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.

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.

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.

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.

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.

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.

Anesthetic activity and bio-guided fractionation of the essential oil of Aloysia gratissima (Gillies & Hook.) Tronc. in silver catfish Rhamdia quelen.

This work aimed to determine the efficacy of the essential oil of A. gratissima as anesthetic for silver catfish, and to perform the bio-guided fractionation of essential oil aiming to isolate compounds responsible for the noted effects. Fish were submitted to anesthesia bath with essential oil, its fractions and isolated compounds to determine time of anesthetic induction and recovery. Eugenol (50 mg L(-1)) was used as positive control. Essential oil of A. gratissima was effective as an anesthetic at concentrations of 300 to 900 mg L(-1). Fish presented involuntary muscle contractions during induction and recovery. The bio-guided fractionation of essential oil furnished E-(-)-pinocamphone, (-)-caryophyllene oxide, (-)-guaiol and (+)-spathulenol. E-(-)-pinocamphone caused the same side effects observed for essential oil. (-)-Caryophyllene oxide, (-)-guaiol and (+)-spathulenol showed only sedative effects at proportional concentrations to those of the constituents in essential oil. (+)-Spathulenol (51.2 mg L(-1)) promoted deep anesthesia without side effects. A higher concentration of (+)-spathulenol, and lower or absent amounts ofE-(-)-pinocamphone could contribute to increase the activity and safety of the essential oil of A. gratissima. (+)-Spathulenol showed potent sedative and anesthetic activities in silver catfish, and could be considered as a viable compound for the development of a new anesthetic.

The essential oil from Lippia alba induces biochemical stress in the silver catfish (Rhamdia quelen) after transportation

This study investigated the effects of the essential oil (EO) from Lippia alba on biochemical parameters related to oxidative stress in the brain and liver of silver catfish (Rhamdia quelen) after six hours of transport. Fish were transported in plastic bags and divided into three treatments groups: control, 30 µL L- 1 EO from L.alba and 40 µL L-1 EO from L.alba. Prior to transport, the fish were treated with the EO from L. alba (200 µL L -1 for three minutes), except for the control group. Fish transported in bags containing the EO did not have any alterations in acetylcholinesterase, ecto -nucleoside triphosphate diphosphohydrolase and 5'nucleotidase activity in the brain or superoxide dismutase activity in the liver. The hepatic catalase (CAT), glutathione-S-transferase (GST), glutathione peroxidase (GPx), nonprotein thiol and ascorbic acid levels were significantly lower compared to the control group. However, the hepatic thiobarbituric acid- reactive substances, protein oxidation levels and the lipid peroxidation/catalase+glutathione peroxidase (LPO/CAT+GPx) ratio were significantly higher in fish transported with both concentrations of the EO, indicating oxidative stress in the liver. In conclusion, considering the hepatic oxidative stress parameters analyzed in the present experiment, the transport of previously sedated silver catfish in water containing 30 or 40 µL L-1 of EO from L. alba is less effective than the use of lower concentrations.

Este estudo investigou os efeitos do óleo essencial (OE) de Lippia alba sobre parâmetros bioquímicos relacionados ao estresse oxidativo em cérebro e fígado de jundiá (Rhamdia quelen), após seis horas de transporte. Os peixes foram transportados em sacos plásticos e divididos em três tratamentos: controle, 30 µL L-1 e 40 µL L-1 de OE de L.alba. Antes do transporte, os peixes foram tratados com o OE de L. alba (200 µL L-1 por três minutos), exceto para o grupo controle. Os peixes transportados em sacos contendo o OE não tiveram alterações na atividade da acetilcolinesterase (AChE), ecto-nucleosídeo trifosfato difosfohidrolase (NTPDase) e 5'nucleotidase, em cérebro ou superóxido dismutase (SOD) no fígado. O tiol não proteico (NPSH), os níveis de ácido ascórbico, catalase (CAT), glutationa-S-transferase (GST) e glutationa-peroxidase (GPx) hepáticos, foram significativamente mais baixos em comparação com o grupo controle. No entanto, as substâncias reativas ao ácido tiobarbitúrico (TBARS), os níveis de oxidação proteica e a taxa de peroxidação lipídica/catalase+glutationa peroxidase (LPO/ CAT+GPx) foram significativamente maiores nos peixes transportados com ambas as concentrações de OE, indicando estresse oxidativo no fígado. Em conclusão, considerando os parâmetros de estresse oxidativo do fígado analisados no presente experimento, o transporte de jundiás previamente sedados em água contendo 30 ou 40 µL L-1 de OE de L.alba é menos efetivo que utilizando concentrações menores.

The essential oil from Lippia alba induces biochemical stress in the silver catfish (Rhamdia quelen) after transportation

This study investigated the effects of the essential oil (EO) from Lippia alba on biochemical parameters related to oxidative stress in the brain and liver of silver catfish (Rhamdia quelen) after six hours of transport. Fish were transported in plastic bags and divided into three treatments groups: control, 30 µL L- 1 EO from L.alba and 40 µL L-1 EO from L.alba. Prior to transport, the fish were treated with the EO from L. alba (200 µL L -1 for three minutes), except for the control group. Fish transported in bags containing the EO did not have any alterations in acetylcholinesterase, ecto -nucleoside triphosphate diphosphohydrolase and 5'nucleotidase activity in the brain or superoxide dismutase activity in the liver. The hepatic catalase (CAT), glutathione-S-transferase (GST), glutathione peroxidase (GPx), nonprotein thiol and ascorbic acid levels were significantly lower compared to the control group. However, the hepatic thiobarbituric acid- reactive substances, protein oxidation levels and the lipid peroxidation/catalase+glutathione peroxidase (LPO/CAT+GPx) ratio were significantly higher in fish transported with both concentrations of the EO, indicating oxidative stress in the liver. In conclusion, considering the hepatic oxidative stress parameters analyzed in the present experiment, the transport of previously sedated silver catfish in water containing 30 or 40 µL L-1 of EO from L. alba is less effective than the use of lower concentrations.(AU)

Este estudo investigou os efeitos do óleo essencial (OE) de Lippia alba sobre parâmetros bioquímicos relacionados ao estresse oxidativo em cérebro e fígado de jundiá (Rhamdia quelen), após seis horas de transporte. Os peixes foram transportados em sacos plásticos e divididos em três tratamentos: controle, 30 µL L-1 e 40 µL L-1 de OE de L.alba. Antes do transporte, os peixes foram tratados com o OE de L. alba (200 µL L-1 por três minutos), exceto para o grupo controle. Os peixes transportados em sacos contendo o OE não tiveram alterações na atividade da acetilcolinesterase (AChE), ecto-nucleosídeo trifosfato difosfohidrolase (NTPDase) e 5'nucleotidase, em cérebro ou superóxido dismutase (SOD) no fígado. O tiol não proteico (NPSH), os níveis de ácido ascórbico, catalase (CAT), glutationa-S-transferase (GST) e glutationa-peroxidase (GPx) hepáticos, foram significativamente mais baixos em comparação com o grupo controle. No entanto, as substâncias reativas ao ácido tiobarbitúrico (TBARS), os níveis de oxidação proteica e a taxa de peroxidação lipídica/catalase+glutationa peroxidase (LPO/ CAT+GPx) foram significativamente maiores nos peixes transportados com ambas as concentrações de OE, indicando estresse oxidativo no fígado. Em conclusão, considerando os parâmetros de estresse oxidativo do fígado analisados no presente experimento, o transporte de jundiás previamente sedados em água contendo 30 ou 40 µL L-1 de OE de L.alba é menos efetivo que utilizando concentrações menores.(AU)

Toxicity of triphenyltin hydroxide to fish.

Triphenyltin (TPhT) is used worldwide in pesticide formulas for agriculture. Toxic effects of this compound to aquatic life have been reported; however, the biochemical response of fish exposed to different concentrations of TPhT hydroxide (TPhTH) was investigated for the first time in this study. The lethal concentration (LC50) of TPhTH to silver catfish, Rhamdia quelen, was calculated from an acute-exposure experiment (96 h). In addition, acethylcholinesterase (AChE) activity in brain and muscle-as well as glucose, glycogen, lactate, total protein, ammonia, and free amino acids in liver and muscle-were evaluated in a chronic-exposure experiment (15-day exposure). Speciation analysis of tin (Sn) was performed in fish tissues at the end of both experiments using gas chromatography coupled to a pulsed-flame photometric detector (GC-PFPD). Concentrations of TPhT, diphenyltin, and monophenyltin (reported as Sn) were lower than limits of quantification (10σ criteria). Waterborne TPhTH concentration used through the experiment was also evaluated by GC-PFPD, and no degradation of this species was observed. The LC50 value for silver catfish juveniles was 9.73 µg L(-1) (as Sn). Decreased brain and muscle AChE activities were observed in fish exposed to TPhTH in relation to unexposed fish (control). Liver glycogen and lactate levels were significantly higher in fish kept at the highest waterborne TPhTH concentration compared with the control. Liver and muscle glucose levels of fish exposed to all TPhTH concentrations were significantly lower than those of control fish. Silver catfish exposed to all TPhTH concentrations showed lower total protein values and higher total free amino acids levels in liver and muscle compared with controls. Total ammonia levels in liver and muscle were significantly higher for the highest TPhTH concentration compared with controls. In conclusion, TPhTH caused metabolic alterations in silver catfish juveniles, and the analyzed parameters can also be used as bioindicators for TPhTH contamination.

Mortality, bioaccumulation and physiological responses in juvenile freshwater mussels (Lampsilis siliquoidea) chronically exposed to copper.

Several studies have indicated that the early life stages of freshwater mussels are among the most sensitive aquatic organisms to inorganic chemicals, including copper. However, little is known about the toxic mode of action and sub-lethal effects of copper exposure in this group of imperiled animals. In this study, the physiological effects of long-term copper exposure (survival, growth, copper bioaccumulation, whole-body ion content, oxygen consumption, filtration rate, ATPase activities, and biomarkers of oxidative stress) were evaluated in juvenile (6 month old) mussels (Lampsilis siliquoidea). The mussels' recovery capacity and their ability to withstand further acute copper challenge were also evaluated in secondary experiments following the 28 day exposure by assessing survival, copper bioaccumulation and whole-body ion content. Mussels chronically exposed to 2 and 12 µg Cu/L showed significantly higher mortality than those held under control conditions (mortality 20.9, 69.9 and 12.5%, respectively), indicating that juvenile L. siliquoidea is underprotected by the U.S. Environmental Protection Agency (USEPA) biotic ligand model (BLM)-derived chronic water quality criteria (WQC) (2.18 µg Cu/L) and the hardness-derived USEPA WQC (12.16 µg Cu/L). Soft tissue copper burden increased equally for both copper exposures, suggesting that chronic toxicity is not associated with copper bioaccumulation. Several physiological disturbances were also observed during chronic copper exposure. Most relevant was a decrease in whole-body sodium content paralleled by an inhibition of Na(+) K(+)-ATPase activity, indicating a metal-induced ionoregulatory disturbance. Filtration and oxygen consumption rates were also affected. Redox parameters (reactive oxygen production, antioxidant capacity against peroxyl radicals, glutathione-S-transferase (GST) activity, and glutathione (GSH) concentration) did not show clear responses, but membrane damage as lipid peroxidation (LPO) was observed in both copper exposures. Mussels previously held in control conditions or pre-exposed to 2 µg dissolved Cu/L were able to maintain their ionic homeostasis and did not experience mortality after the 4-d recovery period. In contrast, those previously exposed to 12 µg dissolved Cu/L exhibited 50% mortality indicating that they had already reached a 'point of no return'. Pre-exposure to copper did not influence mussel response to the copper challenge test. As observed for the chronic exposure, mortality of mussels held in the absence of copper and submitted to the challenge test was also associated with an ionoregulatory disturbance. These results indicate that ionoregulatory disruption in freshwater mussels chronically exposed to copper is the main mechanism of toxicity and that redox parameters do not appear to be useful as indicators of sub-lethal copper toxicity in these animals.

Oxidative stress biomarkers and acetylcholinesterase activity in human erythrocytes exposed to clomazone (in vitro).

The aim of this study was to investigate the effect of clomazone herbicide on oxidative stress biomarkers and acetylcholinesterase activity in human erythrocytes in in vitro conditions. The activity of catalase (CAT), superoxide dismutase (SOD) and acetylcholinesterase (AChE), as well as the levels of thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH) were measured in human erythrocytes exposed (in vitro) to clomazone at varying concentrations in the range of 0, 100, 250 and 500 µg/L for 1 h at 37 °C.TBARS levels were significantly higher in erythrocytes incubated with clomazone at 100, 250 and 500 µg/L. However, erythrocyte CAT and AChE activities were decreased at all concentrations tested. SOD activity was increased only at 100 µg/L of clomazone. GSH levels did not change with clomazone exposure. These results clearly showed clomazone to induce oxidative stress and AChE inhibition in human erythrocytes (in vitro). We, thus, suggest a possible role of ROS on toxicity mechanism induced by clomazone in humans.

Survival, growth and metabolic parameters of silver catfish, Rhamdia quelen, juveniles exposed to different waterborne nitrite levels

Altos níveis de nitrito (NO2-) podem ocorrer em sistemas de cultivo com alta densidade de estocagem, mas análises sobre os valores de concentração letal e o efeito do NO2 - em parâmetros metabólicos e no crescimento são escassos. Neste estudo foi analisada a concentração letal em 96 h (CL50-96h) para nitrito (NO2 -) em juvenis de jundiá, Rhamdia quelen, e o efeito de quatro níveis de nitrito (0,06; 0,46; 1,19 e 1,52 mg.L-1) no crescimento e no lactato, glicose, glicogênio e proteína hepática e muscular. A CL50-96h para NO2 - foi 20,46 (intervalo de confiança: 16,10-23,68) mg.L-1. No experimento de crescimento, a exposição ao NO2 não afetou o peso, comprimento ou taxa de crescimento específico, mas devido à mortalidade (66,7% e 100% após 20 e 40 dias, respectivamente), a biomassa dos juvenis expostos a 1,52 mg.L-1. NO2 - foi significativamente mais baixa que a biomassa dos juvenis expostos aos outros tratamentos. Deste modo, o nível seguro de NO2 - para o crescimento do jundiá é abaixo de 1,19 mg.L-1 (2% da CL50-96h). A exposição do jundiá ao NO2 - por 40 dias diminuiu os níveis de lactato no músculo, mas esses níveis aumentaram nos exemplares mantidos em 1,19 mg.L-1 NO2 -. Além disso, os níveis de glicose no músculo e fígado foram significativamente mais baixos nos jundiás expostos à concentração mais elevada de NO2 . Estes resultados indicam que a exposição crônica ao NO2 provoca uma oxidação anaeróbica do substrato para obtenção de energia.

High nitrite (NO2 -) levels may develop in aquaculture systems due to high fish density, but studies of lethal concentration values and the effect of NO2 - on metabolic parameters and growth are scarce. Consequently, in this study was verified the lethal concentration at 96 h (LC50-96h) for (NO2 -) in juvenile silver catfish, Rhamdia quelen and the effect of four waterborne NO2 - concentrations (0.06, 0.46, 1.19, and 1.52 mg.L-1) on growth, and hepatic and muscular lactate, glucose, glycogen and protein. Nitrite LC50-96h was 20.46 (confidence interval: 16.10-23.68) mg.L-1. In the growth experiment, exposure to NO2 - did not affect weight, length or specific growth rate, but due to mortality (66.7% and 100% after 20 and 40 days, respectively), biomass of juveniles exposed to 1.52 mg.L-1 NO2 - was significantly lower than the biomass of juveniles exposed to other treatments. Therefore, the safe level of nitrite for growth of silver catfish juveniles is below 1.19 mg.L-1 (2% of LC50-96h). Exposure of silver catfish to NO2 - for 40 days reduced lactate levels in muscle, but lactate levels increased in liver tissue of fish maintained at 1.19 mg.L-1 NO2 -. In addition, glucose levels in muscle and liver tissues were significantly lower in silver catfish exposed to the highest NO2 - level. These results indicate that chronic NO2 - exposure causes anaerobic substrate oxidation to meet energy demand.

Survival, growth and metabolic parameters of silver catfish, Rhamdia quelen, juveniles exposed to different waterborne nitrite levels

Altos níveis de nitrito (NO2-) podem ocorrer em sistemas de cultivo com alta densidade de estocagem, mas análises sobre os valores de concentração letal e o efeito do NO2 - em parâmetros metabólicos e no crescimento são escassos. Neste estudo foi analisada a concentração letal em 96 h (CL50-96h) para nitrito (NO2 -) em juvenis de jundiá, Rhamdia quelen, e o efeito de quatro níveis de nitrito (0,06; 0,46; 1,19 e 1,52 mg.L-1) no crescimento e no lactato, glicose, glicogênio e proteína hepática e muscular. A CL50-96h para NO2 - foi 20,46 (intervalo de confiança: 16,10-23,68) mg.L-1. No experimento de crescimento, a exposição ao NO2 não afetou o peso, comprimento ou taxa de crescimento específico, mas devido à mortalidade (66,7% e 100% após 20 e 40 dias, respectivamente), a biomassa dos juvenis expostos a 1,52 mg.L-1. NO2 - foi significativamente mais baixa que a biomassa dos juvenis expostos aos outros tratamentos. Deste modo, o nível seguro de NO2 - para o crescimento do jundiá é abaixo de 1,19 mg.L-1 (2% da CL50-96h). A exposição do jundiá ao NO2 - por 40 dias diminuiu os níveis de lactato no músculo, mas esses níveis aumentaram nos exemplares mantidos em 1,19 mg.L-1 NO2 -. Além disso, os níveis de glicose no músculo e fígado foram significativamente mais baixos nos jundiás expostos à concentração mais elevada de NO2 . Estes resultados indicam que a exposição crônica ao NO2 provoca uma oxidação anaeróbica do substrato para obtenção de energia.(AU)

High nitrite (NO2 -) levels may develop in aquaculture systems due to high fish density, but studies of lethal concentration values and the effect of NO2 - on metabolic parameters and growth are scarce. Consequently, in this study was verified the lethal concentration at 96 h (LC50-96h) for (NO2 -) in juvenile silver catfish, Rhamdia quelen and the effect of four waterborne NO2 - concentrations (0.06, 0.46, 1.19, and 1.52 mg.L-1) on growth, and hepatic and muscular lactate, glucose, glycogen and protein. Nitrite LC50-96h was 20.46 (confidence interval: 16.10-23.68) mg.L-1. In the growth experiment, exposure to NO2 - did not affect weight, length or specific growth rate, but due to mortality (66.7% and 100% after 20 and 40 days, respectively), biomass of juveniles exposed to 1.52 mg.L-1 NO2 - was significantly lower than the biomass of juveniles exposed to other treatments. Therefore, the safe level of nitrite for growth of silver catfish juveniles is below 1.19 mg.L-1 (2% of LC50-96h). Exposure of silver catfish to NO2 - for 40 days reduced lactate levels in muscle, but lactate levels increased in liver tissue of fish maintained at 1.19 mg.L-1 NO2 -. In addition, glucose levels in muscle and liver tissues were significantly lower in silver catfish exposed to the highest NO2 - level. These results indicate that chronic NO2 - exposure causes anaerobic substrate oxidation to meet energy demand.(AU)

Oxidative stress in hypercholesterolemia and its association with Ala16Val superoxide dismutase gene polymorphism.

OBJECTIVES: To investigate the role of the oxidative stress and the antioxidant system as well as the influence of the manganese superoxide dismutase (Ala16Val) polymorphism on hypercholesterolemia. DESIGN AND METHODS: Levels of glucose, lipid, high-sensitivity C reactive protein (hs-CRP), thiobarbituric acid reactive substances (TBARS), carbonyl protein, thiols, reduced glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), and vitamin C, vitamin E, as well as the presence of the manganese superoxide dismutase (Ala16Val) polymorphism were determined in 40 subjects with hypercholesterolemia and 40 controls. RESULTS: Lipid profile, hs-CRP, glucose, TBARS, carbonyl protein, CAT, and vitamin E were significantly higher in subjects with hypercholesterolemia. In contrast, GSH and SOD were lower. TBARS, carbonyl protein, thiols, CAT, and vitamin E were significantly higher in hypercholesterolemic subjects with VV genotype for MnSOD, while GSH, SOD, and vitamin C were lower in these subjects. CONCLUSIONS: We suggest an association between the genotypes of MnSOD, hypercholesterolemia, and oxidative stress biomarkers.

Bioaccumulation and oxidative stress parameters in silver catfish (Rhamdia quelen) exposed to different thorium concentrations.

The objective of this study was to evaluate the effect of chronic thorium (Th) exposure on bioaccumulation, metabolism (through biochemical parameters of the muscle) and oxidative parameters (lipidic peroxidation levels and antioxidant enzymes in the gills and in the hepatic and muscular tissues) of silver catfish (Rhamdia quelen). Silver catfish juveniles were exposed to different waterborne Th levels (in microg L(-1)): 0 (control), 25.3+/-3.2, 80.6+/-12.0, 242.4+/-35.6, and 747.2+/-59.1 for 30 d. The gills and skin were the organs that accumulated the highest Th levels. The increase in the waterborne Th concentration corresponded to a progressive increase in the Th levels in the gills and kidney. Chronic Th exposure causes alterations in the oxidative parameters of silver catfish gills, which are correlated with the Th accumulation in this organ. The levels of GST decreased in the gills of fish exposed to 747.2 microg L(-1) Th and SOD activity decreased in silver catfish exposed to 242.4 and 747.2 microg L(-1) Th. In addition, the increase in the LPO in the gills exposed to 242.4 and 747.2 microg L(-1) Th suggests that higher oxidative damage occurred in the gills. However, in the liver and muscle, these alterations occurred mainly in the lowest waterborne Th level. Metabolic intermediates in the muscle were altered by Th exposure, but no clear relationship was found.

Pesticide contamination of water alters the metabolism of juvenile silver catfish, Rhamdia quelen.

We investigated how pesticide contamination of water affects the metabolism of the silver catfish, Rhamdia quelen, by studying fish maintained at two sites with low and high anthropic activity (Lino Creek, southern Brazil). Several pesticides were found at both stream sites. After 30 days plasma glucose levels were higher in fish exposed to water in the low anthropic activity site than those exposed to water in the high anthropic activity site. Plasma K+ levels, however, were lower after exposure to low anthropic water than after exposure to high anthropic water. Moreover, values of hepatic glycogen, muscle lactate and protein were higher, but glycogen and protein of the kidney were lower in fish exposed to water at the high anthropic activity site. Our results show that these fish can be used as pesticide toxicity indicators in streams near agricultural fields.