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Volume 154, Issue 4

A mutant in one of two loci of a TonB system in shows attenuated virulence and confers protection against cold water disease Free

Abstract

is a psychrotrophic fish-pathogenic bacterium that causes cold water disease (CWD) in salmonids. By means of Tn mutagenesis a mutant named FP1033, deficient in growth on iron-depleted medium, was previously isolated. FP1033 recovered the parental phenotype in the presence of iron. The gene disrupted by the transposon in this mutant encoded a protein with similarity to ExbD proteins, which are members of the TonB complex system involved in iron uptake mediated by siderophores. Analysis of the DNA surrounding the transposon insertion showed the presence of a cluster of genes composed of , two ( and ) and loci. RT-PCR analysis and complementation studies indicated that these genes are transcribed as an operon and that the  : : Tn phenotype was caused by the lack of ExbD2. FP1033 showed decreased virulence and conferred a high level of protection in rainbow trout fry after vaccination. This is believed to be the first report of a attenuated strain that induces a protective immune response in rainbow trout against CWD. These results suggest that the locus from this particular TonB system is a suitable target to generate a live attenuated vaccine.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): CWD, cold water disease and RPS, relative percentage survival
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2008-04-01
2024-04-20
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References

  1. Agarwal S., Hunnicutt D. W., McBride M. J. 1997; Cloning and characterization of the Flavobacterium johnsoniae ( Cytophaga johnsonae) gliding motility gene, gldA. Proc Natl Acad Sci U S A 94:12139–12144
     [Google Scholar]
  2. Álvarez B., Secades P., McBride M. J., Guijarro J. A. 2004; Development of genetic techniques for the psychrotrophic fish pathogen Flavobacterium psychrophilum. Appl Environ Microbiol 70:581–587
     [Google Scholar]
  3. Álvarez B., Secades P., Prieto M., McBride M. J., Guijarro J. A. 2006; A mutation in Flavobacterium psychrophilum tlpB inhibits gliding motility and induces biofilm formation. Appl Environ Microbiol 72:4044–4053
     [Google Scholar]
  4. Andrews S. C., Robinson A. K., Rodriguez-Quiñones F. 2003; Bacterial iron homeostasis. FEMS Microbiol Rev 27:215–237
     [Google Scholar]
  5. Aoki M., Kondo M., Nakatsuka Y., Kawai K., Oshima S.-I. 2007; Stationary phase culture supernatant containing membrane vesicles induced immunity to rainbow trout Oncorhynchus mykiis fry syndrome. Vaccine 25:561–569
     [Google Scholar]
  6. Beare P. A., For J., Martin L. W., Lamont I. L. 2003; Siderophore-mediated cell signalling in Pseudomonas aeruginosa: divergent pathways regulate virulence factor production and siderophore receptor synthesis. Mol Microbiol 47:195–207
     [Google Scholar]
  7. Braun V. 1995; Energy-coupled transport and signal transduction through the gram-negative outer membrane via tonB-exbB-exbD-dependent receptor proteins. FEMS Microbiol Rev 16:295–307
     [Google Scholar]
  8. Braun V., Braun M. 2002; Active transport of iron and siderophore antibiotics. Curr Opin Microbiol 5:194–201
     [Google Scholar]
  9. Bruun M. S., Schimdt A. S., Madsen L., Dalsgaard I. 2000; Antimicrobial resistance patterns in Danish isolates of Flavobacterium psychrophilum. Aquaculture 187:201–212
     [Google Scholar]
  10. Cabello F. C. 2006; Heavy use of prophylactic antibiotics in aquaculture: a growing problem for human and animal health and for the environment. Environ Microbiol 8:1137–1144
     [Google Scholar]
  11. del Cerro A., Mendoza M. C., Guijarro J. A. 2002; Usefulness of a TaqMan-based polymerase chain reaction assay for the detection of the fish pathogen Flavobacterium psychrophilum. J Appl Microbiol 93:149–156
     [Google Scholar]
  12. Dumetz F., Duchaud E., LaPatra S. E., Le Marrec C., Cleverol S., Urdaci M.-C., Le Hénaff M. 2006; A protective immune response is generated in rainbow trout by an OmpH-like surface antigen (P18) of Flavobacterium psychrophilum. Appl Environ Microbiol 72:4845–4852
     [Google Scholar]
  13. Enard C., Expert D. 2000; Characterization of a tonB mutation in Erwinia chrysanthemi 3937: TonB is a member of the enterobacterial TonB family. Microbiology 146:2051–2058
     [Google Scholar]
  14. Fernández L., Márquez I., Guijarro J. A. 2004; Identification of specific in vivo-induced ( ivi) genes in Yersinia ruckeri and analysis of ruckerbactin, a catecholate siderophore iron acquisition system. Appl Environ Microbiol 70:5199–5207
     [Google Scholar]
  15. Garcia C., Pozet F., Michel C. 2000; Standardization of experimental infection with Flavobacterium psychrophilum, the agent of rainbow trout Onchorhynchus mykiss fry syndrome. Dis Aquat Organ 42:191–197
     [Google Scholar]
  16. Hernanz Moral C., Flaño del Castillo E., López Fierro P., Villena Cortés A., Anguita Castillo J., Cascón Soriano A., Sánchez Salazar M., Razquín Peralta B., Naharro Carrasco G. 1998; Molecular characterization of the Aeromonas hydrophila aroA gene and potential use of an auxotrophic aroA mutant as a live attenuated vaccine. Infect Immun 66:1813–1821
     [Google Scholar]
  17. Hunnicutt D. W., McBride M. J. 2000; Cloning and characterization of the Flavobacterium johnsoniae gliding-motility genes gldB and gldC. J Bacteriol 182:911–918
     [Google Scholar]
  18. Izumi S., Aranishi F. 2004; Relationship between gyrA mutations and quinolone resistance in Flavobacterium psychrophilum isolates. Appl Environ Microbiol 70:3968–3972
     [Google Scholar]
  19. Kondo M., Kawai K., Obake M., Nakano N., Oshima S. 2003; Efficacy of oral vaccine against bacterial coldwater disease in ayu Plecoglossus altivelis. Dis Aquat Organ 55:261–264
     [Google Scholar]
  20. Köster W. 2001; ABC transporter-mediated uptake of iron, siderophores, heme and vitamin B12. Res Microbiol 152:291–301
     [Google Scholar]
  21. LaFrentz B. R., LaPatra S. E., Jones G. R., Cain K. D. 2004; Protective immunity in rainbow trout Oncorhynchus mykiis following immunization with distinct molecular mass fractions isolated from Flavobacterium psychrophilum. Dis Aquat Organ 59:17–26
     [Google Scholar]
  22. Madetoja J., Lönnström L.-G., Björkblom C., Uluköy G., Bylund G., Syvertsen C., Gravningen K., Norderhus E.-A., Wiklund T. 2006; Efficacy of injection vaccines against Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiis (Walbaum. J Fish Dis 29:9–20
     [Google Scholar]
  23. Maunsell B., Adams C., O'Gara F. 2006; Complex regulation of AprA metalloprotease in Pseudomonas fluorescens M114: evidence for the involvement of iron, the ECF sigma factor, PbrA and pseudobactin M114 siderophore. Microbiology 152:29–42
     [Google Scholar]
  24. McBride M. J., Braun T. F., Brust J. L. 2003; Flavobacterium johnsoniae GldH is a lipoprotein that is required for gliding motility and chitin utilization. J Bacteriol 185:6648–6657
     [Google Scholar]
  25. Michel C., Antonio D., Hedrick R. P. 1999; Production of viable cultures of Flavobacterium psychrophilum approach and control. Res Microbiol 150:351–358
     [Google Scholar]
  26. Møller J. D., Ellis A. E., Barnes A. C., Dalsgaard I. 2005; Iron acquisition mechanisms of Flavobacterium psychrophilum. J Fish Dis 28:391–398
     [Google Scholar]
  27. Nematollahi A., Decostere A., Pasmas F., Haesebrouck F. 2003; Flavobacterium psychrophilum infections in salmonid fish. J Fish Dis 26:563–574
     [Google Scholar]
  28. Norqvist A., Hagström A., Wolf-Watz H. 1989; Protection of rainbow trout against vibriosis and furunculosis by the use of attenuated strains of Vibrio anguillarum. Appl Environ Microbiol 55:1400–1405
     [Google Scholar]
  29. Payne S. M. 1993; Iron acquisition in microbial pathogenesis. Trends Microbiol 1:66–69
     [Google Scholar]
  30. Payne S. M., Wyckoff E. E., Murphy E. R., Oglesby A. G., Boulette M. L., Davies N. M. 2006; Iron and pathogenesis of Shigella: iron acquisition in intracellular environment. Biometals 19:173–180
     [Google Scholar]
  31. Postle K. 1990; TonB and gram-negative dilemma. Mol Microbiol 4:2019–2025
     [Google Scholar]
  32. Pradel E., Guiso N., Menozzi F. D., Locht C. 2000; Bordetella pertussis TonB, a Bvg-independent virulence determinant. Infect Immun 68:1919–1927
     [Google Scholar]
  33. Rahman H., Kuroda A., Dijkstra J. M., Kiryu I., Nakanishi T., Ototake M. 2002; The outer membrane fraction of Flavobacterium psychrophilum induces protective immunity in rainbow trout and ayu. Fish Shellfish Immunol 12:169–179
     [Google Scholar]
  34. Reed L. J., Muench H. 1938; A simple method of estimating fifty percent endpoints. Am J Hyg 27:493–497
     [Google Scholar]
  35. Reeves S. A., Torres A. G., Payne S. M. 2000; TonB is required for intracellular growth and virulence of Shigella dysenteriae. Infect Immun 68:6329–6336
     [Google Scholar]
  36. Schmidt A. S., Bruun M. S., Dalsgaard I., Pedersen K., Larsen J. L. 2000; Occurrence of antimicrobial resistance in fish-pathogenic and environmental bacteria associated with four Danish rainbow trout farms. Appl Environ Microbiol 66:4908–4915
     [Google Scholar]
  37. Schwyn B., Neilands J. B. 1987; Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56
     [Google Scholar]
  38. Seliger S. S., Mey A. R., Valle A. M., Payne S. M. 2001; The two TonB systems of Vibrio cholerae: redundant and specific functions. Mol Microbiol 39:801–812
     [Google Scholar]
  39. Simon R., Priefer U., Puhler A. 1983; A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Bio/technology 1:784–791
     [Google Scholar]
  40. Smith C. J., Rogers M. B., McKee M. L. 1992; Heterologous gene expression in Bacteroides fragilis. Plasmid 27:141–154
     [Google Scholar]
  41. Stork M., Di Lorenzo M., Mouriño S., Osorio C. R., Lemos M. L., Crosa J. H. 2004; Two TonB systems function in iron transport in Vibrio anguillarum, but only one is essential for virulence. Infect Immun 72:7326–7329
     [Google Scholar]
  42. Temprano A., Riano J., Yugueros J., González P., De Castro L., Villena A., Luengo J. M., Naharro G. 2005; Potential use of a Yersinia ruckeri O1 auxotrophic aroA mutant as a live attenuated vaccine. J Fish Dis 28:419–427
     [Google Scholar]
  43. Tokuda M., Chem W., Karunakaran T., Kuramitsu H. K. 1998; Regulation of protease expression in Porphyromonas gingivalis. Infect Immun 66:5232–5237
     [Google Scholar]
  44. Torres A. G., Redford P., Welch R. A., Payne S. M. 2001; TonB-dependent systems of uropathogenic Escherichia coli: aerobactin and heme transport and TonB are required for virulence in the mouse. Infect Immun 69:6179–6185
     [Google Scholar]
  45. Vaughan L. M., Smith P. R., Foster T. J. 1993; An aromatic-dependent mutant of the fish pathogen Aeromonas salmonicida is attenuated in fish and is effective as a live vaccine against the salmonid disease furunculosis. Infect Immun 61:2172–2181
     [Google Scholar]
  46. Wandersman C., Delepelaire P. 2004; Bacterial iron sources: from siderophores to hemophores. Annu Rev Microbiol 58:611–647
     [Google Scholar]
  47. Wiggerich H.-G., Pühler A. 2000; The exbd2 gene as well as the iron-uptake genes tonB, exbB and exbD1 of Xanthomonas campestris pv. campestris are essential for the induction of a hypersensitive response on pepper ( Capsicum annuum). Microbiology 146:1053–1060
     [Google Scholar]
  48. Wooldridge K. G., Williams P. H. 1993; Iron uptake mechanisms of pathogenic bacteria. FEMS Microbiol Rev 12:325–348
     [Google Scholar]
  49. Wyckoff E. E., May A. R., Payne S. M. 2007; Iron acquisition in Vibrio cholerae. Biometals 20:405–416
     [Google Scholar]
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A mutant in one of two exbD loci of a TonB system in Flavobacterium psychrophilum shows attenuated virulence and confers protection against cold water disease
Microbiology 154, 1144 (2008); https://doi.org/10.1099/mic.0.2007/010900-0
/content/journal/micro/10.1099/mic.0.2007/010900-0
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