Temefos and six of its metabolites were tested for their capacity to inhibit the in vitro activity of brain acetylcholinesterase (AChE) of Fundulus heteroclitus. While temefos was not inhibitory at levels up to 10.7 mM in brain homogenate samples, its metabolites were active within the range of 2 X 10(-4)mM to 1.26 mM in causing a 50% reduction in the enzyme activity. Exposure of F. heteroclitus to temefos under laboratory conditions caused a reduction in AChE activity, which was proportional to the pesticide concentration and the exposure period. Visible symptoms of organophosphate poisoning were apparent only after the AChE inhibition reached 80%. F. heteroclitus and Cyprinidon variegatus exposed to 10 biweekly applications of temefos granules in the field showed no inhibition of brain AChE. However, exposure of F. heteroclitus to biweekly applications (four) of temefos emulsion caused a reduction in the enzyme (50%), but only in the pre-third application samples. A gradual increase in brain AChE occurred both in F. heteroclitus and C. variegatus as the season progressed from April to October.