Acetylcholinesterase activity in Gammarus fossarum (Crustacea Amphipoda) Intrinsic variability, reference levels, and a reliable tool for field surveys.

The appropriate use of an enzyme activity as a biomarker requires good knowledge of its basal level and its natural variability related to intrinsic biotic and environmental abiotic factors. In view of using whole-body acetylcholinesterase (AChE) activity in Gammarus fossarum as a reliable biomarker of exposure to anti-cholinesterase agents in aquatic ecosystems, (i) the effects of the main biotic (sex, reproductive status, and weight) and abiotic (water temperature) factors on the basal activity level of this enzyme were measured in the laboratory and (ii) the spatio-temporal variability of basal enzyme activity was followed in wild populations over a 1-year period. The results show no direct effect of sex. However, significant differences in AChE activity were observed between females depending on gonadal and embryonic development. A strong negative correlation between the AChE activity levels and organism body weight was observed. Indeed, AChE activity decreases drastically during the early life stages and tends to stabilise in larger individuals. These reports led us to select a standard organism (male; weight range, 15-20mg) to minimise inter-individual variability. No effect of temperature on basal AChE activity was observed in the laboratory for the tested range (6-24 degrees C). Similarly, no spatio-temporal change relative to season or the physico-chemical characteristics of the water (such as conductivity and temperature) was recorded during the field survey. On the basis of field-collected data, we defined the standard organism having a reference activity level with minimal and maximal threshold values. Finally, the value of AChE activity normalisation by protein contents is discussed.