on Vibrio parahaemolyticus Outbreaks in Taiwan

    Directory of Open Access Journals (Sweden)

    Hsin-I Hsiao

    2016-02-01

    Full Text Available This study aimed to investigate and quantify the relationship between climate variation and incidence of Vibrio parahaemolyticus in Taiwan. Specifically, seasonal autoregressive integrated moving average (ARIMA models (including autoregression, seasonality, and a lag-time effect were employed to predict the role of climatic factors (including temperature, rainfall, relative humidity, ocean temperature and ocean salinity on the incidence of V. parahaemolyticus in Taiwan between 2000 and 2011. The results indicated that average temperature (+, ocean temperature (+, ocean salinity of 6 months ago (+, maximum daily rainfall (current (− and one month ago (−, and average relative humidity (current and 9 months ago (− had significant impacts on the incidence of V. parahaemolyticus. Our findings offer a novel view of the quantitative relationship between climate change and food poisoning by V. parahaemolyticus in Taiwan. An early warning system based on climate change information for the disease control management is required in future.

  2. Impacts of Climatic Variability on Vibrio parahaemolyticus Outbreaks in Taiwan.

    Science.gov (United States)

    Hsiao, Hsin-I; Jan, Man-Ser; Chi, Hui-Ju

    2016-02-03

    This study aimed to investigate and quantify the relationship between climate variation and incidence of Vibrio parahaemolyticus in Taiwan. Specifically, seasonal autoregressive integrated moving average (ARIMA) models (including autoregression, seasonality, and a lag-time effect) were employed to predict the role of climatic factors (including temperature, rainfall, relative humidity, ocean temperature and ocean salinity) on the incidence of V. parahaemolyticus in Taiwan between 2000 and 2011. The results indicated that average temperature (+), ocean temperature (+), ocean salinity of 6 months ago (+), maximum daily rainfall (current (-) and one month ago (-)), and average relative humidity (current and 9 months ago (-)) had significant impacts on the incidence of V. parahaemolyticus. Our findings offer a novel view of the quantitative relationship between climate change and food poisoning by V. parahaemolyticus in Taiwan. An early warning system based on climate change information for the disease control management is required in future.

  3. Vibrio cholerae infection, novel drug targets and phage therapy.

    Science.gov (United States)

    Fazil, Mobashar Hussain Urf Turabe; Singh, Durg V

    2011-10-01

    Vibrio cholerae is the causative agent of the diarrheal disease cholera. Although antibiotic therapy shortens the duration of diarrhea, excessive use has contributed to the emergence of antibiotic resistance in V. cholerae. Mobile genetic elements have been shown to be largely responsible for the shift of drug resistance genes in bacteria, including some V. cholerae strains. Quorum sensing communication systems are used for interaction among bacteria and for sensing environmental signals. Sequence analysis of the ctxB gene of toxigenic V. cholerae strains demonstrated its presence in multiple cholera toxin genotypes. Moreover, bacteriophage that lyse the bacterium have been reported to modulate epidemics by decreasing the required infectious dose of the bacterium. In this article, we will briefly discuss the disease, its clinical manifestation, antimicrobial resistance and the novel approaches to locate drug targets to treat cholera.

  4. Sequence and features of the tryptophan operon of Vibrio parahemolyticus.

    Science.gov (United States)

    Crawford, I P; Han, C Y; Silverman, M

    1991-01-01

    The nucleotide sequence of the trp operon of the marine enteric bacterium Vibrio parahemolyticus is presented. The gene order E, G, D, C(F), B, A is identical to that of other enterics. The structural genes of the operon are preceded by a long leader region encoding a 41-residue peptide containing five tryptophan residues. The organization of the leader region suggests that transcription of the operon is subject to attenuation control. The promoter-operator region of the V. parahemolyticus trp operon is almost identical to the corresponding promoter-operator of E. coli. The similarities suggest that promoter strength and operator function are identical in the two species, and that transcription initiation is regulated by repression. The operon appears to lack the internal promoter within trpD that is common in terrestrial enteric species.

  5. Effects of Dry Storage and Resubmersion of Oysters on Total Vibrio vulnificus and Total and Pathogenic (tdh+/trh+) Vibrio parahaemolyticus Levels.

    Science.gov (United States)

    Kinsey, Thomas P; Lydon, Keri A; Bowers, John C; Jones, Jessica L

    2015-08-01

    Vibrio vulnificus (Vv) and Vibrio parahaemolyticus (Vp) are the two leading causes of bacterial illnesses associated with raw shellfish consumption. Levels of these pathogens in oysters can increase during routine antifouling aquaculture practices involving dry storage in ambient air conditions. After storage, common practice is to resubmerge these stored oysters to reduce elevated Vv and Vp levels, but evidence proving the effectiveness of this practice is lacking. This study examined the changes in Vv and in total and pathogenic (thermostable direct hemolysin gene and the tdh-related hemolysin gene, tdh+ and trh+) Vp levels in oysters after 5 or 24 h of dry storage (28 to 32°C), followed by resubmersion (27 to 32°C) for 14 days. For each trial, replicate oyster samples were collected at initial harvest, after dry storage, after 7 days, and after 14 days of resubmersion. Oysters not subjected to dry storage were collected and analyzed to determine natural undisturbed vibrio levels (background control). Vibrio levels were measured using a most-probable-number enrichment followed by real-time PCR. After storage, vibrio levels (excluding tdh+ and trh+ Vp during 5-h storage) increased significantly (P oysters stored for 5 h) were not significantly different (P oysters. Vv and total and pathogenic Vp levels were not significantly different (P > 0.1) from levels in background oysters after 14 days of resubmersion, regardless of dry storage time. These data demonstrate that oyster resubmersion after dry storage at elevated ambient temperatures allows vibrio levels to return to those of background control samples. These results can be used to help minimize the risk of Vv and Vp illnesses and to inform the oyster industry on the effectiveness of routine storing and resubmerging of aquaculture oysters.

  6. Ozone Disinfection of Vibrio vulnificus in Shrimp Pond Water

    Science.gov (United States)

    Dyah Pita Rengga, Wara; Cahya Julyta Putri, Echa; Wulansarie, Ria; Suryanto, Agus

    2018-03-01

    One variety of shrimp, L.Vanamei, often uses brackish water during the operation in the shrimp pond. Chlorination and ultraviolet are usually used for disinfection of brackish water. However, it is ineffective and forms sediment in the water distribution. It can be a negative impact on the water quality cause a contamination on the shrimp, so the farmers might have loss of profit because Vibrio vulnificus causes infection and dead on the shrimp. It affects the safety of consumers and should be minimized. The purpose of this study is to reduce the number of V. vulnificus bacteria in the pond water. The water was put in the storage tanks then pumped to filter out the impurities of the water. Furthermore, the water set the flow rate in 1 LPM, 2 LPM, and 3 LPM. After that, the ozone was injected to the water flow to sterilize the V. vulnificus bacteria. Finally, the water was returned to the original tank. The water from the tank was taken through a valve and analyzed in 0, 3, 7, 12, 18, 24, 30 minutes. The sample was analyzed immediately using a Total Plate Count method to determine the number of V. vulnificus bacteria in the shrimp pond water. The flow rate shows that the longer time of ozone made a lower amount of Vibrio v. bacteria. In 2 LPM water, it shows the optimum results of V. vulnificus. bacteria reduction for 88.1% compared to the flow rate of 1 LPM and 3 LPM with the bacteria reduction of 68,8% and 70.6%. This study shows that the ozone with a flow rate of 2 LPM circulation is the most effective method to help reducing the number of V. vulnificus in brackish water distribution system in the shrimp environment and potentially as a disinfectant.

  7. Dynamics of Vibrio with virulence genes detected in Pacific harbor seals (Phoca vitulina richardii) off California: implications for marine mammal health.

    Science.gov (United States)

    Hughes, Stephanie N; Greig, Denise J; Miller, Woutrina A; Byrne, Barbara A; Gulland, Frances M D; Harvey, James T

    2013-05-01

    Given their coastal site fidelity and opportunistic foraging behavior, harbor seals (Phoca vitulina) may serve as sentinels for coastal ecosystem health. Seals using urbanized coastal habitat can acquire enteric bacteria, including Vibrio that may affect their health. To understand Vibrio dynamics in seals, demographic and environmental factors were tested for predicting potentially virulent Vibrio in free-ranging and stranded Pacific harbor seals (Phoca vitulina richardii) off California. Vibrio prevalence did not vary with season and was greater in free-ranging seals (29 %, n = 319) compared with stranded seals (17 %, n = 189). Of the factors tested, location, turbidity, and/or salinity best predicted Vibrio prevalence in free-ranging seals. The relationship of environmental factors with Vibrio prevalence differed by location and may be related to oceanographic or terrestrial contributions to water quality. Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio cholerae were observed in seals, with V. cholerae found almost exclusively in stranded pups and yearlings. Additionally, virulence genes (trh and tdh) were detected in V. parahaemolyticus isolates. Vibrio cholerae isolates lacked targeted virulence genes, but were hemolytic. Three out of four stranded pups with V. parahaemolyticus (trh+ and/or tdh+) died in rehabilitation, but the role of Vibrio in causing mortality is unclear, and Vibrio expression of virulence genes should be investigated. Considering that humans share the environment and food resources with seals, potentially virulent Vibrio observed in seals also may be of concern to human health.

  8. Investigating the decay rates of Escherichia coli relative to Vibrio parahemolyticus and Salmonella Typhi in tropical coastal waters.

    Science.gov (United States)

    Lee, Choon Weng; Ng, Angie Yee Fang; Bong, Chui Wei; Narayanan, Kumaran; Sim, Edmund Ui Hang; Ng, Ching Ching

    2011-02-01

    Using the size fractionation method, we measured the decay rates of Escherichia coli, Salmonella Typhi and Vibrio parahaemolyticus in the coastal waters of Peninsular Malaysia. The size fractions were total or unfiltered, 0.7 μm) than in the smaller fraction (Vibrio grew well in seawater. There was usually an increase in Vibrio after one day incubation. Our results confirmed that decay or loss rates of E. coli did not match that of Vibrio, and also did not correlate with Salmonella decay rates. However E. coli showed persistence where its decay rates were generally lower than Salmonella. © 2010 Elsevier Ltd. All rights reserved.

  9. 养殖大黄鱼病原弧菌多重PCR检测技术的建立和应用%Development and application of multiplex PCR of vibrios pathogenic to cultured large yellow croaker, Pseudnosciaena crocea (Richardson)

    Institute of Scientific and Technical Information of China (English)

    祝璟琳; 王国良; 金珊

    2009-01-01

    溶藻弧菌(Vibrio alginolyticus)、副溶血弧菌(vibrio parahaemolyticus)和哈维氏弧菌(Vibrio harveyi)是浙江省养殖大黄鱼(Pseudnosciaena crocea)弧菌病的主要致病菌.本研究选择针对溶藻弧菌和副溶血弧菌的胶原酶基因,哈维氏弧菌的部分ToxR基因的特异性,优化设计了3对特异性引物,通过进行多重PCR反应体系优化,多重PCR产物的测序鉴定与特异性和敏感性实验,建立了一种检测致病性弧菌的多重PCR检测方法.经过琼脂糖凝胶电泳后的条带分析判断,可以在一个PCR管中同时成功地检测这3种病原细菌,含溶藻弧菌、哈维氏弧菌和副溶血弧菌3种致病弧菌核酸的阳性对照样品分别扩增出大小为737 hp、382 bp和271 bp的预期产物,其灵敏度是102~102 CFU/mL.将该方法应用于检测人工感染后的养殖大黄鱼病鱼肝脏和肾脏,结果在6份组织样本中,5份检出原始感染菌株,与API 20E鉴定结果相符;对弧菌病流行季节采集的未发病的16份养殖大黄鱼组织样本和16份水体样本进行抽检,结果在1份大黄鱼组织样本中检出哈维氏弧菌,7份水体样本中检出这3种弧菌中的1种或2种,鉴定结果与API 20E鉴定结果符合率为93.75%.说明该方法不仅可以检测发病鱼,还可以检测无病症带菌大黄鱼以及带菌水样,且说明海洋水体中存在着大黄鱼弧菌病的致病菌.结果说明,多重PCR检测方法具有较高的敏感性与特异性,可以缩短检测时间,降低检测成本,该方法的建立对养殖大黄鱼弧菌病的快速诊断和分子流行病学的调查具有重要意义.

  10. Recovery and evolutionary analysis of complete integron gene cassette arrays from Vibrio

    Directory of Open Access Journals (Sweden)

    Gillings Michael R

    2006-01-01

    Full Text Available Abstract Background Integrons are genetic elements capable of the acquisition, rearrangement and expression of genes contained in gene cassettes. Gene cassettes generally consist of a promoterless gene associated with a recombination site known as a 59-base element (59-be. Multiple insertion events can lead to the assembly of large integron-associated cassette arrays. The most striking examples are found in Vibrio, where such cassette arrays are widespread and can range from 30 kb to 150 kb. Besides those found in completely sequenced genomes, no such array has yet been recovered in its entirety. We describe an approach to systematically isolate, sequence and annotate large integron gene cassette arrays from bacterial strains. Results The complete Vibrio sp. DAT722 integron cassette array was determined through the streamlined approach described here. To place it in an evolutionary context, we compare the DAT722 array to known vibrio arrays and performed phylogenetic analyses for all of its components (integrase, 59-be sites, gene cassette encoded genes. It differs extensively in terms of genomic context as well as gene cassette content and organization. The phylogenetic tree of the 59-be sites collectively found in the Vibrio gene cassette pool suggests frequent transfer of cassettes within and between Vibrio species, with slower transfer rates between more phylogenetically distant relatives. We also identify multiple cases where non-integron chromosomal genes seem to have been assembled into gene cassettes and others where cassettes have been inserted into chromosomal locations outside integrons. Conclusion Our systematic approach greatly facilitates the isolation and annotation of large integrons gene cassette arrays. Comparative analysis of the Vibrio sp. DAT722 integron obtained through this approach to those found in other vibrios confirms the role of this genetic element in promoting lateral gene transfer and suggests a high rate of gene

  11. Relative contributions of Vibrio polysaccharide and quorum sensing to the resistance of Vibrio cholerae to predation by heterotrophic protists.

    Directory of Open Access Journals (Sweden)

    Shuyang Sun

    Full Text Available Protozoan grazing is a major mortality factor faced by bacteria in the environment. Vibrio cholerae, the causative agent of the disease cholera, is a natural inhabitant of aquatic ecosystems, and its survival depends on its ability to respond to stresses, such as predation by heterotrophic protists. Previous results show that grazing pressure induces biofilm formation and enhances a smooth to rugose morphotypic shift, due to increased expression of Vibrio polysaccharide (VPS. In addition to negatively controlling vps genes, the global quorum sensing (QS regulator, HapR, plays a role in grazing resistance as the ΔhapR strain is efficiently consumed while the wild type (WT is not. Here, the relative and combined contributions of VPS and QS to grazing resistance were investigated by exposing VPS and HapR mutants and double mutants in VPS and HapR encoding genes at different phases of biofilm development to amoeboid and flagellate grazers. Data show that the WT biofilms were grazing resistant, the VPS mutants were less resistant than the WT strain, but more resistant than the QS mutant strain, and that QS contributes to grazing resistance mainly in mature biofilms. In addition, grazing effects on biofilms of mixed WT and QS mutant strains were investigated. The competitive fitness of each strain in mixed biofilms was determined by CFU and microscopy. Data show that protozoa selectively grazed the QS mutant in mixed biofilms, resulting in changes in the composition of the mixed community. A small proportion of QS mutant cells which comprised 4% of the mixed biofilm biovolume were embedded in grazing resistant WT microcolonies and shielded from predation, indicating the existence of associational protection in mixed biofilms.

  12. 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.

    2010-01-01

    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. PMID:20817802

  13. 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.

  14. Ecology of Vibrio vulnificus in estuarine waters of eastern North Carolina.

    Science.gov (United States)

    Pfeffer, Courtney S; Hite, M Frances; Oliver, James D

    2003-06-01

    While several studies on the ecology of Vibrio vulnificus in Gulf Coast environments have been reported, there is little information on the distribution of this pathogen in East Coast waters. Thus, we conducted a multiyear study on the ecology of V. vulnificus in estuarine waters of the eastern United States, employing extensive multiple regression analyses to reveal the major environmental factors controlling the presence of this pathogen, and of Vibrio spp., in these environments. Monthly field samplings were conducted between July 2000 and April 2002 at six different estuarine sites along the eastern coast of North Carolina. At each site, water samples were taken and nine physicochemical parameters were measured. V. vulnificus isolates, along with estuarine bacteria, Vibrio spp., Escherichia coli organisms, and total coliforms, were enumerated in samples from each site by using selective media. During the last 6 months of the study, sediment samples were also analyzed for the presence of vibrios, including V. vulnificus. Isolates were confirmed as V. vulnificus by using hemolysin gene PCR or colony hybridization. V. vulnificus was isolated only when water temperatures were between 15 and 27 degrees C, and its presence correlated with water temperature and dissolved oxygen and vibrio levels. Levels of V. vulnificus in sediments were low, and no evidence for an overwintering in this environment was found. Multiple regression analysis indicated that vibrio levels were controlled primarily by temperature, turbidity, and levels of dissolved oxygen, estuarine bacteria, and coliforms. Water temperature accounted for most of the variability in the concentrations of both V. vulnificus (47%) and Vibrio spp. (48%).

  15. 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.

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

    Directory of Open Access Journals (Sweden)

    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

  17. An outbreak of Vibrio cholerae in Vikas Nagar, Chandigarh, India

    Directory of Open Access Journals (Sweden)

    Sonia Puri

    2014-01-01

    Full Text Available Background : On 1 July 2012, a large number of cases of acute diarrheal episodes were reported in Vikas Nagar, Chandigarh. A rapid response team was sent to investigate this outbreak on 3 July 2012. Aim : To determine the reasons for the outbreak and to focus on the identification of a gap in the management of the epidemic by applying remedial measures in the Vibrio cholera outbreak in the Vikas Nagar area of Chandigarh district. Materials and Methods : A house-to-house survey of 2765 houses was performed with 20 teams of Auxillary Nurse Midwife ANM/Anganwadi workers. Information regarding age, sex, place of residence, occupation, date of onset and treatment history and laboratory finding were collected. Environmental investigation and laboratory investigation of the stool samples were also performed. As the study was conducted during an emergency response to the outbreak, and was designed to provide information to orient the public health response, ethical approval was not required. Remedial measures were implemented. Results : A total of 1875 patients reported to the various health facilities of the Vikas Nagar area with complaints of increased frequency of loose watery diarrhea and a few had vomiting episodes during the time period of 1 - 14 July 2012. Four deaths were reported. Three hundred eighteen (318 cases were found in the house-to-house survey of 2765 houses of the area. Twenty-six percent of the cases were in the age group of 1800 MPN/100 mL was reported from 10 water samples. Investigations revealed that the epidemic was waterborne. Leakages in the pipes were found at many places leading to mixing of water with drainage, and water samples collected from the houses of the cases were found to be positive for Vibrio cholerae. Conclusion : Among the identified gaps, delays in the initiation of the investigation of the epidemic and pipe leakages were the most important. In India, waterborne epidemics are usual occurrences during the year

  18. Vibrio cholerae Colonization of Soft-Shelled Turtles.

    Science.gov (United States)

    Wang, Jiazheng; Yan, Meiying; Gao, He; Lu, Xin; Kan, Biao

    2017-07-15

    Vibrio cholerae is an important human pathogen and environmental microflora species that can both propagate in the human intestine and proliferate in zooplankton and aquatic organisms. Cholera is transmitted through food and water. In recent years, outbreaks caused by V. cholerae -contaminated soft-shelled turtles, contaminated mainly with toxigenic serogroup O139, have been frequently reported, posing a new foodborne disease public health problem. In this study, the colonization by toxigenic V. cholerae on the body surfaces and intestines of soft-shelled turtles was explored. Preferred colonization sites on the turtle body surfaces, mainly the carapace and calipash of the dorsal side, were observed for the O139 and O1 strains. Intestinal colonization was also found. The colonization factors of V. cholerae played different roles in the colonization of the soft-shelled turtle's body surface and intestine. Mannose-sensitive hemagglutinin (MSHA) of V. cholerae was necessary for body surface colonization, but no roles were found for toxin-coregulated pili (TCP) or N -acetylglucosamine-binding protein A (GBPA). Both TCP and GBPA play important roles for colonization in the intestine, whereas the deletion of MSHA revealed only a minor colonization-promoting role for this factor. Our study demonstrated that V. cholerae can colonize the surfaces and the intestines of soft-shelled turtles and indicated that the soft-shelled turtles played a role in the transmission of cholera. In addition, this study showed that the soft-shelled turtle has potential value as an animal model in studies of the colonization and environmental adaption mechanisms of V. cholerae in aquatic organisms. IMPORTANCE Cholera is transmitted through water and food. Soft-shelled turtles contaminated with Vibrio cholerae (commonly the serogroup O139 strains) have caused many foodborne infections and outbreaks in recent years, and they have become a foodborne disease problem. Except for epidemiological

  19. 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

  20. 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.

  1. Increases in the amounts of Vibrio spp. in oysters upon addition of exogenous bacteria.

    Science.gov (United States)

    Froelich, Brett; Oliver, James

    2013-09-01

    The bacterial pathogen Vibrio vulnificus is found naturally in brackish coastal waters but can be greatly concentrated by filter-feeding organisms such as shellfish. Numerous experiments in which exogenous V. vulnificus cells are added to oysters in an attempt to measure uptake and depuration have been performed. In nearly all cases, results have shown that laboratory-grown bacteria are rapidly taken up by the oysters but ultimately eliminated, while naturally present Vibrio populations in oysters are resistant to depuration. In this study, oysters harvested during winter months, with low culturable Vibrio concentrations, were incubated in aquaria supplemented with strains of V. vulnificus that were either genotypically or phenotypically distinct from the background bacteria. These exogenous cells were eliminated from the oysters, as previously seen, but other vibrios already inhabiting the oysters responded to the V. vulnificus inoculum by rapidly increasing in number and maintaining a large stable population. The presence of such an oyster-adapted Vibrio population would be expected to prevent colonization by exogenous V. vulnificus cells, thus explaining the rapid depuration of these added bacteria.

  2. Occurrence of Vibrio and Salmonella species in mussels (Mytilus galloprovincialis) collected along the Moroccan Atlantic coast.

    Science.gov (United States)

    Mannas, Hasna; Mimouni, Rachida; Chaouqy, Noureddine; Hamadi, Fatima; Martinez-Urtaza, Jaime

    2014-01-01

    This study reports the occurrence of different Vibrio and Salmonella species in 52 samples of Mytilus galloprovincialis collected from four sites along the Atlantic coast between Agadir and Essaouira (Anza, Cap Ghir, Imssouane and Essaouira). The level of Escherichia coli (E. coli) was also determined to evaluate the degree of microbial pollution in the investigated areas. In this study three methods were used : AFNOR NF EN ISO 6579 V08-013 for Salmonella spp., the provisional method routinely used by several laboratories (Institut Pasteur, Paris,…) for Vibrio cholerae and Vibrio parahaemolyticus in the seafood, and the most probable number method (MPN) using Norm ISO/TS 16649-3 (2005) for E. coli. The most frequently isolated Vibrios were Vibrio alginolyticus (90.4% of samples), followed by V. cholerae non O1 non O139 (15.4%) and V. parahaemolyticus (7.7%). Salmonella spp. was found in 15% of the samples. The number of E. coli ranged between 0.2/100 g and 1.8 10(3) /100 g of mussel soft tissues. This study indicates the potential sanitary risk associated with the presence of pathogenic bacteria in cultivated mussels in the two populous regions of southern Morocco, where shellfish production and maritime tourism are important to the local economy.

  3. Continuous Exposure Of Vibrio Anguillarum To Tropodithietic Acid: Genetic Changes And Influence On Virulence

    DEFF Research Database (Denmark)

    Rasmussen, Bastian Barker; D'Alvise, Paul; Grotkjær, Torben

    2015-01-01

    Introduction: The fish pathogen Vibrio anguillarum is a major problem in aquaculture causing Vibriosis. Bacteria of the Roseobacter clade can antagonize pathogenic vibrios in cultures in live feed such as microalgae, rotifers and Artemia, as well as in fish larvae. Therefore, roseobacters could...

  4. Sensitivity of Vibrio cholerae cells to lethal and mutagenic effect of UV-irradiation mediated by plasmids

    International Nuclear Information System (INIS)

    Tiganova, I.G.; Evdokimova, N.M.; Aleshkin, G.I.

    1988-01-01

    The effect of UV-irradiation on Vibrio cholerae cells and its changes mediated by the plasmid R245 have been studied. Vibrio cholerae strains 569B and RV31 have been shown to be considerably more sensitive to lethal effect of UV-irradiation as compared with Escherichia coli and Salmonella typhimurium cells. Highly toxigenic strain 569B and practically atoxigenic strain RV31 have the same UV-sensitivity. Lethla effect of UV-irradiation on Vibrio cholerae cells is incresed when the irradiated cells are plated on enriched media. UV-induction of mutations was not registered in plasmidless strains of Vibrio cholerae. Plasmid R245 increase UV-resistance of vibrio cells and makes them UV-mutable

  5. α-Hydroxyketone Synthesis and Sensing by Legionella and Vibrio

    Directory of Open Access Journals (Sweden)

    Hubert Hilbi

    2012-03-01

    Full Text Available Bacteria synthesize and sense low molecular weight signaling molecules, termed autoinducers, to measure their population density and community complexity. One class of autoinducers, the α-hydroxyketones (AHKs, is produced and detected by the water-borne opportunistic pathogens Legionella pneumophila and Vibrio cholerae, which cause Legionnaires’ disease and cholera, respectively. The “Legionella quorum sensing” (lqs or “cholera quorum sensing” (cqs genes encode enzymes that produce and sense the AHK molecules “Legionella autoinducer-1” (LAI-1; 3-hydroxypentadecane-4-one or cholera autoinducer-1 (CAI-1; 3-hydroxytridecane-4-one. AHK signaling regulates the virulence of L. pneumophila and V. cholerae, pathogen-host cell interactions, formation of biofilms or extracellular filaments, expression of a genomic “fitness island” and competence. Here, we outline the processes, wherein AHK signaling plays a role, and review recent insights into the function of proteins encoded by the lqs and cqs gene clusters. To this end, we will focus on the autoinducer synthases catalysing the biosynthesis of AHKs, on the cognate trans-membrane sensor kinases detecting the signals, and on components of the down-stream phosphorelay cascade that promote the transmission and integration of signaling events regulating gene expression.

  6. 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.

  7. Structure of Vibrio cholerae ribosome hibernation promoting factor

    International Nuclear Information System (INIS)

    De Bari, Heather; Berry, Edward A.

    2013-01-01

    The X-ray crystal structure of ribosome hibernation promoting factor from V. cholerae has been determined at 2.0 Å resolution. The crystal was phased by two-wavelength MAD using cocrystallized cobalt. The X-ray crystal structure of ribosome hibernation promoting factor (HPF) from Vibrio cholerae is presented at 2.0 Å resolution. The crystal was phased by two-wavelength MAD using cocrystallized cobalt. The asymmetric unit contained two molecules of HPF linked by four Co atoms. The metal-binding sites observed in the crystal are probably not related to biological function. The structure of HPF has a typical β–α–β–β–β–α fold consistent with previous structures of YfiA and HPF from Escherichia coli. Comparison of the new structure with that of HPF from E. coli bound to the Thermus thermophilus ribosome [Polikanov et al. (2012 ▶), Science, 336, 915–918] shows that no significant structural changes are induced in HPF by binding

  8. The pathogenesis, detection and prevention of Vibrio parahaemolyticus

    Directory of Open Access Journals (Sweden)

    Rongzhi eWang

    2015-03-01

    Full Text Available Vibrio parahaemolyticus, a Gram-negative motile bacterium that inhabits marine and estuarine environments throughout the world, is a major food-borne pathogen that causes life-threatening diseases in humans after the consumption of raw or undercooked seafood. The global occurrence of V. parahaemaolyticus accentuates the importance of investigating its virulence factors and their effects on the human host. This review describes the virulence factors of V. parahaemolyticus reported to date, including hemolysin, urease, two type III secretion systems (T3SS and two type VI secretion systems (T6SS, which both cause both cytotoxicity in cultured cells and enterotoxicity in animal models. We describe various types of detection methods, based on virulence factors, that are used for quantitative detection of V. parahaemolyticus in seafood. We also discuss some useful preventive measures and therapeutic strategies for the diseases mediated by V. parahaemolyticus, which can reduce, to some extent, the damage to humans and aquatic animals attributable to V. parahaemolyticus. This review extends our understanding of the pathogenic mechanisms of V. parahaemolyticus mediated by virulence factors and the diseases it causes in its human host. It should provide new insights for the diagnosis, treatment, and prevention of V. parahaemolyticus infection.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Genetic diversity and population structure of Vibrio cholerae.

    Science.gov (United States)

    Beltrán, P; Delgado, G; Navarro, A; Trujillo, F; Selander, R K; Cravioto, A

    1999-03-01

    Multilocus enzyme electrophoresis (MLEE) of 397 Vibrio cholerae isolates, including 143 serogroup reference strains and 244 strains from Mexico and Guatemala, identified 279 electrophoretic types (ETs) distributed in two major divisions (I and II). Linkage disequilibrium was demonstrated in both divisions and in subdivision Ic of division I but not in subdivision Ia, which includes 76% of the ETs. Despite this evidence of relatively frequent recombination, clonal lineages may persist for periods of time measured in at least decades. In addition to the pandemic clones of serogroups O1 and O139, which form a tight cluster of four ETs in subdivision Ia, MLEE analysis identified numerous apparent clonal lineages of non-O1 strains with intercontinental distributions. A clone of serogroup O37 that demonstrated epidemic potential in the 1960s is closely related to the pandemic O1/O139 clones, but the nontoxigenic O1 Inaba El Tor reference strain is not. A strain of serogroup O22, which has been identified as the most likely donor of exogenous rfb region DNA to the O1 progenitor of the O139 clone, is distantly related to the O1/O139 clones. The close evolutionary relationships of the O1, O139, and O37 epidemic clones indicates that new cholera clones are likely to arise by the modification of a lineage that is already epidemic or is closely related to such a clone.

  11. Non-01 Vibrio cholerae infections in Cancun, Mexico.

    Science.gov (United States)

    Finch, M J; Valdespino, J L; Wells, J G; Perez-Perez, G; Arjona, F; Sepulveda, A; Bessudo, D; Blake, P A

    1987-03-01

    To determine the role of Vibrio cholerae as a cause of diarrheal illness in Cancun, Mexico, an investigation was conducted in July and August 1983. Although toxigenic V. cholerae 01 were not found, non-01 V. cholerae were isolated from 22 (16%) of 134 stools from persons with diarrheal illness and none of 22 stools from well persons; 58 (92%) of 63 sewage samples; 12 (86%) of 14 untreated well water samples; a home storage tank for treated water; and 5 (21%) of 24 samples of raw seafood. None of the V. cholerae isolates from patients were toxigenic. The illness occurred mainly in small children, and were characterized principally by diarrhea and abdominal pain. No patient was seriously ill, and all recovered without sequelae. Seven different serotypes of non-01 V. cholerae were isolated from the stool specimens, and Smith serotype 12 accounted for 10 (46%) of the 22 isolates. A matched-pair case-control study found that cases were more likely than controls to have eaten home prepared gelatin (P = 0.03, OR = 5/0) and seafood (P = 0.06, OR = 4/0).

  12. [The in vitro action of plants on Vibrio cholerae].

    Science.gov (United States)

    Guevara, J M; Chumpitaz, J; Valencia, E

    1994-01-01

    Natural products of several plants, according to the geographic location, are used by Peruvian people in the popular treatment of diarrhea, with good success. When cholerae cases appeared in Peru, we were interested to know the "in vitro" effect against Vibrio cholerae 01, of these useful plants to treat diarrhea. The following plants were tested: Cichorium intybus, Althaea officinalis, Psorela glandulosa, Geranium maculatum, Punica granatum, Malus sativa, Cydonia oblonga, Chenopodium ambrosoides, Krameria triandria, Tea chinensis, Daucus carota, Persea gratissima, Psidium guayaba and Lippia dulcis. Decoction or infusion of the plants were used in the "in vitro" experiments. The following plants showed no "in vitro" effect against V. cholerae: Cichorium intybus, Althaea officinalis, Psorela glandulosa, Geranium maculatum, Chenopodium ambrosoides, Krameria triandria, Psidium guayaba, Lippia dulcis and Daucus carota. Decoction of Malus sativa and Cydenia oblonga showed bactericidal effect for their acidity and stone avocado (Persea gratissima) a late bactericidal effect. Tea infusión and the decoction of Punica granatum peel, showed the best bactericidal effect and we suggest to use them as to stop cholera spreading.

  13. Vibrio infections among marine and fresh-water fish

    Science.gov (United States)

    1959-01-01

    In 1951. B. J. Earpio found a vibrio infection among salmon fingerlings being reared in saltwater at the Deception Pass Biological Station of the Washington State Department of Fisheries. The disease waa characterized by erythema at the base of fins and on the sides of the fish, necrotic areas in the Inusculature, inflammation of the intestinal tract, and general septicernia. The disease reappeared the next year, killing nearly all of the churn salmon (Oncorhynchus keta) fingerlings, killing about half of the pink salmon (O. gorbuscha) fingerlings, and affecting to a lesser degree the chinook salmon (O. tshawytscha) fingerlings. Also, late in 1952. R. R. Rucker and E. J. Ordal found the same disease at a rainbow trout hatchery of the Washington State Department of Game at Vancouver. The disease caused severe losses there among the rainbow trout (Salmo gairdneri) and among the sea-run form of the same species (called steelhead trout). The disease was manifested by bloody, necrotic areas in the musculature and inflammation of the viscera, відоіШат to furumaculoвiв,

  14. Human Gut Microbiota Predicts Susceptibility to Vibrio cholerae Infection.

    Science.gov (United States)

    Midani, Firas S; Weil, Ana A; Chowdhury, Fahima; Begum, Yasmin A; Khan, Ashraful I; Debela, Meti D; Durand, Heather K; Reese, Aspen T; Nimmagadda, Sai N; Silverman, Justin D; Ellis, Crystal N; Ryan, Edward T; Calderwood, Stephen B; Harris, Jason B; Qadri, Firdausi; David, Lawrence A; LaRocque, Regina C

    2018-04-12

    Cholera is a public health problem worldwide and the risk factors for infection are only partially understood. We prospectively studied household contacts of cholera patients to compare those who were infected with those who were not. We constructed predictive machine learning models of susceptibility using baseline gut microbiota data. We identified bacterial taxa associated with susceptibility to Vibrio cholerae infection and tested these taxa for interactions with V. cholerae in vitro. We found that machine learning models based on gut microbiota predicted V. cholerae infection as well as models based on known clinical and epidemiological risk factors. A 'predictive gut microbiota' of roughly 100 bacterial taxa discriminated between contacts who developed infection and those who did not. Susceptibility to cholera was associated with depleted levels of microbes from the phylum Bacteroidetes. By contrast, a microbe associated with cholera by our modeling framework, Paracoccus aminovorans, promoted the in vitro growth of V. cholerae. Gut microbiota structure, clinical outcome, and age were also linked. These findings support the hypothesis that abnormal gut microbial communities are a host factor related to V. cholerae susceptibility.

  15. Systematic analysis of the lysine acetylome in Vibrio parahemolyticus.

    Science.gov (United States)

    Pan, Jianyi; Ye, Zhicang; Cheng, Zhongyi; Peng, Xiaojun; Wen, Liangyou; Zhao, Fukun

    2014-07-03

    Lysine acetylation of proteins is a major post-translational modification that plays an important regulatory role in almost every aspect of cells, both eukaryotes and prokaryotes. Vibrio parahemolyticus, a model marine bacterium, is a worldwide cause of bacterial seafood-borne illness. Here, we conducted the first lysine acetylome in this bacterium through a combination of highly sensitive immune-affinity purification and high-resolution LC-MS/MS. Overall, we identified 1413 lysine acetylation sites in 656 proteins, which account for 13.6% of the total proteins in the cells; this is the highest ratio of acetyl proteins that has so far been identified in bacteria. The bioinformatics analysis of the acetylome showed that the acetylated proteins are involved in a wide range of cellular functions and exhibit diverse subcellular localizations. More specifically, proteins related to protein biosynthesis and carbon metabolism are the preferential targets of lysine acetylation. Moreover, two types of acetylation motifs, a lysine or arginine at the +4/+5 positions and a tyrosine, histidine, or phenylalanine at the +1/+2 positions, were revealed from the analysis of the acetylome. Additionally, protein interaction network analysis demonstrates that a wide range of interactions are modulated by protein acetylation. This study provides a significant beginning for the in-depth exploration of the physiological role of lysine acetylation in V. parahemolyticus.

  16. Metabolic responses of Haliotis diversicolor to Vibrio parahaemolyticus infection.

    Science.gov (United States)

    Lu, Jie; Shi, Yanyan; Cai, Shuhui; Feng, Jianghua

    2017-01-01

    Vibrio parahemolyticus is a devastating bacterial pathogen that often causes outbreak of vibriosis in abalone Haliotis diversicolor. Elucidation of metabolic mechanisms of abalones in responding to V. parahemolyticus infection is essential for controlling the epidemic. In this work, 1 H NMR-based metabolomic techniques along with correlation and network analyses are used to investigate characteristic metabolites, as well as corresponding disturbed pathways in hepatopancreas and gill of H. diversicolor after V. parahemolyticus infection for 48 h. Results indicate that obvious gender- and tissue-specific metabolic responses are induced. Metabolic responses in female abalones are more clearly observed than those in males, which are primarily manifested in the accumulation of branched-chain amino acids and the depletion of organic osmolytes (homarine, betaine and taurine) in the infected gills of female abalones, as well as in the depletion of glutamate, branched-chain and aromatic amino acids in the infected hepatopancreases of female abalones. Moreover, based on major metabolic functions of the characteristic metabolites, we have found that V. parahemolyticus infection not only cause the disturbance in energy metabolism, nucleotide metabolism and osmotic balance, but also induce oxidative stress, immune stress and neurotoxic effect in different tissues with various mechanisms. Our study provides details of metabolic responses of abalones to V. parahemolyticus infection and will shed light on biochemical defence mechanisms of male and female hosts against pathogen infection. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. A conserved tad pilus promotes Vibrio vulnificus oyster colonization.

    Science.gov (United States)

    Pu, Meng; Duriez, Patrick; Arazi, Mattan; Rowe-Magnus, Dean A

    2018-02-01

    Vibrio vulnificus has the highest death rate (>35%) and per-case economic burden ($3.3 million) of any foodborne pathogen in the United States. Infections occur via open wounds or following ingestion of contaminated seafood, most infamously oysters. We isolated a 1000th generation descendant, designated NT that exhibited increased biofilm and aggregate formation relative to its parent. We identified two significant causal changes underlying these phenotypes. First, the entire 24-kb capsular polysaccharide biosynthesis locus, which is essential for virulence but inhibits biofilm formation, had been purged from the genome. However, NT formed more extensive biofilms and aggregates than a defined cps mutant, suggesting that additional factor(s) contributed to its phenotypes. Second, the expression of a tight adherence (tad) pilus locus was elevated in NT. Deletion of the associated pilin (flp) decreased NT biofilm and aggregate formation. Furthermore, NTΔflp strains were deficient relative to NT in an oyster colonization model, demonstrating a positive correlation between the biofilm and aggregation phenotypes associated with Tad pilus production and efficient bacterial retention by feeding oysters. Despite being widely distributed in the Vibrionaceae, this is the first demonstration of a bona fide physiological role for a Tad pilus in this bacterial family. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  18. Population dynamics of Vibrio fischeri during infection of Euprymna scolopes.

    Science.gov (United States)

    McCann, Jessica; Stabb, Eric V; Millikan, Deborah S; Ruby, Edward G

    2003-10-01

    The luminous bacterium Vibrio fischeri colonizes a specialized light-emitting organ within its squid host, Euprymna scolopes. Newly hatched juvenile squid must acquire their symbiont from ambient seawater, where the bacteria are present at low concentrations. To understand the population dynamics of V. fischeri during colonization more fully, we used mini-Tn7 transposons to mark bacteria with antibiotic resistance so that the growth of their progeny could be monitored. When grown in culture, there was no detectable metabolic burden on V. fischeri cells carrying the transposon, which inserts in single copy in a specific intergenic region of the V. fischeri genome. Strains marked with mini-Tn7 also appeared to be equivalent to the wild type in their ability to infect and multiply within the host during coinoculation experiments. Studies of the early stages of colonization suggested that only a few bacteria became associated with symbiotic tissue when animals were exposed for a discrete period (3 h) to an inoculum of V. fischeri cells equivalent to natural population levels; nevertheless, all these hosts became infected. When three differentially marked strains of V. fischeri were coincubated with juvenile squid, the number of strains recovered from an individual symbiotic organ was directly dependent on the size of the inoculum. Further, these results indicated that, when exposed to low numbers of V. fischeri, the host may become colonized by only one or a few bacterial cells, suggesting that symbiotic infection is highly efficient.

  19. Detection of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa)

    Science.gov (United States)

    Ekawati, ER; Yusmiati, S. N. H.

    2018-01-01

    Blood cockle (Anadara granosa) has high level of zinc and protein, which is beneficial for therapeutic function for malnourished particularly stunting case in children. Zinc in animal foods is more absorbable than that from vegetable food. Blood cockle (Anadara granosa) is rich in nutrient and an excellent environment for the growth of microorganisms. This research aimed to identify the contamination of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa). This was observation research with laboratory analysis. Salmonella sp. and Vibrio sp. were detected from blood cockle. Total plate count was determine of the total amount of the bacteria. Results detected from 20 samples of blood cockle showed that all samples were negative of Salmonella sp. and 1 sample positive Vibrio sp. The result of total plate count bacteria was < 5 x 105 colony/g sample.

  20. Occurrence of Vibrio parahaemolyticus and Vibrio vulnificus in retail raw oysters from the eastern coast of Thailand.

    Science.gov (United States)

    Changchai, Nuttawee; Saunjit, Sudarat

    2014-05-01

    Occurrence, population density and virulence of Vibrio parahaemolyticus and V. vulnificus in 240 retail raw oysters collected monthly between March 2010 and February 2011 from Ang Sila coast, Chon Buri Province, Thailand were determined using most probable number (MPN) multiplex PCR. Multiplex PCR detected V. parahaemolyticus in 219 raw oyster samples, of which 29 samples contained the virulence tdh. MPN values for V. parahaemolyticus and pathogenic strains in most samples ranged from 10 to 10(2) and from 3 to 10 MPN/g, respectively. The presence of V. vulnificus was found in 53 oyster samples in amounts between 10 and 10(2) MPN/g. Of 1,087 V. parahaemolyticus isolates, 14 and 2 isolates carried tdh and virulence trh, respectively but none with both genes. However, none of the presumptive isolates was shown to be V. vulnificus. The detection of pathogenic V. parahaemolyticus and V. vulnificus in raw oysters has rendered high awareness of risk in consumption of raw or undercooked oysters.

  1. 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.

  2. Structural requirements of cholesterol for binding to Vibrio cholerae hemolysin.

    Science.gov (United States)

    Ikigai, Hajime; Otsuru, Hiroshi; Yamamoto, Koichiro; Shimamura, Tadakatsu

    2006-01-01

    Cholesterol is necessary for the conversion of Vibrio cholerae hemolysin (VCH) monomers into oligomers in liposome membranes. Using different sterols, we determined the stereochemical structures of the VCH-binding active groups present in cholesterol. The VCH monomers are bound to cholesterol, diosgenin, campesterol, and ergosterol, which have a hydroxyl group at position C-3 (3betaOH) in the A ring and a C-C double bond between positions C-5 and C-6 (C-C Delta(5)) in the B ring. They are not bound to epicholesterol and dihydrocholesterol, which form a covalent link with a 3alphaOH group and a C-C single bond between positions C-5 and C-6, respectively. This result suggests that the 3betaOH group and the C-CDelta(5) bond in cholesterol are required for VCH monomer binding. We further examined VCH oligomer binding to cholesterol. However, this oligomer did not bind to cholesterol, suggesting that the disappearance of the cholesterol-binding potential of the VCH oligomer might be a result of the conformational change caused by the conversion of the monomer into the oligomer. VCH oligomer formation was observed in liposomes containing sterols with the 3betaOH group and the C-C Delta(5) bond, and it correlated with the binding affinity of the monomer to each sterol. Therefore, it seems likely that monomer binding to membrane sterol leads to the assembly of the monomer. However, since oligomer formation was induced by liposomes containing either epicholesterol or dihydrocholesterol, the 3betaOH group and the C-C Delta(5) bond were not essential for conversion into the oligomer.

  3. 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

  4. Vibrio cholerae Infection of Drosophilamelanogaster Mimics the Human Disease Cholera.

    Directory of Open Access Journals (Sweden)

    2005-09-01

    Full Text Available Cholera, the pandemic diarrheal disease caused by the gram-negative bacterium Vibrio cholerae, continues to be a major public health challenge in the developing world. Cholera toxin, which is responsible for the voluminous stools of cholera, causes constitutive activation of adenylyl cyclase, resulting in the export of ions into the intestinal lumen. Environmental studies have demonstrated a close association between V. cholerae and many species of arthropods including insects. Here we report the susceptibility of the fruit fly, Drosophila melanogaster, to oral V. cholerae infection through a process that exhibits many of the hallmarks of human disease: (i death of the fly is dependent on the presence of cholera toxin and is preceded by rapid weight loss; (ii flies harboring mutant alleles of either adenylyl cyclase, Gsalpha, or the Gardos K channel homolog SK are resistant to V. cholerae infection; and (iii ingestion of a K channel blocker along with V. cholerae protects wild-type flies against death. In mammals, ingestion of as little as 25 mug of cholera toxin results in massive diarrhea. In contrast, we found that ingestion of cholera toxin was not lethal to the fly. However, when cholera toxin was co-administered with a pathogenic strain of V. cholerae carrying a chromosomal deletion of the genes encoding cholera toxin, death of the fly ensued. These findings suggest that additional virulence factors are required for intoxication of the fly that may not be essential for intoxication of mammals. Furthermore, we demonstrate for the first time the mechanism of action of cholera toxin in a whole organism and the utility of D. melanogaster as an accurate, inexpensive model for elucidation of host susceptibility to cholera.

  5. Vibrio Parahemolyticus in the Wastewater of Kermanshah City

    OpenAIRE

    Ali Almasi; Mohammad Soltanian; Feizollah Mansoori; Mohammad Taghi Aivazy; Parviz Mohajeri; Habibeh Meskini

    2005-01-01

    آب و فاضلاب                                                                                                                                                                                                               شماره 51- سال 1383     Municipal wastewater is one of the most important pollution sources for water supply resources. Soil, vegetable, and food material are exposed as well. Identification and enumeration of pathogenic agents particularly pathogenic Vibrios a...

  6. Modeling Transfer of Vibrio Parahaemolyticus During Peeling of Raw Shrimp.

    Science.gov (United States)

    Xiao, Xingning; Pang, Haiying; Wang, Wen; Fang, Weihuan; Fu, Yingchun; Li, Yanbin

    2018-03-01

    This study aimed to qualify the transfer of Vibrio parahaemolyticus during the shrimp peeling process via gloves under 3 different scenarios. The 1st 2 scenarios provided quantitative information for the probability distribution of bacterial transfer rates from (i) contaminated shrimp (6 log CFU/g) to non-contaminated gloves (Scenario 1) and (ii) contaminated gloves (6 log CFU/per pair) to non-contaminated shrimp (Scenario 2). In Scenario 3, bacterial transfer from contaminated shrimp to non-contaminated shrimp in the shrimp peeling process via gloves was investigated to develop a predictive model for describing the successive bacterial transfer. The range of bacterial transfer rate (%) in Scenarios 1 and 2 was 7% to 91.95% and 0.04% to 12.87%, respectively, indicating that the bacteria can be transferred from shrimp to gloves much easier than that from gloves to shrimp. A Logistic (1.59, 0.14) and Triangle distribution (-1.61, 0.12, 1.32) could be used to describe the bacterial transfer rate in Scenarios 1 and 2, respectively. In Scenario 3, a continuously decay patterning with fluctuations as the peeling progressed has been observed at all inoculation levels of the 1st shrimp (5, 6, and 7 log CFU/g). The bacteria could be transferred easier at 1st few peels, and the decreasing bacterial transfer was found in later phase. Two models (exponential and Weibull) could describe the successive bacterial transfer satisfactorily (pseudo-R 2 > 0.84, RMSE peeling process. The bacterial transfer rate distribution and predictive model derived from this work could be used in risk assessment of V. parahaemolyticus to ensure peeled shrimp safety. © 2018 Institute of Food Technologists®.

  7. The annual cycle of zooplankton-associated Vibrio cholerae and related vibrios in Albufera lake and its coastal surrounding waters (Valencia, Spain).

    Science.gov (United States)

    Arnau, A; Pujalte, M J; Amaro, C; Garay, E

    1988-12-01

    Most probable numbers of zooplankton-associated Vibrio spp. and V. cholerae were determined in Albufera lake, Spain, and in its coastal receiving waters throughout a year. Highest counts of 10(5) bacterial cells/g of plankton were associated to high temperatures and were also related to the kind of water. All isolates were non-01 serovars, and most belonged to Heiberg groups I and II.

  8. Impact of solar irradiation on cholera toxin secretion by different strains of Vibrio cholerae

    CSIR Research Space (South Africa)

    Ssemakalu, CC

    2013-09-01

    Full Text Available , Salomon RN, Garrity K, Reveillaud I, Kopin A, Jackson FR, et al. Vibrio cholerae infection of Drosophila melanogaster mimics the human disease cholera. PLoS Pathog. 2005;1(1):e8. http://dx.doi.org/10.1371/ journal.ppat.0010008 2. World Health...

  9. [Post nearly Drowning Vibrio alginolyticus Septicemia Acquired in Reunion (Indian Ocean)].

    Science.gov (United States)

    Gaüzère, B-A; Chanareille, P; Vandroux, D

    2016-08-01

    AbstractWe report the first case of Vibrio alginolyticus septicemia in the Indian Ocean (Reunion Island), in a patient (70-year-old-man) with multiple underlying conditions, following a nearly drowning in the lagoon of Reunion. From now on, V. alginolyticus should be considered as a possible agent of septicemia in the Indian Ocean, particularly following marine activities.

  10. 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...

  11. Cerebral absces med Vibrio cholerae non-01 efter badning i dansk havvand

    DEFF Research Database (Denmark)

    Torp-Pedersen, Trine; Nielsen, Xiaohui Chen; Olsen, Katharina E P

    2012-01-01

    on the ankle. We treated the patient with meropenem and ciprofloxacin for six weeks followed by ciprofloxacin for four weeks, and she recovered with hemiparesis and speech impairment. Marine Vibrio species may produce intracranial infection in predisposed individuals, even in temperate climate zones....

  12. 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...

  13. Effects of triclosan on bacterial community composition and Vibrio populations in natural seawater microcosms

    OpenAIRE

    Lydon, Keri Ann; Glinski, Donna A.; Westrich, Jason R.; Henderson, W. Matthew; Lipp, Erin K.

    2017-01-01

    Pharmaceuticals and personal care products, including antimicrobials, can be found at trace levels in treated wastewater effluent. Impacts of chemical contaminants on coastal aquatic microbial community structure and pathogen abundance are unknown despite the potential for selection through antimicrobial resistance. In particular, 'Vibrio', a marine bacterial genus that includes several human pathogens, displays resistance to the ubiquitous antimicrobial compound triclosan. Here we demonstrat...

  14. Effects of triclosan on bacterial community composition and 'Vibrio' populations in natural seawater microcosms

    Directory of Open Access Journals (Sweden)

    Keri Ann Lydon

    2017-05-01

    Full Text Available Pharmaceuticals and personal care products, including antimicrobials, can be found at trace levels in treated wastewater effluent. Impacts of chemical contaminants on coastal aquatic microbial community structure and pathogen abundance are unknown despite the potential for selection through antimicrobial resistance. In particular, 'Vibrio', a marine bacterial genus that includes several human pathogens, displays resistance to the ubiquitous antimicrobial compound triclosan. Here we demonstrated through use of natural seawater microcosms that triclosan (at a concentration of ~5 ppm can induce a significant 'Vibrio' growth response (68–1,700 fold increases in comparison with no treatment controls for three distinct coastal ecosystems: Looe Key Reef (Florida Keys National Marine Sanctuary, Doctors Arm Canal (Big Pine Key, FL, and Clam Bank Landing (North Inlet Estuary, Georgetown, SC. Additionally, microbial community analysis by 16 S rRNA gene sequencing for Looe Key Reef showed distinct changes in microbial community structure with exposure to 5 ppm triclosan, with increases observed in the relative abundance of 'Vibrio'naceae (17-fold, Pseudoalteromonadaceae (65-fold, Alteromonadaceae (108-fold, Colwelliaceae (430-fold, and Oceanospirillaceae (1,494-fold. While the triclosan doses tested were above concentrations typically observed in coastal surface waters, results identify bacterial families that are potentially resistant to triclosan and/or adapted to use triclosan as a carbon source. The results further suggest the potential for selection of 'Vibrio' in coastal environments, especially sediments, where triclosan may accumulate at high levels.

  15. [Cytotoxic effect of Vibrio cholerae non-O1 on Vero cells].

    Science.gov (United States)

    Figueroa-Arredondo, P; García-Lozano, H; Gutiérrez-Cogco, L; Valdespino-Gómez, J L

    1994-01-01

    At the present time there is still in Mexico a diarrhoeal outbreak due to Vibrio cholerae O1. In INDRE we have isolated from the same outbreak last year (jan-apr), 70 strains of Vibrio cholerae Non-O1. These were isolated from patients with a diarrhoeal illness different from cholera. Patients were of different ages and sex, and from various geographic areas. The isolated strains were confirmed by serological agglutination test with polyclonal antisera, and they neither belong to O1 serogroup or O139. We assayed all the 70 strains in Vero cells, searching for cytotoxic effect, probably attributed to cholera toxin, or any other toxin. The strains were screened by PCR for cholera toxin gene detection, and negative results were obtained. We have found only one CT-producer strain, but it was a rough one so, we are not able to affirm that is not a V. cholerae O1 serotype. Vibrio cholerae Non-O1 strains, tested in Vero cells assay, produced cytotoxic effect within 24 h. It was found that 48/70 strains (66.6%), had cytotoxic activity, showing rounding and then lysis of cells. From our results we concluded that this cytotoxic effect, is not cholera toxin related, instead we propose it could be due to an unknown virulence factor, probably a different toxin in mexican Vibrio cholerae Non-O1 strains.

  16. The protective activity of tea catechins against experimental infection by Vibrio cholerae O1.

    Science.gov (United States)

    Toda, M; Okubo, S; Ikigai, H; Suzuki, T; Suzuki, Y; Hara, Y; Shimamura, T

    1992-01-01

    Tea catechins inhibited the fluid accumulation induced by cholera toxin in sealed adult mice. The catechins also reduced fluid accumulation by Vibrio cholerae O1 in ligated intestinal loops of rabbits. These findings suggest that tea catechins may possess protective activity against V. cholerae O1.

  17. Three draft genomes of Vibrio coralliilyticus strains isolated from bivalve hatcheries

    Science.gov (United States)

    Reported here are the draft genomes of three Vibrio coralliilyticus isolates RE87, AIC-7, and 080116A. Each strain was isolated in association with diseased oyster larvae in commercial aquaculture systems. These draft genomes will be useful for further studies in understanding the genomic features...

  18. Crystallization of the HigBA2 toxin-antitoxin complex from Vibrio cholerae

    DEFF Research Database (Denmark)

    Hadǽi, San; Garcia-Pino, Abel; Martinez-Rodriguez, Sergio

    2013-01-01

    The genome of Vibrio cholerae encodes two higBA toxin-antitoxin (TA) modules that are activated by amino-acid starvation. Here, the TA complex of the second module, higBA2, as well as the C-terminal domain of the corresponding HigA2 antitoxin, have been purified and crystallized. The HigBA2 compl...

  19. Isolation of Vibrio cholerae serotype O1 from the eastern oyster, Crassostrea virginica.

    Science.gov (United States)

    Hood, M A; Ness, G E; Rodrick, G E

    1981-01-01

    Two strains of Vibrio cholerae serotype O1 Inaba were isolated from eastern oysters, Crassostrea virginica, collected from estuarine waters in Florida during April 1980. The oyster meats and waters from which the oysters were collected had low fecal coliform counts, and the area had no prior evidence of sewage contamination. PMID:7235700

  20. 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...

  1. Structural organization of the transfer RNA operon I of Vibrio cholerae

    Indian Academy of Sciences (India)

    Nine major transfer RNA (tRNA) gene clusters were analysed in various Vibrio cholerae strains. Of these, only the tRNA operon I was found to differ significantly in V. cholerae classical (sixth pandemic) and El Tor (seventh pandemic) strains. Amongst the sixteen tRNA genes contained in this operon, genes for tRNA Gln3 ...

  2. INS refuerza vigilancia y diagnóstico de enfermedad diarreica producida por Vibrio parahemolyticus

    OpenAIRE

    Huapaya, Blanca

    2005-01-01

    El Instituto Nacional de Salud (INS) está fortaleciendo la vigilancia y el diagnóstico en el Perú del Vibrio parahaemolyticus, bacteria causante de una enfermedad diarreica aguda, que a inicios de este año produjo un brote de aproximadamente 10 mil casos en Chile.

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

    Science.gov (United States)

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

    2007-12-01

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

  4. 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 o...

  5. Determining Vaccination Frequency in Farmed Rainbow Trout Using Vibrio anguillarum O1 Specific Serum Antibody Measurements

    DEFF Research Database (Denmark)

    Holten-Andersen, Lars; Dalsgaard, Inger; Nylén, Jørgen

    2012-01-01

    Background Despite vaccination with a commercial vaccine with a documented protective effect against Vibrio anguillarum O1 disease outbreaks caused by this bacterium have been registered among rainbow trout at Danish fish farms. The present study examined specific serum antibody levels as a valid...

  6. 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 r...

  7. Occurrence and molecular characterisation of Vibrio parahaemolyticus in crustaceans commercialised in Venice area, Italy.

    Science.gov (United States)

    Caburlotto, Greta; Suffredini, Elisabetta; Toson, Marica; Fasolato, Luca; Antonetti, Paolo; Zambon, Michela; Manfrin, Amedeo

    2016-03-02

    Infections due to the pathogenic human vibrios, Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus, are mainly associated with consumption of raw or partially cooked bivalve molluscs. At present, little is known about the presence of Vibrio species in crustaceans and the risk of vibriosis associated with the consumption of these products. The aim of the present study was to evaluate the prevalence and concentration of the main pathogenic Vibrio spp. in samples of crustaceans (n=143) commonly eaten in Italy, taking into account the effects of different variables such as crustacean species, storage conditions and geographic origin. Subsequently, the potential pathogenicity of V. parahaemolyticus strains isolated from crustaceans (n=88) was investigated, considering the classic virulence factors (tdh and trh genes) and four genes coding for relevant proteins of the type III secretion systems 2 (T3SS2α and T3SS2β). In this study, the presence of V. cholerae and V. vulnificus was never detected, whereas 40 samples (28%) were positive for V. parahaemolyticus with an overall prevalence of 41% in refrigerated products and 8% in frozen products. The highest prevalence and average contamination levels were detected in Crangon crangon (prevalence 58% and median value 3400 MPN/g) and in products from the northern Adriatic Sea (35%), with the samples from the northern Venetian Lagoon reaching a median value of 1375 MPN/g. While genetic analysis confirmed absence of the tdh gene, three of the isolates contained the trh gene and, simultaneously, the T3SS2β genes. Moreover three possibly clonal tdh-negative/trh-negative isolates carried the T3SS2α apparatus. The detection of both T3SS2α and T3SS2β apparatuses in V. parahaemolyticus strains isolated from crustaceans emphasised the importance of considering new genetic markers associated with virulence besides the classical factors. Moreover this study represents the first report dealing with Vibrio spp. in

  8. Application of chitosan microparticles for reduction of vibrio species in seawater and live oysters (Crassostrea virginica).

    Science.gov (United States)

    Fang, Lei; Wolmarans, Bernhard; Kang, Minyoung; Jeong, Kwang C; Wright, Anita C

    2015-01-01

    Human Vibrio infections associated with consumption of raw shellfish greatly impact the seafood industry. Vibrio cholerae-related disease is occasionally attributed to seafood, but V. vulnificus and V. parahaemolyticus are the primary targets of postharvest processing (PHP) efforts in the United States, as they pose the greatest threat to the industry. Most successful PHP treatments for Vibrio reduction also kill the molluscs and are not suitable for the lucrative half-shell market, while nonlethal practices are generally less effective. Therefore, novel intervention strategies for Vibrio reduction are needed for live oyster products. Chitosan is a bioactive derivative of chitin that is generally recognized as safe as a food additive by the FDA, and chitosan microparticles (CMs) were investigated in the present study as a potential PHP treatment for live oyster applications. Treatment of broth cultures with 0.5% (wt/vol) CMs resulted in growth cessation of V. cholerae, V. vulnificus, and V. parahaemolyticus, reducing culturable levels to nondetectable amounts after 3 h in three independent experiments. Furthermore, a similar treatment in artificial seawater at 4, 25, and 37°C reduced V. vulnificus levels by ca. 7 log CFU/ml after 24 h of exposure, but 48 h of exposure and elevated temperature were required to achieve similar results for V. parahaemolyticus and V. cholerae. Live oysters that either were artificially inoculated or contained natural populations of V. vulnificus and V. parahaemolyticus showed significant and consistent reductions following CM treatment (5%) compared to the amounts in the untreated controls. Thus, the results strongly support the promising potential for the application of CMs as a PHP treatment to reduce Vibrio spp. in intact live oysters. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Evaluation of the natural prevalence of Vibrio spp. in Uruguayan mussels (Mytilus sp.) and their control using irradiation

    International Nuclear Information System (INIS)

    Lopez, C.

    2001-01-01

    The presence of potentially pathogenic bacteria belonging to the Vibrionacea, especially Vibrio cholerae, and of Salmonella spp., was examined in fresh Uruguayan mussels (Mytilus sp.) during two annual seasons. The radiation decimal reduction dose (D 10 ) of various toxigenic strains of Vibrio cholerae was determined to vary in vitro between 0.11 and 0.19 kGy. These results and those from the examination of natural Vibrio spp. contamination in mussels were used to conclude that 1.0 kGy would be enough to render Uruguayan mussels Vibrio-safe. Mussels irradiated in the shell at the optical dose survived long enough to allow the eventual introduction of irradiation as an effective intervention measure without affecting local marketing practices, and making it possible to market the fresh mussels live, as required by Uruguayan legislation. (author)

  10. 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.

  11. Exoproteome and Secretome Derived Broad Spectrum Novel Drug and Vaccine Candidates in Vibrio cholerae Targeted by Piper betel Derived Compounds

    Science.gov (United States)

    Barh, Debmalya; Barve, Neha; Gupta, Krishnakant; Chandra, Sudha; Jain, Neha; Tiwari, Sandeep; Leon-Sicairos, Nidia; Canizalez-Roman, Adrian; Rodrigues dos Santos, Anderson; Hassan, Syed Shah; Almeida, Síntia; Thiago Jucá Ramos, Rommel; Augusto Carvalho de Abreu, Vinicius; Ribeiro Carneiro, Adriana; de Castro Soares, Siomar; Luiz de Paula Castro, Thiago; Miyoshi, Anderson; Silva, Artur; Kumar, Anil; Narayan Misra, Amarendra; Blum, Kenneth; Braverman, Eric R.; Azevedo, Vasco

    2013-01-01

    Vibrio cholerae is the causal organism of the cholera epidemic, which is mostly prevalent in developing and underdeveloped countries. However, incidences of cholera in developed countries are also alarming. Because of the emergence of new drug-resistant strains, even though several generic drugs and vaccines have been developed over time, Vibrio infections remain a global health problem that appeals for the development of novel drugs and vaccines against the pathogen. Here, applying comparative proteomic and reverse vaccinology approaches to the exoproteome and secretome of the pathogen, we have identified three candidate targets (ompU, uppP and yajC) for most of the pathogenic Vibrio strains. Two targets (uppP and yajC) are novel to Vibrio, and two targets (uppP and ompU) can be used to develop both drugs and vaccines (dual targets) against broad spectrum Vibrio serotypes. Using our novel computational approach, we have identified three peptide vaccine candidates that have high potential to induce both B- and T-cell-mediated immune responses from our identified two dual targets. These two targets were modeled and subjected to virtual screening against natural compounds derived from Piper betel. Seven compounds were identified first time from Piper betel to be highly effective to render the function of these targets to identify them as emerging potential drugs against Vibrio. Our preliminary validation suggests that these identified peptide vaccines and betel compounds are highly effective against Vibrio cholerae. Currently we are exhaustively validating these targets, candidate peptide vaccines, and betel derived lead compounds against a number of Vibrio species. PMID:23382822

  12. Isolation of vibrio spp. In oysters (crassostrea rhizophorea) caught in the ‘de la virgen’ swamp

    OpenAIRE

    López Gutiérrez, Lercy; Autor; Manjarrez Pava, Ganiveth; Autor; Herrera Rodríguez, Lilibeth; Autor; Montes Payares, Ana Elena; Autor; Olascuaga Ruíz, Yuranis Paola; Autor; Ortega Quiroz, Rolando José; Autor

    2015-01-01

    Objective:  To establish contamination by Vibrio in oysters (Crassostrea rhizophorae) caught in De La Virgen Swamp, in order to alert entities in charge or protecting consumer health in Cartagena city. Methods: Between February and April 2006, 67 oysters from 5 strategic sites along De La Virgen Swamp, were analyzed. Insulation and identification of Vibrio was performed through a culture and biochemical tests.Results.  Predominant species were V. alginolyticus (23%),V fluvialis  (20%),V. para...

  13. Abundance and Multilocus Sequence Analysis of Vibrio Bacteria Associated with Diseased Elkhorn Coral (Acropora palmata) of the Florida Keys.

    Science.gov (United States)

    Kemp, Keri M; Westrich, Jason R; Alabady, Magdy S; Edwards, Martinique L; Lipp, Erin K

    2018-01-15

    The critically endangered elkhorn coral ( Acropora palmata ) is affected by white pox disease (WPX) throughout the Florida Reef Tract and wider Caribbean. The bacterium Serratia marcescens was previously identified as one etiologic agent of WPX but is no longer consistently detected in contemporary outbreaks. It is now believed that multiple etiologic agents cause WPX; however,