Combination of polycyclic aromatic hydrocarbons and temperature exposure: In vitro effects on immune response of European clam (Ruditapes decussatus)

Abstract

Marine organisms are subjected to various biotic and abiotic factors such as changes of temperature and pollutants [e.g. polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and heavy metals, which may affect their defense mechanisms. In this context, the aim was to evaluate the combined effects of temperature (20 and 30 °C) and PAHs (fluorene, phenanthrene and pyrene) at two concentrations (10−5 and 10−3 mg mL−1) on the immune responses of the European clam Ruditapes decussatus were after 24 h of in vitro exposure. Total haemocyte count, cell viability, phenoloxidase, lysozyme, alkaline phosphatase, esterase, antibacterial and agglutinating activities were measured. Exposure to high temperatures resulted in lower phosphatase alkaline activity and higher haemocyte viability and antibacterial and haemagglutinating activities, compared with the values recorded for clams maintained at low temperature. Only pyrene induced a significant decrease in haemocyte lysozyme (at 20 and 30 °C) and esterase (at 30 °C) activities. The total haemocyte count was increased by phenanthrene and pyrene at 20 °C and at 30 °C, respectively. Alkaline phosphatase activity increased when haemocytes were exposed to pyrene at 30 °C but decreased in the presence of fluorene at 20 °C. Furthermore, haemocyte viability was low in the presence of pyrene and fluorene at 20 °C and 30 °C, respectively, but was unaffected by phenanthrene. Antibacterial activity was significantly increased and no-significantly affected by the presence of pyrene and fluorene at 20 °C and 30 °C, respectively. The present study demonstrates the strong effect of PAHs and high temperature on haemocyte viability and other important immune functions, including phosphatase alkaline and antibacterial activities. Furthermore, changes in the immune parameters of European clam resulting from high temperatures may modulate the effects of PAHs and vice versa.