The protective effect of Chlorella vulgaris against diclofenac toxicity in Clarias gariepinus: haemato-immunological parameters and spleen histological features as outcome markers

Abstract

Introduction: Diclofenac (DCF) is a commonly utilized medication in the non-steroidal anti-inflammatory drug category that is released into aquatic systems in significant amounts. Chlorella vulgaris (C. vulgaris) is rich in active phytochemicals known for their haemato-immunological boosting properties. Methods: Our objective was to investigate the haemato-immunological protective properties of Chlorella in mitigating the toxic effects of DCF. Five groups of Clarias gariepinus, each comprising 36 fish, were assigned over a two-week period. The groups were assigned as follows: control group, which received a basal diet only; DCF1 group, which received a basal diet and was exposed to 20 μg/L of DCF; DCF2 group, which received a basal diet and was exposed to 10 mg/L of DCF; and Chlorella +DCF1 and Chlorella+DCF2 groups, which were exposed to the same DCF doses as Groups 2 and 3, respectively, while also being fed a diet containing 25% Chlorella. Results: Exposure to both doses of DCF significantly decreased erythrocyte count, hemoglobin content, white blood cell count, phagocytic index, and lysozyme activity, while increased eosinophil and neutrophil % in an equipotent manner. The low dose caused a more pronounced reduction in packed cell volume (PCV)% and large lymphocyte% compared to the high dose. A significant decline in platelet count was observed only with the low DCF dose, while the high dose led to a decrease in monocyte%. DCF intoxication led to a dose-related decrease in small lymphocyte% and an increase in erythrocyte morphological alterations and interleukin (IL)-6 levels. The DCF2 group exhibited a higher increase in apoptotic RBCs than the DCF1 group. Intervention with Chlorella alongside the two DCF doses significantly normalized RBC count and eosinophil %, increased PCV% and small lymphocyte%, and decreased erythrocyte abnormalities to an equal extent. Large lymphocyte% in the Chlorella+DCF1 group was successfully restored to normal levels. Phagocytic index and lysozyme activity in the supplemented groups were lower, while IL-6 levels were higher than in the DCF groups. The percentage of apoptotic cells decreased with Chlorella administration, with the Chlorella+DCF1 group showing fewer apoptotic cells than the Chlorella+DCF2 group. Histopathological deterioration and excessive collagen deposition were observed in the spleen of DCF groups, while notable improvements were seen following C. vulgaris supplementation. Conclusion: These findings suggest that dietary inclusion of C. vulgaris may antagonize the haemato-cytological abnormalities induced by DCF intoxication.