Morphological and functional characterization of clam Ruditapes philippinarum haemocytes

Clam Ruditapes philippinarum is one of the most important commercial aquaculture species in China. The haemocytes play vital roles in internal defense of the calm. In this investigation, classification and immune functions of R. philippinarum haemocytes were identified. The haemocyte density was (8.28 1.42) x 10(6)/mL and two major haemocyte types basophilic hyalinocytes and eosinophilic granulocytes were recognized based on the presence or absence of granules and staining affinities of their cytoplasm. Granulocytes were the most common cell type (73.08 +/- 3.23%). The hyalinocytes and granulocytes could be divided into eight subtypes respectively according to N/C ratio as well as the nucleus shape and number by light microscope. Fourteen types of granules were identified and the multivesicular body and R-body were first found in bivalve, moreover, transmission electron microscope observation was consistent with the results from light microscope. Also eight different external forms of haemocytes could be identified by scanning electron microscope. Both granulocytes and hyalinocytes showed the abilities of phagocytosis and reactive oxygen species (ROS) production which were higher in granulocytes than that in hyalinocytes. The phagocytic rate of the total haemocytes and the granulocytes was about 45.06% and 40.74% respectively. The ROS production of the total haemocytes and granulocytes was 58.7% and 51.19% respectively. Although the hyalinocytes showed less ability in phagocytosis and ROS production, they played important roles in agglutination. This investigation provided a fundamental knowledge for future study of the immune function of haemocytes in clam R. philippinarum.