The influence of acute dopamine transporter inhibition on manic-, depressive-like phenotypes, and brain oxidative status in adult zebrafish.

Functional changes in dopamine transporter (DAT) are related to various psychiatric conditions, including bipolar disorder (BD) symptoms. In experimental research, the inhibition of DAT induces behavioral alterations that recapitulate symptoms found in BD patients, including mania and depressive mood. Thus, developing novel animal models that mimic BD-related conditions by pharmacologically modulating the dopaminergic signaling is relevant. The zebrafish (Danio rerio) has been considered a suitable vertebrate system for modeling BD-like responses, due to the well-characterized behavioral responses and evolutionarily conservation of the dopaminergic system of this species. Here, we investigate whether GBR 12909, a selective inhibitor of DAT, causes neurobehavioral alterations in zebrafish similar to those observed in BD patients. Behaviors were recorded after a single intraperitoneal (i.p.) administration of GBR 12909 at different doses (3.75, 7.5, 15 and 30 mg/kg). To observe temporal effects on behavior, swim path parameters were measured immediately after the administration period during 30 min. Locomotion, anxiety-like behavior, social preference, aggression, despair-like behavior, and oxidative stress-related biomarkers in the brain were measured 30 min post administration. GBR 12909 induced prominent effects on locomotor activity and vertical exploration during the 30-min period. Hyperactivity was observed in GBR 30 group after 25 min, while all doses markedly reduced vertical drifts. GBR 12909 elicited hyperlocomotion, anxiety-like behavior, decreased social preference, aggression, and induced depressive-like behavior in a behavioral despair task. Depending on the dose, GBR 12909 also decreased SOD activity and TBARS levels, as well as increased GR activity and NPSH content. Collectively, our novel findings show that a single GBR 12909 administration evokes neurobehavioral changes that recapitulate manic- and depressive-like states observed in rodents, fostering the use of zebrafish models to explore BD-like responses in translational neuroscience research.

Taurine modulates behavioral effects of intermittent ethanol exposure without changing brain monoamine oxidase activity in zebrafish: Attenuation of shoal- and anxiety-like responses, and abolishment of memory acquisition deficit.

Prolonged alcohol consumption has been considered as an important risk factor for various diseases. Chronic ethanol (EtOH) intake is associated with deleterious effects on brain functions culminating in robust behavioral changes. Notably, drugs available to treat the effects of EtOH have low therapeutic efficacy so far. Taurine (TAU) appears as a promising neuroprotective molecule due to its pleiotropic action in the brain. Here, we investigated whether TAU plays a beneficial role in different behavioral domains of zebrafish submitted to an intermittent EtOH exposure model, specially focusing on social behavior, anxiety-like responses, and memory. Moreover, since monoamines play a role in EtOH-mediated responses, we also evaluated the influence of both TAU and EtOH exposures on brain monoamine oxidase (Z-MAO) activity. Fish were exposed to non-chlorinated water or 1% EtOH for 8 consecutive days (20 min per day). From the 5th day until the end of the experimental period (8th day), animals were kept in the absence or presence of TAU (42, 150, or 400 mg/L) 1 h per day immediately after EtOH exposure. Behavioral measurements started 24 h after the last EtOH exposure. We observed that TAU showed modest attenuating effects on shoaling behavior and anxiety-like responses, while 42 and 150 mg/L TAU abolished the memory acquisition deficit in the inhibitory avoidance task. Biochemical analysis revealed that TAU did not modulate EtOH-induced increase on brain Z-MAO activity. Collectively, our novel data show a potential beneficial effect of TAU in an intermittent EtOH exposure model in zebrafish. Moreover, these findings foster the growing utility of this aquatic species to investigate the neurobehavioral basis of EtOH- and TAU-mediated responses in vertebrates.

Cypermethrin- and fipronil-based insecticides cause biochemical changes in Physalaemus gracilis tadpoles.

Insecticides used for agricultural pest control, as cypermethrin-based insecticide (CBI) and fipronil-based insecticide (FBI), are constant threats to non-target aquatic organisms. This study aimed to investigate the effect of different concentrations of cypermethrin and fipronil on neurotoxicity and oxidative stress in Physalaemus gracilis. Physalaemus gracilis tadpoles were exposed to five insecticide concentrations and a control treatment, with six replicates. During the experimental period, the tadpole mortality rate was evaluated and after 168 h, the neurotoxic enzyme activity and metabolite quantification related to the antioxidant system were measured. Tadpoles reduced acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities when exposed to 20 µg L-1 CBI and at all FBI concentrations, respectively. Glutathione S-transferase (GST) and superoxide dismutase (SOD) activities showed an increase from concentrations of 6 µg L-1 and 20 µg L-1 of CBI, respectively. After exposure of P. gracilis tadpoles to FBI, inhibitions of AChE and BChE were observed at the highest concentrations evaluated (500 and 1500 µg L-1). SOD activity decreased from 50 µg L-1 of FBI; however, catalase (CAT) and GST activities and carbonyl protein levels increased, regardless of the evaluated dose. We observed that both insecticides promoted oxidative stress and neurotoxic effects in P. gracilis tadpoles. These results suggest that biochemical biomarkers can be used for monitoring toxicity insecticides for the purpose of preservation of P. gracilis.

Impacts of dose and length of exposure to boldenone and stanazolol on enzymatic antioxidant systems, myeloperoxidase and NAGase activities, and glycogen and lactate levels in rat liver.

We investigated the adverse effects of the anabolic androgenic steroids (AAS) boldenone (BOL) and stanazolol (ST) on the enzymatic antioxidant systems of the rat liver. Male Wistar rats were divided in three protocols (P): PI, 5 mg/kg BOL or ST once a week for 4 weeks; PII, 2.5 mg/kg BOL or ST once a week for 8 weeks; PIII, 1.25 mg/kg BOL or ST once a week for 12 weeks. AAS were administered intramuscularly (0.2 ml, olive oil vehicle) once a week in all protocols. Activities of the enzymes glutathione peroxidase (GPx), glutathione S-transferase (GST), and glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT), were investigated. We assessed the content of hydrogen peroxide (H2O2), glycogen and lactate; and enzyme markers of neutrophils (myeloperoxidase, MPO) and macrophages (NAGase). PI and PII altered the SOD and CAT activities and increased the H2O2 content. PI led to increases in the MPO and NAGase activities. In contrast, changes in GPx, GST and, GR were observed under PII and, to a greater extend, under PIII. Following PIII, GPx, GR, and GST exhibited reduced activities. All protocols altered the glycogen and lactate content. The use of high doses of AAS for a short duration first alters SOD/CAT activity. In contrast, at lower doses of AAS for long periods is associated with changes in the glutathione system. Protocols with high doses of AAS for a short duration exert the most deleterious effects on redox status, markers of cellular infiltration, and the metabolic functioning of hepatic tissues.

Morphological and biochemical traits and mortality in Physalaemus gracilis (Anura: Leptodactylidae) tadpoles exposed to the insecticide chlorpyrifos.

Organophosphate insecticides such as chlorpyrifos are commonly detected in surface waters around the world, where they are highly toxic to many organisms. The frog Physalaemus gracilis uses water sources located in open fields as reproductive sites, where it is exposed to insecticides. The study aimed to evaluate the lethal and sublethal effect of a commercial chlorpyrifos formulation on P. gracilis tadpoles (Anura: Leptodactylidae). In acute toxicity tests, five chlorpyrifos concentrations between 750 and 2,000 µg L-1 were tested. Chronic toxicity, swimming activity, morphological and enzymatic changes, as well as levels of non-protein thiols (NPSH), carbonyl proteins and lipid peroxidation were evaluated at five insecticide concentrations between 11 and 500 µg L-1. The highest mortality rate of P. gracilis tadpoles occurred at 24 and 48 h, with an LC50 of 893.59 µg L-1. At all chlorpyrifos concentrations, tadpoles displayed reduced mobility and spasms. Morphological anomalies were observed in the mouth and intestine, especially at the highest concentrations used. Acetylcholinesterase activity decreased at 250 and 500 µg L-1, catalase activity increased at all concentrations, and superoxide dismutase and glutathione S-transferase increased from 90 µg L-1 to 30 µg L-1, respectively. We also observed increases in NPSH levels at chlorpyrifos concentration starting at 30 µg L-1 and increases in carbonyl proteins from 90 µg L-1 of pesticide. Taken together, these data suggest that the insecticide chlorpyrifos presents acute and chronic risks for P. gracilis, causing neurotoxic effects and oxidative damage, culminating in high risk for this species.

Hyperlipidemia-induced lipotoxicity and immune activation in rats are prevented by curcumin and rutin.

We assessed the effects of curcumin, rutin, and the association of rutin and curcumin in organs of hyperlipidemic rats. Rutin and curcumin have notable antioxidant and anti-inflammatory actions, so we hypothesized that their association would enhance their beneficial effects. Hyperlipidemia results in lipotoxicity and affects several organs. Lipotoxicity is not only an outcome of lipid accumulation in non-adipose tissues but also a result of the hyperlipidemia-associated inflammation and oxidative stress. Wistar rats were treated with rutin and curcumin for 30 days before the induction of acute hyperlipidemia by Poloxamer-407. After 36 h, the animals were euthanized for collection of blood and organs. Untreated hyperlipidemic rats showed higher uric acid and albumin levels in the serum and increased spleen size and ADA activity. Rutin, curcumin and the association reduced the spleen size by 20% and ADA activity by 23, 28, and 27%, respectively. Rats pretreated with rutin showed reduced lipid damage in the liver (40%) and the kidney (44%), and the protein damage was also reduced in the liver (75%). The lipid damage was decreased by 40% in the liver, and 56% in the kidney of rats pretreated with curcumin. The association reduced lipid damage by 50% and 36%, and protein damage by 77% and 64% in the liver and kidney, respectively. Rutin better prevented the decrease in the antioxidant defenses, increasing SOD by 34%, CAT by 246% and GST by 84% in the liver, as well as SOD by 119% and GST by 190% in the kidney. Also, analyses of blood and spleen parameters of untreated and pretreated non-hyperlipidemic rats showed no signs of immunotoxicity. Despite showing protective effects, the association did not perform better than the isolated compounds. Here, we showed that rutin and/or curcumin reestablished the immune homeostasis and redox balance disrupted by hyperlipidemia in peripheral organs of rats.

Involvement of anxiety-like behaviors and brain oxidative stress in the chronic effects of alarm reaction in zebrafish populations.

Aversive conditions elicit anxiety responses that prepare the organism to an eventual threat. Nonetheless, prolonged anxiety is a pathological condition associated with various neuropsychiatric disorders. Here, we evaluated whether the conspecific alarm substance (CAS), a chemical cue that elicits aversion, influences anxiety-like behaviors and modulates brain oxidative stress-related parameters in wild-type (WT) and leopard (leo) zebrafish following a repeated exposure protocol. CAS exposure was performed for 5 min, once daily for 7 consecutive days. In the 8th day, animals were tested in the light/dark and novel tank tests and their brains were further dissected for biochemical analyses. CAS chronically induced anxiogenic-like states in WT and leo populations when their behaviors were analyzed in the light/dark and novel tank tests. CAS also increased catalase (CAT) and glutathione S-transferase (GST) activities, as well as non-protein thiol (NPSH) content in WT and leo, but only leo had increased thiobarbituric reactive substance (TBARS) levels in the brain. At baseline conditions, leo was more 'anxious' when compared to WT, displaying lower CAT activity and carbonylated protein (CP) levels. Overall, CAS chronically triggers anxiety-like behavior in zebrafish populations, which may be associated with changes in oxidative stress-related parameters. Furthermore, the use of different zebrafish populations may serve as an interesting tool in future research aiming to investigate the neurobehavioral bases of neuropsychiatric disorders in vertebrates.

Taurine modulates the stress response in zebrafish.

The zebrafish (Danio rerio) is used as an emergent model organism to investigate the behavioral and physiological responses to stress. The anxiolytic-like effects of taurine in zebrafish support the existence of different mechanisms of action, which can play a role in preventing stress-related disorders (i.e., modulation of GABAA, strychnine-sensitive glycine, and NMDA receptors, as well as antioxidant properties). Herein, we investigate whether taurine modulates some behavioral and biochemical responses in zebrafish acutely submitted to chemical and mechanical stressors. We pretreated zebrafish for 1 h in beakers at 42, 150, and 400 mg/L taurine. Fish were later acutely exposed to a chemical stressor (conspecific alarm substance) or to a mechanical stressor (net chasing), which elicits escaping responses and aversive behaviors. Locomotion, exploration, and defensive-like behaviors were measured using the novel tank and the light-dark tests. Biochemical (brain oxidative stress-related parameters) and whole-body cortisol levels were also quantified. We showed that taurine prevents anxiety/fear-like behaviors and protein carbonylation and dampens the cortisol response following acute stress in zebrafish. In summary, our results demonstrate a protective role of taurine against stress-induced behavioral and biochemical changes, thereby reinforcing the growing utility of zebrafish models to investigate the neuroprotective actions of taurine in vertebrates.

Taurine Protects from Pentylenetetrazole-Induced Behavioral and Neurochemical Changes in Zebrafish.

Epilepsy is a common neurological disorder characterized by recurrent unprovoked seizures, which culminate in various neurobehavioral and neurochemical changes. Taurine (TAU) is an amino sulfonic acid which acts an endogenous inhibitory neuromodulator. Moreover, TAU displays intrinsic antioxidant activity, contributing to its beneficial actions in the CNS. Here, we evaluated whether TAU pretreatment protects from pentylenetetrazole (PTZ)-induced behavioral alterations and oxidative stress-related parameters in zebrafish brain tissue. Fish were pretreated with 42, 150, and 400 mg/L TAU (40 min) and further exposed to 10 mM PTZ (20 min) to analyze the seizure-like behaviors. As a positive control, another group was previously treated with 75 µM diazepam (DZP). Afterwards, biochemical experiments were performed. All TAU concentrations tested decreased seizure intensity in the first 150 s. Importantly, 150 mg/L TAU attenuated seizure-like behavioral scores, decreased seizure intensity, reduced the frequency of clonic-like seizures (score 4), and increased the latency to score 4. TAU (150 mg/L) also prevented oxidative stress in PTZ-challenged fish by decreasing lipid peroxidation and protein carbonylation and preventing changes on nonprotein thiol levels. No significant changes were observed in MTT assay and LDH activity. Differently than observed in DZP group, TAU did not affect the overall swimming activity of fish, suggesting different mechanisms of action. Collectively, we show that TAU attenuates PTZ-induced seizure-like behaviors and brain oxidative stress in zebrafish, suggesting the involvement of antioxidant mechanisms in neuroprotection.

Hyperglycemia elicits anxiety-like behaviors in zebrafish: Protective role of dietary diphenyl diselenide.

Diabetes mellitus (DM) is a chronic metabolic disease that may comorbid with various psychiatric disorders, such as anxiety and depression. The search for effective therapeutics to alleviate hyperglycemia and complications resulting from DM is continuous. Here we investigate the effects of diphenyl diselenide (DD), an organoselenium compound with several pharmacological properties, in a zebrafish model of hyperglycemia. Fish were fed for 74 days with a diet containing 3 mg/Kg DD, a concentration chosen after experiments based in a dose-response curve (DD 1, 2 and 3 mg/Kg) that did not cause overt toxicity (mortality, weight loss and neurobehavioral deficits). In the last 14 days of the experimental period, fish were concomitantly exposed to a glucose solution (111 mM). Afterwards, blood glucose levels, brain selenium (Se) content, and behavioral analysis aiming to assess anxiety-like behaviors and locomotor/exploratory activities were performed. In the novel tank diving test, glucose decreased vertical exploration and fish spent less time in the lit area when tested in the light-dark test, suggesting increased anxiety-like behavior. Moreover, DD decreased blood glucose levels in hyperglycemic fish as well as prevented the development of anxiety-related symptoms. DD diet alone did not change glycemia and behavioral parameters, but increased Se levels in the brain without affecting the cellular viability. Collectively, our findings highlight the growing utility of this zebrafish hyperglycemia model as a valuable strategy for further research in DM field and neuroprotective approaches.

Oxidative effects of the acute exposure to a pesticide mixture of cypermethrin and chlorpyrifos on carp and zebrafish - A comparative study.

The use of commercial pesticides combinations increases the risk of intoxication in non-target aquatic organisms. Here, we investigate the potential of a commercial pesticide formulation containing (CYP) plus chlorpyrifos (CPF) to induce oxidative damage on two fish species (common carp and zebrafish). Carp and zebrafish were exposed for 96 h under laboratory conditions. Fish were divided in three different groups: CTL, 0.3 µg L-1 or 0.6 µg L-1 of CYP and 0.5 or 1 µg L-1 of CPF in commercial formulation. Both carp and zebrafish showed an increase in lipid peroxidation (LPO) and glutathione-S-transferase (GST) activity when compared to control group. Other oxidative parameters responded differently to exposure in carp and zebrafish. There were an increase in ascorbic acid (ASA) levels and decrease in catalase (CAT) activity and non-protein thiols (NPSH) levels in treated groups of carps. In the other hand, zebrafish showed significant decrease in ASA and increase in CAT activity and NPSH levels. Overall, we demonstrate noxious effects on redox parameters in two fish experimental models and different effects were observe in each fish species exposed to commercial pesticide formulation. This difference responses observed can be related with specific mechanisms of detoxification and antioxidant defense system of each species.

Bioaccumulation and oxidative stress caused by pesticides in Cyprinus carpio reared in a rice-fish system.

Nowadays, many irrigated rice producers have adopted the rice-fish farming systems, in which fish are reared in the same environment as rice is grown. These animals can biologically control many pests that disturb the crop, as well as increase the income of producers when they are reared in the same area as rice. However, a large variety of pesticides is often used for pest control in conventional irrigated rice crops, and fish are exposed to different pesticide concentrations and active ingredients. Thus, the aim of the current study is to assess the potential risks of these pesticides to freshwater fish (Cyprinus carpio) cultured in integrated rice-fish farming systems. We assessed five pesticides widely used in rice culture. This is the first report on the bioaccumulation of the insecticides lambda-cyhalothrin + thiamethoxam and clorantraniliprole and of the fungicide tebuconazole + trifloxystrobin associated with oxidative stress and with enzymatic and non-enzymatic parameters in edible fish samples reared in integrated in rice-fish farming systems. After 100 days of exposure, lambda-cyhalothrin and tebuconazole bioaccumulated in carp muscles. Acethylcholinesterase activity in the brain was lower in the experimental group than in the control group. Overall, the biochemical analyses showed increased responses in the experimental groups in comparison to the controls. Lipid peroxidation levels increased in the liver, gills and muscles. Protein oxidation increased in the liver and muscles. Catalase activity increased in the liver, and glutathione S-transferase increased in the liver, gills and muscles. Results demonstrate that these pesticides have severe adverse consequences in fish and indicate their potential risk to human health due to their bioaccumulation in farmed fish.

Sodium Selenite Prevents Paraquat-Induced Neurotoxicity in Zebrafish.

Considering the antioxidant properties of sodium selenite (Na2SeO3) and the involvement of oxidative stress events in paraquat-induced neurotoxicity, this study investigated the protective effect of dietary Na2SeO3 on biochemical and behavioral parameters of zebrafish exposed to paraquat (PQ). Fish were pretreated with a Na2SeO3 diet for 21 days and then PQ (20 mg/kg) was administered intraperitoneally with six injections for 16 days. In the novel tank test, the Na2SeO3 diet prevented the locomotor impairments, as well as the increase in the time spent in the top area of the tank, and the exacerbation of freezing episodes. In the preference for conspecifics and in the mirror-induced aggression (MIA) tasks, Na2SeO3 prevented the increase in the latency to enter the area closer to conspecifics and the agonistic behavior of PQ-treated animals, respectively. Na2SeO3 prevented the increase of carbonylated protein (CP), reactive oxygen species (ROS), and nitrite/nitrate (NOx) levels, as well as the decrease in non-protein thiols (NPSH) levels. Regarding the antioxidant enzymatic defenses, Na2SeO3 prevented the increase in catalase (CAT) and glutathione peroxidase (GPx) activities caused by PQ. Altogether, dietary Na2SeO3 improves behavioral and biochemical function impaired by PQ treatment in zebrafish, by modulating not only redox parameters, but also anxiety- and aggressive-like phenotypes in zebrafish.

Triphenyltin hydroxide induces changes in the oxidative stress parameters of fish.

Among all organotin compounds , triphenyltin hydroxide (TPhTH) is widely used as fungicide and moluscicide in Brazil. However, the effects of TPhTH on the biochemical parameters of non-target organisms, such as fish, are little known. The aim of the present study is to assess the possible toxic effects of different concentrations of waterborne TPhTH on silver catfish belonging to species Rhamdia quelen. The fish were exposed to two different concentrations of TPhTH (1.08 and 1.70 µg/L as Sn) for 15 days and then compared to the control group (triplicate, n = 3). The antioxidant profile (catalase (CAT) and the glutathione S-transferase (GST)) and the oxidative stress parameters (TBARS-thiobarbituric acid-reactive substances and protein carbonyl (PC)) were set after the exposure to TPhTH. The TBARS level and the PC content increased in several organs of the Rhamdia quelen (brain, liver, muscle and gills) under the two concentrations of TPhTH in comparison to the control group. The CAT activity in the liver and gills has enhanced in all tested TPhTH concentrations. The GST activity increased in the brain, liver and muscle tissues under all the TPhTH concentrations. The significant changes in the biomarkers indicated that the investigated pesticide could have harmful effect on fish, in the field. However, these biomarkers were measured after the fish received doses lower than the recommended for use in agriculture.

Quercetin changes purinergic enzyme activities and oxidative profile in platelets of rats with hypothyroidism.

Diseases related to thyroid hormones have been extensively studied because affect a large number of individuals, and these hormones participate in the regulation of the whole organism homeostasis. However, little is known about the involvement of purinergic signaling related to oxidative stress in hypothyroidism and possible therapeutic adjuncts for treatment of this disorder. Thus, the present study investigates the effects of quercetin on NTPDase, 5'-nucleotidase and adenosine deaminase activities, platelet aggregation and oxidative profile in platelets of rats with methimazole (MMI)-induced hypothyroidism. Methimazole at a concentration of 20mg/100mL was administered for 90days. From the second month the animals received quercetin 10 or 25mg/kg for 60days. Results showed that: Ecto-5'-nucleotidase activity decreased in methimazole/water group and the treatment with quercetin 25mg/kg decreased NTPDase, 5'-nucleotidase and adenosine deaminase activities. Moreover, platelet aggregation increased in methimazole/water group. Lipid peroxidation increased while superoxide dismutase and catalase activities decreased, but, interestingly, the treatment with quercetin reversed these changes. These results demonstrated that quercetin modulates adenine nucleotide hydrolysis decreasing the ADP formation and adenosine deamination. At the same time quercetin improves the oxidative profile, as well as reduces platelet aggregation, which together with the modulation in the nucleotides levels can contribute to the prevention of platelet disorders.

Pre-sedation and transport of Rhamdia quelen in water containing essential oil of Lippia alba: metabolic and physiological responses.

The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 µL/L of essential oil (EO) from Lippia alba leaves were investigated. Prior to transport, the fish in the two experimental groups were sedated with 200 µL/L of EO for 3 min. After transport, dissolved oxygen, carbon dioxide, alkalinity, water hardness, pH, temperature and un-ionized ammonia levels in the transport water did not differ significantly among the groups. However, total ammonia nitrogen levels and net Na(+), Cl(-) and K(+) effluxes were significantly lower in the groups transported with EO of L. alba than those in the control group. PvO2, PvCO2 and HCO3(-) were higher after transporting fish in 40 µL/L of EO of L. alba, but there were no significant differences between groups regarding blood pH or hematocrit. Cortisol levels were significantly higher in fish transported in 30 µL/L of EO of L. alba compared to those of the control group. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) showed different responses after adding EO to the transport water. In conclusion, while the EO of L. alba is recommended for fish transport in the conditions tested in the present study because it was effective in reducing waterborne total ammonia levels and net ion loss, the higher hepatic oxidative stress in this species with the same EO concentrations reported by a previous study led us to conclude that the 10-20 µL/L concentration range of EO and lack of pre-sedation before transport are more effective.

Exposure to different glyphosate formulations on the oxidative and histological status of Rhamdia quelen.

Due to the wide use of glyphosate (GLY) in soybean cultivation, their residues in the environment may affect non-target organisms such as fish, developing toxic effects. Despite GLY being widely used in Brazil, there are few studies comparing the effects of commercial formulations in native freshwater fish species. Silver catfish (Rhamdia quelen) were exposed to three different commercial formulations of GLY 48% (Orium(®), Original(®) and Biocarb(®)) at 0.0, 2.5 and 5.0 mg/L for 96 h. The effects in thiobarbituric acid-reactive substances (TBARS), catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) and histological alterations were analysed in the liver, whereas alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were studied in the plasma. In the liver, TBARS levels increased and CAT decreased in all treatments and herbicides tested in comparison with the control group. The SOD increased at 2.5 mg/L of Orium(®), Original(®) and 5.0 mg/L Orium(®) and Biocarb(®), whereas GST increased at 2.5 mg/L Orium(®) and decreased at 2.5 mg/L Biocarb(®) when compared to the control group. The main histopathological alterations in hepatic tissue were vacuolisation, leucocyte infiltration, degeneration of cytoplasm and melanomacrophage in all GLY treatments. The ALT decreased after exposure to 2.5 mg/L of Biocarb(®) and AST increased at 2.5 mg/L of Orium(®), Original(®) and 5.0 mg/L of Biocarb(®) in comparison with the control group. In summary, the oxidative damage generated by GLY may have caused the increased formation of free radicals that led to the histological alterations observed in hepatocytes.

Strain- and context-dependent behavioural responses of acute alarm substance exposure in zebrafish.

We investigate the behavioural responses of wild type (WT) and leopard (leo) zebrafish elicited by alarm substances of conspecifics at three contexts: during the exposure period (Experiment 1); after exposure, in habituation to novelty (Experiment 2); or after exposure, in the light-dark preference test (Experiment 3), and analyse their influence on pigment response. During the exposure, leo showed decreased vertical drifts, increased number and duration of erratic movements, while WT had increased erratic movements and latency to enter the top. In the novel tank, we observed that angular velocity decreased in WT exposed to alarm substance, which also presented increased fear responses. Contrastingly, leo increased the number of entries and time in top, indicating differences in habituation profile. Alarm substance increased the number of erratic movements in the light-dark test, but elicited different responses between strains in scototaxis, latency to enter the dark compartment and risk assessment episodes. Moreover, the body colour of zebrafish did not change after alarm substance exposure. Principal component analyses suggest that burst swimming, anxiety-like behaviours, and locomotion/exploration were the components that most accounted for total variances of Experiments 1, 2, and 3, respectively. We conclude that chemical cue from conspecifics triggers strain- and context-dependent responses.

Azadirachtin, a neem-derived biopesticide, impairs behavioral and hematological parameters in carp (Cyprinus carpio).

Azadirachtin (Aza) is a promisor biopesticide used in organic production and aquaculture. Although this compound is apparently safe, there is evidence that it may have deleterious effects on fish. Behavioral and hematological tests are grouped into a set of parameters that may predict potential toxicity of chemical compounds. Here, we investigate the effects of Aza, in the commercial formulation Neenmax™ , on carp (Cyprinus carpio) by defining LC50 (96 h), and testing behavioral and hematological parameters. In our study, LC50 was estimated at 80 µL/L. We exposed carp to Aza at 20, 40, and 60 µL/L, values based on 25, 50, and 75% of LC50 , respectively. At 60 µL/L, Aza promoted significant changes in several parameters, increasing the distance traveled and absolute turn angle. In addition, the same concentration decreased the time spent immobile and the number of immobile episodes. Hematological parameters, such as hematocrit, hemoglobin, hematimetrics index, and red cell distribution, were decreased at 60 µL/L Aza exposure. In conclusion, our study demonstrates that 60 µL/L Aza altered locomotor activity, motor pattern, and hematological parameters, suggesting potential toxicity to carp after acute exposure. In addition, this is the first report that evaluates the actions of a chemical contaminant using automated behavioral tracking of carp, which may be a useful tool for assessing the potential toxicity of biopesticides in conjunction with hematological tests. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1381-1388, 2016.

Acute exposure to the biopesticide azadirachtin affects parameters in the gills of common carp (Cyprinus carpio).

The biopesticide, azadirachtin (Aza) is less hazardous to the environment, but may cause several toxic effects in aquatic organisms. The Cyprinus carpio (n=12, for all concentrations) after 10days of acclimation under controlled conditions, were exposed at 20, 40, and 60µL/L of Aza during 96h. After this period, fish were anesthetized and euthanized then mucus layer and gills collected. In this study, the effects of exposure to different Aza concentrations were analysed through a set of biomarkers: Na(+)/K(+-)ATPase, lipid peroxidation (TBARS), protein carbonyl (PC), superoxide dismutase (SOD), glutathione-S-transferase (GST), catalase (CAT), glutathione peroxidase (GPx), non-protein thiols (NPSH), ascorbic acid (AsA) and histological parameters and, yet, protein and glucose concentration in the surface area of mucous layer. Na(+)K(+-)ATPase was inhibited at 40 and 60µL/L compared to control. TBARS decreased at 40µL/L compared to control. PC, SOD and GST increased at 60µL/L in comparison to control. CAT increased at 20 and 60µL/L, and GPx increased in all Aza concentrations compared to control. NPSH decreased and AsA increased in all concentrations in comparison to control. Histological analyses demonstrated an increase in the intensity of the damage with increasing Aza concentration. Alterations in histological examination were elevation and hypertrophy of the epithelial cells of the secondary filament, hypertrophy and hyperplasia of the mucous and chlorate cells and lamellar aneurism. Glucose and protein concentrations in mucus layer increased at 60µL/L compared to control. In general, we suggest that 60µL/L Aza concentration affected several parameters causing disruptions carp metabolism.