Cholinesterase characterization and effects of the environmental contaminants chlorpyrifos and carbofuran on two species
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RESEARCH ARTICLE
Cholinesterase characterization and effects of the environmental contaminants chlorpyrifos and carbofuran on two species of marine crabs, Carcinus maenas and Pachygrapsus marmoratus Ana Filipa Nogueira 1 & Bruno Nunes 1 Received: 25 March 2020 / Accepted: 30 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Among the most frequent targets for toxic effects of modern pesticides, namely organophosphates and carbamates, one may find cholinesterases (ChEs). ChEs exist in a wide variety of animals and have been used actively to discriminate among the environmental effects of different pollutant groups, including the aforementioned pesticides. This study had three purposes, namely (i) identifying the ChE forms present in tissues (eyes and walking legs muscle) of two crab species, Carcinus maenas and Pachygrapsus marmoratus; to (ii) determine the in vitro toxicological effects, and (iii) compare the sensitivity of such enzymatic forms towards commonly used anti-ChE pesticides, namely the organophosphate chlorpyrifos and the carbamate carbofuran. Our results showed that there was not a clear preference for any of the tested substrates in any of the tissues from both species. Furthermore, the ChE activity was almost completely suppressed following incubation with eserine and with the specific inhibitor BW284C51 in all tissues from both species. In vitro exposure to chlorpyrifos promoted a significant decrease in ChE activity in both species. Furthermore, the ChE activity was completely suppressed following incubation with carbofuran and chlorpyrifos. These results suggest that the major ChE forms present in tissues of both crab species show intermediate structural properties and activity patterns, halfway between classic acetylcholinesterase and pseudocholinesterases. However, the sensitivity of the found forms towards ChE inhibitors was established, and the responsiveness of such forms towards common anti-ChE chemicals was established. Both tested species seem to be promising test organisms to be used in marine and coastal scenarios of putative contaminations by anti-ChE chemicals, considering the here reported patterns of response. Keywords Cholinesterases (ChEs) . Pesticides . Chlorpyrifos . Carbofuran . Biomarker . In vitro assays
Introduction ChEs (ChEs) are classified as esterases and have the capacity of hydrolyzing carboxylic esters. ChEs show a preference for the hydrolysis of choline esters, therefore they can be differentiated from other esterases (Nunes et al. 2005; Rodrigues et al. 2011). ChEs hydrolytic activity can be significantly diminished by exposure to specific chemical agents, such as pesticides, namely those from the organophosphate and
Responsible Editor: Cinta Porte * Bruno Nunes [email protected] 1
Centro de Estudos do Ambiente e do Mar, CESAM, Universidade de Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal
carbamate classes. These pesticides form a covalent bond with the ChE’s active site, resulting in a stable enzyme-substrate
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