Gülüzar Atli

Effects of heavy metals (Cd, Cu, Cr, Pb, Zn) on fish glutathione metabolism

The glutathione metabolism contains crucial antioxidant molecules to defend the organisms against oxidants. Thus, the aim of this study was to investigate the response of the glutathione metabolism in the liver of freshwater fish Oreochromis niloticus exposed to metals (Cu, Cd, Cr, Pb, Zn) in different periods. Fish were exposed to metals (as 1xa0μg/mL) individually for 1, 7, and 14xa0days and subsequently antioxidant enzymes (glutathione peroxidase, GPX; glutathione reductase, GR and glutathione S-transferase, GST) and glutathione levels (total glutathione, tGSH; reduced glutathione, rGSH; oxidized glutathione, GSSG and GSH/GSSG ratios) in the liver were measured. There was no fish mortality during the experiments, except Cu exposure. The antioxidant enzymes responded differently to metal exposures depending on metal types and exposure durations. GPX activity increased only after Cd exposure, while GST activity increased following 7xa0days of all metal exposures. However, GR activity did not alter in most cases. Total GSH and GSH/GSSG levels generally decreased, especially after 7xa0days. Data showed that metal exposures significantly altered the response of antioxidant system parameters, particularly at day 7 and some recovery occurred after 14xa0days. This study suggests that the response of antioxidant system could help to predict metal toxicity in the aquatic environments and be useful as an “early warning tool” in natural monitoring studies.

Essential metal (Cu, Zn) exposures alter the activity of ATPases in gill, kidney and muscle of tilapia Oreochromis niloticus

An acute (96xa0h—0.1, 0.5, 1.0, 1.5xa0μg/ml) and chronic (up to 30xa0days—0.05xa0μg/ml) protocols of Cu and Zn were applied to freshwater fish Oreochromis niloticus to investigate these essential metal effects on the activities of gill, kidney and muscle Na+/K+-ATPase, Mg2+-ATPase and Ca2+-ATPase. In vitro effects of both metals (20xa0min—0.1, 0.5, 1.0, 1.5xa0μg/ml) were also measured to be able to compare both exposure routes. Data showed that ATPase activities, in general, decreased following all the exposure conditions, though there were some increases especially in Mg2+-ATPase activity. Among the enzymes, Na+/K+-ATPase and Ca2+-ATPase appeared to be more sensitive than Mg2+-ATPase to the metals. The data also indicated that effects of Cu on ATPase activity in the tissues of O. niloticus were stronger than the effects of Zn, possibly due to higher toxic effects of Cu. In vivo and in vitro exposures of metals showed similar trends with a few exceptions, especially in the gill. Variability of ATPase activity is determined by tissue type, metal species, concentration and duration. This work showed that even essential metals can alter significantly activities of ATPases in fish and thus suggests using them as a sensitive biomarker in metal contaminated waters.

Investigations on the osmoregulation of freshwater fish (Oreochromis niloticus) following exposures to metals (Cd, Cu) in differing hardness.

Hardness is one of the most important factors in water chemistry as it affects fish physiology and metal toxicity. The aim of this study was to investigate osmoregulatory responses in the Nile tilapia Oreochromis niloticus exposed to copper and cadmium (1.0μg/mL) in soft water (SW) (hardness 80mg CaCO3/L and conductivity 1.77mS/cm) and hard water (HW) (hardness 320mg CaCO3/L and conductivity 5.80mS/cm) for 0, 1, 7 and 14 days. Following the exposures, Na(+)/K(+)-ATPase activity, ion and Cu levels in the gill, kidney and intestine were measured. There was no fish mortality within 14 days, except Cu exposure in SW which killed all fish between 8 and 12 days. Generally, Na(+)/K(+)-ATPase activity was altered by both metal exposures in the gill and kidney as it increased in HW condition, but decreased in SW condition. There were also alterations in Na(+)/K(+)-ATPase activity in the intestine as its activity generally decreased. Data, in general, showed that Cd was more effective on Na(+)/K(+)-ATPase activity comparing to Cu. However, ion levels altered mainly in the kidney and intestine. Tissue metal accumulation was higher in fish tissues from SW condition comparing to HW condition. Data represented here showed that the effects of metals differed in differing water hardness. This suggests that special attention should be paid to the water chemistry when natural monitoring studies are carried out. This study also suggests that the response of osmoregulation system of fish may be a sensitive indicator under stressful conditions in different natural waters.

Alterations in the serum biomarkers belonging to different metabolic systems of fish (Oreochromis niloticus) after Cd and Pb exposures.

The serum of vertebrates including human gives sufficient data about the current health status of organism in concern. Biomarkers have gained importance in evaluation of data from biological monitoring studies. In this study, freshwater fish Oreochromis niloticus were individually exposed Cd or Pb using acute (10μM, 2 d) and chronic (20μM, 20 d) exposure protocols to investigate the alterations in serum parameters (ALP, ALT, AST, LDH, lipase, glucose, protein, cholesterol and triglyceride) and response of the endocrine system functioning in different axis, namely HPI (cortisol), HPT (TSH, T3, T4), gonadal (LH, FSH) and prolactin. Data showed that except LH, the levels of all hormones decreased significantly following exposure to Cd and Pb both in acute and chronic exposures. In acute exposures, the activity of ALP and levels of cholesterol and triglyceride decreased significantly following both Cd and Pb exposures, while glucose levels increased only after Cd exposure. In chronic exposures, both metal exposures caused significant decreases in ALP activity and levels of cholesterol and triglyceride, though there were increases in glucose level after Cd exposure and AST, ALT and LDH levels after Pb exposure. Data emphasized the importance of biomarker selection and multiparameter usage of relevant systems in ecotoxicological research to achieve proper evaluation of environmental data.

Effects of fish size on the response of antioxidant systems of Oreochromis niloticus following metal exposures

The size of a fish is an important factor in its physiology, and metal uptake is affected by animal physiology. In this study, small and large tilapias (Oreochromis niloticus) differing approximately twofold in length and fivefold in weight were compared for their antioxidant response. Both groups were exposed to Cu or Cr (1.0xa0μg/mL) in a freshwater (−80xa0mg CaCO3/L, conductivity 1.77xa0mS/cm) using 2 exposure protocols (20xa0μM for 48xa0h and 10xa0μM for 6xa0days). Following the exposures, the antioxidant enzyme activities (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX; glutathione reductase, GR and glutathione S-transferase, GST) and glutathione (GSH) levels were measured in the liver of fish. Results showed that small fish was affected from exposure conditions much more than large ones as their antioxidant parameters significantly decreased even in controls. Metal exposures of small fish caused significant increases in SOD and CAT activity in acute Cu or Cr exposures. Subchronic Cr exposure of small fish also caused significant increases in CAT, GPx and GST activities, while there was no significant change in Cu-exposed ones. Large fish, however, showed different antioxidant responses as their levels mostly decreased. This study demonstrated that the response of antioxidant system in the liver of tilapia varied in relation to fish sizes and emphasized using different size groups in environmental monitoring and also in evaluation of fish biomarkers.

The effects of increased freshwater salinity in the biodisponibility of metals (Cr, Pb) and effects on antioxidant systems of Oreochromis niloticus

Anthropogenic activities can increase the salinity of freshwaters and this may cause stress for fish and affect metal bioavailability. Oxidative stress biomarkers are of great interest due to their responses to environmental stressors which provide valuable data for biological monitoring of aquatic pollution. Thus, the individual and combined effects of salinity and metals (Cr, Pb) were investigated in the liver of freshwater fish Oreochromis niloticus in the present study. Fish were exposed to salinity (2 and 8 ppt) alone and salinity+metal (1 μg/mL Pb and Cr) combination exposures for 0, 1, 7 and 14 days and subsequently antioxidant enzymes (superoxide dismutase, SOD; glutathione peroxidase, GPX; glutathione reductase, GR and glutathione S-transferase, GST) activities and glutathione (GSH) levels in the liver were measured. Data showed that all the parameters varied in relation to metal species, exposure durations and salinity levels. Profound alterations on the measured parameters were detected at the lower salinity compared to the higher one. Salinity increase effectively stimulated the antioxidant parameters. The effects of salinity and metals on the measured parameters increased as the exposure duration prolonged. SOD was the most affected antioxidant parameter from both salinity and metals. Because metal and salinity stresses affect fish antioxidant system, this work suggests that the chemistry of freshwaters should be taken into account in natural monitoring for metal contamination in the field.

The Effects of Salinity and Salinity+Metal (Chromium and Lead) Exposure on ATPase Activity in the Gill and Intestine of Tilapia Oreochromis niloticus

Freshwater organisms are highly sensitive to increases in salinity because they causes serious osmoregulation problems. Salinity of inland waters can be increased as a result of anthropogenic activities. In this study, freshwater fish Oreochromis niloticus were exposed individually to increased salinities (0, 2, and 8xa0ppt) alone and salinity+metal [1xa0μg/mL chromium (Cr) or lead (Pb) exposure at 2 and 8xa0ppt] exposures for different time periods (1, 7, and 14xa0days) to investigate the response of Na+/K+-ATPase and Mg2+-ATPase in the osmoregulatory tissues (gill and intestine). Results showed that enzyme activity varied depending on salinity, tissue, metal, and exposure duration. Metal levels in controls and salinity-exposed groups were lower than the detection limit, although significant Cr and Pb accumulation occurred in the salinity+metal combination groups. In salinity-exposed groups, there were increasing trends in the enzyme activity, whereas there were decreasing trends in the metal+salinity groups. Gill ATPases were more affected by the exposure conditions compared with intestine ATPases. Results showed that salinity+metal exposure both played significant roles on ATPase activities in the osmoregulatory tissues, although the alterations in the activity were mostly insignificant supporting compensation mechanisms. Results also suggest that the osmoregulation of freshwater fish should be investigated in toxicity- monitoring programs in inland waters.

Metals (Ag+, Cd2+, Cr6+) affect ATPase activity in the gill, kidney, and muscle of freshwater fish Oreochromis niloticus following acute and chronic exposures

Freshwater fish Oreochromis niloticus were individually acutely exposed to different concentrations (0, 0.1, 0.5, 1.0, and 1.5 μg/mL) of Cd2+, Cr6+, and Ag+ for 96 h and 0.05 μg/mL concentration of the same metals for different periods (0, 5, 10, 20, and 30 days) chronically. Following each experimental protocol, Na+/K+‐ATPase, Mg2+‐ATPase, and Ca2+‐ATPase activities were measured in the gill, kidney, and muscle of O. niloticus. In vitro experiments were also performed to determine the direct effects of metal ions (0, 0.1, 0.5, 1.0, and 1.5 μg/mL) on ATPases. Except Ag+, none of the metals caused fish mortality within 30 days. Silver killed all the fishes within 16 days. Metal exposures generally decreased Na+/K+‐ATPase and Ca2+‐ATPase activities in the tissues of O. niloticus, although there were some fluctuations in Mg2+‐ATPase activity. Ag+ and Cd2+ were found to be more toxic to ATPase activities than Cr6+. It was also observed that metal efficiency was higher in the gill than in the other tissues. Results indicated that the response of ATPases varied depending on metals, exposure types, and tissues. Because ATPases are sensitive to metal toxicity, their activity can give valuable data about fish physiology. Therefore, they may be used as a sensitive biomarker in environmental monitoring in contaminated waters.

Response of ATPases in the osmoregulatory tissues of freshwater fish Oreochromis niloticus exposed to copper in increased salinity

An increase in salinity of freshwater can affect the physiology and metal uptake in fish. In the present study, Nile tilapia Oreochromis niloticus were exposed to copper (1.0xa0mg/l) in increased salinities (2, 4, and 8 ppt) for 0, 1, 3, 7, and 14xa0days. Following the exposures, the activities of Na+/K+-ATPase, Mg2+-ATPase, and Ca2+-ATPase were measured in the gill, kidney, and intestine to evaluate the changes in osmoregulation of fish. Results showed that increases in salinity and Cu exposure of fish significantly altered the ATPase activities depending on the tissue type, salinity increase, and exposure durations. Salinity-alone exposures increased Na+/K+-ATPase activity and decreased Ca2+-ATPase activity. Na+/K+-ATPase activity decreased following Cu exposure in 2 and 4 ppt salinities, though the activity increased in 8 ppt salinity. Ca2+-ATPase activity decreased in the gill and intestine in all salinities, while the activity mostly increased in the kidney. However, there were great variations in Mg2+-ATPase activity following exposure to salinity alone and salinity+Cu combination. Cu accumulated in the gill and intestine following 14xa0days exposure and accumulation was negatively correlated with salinity increase. Data indicated that ATPases were highly sensitive to increases in salinity and Cu and might be a useful biomarker in ecotoxicological studies. However, data from salinity increased freshwaters should carefully be handled to see a clear picture on the effects of metals, as salinity affects both metal speciation and fish osmoregulation.

Characterization and response of antioxidant systems in the tissues of the freshwater pond snail (Lymnaea stagnalis) during acute copper exposure.

The response of enzymatic (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX and glutathione reductase, GR) and non-enzymatic responses (glutathione, GSH, oxidized glutathione, GSSG and GSH/GSSG) against acute Cu toxicity (2-90μg/mL for 48h) in different tissues of Lymnaea stagnalis were measured. Incubation conditions for enzymatic activity measurements were optimized for L. stagnalis tissues. Three examined tissues, the hepatopancreas, the foot muscle and the mantle, exhibited variable responses in antioxidant parameters as a function of Cu concentrations. The most responsive antioxidant enzymes were GPX and CAT while GR appeared less sensitive. In general antioxidant enzymes at higher Cu concentrations though GSH levels at lower Cu concentrations exhibited the greatest changes in hepatopancreas and foot muscle, respectively. All antioxidant enzymes except GR increased after exposure to the highest Cu concentration in mantle. Total and reduced GSH increased in hepatopancreas but decreased with GSH/GSSG ratios at all Cu concentrations in foot muscle. The present results show that antioxidants respond to acute Cu exposure at concentrations as low as 2μg Cu/L in adult L. stagnalis with variable responses in different tissues. Antioxidants both including enzymatic and non-enzymatic parameters may account, in part, for the high tolerance to acute metal exposure observed in adult L. stagnalis and could form suited biomarkers to evaluate the metal exposure and toxicity in aquatic environment even at relatively low level short term exposure.