1887

Microbiology Society logo

Volume 146, Issue 1

Invasion of fish epithelial cells by subsp. : evidence for receptor specificity, and effect of capsule and serum Free

Abstract

subsp. is a fish pathogen which causes serious disease in commercial warmwater fish species. Because information on the initial stages of the infection is scarce, an investigation of the invasion ability of this pathogen was undertaken utilizing a fish epithelial cell line (epithelioma papillosum carpio, EPC), a virulent capsulated strain of (MT1415), an avirulent non-capsulated strain of (EPOY- 8803-II) and HB101 as a non-invasive control. was found to be able to adhere to and invade fish epithelial cells and remain inside them for 6–9 h. There were no significant differences in invasiveness between the capsulated and non-capsulated strains. A kinetics study demonstrated that invasiveness was more efficient at low m.o.i., reaching saturation at higher m.o.i., suggesting internalization may be receptor-mediated. Invasion efficiency (IE) was significantly higher than in the control HB101. Engulfment of bacteria was possibly by an endocytic process and was unaffected by killing the bacteria with UV light. However, heat-killed bacteria had significantly reduced invasion capability. Ultrastructural studies showed that inside the epithelial cells, the bacteria remained within large vacuoles for a few hours and no evidence of intracellular replication was found, by either fluorescence or electron microscopic studies. Normal sea bass serum slightly reduced the invasion capability of the MT1415 strain, but heat-inactivated normal serum had no effect. On the other hand, heat-inactivated fish antiserum raised against the same strain reduced the percentage of invaded epithelial cells by 50%. As for other pathogens, an intracellular phase of may be a mechanism to delay or avoid phagocytosis and host immune responses, favouring the spread of infection.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): EPC, epithelioma papillosum carpio , fish epithelial cells , IE, invasion efficiency , invasion efficiency and Photobacterium damselae subsp. piscicida
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-1-21
2000-01-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/1/1460021a.html?itemId=/content/journal/micro/10.1099/00221287-146-1-21&mimeType=html&fmt=ahah

References

  1. Amaro C., Biosca E. G., Fouz B., Toranzo A. E., Garay E. 1994; Role of iron, capsule, and toxins in the pathogenicity of Vibrio vulnificus biotype 2 for mice. Infect Immun 62:759–763
     [Google Scholar]
  2. Arijo S., Borrego J. J., Zorrilla I., Balebona M. C., Moriñigo M. A. 1998; Comparison of the immune response of gilt-head seabream (Sparus aurata L.) to capsulated and uncapsulated strains of Photobacterium damsela subsp. piscicida. Fish Shellfish Immunol 8:63–72 [CrossRef]
     [Google Scholar]
  3. Bandı́n I., Rivas C., Santos Y., Secombes C. J., Barja J. L., Ellis A. E. 1995; Effect of serum factors on the survival of Renibacterium salmoninarum within rainbow trout macrophages. Dis Aquat Org 23:221–227 [CrossRef]
     [Google Scholar]
  4. Barnes A. C., Lewin C. S., Hastings T. S., Amyes S. G. B. 1991; Susceptibility of the fish pathogen Aeromonas salmonicida to flumequine. Antimicrob Agents Chemother 35:2634–2635 [CrossRef]
     [Google Scholar]
  5. Barnes A. C., Balebona M. C., Horne M. T., Ellis A. E. 1999; Superoxide dismutase and catalase in Photobacterium damselae subsp. piscicida and their roles in resistance to reactive oxygen species. Microbiology 145:483–494 [CrossRef]
     [Google Scholar]
  6. Chan R., Acres S. D., Costerton J. W. 1982; Use of specific antibody to demonstrate glycocalyx, K99 pili, and the spatial relationships of K99+ enterotoxigenic Escherichia coli in the ileum of colostrum-fed calves. Infect Immun 37:1170–1180
     [Google Scholar]
  7. Daly J. G., Kew A. K., Moore A. R., Olivier G. 1996; The cell surface of Aeromonas salmonicida determines in vitro survival in cultured brook trout (Salvelinus fontinalis) peritoneal macrophages. Microb Pathog 21:447–461 [CrossRef]
     [Google Scholar]
  8. Davis C. P., Avots-Avotins A. E., Fader R. C. 1981; Evidence for a bladder cell glycolipid receptor for Escherichia coli and the effect of neuraminic acid and colominic acid on adherence. Infect Immun 34:944–948
     [Google Scholar]
  9. Devrets D. A., Campbell P. A. 1991; Macrophage phagocytosis: use of fluorescence microscopy to distinguish between extracellular and intracellular bacteria. J Immunol Methods 142:31–38 [CrossRef]
     [Google Scholar]
  10. Finlay B. B., Falkow S. 1988; A comparison of microbial invasion strategies of Salmonella, Shigella and Yersinia species. In Bacteria–Host Cell Interaction pp. 227–243Edited by Horowitz M. A. New York: Alan R. Liss;
     [Google Scholar]
  11. Galán J. E. 1994; Interactions of bacteria with non-phagocytic cells. Curr Opin Immun 6:590–595 [CrossRef]
     [Google Scholar]
  12. Garcı́a-Peñarrubia P., Cabrera L., Alvarez R., Galvez J. 1992; Effector–target interactions: saturability, affinity and binding isotherms. A study of such interactions in the human NK cell–K562 tumour cell system. J Immunol Methods 155:133–147 [CrossRef]
     [Google Scholar]
  13. Guerina N. G., Kessler T. W., Guerina V. J., Neutra M. R., Clegg H. W., Langermann S., Scannapieco F. A., Goldmann D. A. 1983; The role of pili and capsule in the pathogenesis of neonatal infection with Escherichia coli K1. J Infect Dis 148:395–405 [CrossRef]
     [Google Scholar]
  14. Gutenberger S. K., Duimstra J. R., Rohovec J. S., Fryer J. L. 1997; Intracellular survival of Renibacterium salmoninarum in trout mononuclear phagocytes. Dis Aquat Org 28:93–106 [CrossRef]
     [Google Scholar]
  15. Hawke J. P., Plakas S. M., Minton R. V., McPhearson R. M., Snider T. G., Guarino A. M. 1987; Fish pasteurellosis of cultured striped bass (Morone saxatilis) in coastal Alabama. Aquaculture 65:193–204 [CrossRef]
     [Google Scholar]
  16. Hayat M. A. 1986 Basic Techniques for Transmission Electron Microscopy Orlando: Academic Press;
     [Google Scholar]
  17. Heinzen R. A., Hayes S. F., Peacock M. G., Hackstadt T. 1993; Directional actin polymerization associated with spotted fever group Rickettsia infection of Vero cells. Infect Immun 61:1926–1935
     [Google Scholar]
  18. Huang X.-Z., Tall B., Schwan W. R., Kopecko D. J. 1998; Physical limitations on Salmonella typhi entry into cultured human intestinal epithelial cells. Infect Immun 66:2928–2937
     [Google Scholar]
  19. Jacques M., Bélanger M., Roy G., Foiry B. 1991; Adherence of Actinobacillus pleuropneumoniae to porcine tracheal epithelial cells and frozen lung sections. Vet Microbiol 27:133–143 [CrossRef]
     [Google Scholar]
  20. Jacques M., Kobisch M., Bélanger M., Dugal F. 1993; Virulence of capsulated and noncapsulated isolates of Pasteurella multocida and their adherence to porcine respiratory tract cells and mucus. Infect Immun 61:4785–4792
     [Google Scholar]
  21. Janda J. M., Abbott S. L., Oshiro L. S. 1991; Penetration and replication of Edwardsiella spp. in Hep-2 cells. Infect Immun 59:154–161
     [Google Scholar]
  22. Konkel M. E., Mead D. J., Cieplak W. Jr 1993; Kinetic and antigenic characterization of altered protein synthesis by Campylobacter jejuni during cultivation with human epithelial cells. J Infect Dis 168:948–954 [CrossRef]
     [Google Scholar]
  23. Kubota S., Kimura M., Egusa S. 1970; Studies on bacterial tuberculoidosis of the yellowtail. I. Symptomatology and histopathology. Fish Pathol 4:111–118 [CrossRef]
     [Google Scholar]
  24. Kusuda R., Salati F. 1993; Major bacterial diseases affecting mariculture in Japan. Annu Rev Fish Dis 3:69–85 [CrossRef]
     [Google Scholar]
  25. Leung K. Y., Lim T. M., Lam T. J., Sin Y. M. 1996; Morphological changes in carp epithelial cells infected with Aeromonas hydrophila. J Fish Dis 19:167–174 [CrossRef]
     [Google Scholar]
  26. Lewis P. R., Knight D. P. 1977; Staining methods for sectioned material. In Practical Methods in Electron Microscopy pp. 1–311Edited by Glauert A. M. Amsterdam: Elsevier;
     [Google Scholar]
  27. Lindhal M., Faris A., Wadstrom T., Hjerten S. 1981; A new test based on ‘‘salting out’’ to measure relative surface hydrophobicity of bacterial cells. Biochim Biophys Acta 677:471–476 [CrossRef]
     [Google Scholar]
  28. Lipuma J. L., Gilsdorf J. R. 1987; Role of capsule in adherence of Haemophilus influenzae type b to human buccal epithelial cells. Infect Immun 55:2308–2310
     [Google Scholar]
  29. Magariños B., Romalde J. L. 1997; Immunization with bacterial antigens: Pasteurellosis. In Fish Vaccinology pp. 167–177Edited by Gudding R., Lillehaug A., Midtlying P. J., Brown F. Basel: Karger;
     [Google Scholar]
  30. Magariños B., Santos Y., Romalde J. L., Rivas C., Barja J. L., Toranzo A. E. 1992; Pathogenic activities of live cells and extracellular products of the fish pathogen Pasteurella piscicida. J Gen Microbiol 138:2491–2498 [CrossRef]
     [Google Scholar]
  31. Magariños B., Romalde J. L., Lemos M. L., Barja J. L., Toranzo A. E. 1994; Iron uptake by Pasteurella piscicida and its role in pathogenicity for fish. Appl Environ Microbiol 60:2990–2998
     [Google Scholar]
  32. Magariños B., Romalde J. L., Noya M., Barja J. L., Toranzo A. E. 1996a; Adherence and invasive capacities of the fish pathogen Pasteurella piscicida. FEMS Microbiol Lett 138:29–34 [CrossRef]
     [Google Scholar]
  33. Magariños B., Bonet R., Romalde J. L., Martı́nez M. J., Congregado F., Toranzo A. E. 1996b; Influence of the capsular layer on the virulence of Pasteurella piscicida for fish. Microb Pathog 21:289–297 [CrossRef]
     [Google Scholar]
  34. Mills S. D., Finlay B. B. 1994; Comparison of Salmonella typhi and Salmonella typhimurium invasion, intracellular growth and localization in cultured human epithelial cells. Microb Pathog 17:409–423 [CrossRef]
     [Google Scholar]
  35. Nelson J. S., Kawahara E., Kawai K., Kusuda R. 1981; Macrophage infiltration in pseudotuberculosis of yellowtail, Seriola quinqueradiata. Bull Mar Sci Fish Kochi Uni 11:17–22
     [Google Scholar]
  36. Noya M., Magariños B., Lamas J. 1995a; Interactions between peritoneal exudate cells (PECs) of gilthead seabream (Sparus aurata) and Pasteurella piscicida. A morphological study. Aquaculture 131:11–21 [CrossRef]
     [Google Scholar]
  37. Noya M., Magariños B., Toranzo A. E., Lamas J. 1995b; Sequential pathology of experimental pasteurellosis in gilthead seabream Sparus aurata. A light- and electron- microscopic study. Dis Aquat Org 21:177–186 [CrossRef]
     [Google Scholar]
  38. Roberson B. S. 1990; Bacterial agglutination. In Techniques in Fish Immunology pp. 81–86Edited by Stolen J. S., Fletcher T. C., Anderson D. P., Roberson B. S., van Muiswinkel W. B. Fair Aven, NJ: SOS Publications;
     [Google Scholar]
  39. Roberts M., Jacobs R. F., Haas J. E., Smith A. L. 1984; Adherence of Haemophilus influenzae to monkey respiratory tissue in organ cultures. J Gen Microbiol 130:1437–1447
     [Google Scholar]
  40. Romalde J. L., Magariños B. 1997; Immunization with bacterial antigens: pasteurellosis. Dev Biol Stand 90:167–177
     [Google Scholar]
  41. Rubens C. E., Smith S., Hulse M., Chi E. Y., van Belle G. 1992; Respiratory epithelial cell invasion by group B streptococci. Infect Immun 60:5157–5163
     [Google Scholar]
  42. Runnels P. L., Moon H. W. 1984; Capsule reduces adherence of enterotoxigenic Escherichia coli to isolated intestinal epithelial cells of pigs. Infect Immun 45:737–740
     [Google Scholar]
  43. St Geme J. W. III, Cutter D. 1996; Influence of pili, fibrils, and capsule on in vitro adherence by Haemophilus influenzae type b. Mol Microbiol 21:21–31 [CrossRef]
     [Google Scholar]
  44. St Geme J. W. III, Falkow S. 1991; Loss of capsule expression by Haemophilus influenzae type b results in enhanced adherence to and invasion of human cells. Infect Immun 59:1325–1333
     [Google Scholar]
  45. Sakai D. K. 1981; Heat inactivation of complements and immune hemolysis reaction in rainbow trout, masu salmon, coho salmon, goldfish and tilapia. Bull Jpn Soc Sci Fish 47:565–571 [CrossRef]
     [Google Scholar]
  46. Sansonetti P. J., Ryter A., Clerc P., Maurelli A., Mounier J. 1986; Multiplication of Shigella flexneri within Hela cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis. Infect Immun 51:461–469
     [Google Scholar]
  47. Santos Y., Bandı́n I., Nieto T. P., Bruno D. W., Ellis A. E., Toranzo A. E. 1990; Comparison of the cell surface hydrophobicity of bacterial fish pathogens by different procedures. In Pathology in Marine Science pp. 101–115Edited by Cheng T. C., Perkins F. O. New York: Academic Press;
     [Google Scholar]
  48. Skarmeta A. M., Bandı́n I., Santos Y., Toranzo A. E. 1995; In vitro killing of Pasteurella piscicida by fish macrophages. Dis Aquat Org 23:51–57 [CrossRef]
     [Google Scholar]
  49. Small P. L. C., Isberg R. R., Falkow S. 1987; Comparison of the ability of enteroinvasive Escherichia coli, Salmonella typhimurium, Yersinia pseudotuberculosis, and Yersinia enterocolitica to enter and replicate within HEp-2 cells. Infect Immun 55:1674–1679
     [Google Scholar]
  50. Tan E., Low K. W., Wong W. S. F., Leung K. Y. 1998; Internalization of Aeromonas hydrophila by fish epithelial cells can be inhibited with a tyrosine kinase inhibitor. Microbiology 144:299–307 [CrossRef]
     [Google Scholar]
  51. Thune R. L., Stanley L. A., Cooper R. K. 1993; Pathogenesis of Gram-negative bacterial infections in warmwater fish. Annu Rev Fish Dis 3:37–68 [CrossRef]
     [Google Scholar]
  52. Toranzo A. E., Barreiro S., Casal J. F., Figueras A., Magariños B., Barja J. L. 1991; Pasteurellosis in cultured gilthead seabream, Sparus aurata: first report in Spain. Aquaculture 99:1–15 [CrossRef]
     [Google Scholar]
  53. Virji M., Kayhty H., Ferguson D. J. P., Alexandrescu C., Moxon E. R. 1991; Interactions of Haemophilus influenzae with cultured human endothelial cells. Microb Pathog 10:231–245 [CrossRef]
     [Google Scholar]
  54. Wang X. H., Oon H. L., Ho G. W. P., Wong W. S. F., Lim T. M., Leung K. Y. 1998; Internalization and cytotoxicity are important virulence mechanisms in vibrio–fish epithelial cell interactions. Microbiology 144:2987–3002 [CrossRef]
     [Google Scholar]
  55. Watanabe S., Sasaki J., Wanda T., Tanaka Y., Otsuka N. 1988; Low gel temperature agarose encapsulation of small specimens for electron microscopy. J Electron Microsc 37:89–91
     [Google Scholar]
  56. Wolke R. E. 1975; Pathology of bacterial and fungal diseases affecting fish. In The Pathology of Fishes pp. 33–116Edited by Ribelin W. E., Migaki G. Madison, WI: University of Wisconsin Press;
     [Google Scholar]
  57. Yoshida T., Inglis V., Misawa N., Kruger R., Sakai M. 1997; In vitro adhesion of Pasteurella piscicida to cultured fish cells. J Fish Dis 20:77–80 [CrossRef]
     [Google Scholar]
  58. Zierler M. K., Galán J. E. 1995; Paradigms in bacterial entry into host cells. In Virulence Mechanisms of Bacterial Pathogens pp. 21–31Edited by Roth J. A., Bolin C. A., Brogden K. A., Minion F. C., Wannemuehler M. J. Washington, DC: American Society for Microbiology;
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-146-1-21
Loading
Invasion of fish epithelial cells by Photobacterium damselae subsp. piscicida: evidence for receptor specificity, and effect of capsule and serum
Microbiology 146, 21 (2000); https://doi.org/10.1099/00221287-146-1-21
/content/journal/micro/10.1099/00221287-146-1-21
/content/journal/micro/10.1099/00221287-146-1-21
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error