Effects of cadmium on glutathione synthesis in hepatopancreas of freshwater crab, Sinopotamon yangtsekiense

Cadmium (Cd) is one of the most deleterious heavy metals in aquatic systems that could promote oxidative damage. To explore the effects of Cd exposure of a freshwater crab (Sinopotamon yangtsekiense) on hepatopancreatic glutathione (GSH) synthesis, crabs were exposed to the reagent with a dose range of 7.25-116.00 mg L(-1) for 48 h. The concentrations of GSH, oxidized glutathione (GSSG), NADPH and NADP(+), as well as the activities of enzymes involved in GSH synthesis, i.e. glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G6PD), gamma-glutamylcysteine synthetase (gamma-GCS) were determined. Progressive depletion of cellular GSH content was observed with the increasing of Cd concentrations, while the level of GSSG remained constant. In response to Cd exposure, crabs showed significant induction of G6PD and NADPH, however, only up to moderate exposures. GR activity remained at a steady level at all exposure concentrations. The activity of gamma-GCS was significantly positively correlated with the Cd concentration. These results suggested that GSH synthesis could be activated against reactive oxygen species induced by lower Cd exposure; under the higher Cd exposure conditions, an inhibition of NADPH-dependant redox cycling and de novo GSH synthesis led to significant decrease in GSH content.     

Glutathione and its dependent enzymes’ modulatory responses to toxic metals and metalloids in fish—a review

Toxic metals and metalloid are being rapidly added from multiple pathways to aquatic ecosystem and causing severe threats to inhabiting fauna including fish. Being common in all the type of aquatic ecosystems such as freshwater, marine and brackish water fish are the first to get prone to toxic metals and metalloids. In addition to a number of physiological/biochemical alterations, toxic metals and metalloids cause enhanced generation of varied reactive oxygen species (ROS) ultimately leading to a situation called oxidative stress. However, as an important component of antioxidant defence system in fish, the tripeptide glutathione (GSH) directly or indirectly regulates the scavenging of ROS and their reaction products. Additionally, several other GSH-associated enzymes such as GSH reductase (GR, EC 1.6.4.2), GSH peroxidase (EC 1.11.1.9), and GSH sulfotransferase (glutathione-S-transferase (GST), EC 2.5.1.18) cumulatively protect fish against ROS and their reaction products accrued anomalies under toxic metals and metalloids stress conditions. The current review highlights recent research findings on the modulation of GSH, its redox couple (reduced glutathione/oxidised glutathione), and other GSH-related enzymes (GR, glutathione peroxidase, GST) involved in the detoxification of harmful ROS and their reaction products in toxic metals and metalloids-exposed fish.     

Impaired glutathione redox status is associated with decreased survival in two organophosphate-poisoned marine bivalves

Biomonitoring organophosphate (OP) exposure in marine environments is generally achieved by the measurement of acetylcholinesterase activity in bivalves like mussels. However, there is evidence that indicates that oxidative stress may be implied in OP toxicity. The aim of this study was to evaluate the relationship between survival from the OP insecticide fenitrothion and glutathione levels in marine bivalves. Mussels (Mytilus galloprovincialis Lam.) and scallops (Flexopecten flexuosus Poli) were exposed, in a time to death test, to their LC85 of fenitrothion for 96 h. OP-poisoned mussels showed reduced (GSH) and oxidised (GSSG) glutathione depletion in the digestive gland, muscle and gills. Pectinid spats exposed to this insecticide presented GSH depletion in the digestive gland and mantle, and a reduction of the GSH/GSSG ratio in gills and mantle. Although survival curves were significantly different and mussels withstood twice as much fenitrothion as pectinid spats, muscular GSH/GSSG ratio was highly related to mortality in both species. We suggest that an impairment in the glutathione redox status could result in an induction of the cell death, either by apoptosis or necrosis, leading ultimately to the death of the organism. We conclude that whereas glutathione depletion can be used as a biomarker of exposure, the muscular GSH/GSSG ratio might be used as a biochemical marker of effect and individual susceptibility to mortality of marine bivalves exposed to fenitrothion or other pollutants that induce oxidative stress.     

Oxidative stress and detoxification biomarker responses in aquatic freshwater vertebrates exposed to microcystins and cyanobacterial biomass

Cyanobacterial blooms represent a serious threat to the aquatic environment. Among other effects, biochemical markers have been studied in aquatic vertebrates after exposures to toxic cyanobacteria. Some parameters such as protein phosphatases may serve as selective markers of exposure to microcystins, but under natural conditions, fish are exposed to complex mixtures, which affect the overall biomarker response. This review aims to provide a critical summary of biomarker responses in aquatic vertebrates (mostly fish) to toxic cyanobacteria with a special focus on detoxification and oxidative stress. Detoxification biomarkers such as glutathione (GSH) and glutathione-S-transferase (GST) showed very high variability with poor general trends. Often, stimulations and/or inhibitions and/or no effects at GSH or GST have been reported, even within a single study, depending on many variables, including time, dose, tissue, species, etc. Most of the oxidative stress biomarkers (e.g., superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) provided more consistent responses, but only lipid peroxidation (LPO) seemed to fulfill the criteria needed for biomarkers, i.e., a sufficiently long half-life and systematic response. Indeed, reviewed papers demonstrated that toxic cyanobacteria systematically elevate levels of LPO, which indicates the important role of oxidative damage in cyanobacterial toxicity. In summary, the measurement of biochemical changes under laboratory conditions may provide information on the mode of toxic action. However, comparison of different studies is very difficult, and the practical use of detoxification or oxidative stress biomarkers as diagnostic tools or early warnings of cyanobacterial toxicity is questionable.     

Hydroxyl radical generation and oxidative stress in Carassius auratus liver as affected by 2,4,6-trichlorophenol

With Carassius auratus, one of the main economic fish species in Eastern China as test material, this paper studied the hydroxyl radical generation and oxidative stress in its liver under the effect of 2,4,6-trichlorophenol (2,4,6-TCP). Different doses of 2,4,6-TCP were injected intraperitoneally into the fishes, and the Electron paramagnetic resonance (EPR) spectra of hepatic free radicals, activities of superoxide dismutase (SOD), catalase (CAT) and glutathione-s-transferase (GST), levels of reduced glutathione (GSH) and oxidized glutathione (GSSG), and malondialdehyde (MDA) contents were determined 24h after injection. The results showed that under the effects of 2,4,6-TCP, the generation of free radical that was considered to be hydroxyl radical increased significantly, the activities of antioxidant enzymes decreased, with CAT most strongly affected and followed by SOD and GST, the GSH level decreased significantly while GSSG level had little difference, resulting in a decreased GSH/GSSG ratio, and the MDA content increased significantly. All the test parameters showed that C. auratus was subjected to oxidative stress and damage when exposed to 2,4,6-TCP.     

Effects of hexachlorobenzene on antioxidant status of liver and brain of common carp (Cyprinus carpio)

Hexachlorobenzene (HCB)-induced oxidative damages have been published in rats while the effects have not yet been reported in fishes. Juvenile common carps (Cyprinus carpio) were exposed to waterborne HCB from 2 to 200 microg l-1 for 5, 10 or 20 days. Liver and brain were analyzed for various parameters of oxidative stress. There were no significant changes of glutathione (GSH) content and superoxide dismutase (SOD) activity in liver after 5 or 10 days exposure, whereas obvious drops were observed at higher concentrations after 20 days exposure. Significant decreases of GSH content and SOD activity in brain were found during all the exposure days. In brain, HCB also significantly elevated the contents of reactive oxygen species (ROS), thiobarbituric acid- reactive substances (TBARS, as an indicator of lipid peroxidation products), glutathione disulfide (GSSG), and activities of nitric oxide synthase (NOS), glutathione peroxidase (GPx), and glutathione reductase (GR), and inhibited activities of acetylcholinesterase (AchE) and glutathione S-transferase (GST). The results clearly demonstrated that environmentally possible level of HCB could result in oxidative stress in fish and brain was a sensitive target organ of HCB toxicity.     

Effects of methylmercury exposure on glutathione metabolism, oxidative stress, and chromosomal damage in captive-reared common loon (Gavia immer) chicks

We quantified the level of dietary mercury (Hg), delivered as methylmercury chloride (CH₃HgCl), associated with negative effects on organ and plasma biochemistries related to glutathione (GSH) metabolism and oxidative stress, and chromosomal damage in captive-reared common loon (Gavia immer) chicks reared from hatch to 105 days. Mercury-associated effects related to oxidative stress and altered glutathione metabolism occurred at 1.2 μg Hg/g and 0.4 μg Hg/g, an ecologically relevant dietary mercury level, but not at 0.08 μg Hg/g. Among the variables that contributed most to dissimilarities in tissue chemistries between control and treatment groups were increased levels of oxidized glutathione (GSSG), GSH peroxidase, and the ratio of GSSG to GSH in brain tissue; increased levels of hepatic GSH; and decreased levels of hepatic glucose-6-phosphate dehydrogenase (G-6-PDH). Our results also suggest that chronic exposure to environmentally relevant dietary Hg levels did not result in statistically significant somatic chromosomal damage in common loon chicks.     

Proteomic responses to lead-induced oxidative stress in Talinum triangulare Jacq. (Willd.) roots: identification of key biomarkers related to glutathione metabolisms

In this study, Talinum triangulare Jacq. (Willd.) treated with different lead (Pb) concentrations for 7 days has been investigated to understand the mechanisms of ascorbate–glutathione metabolisms in response to Pb-induced oxidative stress. Proteomic study was performed for control and 1.25 mM Pb-treated plants to examine the root protein dynamics in the presence of Pb. Results of our analysis showed that Pb treatment caused a decrease in non-protein thiols, reduced glutathione (GSH), total ascorbate, total glutathione, GSH/oxidized glutathione (GSSG) ratio, and activities of glutathione reductase and γ-glutamylcysteine synthetase. Conversely, cysteine and GSSG contents and glutathione-S-transferase activity was increased after Pb treatment. Fourier transform infrared spectroscopy confirmed our metabolic and proteomic studies and showed that amino, phenolic, and carboxylic acids as well as alcoholic, amide, and ester-containing biomolecules had key roles in detoxification of Pb/Pb-induced toxic metabolites. Proteomic analysis revealed an increase in relative abundance of 20 major proteins and 3 new proteins (appeared only in 1.25 mM Pb). Abundant proteins during 1.25 mM Pb stress conditions have given a very clear indication about their involvement in root architecture, energy metabolism, reactive oxygen species (ROS) detoxification, cell signaling, primary and secondary metabolisms, and molecular transport systems. Relative accumulation patterns of both common and newly identified proteins are highly correlated with our other morphological, physiological, and biochemical parameters.     

Effects of malathion and chlorpyrifos on acetylcholinesterase and antioxidant defense system in Oxya chinensis (Thunberg) (Orthoptera: Acrididae)

The study was undertaken to evaluate the effects of malathion and chlorpyrifos on acetylcholinesterase (AChE), esterase (EST) activity and antioxidant system after topical application with different concentration to Oxya chinensis. The results showed that malathion and chlorpyrifos inhibited EST, AChE activity and increased malondialdehyde (MDA) contents. A change in superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), glutathione peroxidase (GPx), and glutathione reductase (GR) activity combined with reduced glutathione (GSH) and total glutathione (tGSH) contents was found in O. chinensis after malathion and chlorpyrifos treatments. Malathion and chlorpyrifos increased SOD and CAT activity compared with the control. With the concentrations increasing, SOD and CAT activity showed the similar tendency, namely, SOD and CAT activity increased at the lower concentrations and decreased at the higher concentrations. The results showed that malathion and chlorpyrifos decreased significantly GR activity. GST and GPx activity at the studied concentrations of chlorpyrifos was lower than that of the control. However, no significance was observed. GPx and GST activity in malathion treated grasshoppers showed a biphasic response with an initial increase followed by a decline in its activity. Malathion and chlorpyrifos decreased GSH contents and the ratio of GSH/GSSG. The present findings indicated that the toxicity of malathion and chlorpyrifos might be associated with oxidative stress.     

Glutathione levels and enzyme activity in the tissues of bank vole Clethrionomys glareolus chronically exposed to a mixture of metal contaminants

The biochemical response to chronic heavy metal exposure was studied in tissues of bank voles Clethrionomys glareolus. Animals were collected from three sites located 4, 8 and 30km from a zinc-lead smelter, the area's main source of metal contamination. Concentrations of Cd, Pb, Zn and Fe were measured in the liver, kidneys and gonads to assess the level of metal intoxication. In response to intoxication, organisms activate detoxification mechanisms which can protect animals from metals' toxicity. Glutathione plays an important role in toxic substance detoxification. Total glutathione (tGSH) and glutathione disulfide (GSSG) were measured in the tissues. Also, the activity of glutathione reductase (GR), glutathione peroxidase (GPX), and glutathione-S-transferase (GST) was measured in the studied tissues. Results indicate that levels of all studied parameters were tissue and site-dependent. Evidence indicates that the most sensitive parameter of metal toxicity for animals living in a chronically contaminated environment is the GSH/GSSG ratio. In our study, the GSH/GSSG ratio was decreased in the liver of animals with high Cd levels. However, the relationship between Pb and the GSH/GSSG ratio was positive in the gonads. Cadmium and lead negatively influenced GPX activity in the liver; this was probably connected with inhibition of the Se-dependent fraction. The relationship between iron and GR activity in the kidney was also negative, but other correlations for iron both in liver and kidney were not significant. Positive correlations between Zn levels and GST and GR activity were found in the gonads of bank voles.     

Glutathione status and glutathione reductase activity in spruce needles of healthy and damaged trees at two mountain sites

Levels of glutathione, in both reduced and oxidized form, and glutathione reductase activity were monitored in needles of healthy and damaged spruce trees (Picea abies (L.) Karst.) during the course of four vegetation periods at two natural sites. The glutathione content and glutathione reductase activity showed a pronounced annual rhythm in undamaged trees, whereas damaged spruce trees deviated significantly from this course. In comparison with undamaged trees, damaged trees showed markedly increased levels of glutathione during the test period of 1989-1991. However, glutathione reductase activity differed in damaged and undamaged trees, only in 1989-1990. The ratio of reduced to oxidized glutathione (GSH/GSSG ratio) was slightly higher in damaged trees, and the highest levels were found during the winter months. In the case of damaged trees, a correlation between GSH/GSSG ratio and current ozone levels at the sites could be clearly established. The present results indicate that damaged trees suffer from increased oxidative stress, especially in the period from June to October.     

Effects of chronic exposure of 2,4-dichlorophenol on the antioxidant system in liver of freshwater fish Carassius auratus

There were few reports on the antioxidant response of aquatic organisms exposed to 2,4-dichlorophenol (2,4-DCP). This research explored the hepatic antioxidant responses of fish to long-term exposure of 2,4-DCP for the first time. Freshwater fish Carassius auratus were chosen as experimental animals. The fish were exposed to six different concentrations of 2,4-DCP (0.005-1.0 mg/l) for 40 days and then liver tissues were separated for determination. As shown from the results, 40 days afterwards, the activities of catalase (CAT) and selenium-dependent glutathione peroxidase (Se-GPx) and the content of oxidized glutathione (GSSG) were induced significantly on the whole compared to control group; superoxide dismutase (SOD) responded to 2,4-DCP exposure at only 0.005 mg/l; the content of reduced glutathione (GSH) was suppressed continuously except Group 7; the activity of glutathione reductase was inhibited initially and then restored to control level from Group 4 on; glutathione S-transferase had only slight responses in Groups 3 and 4. Total glutathione (tGSH) and GSH/GSSG ratio were also calculated to analyze the occurrence of oxidative stress. Besides, good dose-effect relations, which cover most of the exposure concentration range, were found between 2,4-DCP level and CAT activity, GSSG content, Se-GPx activity, respectively. In conclusion, SOD and Se-GPx may be potential early biomarkers of 2,4-DCP contamination in aquatic ecosystems, and further studies will be necessary.     

Glutathione-dependent resistance of the European eel Anguilla anguilla to the herbicide molinate.

Eels of species Anguilla anguilla were exposed to 5/4 LC50 (41.8 mg/l) of the herbicide molinate for 96 h in a time to death (TTD) test. Glutathione content (GSx, GSH, GSSG), glutathione reductase (GR) and gamma-glutamyl transpeptidase (gamma-GT) activities were determined in the liver and muscle tissues of dead and surviving (intoxicated) animals and compared to control values (non-exposed eels). TTD was positively correlated to hepatic GSH, GSH:GSSG ratio, hepatic and muscular GR, but negatively correlated to muscular GSH, which was severely depleted. Furthermore, glutathione and enzyme activities were intercorrelated, especially GSH and GR. These results indicate that eels which were able to induce GR activity, increase GSH and maintain the GSH:GSSG ratio in the liver showed an extended survival under the oxidative stress generated by molinate than those that lost glutathione homeostasis.     

Lead toxicity, defense strategies and associated indicative biomarkers in Talinum triangulare grown hydroponically

Talinum species have been used to investigate a variety of environmental problems for e.g. determination of metal pollution index and total petroleum hydrocarbons in roadside soils, stabilization and reclamation of heavy metals (HMs) in dump sites, removal of HMs from storm water-runoff and green roof leachates. Species of Talinum are popular leaf vegetables having nutrient antinutrient properties. In this study, Talinum triangulare (Jacq.) Willd (Ceylon spinach) grown hydroponically were exposed to different concentrations of lead (Pb) (0, 0.25, 0.5, 0.75, 1.0 and 1.25 mM) to investigate the biomarkers of toxicity and tolerance mechanisms. Relative water content, cell death, photosynthetic pigments, sulphoquinovosyldiacylglycerol (SQDG), anthocyanins, α-tocopherol, malondialdehyde (MDA), reactive oxygen species (ROS) glutathione (GSH and GSSG) and elemental analysis have been investigated. The results showed that Pb in roots and shoots gradually increased as the function of Pb exposure; however Pb concentration in leaves was below detectable level. Chlorophylls and SQDG contents increased at 0.25 mM of Pb treatment in comparison to control at all treated durations, thereafter decreased. Levels of carotenoid, anthocyanins, α-tocopherol, and lipid peroxidation increased in Pb treated plants compared to control. Water content, cells death and elemental analysis suggested the damage of transport system interfering with nutrient transport causing cell death. The present study also explained that Pb imposed indirect oxidative stress in leaves is characterized by decreases in GSH/GSSG ratio with increased doses of Pb treatment. Lead-induced oxidative stress was alleviated by carotenoids, anthocyanins, α-tocopherol and glutathione suggesting that these defense responses as potential biomarkers for detecting Pb toxicity.     

Effects of heavy metals and nitroaromatic compounds on horseradish glutathione S-transferase and peroxidase

Glutathione S-transferase (GST) and peroxidase (POX) activities have a direct relation to the effect of stress on plant metabolism. Changes in the activities of the enzymes were therefore studied. Horseradish hairy roots were treated by selected bivalent ions of heavy metals (HMs) and nitroaromatic compounds (NACs). We have shown differences in GST activity when assayed with substrates 1-chloro-2,4-dinitrobenzene (CDNB) and 1,2-dichloro-4-nitrobenzene (DCNB). The conjugation of DCNB catalysed by GST was inhibited in all roots treated with HMs as compared to non-treated roots, whereas NACs caused induction of the activity in dependence on the exposition time and concentration of compounds. The conjugation of CDNB by GST was not affected to the same extent. The increase of GST activity was determined in cultures treated by nickel (0.1 mM) and diaminonitrotoluenes (DANTs, 0.1 mM) for 6 h, whereas the roots treated by 2,4,6-trinitrotoluene (TNT), 4-amino-2,6-dinitrotoluene (ADNT) and dinitrotoluene (DNT, 1.0 mM) needed 27 h treatment to induce the activity. The POX activity of cultures treated by HMs was inhibited to 17-35% in comparison to non-treated cultures. The POX activity of roots treated by TNT (0.1 and 1.0 mM) for 6 and 27 h and by ADNT (0.1 and 1.0 mM) for 6 h was inhibited. A partial increase of POX activity was measured in roots treated by all NACs for 27 h. The content of oxidized glutathione (GSSG) and reduced glutathione (GSH) in the roots differed significantly. It was followed as a symptom of the stress reaction of the plant metabolism to the effect of NACs and HMs.     

Anguilla anguilla L. oxidative stress biomarkers responses to copper exposure with or without beta-naphthoflavone pre-exposure

The redox cycling of heavy metals as well as their interactions with organic pollutants is a major contributor to the oxidative stress resulting from aquatic pollution. Therefore, in order to evaluate beta-naphthoflavone (BNF), Cu and BNF/Cu-induced oxidative stress with single and subsequent exposures, research was carried out in European eel (Anguilla anguilla L.). Eel gill and kidney oxidative stress biomarker responses such as lipid peroxidation (LPO), glutathione peroxidase (GPX), catalase (CAT), glutathione S-transferase (GST) and total reduced glutathione (GSH) to a single 24 h exposure to two copper concentrations (Cu-1 microM, 2.5 microM) and BNF (2.7 microM) with or without 24 h BNF (2.7 microM) pre-exposure were investigated. Cu exposure alone showed a significant gill GST increase at the lowest concentration and GSH content decrease for the highest concentration. Double BNF exposure in gill demonstrated a significant increase in LPO, CAT, GPX and GST, as well as a decrease in GSH content. However, in sequential BNF/Cu exposures, only the highest Cu concentration exhibited a significant increase in LPO and GSH as well as a decrease in GPX (vs. BNF + CW). In kidney, Cu exposure alone showed a significant CAT and GSH contents decrease for both concentrations, and at highest concentration in GPX; as well as GST increase at the lowest concentration. Double BNF exposure showed a significant increase in LPO and GST. Nevertheless, in sequential BNF/Cu exposures, both concentrations exhibited a significant increase in LPO and decrease in GSH contents. Moreover, LPO was also increased significantly in comparison to BNF+CW and the equivalent Cu exposures without BNF pre-exposure. Concerning GPX, a significant increase was observed at highest Cu concentration. In GST, a significant decrease at the lowest Cu concentration and increase at the highest Cu concentration was observed. Summarizing, a simple copper or BNF exposures have no ability to induce LPO in both gill and kidney. However, double BNF exposure induced LPO in both organs and sequential BNF/Cu exposures potentiated the risk of peroxidative damage occurrence in both organs. BNF/Cu interference on antioxidant responses differs between the studied organs. In gill, antagonistic effects were denoted with probable reflex in terms of peroxidative damage increase. In kidney, BNF pre-exposure prevented CAT and GPX inhibition by copper; though, no advantage of this effect was perceptible as defence against LPO generation. Considering BNF as a surrogate for a PAH and the detected interactions with copper, as well as the likelihood that these effects would be observed in polluted ecosystems, current results demonstrate their relevance to actual ecological exposures contributing to a better knowledge on oxidative stress mechanisms in fish.     

Oxidative stress, liver biotransformation and genotoxic effects induced by copper in Anguilla anguilla L.--the influence of pre-exposure to beta-naphthoflavone

Fish are exposed in the aquatic ecosystems to different classes of pollutants. Polycyclic aromatic hydrocarbons (PAHs) and heavy metals represent two important classes of aquatic contaminants. Thus, one lot of European eels (Anguilla anguilla L.) was pre-exposed during 24 h to 2.7 microM beta-naphthoflavone (BNF; a PAH-like compound), and subsequently exposed during 24 h to 0, 1 and 2.5 microM copper (Cu). Additionally, another lot not pre-exposed to BNF was exposed to the same Cu concentrations. BNF pre-exposure promoted a significant increase in liver ethoxyresorufin O-deethylase (EROD) activity, but did not change the other responses investigated in eels. On the other hand, both Cu concentrations did not modify the liver EROD activity either in eels pre-exposed to BNF or not. Liver total cytochrome P450 was increased in eels exposed to Cu 2.5 microM, being significantly only in eels not pre-exposed to BNF. Free sulfhydryl group content was decreased by 1 and 2.5 microM in eels pre-exposed to BNF or not pre-exposed, being significant at 2.5 microM Cu in eels not pre-exposed compared to its control. Liver total glutathione (TG), reduced glutathione (GSH) and GSH/oxidized glutathione (GSSG) levels were slightly decreased by 1 and 2.5 microM Cu in eels pre-exposed to BNF, whereas a slight tendency to increase was observed in eels not pre-exposed. Thus, liver TG and GSH significantly decreased in 2.5 microM Cu BNF pre-exposed eels compared to eels not pre-exposed to BNF. Liver glutathione reductase and catalase activities were significantly inhibited by 1 and 2.5 microM Cu in eels pre-exposed to BNF, concomitantly with a slight liver glutathione peroxidase tendency to decrease. Lipid peroxidation was significantly increased by 1 microM Cu in eels either pre-exposed or not pre-exposed to BNF. Liver H(2)O(2) was significantly increased by 1 microM Cu in eels pre-exposed to BNF. Liver DNA integrity was significantly decreased by 1 and 2.5 microM Cu in eels pre-exposed to BNF. The oxidative stress and genotoxic effects induced by Cu in eels pre-exposed to BNF revealed that the metal effects are potentiated by previous exposure to BNF.     

2-chlorophenol induced ROS generation in fish Carassius auratus based on the EPR method

In the present study, a secondary spin trapping technique was used followed by electron paramagnetic resonance (EPR) analysis, to study the potential of reactive oxygen species (ROS) production after fish (Carassius auratus) were injected i.p. with different doses (50, 100, 200, 250, 500mgkg(-1)) of 2-chlorophenol (2-CP). The ROS signal intensity of the EPR spectrum showed a significant increase (p<0.05, compared with the control) when the 2-CP dose was as low as 50mgkg(-1). There is a good relationship between the 2-CP administered doses and ROS generation. Based on the hyperfine splitting constants and shape of the EPR spectrum, the ROS which was generated in fish liver after intraperitoneal (i.p.) injection of 2-CP was identified as ()OH. SOD and CAT activities were found to be induced at lower doses of 2-CP. GSH levels fell below the control level following all treatments with 2-CP, and GSSG levels changed along with those of GSH. These observations indicated that the fish experienced oxidative stress. The strong positive correlation (r=0.966, p<0.005) between ()OH radical and lipid peroxidation suggested that lipid peroxidation was possibly induced by ()OH. The phase II detoxification enzyme glutathione-S-transferase (GST) may play an important role in 2-CP metabolism or excretion and, consequently, reduce ROS production. This study provides strong evidence that level of ROS is significantly increased in 2-CP stressed fish, and ROS may serve as a potential biomarker to indicate 2-CP contamination.     

Cytotoxic effect of fenitrothion and lambda-cyhalothrin mixture on lipid peroxidation and antioxidant defense system in rat kidney

A mixture of pyrethroids plus organophosphates was assessed for their potential effects on lipid peroxidation, the antioxidant defense system and lactate dehydrogenase (LDH) in rat kidney in vitro. Various insecticide concentrations were incubated with kidney homogenate at 37°C for different incubation times. Treatment with fenitothion (FNT) plus lambda-cyhalothrin (LC) caused a significant induction (P < 0.05) in thiobarbituric acid reactive substances (TBARS), which might be associated to decreased levels of reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST) activities and protein content in rat kidney. However, a significant induction of lactate dehydrogenase (LDH) activity was observed. The effect was concentration and time dependent. It can be concluded that depletion of GSH might indicate that reactive oxygen species (ROS) could be involved in the toxic effects of FNT plus LC which lead to marked perturbations in antioxidant defense system.