Dimethoate affects cholinesterases in Folsomia candida and their locomotion - false negative results of an avoidance behaviour test

The main mode of action of organophosphate insecticides is to inhibit acetylcholinesterase (AChE), which causes neuromuscular paralysis leading ultimately to death. The collembolan Folsomia candida is an important and standard test species in ecotoxicology, where effects on avoidance behaviour are assessed. Being related to insects they represent potential targets of insecticides such as the organophosphate dimethoate. In the present study we exposed F. candida to dimethoate having 2 main aims: 1) to assess the ability of F. candida to avoid it, and 2) to assess its effect on the cholinergic synapses to explore the link. For the latter, several sub-steps were needed: a) to characterise the existing ChE types and b) assess ChE activity (via exposure in vitro and in vivo). No avoidance was observed within the tested concentration range (0-0.32-1-3.2-10-32mg/kg), in fact an apparent "attraction" (more animals on the spiked side) was observed. As expected, there was a significant decrease of AChE activities (AChE being the main ChE type) with an increase of dimethoate dose (IC50=1.4mg/kg). Further, post-exposure video records showed that organisms were still alive in the spiked soil but lacked the locomotion ability (immobilised). The AChE inhibition correlated positively with immobilisation. Hence, this observation also showed that the apparent "attraction" behaviour observed in the avoidance test is rather a direct effect of not being able to escape due to paralysis hence a false-negative avoidance. This can constitute a confounding factor in an avoidance behaviour test and consequent interpretation, which is not accounted for at present.This study was supported by a funding from FEDER through COMPETE e Programa Operacional Factores de Competitividade and by the national funding through FCT-Fundação para a Ciência e Tecnologia, within the research project FUBIA FCOMP-01-0124-FEDER-008651 (Ref. PTDC/AAC-CLI/103719/2008).publishe