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Viability During Storage and Stability of Plasmids During Storage and Subculturing in Strains of Vibrio anguillarum

Acta Veterinaria Scandinavica volume 38, pages 315–322 (1997)Cite this article

Abstract

The stability of plasmids, 3.3 kb–~200 kb, in 8 strains of Vibrio anguillarum displaying different plasmid profiles and reactions with O-antisera was investigated over an 18 months period. All plasmid profiles proved to be resistant to storage at different temperatures but strains stored at 37 °C were only viable for a short period of maximum 2 months. Strains stored at 5°C and 20°C were viable for a longer period. Viable strains maintained their plasmid profile throughout the experiment, except in 2 cases where a 67 kb and a 200 kb plasmid were lost. Strains stored at -80 °C all remained viable and maintained their plasmid profile throughout the study. By subcultivating daily for up to 100 successive days, most strains maintained their plasmid profiles. Only 2 strains lost their plasmid. When picking 100 single colonies from agar plates, none of the colonies showed plasmid profiles deviating from the expected. The results suggest that plasmid profiles among V anguillarium are very stable during subculturing, storage and laboratory handling using standard laboratory procedures, and thus, reliable for epidemiological investigations. In a second experiment, 2 pairs of 2 strains were grown together in mixed cultures. They were identical in all traits, except that one strain in each pair harboured the 67 kb pJMl-like virulence plasmid, whereas the other had lost this plasmid. The result showed that the growth rate was the same for strains with and without the plasmid, indicating that under laboratory conditions, this plasmid is neither a benefit nor the opposite for bacterial growth.

References

  • Aoki T, Egusa S, Arai T: Detection of R factors in naturally occurring Vibrio anguillarum strains. Anti-microb. Agents Chemother. 1974, 6, 534–538.

    Article  CAS  Google Scholar 

  • Aoki T, Kanasawa T, Kitao T: Epidemiological surveillance of drug resistant Vibrio anguillarum strains. Fish Pathol. 1985,29, 199–208.

    Article  Google Scholar 

  • Austin B, Alsina M, Austin DA, Blanch A, Grimont F, Grimont PAD, Jofre J, Koblavi S, Larsen JL, Pedersen K, Tiainen T, Verdonck L, Swings J: Identification and typing of Vibrio anguillarum: a comparison of different methods. System. Appl. Microbiol. 1995, 18, 285–302.

    Article  Google Scholar 

  • Austin B, Austin DA: Bacterial Fish Pathogens–Disease of Farmed and Wild Fish, 2nd edn. Ellis Horwood. Chichester, England, 1993.

    Google Scholar 

  • Caldwell BA, Ye C, Griffith RP, Moyer CL, Monta RY: Plasmid expression and maintenance during long-term starvation-survival of bacteria in well water. Appl. Environ. Microbiol. 1989, 55, 1860–1864.

    Article  CAS  Google Scholar 

  • Crosa JH: A plasmid associated with virulence in the marine fish pathogen Vibrio anguillarum specifies an iron-sequestering system. Nature 1980, 284, 566–568.

    Article  CAS  Google Scholar 

  • Crosa JH, Hodges LL, Schiewe MH: Curing of a plasmid is correlated with an attenuation of virulence in the marine fish pathogen Vibrio anguillarum. Infect. Immun. 1980, 27, 897–902.

    Article  CAS  Google Scholar 

  • Hoyt PR, Sizemore RK: Competetive dominance in a bacteriocin-producing Vibrio harveyi strain. Appl. Environ. Microbiol. 1982, 44, 53–658.

    Article  Google Scholar 

  • Kado CI, Liu ST: Rapid procedure for detection and isolation of large and small plasmids. J. Bacteriol. 1981,745, 1365–1373.

    Article  Google Scholar 

  • Larsen JL: Vibrio anguillarum: characterization, ecology, and pathogenicity. Dissertation. DSR Press, Copenhagen, Denmark, 1990.

    Google Scholar 

  • Larsen JL, Olsen JE: Occurrence of plasmids in Danish isolates of Vibrio anguillarum serovars Ol and 02 and association with phenotypic characteristics. Appl. Environ. Microbiol. 1991, 57, 2158–2163.

    Article  CAS  Google Scholar 

  • Larsen JL, Pedersen K, Dalsgaard I: Vibrio anguillarum serovars associated with vibriosis in fish. J. Fish Dis. 1994,77,259–267.

    Article  Google Scholar 

  • Olsen JE, Brown DJ, Baggesen DL, Bisgaard M: Stability of plasmids in five strains of Salmonella maintained in stab culture at different temperatures. J. Appl. Bacteriol. 1994, 77, 155–159.

    Article  CAS  Google Scholar 

  • Olsen JE, Larsen JL: Ribotypes and plasmid contents of Vibrio anguillarum strains in relation to serovar. Appl. Environ. Microbiol. 1993, 59, 3863–3870.

    Article  CAS  Google Scholar 

  • Pedersen K, Dalsgaard I, Larsen JL: Characterization of atypical Aeromonas salmonicida isolates by ribotyping and plasmid profiling. J. Appl. Bacteriol. 1996a, 80, 37–44.

    Article  CAS  Google Scholar 

  • Pedersen K, Dalsgaard I, Larsen JL: Vibrio damsela associated with disease in fish in Denmark. Appl. Environ. Microbiol. 1997a, 63, 3711–3715.

    Article  CAS  Google Scholar 

  • Pedersen K, Gram L, Austin DA, Austin B: Pathogenicity of Vibrio anguillarum serogroup 01 strains compared to plasmids, outer membrane protein profiles and siderophore production. J. Appl. Microbiol. 1997b, 82, 365–371.

    Article  CAS  Google Scholar 

  • Pedersen K, Grisez L, van Hout R, Tiainen T, Ollevier F, Larsen JL: Revised and extended O-serotyping scheeme for Vibrio anguillarum. 1998 (submitted).

    Google Scholar 

  • Pedersen K, Larsen JL: Evidence for the existence of distinct populations of Vibrio anguillarum serogroup 01 based on plasmid profiles and ribotyping. Appl. Environ. Microbiol 1995, 61, 2292–2296.

    Article  CAS  Google Scholar 

  • Pedersen K, Tiainen T, Larsen JL: Plasmid profiles, restriction fragment length polymorphisms and O-serotypes among Vibrio anguillarum isolates. Epidemiol. Infect. 1996b, 777, 471–478.

    Article  Google Scholar 

  • Sarra PG, Vescovo M, Bottazzi V: Antagonism and adhesion among isogenic strains of Lactobacillus reuteri in the caecum of gnotobiotic mice. Micro-biologica 1989, 12, 69–74.

    CAS  Google Scholar 

  • Skov MN, Pedersen K, Larsen JL: Comparison of ribotyping, pulsed- field gel electrophoresis and plasmid profiling for typing of Vibrio anguillarum serovar 01. Appl. Environ. Microbiol. 1995, 61, 1540–1545.

    Article  CAS  Google Scholar 

  • Summers DK: The biology of plasmids. Blackwell Science Ltd., Oxford, UK, 1996.

    Book  Google Scholar 

  • Sorensen UBS, Larsen JL: Serotyping of Vibrio anguillarum. Appl. Environ. Microbiol. 1986, 51, 593–597.

    Article  CAS  Google Scholar 

  • Tiainen T, Pedersen K, Larsen JL: Ribotyping and plasmid profiling of Vibrio anguillarum serovar 02 and Vibrio ordalii. J. Appl. Bacteriol. 1995, 79, 384–392.

    Article  CAS  Google Scholar 

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Acknowledgment

The technical assistance of Ms. Farah Bahrani, Ms. Annette Rasmussen, and Mrs. Dianne F. Rasmussen, and the financial support of The Danish Agricultural and Veterinary Research Council, grant no. 9503658, and the European Union, grant no. AIR-CT92-0341, are gratefully appreciated.

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  1. Laboratory of Fish Diseases, Department of Veterinary Microbiology, The Royal Veterinary and Agricultural University, Biilowsvej 13, DK-1870, Fredenksberg C, Denmark

    K. Pedersen

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Pedersen, K. Viability During Storage and Stability of Plasmids During Storage and Subculturing in Strains of Vibrio anguillarum. Acta Vet Scand 38, 315–322 (1997). https://doi.org/10.1186/BF03548477

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Acta Veterinaria Scandinavica

ISSN: 1751-0147