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Comparison of hemocyte parameters in the pericardial cavity and the adductor muscle sinus in the Pacific oyster, Crassostrea gigas using two types of flow cytometers

Published online by Cambridge University Press:  09 February 2008

Béatrice Gagnaire ,
Matthieu Duchemin ,
Michel Auffret ,
Hélène Thomas-Guyon  and
Tristan Renault
Béatrice Gagnaire
Affiliation:
Centre de Recherche sur les Ecosystèmes Littoraux Anthropisés, UMR 6217, avenue Michel Crépeau, 17042 La Rochelle, France IFREMER La Tremblade, Laboratoire de Génétique et Pathologie, 17390 La Tremblade, France
Matthieu Duchemin
Affiliation:
Laboratoire des Sciences de l'Environnement Marin, UMR CNRS 6539, Institut Universitaire Européen de la Mer, Université de Bretagne Occidentale, place Nicolas Copernic, 29280 Plouzané, France Institut Armand-Frappier, Institut National de la Recherche Scientifique, Campus de Pointe-Claire, 245 Hymus, Pointe-Claire, H9R 1G6, Québec, Canada
Michel Auffret
Affiliation:
Laboratoire des Sciences de l'Environnement Marin, UMR CNRS 6539, Institut Universitaire Européen de la Mer, Université de Bretagne Occidentale, place Nicolas Copernic, 29280 Plouzané, France
Hélène Thomas-Guyon
Affiliation:
Centre de Recherche sur les Ecosystèmes Littoraux Anthropisés, UMR 6217, avenue Michel Crépeau, 17042 La Rochelle, France
Tristan Renault
Affiliation:
IFREMER La Tremblade, Laboratoire de Génétique et Pathologie, 17390 La Tremblade, France
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Abstract

Parameters of hemocyte populations have been considered as relevant indicators of bivalve health and are currently used in immunotoxicological studies. Hemocytes in hemolymph can be collected by puncturing either the pericardial cavity or the adductor muscle sinus with a syringe. Flow cytometry is a methodological approach that is increasingly being used in laboratories for the study of hemocyte parameters in aquatic invertebrates. However, various protocols for hemocyte processing in laboratories equipped with different types of cytometers have been published. In this context, two flow cytometers (EPICS XL4®, Beckman Coulter and FacsCalibur®, Becton Dickinson) and two sites of hemocyte collection (pericardial cavity and adductor muscle sinus) were compared for the analysis of hemocyte parameters in the Pacific oyster, Crassostrea gigas. Hemolymph cells were analyzed in terms of their number and organelle contents. Cell mortality, phagocytosis, non specific esterase, extension of the lysosomal compartment and production of reactive oxygen species were quantified. The results showed that the phagocytic index was higher for hemocytes obtained in the muscle sinus hemolymph. The results are discussed with respect to the potential use of flow cytometry as a tool for hemocyte studies in bivalves.

Keywords

Flow cytometryHemocytesPhagocytosisCrassostrea gigas
Type
Brief Report
Information
Aquatic Living Resources , Volume 21 , Issue 1 , January 2008 , pp. 39 - 43
Copyright
© EDP Sciences, IFREMER, IRD, 2008

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References

Allam, B., Ashton-Alcox, K.A., Ford, S.E., 2001, Haemocyte parameters associated with resistance to brown ring disease in Ruditapes spp. clams. Dev. Comp. Immunol. 25, 365-375. CrossRef
Auffret M., 2005, Bivalves as models for marine immunotoxicology, In: Tryphonas H., Fournier M., Blakley B.R., Smits J.E.G., Brousseau P. (Eds.) Investigative Immunotoxicology, Models And Approaches In Immunotoxicology. CRC Press, pp. 29-48.
Auffret, M., Mujdzic, N., Corporeau, C., Moraga, D., 2002, Xenobiotic-induced immunomodulation in the European flat oyster, Ostrea edulis. Mar. Environ. Res. 54, 585-589. CrossRef
Auffret M., Oubella R., 1995, Cytological and cytometric analysis of bivalve mollusc hemocytes, In: Stolen J.S., Fletcher T.C., Smith S.A., Zelikoff J.T., Kaattari S.L., Anderson R.S., Söderhäll I., Weeks-Perkins B.A. (Eds.) Techniques in Fish Immunology - Immunology and Pathology of Aquatic Invertebrates, Fair Haven, NJ, USA, SOS Publications, pp. 55-64.
Becker C., Parker J., Hechinger M., 2001, Is forward scatter monotonic on commercial flow cytometers?, International Society for Analysis, its Applications and Computation Conference, Berlin, Germany.
Cheng T.C., 1981, Bivalves, In: Ratcliffe N.A., Rowley A.F. (Eds.) Invertebrate Blood Cells I, London, Academic Press, pp. 233-299.
Cheng, T.C., Downs, J.C., 1988, Intracellular acid phosphatase and lysozyme levels in subpopulations of oyster, Crassostrea virginica, hemocytes. J. Invertebr. Pathol. 52, 163-167. CrossRef
Chu, F.L., Volety, A.K., Hale, R.C., Huang, Y., 2002, Cellular responses and disease expression in oysters (Crassostrea virginica) exposed to suspended field contaminated sediments. Mar. Environ. Res. 53, 17-35. CrossRef
Cunha, E.M., Oliveira, M.J., Ferreira, P.G., Aguas, A.P., 2004, Mercury intake by inflammatory phagocytes: in vivo cytology of mouse macrophages and neutrophils by X-ray elemental microanalysis coupled with scanning electron microscopy. Hum. Exp. Toxicol. 23, 447-53. CrossRef
Duchemin, M., Fournier, M., Auffret, M., 2007, Seasonal variations of immune parameters in diploid and triploid Pacific oysters, Crassostrea gigas (Thunberg). Aquaculture 264, 73-81. CrossRef
Fisher S.W., 1986, Structure and functions of oyster hemocytes, In: Brehélin M. (Eds.) Immunity in Invertebrates, Berlin Heidelberg, Springer-Vrelag, pp. 25-35.
Ford, S.E., Ashton-Alcox, K.A., Kanaley, S.A., 1994, Comparative cytometric and miroscopic analyses of oyster hemocytes. J. Invertebr. Pathol. 64, 114-122. CrossRef
Fournier, M., Pellerin, J., Lebeuf, M., Brousseau, P., Morin, Y., Cyr, D., 2002, Effects of exposure of Mya arenaria and Mactromeris polynyma to contaminated marine sediments on phagocytic activity of hemocytes. Aquat. Toxicol. 59, 83-92. CrossRef
Gagnaire, B., Thomas-Guyon, H., Burgeot, T., Renault, T., 2006, Pollutant effects on Pacific oyster, Crassostrea gigas (Thunberg), hemocytes: screening of 23 molecules using flow cytometry. Cell Biol. Toxicol. 22, 1-14. CrossRef
Goedken, M., De Guise, S., 2004, Flow cytometry as a tool to quantify oyster defence mechanisms. Fish Shellfish Immunol. 16, 539-552. CrossRef
Hine, P.M., 1999, The inter-relationships of bivalve haemocytes. Fish Shellfish Immunol. 9, 367-385. CrossRef
Lambert, C., Soudant, P., Degremont, L., Delaporte, M., Moal, J., Boudry, P., Jean, F., Huvet, A., Samain, J-F., 2007, Hemocyte characteristics in families of oysters, Crassostrea gigas, selected for differential survival during summer and reared in three sites. Aquaculture 270, 276-288. CrossRef
Lehmann, A.K., Sornes, S., Halstensen, A., 2000, Phagocytosis: measurement by flow cytometry. J. Immunol. Meth. 243, 229-242. CrossRef
Moore, M.N., Lowe, D.M., Fieth, P.E.M., 1978, Lysosomal responses to experimentally injected anthracene in the digestive cells of Mytilus edulis. Mar. Biol. 48, 297-302. CrossRef
Pipe, R.K., 1992, Generation of reactive oxygen metabolites by the haemocytes of the mussel Mytilus edulis. Dev. Comp. Immunol. 16, 111-122. CrossRef
Pipe, R.K., Farley, S.R., Coles, J.A., 1997, The separation and characterisation of haemocytes from the mussel Mytilus edulis. Cell Tissue Res. 289, 537-45. CrossRef
Pipe, R.K., Coles, J.A., Carissan, F.M.M., Ramanathan, K., 1999, Copper induced immunomodulation in the marine mussel, Mytilus edulis. Aquat. Toxicol. 46, 43-54. CrossRef
Prater, M.R., Goga, R.M. Jr, Blaylock, B.L., Longstreth, J., Holladay, S.D., 2002, Single-dose topical exposure to the pyrethroid insecticide, permethrin in C57BL/6N mice: effects on thymus and spleen. Food Chem. Toxicol. 40, 1863-1873. CrossRef
Sauvé, S., Brousseau, P., Pellerin, J., Morin, Y., Senecal, L., Goudreau, P., Fournier, M., 2002, Phagocytic activity of marine and freshwater bivalves: in vitro exposure of hemocytes to metals (Ag, Cd, Hg and Zn). Aquat. Toxicol. 58, 189-200. CrossRef
Zaroogian, G., Yevich, P., 1994, The nature and function of the brown cell in Crassostrea virginica. Mar. Environ. Res. 37, 355-373. CrossRef