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In the Upper Valley of Río Negro and Río Neuquén in Argentina, agriculture represents the second most important economic activity. Azinphos-methyl has been found in water from this region throughout the year at a maximum concentration of 22.48 μg L(-1) during the application period. Toxicological studies on local non-target species have been performed mostly on vertebrates, while mollusks, which could be more sensitive, have not been studied so far. This work aims to characterize cholinesterase (ChE) and carboxilesterase (CE) activities of Chilina gibbosa, a freshwater gastropod native to southern Argentina and Chile. These enzymes, together with neurotoxicity signals, are evaluated herein after as sensitive biomarkers of exposure to azinphos-methyl at environmentally relevant concentrations. Effects of azinphos-methyl on antioxidant defenses: glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) and glutathione S-transferase (GST) are also studied in order to complete a set of biomarkers with different sensitivity and specificity, to propose C. gibbosa as a sentinel species. The highest specific activity was obtained with acetylthiocholine as substrate, followed by propionylthiocholine (83% in comparison to acetylthiocholine) and butyrylthiocholine (19%).The lowest Km and the highest efficiency for ChE were obtained with acetylthiocholine. Regarding CEs activities, a higher efficiency was obtained with p-nitrophenyl butyrate than with p-nitrophenyl acetate. Eserine produced significant inhibition of ChE activity (81% with 0.001 mM and 98% with 1mM) while iso-OMPA did not produce any significant effect on ChE. Our results show that C. gibbosa ChE is very sensitive to azinphos-methyl (CI50 0.02 μg L(-1)) while CEs are inhibited at higher concentrations (CI50 1,000 μg L(-1)). CEs have been reported to be more sensitive to OPs than ChEs in most of the aquatic invertebrates protecting the organisms from neurotoxic effects. In contrast, C. gibbosa, has ChE which are much more sensitive to azinphos-methyl than CEs and shows marked signs of neurotoxicity. Regarding antioxidant defenses, GSH levels were significantly increased by 0.02 and 20 μg L(-1) azinphos-methyl (80 and 103%, respectively), CAT activity was increased 85% only at 0.02 μg L(-1) and SOD and GST did not show any significant response. Since ChE activity, neurotoxicity signs, GSH and CAT are sensitive biomarkers of acute exposure to azinphos-methyl at environmental concentrations C. gibbosa could be included as sentinel species in monitoring programs of pesticide hazard in regions of Argentina and Chile., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Organophosphate insecticides (OPs) are commonly used in Argentina and around the world for pest control in food crops. They exert their toxicity through the inhibition of the enzyme acetylcholinesterase. In the present study, we aimed to evaluate biochemical and reproductive effects in Biomphalaria straminea , a freshwater gastropod naturally distributed in Argentina, of subchronic exposures to environmental azinphos-methyl concentrations (20 and 200 µg L −1 ). For biochemical parameters, adult organisms were exposed for 14 days and the activity of cholinesterases (ChEs), carboxylesterases (CEs), glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), the production of reactive oxygen species (ROS), the total antioxidant capacity (TAC), glycogen and proteins were determined. For reproductive parameters, the egg masses of B. straminea were exposed to azinphos-methyl for one month, and the hatching time and success as well as the offspring survival were registered. We found different toxic effects elicited by the insecticide on the studied biomarkers. CEs activity was significantly inhibited while CAT and GST activities, ROS production and TAC were significantly increased, with respect to the solvent control group. ChE and SOD activities and protein and glycogen contents were not altered by azinphos-methyl. The hatching time and success were not statistically different from control. Nevertheless, the offspring survival was severely affected by the insecticide. Our results show that the primary target of the insecticide (ChE) was not inhibited but CEs, GST, CAT, ROS, TAC and offspring survival were sensitive biomarkers and valuable endpoints for subchronic toxicity assessments in this species. [ABSTRACT FROM AUTHOR]

Neonicotinoids emerged as an environmentally safe alternative to previous generations of insecticides becoming one of the most widely applied in modern agriculture. Nevertheless, they have been reported to affect several non-target organisms. Most toxicity studies focus on the effects on pollinators or terrestrial invertebrates and evaluate either the active ingredient or the commercial formulation. In the present study, we aimed to assess the long-term effects of the active ingredient acetamiprid and a broadly used commercial formulation (Assail® 70) on the non-target freshwater gastropod Biomphalaria straminea using a battery of biomarkers. A 14 day-exposure of adult organisms to both active ingredient and commercial formulation increased carboxylesterase activity and glutathione content, inhibited superoxide dismutase activity and decreased reactive oxygen species levels. The commercial formulation additionally increased glutathione S-transferase activity and inhibited catalase activity. The results indicate a greater toxicity of the commercial formulation than that of the active ingredient alone. Cholinesterase activity, development and offspring survival of B. straminea were not impaired. We conclude that the toxicity of acetamiprid on this gastropod species is mainly related to effects on detoxification and oxidative metabolism responses. This study provides novel information about the adverse effects of the active ingredient and a commercial formulation of a widely used neonicotinoid on a non-target aquatic species. Image 1 • The effects of acetamiprid were evaluated on a native freshwater gastropod. • Both active ingredient and commercial formulation affected biochemical biomarkers. • The commercial formulation was more toxic than the active ingredient alone. • The insecticide did not impair development and offspring survival. [ABSTRACT FROM AUTHOR]