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Chlorpyrifos, an organophosphate insecticide widely used to control agricultural pests, poses a significant environmental threat due to its toxicity and persistence in soil and water. Our work aimed to evaluate the acute (survival) and chronic (regeneration, locomotion, and reproduction) toxicity of chlorpyrifos to the non-target freshwater planarian Girardia tigrina. The 48 h lethal concentration (LC50) of the commercial formulation, containing 480 g L-1 of chlorpyrifos, the active ingredient, was determined to be 622.8 µg a.i. L-1 for planarians. Sublethal effects were translated into a significant reduction in locomotion and delayed head regeneration (lowest observed effect concentration-LOEC = 3.88 µg a.i. L-1). Additionally, chlorpyrifos exposure did not affect planarian fecundity or fertility. Overall, this study demonstrates the potential of chlorpyrifos-based insecticides to harm natural populations of freshwater planarians at environmentally relevant concentrations. The observed toxicity emphasizes the need for stricter regulations and careful management of chlorpyrifos usage to mitigate its deleterious effects on aquatic ecosystems. By understanding the specific impacts on non-target organisms like G. tigrina, we can make more informed suggestions regarding the usage and regulation of organophosphate insecticides, ultimately promoting sustainable agricultural practices and environmental conservation.

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Chlorpyrifos (CPF) is one of the most widely used insecticides worldwide despite the fact that many authors have warned about its effects in non-target biota. The effects of CPF on anurans are well known, but the process of recovery from these effects after exposure is less explored. The aim of this study was to evaluate the duration of sublethal effects induced by environmental concentrations of CPF on Ceratophrys ornata tadpoles after exposure. The experimental design consisted of an exposure phase (96 h) in which tadpoles were individually exposed to three concentrations of CPF (0, 0.01 and 0.02 mg CPF/L) and a post-exposure phase (72 h) in which exposed tadpoles were transferred to CPF-free media. Individuals that survived the exposure phase to CPF showed neither long-term lethal effects nor long-term swimming alterations and altered prey consumption after being transferred to CPF-free media. No morphological abnormalities were observed either. However, at the end of both phases, tadpoles emitted shorter sounds with a higher dominant frequency than the tadpoles in the control group, i.e., the tadpoles did not recover their normal sounds. Thus, for the first time in this species, we have shown that effects on sounds should be prioritized as biomarkers of exposure, as they not only provide longer detection times after cessation of exposure, but also involve non-destructive methods. The following order of priority could be established for the selection of biomarkers that diagnose the health status of individuals and precede irreversible responses such as mortality: alterations in sounds > swimming alterations > prey consumption.

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The aim of this study was to evaluate the toxic effects of chlorpyrifos (CPF) at environmental concentrations on the shrimp Palaemonetes argentinus, a South American native species. Organisms were exposed to environmentally relevant concentrations of CPF (from 3.5 to 94.5ngCPFL(-1)) at laboratory conditions for 96h. A wide battery of biochemical responses including bioaccumulation, damage and defense biomarkers were measured in cephalothorax and abdomen of shrimp. The concentration of CPF was below the detection limit of the method in both body sectors (8ngCPFg(-1)ww), probably indicating fast biotransformation of the parental compound. Our results showed that CPF exposure inhibits acetylcholinesterase activity from 3.5ngCPFL(-1), a concentration below the suggested Argentinean guidelines for the protection of aquatic biota. Moreover, oxidative stress was evidenced by increased H2O2 content and increased levels of TBARs and carbonyl groups in proteins. The induction of antioxidant enzymes like catalase, glutathione S-transferase and glutathione peroxidase seems not be sufficient to prevent oxidative damages. In addition, the mobilization of α-tocopherol from abdomen to cephalothorax was observed and reported for the first time in non-reproductive condition. Likewise, a strong diminution of metallothioneins occurred in cephalothorax from the lowest CPF concentration while induction occurred from the same treatment in abdomen as an oxidative stress response. Finally, significant correlation between Integrated Biomarker Response values and exposure concentrations suggest the usefulness of P. argentinus as bioindicator of CPF exposure at concentrations as low as environmental ones.

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The immobilization of enzymes onto transducer support is a mature technology and has been successfully implemented to improve biocatalytic processes for diverse applications. However, there exists still need to design more sophisticated and specialized strategies to enhance the functional properties of the biosensors. In this work, a biosensor platform based on innovative fabrication strategy was designed, and employed for the detection of organophosphate (OP) in natural waters. The biosensor was prepared by incorporating acetylcholinesterase enzyme (AChE) to the graphite paste modified with tetracyanoquinodimethane (TCNQ) mediator, along with the use of a macroalgae (Cladaphropsis membranous) as a functional immobilization support. The novel immobilization design resulted in a synergic effect, and led to enhanced stability and sensitivity of the biosensor. The designed biosensor was used to analyze methyl parathion OP insecticide in water samples collected from a demonstrably contaminated lake of São Luis Island, Maranhão, Northeast of Brazil. Water analysis revealed that the aquatic ecosystem was polluted by sub-ppm concentrations of the OP insecticide, and a good correlation was found between values obtained through biosensor and GC-MS techniques. Our results demonstrated that macroalgae-biosensor could be used as a low-cost and sensitive screening method to detect target analyte.

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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.

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Un estudio descriptivo sobre las condiciones de salud y su relación con la exposición a plaguicidas, de los “fumigadores” de la campaña contra el dengue en el estado Aragua, fue realizado en una muestra no probabilística de 50 trabajadores, a los cuales se les aplicó una encuesta para investigar: características personales y de trabajo, conocimientos sobre insecticidas y enfermedades asociadas, además de inspeccionar locales, equipos y condiciones de trabajo. De la muestra general, 28% resultaron fumadores. Los grados de instrucción predominantes fueron primaria y secundaria (44%). El promedio de años como fumigador fue de 14,3. Sólo 40% tiene conocimiento de enfermedades producidas por organofosforados, 62% presentó antecedentes de intoxicación. 30% refirieron presentar enfermedades cardiovasculares, gastrointestinales y neurológicas. A mayor cantidad de años de labor, mayor la intensidad de signos y síntomas llegando a producir enfermedades. 38% de los niveles de Acetilcolinesterasa medidos, resultaron alterados. Las variables: grupo de edad, grado de instrucción, fumar, años de labor, falta de conocimiento de enfermedades y antecedentes de intoxicación, se asociaron a valores anormales de colinesterasa, sin presentar significancia estadística (P>0,05). La selección de los trabajadores, su capacitación, dotación de equipos y materiales adecuados, su vigilancia clínica epidemiológica, su evaluación permanente y su respectivo tratamiento deben ser revisados y tenidos en cuenta por la Corporación de Salud del estado Aragua (CORPOSALUD), haciéndolas, normas de obligatorio cumplimiento

A descriptive study on the conditions of health and their relation to the exhibition to pesticides, and of the “fumigators” of the campaign against dengue in the Aragua state, sampled 50 workers, to whom a survey was applied to investigate: personal characteristics of work, knowledge on insecticides and diseases, besides an inspection the premises, equipment and work environment. From this general sample 28% were smokers. The predominant educational level was elementary school and high school (44%). The fumigator average of working years was of 14.3. Only 40% had knowledge of diseases produced by organophosphate, 62% presented poisoning antecedents. 30% claimed to display cardiovascular, gastrointestinal and neurological diseases. The greater the number of work years, the greater the intensity of signs and symptoms of associated diseases. 38% of the measured levels of acetylcholinesterase were altered. The variables: smoking, years of work, educational level, age group and the lack of knowledge of diseases and intoxication antecedents were associated with abnormal values of acetylcholinesterase, without showing statistical significance. The selection of the workers, their qualification and suitable equipment and materials, their clinical epidemiologic monitoring, their permanent evaluation and respective treatment must be reviewed and taken into account by CORPOSALUD (Corporación de Salud del estado Aragua), making them norms of obligatory fulfillment

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