Immunotoxicological effects of environmentally relevant concentrations (10, 23, 50, 100 μg/l) of atrazine were studied in Lymnaea stagnalis. Individual hemolymph sampling was performed at 0, 24, 48, 72, 96, 168, 336, 504 and 672 h during exposure. Every atrazine concentration induced a significant increase in the mean number of circulating hemocytes, without any concentration–response relation. A peak (1.6-fold increase) of hemocyte density was observed after 96 h of exposure. After 504 h, the number of hemocytes remained higher only in the snails exposed to the two highest concentrations. Granulocytes contributed most to the increase in hemocyte density in herbicide-exposed snails. Both short- (24 and 96 h) and long-term (504 h) exposures resulted in significant inhibition of hemocyte phagocytic activity upon E. coli. Over the long-term, phagocytosis recovered for the two lowest concentrations. After 504 h of exposure, every herbicide level resulted in a significant reduction of reactive oxygen species production in E. coli-stimulated hemocytes, which was not observed for short-term exposures.

An investigation was made to find analytical method for anilazine and propazine remaining in agricultural products. Since anilazine was found to be decomposed enzymatically when applied on cabbage, radish, etc., phosphoric acid was added to the sample to suppress the enzymatic activity before subjecting the sample to an extraction with acetone. The extract was re-extracted into n-hexane, cleaned up with a Florisil column, and finally subjected to a determination with ECD-GC. Propazine was found to be insusceptable to any enzymatic decomposition, but it was subjected to the same acetone extraction and n-hexane re-extraction as with anilazine, separated from anilazine with a Florisil column, and subjected to a determination with NPD-GC. The detection limits of these methods are 0.01 micro g/g for both anilazine and propazine. The addition-recovery experiments conducted resulted in recoveries of over 79 and 82% with coefficient of variation of less than 5.6 and 5.5% for alilazine and propazine, respectively (n=5). The present methods were applied to actual agricultural products, with neither anilazine nor propazine detected (n=22).