Embryonic Development Effects of Basagran® Herbicide in Danio Rerio: A Preliminary Study.

Bentazon (Basagran®) belongs to the chemical group of benzothiadiazinones. Thus, this study aimed to estimate the influence of herbicide bentazon (3 µg.L-1, 6 µg.L-1, 12 µg.L-1, 300 µg.L-1) in Danio rerio embryos development. The study tested environmental relevant concentrations of bentazon as well as the limit established for drinking water (300 µg.L-1) in Brazil. We performed behavioral and developmental analyzes during 96 h of exposure. The bentazon measurements after experimental period showed reduction ranging from 5.0 to 18.93% between exposed groups. Our results showed significant differences in the heart rate, which was significantly higher in groups exposed to all bentazon concentrations compared to control groups. The absence of alterations in the behavioral parameters showing that the herbicide bentazon at the concentrations tested had few adverse effects on the development and behavior of the Danio rerio embryos. Considering the toxic point of view, there is a chance that bentazon acts together with other environmental contaminants as an additive or synergistic way.

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.

Effects of isolated and combined exposures of Boana curupi (Anura: Hylidae) tadpoles to environmental doses of trichlorfon and ultraviolet radiation.

The biodiversity collapse strongly affects the amphibian group and many factors have been pointed out as catalytic agents. It is estimated that several events in the amphibian population decline worldwide may have been caused by the interaction of multiple drivers. Thus, this study aimed to evaluate the stressful effects of the exposure to environmental doses of trichlorfon (TCF) pesticide (0.5 µg/L; and an additional 100-fold concentration of 50 µg/L) and ultraviolet radiation (UV) (184.0 kJ/m² of UVA and 3.4 kJ/m² of UVB, which correspond to 5% of the daily dose) in tadpoles of the Boana curupi species (Anura: Hylidae). The isolated and combined exposures to TCF happened within 24 h of acute treatments under laboratory-controlled conditions. In the combined treatments, we adopted three different moments (M) of tadpole irradiation from the beginning of the exposures to TCF (0 h - M1; 12 h - M2; and 24 h - M3). Then, we evaluated tadpole survival, change in morphological characters, induction of apoptotic cells, lipid peroxidation (LPO), protein carbonyl content (PCC), glutathione S-transferase (GST), non-protein thiols (NPSH), and acetylcholinesterase (AChE), as well as the induction of genomic DNA (gDNA) damage. UVB treatment alone resulted in high mortality, along with a high level of apoptosis induction. Both UVA, UVB, and TCF increased LPO, PC, and AChE, while decreased GST activity. Regarding co-exposures, the most striking effect was observed in the interaction between UVB and TCF, which surprisingly decreased UVB-induced tadpole mortality, apoptosis, and gDNA damage. These results reinforce the B. curupi sensitivity to solar UVB radiation and indicate a complex response in face of UVB interaction with TCF, which may be related to activation of DNA repair pathways and/or inhibition of apoptosis, decreasing UVB-induced tadpole mortality.

Effects of glyphosate on zebrafish: a systematic review and meta-analysis.

Glyphosate herbicide is widely used in worldwide crop production. Consequently, its active ingredient, surfactants, and adjuvants commonly reach the aquatic ecosystem, thereby harming the biota. An investigation into how this herbicide affects aquatic species is important, especially in fish, as they have the ability to absorb and concentrate toxins. We aimed to evaluate the effects of glyphosate on the embryonic, larval and adult stages of zebrafish (Danio rerio), an appreciable organismal model. In this sense, we performed a meta-analysis using published articles from online databases (PubMed and ScienceDirect), which covered studies published until 2022. From a massive compilation of studies evaluating the effects of active substance glyphosate and Glyphosate-Based Herbicides (GBH) on zebrafish, we selected 36 studies used in downstream analyses. Overall, we report that glyphosate affects developmental stages and demonstrates toxicity and damage in zebrafish. We observed that embryos exposed to glyphosate exhibit increased mortality. There was also an increase in the number of morphological abnormalities related to yolk sac oedema, pericardial oedema, spinal curvature and body malformations, and a decrease in body size was observed. Furthermore, there was a decrease in the number of beats. The biochemical results demonstrated an increase in reactive oxygen species and antioxidant capacity against peroxyl radicals in the gills. The literature shows that glyphosate decreased the distance covered and the mean speed of the animals and increased the number of rotations. We concluded that glyphosate causes damage in the embryonic, larval and adult stages of this species. These results are valid for zebrafish and can be applied to other freshwater fish species. Graphical abstract.

The roles of calcium and salinity in protecting against physiological symptoms of waterborne zinc toxicity in the euryhaline killifish (Fundulus heteroclitus).

In fresh water, environmental Ca ameliorates Zn toxicity because Ca2+ and Zn2+ compete for uptake at the gills. Zn toxicity is also lower in sea water, but it is unclear whether this is due to increased Ca2+ concentration, and/or to the other ions present at higher salinity. Using the euryhaline killifish, we evaluated the relative roles of Ca2+ (as CaNO3) versus the other ions contributing to salinity in protecting against physiological symptoms of Zn2+ toxicity. Killifish were exposed to a sublethal level of Zn (500 µg/L, as ZnSO4) for 96 h in either fresh water (0 % salinity) at low (1 mmol/L) and High Ca (10 mmol/L) or 35 ppt sea water (100 % salinity) at low (1 mmol/L) and High Ca (10 mmol/L). At 0 % salinity, High Ca partly or completely protected against the following effects of Zn seen at Low Ca: elevated plasma Zn, hypocalcaemia, inhibited unidirectional Ca2+ influx, inhibited branchial Na+/K+ATPase and Ca2+ATPase activities, and oxidative stress in gills, liver, intestine, and muscle. At 100 % salinity, in the presence of 1 mmol/L (Low Ca), Zn caused no disturbances in most of these same parameters, showing that the "non-Ca" components of sea water alone provided complete protection. However, for a few endpoints (inhibited intestinal Ca2+ATPase activity, oxidative stress in gill and liver), High Ca (10 mmol/L) was needed to provide full protection against Zn in 100 % salinity. There was no instance where the combination of 100 % salinity and High Ca failed to provide complete protection against Zn-induced disturbances in sea water.

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.

Acute Silver Catfish (Rhamdia quelen) Exposure to Chlorantraniliprole Insecticide.

The aim of the current study is to investigate whether silver catfish (Rhamdia quelen) individuals exposed to commercial formulation of the chlorantraniliprole insecticide used in rice crops present changes in biochemical parameters. Fifty-four (54) silver catfish individuals were distributed in six units per tank (n = 6/repetition; triplicate/treatment) and subjected to the following treatments: T1-control, without insecticide; T2 (0.02 µg/L of insecticide) and T3 (0.20 µg/L of insecticide). Exposure time lasted 24 or 96 h, and it was followed by 96 h recovery in pesticide-free water. Results have indicated biochemical changes in cortisol, glucose, lactate and plasma protein levels, as well as few ionic changes in animals' gills during the exposure and recovery periods. Chlorantraniliprole incidence in water resulted in some biochemical changes in silver catfish specimens' plasma and gills throughout the acute exposure protocol (sub-lethal dose). Thus, chlorantraniliprole insecticide has caused osmoregulatory and/or biochemical imbalance in the investigated species under the herein adopted laboratory conditions; these changes did not get back to normal levels even after specimens were left to recover for 96 h in clean water.

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.

Environmentally relevant pesticides induce biochemical changes in Nile tilapia (Oreochromis niloticus).

The high demand for food consequently increases the entry of agricultural residues into water resources, and this phenomenon can affect non-target organisms in different ways. Environmentally relevant pesticide effects (per se or in combinations) are scarce in the scientific literature. Therefore, the aim of this study was to investigate: (1) the presence of pesticide residues in an important Brazilian source of water supply and power generation (Jacuí river), during 1 year of monitoring. (2) in a laboratory study verify the effects of the most frequently, herbicide, fungicide, and insecticide found in Jacuí river (individualized or in a mixture) on biochemical parameters in different tissues of Oreochromis niloticus. Twenty pesticide residues were detected in superficial water samples, and two of them are banned in Brazilian territory. Atrazine (0.56 µg L-1), azoxystrobin (0.024 µg L-1), and imidacloprid (0.11 µg L-1) were the most frequently herbicide, fungicide, and insecticide, respectively, found in the river and were used in the laboratory assay. O. niloticus exposed to the pesticide mixture exhibited more biochemical effects than individualized exposure groups. This response can be a result of the combined pesticide effects, culminating in an additive or synergistic effect, depending on the biomarker. In individual exposure groups, atrazine presented the most pronounced alterations, followed by azoxystrobin and imidacloprid. Overall, pesticide exposure increased levels of oxidative stress parameters, reduced antioxidant enzyme activities, and induced acetylcholinesterase activity. These findings highlight the threat to aquatic organisms which may be exposed to a miscellaneous of toxic compounds in the environment.

A mixture of pesticides at environmental concentrations induces oxidative stress and cholinergic effects in the neotropical fish Rhamdia quelen.

The insecticides imidacloprid (IMI), a neonicotinoid, and propoxur (PRO), an N-methylcarbamate compound, are pesticides widely used throughout the world. Although they are not used together to combat pests, both are often found in freshwater near agricultural areas. Thereby, the goal of this study was to evaluate the additive effects of IMI and PRO mixtures at environmental concentrations in relation to isolated compounds on Rhamdia quelen, a neotropical fish. The fish was exposed to IMI (0.11 µg/L), PRO (0.039 µg/L), or Mix (0.11 µg/L IMI plus 0.039 µg/L PRO) during 96 h. Glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), acetylcholinesterase (AChE) activities were determined. To verify oxidative damage thiobarbituric acid reactive substances (TBARS), protein carbonyl (PC), reactive oxygen species contents (ROS), antioxidant capacity against peroxides (ACAP) were determined in gills, liver, brain and muscle. The results shows that a mixture of these pesticides at environmental concentrations inhibited acetylcholinesterase activity in the brain and induced oxidative damage in all analyzed tissues. These results reinforce the hypothesis that mixture of contaminants present in environment could induce additive or synergistic effects on fish species.

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.

Influence of pesticides and abiotic conditions on biochemical biomarkers in Aegla aff. longirostri (crustacea, anomura): Implications for conservation.

Freshwater ecosystems are constantly threatened by the advance of agricultural activities. Abiotic variables (such as temperature, ammonia, and nitrite) and contaminants (e.g. pesticides) can potentially interact, increasing metabolism and the absorption of toxic substances, which can alter the ability of organisms to establish adequate stress responses. This study aimed to verify which pesticides were most frequently found and in the greatest quantities in low-order streams, and whether the combination of these pesticides with the abiotic variables altered the biological metabolism of aeglids. These freshwater crustaceans are important shredders that inhabit low-order streams and are sensitive to disturbances and/or abrupt environmental variations. The animals were exposed in situ in four streams (reference site and sites 1, 2, and 3). The reference site is a preserved stream with no apparent anthropogenic interference where aeglids still occur, while the other sites no longer exhibit populations of these animals and are influenced by agricultural activities. The exposure was performed bimonthly from November 2017 to September 2018 and lasted 96 h. Measured abiotic data and water samples were collected through all days of exposure. The analyzed biochemical parameters were acetylcholinesterase activity in muscle; and glutathione S-transferase, lipid peroxidation, protein carbonylation, non-protein thiols, antioxidant capacity against peroxides, and reactive oxygen species (ROS) in muscle, gills, and hepatopancreas. We found 24 active principles of pesticides, the most frequently being clomazone, atrazine, and propoxur. Bentazone was present at the highest amounts. The parameters evaluated in this study, including biochemical biomarkers and abiotic factors measured from the water, provided a separation of the months as a function of environmental conditions. There was a difference in activity and biomarker levels throughout the year within the same site and in some months between sites. The greater concentration or variety of pesticides associated with extreme abiotic (very high temperatures) data generated increased oxidative stress, with high levels of protein damage and considerable lipid damage in all tissues, as well as elevation in ROS, even with high levels of antioxidant capacity and non-protein thiols. With these data, we intend to warn about the risks of exposure to these environmental conditions by trying to contribute to the preservation of limnic fauna, especially aeglid crabs, because most species are under some degree of threat.

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.

Seasonal factors driving biochemical biomarkers in two fish species from a subtropical reservoir in southern Brazil: An integrated approach.

Reservoirs are lentic man-made waterbodies resulting from river damming processes. Pollutants coming from adjacent areas can accumulate in the water and sediment of these modified freshwater environments. Fish are often found in reservoirs occupying several trophic niches. Biochemical biomarkers are early warning signals of environmental disturbance to an organism. It is essential to understand how pollutants, abiotic variables and biochemical biomarker responses behave throughout the seasons to implement biomonitoring programs. Loricariichthys anus and Geophagus brasiliensis were collected, and abiotic variables were seasonally measured for one year, at six sampling sites in Passo Real reservoir, in a subtropical region of Southern Brazil. Biochemical biomarkers were analyzed in four tissues of both fish species, as well as metal and pesticide concentrations in the reservoir's water and sediment. Redundancy analysis (RDA) was carried out to find the temporal relationship between biomarkers and environmental variables. RDA has clearly shown the separation of seasons for both species. Azoxystrobin, simazine and propoxur were the pesticides mostly contributing to the variation, whereas metals had lesser contribution to it. Seasonality appears to be the main factor explaining biomarkers' variability. PERMANOVA has confirmed the effect of temperature and dissolved oxygen on biomarkers of both fish species. Thus, it is hard to differentiate if the fluctuation in biomarkers' responses only reflects the normal state of organisms or it is a biological consequence from negative effects of fish exposure to several types of pollution (sewage, pesticides, and fertilizers) entering this aquatic system. In this study, to circumvent the seasonality issue on biomonitoring, the analysis of biomarkers on these fish should not be carried out in organs directly affected by temperature (such as liver and gills), or during reproduction periods (mainly in Spring).

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.

Toxicological response of silver catfish (Rhamdia quelen) after acute exposure to a commercial insecticide containing thiamethoxam.

This study assessed the hematological, enzymatic and osmoregulatory responses of silver catfish (Rhamdia quelen) exposed to sublethal concentrations (1.125 and 3.750 µg/L) of a commercial thiamethoxam-containing insecticide used on rice crops. Groups of 6 fish per tank (in triplicate, n = 3, total 54 fish) were exposed for up to 96 h to different concentrations of the compound. After this period, fish were placed in clean water for 48 h. Two fish from each tank (6 per treatment) that had been exposed to the insecticide for 24 h were anesthetized with eugenol and blood was collected to evaluate hematological and biochemical parameters. Blood, liver and muscle were collected for determination of metabolic parameters, plasma cortisol, Cl-, Na+ and K+ levels and H+-ATPase and Na+/K+-ATPase activity in the gill. H+-ATPase activity was higher in fish exposed to 1.125 µg/L insecticide at 24 h compared to control (0.0 µg/L). Differences in cortisol levels were evidenced throughout the experimental period. These results indicated that exposure to the insecticide changed the hematological, biochemical and metabolic profile of the animals, suggesting concern about environmental safety. Therefore, we discourage the use of this pesticide in areas that come into contact with water bodies inhabited by fish.

Assessment of River Water Quality in an Agricultural Region of Brazil Using Biomarkers in a Native Neotropical Fish, Astyanax spp. (Characidae).

Intensive agricultural and livestock activities demand high pesticide use and, consequently, contaminants reach aquatic ecosystems. In the lower Jacuí River, southern Brazil, there is a lack of knowledge about pesticide residues in water samples and the biochemical responses in native fish species. Thus, this study aimed to estimate the influence of pesticide residues and water parameters to biomarker responses in the native fish Astyanax spp. We performed seasonal biomonitoring in 2017 with water samples and fish collections. Biomarkers of oxidative stress, antioxidants, biotransformation, and neurotoxicity were analyzed in fish tissues. Fourteen pesticide residues were detected; they presented correlations with detoxification enzyme and oxidative stress biomarkers. These data indicate that most of variations can be related to the pesticide presence in water indicating high aquatic pollution in this place.

Ecological risk of pesticide contamination in a Brazilian river located near a rural area: A study of biomarkers using zebrafish embryos.

Aquatic environments are affected by the use of pesticides in agricultural areas near rivers. To assess the impact of pesticide residues on affected environments Danio rerio (zebrafish) embryos have become an alternative model for biomonitoring studies. In the present study, zebrafish embryos were used as bioindicator of water quality in the Vacacaí river, located in the city of Santa Maria, southern Brazil. We hypothesized that it would be possible to observe changes in the biomarkers tested in the embryos. Exposures were performed over a total of eight months during the year 2018 using water collected in a river located near agricultural areas. Twenty-four pesticides were found in river water samples. The most frequently found were atrazine, quinclorac and clomazone. During exposure (96 h) spontaneous movement, the heart rate and hatching rate were evaluated. After the exposure time the embryos were euthanized for biochemical assays. We analyzed biomarkers such as thiobarbituric acid reactive substance (TBARS), acetylcholinesterase (AChE), glutathione S-transferase (GST) and catalase (CAT). We observed increases in GST and TBARS, especially during periods of major water contamination such as January, February, October, and November. Pesticides can affect the development of native species that reproduce during periods of high agricultural production. These results demonstrate the potential use of biochemical parameters combined with developmental and behavioral analyses in zebrafish embryos for biomonitoring studies.

Behavioural and biochemical parameters in guppy (Poecilia vivipara) following exposure to waterborne zinc in salt or hard water.

Zinc is an essential trace mineral that is involved in many biological processes. In elevated concentrations, this metal may have toxic effects for aquatic organisms. Physicochemical properties of water, such as salinity and hardness, can influence the bioavailability of zinc and, therefore its toxicity in aquatic environments. Therefore, this study aimed investigate the influence of salinity, hardness on Zn toxicity on the behaviours and biochemical parameters of the estuarine guppy (Poecilia vivipara). The fish were exposed to waterborne zinc (500 µg L-1) in salt water (25 ppt) or hard water (120 mg L-1 CaCO3). For behavioural analysis, the locomotive and exploratory parameters of fish in novel environment and light-dark tests were evaluated. We observed that exposure to hard water decreased the distance covered by the fish, and when zinc also present the vertical exploratory behavior decreased. When zinc was tested alone, an increase in the maximum speed of fish was recorded. Activities of antioxidant enzymes, levels of lipid peroxidation, protein carbonylation, total peroxidation and, reactive oxygen species content, antioxidant capacity against peroxyl radicals, non-proteins thiols levels, acetylcholinesterase and Na+/K+-ATPase activities were evaluated in the whole fish body. The integrated biomarker response was calculated for each parameter to aid in the interpretation of the results and indicated that hard water containing zinc had the greatest effect on the biochemical parameters of the fish. In general, neither salinity nor hardness were totally effective in protecting the guppy from the biochemical damage caused by exposure to zinc.

Zebrafish exposure to diphenyl diselenide-loaded polymeric nanocapsules caused no behavioral impairments and brain oxidative stress.

Previous findings showed that the nanoencapsulation of diphenyl diselenide [(PhSe)2], an organoselenium compound, provided superior biological effects and lower toxicological potential than its free form in vitro. However, few studies reported the behavioral and biochemical effects of this nanocapsules formulation in vivo. Zebrafish (Danio rerio) has emerged as a useful animal model to determine the pharmacological and toxicological effects of nanoparticles. Here, we evaluated the behavioral and brain oxidative effects after zebrafish exposure to (PhSe)2-loaded nanocapsules. Formulations were prepared by interfacial deposition of preformed polymer method and later tested at concentrations ranging from 0.1 to 2.0 µM. Both locomotor and exploratory activities were assessed in the novel tank diving test. Moreover, brain oxidative status was determined by measuring thiobarbituric acid-reactive substance levels, glutathione peroxidase, glutathione redutase and glutathione S-transferase activities. (PhSe)2-loaded nanocapsules showed no alteration on travelled distance, immobility, and erratic swimming, suggesting the absence of behavioral impairments. Interestingly, the higher concentration tested had anxiolytic-like effects, since animals spent more time in the top area and showed a decreased thigmotaxis behavior. Biochemical analysis demonstrated that the concentrations used in this study did not affect oxidative stress-related parameters in brain samples, reinforcing the low toxicological potential of the formulation. In conclusion, the exposure to (PhSe)2-loaded nanocapsules caused no locomotor impairments as well as did not modify the oxidative status of zebrafish brain, indicating that this formulation is probably non-toxic and promising for future pharmacological studies.