Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1993 Jun;59(6):1848–1854. doi: 10.1128/aem.59.6.1848-1854.1993

Indigenous Bacteria in Hemolymph and Tissues of Marine Bivalves at Low Temperatures

Jan A Olafsen 1,*, Helene V Mikkelsen 1, Hanne M Giæver 1, Geir Høvik Hansen 1,†
PMCID: PMC182171  PMID: 16348962

Abstract

Hemolymph and soft tissues of Pacific oysters (Crassostrea gigas) kept in sand-filtered seawater at temperatures between 1 and 8°C were normally found to contain bacteria, with viable counts (CFU) in hemolymph in the range 1.4 × 102 to 5.6 × 102 bacteria per ml. Pseudomonas, Alteromonas, Vibrio, and Aeromonas organisms dominated, with a smaller variety of morphologically different unidentified strains. Hemolymph and soft tissues of horse mussels (Modiolus modiolus), locally collected from a 6- to 10-m depth in the sea at temperatures between 4 and 6°C, also contained bacteria. The CFU in horse mussel hemolymph was of the same magnitude as that in oysters (mean, 2.6 × 104), and the bacterial flora was dominated by Pseudomonas (61.3%), Vibrio (27.0%), and Aeromonas (11.7%) organisms. In soft tissues of horse mussels, a mean CFU of 2.9 × 104 bacteria per g was found, with Vibrio (38.5%), Pseudomonas (33.0%), and Aeromonas (28.5%) constituting the major genera. After the challenge of oysters in seawater at 4°C to the psychrotrophic fish pathogen Vibrio salmonicida (strains NCIMB 2245 from Scotland and TEO 84001 from Norway) and a commensal Aeromonas sp. isolated from oysters, the viable count in hemolymph increased 1,000-fold to about 105 bacteria per ml. In soft tissues, about a 1,000-fold increase in CFU to 6 × 107 was observed. V. salmonicida NCIMB 2245 invaded hemolymph and soft tissues after 14 days and dominated these compartments after 41 days, whereas strain TEO 84001 did not invade soft tissues to the same extent. Challenge with V. salmonicida NCIMB 2245 resulted in 100% mortality, whereas about 50% of the oysters survived challenge with the Norwegian strain, TEO 84001. The commensal Aeromonas sp. invaded hemolymph and soft tissues and caused 100% mortality. Oyster hemolymph contained agglutinins for Vibrio anguillarum but not for V. salmonicida, whereas we did not find agglutinins for either of these bacteria in horse mussels. Agglutinins for horse and human erythrocytes were found in hemolymph from both animals. We found no differences in agglutinin titers in oysters from different Norwegian locations, and long-term challenge with bacteria in seawater did not result in changes of agglutinin activity. These studies demonstrate that bacteria exist in hemolymph and soft tissues of marine bivalves at temperatures below 8°C. Increased bacterial numbers in seawater at 4°C result in augmented invasion of bacteria in hemolymph and soft tissues. V. salmonicida, a bacterium pathogenic for fish at low temperatures, invades bivalve hemolymph and soft tissues, and thus bivalves may serve as a reservoir for pathogens of fish at low seawater temperatures.

Full text

PDF
1848

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baross J., Liston J. Occurrence of Vibrio parahaemolyticus and related hemolytic vibrios in marine environments of Washington State. Appl Microbiol. 1970 Aug;20(2):179–186. doi: 10.1128/am.20.2.179-186.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. COLWELL R. R., LISTON J. Microbiology of shellfish. Bacteriological study of the natural flora of Pacific oysters (Crassostrea gigas). Appl Microbiol. 1960 Mar;8:104–109. doi: 10.1128/am.8.2.104-109.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cheng T. C., Marchalonis J. J., Vasta G. R. Role of molluscan lectins in recognition processes. Prog Clin Biol Res. 1984;157:1–15. [PubMed] [Google Scholar]
  4. Colwell R. R., Sparks A. K. Properties of Pseudomonas enalia, a marine bacterium pathogenic for the invertebrate Crassostrea gigas (Thunberg). Appl Microbiol. 1967 Sep;15(5):980–986. doi: 10.1128/am.15.5.980-986.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DePaola A., Hopkins L. H., Peeler J. T., Wentz B., McPhearson R. M. Incidence of Vibrio parahaemolyticus in U.S. coastal waters and oysters. Appl Environ Microbiol. 1990 Aug;56(8):2299–2302. doi: 10.1128/aem.56.8.2299-2302.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Enger O., Husevåg B., Goksøyr J. Presence of the fish pathogen Vibrio salmonicida in fish farm sediments. Appl Environ Microbiol. 1989 Nov;55(11):2815–2818. doi: 10.1128/aem.55.11.2815-2818.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Farley C. A. Neoplasms in estuarine mollusks and approaches to ascertain causes. Ann N Y Acad Sci. 1978 Sep 29;298:225–232. doi: 10.1111/j.1749-6632.1977.tb19267.x. [DOI] [PubMed] [Google Scholar]
  8. Fisher W. S., DiNuzzo A. R. Agglutination of bacteria and erythrocytes by serum from six species of marine molluscs. J Invertebr Pathol. 1991 May;57(3):380–394. doi: 10.1016/0022-2011(91)90142-d. [DOI] [PubMed] [Google Scholar]
  9. Hansen G. H., Olafsen J. A. Bacterial Colonization of Cod (Gadus morhua L.) and Halibut (Hippoglossus hippoglossus) Eggs in Marine Aquaculture. Appl Environ Microbiol. 1989 Jun;55(6):1435–1446. doi: 10.1128/aem.55.6.1435-1446.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hoff K. A. Survival of Vibrio anguillarum and Vibrio salmonicida at different salinities. Appl Environ Microbiol. 1989 Jul;55(7):1775–1786. doi: 10.1128/aem.55.7.1775-1786.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kelly M. T., Dinuzzo A. Uptake and clearance of Vibrio vulnificus from Gulf coast oysters (Crassostrea virginica). Appl Environ Microbiol. 1985 Dec;50(6):1548–1549. doi: 10.1128/aem.50.6.1548-1549.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kelly M. T. Effect of temperature and salinity on Vibrio (Beneckea) vulnificus occurrence in a Gulf Coast environment. Appl Environ Microbiol. 1982 Oct;44(4):820–824. doi: 10.1128/aem.44.4.820-824.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Olafsen J. A., Fletcher T. C., Grant P. T. Agglutinin activity in Pacific oyster (Crassostrea gigas) hemolymph following in vivo Vibrio anguillarum challenge. Dev Comp Immunol. 1992 Mar-Jun;16(2-3):123–138. doi: 10.1016/0145-305x(92)90013-3. [DOI] [PubMed] [Google Scholar]
  14. Stein E. A., Younai S., Cooper E. L. Hemagglutinins and bacterial agglutinins of earthworms. Prog Clin Biol Res. 1987;233:79–89. [PubMed] [Google Scholar]
  15. Tamplin M. L., Capers G. M. Persistence of Vibrio vulnificus in tissues of Gulf Coast oysters, Crassostrea virginica, exposed to seawater disinfected with UV light. Appl Environ Microbiol. 1992 May;58(5):1506–1510. doi: 10.1128/aem.58.5.1506-1510.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Tamplin M. L., Fisher W. S. Occurrence and characteristics of agglutination of Vibrio cholerae by serum from the eastern oyster, Crassostrea virginica. Appl Environ Microbiol. 1989 Nov;55(11):2882–2887. doi: 10.1128/aem.55.11.2882-2887.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tamplin M., Rodrick G. E., Blake N. J., Cuba T. Isolation and characterization of Vibrio vulnificus from two Florida estuaries. Appl Environ Microbiol. 1982 Dec;44(6):1466–1470. doi: 10.1128/aem.44.6.1466-1470.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tubiash H. S., Chanley P. E., Leifson E. Bacillary necrosis, a disease of larval and juvenile bivalve mollusks. I. Etiology and epizootiology. J Bacteriol. 1965 Oct;90(4):1036–1044. doi: 10.1128/jb.90.4.1036-1044.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tubiash H. S., Colwell R. R., Sakazaki R. Marine vibrios associated with bacillary necrosis, a disease of larval and juvenile bivalve mollusks. J Bacteriol. 1970 Jul;103(1):271–272. doi: 10.1128/jb.103.1.271-272.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tubiash H. S. Soft-shell clam, Mya arenaria, a convenient laboratory animal for screening pathogens of bivalve mollusks. Appl Microbiol. 1971 Sep;22(3):321–324. doi: 10.1128/am.22.3.321-324.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES