Validation of a microtitre plate ELISA for measuring cortisol in fish and comparison of stress responses of rainbow trout (Oncorhynchus mykiss) and lake trout (Salvelinus namaycush)

摘要: A microtitre plate enzyme-linked immunoassay (ELISA) was validated for measuring serum cortisol in rainbow trout (Oncorhynchus mykiss) and lake (Salvelinus namaycush). The ELISA uses a highly specific rabbit antibody raised against cortisol-3-carboxymethyloxime (CMO)/bovine albumin (BSA), conjugate of horseradish peroxidase (HRP) also linked to the 3-position by CMO bridge. chromatogenic substrate is 3,3′,5,5′-tetramethylbenzidine (TMB). standard curve linear (logit/log) from lower limit sensitivity assay (2 pg/well = approx. 100 pg/ml serum) approximately 1000 pg/well. satisfied strictest criteria specificity, reproducibility (interassay coefficient variation 98%). Changes cortisol, glucose chloride were measured following an acute 1-min handling stressor. In both species, levels rose rapidly peaks 300 ng/ml 1 h post-stress, slowly returned baseline over next 48 h. Glucose faster than but reached similar peak 135 mg/dl at 6 post-stress species. Chloride declined between 3 although showed greater more prolonged hypochloremia trout. Within week administration stressors, 19 ( ∼ 17% fish experiment) 2 2%) died. Most which died had been sampled when their low, suggesting that survival rates stocked may be increased if they are rehandled before or after this period hypochloremia.