Linking cholinesterase inhibition with behavioural changes in the sea snail Gibbula umbilicalis: Effects of the organophosphate pesticide chlorpyrifos

Inhibition of acetylcholinesterase (AChE) activity has been widely used to assess the exposure and effects of anticholinergic environmental contaminants in several species. The aim of this study was to investigate if sublethal concentrations of the organophosphorous pesticide chlorpyrifos (CPF), a well-known AChE inhibitor, would also affect cholinesterases (ChE) in Gibbula umbilicalis and if this inhibition would result in an alteration of its behaviour, in an attempt to link the effects observed at the cellular level with effects at higher levels of ecological relevance. The biochemical properties of ChEs in this species were first characterized through the assessment of different enzymatic forms present in the sea snail, using different substrates and selective inhibitors. The results suggest that G. umbilicalis possess ChEs with characteristics of typical AChE, which should be the main form present. Additionally, in vitro and in vivo effects of CPF on AChE activity were investigated, along with effects on snails' behaviour: the ability of the snails to move/turn after exposure to the contaminant (flipping test). As expected, CPF inhibited AChE activity both in vitro and in vivo conditions. Moreover, the link between AChE activity inhibition and adverse effects on behavioural changes was established: AChE inhibition was positively correlated with the flipping test, indicating a mechanistic relationship between the two endpoints determined in in vivo exposures. This study highlights the importance of linking biochemical endpoints such as AChE activity with higher level endpoints like behavioural alterations, increasing the ecological relevance of the effects observed.