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Measurement of Crassostrea gigas hemocyte oxidative metabolism by flow cytometry and the inhibiting capacity of pathogenic vibrios

Abstract : A flow cytometric method to measure the production of oxidative metabolism products was adapted for use with Crassostrea gigas hemocytes. The method is based upon the oxidation, by hydrogen peroxide (H2O2), of intracellular 2',7'-dichlorofluorescin (DCFH) to green-fluorescent dichlorofluorescein. Activation of the respiratory burst (RB) was tested using phorbol myristate acetate with no success. By contrast, activation by zymosan particles increased oxidation of DCFH in C. gigas hemocytes, mainly granulocytes, and optimization tests showed a good response with 20 zymosan particles per hemocyte. Anti-aggregant solution, used to prevent hemocytes from clumping during bleeding, inhibited the RB activity measured by DCFH oxidation. The flow cytometric method developed during this work was used to evaluate the DCFH oxidation-inhibiting capacity of four strains of vibrio bacteria, known or suspected to be pathogenic for bivalves.
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https://hal.archives-ouvertes.fr/hal-00616961
Contributor : Christophe Lambert Connect in order to contact the contributor
Submitted on : Thursday, August 25, 2011 - 11:21:54 AM
Last modification on : Monday, December 13, 2021 - 8:30:03 AM
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Christophe Lambert, Philippe Soudant, Gwénaëlle Choquet, Christine Paillard. Measurement of Crassostrea gigas hemocyte oxidative metabolism by flow cytometry and the inhibiting capacity of pathogenic vibrios. Fish and Shellfish Immunology, Elsevier, 2003, 15, pp.225-240. ⟨hal-00616961⟩

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