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Microplastics (MPs) are currently one of the main problems of marine pollution, being found in all environmental matrices. Due to their size, they can be ingested by organisms directly (from the environment) or indirectly (with their prey). The objective of this study was to analyze the occurrence, abundance, concentration, and chemical nature of MPs present in the gastrointestinal tract of two fish species, Patagonotothen guntheri and Patagonotothen ramsayi, both of which are key in the food web of the Marine Protected Area Namuncurá/ Banco Burdwood (MPA N/BB). The analyzed species presented high values of MPs per individual (MPs/ind.) and occurrence compared to other studies. P. guntheri tended to have a lower number of MPs/ind. and occurrence than P. ramsayi (P. guntheri: 2.50 ± 1.93 MPs/ind., 82.50  %; P. ramsayi: 3.93 ± 2.91 MPs/ind., 90.60  %). While fibers were the predominant MPs in both species, P. ramsayi had a greater number of fragments and a greater variety of MPs chemical composition than P. guntheri. The prevailing chemical composition was cellulosic material (cellulose and cellulose mixed with polyamide and polyester). Synthetic fibers and fragments such as polyester (PET), alkyd resin, polyurethane, polyethylene, polyacrylic fiber and poly(ethylene-co-vinyl acetate-co-vinyl chloride) were also found. Although both species have a generalist diet, the differences found may be due to the fact that P. guntheri has benthopelagic feeding habits while P. ramsayi has demersal-benthic. Our study is the first report on the presence and characterization of MPs in organisms relevant to food webs in the Southwest Atlantic., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Microplastics (MPs) have been identified in diverse marine invertebrates; however, there are few studies on limpets. This study aimed to evaluate and compare the presence of MPs in Nacella magellanica among sites of the Beagle Channel with different degree of anthropogenic influence. Intertidal limpets were sampled in three sites: Ushuaia Bay (UB) (inside Ushuaia city), a site with high anthropogenic influence owing to population growth, and fisheries, industrial and port activities; Ensenada Zaratiegui (EZ) and Playa Larga (PL) (west and east of Ushuaia city, respectively), both sites with a certain degree of environmental protection and low anthropogenic influence. MPs were isolated from the whole soft tissue of individuals by oxidative digestions with H 2 O 2 (30%; 1:10 w/v). MPs occurrence was 100% in organisms from UB and EZ, whereas 90% in PL. Limpets from UB showed 10 ± 6.69 MPs/ind and 2.22 ± 0.78 MPs/g ww, while in PL and EZ there were 2.90 ± 2.02 MPs/ind and 0.96 ± 0.94 MPs/g ww; and 5.60 ± 2.59 MPs/ind and 2.41 ± 1.47 MPs/g ww, respectively. Fibres were the most abundant MPs type, followed by fragments and films. Fibres and films were identified as semi-synthetic cellulose and, although the polymer matrix of fragments was masked by copper phthalocyanine, one of them could be identified as polystyrene. As expected, the highest abundance of MPs was found in UB. However, a lower abundance of particles was detected in PL (downstream of UB) than in EZ (upstream of UB). This could be due to the influence of both the main current of the Beagle Channel and fluvial discharges, reducing the buoyancy of MPs. More studies are needed to understand the contribution of regional and local features to the accumulation of MPs in these environments and their interaction with the biota., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Neurotoxic insecticides are ubiquitous in aquatic ecosystems, frequently as part of complex mixtures. Freshwater gastropods are generally underrepresented in neurotoxicity evaluations and cumulative toxicity testing. This study investigates the behavioural and biochemical effects of acute exposures to the carbamate carbaryl, the organophosphate chlorpyrifos, and the neonicotinoid acetamiprid on the freshwater gastropod Chilina gibbosa. First, we evaluated behavioural neurotoxicity and cholinesterase (ChE), carboxylesterase (CE), and glutathione S-transferase (GST) activities in acute (48h) single-chemical exposures to increasing concentrations of carbaryl (0.5-500 μg L -1 ), chlorpyrifos (10-7500 μg L -1 ), and acetamiprid (1-10000 μg L -1 ). We then studied the effects of acute (48h) exposures to binary mixtures of carbaryl and chlorpyrifos equivalent to 0.5, 1, and 1.5 ChE 48h-IC 50 . None of the insecticides caused severe behavioural neurotoxicity, except for a significant lack of adherence by 5000 μg L -1 chlorpyrifos. Carbaryl caused concentration-dependent inhibition of ChEs (NOEC 5 μg L -1 ; 48h-IC 50 45 μg L -1 ) and CEs with p-nitrophenyl butyrate as substrate (NOEC 5 μg L -1 ; 48h-IC 50 37 μg L -1 ). Chlorpyrifos caused concentration-dependent inhibition of ChEs (NOEC 50 μg L -1 ; 48h-IC 50 946 μg L -1 ) but did not affect CEs (NOEC ≥7500 μg L -1 ). Carbaryl-chlorpyrifos mixtures inhibited ChEs additively, inhibited CEs with p-nitrophenyl butyrate, and did not affect behaviour. GST activity was not affected by single or mixture exposures. Acute exposure to acetamiprid did not affect any of the endpoints evaluated. This study provides new information on carbaryl, chlorpyrifos, and acetamiprid toxicity on C. gibbosa, relevant to improve gastropod representation in ecotoxicological risk assessment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2020. Published by Elsevier Inc.)

This work aimed to study in the freshwater gastropod Planorbarius corneus the effects of acute (2 days) and subchronic (14 days) exposures to an environmental concentration of the organophosphate azinphos-methyl on different reproductive parameters, offspring survival and B-esterase activities in gonads and in the whole organism soft tissue. The acute exposure inhibited only carboxylesterase activity in both tissues while the subchronic exposure also inhibited cholinesterase activity, decreased the number of hatched-eggs and increased offspring lethality (92%). On the other hand, B-esterases in gonads were more effective biomarkers than B-esterases in the whole organism due their inhibition appeared earlier in time (cholinesterase activity) and their activity remained inhibited for a longer time (carboxylesterase activity) when recovery studies were performed. We concluded that B-esterases and reproductive parameters can be used as effect biomarkers of aquatic contamination with azinphos-methyl. Our studies showed that a 14 days exposure to an environmental concentration of azinphos-methyl produced severe signs of toxicity in adult organisms, egg masses and juveniles that could cause negative effects at the population level in contaminated environments., (Copyright © 2018 Elsevier Inc. 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., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The use of a battery of biomarkers, especially those more closely related to species integrity, is desired for more complete ecotoxicological assessments of the effects of pesticide contamination on aquatic organisms. The phosphorodithioate azinphos-methyl has been intensively used in agriculture worldwide and have been found in the habitat of Chilina gibbosa, a freshwater snail endemic to South America. This snail has been proposed as a good model organism for ecotoxicity bioassays on the basis of studies focused mainly on enzymatic responses in whole tissue homogenates. Our aim was to evaluate the effect of an acute 48 h exposure to an environmental concentration of azinphos-methyl on C. gibbosa hemolymph enzymatic activity and cellular immune response. Our results show that cholinesterase activity was strongly inhibited (94%) in hemolymph of exposed snails. Carboxylesterase activity measured with p-nitrophenyl butyrate and glutathione S-transferase activity were augmented 47% and 89% respectively after exposure. No differences were found for hemolymph carboxylesterase activity measured with p-nitrophenyl acetate. These results differ from those reported for whole tissue homogenates and reveal that tissue-specific responses of enzymatic biomarkers exist in this species. Regarding immune cell response, hemocytes were identified for the first time for C. gibbosa. Their viability and phagocytic activity decreased after azinphos-methyl exposure although total number of circulating cells did not differ between treatments. We conclude that concentrations of azinphos-methyl that can be found in the environment can compromise both hemolymph cholinesterase activity and the immune system of C. gibbosa. Furthermore, we propose that carboxylesterase and glutathione S-transferase activities measured in hemolymph and hemocyte viability and phagocytic activity could be incorporated as sensitive biomarkers to evaluate the effects of pesticide exposure on this and related species., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Azinphos-methyl belongs to the class of organophosphate insecticides which are recognized for their anticholinesterase action. It is one of the most frequently used insecticides in the Upper Valley of Río Negro and Río Neuquén in Argentina, where agriculture represents the second most important economic activity. It has been detected in water from this North Patagonian region throughout the year and the maximum concentration found was 22.48 μg L(-1) during the application period. Chilina gibbosa is a freshwater gastropod widely distributed in South America, particularly in Patagonia, Argentina and in Southern Chile. Toxicological studies performed with C. gibbosa in our laboratory have reported neurotoxicity signs and cholinesterase inhibition after exposure to azinphos-methyl for 48 h. Recovery studies together with characterization of the enzyme and sensitivity of the enzyme to pesticides can improve the toxicological evaluation. However, little is known about recovery patterns in organisms exposed to organophosphates. The aim of the present work was to evaluate the recovery capacity (during 21 days in pesticide-free water) of cholinesterase activity and neurotoxicity in C. gibbosa after 48 h of exposure to azinphos-methyl. Also, lethality and carboxylesterase activity were registered during the recovery period. Regarding enzyme activities, after a 48-h exposure to 20 μg L(-1) of azinphos-methyl, cholinesterases showed an inhibition of 85% with respect to control, while carboxylesterases were not affected. After 21 days in pesticide-free water, cholinesterases continued to be inhibited (70%). Severe neurotoxicity signs were observed after exposure: 82% of the snails presented lack of adherence to vessels, 11% showed weak adherence, and 96% exhibited an abnormal protrusion of the head-foot region from shell. After 21 days in pesticide-free water, only 15% of the snails presented severe signs of neurotoxicity. However, during the recovery period significant lethality (30%) was registered in treated snails. C. gibbosa is a very sensitive organism to azinphos-methyl. These snails play an important role in the structure and function of aquatic food webs in this region. Thus, a decline of this species' population would probably have an impact on aquatic and non-aquatic communities. Our results show that C. gibbosa is a relevant sentinel species for studying exposure and effects of azinphos-methyl using behavioral and biochemical biomarkers. Neurotoxic behavioral signs are very sensitive, non-destructive biomarkers, which can be easily detected for about one week after acute exposure. Cholinesterse activity is a very useful biomarker showing a high sensitivity and a slow recovery capacity increasing the possibility to indirectly detect organophosphates for long periods after a contaminant event., (Copyright © 2015 Elsevier B.V. All rights reserved.)