Physiological and biochemical responses of Mediterranean green crab, Carcinus aestuarii, to different environmental stressors: Evaluation of hemocyte toxicity and its possible effects on immune response

Abstract

Effects of natural stressors such as copper (Cu2+), temperature, hypoxia, chloroform and adrenaline on physiological and biochemical responses were investigated in the Mediterranean green crab Carcinus aestuarii from tidal shallow waters of Narta Lagoon, Albania. For this purpose, hemolymph glucose levels, total and differential hemocyte count, in normal and eye-stalked individuals, exposed to above mentioned stressors like, were assessed. In addition, lysosomal membrane stability was evaluated as biomarker of hemocyte toxicity, with possible implications on crab immune response. Hemolymph glucose levels were significantly increased in all treatment groups with 1.25-to 3.5-fold above baseline levels of 37.8 ± 2.7 mgdL−1. Response times were being manifested within 30–120 min following exposure and recovery happened within 2 h of restoration of pretreatment conditions. Total hemocyte count (THC) and differential hemocyte count (DCH) showed a significant decrease for all stressors, except for copper, were an increase of semi-granular hemocyte fraction were recorded. Meanwhile, significant reduction of neutral red retention time (NRRT), in both eyestalk-ablated and exposed animals, were recorded, indicated the loss of hemocyte lysosomal membrane integrity. The responsiveness of hemolymph blood levels to all stressors, the decrease in total hemocyte count, as well as the loss of lysosomal membrane integrity demonstrated that exposure to environmentally realistic stressors placed a heavy metabolic load on C. aestuarii, modulating their immune competence and overall physiological wellness. Overall, results suggest that monitoring cellular and biochemical parameters like hemolymph glucose titres, TCH, DHC and NRRT, may be useful and sensitive means of evaluating the crustacean's ability to cope with the wide variety of environmental stressors through modulation of the immune parameters.