Quorum Sensing in Vibrio and Its Relevance to Bacterial Virulence
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Delineating Virulence of Vibrio Campbellii
www.nature.com/scientificreports OPEN Delineating virulence of Vibrio campbellii: a predominant luminescent bacterial pathogen in Indian shrimp hatcheries Sujeet Kumar1*, Chandra Bhushan Kumar1,2, Vidya Rajendran1, Nishawlini Abishaw1, P. S. Shyne Anand1, S. Kannapan1, Viswas K. Nagaleekar3, K. K. Vijayan1 & S. V. Alavandi1 Luminescent vibriosis is a major bacterial disease in shrimp hatcheries and causes up to 100% mortality in larval stages of penaeid shrimps. We investigated the virulence factors and genetic identity of 29 luminescent Vibrio isolates from Indian shrimp hatcheries and farms, which were earlier presumed as Vibrio harveyi. Haemolysin gene-based species-specifc multiplex PCR and phylogenetic analysis of rpoD and toxR identifed all the isolates as V. campbellii. The gene-specifc PCR revealed the presence of virulence markers involved in quorum sensing (luxM, luxS, cqsA), motility (faA, lafA), toxin (hly, chiA, serine protease, metalloprotease), and virulence regulators (toxR, luxR) in all the isolates. The deduced amino acid sequence analysis of virulence regulator ToxR suggested four variants, namely A123Q150 (AQ; 18.9%), P123Q150 (PQ; 54.1%), A123P150 (AP; 21.6%), and P123P150 (PP; 5.4% isolates) based on amino acid at 123rd (proline or alanine) and 150th (glutamine or proline) positions. A signifcantly higher level of the quorum-sensing signal, autoinducer-2 (AI-2, p = 2.2e−12), and signifcantly reduced protease activity (p = 1.6e−07) were recorded in AP variant, whereas an inverse trend was noticed in the Q150 variants AQ and PQ. The pathogenicity study in Penaeus (Litopenaeus) vannamei juveniles revealed that all the isolates of AQ were highly pathogenic with Cox proportional hazard ratio 15.1 to 32.4 compared to P150 variants; PP (5.4 to 6.3) or AP (7.3 to 14). -
Genetic Diversity of Culturable Vibrio in an Australian Blue Mussel Mytilus Galloprovincialis Hatchery
Vol. 116: 37–46, 2015 DISEASES OF AQUATIC ORGANISMS Published September 17 doi: 10.3354/dao02905 Dis Aquat Org Genetic diversity of culturable Vibrio in an Australian blue mussel Mytilus galloprovincialis hatchery Tzu Nin Kwan*, Christopher J. S. Bolch National Centre for Marine Conservation and Resource Sustainability, University of Tasmania, Locked Bag 1370, Newnham, Tasmania 7250, Australia ABSTRACT: Bacillary necrosis associated with Vibrio species is the common cause of larval and spat mortality during commercial production of Australian blue mussel Mytilus galloprovincialis. A total of 87 randomly selected Vibrio isolates from various stages of rearing in a commercial mus- sel hatchery were characterised using partial sequences of the ATP synthase alpha subunit gene (atpA). The sequenced isolates represented 40 unique atpA genotypes, overwhelmingly domi- nated (98%) by V. splendidus group genotypes, with 1 V. harveyi group genotype also detected. The V. splendidus group sequences formed 5 moderately supported clusters allied with V. splen- didus/V. lentus, V. atlanticus, V. tasmaniensis, V. cyclitrophicus and V. toranzoniae. All water sources showed considerable atpA gene diversity among Vibrio isolates, with 30 to 60% of unique isolates recovered from each source. Over half (53%) of Vibrio atpA genotypes were detected only once, and only 7 genotypes were recovered from multiple sources. Comparisons of phylogenetic diversity using UniFrac analysis showed that the culturable Vibrio community from intake, header, broodstock and larval tanks were phylogenetically similar, while spat tank communities were different. Culturable Vibrio associated with spat tank seawater differed in being dominated by V. toranzoniae-affiliated genotypes. The high diversity of V. splendidus group genotypes detected in this study reinforces the dynamic nature of microbial communities associated with hatchery culture and complicates our efforts to elucidate the role of V. -
16S Ribosomal DNA Sequencing Confirms the Synonymy of Vibrio Harveyi and V
DISEASES OF AQUATIC ORGANISMS Vol. 52: 39–46, 2002 Published November 7 Dis Aquat Org 16S ribosomal DNA sequencing confirms the synonymy of Vibrio harveyi and V. carchariae Eric J. Gauger, Marta Gómez-Chiarri* Department of Fisheries, Animal and Veterinary Science, 20A Woodward Hall, University of Rhode Island, Kingston, Rhode Island 02881, USA ABSTRACT: Seventeen bacterial strains previously identified as Vibrio harveyi (Baumann et al. 1981) or V. carchariae (Grimes et al. 1984) and the type strains of V. harveyi, V. carchariae and V. campbellii were analyzed by 16S ribosomal DNA (rDNA) sequencing. Four clusters were identified in a phylo- genetic analysis performed by comparing a 746 base pair fragment of the 16S rDNA and previously published sequences of other closely related Vibrio species. The type strains of V. harveyi and V. car- chariae and about half of the strains identified as V. harveyi or V. carchariae formed a single, well- supported cluster designed as ‘bona fide’ V. harveyi/carchariae. A second more heterogeneous clus- ter included most other strains and the V. campbellii type strain. Two remaining strains are shown to be more closely related to V. rumoiensis and V. mediterranei. 16S rDNA sequencing has confirmed the homogeneity and synonymy of V. harveyi and V. carchariae. Analysis of API20E biochemical pro- files revealed that they are insufficient by themselves to differentiate V. harveyi and V. campbellii strains. 16S rDNA sequencing, however, can be used in conjunction with biochemical techniques to provide a reliable method of distinguishing V. harveyi from other closely related species. KEY WORDS: Vibrio harveyi · Vibrio carchariae · Vibrio campbellii · Vibrio trachuri · Ribosomal DNA · Biochemical characteristics · Diagnostic · API20E Resale or republication not permitted without written consent of the publisher INTRODUCTION environmental sources (Ruby & Morin 1979, Orndorff & Colwell 1980, Feldman & Buck 1984, Grimes et al. -
Molecular Characterization of Vibrio Harveyi in Diseased Shrimp
Alexandria Journal of Veterinary Sciences www.alexjvs.com AJVS. Vol. 51(2): 358-366 November, 2016 DOI: 10.5455/ajvs.206029 Molecular Characterization of Vibrio Harveyi in Diseased Shrimp Helmy A. Torky1, Gaber S. Abdellrazeq1, Mona M. Hussein2, Nourhan H. Ghanem2 1Department of Microbiology, Faculty of Veterinary Medicine, Alexandria University, Egypt. 2Department of Fish Diseases, Animal health research institute, Egypt. Abstract Key words: The objective of this study was to characterize Vibrio harveyi phenotypically and by molecular methods. A total number of 420 shrimp post Molecular Characterization, larvae samples were collected, 280 samples of diseased cultured shrimp and 140 Vibrio harveyi, Diseased Shrimp. samples of apparently healthy marine shrimp. The samples were subjected to bacteriological examination. Thirty three (11.7%) of cultured shrimp samples and 4 (2.8%) of marine shrimp samples were phenotypically positive for V. harveyi. Species - specific gene (toxRgene) and two virulence associated genes (vhh - Correspondence to: Vibrio harveyi haemolysin, and partial hly- haemolysin gene) were investigated Nourhan Hamada Ghanem using conventional PCR. ([email protected]) This study proved that using of partial haemolysin gene in molecular characterization of V. harveyi is the fastest and most accurate method to identify all isolates of V. harveyi (highly pathogenic, moderately pathogenic and non- pathogenic strains) due to the presence of a single copy of haemolysin gene encoded in all isolates, although it is not in the same locus in the genome. 1. INTRODUCTION and Suwanto 2000) .Johnson and Shunk (1936) described V. harveyi as Achromobacter. Hendrie et Bacteria of the genus Vibrio have been specifically al., (1970) studied the systematic study of implicated as shrimp pathogens because they are bioluminescence; the species was later assigned to regularly found in high numbers during periods of the genera Lucibacterium. -
The Vibrio Core Group Induces Yellow Band Disease in Caribbean and Indo-Pacific Reef-Building Corals
Journal of Applied Microbiology ISSN 1364-5072 ORIGINALARTICLE The Vibrio core group induces yellow band disease in Caribbean and Indo-Pacific reef-building corals J.M. Cervino1, F.L. Thompson2, B. Gomez-Gil3, E.A. Lorence4, T.J. Goreau5, R.L. Hayes6, K.B. Winiarski-Cervino7, G.W. Smith8, K. Hughen9 and E. Bartels10 1 Pace University, Department of Biological Sciences, New York & Department of Geochemistry, Woods Hole Oceanographic Institute, Woods Hole, USA 2 Department of Genetics, Federal University of Rio de Janeiro, Brazil 3 CIAD, A.C. Mazatlan Unit for Aquaculture, Mazatlan, Mexico 4 Pace University, Department of Biological Sciences, New York, NY, USA 5 Global Coral Reef Alliance, Cambridge, MA, USA 6 Howard University, Washington DC, USA 7 Pew Institute for Ocean Science, New York, NY, USA 8 University of South Carolina Aiken, SC, USA 9 Department of Chemistry & Geochemistry, Woods Hole Oceanographic Institute, Woods Hole, MA, USA 10 Mote Marine Laboratory, Summerland Key, FL, USA Keywords Abstract cell division, pathogens, shell-fish, Vibrio, zooxanthellae. Aims: To determine the relationship between yellow band disease (YBD)- associated pathogenic bacteria found in both Caribbean and Indo-Pacific reefs, Correspondence and the virulence of these pathogens. YBD is one of the most significant coral J.M. Cervino, Pace University, Department of diseases of the tropics. Biological Sciences, 1 Pace Plaza, New York, Materials and Results: The consortium of four Vibrio species was isolated from NY 10038, USA. E-mail [email protected] YBD tissue on Indo-Pacific corals: Vibrio rotiferianus, Vibrio harveyi, Vibrio Present address alginolyticus and Vibrio proteolyticus. This consortium affects Symbiodinium J.M. -
Quorum Sensing in Vibrios and Cross-Species Activation Of
University of Wisconsin Milwaukee UWM Digital Commons Theses and Dissertations May 2013 Quorum Sensing in Vibrios and Cross-Species Activation of Bioluminescence Lux Genes by Vibrio Harveyi LuxR in an Arabinose-Inducible Escherichia Coli Expression System Anne Marie Wannamaker University of Wisconsin-Milwaukee Follow this and additional works at: https://dc.uwm.edu/etd Part of the Evolution Commons, Microbiology Commons, and the Molecular Biology Commons Recommended Citation Wannamaker, Anne Marie, "Quorum Sensing in Vibrios and Cross-Species Activation of Bioluminescence Lux Genes by Vibrio Harveyi LuxR in an Arabinose-Inducible Escherichia Coli Expression System" (2013). Theses and Dissertations. 177. https://dc.uwm.edu/etd/177 This Thesis is brought to you for free and open access by UWM Digital Commons. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of UWM Digital Commons. For more information, please contact [email protected]. QUORUM SENSING IN VIBRIOS AND CROSS-SPECIES ACTIVATION OF BIOLUMINESCENCE LUX GENES BY VIBRIO HARVEYI LUXR IN AN ARABINOSE-INDUCIBLE ESCHERICHIA COLI EXPRESSION SYSTEM by Anne Marie Wannamaker A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science in Biological Sciences at The University of Wisconsin-Milwaukee May 2013 ABSTRACT QUORUM SENSING IN VIBRIOS AND CROSS-SPECIES ACTIVATION OF BIOLUMINESCENCE LUX GENES BY VIBRIO HARVEYI LUXR IN AN ARABINOSE-INDUCIBLE ESCHERICHIA EXPRESSION SYSTEM by Anne Marie Wannamaker The University of Wisconsin-Milwaukee, 2013 Under the Supervision of Dr. Charles Wimpee Bacterial bioluminescence is observed in over twenty known species, primarily in the family Vibrionaceae. However, only Vibrio fischeri and Vibrio harveyi bioluminescence expression mechanisms are well studied. -
Molecular Identification of Vibrio Harveyi-Related Bacteria and Vibrio Owensii Sp
ResearchOnline@JCU This file is part of the following reference: Cano Gomez, Ana (2012) Molecular identification of Vibrio harveyi-related bacteria and Vibrio owensii sp. nov., pathogenic to larvae of the ornate spiny lobster Panulirus ornatus. PhD thesis, James Cook University. Access to this file is available from: http://eprints.jcu.edu.au/23845/ The author has certified to JCU that they have made a reasonable effort to gain permission and acknowledge the owner of any third party copyright material included in this document. If you believe that this is not the case, please contact [email protected] and quote http://eprints.jcu.edu.au/23845/ Molecular identification of Vibrio harveyi-related bacteria and Vibrio owensii sp. nov., pathogenic to larvae of the ornate spiny lobster Panulirus ornatus Thesis submitted by Ana CANO-GÓMEZ B.Sc. (University of Cádiz, Spain) M.Appl.Sc. Biotechnology (James Cook University, Australia) In February 2012 for the degree of Doctor of Philosophy in the School of Veterinary and Biomedical Sciences James Cook University STATEMENT OF ACCESS I, the undersigned, author of this work, understand that James Cook University will make this thesis available for use within the University Library and, via the Australian Digital Theses network, for use elsewhere. I understand that, as an unpublished work, a thesis has significant protection under the Copyright Act and; I do not wish to place any further restriction on access to this work. Ana Cano-Gómez Febuary 2012 STATEMENT OF SOURCES DECLARATION I declare that this thesis is my own work and has not been submitted in any form for another degree or diploma at any university or other institution of tertiary education. -
Status and Progress in Coral Reef Disease Research
DISEASES OF AQUATIC ORGANISMS Vol. 69: 1–7, 2006 Published March 23 Dis Aquat Org INTRODUCTION Status and progress in coral reef disease research Ernesto Weil1,*, Garriet Smith2, Diego L. Gil-Agudelo3 1Department of Marine Sciences, University of Puerto Rico, PO Box 3208, Lajas, Puerto Rico 00667 2Department of Biology and Geology, University of South Carolina Aiken, Aiken, South Carolina 29801, USA 3Instituto de Investigaciones Marinas y Costeras INVEMAR, Cerro Punto de Betín, Zona Portuaria, Santa Marta, Magdalena, Colombia ABSTRACT: Recent findings on the ecology, etiology and pathology of coral pathogens, host resis- tance mechanisms, previously unknown disease/syndromes and the global nature of coral reef dis- eases have increased our concern about the health and future of coral reef communities. Much of what has been discovered in the past 4 years is presented in this special issue. Among the significant findings, the role that various Vibrio species play in coral disease and health, the composition of the ‘normal microbiota’ of corals, and the possible role of viruses in the disease process are important additions to our knowledge. New information concerning disease resistance and vectors, variation in pathogen composition for both fungal diseases of gorgonians and black band disease across oceans, environmental effects on disease susceptibility and resistance, and temporal and spatial disease variations among different coral species is presented in a number of papers. While the Caribbean may still be the ‘disease hot spot’ for coral reefs, it is now clear that diseases of coral reef organisms have become a global threat to coral reefs and a major cause of reef deterioration. -
Environmental Regulation of Bioluminescence in Vibrio
ENVIRONMENTAL REGULATION OF BIOLUMINESCENCE IN VIBRIO FISCHERI ES114 by NOREEN LORETTA LYELL (Under the Direction of Eric V. Stabb) ABSTRACT The pheromone-mediated circuitry that governs bioluminescence in Vibrio fischeri is well understood; however, less is known about the environmental conditions that influence pheromone and light production. The environment has been shown to have a profound effect on luminescence in V. fischeri, as cells in symbiotic association with the Hawaiian bobtail squid, Euprymna scolopes, are ~1000-fold brighter than non- symbiotic cells despite reaching similar cell densities. In this dissertation, I show that luminescence is governed by a complex regulatory web and that certain environmental conditions mediate the regulation of pheromone circuitry. My first goal was to identify and characterize previously unidentified regulators that control luminescence in V. fischeri strain ES114. I helped develop and employed a transposon mutagenesis system to discover novel negative regulators of luminescence. In this study, I characterized twenty-eight independent luminescence-up mutants with insertions in 14 loci. This work revealed that environmental conditions such as inorganic phosphate and Mg2+ concentrations are integrated into the regulation of the pheromone-dependent lux system. Furthermore, I showed that competition between the LuxI- and AinS-generated pheromones is an important and density-dependent factor in the level of light produced by V. fischeri ES114 cells, such that C8-HSL inhibits luminescence in dense populations. The second goal of this work was to clarify the role of cAMP receptor protein (CRP) in luminescence. Attempts to study the effects of glucose on V. fischeri luminescence have been contradictory and inconclusive, possibly due to strain-specific effects. -
Identification and Characterization of Bacteria with Antibacterial Activities Isolated from Seahorses (Hippocampus Guttulatus)
The Journal of Antibiotics (2010) 63, 271–274 & 2010 Japan Antibiotics Research Association All rights reserved 0021-8820/10 $32.00 www.nature.com/ja NOTE Identification and characterization of bacteria with antibacterial activities isolated from seahorses (Hippocampus guttulatus) Jose´ L Balca´zar1,2, Sara Loureiro1, Yolanda J Da Silva1, Jose´ Pintado1 and Miquel Planas1 The Journal of Antibiotics (2010) 63, 271–274; doi:10.1038/ja.2010.27; published online 26 March 2010 Keywords: antagonism; marine bacteria; seahorses; Vibrio species Seahorse populations have declined in the last years, largely due to To test the ability of the isolates to inhibit growth of pathogenic overfishing and habitat destruction.1 Recent research efforts have, there- Vibrio strains (Table 1), we grew all isolates on suitable agar media at fore, been focused to provide a better biological knowledge of these 20 1C for 2–3 days. After incubation, we spotted a loop of each isolate species. We have recently studied, as part of the Hippocampus project, onto the surface of marine agar previously inoculated with overnight wild populations of seahorse (Hippocampus guttulatus)insomeareasof cultures of the indicator strain. Clear zones after overnight incubation the Spanish coast and established breeding programs in captivity.2 at 20 1C indicated the presence of antibacterial substances. With the development of intensive production methods, it has Bacterial isolates showing antagonistic activity against pathogenic become apparent that diseases can be a significant limiting factor. Vibrio strains were identified using the 16S rRNA gene, amplified from Vibrio species are among the most important bacterial pathogens of extracted genomic DNA with primers 27F and 907R and Taq DNA marine fish. -
Control of Aliivibrio Fischeri Luminescence and Decrease In
Biocontrol Science, 2018, Vol. 23, No. 3, 85-96 Original Control of Aliivibrio fischeri Luminescence and Decrease in Bioluminescence by Fungicides HITOMI KUWAHARA1, JUNKO NINOMIYA1,2, AND HIROSHI MORITA3* 1Graduate School of Environment Engineering, University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan 2National Institute of Technology, Oita College, 1666 maki, Oita city, Oita 870-0152, Japan 3Faculty of Environment Engineering, University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan Received 29 August, 2017/Accepted 11 January, 2018 Studies have reported that cell density, ultraviolet( UV) irradiation, and redox reactions, can induce bioluminescence in bacteria. Conversely, the relationship between seawater components and luminescence is not well understood. The efficacy of marine luminous bacteria as biosensors, and their reactivity to fungicides( for example postharvest pesticides) are also unknown. Therefore, we studied the relationship between the luminescence of Aliivibrio fischeri and the composition of artificial seawater media and analyzed the toxicity of fungicides using A. fischeri grown only with the elements essential to induce luminescence. Luminescence was activated in the presence of KCl, NaHCO3, and MgSO4. In addition, we cultivated A. fischeri with + - 2- other compounds, including K , HCO3 , and SO4 ions. These results suggested that A. fischeri + - 2- requires K , HCO3 , and SO4 ions to activate cell density-independent luminescence. Additionally, A. fischeri cultured in 2.81% NaCl solutions containing KCl, NaHCO3, and MgSO4 exhibited a decrease in luminescence in the presence of sodium ortho-phenylphenol at >10 ppm. This result suggests that A. fischeri can be used as a biosensor to detect the presence of sodium ortho-phenylphenol. -
Marine Microlights: the Luminous Marine Bacteria Peter Herring
Marine microlights: the luminous marine bacteria Peter Herring Luminous marine Luminous bacteria have had a long and the yellow component of the luminescence. Up to bacteria can cause Gdistinguished career, albeit much of it cloaked 18 °C the light is yellow; at higher temperatures it fish to glow and in mystery. Glowing meat, fish and shrimp is blue. seas to shine. were well known to our less-illuminated ancestors Single isolated bacteria do not glow, but colonies do. and in 1667 Robert Boyle discovered that their light Individual cells produce an ‘autoinducer’, which How do they do it? was reversibly extinguished in a vacuum. A practical, accumulates in the medium and at a critical How can we use if unwitting, demonstration of the role of micro- concentration triggers the production of luciferase, this amazing organisms was described in 1825 in a macabre thereby switching on the luminescence. Different phenomenon in experiment on the eerie light emitted by two dissected species have different inducers, but similar systems. biomedical bodies in a London anatomy school. The luminous Autoinduction of luminescence is an example of quorum science? material was scraped off and was then used to induce sensing, in which population density (signalled other corpses to glow. chemically) induces metabolic processes, including Luminous bacteria were grown in culture in the virulence, in many non-luminous bacteria. 1870s and 27 species had been described by 1900. The present tally of culturable marine species is about 10, Genetics of luminescence three of them assigned to the genus Photobacterium, Light emission from marine bacteria is easy to detect.