Abstract To address the potential role of cholinesterase enzymes in the invasive capacity of species, the present study investigated ChE activity in the invasive freshwater bivalve Anodonta woodiana (Lea, 1834) comparing it with that of the indigenous species, Anodonta sp. (Linnaeus, 1758). The invasive capacity of pests has often been linked to their ecological plasticity and high intrinsic genetic variability; however the role played by molecular and cellular mechanisms, generally known as an organism's response to pollution, is unclear. Different substrates and selective ChE enzyme inhibitors were investigated in digestive gland, foot, gonad, adductor muscle and gill tissues while sensitivity to four organophosphate (OP) insecticides was investigated in vitro only in adductor muscle. The invasive species ( A. woodiana) showed significantly greater (at least one order of magnitude) ChE activity than the endemic species ( Anodonta sp.) ( p < 0.05) using acetylthiocholine (ASCh) as substrate and the activity was more widely distributed in tissues involved in movement (adductor muscle and foot), respiration, feeding (gills) and reproduction (gonads). Moreover, only the invasive species, A. woodiana, showed detectable ChE (vs. ASCh) activity in gill tissue. No substrate specificity was observed in any tissue of either species as already described for other bivalve species. ChE activity was not inhibited by Iso-OMPA but showed high sensitivity to BW248c51 and eserine. Both species showed moderate to low sensitivities in vitro to OP insecticides in the range 10 − 7 –10 − 2 M. Calculated IC 50 for fenitrothion and chlorpyrifos was in the range 10 − 6 –10 − 3 M in muscle of A. woodiana while a higher inhibition was observed for fenitrothion (10 − 7 M) and lower for chlorpyrifos (10 − 2 M) in the indigenous species Anodonta sp. Similar IC 50 of 10 − 5 –10 − 6 M were observed for DFP and azamethiphos in both species. The hypotheses of other authors that acetylcholinesterase (AChE) is involved in the control of many essential functions, such as frontal ciliary activity of gill epithelium, temperature resistance, ciliary activity for transport of suspended particulate, valve opening and embryo development, suggest that the high catalytic efficiency of the invasive species may endow it with a competitive advantage over the endemic species. In view of the peculiar reproductive strategy of these mussels, higher ChE vs. ASCh activity in gonads of the invasive species could also favour glochidium production and embryo development under a wider range of environmental conditions.