Relating diving behavior and antioxidant status: Insights from oxidative stress biomarkers in the blood of two distinct divers, Mirounga leonina and Arctocephalus australis.

Pinnipeds rely upon diving to perform essential activities, including foraging. As pulmonated animals, oxygen privation experienced during submergence represents a considerable challenge both physiologically and biochemically. Routine exposure to hypoxia and the rapid transitions between ischemia/reperfusion of tissues leads to extremely high reactive oxygen species (ROS) production, which must be opposed by antioxidant defenses to avoid oxidative stress. The diving behaviors and capabilities of pinnipeds are very diverse, resulting in distinct metabolic responses among species. To assess whether these characteristics reflect the antioxidant status of two marine diving mammals with distinct diving capacities, oxidative stress biomarkers were measured in the blood of Arctocephalus australis (n=11) and Mirounga leonina (n=12). All of the biomarkers analyzed in M. leonina were significantly higher than those of A. australis, suggesting that higher antioxidant content is needed to counteract the high ROS production associated with the long submergence times (24.3±5.6min) of this species, which are nearly ten times greater than those of A. australis (2.8±0.5min). Thus, the constitutive antioxidant defenses of both species are of distinct magnitudes due to their inherent diving capacity.

Antioxidant therapy attenuates oxidative stress in the blood of subjects exposed to occupational airborne contamination from coal mining extraction and incineration of hospital residues.

Coal mining and incineration of solid residues of health services (SRHS) generate several contaminants that are delivered into the environment, such as heavy metals and dioxins. These xenobiotics can lead to oxidative stress overgeneration in organisms and cause different kinds of pathologies, including cancer. In the present study the concentrations of heavy metals such as lead, copper, iron, manganese and zinc in the urine, as well as several enzymatic and non-enzymatic biomarkers of oxidative stress in the blood (contents of lipoperoxidation = TBARS, protein carbonyls = PC, protein thiols = PT, α-tocopherol = AT, reduced glutathione = GSH, and the activities of glutathione S-transferase = GST, glutathione reductase = GR, glutathione peroxidase = GPx, catalase = CAT and superoxide dismutase = SOD), in the blood of six different groups (n = 20 each) of subjects exposed to airborne contamination related to coal mining as well as incineration of solid residues of health services (SRHS) after vitamin E (800 mg/day) and vitamin C (500 mg/day) supplementation during 6 months, which were compared to the situation before the antioxidant intervention (Ávila et al., Ecotoxicology 18:1150-1157, 2009; Possamai et al., Ecotoxicology 18:1158-1164, 2009). Except for the decreased manganese contents, heavy metal concentrations were elevated in all groups exposed to both sources of airborne contamination when compared to controls. TBARS and PC concentrations, which were elevated before the antioxidant intervention decreased after the antioxidant supplementation. Similarly, the contents of PC, AT and GSH, which were decreased before the antioxidant intervention, reached values near those found in controls, GPx activity was reestablished in underground miners, and SOD, CAT and GST activities were reestablished in all groups. The results showed that the oxidative stress condition detected previously to the antioxidant supplementation in both directly and indirectly subjects exposed to the airborne contamination from coal dusts and SRHS incineration, was attenuated after the antioxidant intervention.

Occupational airborne contamination in south Brazil: 1. Oxidative stress detected in the blood of coal miners.

Reactive oxygen species and nitrogen species have been implicated in the pathogenesis of coal dust-induced toxicity. The present study investigated several oxidative stress biomarkers (Contents of lipoperoxidation = TBARS, reduced = GSH, oxidized = GSSG and total glutathione = TG, alpha-tocopherol, and the activities of glutathione S-transferase = GST, glutathione reductase = GR, glutathione peroxidase = GPx, catalase = CAT and superoxide dismutase = SOD), in the blood of three different groups (n = 20 each) exposed to airborne contamination associated with coal mining activities: underground workers directly exposed, surface workers indirectly exposed, residents indirectly exposed (subjects living near the mines), and controls (non-exposed subjects). Plasma TBARS were increased and whole blood TG and GSH levels were decreased in all groups compared to controls. Plasma alpha-tocopherol contents showed approximately half the values in underground workers compared to controls. GST activity was induced in workers and also in residents at the vicinity of the mining plant, whilst CAT activity was induced only in mine workers. SOD activity was decreased in all groups examined, while GPx activity showed decreased values only in underground miners, and GR did not show any differences among the groups. The results showed that subjects directly and indirectly exposed to coal dusts face an oxidative stress condition. They also indicate that people living in the vicinity of the mine plant are in health risk regarding coal mining-related diseases.

Occupational airborne contamination in South Brazil: 2. Oxidative stress detected in the blood of workers of incineration of hospital residues.

One of the most useful methods for elimination of solid residues of health services (SRHS) is incineration. However, it also provokes the emission of several hazardous air pollutants such as heavy metals, furans and dioxins, which produce reactive oxygen species and oxidative stress. The present study, which is parallel to an accompanied paper (Avila Jr. et al., this issue), investigated several enzymatic and non-enzymatic biomarkers of oxidative stress in the blood (contents of vitamin E, lipoperoxidation = TBARS, reduced glutathione = GSH, oxidized glutathione = GSSG, and activities of glutathione S-transferase = GST, glutathione reductase = GR, glutathione peroxidase = GPx, catalase = CAT and superoxide dismutase = SOD), in three different groups (n = 20 each) exposed to airborne contamination associated with incineration of SRHS: workers directly (ca. 100 m from the incinerator) and indirectly exposed (residents living ca. 5 km the incineration site), and controls (non-exposed subjects). TBARS and GSSG levels were increased whilst GSH, TG and alpha-tocopherol contents were decreased in workers and residents compared to controls. Increased GST and CAT activities and decreased GPx activities were detected in exposed subjects compared to controls, while GR did not show any difference among the groups. In conclusion, subjects directly or indirectly exposed to SRHS are facing an oxidative insult and health risk regarding fly ashes contamination from SRHS incineration.

Oxidative stress after acute and sub-chronic malathion intoxication in Wistar rats.

Malathion is an insecticide of the group of organophosphate pesticides (OPs), which shows strong insecticidal effects. However, it possesses mutagenic and carcinogenic properties and shows organ-specific toxicity in relation to the heart, kidney and other vertebrate organs. The exact mechanism of the genotoxic effects of malathion is not yet known. Free radical damage is an important direct or indirect factor in several pathological and toxicological processes, including malathion poisoning. The aim of the present study was the evaluation of oxidative damage in different tissues of Wistar rats, administered intra peritoneally at doses of 25, 50, 100 and 150mgmalathion/kg, after acute and sub-chronic malathion exposure. Oxidative stress evaluation was based on lipid peroxidation by levels of thiobarbituric acid reactive substances (TBARS), protein oxidation by levels of carbonyl groups, and also on the activities of superoxide dismutase and catalase, two antioxidant enzymes that detoxity superoxide radical (O(2)(-)) and hydrogen peroxide, respectively. The results showed that the most sensitive targets of oxidative damage were kidney, lung and diaphragm after acute treatment, and liver, quadriceps and serum after sub-chronic treatment. Also, in general, increased lipid peroxidation measured as TBARS levels seems to be a better biomarker of oxidative stress compared to the contents of protein carbonyls after acute and sub-chronic malathion treatments. The present findings reinforce the concept that oxidative stress and particularly lipoperoxidation, are involved in OPs toxicity.

Albendazole as a promising molecule for tumor control.

This work evaluated the antitumor effects of albendazole (ABZ) and its relationship with modulation of oxidative stress and induction of DNA damage. The present results showed that ABZ causes oxidative cleavage on calf-thymus DNA suggesting that this compound can break DNA. ABZ treatment decreased MCF-7 cell viability (EC50=44.9 for 24h) and inhibited MCF-7 colony formation (~67.5% at 5µM). Intracellular ROS levels increased with ABZ treatment (~123%). The antioxidant NAC is able to revert the cytotoxic effects, ROS generation and loss of mitochondrial membrane potential of MCF-7 cells treated with ABZ. Ehrlich carcinoma growth was inhibited (~32%) and survival time was elongated (~50%) in animals treated with ABZ. Oxidative biomarkers (TBARS and protein carbonyl levels) and activity of antioxidant enzymes (CAT, SOD and GR) increased, and reduced glutathione (GSH) was depleted in animals treated with ABZ, indicating an oxidative stress condition, leading to a DNA damage causing phosphorylation of histone H2A variant, H2AX, and triggering apoptosis signaling, which was confirmed by increasing Bax/Bcl-xL rate, p53 and Bax expression. We propose that ABZ induces oxidative stress promoting DNA fragmentation and triggering apoptosis and inducing cell death, making this drug a promising leader molecule for development of new antitumor drugs.

Time-dependent oxidative stress caused by benznidazole.

Benznidazole (BZN) is a nitroimidazole derivative which has a notable trypanocide activity, and it is the only drug used in Brazil and Argentina for the treatment of Chagas' disease. The drug in current use is thought to act, at least in part, by inducing oxidative stress within the parasite. Imidazolic compounds are involved in the production of reactive oxygen species (ROS). In order to evaluate the effect of BZN on ROS production and on the antioxidant status of the host, male rats were treated for different periods of time (2, 4, 6, 10 and 30 days) with 40 mg BZN/kg body weight. After treatment, biomarkers of oxidative stress such as the activities of catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) and glutathione reductase (GR), and also thiobarbituric acid reactive species (TBARS), reduced glutathione (GSH), total glutathione (TG) and oxidized glutathione (GSSG) concentrations, were measured in crude hepatic homogenates. Our results revealed that BZN is able to cause tissue damage as shown by increased TBARS content, inhibition of some antioxidants and induction of other antioxidants in a concentration- and time-dependent manner. The tissue damage measured as TBARS increased up to the 10th day of treatment. GST activity was inhibited during the BZN treatment. On the other hand, CAT and GR showed similar increased activities at the beginning, followed by decreased activities at the end of the treatment. After 30 days of treatment, GR activity remained low while CAT activity was high, compared to controls. The SOD activities remained unchanged throughout the experimental period. GSH showed lower values at the beginning of BZN treatment but the hepatic concentrations were enhanced at the end of the experimental period. Total glutathione showed a similar profile, and oxidized glutathione showed higher values in rats treated with BZN. In conclusion, these results indicate that, at therapeutic doses, BZN treatment elicits an oxidative stress in rat hepatocytes.

Complexes trans-[RuCl(2)(nic)(4)] and trans-[RuCl(2)(i-nic)(4)] as free radical scavengers.

This study evaluates the action of the new ruthenium complexes trans-RuCl(2)(nic)(4)] (I) and trans-[RuCl(2)(i-nic)(4)] (II) as free radical scavengers. In our experiments, both compounds acted as scavengers of superoxide anion (O(2)*(-)), hydroxyl radicals (HO*) and nitrogen monoxide (formally known as 'nitric oxide'; NO*). In addition, complexes I and II potentiated the release of NO* from S-nitroso-N-acetyl-DL-penicilamine (SNAP), a NO* donor. Complex II, but not I, also decreased the nitrite levels in culture media of activated macrophages. A hypsochromic shift of lambda(max) and a significant change in half-wave potential (E(1/2)) was observed when NO* was added to the Complex II. Thiobarbituric reactive substance (TBARS) levels were significantly reduced in rats treated for 1 week with Complex II plus tert-butylhydroperoxide, when compared to rats treated only with tert-butylhydroperoxide. None of the complexes showed cytotoxicity. These findings support the suggestion that the new ruthenium complexes, especially trans-[RuCl(2)(i-nic)(4)] or its derivatives, might provide potential therapeutic benefits in disorders where reactive nitrogen (RNS) or oxygen (ROS) species are involved.

Fish antioxidant defenses--a comparative approach.

There are few comparative studies of vertebrate antioxidant defenses (AD) in the literature. Enzymatic (superoxide dismutase, SOD, and catalase, CAT) and non-enzymatic (alpha-tocopherol, beta-carotene, ubiquinol 10 and blood glutathione) antioxidant defenses were investigated in the liver and blood of 37 fish species, 15 marine species of the southeastern Brazilian coast and 22 freshwater species from the Central Amazon basin. More active marine species displayed in general higher concentrations of SOD and CAT in the liver and blood, compared to those of sedentary or bottom-dwelling species. AD status in marine fish m ay be related to the oxygen consumption of the tissues and of the whole organism, while in freshwater AD may be related to physical and chemical characteristics of the environment rather than to activity level. As thermoconformer organisms, most fish must routinely cope with environmental temperature changes and, consequently, with changes in their metabolic rates. The relatively high antioxidant defense levels that typify fishes, even when compared to endotherms such as birds and mammals, may protect aquatic organisms against the consequences of temperature oscillations.

Gill diffusion as a physiological mechanism for hydrogen peroxide elimination by fish.

Hydrogen peroxide is metabolized by the specific enzymatic action of catalase and glutathione peroxidase in animal tissues. The relatively low catalase and glutathione peroxidase activities found in the blood of fish may be related to the ability of gills to eliminate hydrogen peroxide into the aquatic environment. Poecilia vellifera releases hydrogen peroxide apparently by gill diffusion into the environment, resulting in a steady-state H2O2 concentration of about 0.6 microM in the surrounding water. This physiological mechanism resembles ammonia excretion by teleost fish.

Influence of season and pollution on the antioxidant defenses of the cichlid fish acará (Geophagus brasiliensis).

The livers of Geophagus brasiliensis collected from both a non-polluted site and a polluted site were analyzed for different antioxidant defenses, O2 consumption, thiobarbituric acid-reactive substance (TBARS) levels, and histological damage. Compared to controls (116.6 +/- 26.1 nmol g-1), TBARS levels were enhanced at the polluted site (284.2 +/- 25.6 nmol g-1), as also was oxygen consumption (86.6 +/- 11.3 and 128.5 +/- 9.8 micromol O2 min-1 g-1, respectively). With respect to enzymatic antioxidants, increased catalase activities (8.7 +/- 1.3 and 29.2 +/- 2.4 mmol min-1 g-1, respectively), unchanged superoxide dismutase activities (767.2 +/- 113.3 and 563.3 +/- 70.2 U g-1, respectively), and diminished glutathione S-transferase activities (29.0 +/- 3.2 and 14.9 +/- 3.2 micromol min-1 g-1, respectively) were detected. Reduced glutathione (1.91 +/- 0.17 and 1.37 +/- 0.25 mM, respectively), oxidized glutathione (1.50 +/- 0.20 and 0.73 +/- 0.17 mM, respectively), and total glutathione (3.40 +/- 0.26 and 2.07 +/- 0.27 mM, respectively) concentrations were also below control values at the polluted site. Nevertheless, the observed ethoxyresorufin-O-deethylase activities (1.34 +/- 0.11 and 16.7 +/- 0.21 pmol min-1 mg-1, respectively) showed enhanced values at the polluted site. The main histological damage observed in the hepatocytes from fish collected at the polluted site was characterized by heavy lipid infiltration. Fish collected at the end of spring showed higher O2 consumption, higher superoxide dismutase and glutathione S-transferase activities, and higher total and oxidized glutathione concentrations compared to the beginning of autumn. No seasonal changes were observed in catalase activities, glutathione or TBARS levels. Fish chronically exposed to relatively high pollution levels seem to be unable to set up adequate antioxidant defenses, probably due to severe injury to their hepatocytes. The higher antioxidant defenses found at the end of spring are probably related to the enhanced activities during high temperature periods in thermoconforming organisms.

Proinflammatory and oxidative stress markers in patients submitted to Roux-en-Y gastric bypass after 1 year of follow-up.

BACKGROUND/OBJECTIVES: This study examined the effect of weight loss after 3, 6 and 12 months of Roux-en-Y Gastric Bypass (RYGB) on energy intake and on several biomarkers of oxidative stress such as levels of vitamin C, beta-carotene, vitamin E (diet/blood), nitric oxide metabolites (NOx), myeloperoxidase (MPO), thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) and activity of catalase (CAT). SUBJECTS/METHODS: Study with a control group (CG), assessed once, and a bariatric group (BG) assessed at the basal period as well as at 3, 6 and 12 months post-surgery; both groups were composed of 5 men and 31 women (n=36). Age was 38.7 ± 9.4 and 39.6 ± 9.2 years old and body mass index (BMI) was 22.2 0 ± 2.1 and 47.6 ± 9.1 kg/m(2), respectively. The variance measure quoted was SEM. RESULTS: The body weight at 12 months was 35.8 ± 1.0% (P<0.001) lower than that of the basal period. At the basal period BG showed higher levels of NOx (P=0.007) and TBARS (P<0.001) and lower levels of vitamins C and E (P<0.001) compared with CG. After 3 months the activity of MPO was decreased (P<0.001). Six months after surgery GSH levels were decreased (P=0.037), whereas CAT activity was increased (P=0.029). After 12 months levels of NOx (P=0.004), TBARS (P<0.001), beta-carotene (P<0.001) and vitamin E (P<0.001) were decreased, whereas those of vitamin C (P<0.001) were increased compared with controls. CONCLUSION: RYGB followed by a daily vitamin supplement apparently attenuated pro-inflammatory and oxidative stress markers 1 year after surgery, but additional antioxidant supplementation appears necessary.

Time-course variations of DNA damage and biomarkers of oxidative stress in tilapia (Oreochromis niloticus) exposed to effluents from a swine industry.

DNA damage (Comet assay), lipoperoxidation levels (TBARS), and several biomarkers of oxidative stress such as catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione S-transferase (GST), and contents of reduced (GSH) and total (TG) glutathione were measured in liver and blood (Comet assay) of tilapia (Oreochromis niloticus) exposed for 7, 15, 30 (subchronic exposure), 60, and 90 days (chronic exposure) to two treatment lagoons of a swine-processing plant, the first an anaerobic lagoon and the second a final treatment lagoon. After the 15th day, TBARS increased in fish exposed to both lagoons, decreased on the 30th day, and on the 90th day remained similar to controls. Fish exposed subchronically and chronically to both effluents showed consistently greater DNA damage. The CAT and GPx activities showed similar profiles and were induced only during the first week and during the first and second months. GST activity was induced throughout the experimental period. On the other hand, GR activities showed inverted profiles, with progressively decreased activities in the liver of fish exposed to the anaerobic lagoon, and progressively increased activities in fish exposed to the final lagoon. GSH showed higher contents in liver after 60 and 90 days of exposure to the final lagoon. GSSG contents were higher in fish exposed to the final lagoon throughout the experimental period. After 15 days, tilapia exposed to both lagoons showed enhanced total glutathione contents. The hepatic antioxidant system and biomarkers of oxidative stress such as DNA fragmentation and TBARS contents of tilapia exposed to both lagoons presented biphasic profiles. These changes in the antioxidant status also indicate that the industrial treatment is not adequate to avoid damaging environmental effects.

Evaluation of remediation of coal mining wastewater by chitosan microspheres using biomarkers.

Acidic mine waters have a marked influence on the surrounding environment and pose a serious threat through long-term environmental degradation. Therefore, it is important to improve and monitor water quality with the aim of decreasing the hazard presented by this effluent emission. The aim of this work was to evaluate the remediation of mining wastewater effluents by chitosan microspheres using biomarkers of exposure and effect. DNA damage (Comet assay) and several biomarkers of oxidative stress, such as lipoperoxidation levels (TBARS), superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) activities, and contents of reduced glutathione (GSH), were measured in blood and liver of tilapia (Oreochromis niloticus) exposed for 7, 15, and 30 days to dechlorinated tap water, 10% coal mining wastewater (CMW), and coal mining wastewater treated with chitosan microspheres (RCM). The results indicate that hepatic TBARS levels were significantly higher in fish exposed to CMW after 7, 15, and 30 days (100%, 86%, and 63%, respectively), and after remediation there was no significant difference in relation to the control group. Hepatic GSH concentrations were lower than control values for CMW after 7 and 15 days of exposure (34% decrease at both times), and this concentration was normalized by treatment with chitosan. SOD showed increased activity in liver after 15 and 30 days of exposure, 30% and 36%, respectively, and in fish exposed to RCM there was no change in this activity compared with the control group. Increased CAT activity in liver was observed during all experimental periods in fish exposed to CMW (46%, 50%, and 56% at 7, 15, and 30 days, respectively) compared with the control or treated-water groups. The highest increase in hepatic GST activity (106%) was observed only in fish exposed to CMW for 30 days. There was an increase in DNA damage in liver (50% at 7 and 15 days) and blood (79%, 77%, and 48% at 7, 15, and 30 days, respectively) after exposure to CMW. In contrast, the fish exposed to wastewater treated with chitosan microspheres exhibited DNA fragmentation indexes similar to the control group. The results obtained indicate the use of oxidative stress biomarkers as useful tools for the toxicity evaluation of coal mining effluents and also suggest that chitosan microspheres may be used as an alternative approach for remediation of coal mining wastewaters.

Erythrocytic nucleoside triphosphates in marine fish.

1. Whole blood purine nucleotides were determined in 19 species of selachians and 27 species of marine teleosts. Concomitant ATP and GTP were revealed inside the erythrocytes of almost all species studied. 2. ATP seems to be the main potential modulator of oxygen affinity in rays, sharks and teleosts, and GTP was not detected in only two teleost species. 3. The mean erythrocytic NTP concentration and the ratio between NTP and intraerythrocytic Hb concentrations in rays were the lowest (2.3 mM and 0.6 respectively), increasing in sharks (3.8 mM and 1.0), and further in teleosts (5.9 mM and 2.1). 4. The intraerythrocytic phosphate contents probably reflect different adaptative strategies associated with the fish habits and habitats, and with the Root effect expression.

Comparative hematology in marine fish.

1. A comparative study involving 80 species (14 ray, 14 shark and 52 teleost species) of marine fish found at the southeastern Brazilian coast is presented. 2. Active species displayed higher values for all hematological parameters studied when compared to the less active forms. 3. Mean values of hematocrit, hemoglobin concentration and red blood cell counts increased according to the sequence: rays, sharks, teleosts. 4. As a group, cartilaginous fish blood displayed larger and fewer erythrocytes containing more hemoglobin than teleosts; mean cell hemoglobin concentration was significantly higher in rays and sharks than in teleosts. 5. For all but the hemoglobin concentration, the hematological values studied revealed a marked contrast between bony and cartilaginous fishes which suggests distinct ways to accomplish their oxygen demands.

Comparison between the antioxidant status of terrestrial and diving mammals.

Many diving mammals are known for their ability to deal with nitrogen supersaturation and to tolerate apnea for extended periods. They are all characterized by high oxygen-carrying capacity in blood together with high oxygen storage in their muscle mass due to large myoglobin concentrations. The above properties theoretically also imply a high tissue antioxidant defenses (AD) to counteract reactive oxygen species (ROS) generation associated with the rapid transition from apnea to reoxygenation. Different enzymatic (superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase, and glutathione S-transferase), and non-enzymatic (levels of glutathione) AD as well as cellular damage (thiobarbituric acid-reactive substances contents, as a measure of lipoperoxidation) were measured in blood samples obtained from anesthetized animals, and also in blood obtained from recently dead diving mammals, and compared to some terrestrial mammals (n=5 in both groups). The results confirmed that diving mammals have, in general, higher antioxidant status compared to non-diving mammals. Apparently, to avoid exposure of tissues to changing high oxygen levels, and therefore to avoid an oxidative stress condition related to antioxidant consumption and increased ROS generation, diving mammals possess constitutive high levels of antioxidants in tissues. These data are in agreement with short-term AD adaptations related to torpor and to animals that experience large daily changes in oxygen consumption. These data are similar to the long-term adaptations of animals that undergo hibernation, estivation, freezing-thawing and dehydration-rehydration processes. In summary, animals that routinely face high changes in oxygen availability and/or consumption seem to show a general strategy to prevent oxidative damage by having either appropriate high constitutive AD and/or the ability to undergo arrested states, where depressed metabolic rates minimize the oxidative challenge.

Influence of season and pollution on the antioxidant defenses of the cichlid fish acará (Geophagus brasiliensis)

The livers of Geophagus brasiliensis collected from both a non-polluted site and a polluted site were analyzed for different antioxidant defenses, O2 consumption, thiobarbituric acid-reactive substance (TBARS) levels, and histological damage. Compared to controls (116.6 + or - 26.1 nmol g-1), TBARS levels were enhanced at the polluted site (284.2 + or - 25.6 nmol g-1), as also was oxygen consumption (86.6 + or - 11.3 and 128.5 + or - 9.8 µmol O2 min-1 g-1, respectively). With respect to enzymatic antioxidants, increased catalase activities (8.7 + or - 1.3 and 29.2 + or - 2.4 mmol min-1 g-1, respectively), unchanged superoxide dismutase activities (767.2 + or - 113.3 and 563.3 + or - 70.2 U g-1, respectively), and diminished glutathione S-transferase activities (29.0 + or - 3.2 and 14.9 + or - 3.2 µmol min-1 g-1, respectively) were detected. Reduced glutathione (1.91 + or - 0.17 and 1.37 + or - 0.25 mM, respectively), oxidized glutathione (1.50 + or - 0.20 and 0.73 + or - 0.17 mM, respectively), and total glutathione (3.40 + or - 0.26 and 2.07 + or - 0.27 mM, respectively) concentrations were also below control values at the polluted site. Nevertheless, the observed ethoxyresorufin-O-deethylase activities (1.34 + or - 0.11 and 16.7 + or - 0.21 pmol min-1 mg-1, respectively) showed enhanced values at the polluted site. The main histological damage observed in the hepatocytes from fish collected at the polluted site was characterized by heavy lipid infiltration. Fish collected at the end of spring showed higher O2 consumption, higher superoxide dismutase and glutathione S-transferase activities, and higher total and oxidized glutathione concentrations compared to the beginning of autumn. No seasonal changes were observed in catalase activities, glutathione or TBARS levels. Fish chronically exposed to relatively high pollution levels seem to be unable to set up adequate antioxidant defenses, probably due to severe injury to their hepatocytes. The higher antioxidant defenses found at the end of spring are probably related to the enhanced activities during high temperature periods in thermoconforming organisms.

Fish antioxidant defenses: a comparative approach

There are few comparative studies of vertebrate antioxidant defenses (AD) in the literature. Enzymatic (superoxide dismutase, SOD, and catalase, CAT) and non-enzymatic alpha-tocopherol, beta-carotene, ubiquinol(10) and blood glutathione) antioxidant defenses were investigated in the liver and blood of 37 fish species, 15 marine species of the southeastern Brazilian coast and 22 freshwater species from the Central Amazon basin. More active marine species display in general higher concentrations of SOD and CAT in the liver and blood, compared to those of sedentary or bottom-dwelling species. AD status in marine fish may be related to the oxygen consumption of the tissues and of the whole organism, while in freshwater fish AD may be related to physical and chemical characteristics of the environment rather than to activity level. As thermoconformer organisms, most fish must routinely cope with environmental temperature changes and, consequently, with changes in their metabolic rates. The relatively high antioxidant defense levels that typify fishes, even when compared to endotherms such as birds and mammals, may protect aquatic organisms against the consequences of temperature oscillations.

Gill diffusion as a physiological mechanism for hydrogen peroxide elimination by fish

Hydrogen peroxide is metabolized by the specific enzymatic action of catalase and glutathione peroxidase in animal tissues. The relatively low catalase and glutathione peroxidase activities found in the blood of fish may be related to the ability of gills to eliminate hydrogen peroxide into the aquatic environment. Poecilia vellifera releases hydrogen peroxide apparently by gill diffusion into the environment, resulting in a steady-state H2O2 concentration of about 0.6 µM in the surrounding water. This physiological mechanism resembles ammonia excretion by teleost fish