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http://dx.doi.org/10.4217/OPR.2012.34.2.137

Assessment of Immune Parameters of the Wild Pacific Oyster (Crassostrea gigas) using a Flow Cytometry and Neutral Red Retention Assay  

Hong, Hyun-Ki (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute, Jeju National University)
Kang, Hyun-Sil (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute, Jeju National University)
Kim, Young-Ok (South Sea Research Institute, KIOST)
Choi, Kwang-Sik (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute, Jeju National University)
Publication Information
Ocean and Polar Research / v.34, no.2, 2012 , pp. 137-149 More about this Journal
Abstract
Hemocyte parameters of the wild Pacific oyster Crassostrea gigas inhabiting intertidal zones in small bays (Gwangyang and Jinhae Bay) on the southern coast of Korea were evaluated using flow cytometry and neutral red retention (NRR) assay. Morphological features, cell count, mortality, DNA damage, phagocytosis, and lysosomal membrane stability of hemocytes were analyzed. Three types of hemocytes were identified in the oyster hemolymph: granulocytes, hyalinocytes, and blast-like cells. Immune related functions of hemocyte including phagocytosis and lysosomal membrane stability were significantly different among the study areas (P<0.05), while cell count, mortality, and DNA damage of hemocytes were not significantly different. In Gwangyang Bay, phagocytosis of granulocytes and lysosomal membrane stability of oyster hemocytes inhabiting inside bay were significantly lower than those of oyster hemocytes in outside bay (P<0.05), indicating that oysters in inside bay of Gwangyang were relatively suppressed the immunological function in hemocytes. Contrary to Gwangyang Bay, immune parameters of oyster hemocytes in Jinhae Bay not showed the difference between sampling sites. In conclusion, flow cytometry and NRR assay using oyster hemocyte has a powerful tool to investigate the cell level in a short time due to no-preprocessing of material.
Keywords
Crassostrea gigas; flow cytometry; neutral red retention assay; hemocyte; DNA damage; phagocytosis; lysosomal membrane stability;
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