Glyphosate-based herbicide (GBH) causes damage in embryo-larval stages of zebrafish (Danio rerio).

Glyphosate-Based Herbicides (GBH) show risks to the environment and also to aquatic organisms, such as fish. The present work aimed to evaluate the effects of GBH and Pure Glyphosate (PG) exposure on Danio rerio embryos at drinking water concentrations. Zebrafish embryos were exposed to 250, 500, and 1000 µg L-1 of Roundup Original DI® and pure glyphosate for 96 h. Glyphosate concentration in water, parameters physicochemical water, survival, hatching rate, heart rate, malformations, behavior, and biomarkers were evaluated. We verified that at 6 h post-fertilization (hpf), animals exposed to GBH 500 showed decreased survival as compared to the control. The hatching rate increased in all groups exposed to GBH at 48 hpf as compared to the control group. The embryos exposed did not present changes in the spontaneous movement and touch response. Exposed groups to GBH demonstrated a higher number of malformations in fish embryos as compared to the control. Most malformations were: pericardial edema, yolk sac edema, body malformations, and curvature of the spine. In heart rate, bradycardia occurred in groups exposed, as predicted due to cardiac abnormalities. As biochemical endpoints, we observed a decrease in Glutathione S-transferase (GBH 250, GBH 500 and PG 250) and Acetylcholinesterase (GBH 250 and PG 250) activity. No differences were found between the groups in the concentration of protein, Total Antioxidant Capacity Against Peroxyl Radicals, Lipid peroxidation, Reactive Oxygen Species, Non-protein thiols, and Catalase. In conclusion, the damage in all evaluated stages of development was aggravated by survival and malformations. Therefore, the large-scale use of GBHs, coupled with the permissiveness of its presence could be the cause damage to the aquatic environment affecting the embryonic development of non-target organisms.

Can Vitamin C Supplementation Improve the Antioxidant Capacity of Rhamdia quelen Fish Exposed to Atrazine?

Atrazine (ATZ) is one of the pesticides mostly widely used in Brazil; several studies have shown the toxic effects of this herbicide on aquatic organisms such as fish. Thus, it is absolutely necessary finding alternatives to protect the health of fish, mainly of species commercially important for aquaculture, which may be exposed to atrazine deriving from agricultural runoff. The aim of the current study was to investigate interactions between dietary supplementation with vitamin C (Vit C) antioxidant and exposure to ATZ in Rhamdia quelen fish exposed to this herbicide. R. quelen specimens were divided into four groups: (1) CTRL, (2) VitC, (3) ATZ, (4) ATZ + VitC. Groups 3 and 4 were exposed to ATZ (10 µg L-1) for 96 h, after 30 days of VitC supplementation (1 g kg-1). Liver samples were collected for biomarker assays. Group 4 was the only group presenting decreased protein carbonyl content. Non-protein thiol (NPSH) levels were significantly higher in groups VitC, ATZ and ATZ + VitC than in CTRL. Group ATZ + VitC presented significant increase in glutatione-peroxidase (GPx) activity in comparison to the other investigated groups. Ascorbic acid (AA) levels were significantly higher in group VitC and lower in group ATZ. Therefore, interactions between herbicide ATZ and dietary supplementation with Vit C have shown biochemical changes in R. quelen fish. Thus, dietary supplements with adequate amounts of Vit C can be added as antioxidants to the diet of fish bred in aquaculture systems in order to protect them from exposure to ATZ.

Biochemical and Behavioral Responses in Zebrafish Exposed to Imidacloprid Oxidative Damage and Antioxidant Responses.

Imidacloprid (IMI) is an insecticide used worldwide, a neonicotinoid that could cause toxicity in non-target organisms. Zebrafish (Danio rerio) is a model organism widely used in different fields of research such as behavioral studies, biochemical parameters as well as neurotoxicity research. Here, we investigate whether the exposure to three concentrations (0.15, 15, and 45 µg/L) of IMI for 96 h alters responses in zebrafish. Oxidative stress parameters and acetylcholinesterase activity (AChE) as well as the behavioral responses of locomotion were measured. IMI exposure decreased distance traveled in fish exposed to the 45 µg/L. In the exploratory activity, time spent and transitions to the top area of the water column decreased in fish exposed to all concentrations of IMI. In addition, exposures to 45 and 15 µg/L of IMI decreased episodes of erratic movement in the zebrafish. Exposures to IMI at a concentration of 45 µg/L decreased the time spent in erratic movements and increased the time spent with no movement (i.e., "freezing"). Glutathione S-transferase (GST) activity was increased in the brain of zebrafish exposed for 96 h to concentrations of 0.15 and 45 µg/L. Brain AChE activity was reduced and the levels of carbonyl protein (CP) increased in brain of zebrafish at concentrations of 15 and 45 µg/L. Lipid peroxidation measured by TBARS and, also non-protein thiols (NPSH) did not show any variation in the brain of zebrafish exposed to IMI. Changes in the activity of cholinergic neurotransmitters in the brain tissues of zebrafish indicate IMI toxicity. Exposures of fish over 96 h to IMI at a nominal concentration of 45 µg/L caused more extensive sublethal responses in zebrafish, but this concentration is well above those expected in the aquatic environment. Studies are warranted to evaluate the effects on behavior and biomarker responses in fish exposed over longer periods to IMI at environmentally relevant concentrations.

Ecological impacts of pesticides on Astyanax jacuhiensis (Characiformes: Characidae) from the Uruguay river, Brazil.

Brazilian freshwater ecosystems are continuously exposed to pesticides and domestic sewage. The Uruguay River was chosen for this study because of its international importance, as it flows through Brazil, Argentina, and Uruguay. It receives contaminants such as pesticides and domestic residues. Thus, the aim of this study to assess the accumulation of pesticides in muscle of the fish Astyanax jacuhiensis, its biochemical responses, and the presence of pesticides in water. In total, seven pesticides were registered in water from both river sites. Eight pesticides were detected in fish muscle. The biochemical responses showed that brain lipid peroxidation (LPO) and protein carbonyl (PC) in A. jacuhiensis were higher in the summer. Muscle showed the highest LPO levels in the spring and the highest PC in the summer. Liver LPO and PC levels were higher in the spring and summer. In the gills, the PC was higher in the spring and the LPO in the spring and winter. In the brain and in the gills, glutathione-S-transferase activity was high in the summer and autumn. Catalase activity was lower during the winter and spring. Non-protein thiol (NPSH) levels were lower in the brain in the winter and spring. Muscle tissue showed lower NPSH in the winter (site 1). Liver NPSH showed increased levels in liver in the spring and winter (site 2). The biochemical results clearly is related to pesticides and/or to the presence of other contaminants in the water such as metals or domestic sewage. The accumulation of pesticides in fish muscle added evidence that pesticides have been used in the area surrounding the Uruguay River. In conclusion, the biomarkers assayed in the present study could be used in future investigations considering other sampling sites along Uruguay River.

Raising the water temperature: consequences in behavior and biochemical biomarkers of the freshwater crab Aegla longirostri (Crustacea, Anomura).

Understanding how temperature alone affects biomarkers commonly used in ecotoxicology studies and biomonitoring programs is important to obtain a more real response in field studies, especially in freshwater. Thus, we analyzed the behavioral responses, the lethality, and the biochemical biomarkers in the freshwater crustacean Aegla longirostri at different water temperatures. Animals were exposed under laboratory conditions, to 18 °C, 21 °C, 24 °C, and 26 °C for 48 h. There were significant changes in biochemical parameters in different tissues (hepatopancreas, gills, and muscle) and in the behavioral tests in A. longirostri. Hepatopancreas was especially affected by the elevation of temperature, as showed by the high levels of carbonyl proteins. The activity of acetylcholinesterase increased in a temperature-dependent manner in muscle. Glutathione S-transferase activity decreased with the elevation of temperature in all tissues sampled. The results obtained in this study indicate that when assessing the health of polluted limnic ecosystems through the use of organisms in situ, the intrinsic effect of abiotic factors, such as temperature, on biomarkers must be considered.

Organic and conventional agriculture: Conventional rice farming causes biochemical changes in Astyanax lacustris.

World food production is directly related to human population growth. Chemicals are constantly applied to pest control in crops to increase productivity. Therefore, sustainable alternatives are needed to reduce environmental impacts. The biochemical responses in liver and muscle of Astyanax lacustris collected in different rice planting systems were analysed. Ten fish were collected in organic rice cultivation systems and conventional as well as water and sediment for pesticide analysis. In water from conventional system, bentazon (56.1 µg L-1), fipronil (0.226 µg L-1) and propoxur (0.141 µg L-1) were found, while azoxystrobin and quinclorac were below the limit of quantification (LOQ). There were no pesticides in the sediment from the conventional system. In the water of the organic system, only propoxur (below the LOQ) was registered. Metalaxil (0.025 µg kg-1) were verified in the sediment, while diphenoconazole, dinoxifene and tebuconazole were below the LOQ. The presence of these pesticides in crops with an organic production system may be related to proximity to crops to conventional production systems. Besides this, the amount is very low as compared with conventional system. Muscle glycogen, protein and amino acid levels were higher in fish collected in organic ponds. Lactate and ammonia levels were higher in conventional cultures. Lipids and proteins had greater oxidative damage in both tissues in the conventional system. Although pesticides were detected in organic sediments, the parameters of metabolic and oxidative damage were probably related by exposure to higher concentrations of pesticide in the water or by the lower oxygen content of conventional systems. However, fish mortality was not observed during collections. Despite being an anthropic environment, areas of organic cultivation seem to present better conditions for the survival of A. lacustris.

Biochemical Responses in Freshwater Fish Exposed to Insecticide Propoxur.

Although designed to control pests selectively, there is some evidence that environmental contamination by pesticides increases risks for humans and wildlife. In the present study, we evaluated biomarkers of oxidative stress in Astyanax jacuhiensis exposed to (5, 15 and 30 µg L-1) of carbamate Propoxur (PPX) for 96 h. Glutathione S-transferase (GST) in liver and gills showed reduced activity in all PPX concentrations tested. Acetylcholinesterase (AChE) activities reduced in brain and muscle at concentrations 15 and 30 µg L-1 of PPX. Lipid peroxidation (LPO) and hydrogen peroxide (HP) had no significant differences. In the brain, protein carbonyl (PC) increased in all groups treated with PPX. Although PPX is a selective pesticide, it causes oxidative damage and enzyme alteration in fish. This study pointed out some biomarkers that could be used to assess effects of environmentally relevant concentrations of pesticides, and infer about studies using fish as bioindicator.

Seasonal implications on toxicity biomarkers of Loricariichthys anus (Valenciennes, 1835) from a subtropical reservoir.

Cropping systems based on intensive land use and continuous application of agricultural chemicals inflict a threat to aquatic organisms since these substances will inevitably be carried in to water bodies where they can accumulate, particularly in lentic sites. Pesticides exposure in aquatic animals can cause changes that can be quantified through biomarkers. Thus, this study aimed to investigate the effects of season on oxidative stress and neurotoxicity biomarkers in Loricariichthys anus from a subtropical reservoir surrounded by agricultural areas in southern Brazil. Ten armored catfish were collected from six sites in February and August 2016. Pesticides present in the water, sediment and muscle were identified and quantified. No pesticides were detected either in sediment nor in muscle. During the winter, the water contained atrazine, imidacloprid, simazine, azoxystrobin, and propoxur; however, in summer, only atrazine was present in the water. In the winter, there was an increase in the hepatic GST activity and in GPx that kept lipid peroxidation (TBARS) constant and, in the summer, there was an increase in metallothioneins levels. In the gills, variables related to summer were possibly responsible for the elevation of GST, GPx and TBARS; during the winter, there was greater carbonylation of proteins. In the winter, the increased AChE activity in brain and muscle tissue was related to carbonylation of proteins in brain. Although the amount of pesticides detected in the water was low, chronic exposure in addition to environmental variations can cause direct and indirect effects on L. anus population.