Isolation and characterization of Vibrio alginolyticus from cultured amphioxus Branchiostoma belcheri tsingtauense
Abstract
Amphioxus is an intermediary species bridging invertebrates and vertebrates used to study the development and evolution of species and genes. In this study, the dominant bacterial strain was isolated from tail lesions of cultured amphioxus and designated WW1. Challenge by intramuscular microinjection revealed the 50% lethal dose of WW1 to be 9 × 102 bacteria/g amphioxus body weight. Based on 16S rRNA gene sequence, physiological and biochemical characteristics, and species-specific collagenase gene marker, WW1 was identified as Vibrio alginolyticus. To our knowledge, this is the first report on the isolation of V. alginolyticus as pathogen from amphioxus. This study extends the known host range of V. alginolyticus and may provide information helpful in preventing and managing its occurrence in cultured animals.
Acknowledgements
This work was supported by The National Sciences Foundation of China (No. 30972270), the National High-Technology Research and Development Program (863 Program) of China (2008AA092600), and the Scientific and Technological Innovation Project financially supported by Qingdao National Laboratory for Marine Science and Technology (No. 2015ASKJ02).
References
Ausubel F.M., Brent R., Kingston R.E., Moore D.D., Seidman J.G., Smith J.A. & Struhl K. 1995. Short Protocols in Molecular Biology. John Wiley & Sons, New York.Suche in Google Scholar
Altschul S.F., Madden T.L., Schaeffer A.A., Zhang J., Zhang Z., Miller W. & Lipman D.J. 1997. Gapped BLAST and PSIBLAST: a new generation of protein database search programs. Nucleic Acids Res. 25: 3389–3402.10.1093/nar/25.17.3389Suche in Google Scholar PubMed PubMed Central
Anguiano-Beltrán C., Lizárraga-Partida M.L. & Searcy-Bernal R. 2004. Effect of Vibrio alginolyticus on larval survival of the blue mussel Mytilus galloprovincialis. Dis. Aquat. Organ. 59: 119–123.10.3354/dao059119Suche in Google Scholar PubMed
Austin B. & Austin D.A. 1999. Bacterial Fish Pathogens: Disease of Farmed and Wild Fish, 3rd Ed. Springer-Verlag, Berlin.Suche in Google Scholar
Balcázar J.L., Gallo-Bueno A., Planas M. & Pintado J. 2010. Isolation of Vibrio alginolyticus and Vibrio splendidus from captive-bred seahorses with disease symptoms. Antonie Van Leeuwenhoek 97: 207–210.10.1007/s10482-009-9398-4Suche in Google Scholar PubMed
Balebona M.C., Andreu M.J., Bordas M.A., Zorrilla I., Moriñigo M.A. & Borrego J.J. 1998. Pathogenicity of Vibrio alginolyticus from cultured gilt-head sea bream (Sparus aurata L.). Appl. Environ. Microbiol. 64: 4269–4275.10.1128/AEM.64.11.4269-4275.1998Suche in Google Scholar PubMed PubMed Central
Ben Kahla-Nakbi A., Chaieb K. & Bakhrouf A. 2009. Investigation of several virulence properties among Vibrio alginolyticus strains isolated from diseased cultured fish in Tunisia. Dis. Aquat. Organ. 86: 21–28.10.3354/dao02091Suche in Google Scholar PubMed
Ben Kahla-Nakbi A., Chaieb K., Besbes A., Zmantar T. & Bakhrouf A. 2006. Virulence and enterobacterial repetitive intergenic consensus PCR of Vibrio alginolyticus strains isolated from Tunisian cultured gilthead sea bream and sea bass outbreaks. Vet. Microbiol. 117: 321–327.10.1016/j.vetmic.2006.06.012Suche in Google Scholar PubMed
Brenner D.J., Krieg N.R. & Staley J.T. 2004a. Bergey’s Manual of Systematic Bacteriology, 2nd Ed., Vol. 2, Part B, p. 513.Suche in Google Scholar
Brenner D.J., Krieg N.R. & Staley J.T. 2004b. Bergey’s Manual of Systematic Bacteriology, 2nd Ed., Vol. 2, Part B, p. 560.Suche in Google Scholar
Cai J., Han H., Song Z., Li C. & Zhou J. 2006. Isolation and characterization of pathogenic Vibrio alginolyticus from diseased postlarval abalone, Haliotis diversicolor supertexta (Lischke). Aquacult. Res. 37: 1222–1226.10.1111/j.1365-2109.2006.01552.xSuche in Google Scholar
Cannon J.P., Haire R.N. & Litman G.W. 2002. Identification of diversified genes that contain immunoglobulin-like variable regions in a protochordate. Nat. Immunol. 3: 1200–1207.10.1038/ni849Suche in Google Scholar PubMed
Chun J., Lee J.H., Jung Y., Kim M., Kim S., Kim B.K. & Lim Y.W. 2007. EzTaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences. Int. J. Syst. Evol. Microbiol. 57: 2259–2261.10.1099/ijs.0.64915-0Suche in Google Scholar PubMed
Colwell R.R. & Grimes D.J. 1984. Vibrio diseases of marine fish populations. Helgol. Mar. Res. 37: 265–287.10.1007/BF01989311Suche in Google Scholar
Di Pinto A., Ciccarese G., Tantillo G., Catalano D. & Forte V.T. 2005. A collagenase-targeted multiplex PCR assay for identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus. J. Food Prot. 68: 150–153.10.4315/0362-028X-68.1.150Suche in Google Scholar
Fang Y.Q. 1987. Ecological habits and protection of amphioxus. Chin. J. Zool. 22: 41–45 (in Chinese).Suche in Google Scholar
Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39: 783–791.10.1111/j.1558-5646.1985.tb00420.xSuche in Google Scholar
Gans C. 1996. Study of lancelets: the first 200 years. Israel J. Zool. 42: S3–S11.Suche in Google Scholar
Gomez-Leon J., Villamil L., Lemos M.L., Novoa B. & Figueras A. 2005. Isolation of Vibrio alginolyticus and Vibrio splendidus from aquacultured carpet shell clam (Ruditapes decussatus) larvae associated with mass mortalities. Appl. Environ. Microbiol. 71: 98–104.10.1128/AEM.71.1.98-104.2005Suche in Google Scholar
González-Escalona N., Blackstone G.M. & DePaola A. 2006. Characterization of a Vibrio alginolyticus strain, isolated from Alaskan oysters, carrying a hemolysin gene similar to the thermostable direct hemolysin-related hemolysin gene (trh) of Vibrio parahaemolyticus. Appl. Environ. Microbiol. 72: 7925–7929.10.1128/AEM.01548-06Suche in Google Scholar
Huang G.H., Liu H., Han Y., Fan L.F., Zhang Q.F., Liu J.Z., Yu X.S., Zhang L.C., Chen S.W., Dong M.L., Wang L. & Xu A.L. 2007. Profile of acute immune response in Chinese amphioxus uponStaphylococcus aureus and Vibrio parahaemolyticus infection. Dis. Aquat. Organ. 31: 1013–1023.10.1016/j.dci.2007.01.003Suche in Google Scholar
Holland L.Z. 2014. Genomics, evolution and development of amphioxus and tunicates: the goldilocks principle. J. Exp. Zool. 324B: 342–352.10.1002/jez.b.22569Suche in Google Scholar
Holland L.Z. & Yu J.K. 2004. Cephalochordate (amphioxus) embryos: procurement, culture, and basic methods. Methods Cell Biol. 74: 195–215.10.1016/S0091-679X(04)74009-1Suche in Google Scholar
Holland N.D., Holland L.Z. & Holland P.W. 2015. Scenarios for the making of vertebrates. Nature 520: 450–455.10.1038/nature14433Suche in Google Scholar PubMed
Jukes T.H. & Cantor C.R. 1969. Evolution of protein molecules, pp. 21–132. In: Munro H.N. (ed.) Mammalian Protein Metabolism. Academic Press, New York.10.1016/B978-1-4832-3211-9.50009-7Suche in Google Scholar
Lane D.J. 1991. 16S/23S rRNA sequencing, pp. 115–175. In: Stackebrandt E. & Goodfellow M. (eds) Nucleic Acid Techniques in Bacterial Systematics. John Wiley & Sons, New York.Suche in Google Scholar
Lee D.Y., Moon S.Y., Lee S.O., Yang H., Lee H.J. & Lee M.S. 2008. Septic shock due to Vibrio alginolyticus in a cirrhotic patient: the first case in Korea. Yonsei Medical Journal 49: 329–332.10.3349/ymj.2008.49.2.329Suche in Google Scholar PubMed PubMed Central
Lee K.K., Yu S.R., Chen F.R., Yang T.I. & Liu P.C. 1996a. Virulence of Vibrio alginolyticus isolated from diseased tiger prawn, Penaeus monodon. Curr. Microbiol. 32: 229–231.10.1007/s002849900041Suche in Google Scholar PubMed
Lee K.K., Yu S.R., Yang T.I., Liu P.C. & Chen F.R. 1996b. Isolation and characterization of Vibrio alginolyticus isolated from diseased kuruma prawn, Penaeus japonicus. Lett. Appl. Microbiol. 22: 111–114.10.1111/j.1472-765X.1996.tb01121.xSuche in Google Scholar
Lhafi S.K. & Kühne M. 2007. Occurrence of Vibrio spp. in blue mussels (Mytilus edulis) from the German Wadden Sea. Int. J. Food Microbiol. 116: 297–300.10.1016/j.ijfoodmicro.2007.01.007Suche in Google Scholar PubMed
Li J. 2002. Vibrio alginolyticus: Pathogenicity and Its Immunological Control via Vaccination in Silver Sea Bream, Sparussarba. Ph.D. Thesis, The Chinese University of Hong Kong, Hong Kong, People’s Republic of China, 242 pp.Suche in Google Scholar
Liu H., Wang Q.Y., Liu Q., Cao X.D., Shi C.B. & Zhang Y.X. 2011. Roles of Hfq in the stress adaptation and virulence in fish pathogen Vibrio alginolyticus and its potential application as a target for live attenuated vaccine. Appl. Microbiol. Biotechnol. 91: 353–364.10.1007/s00253-011-3286-3Suche in Google Scholar PubMed
Liu P.C., Chen Y.C. & Lee K.K. 2001. Pathogenicity of Vibrio alginolyticus isolated from diseased small abalone Haliotis diversicolor supertexta. Microbios 104: 71–77.Suche in Google Scholar
Martins M.L., Mouriño J.L., Fezer G.F., BuglioneNeto C.C., Garcia P., Silva B.C., Jatobá A. & Vieira F. 2010. Isolation and experimental infection with Vibrio alginolyticus in the sea horse, Hippocampus reidi Ginsburg, 1933 (Osteichthyes: Syngnathidae) in Brazil. Braz. J. Biol. 70: 205–209.10.1590/S1519-69842010000100028Suche in Google Scholar
Nei M. & Kumar S. 2000. Molecular Evolution and Phylogenetics. Oxford University Press, New York.Suche in Google Scholar
Pang Q., Zhang S., Liu X. & Wu D. 2006. Humoral immune responses of amphioxus Branchiostoma belcheri to challenge with Escherichia coli. Fish Shellfish Immunol. 21: 139–145.10.1016/j.fsi.2005.11.001Suche in Google Scholar PubMed
Pang Q., Zhang S. & Zhao B. 2009. Induction of phenoloxidases in the humoral fluids of amphioxus Branchiostoma belcheri by Vibrio alginolyticus and Escherichia coli. Fish Shellfish Immunol. 26: 669–671.10.1016/j.fsi.2009.02.019Suche in Google Scholar PubMed
Pang Q., Zhang S. & Zhao B. 2010. Immune parameters in the humoral fluids of amphioxus Branchiostoma belcheri challenged with Vibrio alginolyticus. Fish Shellfish Immunol. 28: 232–234.10.1016/j.fsi.2009.09.017Suche in Google Scholar PubMed
Reed L.J. & Muench H. 1938. A simple method of estimating fifty percent endpoints. Am. J. Hyg. 27: 493–497.10.1093/oxfordjournals.aje.a118408Suche in Google Scholar
Reilly G.D., Reilly C.A., Smith E.G. & Baker-Austin C. 2011. Vibrio alginolyticus-associated wound infection acquired in British waters, Guernsey, July 2011. Euro Surveill. 16: Pii: 19994.10.2807/ese.16.42.19994-enSuche in Google Scholar
Saitou N. & Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4: 406–425.Suche in Google Scholar
Sato A., Mayer W.E. & Klein J. 2003. A molecule bearing an immunoglobulin-like V region of the CTX subfamily in amphioxus. Immunogenetics 55: 423–427.10.1007/s00251-003-0589-2Suche in Google Scholar PubMed
Selvin J. & Lipton A.P. 2003. Vibrio alginolyticus associated with white spot disease of Penaeus monodon. Dis. Aquat. Organ. 57: 147–150.10.3354/dao057147Suche in Google Scholar PubMed
Snoussi M., Noumi E., Usai D., Sechi L.A., Zanetti S. & Bakhrouf A. 2008. Distribution of some virulence related-properties of Vibrio alginolyticus strains isolated from Mediterranean seawater (Bay of Khenis, Tunisia): investigation of eight Vibrio cholerae virulence genes. World J. Microbiol. Biotechnol. 24: 2133–2141.10.1007/s11274-008-9719-1Suche in Google Scholar
Tamura K., Stecher G., Peterson D., Filipski A. & Kumar S. 2013. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol. Biol. Evol. 30: 2725–2729.10.1093/molbev/mst197Suche in Google Scholar PubMed PubMed Central
Thompson F.L., Austin B. & Swings J. 2006. The Biology of Vibrios. ASM Press, Washington.10.1128/9781555815714Suche in Google Scholar
Thompson F.L., Lida T. & Swings J. 2004. Biodiversity of vibrios. Microbiol. Mol. Biol. Rev. 68: 403–431.10.1128/MMBR.68.3.403-431.2004Suche in Google Scholar PubMed PubMed Central
Wang G., Zhang S. & Wang Z. 2009. Responses of alternative complement expression to challenge with different combinations of Vibrio anguillarum, Escherichia coli and Staphylococcus aureus: evidence for specific immune priming in amphioxus Branchiostoma belcheri. Fish Shellfish Immunol. 26: 33–39.10.1016/j.fsi.2008.09.018Suche in Google Scholar PubMed
Wong H.C., Ting S.H. & Shieh W.R. 1992. Incidence of toxigenic vibrios in foods available in Taiwan. J. Appl. Bacteriol. 73: 197–202.10.1111/j.1365-2672.1992.tb02978.xSuche in Google Scholar PubMed
Wu X., Zhang S., Wang Y., Zhang B., Qu Y. & Jiang X. 1994. Laboratory observations on spawning, fecundity and larval development of amphioxus (Branchiostoma belcherit singtauense). Chin. J. Oceanol. Limnol. 12: 289–294.10.1007/BF02850488Suche in Google Scholar
Yasui K., Urata M., Yamaguchi N., Ueda H. & Henmi Y. 2007. Laboratory culture of the Oriental lancelet Branchiostoma belcheri. Zool. Sci. 24: 514–520.10.2108/zsj.24.514Suche in Google Scholar PubMed
Yu C., Dong M., Wu X., Li S., Huang S., Su J., Wei J., Shen Y., Mou C., Xie X., Lin J., Yuan S., Yu X., Yu Y., Du J., Zhang S., Peng X., Xiang M. & Xu A. 2005. Genes “waiting” for recruitment by the adaptive immune system: the insights from amphioxus. J. Immunol. 174: 3493–3500.10.4049/jimmunol.174.6.3493Suche in Google Scholar PubMed
Zhang Z., Schwartz S., Wagner L. & Miller W. 2000. A greedy algorithm for aligning DNA sequences. J. Comput. Biol. 7: 203–214.10.1089/10665270050081478Suche in Google Scholar PubMed
Zhou Z.J., Pang H.Y., Ding Y., Cai J., Huang Y.C., Jian J.C. & Wu Z.H. 2013. VscO, a putative T3SS chaperone escort of Vibrio alginolyticus, contributes to virulence in fish and is a target for vaccine development. Fish Shellfish Immunol. 35: 1523–1531.10.1016/j.fsi.2013.08.017Suche in Google Scholar PubMed
Zorrilla I., Moriñigo M.A., Castro D., Balebona M.C. & Borrego J.J. 2003. Intraspecific characterization of Vibrio alginolyticus isolates recovered from cultured fish in Spain. J. Appl. Microbiol. 95: 1106–1116.10.1046/j.1365-2672.2003.02078.xSuche in Google Scholar PubMed
- Abbreviations
- NS
normal saline
- TCBS
thiosulphate citrate bile salts sucrose
©2016 Institute of Molecular Biology, Slovak Academy of Sciences
Artikel in diesem Heft
- Zoology
- Climate warming and invasive fish species: Will they replace native fish species in waters of temperate zones?
- Cellular and Molecular Biology
- Production of poly-3-hydroxybutyrate from mixed culture
- Cellular and Molecular Biology
- Molecular and immunological characterization of Lp1, the 34 kDa outer membrane lipoprotein of Pseudomonas aeruginosa
- Cellular and Molecular Biology
- Antibacterial activity of a microemulsion loaded with cephalosporin
- Cellular and Molecular Biology
- Isolation and characterization of Vibrio alginolyticus from cultured amphioxus Branchiostoma belcheri tsingtauense
- Cellular and Molecular Biology
- Somatic embryogenesis in Phlox paniculata – histological analysis
- Botany
- Overexpression of the novel Zygophyllum xanthoxylum C2H2-type zinc finger gene ZxZF improves drought tolerance in transgenic Arabidopsis and poplar
- Botany
- Vegetation diversity of the two Dinaric karstic rivers in Bosnia and Herzegovina
- Botany
- Diversity and spatial patterns of plant communities in volcanic temporary ponds of Sicily (Italy)
- Zoology
- Erythraeus (Erythraeus) pistacicus sp. n. (Trombidiformes: Erythraeidae) from southern Iran, and notes on other Erythraeus spp.
- Zoology
- Studies on eucrenal-hypocrenal zonation of springs along the river mainstream: A case study of a karst canyon in Bosnia and Herzegovina
- Zoology
- Different catching series from light and pheromone trapping of Helicoverpa armigera (Lepidoptera: Noctuidae)
- Zoology
- Newts in the nature reserve Dönche: Evaluation of diversity, breeding behaviour and threats due to a decline in habitat suitability
- Zoology
- Antioxidant status of rabbits after treatment with epicatechin and patulin
Artikel in diesem Heft
- Zoology
- Climate warming and invasive fish species: Will they replace native fish species in waters of temperate zones?
- Cellular and Molecular Biology
- Production of poly-3-hydroxybutyrate from mixed culture
- Cellular and Molecular Biology
- Molecular and immunological characterization of Lp1, the 34 kDa outer membrane lipoprotein of Pseudomonas aeruginosa
- Cellular and Molecular Biology
- Antibacterial activity of a microemulsion loaded with cephalosporin
- Cellular and Molecular Biology
- Isolation and characterization of Vibrio alginolyticus from cultured amphioxus Branchiostoma belcheri tsingtauense
- Cellular and Molecular Biology
- Somatic embryogenesis in Phlox paniculata – histological analysis
- Botany
- Overexpression of the novel Zygophyllum xanthoxylum C2H2-type zinc finger gene ZxZF improves drought tolerance in transgenic Arabidopsis and poplar
- Botany
- Vegetation diversity of the two Dinaric karstic rivers in Bosnia and Herzegovina
- Botany
- Diversity and spatial patterns of plant communities in volcanic temporary ponds of Sicily (Italy)
- Zoology
- Erythraeus (Erythraeus) pistacicus sp. n. (Trombidiformes: Erythraeidae) from southern Iran, and notes on other Erythraeus spp.
- Zoology
- Studies on eucrenal-hypocrenal zonation of springs along the river mainstream: A case study of a karst canyon in Bosnia and Herzegovina
- Zoology
- Different catching series from light and pheromone trapping of Helicoverpa armigera (Lepidoptera: Noctuidae)
- Zoology
- Newts in the nature reserve Dönche: Evaluation of diversity, breeding behaviour and threats due to a decline in habitat suitability
- Zoology
- Antioxidant status of rabbits after treatment with epicatechin and patulin