Adverse effects of chlorpyrifos and cypermethrin mixture on physiological alterations and cholinesterase expression on Nile tilapia (Oreochromis niloticus)
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Abstract
The purpose of this study was to study the effects of chlorpyrifos mixed with cypermethrin insecticides on physiological changes and the application of cholinesterase (ChE) expression as a biomarker of exposure in Nile tilapia. Based on the results, it was found that the substances affected the physiology and behavior of exposed fish. Mortality rates increased with an increasing in concentration and exposure time. Enzyme activity in the fish brain decreased with an increasing in exposure time, which was significantly different from other organs (P < 0.05). From Western blot analysis, we found acetylcholinesterase (AChE), having a molecular weight of 71 kDa, which could be detected in the brain, muscle and gill. In contrast, butyrylcholinesterase (BuChE) could be detected in plasma at 85 kDa. The appropriate concentrations of ChE, causing cross–reactivity, were 12, 10 and 10 μg/μL in the brain, muscle and gill, respectively. In plasma, the appropriate concentration of protein applied was 3 μg/μL. The detection limits for ChE detection of dot blot technique were consecutively at 5, 2.5, 0.31 and 1.25 μg/μL in brain, muscle, plasma and gill. Based on this study, ChE could be used as a biomarker of exposure for chlorpyrifos mixed with cypermethrin insecticides by using antibody techniques, i.e., Western blot and dot blot techniques for detection.
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