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Ultra-sensitive conductometric detection of pesticides based on inhibition of esterase activity in Arthrospira platensis
2013
Tekaya, Nadèje | Saiapina, Olga | Ben Ouada, Hatem | Lagarde, Florence | Ben Ouada, Hafedh | Jaffrezic-Renault, N. (Nicole)
Enzymatic conductometric biosensor, using immobilized Arthrospira platensis cells on gold interdigitated electrodes, for the detection of pesticides in water, was elaborated. Cholinesterase activity (AChE) was inhibited by pesticides and a variation of the local conductivity was measured after addition of the substrate acetylthiocholine chloride (AChCl). The Michaelis–Menten constant (Km) was evaluated to be 1.8 mM through a calibration curve of AChCl. Inhibition of AChE was observed with paraoxon-methyl, parathion-methyl, triazine and diuron with a detection limit of 10−18 M, 10−20 M, 10−20 M and 10−12 M, respectively and the half maximal inhibitory concentration (IC50) was determined at 10−16 M, 10−20 M, 10−18 M and 10−06 M, respectively. An important decrease of response time τ90% was recorded for AChE response towards AChCl after 30 min cell exposure to pesticides. Scanning electron microscopy images revealed a degradation of the cell surface in presence of pesticides at 10−06 M.
Show more [+] Less [-]Comparative toxicity of chlorpyrifos, diazinon, malathion and their oxon derivatives to larval Rana boylii
2007
Sparling, D.W. | Fellers, G.
Organophosphorus pesticides (OPs) are ubiquitous in the environment and are highly toxic to amphibians. They deactivate cholinesterase, resulting in neurological dysfunction. Most chemicals in this group require oxidative desulfuration to achieve their greatest cholinesterase-inhibiting potencies. Oxon derivatives are formed within liver cells but also by bacterial decay of parental pesticides. This study examines the toxicity of chlorpyrifos, malathion and diazinon and their oxons on the foothill yellow-legged frog (Rana boylii). R. boylii is exposed to agricultural pesticides in the California Central Valley. Median lethal concentrations of the parental forms during a 96 h exposure were 3.00 mg/L (24 h) for chlorpyrifos, 2.14 mg/L for malathion and 7.49 mg/L for diazinon. Corresponding oxons were 10 to 100 times more toxic than their parental forms. We conclude that environmental concentrations of these pesticides can be harmful to R. boylii populations. Laboratory tests on the toxicity of OP insecticides and their oxons suggest that they may be acutely lethal to amphibians at ecologically relevant concentrations.
Show more [+] Less [-]Synergistic effects caused by atrazine and terbuthylazine on chlorpyrifos toxicity to early-life stages of the zebrafish Danio rerio
2013
Pérez, Joanne | Domingues, Inês | Monteiro, Marta | Soares, Amadeu M. V. M. | Loureiro, Susana
This study examined the effects of three widely used pesticides that have been previously detected in aquatic systems neighbouring agricultural fields on the early-life stages of the zebrafish Danio rerio. Tests involving single exposures and binary combinations of the s-triazine herbicides (atrazine and terbuthylazine) and the organophosphate insecticide chlorpyrifos were performed. Several endpoints, such as swimming behaviour, morphological abnormalities and mortality, were studied. In addition, the inhibition of acetylcholinesterase (AChE) activity was investigated in order to evaluate the mode of action and toxicity of chlorpyrifos in the presence of these herbicides. Results indicate that both binary mixtures elicited synergistic responses on the swimming behaviour of zebrafish larvae. Moreover, although the herbicides were not effective inhibitors of the AChE on their own, a synergistic inhibition of the enzyme activity was obtained by exposure to mixtures with chlorpyrifos. We observed a correlation between impairment of swimming behaviour of the larvae and inhibition of AChE activity. This study supports previous studies concerning the risk assessment of mixtures since the toxicity may be underestimated when looking only at the single toxicants and not their mixtures.
Show more [+] Less [-]Alteration of cholinesterase activity as possible mechanism of silver nanoparticle toxicity
2014
Šinko, Goran | Vinković Vrček, Ivana | Goessler, Walter | Leitinger, Gerd | Dijanošić, Adriana | Miljanić, Snežana
Due to their broad-spectrum antimicrobial activity, silver nanoparticles (AgNPs) have been used in a large number of commercial and medical products. Such proliferated AgNP production poses toxicological and environmental issues which need to be addressed. The present study aimed to investigate the effects of AgNPs on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), important enzymes in areas of neurobiology, toxicology and pharmacology. Three different AgNPs, prepared by the chemical reduction using trisodium citrate, hydroxylamine hydrochloride (Cl-AgNPs), and borohydride following stabilization with poly(vinyl alcohol), were purified and characterised with respect to their sizes, shapes and optical properties. Their inhibition potential on AChE and BChE was evaluated in vitro using an enzyme assay with o-nitrophenyl acetate or o-nitrophenyl butyrate as substrates, respectively. All three studied AgNPs were reversible inhibitors of ChEs. Among tested nanoparticles, Cl-AgNP was found to be the most potent inhibitor of both AChE and BChE. Although the detailed mechanism by which the AgNPs inhibit esterase activities remains unknown, structural perturbation of the enzyme may be the common mode of ChE inhibition by AgNPs.
Show more [+] Less [-]Differential expression profile of membrane proteins in Aplysia pleural–pedal ganglia under the stress of methyl parathion
2014
Chen, Ying-Ying | Huang, Lin | Zhang, Yong | Ke, Cai-Huan | Huang, He-Qing
This study was aimed to analyze the alteration of membrane protein profiles in Aplysia juliana Quoy & Gaimard (A. juliana) pleural–pedal ganglia under MP exposure. Both the results of GC–MS analysis and the activity assay of acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CAT) reveal that MP toxicological effects on Aplysia left and right pleural–pedal ganglia are different under 7 and 14 days of exposure. Therefore, Aplysia were subjected for exposure at two concentrations (1 and 2 mg/l) of MP for 7 and 14 days for membrane proteomic study. As a result, 19 and 14 protein spots were differentially expressed in A. juliana left pleural–pedal ganglia under 7 and 14 days treatment, and 20 and 14 protein spots found with differential expressions in their right ganglia under the same treatment, respectively. Several proteins with expression variations were detected from both the left and right pleural–pedal ganglia; however, most proteins have distinctive expressions, indicating different mechanisms might be involved in initiating MP toxicology in left and right ganglia. Among the total differential protein spots obtained, 29 proteins were classed as membrane proteins. These proteins are mainly involved in the metabolism process, cell redox homeostasis, signal transduction, immunology, intracellular transport and catalysis, indicating MP toxicity in mollusks seems to be complex and diverse. Some differentially expressed proteins were further confirmed by Western blotting and quantitative real-time PCR. These results might provide renovated insights to reveal the mechanism of MP-induced neurotoxicity, and the novel candidate biomarkers might have potential application for environmental evaluation of MP pollution level.
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