Archives of Microbiology

, Volume 191, Issue 6, pp 493–500 | Cite as

Enzymatic, outer membrane proteins and plasmid alterations of starved Vibrio parahaemolyticus and Vibrio alginolyticus cells in seawater

  • Fethi Ben Abdallah
  • Héla Kallel
  • Amina Bakhrouf
Original Paper


The marine bacteria Vibrio parahaemolyticus and V. alginolyticus were incubated in seawater for 8 months to evaluate their adaptative responses to starvation. The starved cells showed an altered biochemical and enzymatic profiles, respectively, on Api 20E and Api ZYM systems and an evolution to the filterable minicells state capable to pass membrane pore size 0.45 μm. Outer membrane proteins patterns of stressed bacteria were also altered. Indeed, these modifications were manifested by the appearance and/or disappearance of bands as well as in the level of expression of certain proteins. Plasmids profiles analysis showed that V. alginolyticus ATCC 33787 lost three plasmids, whereas other tested strains conserved their initial profiles.


Vibrio Seawater Starvation Enzymatic Morphological Outer membrane proteins Plasmids Alterations 



We are grateful to M’SELMI Abdelhamid principal technical; USCR: Unité des Services Communs de Recherche, Faculté des Sciences de Tunis. Campus Universitaire. Tunisia.


  1. Bakhrouf A, Jeddi M, Bouddabous A, Gauthier MJ (1989) Evolution of Pseudomonas aeruginosa cells towards a filterable stage in seawater. FEMS Microbiol Lett 50:187–190PubMedCrossRefGoogle Scholar
  2. Bakhrouf A, Jeddi M, Gauthier MJ (1992) Modifications des caractères culturaux et biochimiques du Salmonella paratyphi B après incubation dans l’eau de mer. Can J Microbiol 55:871–874Google Scholar
  3. Bakhrouf-Ben Fedhila A, Jeddi M, Bouddabous A, Gauthier MJ (1990) Production of filterable minicells by Salmonella paratyphi B in seawater. Microbiol Lett 43:123–129Google Scholar
  4. Ben Abdallah F, Chaieb K, Snoussi M, Bakhrouf A, Gaddour K (2007) Phenotypic variations and molecular identification of Salmonella enterica serovar Typhimurium cells under starvation in seawater. Curr Microbiol 55:485–491PubMedCrossRefGoogle Scholar
  5. Ben Abdallah F, Chaieb K, Kallel H, Bakhrouf A (2009) RT-PCR assays for in vivo expression of Vibrio alginolyticus virulence genes in cultured gilthead Dicentrarchus labrax and Sparus aurata. Ann Microbiol 59:1–5Google Scholar
  6. Ben Kahla NA, Besbes A, Chaieb K, Rouabhia M, Bakhrouf A (2007) Survival of Vibrio alginolyticus in seawater and retention of virulence of its starved cells. Mar Environ Res 64:469–478CrossRefGoogle Scholar
  7. Ben Kahla NA, Besbes A, Bakhrouf A (2008) Survival of Vibrio fluvialis in seawater under starvation conditions. Microbiol Res 163:323–328CrossRefGoogle Scholar
  8. Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523PubMedCrossRefGoogle Scholar
  9. Cai SJ, Inouye M (2002) EnvZ–OmpR interaction and osmoregulation in Escherichia coli. J Biol Chem 277:24155–24161PubMedCrossRefGoogle Scholar
  10. Carroll JA, Garon CF, Schwan TG (1999) Effects of environmental pH on membrane proteins in Borrelia burgdorferi. Infect Immun 67:3181–3187PubMedGoogle Scholar
  11. Chen HY, Livermore DM (1993) Activity of cefepime and other beta-lactam antibiotics against permeability mutants of Escherichia coli and Klebsiella pneumoniae. J Antimicrob Chemother 32(Suppl B):63–74PubMedGoogle Scholar
  12. Contreras I, Muñoz L, Toro CS, Mora GC (1995) Heterologous expression of Escherichia coli porin genes in Salmonella typhi Ty2: regulation by medium osmolarity, temperature and oxygen availability. FEMS Microbiol Lett 133:105–111PubMedCrossRefGoogle Scholar
  13. Costerton JW, Cheng K (1975) The role of the bacterial cell envelope in antibiotic resistance. J Antimicrob Chemother 1:363–377PubMedCrossRefGoogle Scholar
  14. Cullen PA, Cordwell SJ, Bulach DM, Haake DA, Adler B (2002) Global analysis of outer membrane proteins from Leptospira interrogans serovar Lai. Infect Immun 70:2311–2318PubMedCrossRefGoogle Scholar
  15. Dedieu L, Pages JM, Bolla JM (2002) Environmental regulation of Campylobacter jejuni major outer membrane protein porin expression in Escherichia coli monitored by using green fluorescent protein. Appl Environ Microbiol 68:4209–4215PubMedCrossRefGoogle Scholar
  16. Denner EBM, Vybiral D, Fischer UR, Velimirov B, Busse HJ (2002) Vibrio calviensis sp. nov., a halophilic, facultatively oligotrophic 0.2 μm-filterable marine bacterium. Int J Syst Bacteriol 52:549–553Google Scholar
  17. Feodorova VA, Devdariani ZL (2001) Expression of acid-stable proteins and modified lipopolysaccharide of Yersinia pestis in acidic growth medium. J Med Microbiol 50:979–985PubMedGoogle Scholar
  18. Gauthier MJ, Munro PM, Breittmayer VA (1988) Damage to surface colonisation factors of enters adhesive Escherichia coli for starvation in seawater. Microbios Lett 38:37–45Google Scholar
  19. Hernandez-Alles S, Albert S, Alvarez D, Domenech-Sanchez A, Martinez-Martinez L, Gil J, Tomas JM, Bendi VJ (1999) Porin expression in clinical isolates of Klebsiella pneumoniae. Microbiology 145:673–679PubMedCrossRefGoogle Scholar
  20. Hwang PM, Choy WY, Lo EI, Chen L, Forman-Kay JD, Raetz CR, Privé GG, Bishop RE, Kay LE (2002) Solution structure and dynamics of the outer membrane enzyme PagP by NMR. Proc Natl Acad Sci USA 99:13560–13565PubMedCrossRefGoogle Scholar
  21. Jiang X, Chai T (1996) Survival of Vibrio parahaemolyticus at low temperatures under starvation conditions and subsequent resuscitation of viable, nonculturable cells. Appl Environ Microb 62:1300–1305Google Scholar
  22. Kjelleberg S (1993) Starvation in bacteria. Plenum, New YorkGoogle Scholar
  23. Kjelleberg S, Hermannson M, Marden P, Jones GW (1987) The transient phase between growth and nongrowth of heterotrophic bacteria, with emphasis on the marine environment. Annu Rev Microbiol 41:25–49PubMedCrossRefGoogle Scholar
  24. Kustos I, Kocsis B, Kilár F (2007) Bacterial outer membrane protein analysis by electrophoresis and microchip technology. Expert Rev Proteomics 4:91–106PubMedCrossRefGoogle Scholar
  25. Laemmli UK (1970) Cleavage of structural proteins for the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  26. Leal-Balbino TC, Leal NC, Lopes CV, de Almeida AMP (2004) Differences in the stability of the plasmids of Yersinia pestis cultures in vitro: Impact on virulence. Mem Inst Oswaldo Cruz 99:727–732PubMedCrossRefGoogle Scholar
  27. Liu PV (1957) Survey of hemolysin production among species of Pseudomonas. J Bacteriol 74:718–727PubMedGoogle Scholar
  28. Martinez MB, Flickinger M, Higgins L, Krick T, Nelsestuen GL (2001) Reduced outer membrane permeability of Escherichia coli O157:H7: suggested role of modified outer membrane porins and theoretical function in resistance to antimicrobial agents. Biochemistry 40:11965–11974PubMedCrossRefGoogle Scholar
  29. Molloy MP, Herbert BR, Slade MB, Rabilloud T, Nouwens AS, Williams KL, Gooley AA (2000) Proteomic analysis of the Escherichia coli outer membrane. Eur J Biochem 267:2871–2881PubMedCrossRefGoogle Scholar
  30. Morita RY (1997) Bacteria in oligotrophic environments: starvation-survival lifestyle. Chapman & Hall, New YorkGoogle Scholar
  31. Mushtaq S, Ge Y, Livermore DM (2004) Doripenem versus Pseudomonas aeruginosa in vitro: activity against characterized isolates, mutants, and transconjugants and resistance selection potential. Antimicrob Agents Chemother 8:3086–3092CrossRefGoogle Scholar
  32. Obonyo M, Munderloh UG, Sam TN, Kurtti TJ (2002) Cultivation at 37 degrees C enhances Borrelia burgdorferi sensu stricto infectivity for hamsters. Med Microbiol Immunol 191:33–39PubMedCrossRefGoogle Scholar
  33. Provenzano D, Lauriano CM, Klose KE (2001) Characterization of the role of the ToxR-modulated outer membrane porins OmpU and OmpT in Vibrio cholerae virulence. J Bacteriol 183:3652–3662PubMedCrossRefGoogle Scholar
  34. Puig M, Fusté C, Viñas M (1993) Outer membrane proteins from Serratia marcescens. Can J Microbiol 39:108–111PubMedCrossRefGoogle Scholar
  35. Rahmati-Bahram A, Magee JT, Jackson SK (1996) Temperature-dependent aminoglycoside resistance in Stenotrophomonas (Xanthomonas) maltophilia; alterations in protein and lipopolysaccharide with growth temperature. J Antimicrob Chemother 37:665–676PubMedCrossRefGoogle Scholar
  36. Rosen R, Buttner K, Schmid R, Hecker M, Ron EZ (2001) Stress-induced proteins of Agrobacterium tumefaciens. FEMS Microbiol Ecol 35:277–285PubMedCrossRefGoogle Scholar
  37. Sabri MY, Zamri-Saad M, Mutalib AR, Israf DA, Muniandy N (2000) Efficacy of an outer membrane protein of Pasteurella haemolytica A2, A7 or A9-enriched vaccine against intratracheal challenge exposure in sheep. Vet Microbiol 73:13–23PubMedCrossRefGoogle Scholar
  38. Sambrook PL, Peterson BC, Gerding DN, Cleary PP (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
  39. Shand GH, Anwar H, Brown MR (1988) Outer membrane proteins of polymyxin resistant Pseudomonas aeruginosa: effect of magnesium depletion. J Antimicrob Chemother 22:811–821PubMedCrossRefGoogle Scholar
  40. Siegele DA, Kolter R (1992) Life after log. J Bacteriol 174:345–348PubMedGoogle Scholar
  41. Srinivasan S, Kjellberg S (1998) Cycles of famine and feast: the starvation and outgrowth strategies of a marine Vibrio. J Biosci 23:501–511CrossRefGoogle Scholar
  42. Sunde M, Sorum H (2001) Self-transmissible multidrug resistance plasmids in Escherichia coli of the normal intestinal flora of healthy swine. Microbial Drug Resistance 7:191–196PubMedCrossRefGoogle Scholar
  43. Tabor PS, Ohwada K, Colwell RR (1981) Filterable marine bacteria found in the deep sea distribution, taxonomy, and response to starvation. Microb Ecol 7:67–83CrossRefGoogle Scholar
  44. Van Gemerden H, Kuen JG (1984) Strategies for growth and evolution of microorganisms in oligotrophic habitats. In: Hobbie JE, Williams PJB (eds) Heterotrophic activity in the sea. Nato Conference series IV. Plenum Press, New York, pp 25–54Google Scholar
  45. Vybiral D, Denner EBM, Haller CM, Busse HJ, Witte A, Hofle MG, Velimirov B (1999) Polyphasic classification of 0.2 μm filterable bacteria from the Western Mediterranean Sea. Syst Appl Microbiol 22:635–646PubMedGoogle Scholar
  46. Wibbenmeyer JA, Provenzano D, Landry CF, Klose KE, Delcour AH (2002) Vibrio cholerae OmpU and OmpT Porins are differentially affected by bile. Infect Immun 70:121–126PubMedCrossRefGoogle Scholar
  47. Wu L, Lin X, Wang F, Ye D, Xiao X, Wang S, Peng X (2006) OmpW and OmpV are required for NaCl regulation in Photobacterium damsela. J Proteome Res 5:2250–2257PubMedCrossRefGoogle Scholar
  48. Xu C, Ren H, Wang S, Peng X (2004) Proteomic analysis of salt-sensitive outer membrane proteins of Vibrio parahaemolyticus. Res Microbiol 155:835–842PubMedCrossRefGoogle Scholar
  49. Xu C, Wang S, Ren H, Lin X, Wu L, Peng X (2005) Proteomic analysis on the expression of outer membrane proteins of Vibrio alginolyticus at different sodium concentrations. Proteomics 5:3142–3152PubMedCrossRefGoogle Scholar
  50. Yoshida T, Qin L, Inouye M (2002) Formation of the stoichiometric complex of EnvZ, a histidine kinase, with its response regulator, OmpR. Mol Microbiol 46:1273–1282PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Fethi Ben Abdallah
    • 1
    • 2
  • Héla Kallel
    • 2
  • Amina Bakhrouf
    • 1
  1. 1.Laboratoire d’Analyse, Traitement et Valorisation des Polluants de l’Environnement et des ProduitsFaculté de Pharmacie Rue AvicenneMonastirTunisia
  2. 2.Unité de Fermentation et de Développement de Vaccins VirologiquesInstitut Pasteur de TunisTunisTunisia

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