Changes in energy reserves and responses of some biomarkers in freshwater mussels exposed to metal-oxide nanoparticles.

In this study, responses of various biomarkers in the digestive gland and foot muscle of freshwater mussels (Unio tigridis) were investigated following exposure to Al2O3, CuO and TiO2 nanoparticles (NPs) for 14 days at different concentrations (0, 1, 3 and 9 mg NP/L). Mussels were fed on unicellular algae (Chlorella vulgaris) cultured in the laboratory. NP exposures caused significant increases (p < 0.05) in the levels of total glutathione (GSH), reduced-glutathione (rGSH), oxidized-glutathione (GSSG) and malondialdehyde (MDA) in the digestive gland. Oppositely, there were significant (p < 0.05) decreases in acetylcholinesterase activity in the foot muscles. Total energy reserves of the digestive gland and foot muscle significantly (p < 0.05) decreased, but only at the highest NP exposures. Nevertheless, NP exposures did not alter (p > 0.05) the algae filtering capacity of mussels. This study demonstrated that the biomarkers belonging to different metabolic systems responded to NP exposures, suggesting their usage in the monitoring studies for freshwater systems.

Responses of oxidative stress biomarkers of freshwater fish (Oreochromis niloticus) exposed to Cr6+, Hg2+, Ni2+ and Zn2+ in differing calcium levels.

Freshwaters from different geographical locations show different hardness, affecting metal uptake and toxicity in fish. The most important ion that determines water hardness is calcium. In this study, acute and chronic effects of metals on the oxidative stress biomarkers in the liver of freshwater fish (Oreochromis niloticus) were investigated in differing Ca2+ (30, 60 and 120 mg Ca2+/L) levels. Fish were exposed to Cr6+, Ni2+ and Zn2+ (30 µM) and Hg2+ (0.3 µM) for 3 days in acute experiments, while they were exposed to Cr6+, Ni2+ and Zn+2 (10 µM) and Hg2+ 0.03 µM) for 30 days in chronic experiments. Data showed that the oxidative stress biomarkers significantly (p < 0.05) altered after metal exposures at all calcium levels, though there was no significant change (p > 0.05) among calcium controls. In both acute and chronic exposures, catalase CAT) and superoxide dismutase (SOD) activities increased significantly, while glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione S-transferase (GST) activities decreased. There were significant decreases in total glutathione (GSH) levels in acute exposures, though GSH levels increased in chronic exposures. Malondialdehyde (MDA) levels significantly increased in both durations. The highest significant alterations in the biomarkers occurred at the lowest Ca2+ levels. GPX and GST were found to be the most sensitive enzymes in all exposures and the least alterations in biomarker response occurred in fish exposed to Ni2+. This study demonstrated that calcium levels (hardness) were important factors in the evaluation of metal toxicity for freshwater fish.

Accumulation and Elimination of Cadmium by the Nile Tilapia (Oreochromis niloticus) in differing Temperatures and Responses of Oxidative Stress Biomarkers.

The aims of this study were to investigate the accumulation (15 days) and elimination (15 and 30 days) of cadmium (Cd) in the liver, gill, kidney and muscle of Oreochromis niloticus following exposures to different concentrations (1 and 2 mg/L) of Cd at different water temperatures (20, 25, 30 oC). Additionally, responses of oxidative stress biomarkers (superoxide dismutase, SOD; catalase CAT; glutathione peroxidase, GPx and malondialdehyde, MDA) of the liver were determined following Cd exposures. In accumulation period, Cd levels increased significantly in all the tissues at all temperatures and tissue accumulation order was kidney > liver > gill. All tissues, except the muscle, accumulated Cd in relation to exposure concentrations and water temperatures. There was no measurable level of Cd accumulation in the muscle, except in fish exposed to 2 mg Cd/L at 30 oC. Likewise, elimination of Cd from the tissues also increased in depends on periods and water temperatures. The order of Cd elimination from the tissues was gill > liver > kidney. The oxidative stress biomarkers also responded to both Cd exposure and temperature increases. The activities of antioxidant enzymes such as SOD, CAT, GPx and MDA levels in the liver increased in relation to increases in Cd concentrations and water temperatures.

Salinity and/or nanoparticles (Al2O3, TiO2) affect metal accumulation and ATPase activity in freshwater fish (Oreochromis niloticus).

The osmoregulation system of freshwater fish is sensitive to salinity increase in water. There is no satisfactory data to our knowledge on the accumulation of metal-oxide nanoparticles (NPs) in tissues of O. niloticus and their effects on ATPases (Na,K-ATPase, Mg-ATPase, Ca-ATPase) in differing salinities. Thus, this study investigated the effects of salinity (0 and 10 ppt) and Al2O3 and TiO2 NPs (1 and 10 mg NPs/L) on the response of ATPases in acute (2 days) and chronic (20 days) durations. Data showed that nanoparticles accumulated in the tissues of fish, gill tissues having the highest levels of Al and Ti in both acute and chronic durations. Interestingly, the higher salinity significantly increased (P < 0.05) NP accumulations in the tissues in acute exposures, whereas it significantly decreased (P < 0.05) in chronic exposures. Salinity increase caused significant decreases (P < 0.05) in ATPase activities (up to 54 %) in control fish from both exposure protocols. Likewise, NP alone exposures (up to 80 %) and salt+NP (up to 83 %) exposures generally caused significant (P < 0.05) decreases in ATPase activities compared to their controls. Similarly, salt+NP exposures also decreased ATPase activities compared to NP exposures alone. The present data demonstrated that salinity and/or NP exposures decreased ATPase activities in the gill of freshwater fish, emphasizing the possible hazardous consequences of salt inputs and NP discharges into freshwater systems.

The effects of titanium nanoparticles on enzymatic and non-enzymatic biomarkers in female Wistar rats.

Titanium dioxide (TiO2) nanoparticles (NPs) are widely used in industry, pharmacy, medicine, and food sectors. Therefore, this study deals with the effects of TiO2 NPs in female rats following oral administration in differing doses for 14 days (0, 0.5, 5, and 50 mg/kg b.w./d). The response of enzymatic biomarkers (Na,K-ATPase, Mg-ATPase, and AChE) was measured in the brain, kidney, and small intestine, while non-enzymatic biomarker levels, such as different forms of glutathione (GSH) and thiobarbituric acid reactive substances (TBARSs) were measured in the liver. The images of the tissues were obtained using a transmission electron microscope (TEM) to demonstrate TiO2 NP accumulation. Data showed that brain AChE activity decreased at all TiO2 NP doses, though brain ATPase activities increased. However, ATPase activities in the intestine and kidney did not change significantly. Levels of GSH forms did not change significantly, though there was a significant decrease in TBARS level at the highest NP dose. TEM images demonstrated that TiO2 NPs accumulated in a dose-dependent manner in the tissues. Data emphasized that the brain was the most sensitive organ against the effects of TiO2 NPs. This study suggests the need for further studies to evaluate better the toxic effects of TiO2 NPs.

Accumulation and Distribution of Nanoparticles (Al2O3, CuO, and TiO2) in Tissues of Freshwater Mussel (Unio tigridis).

Freshwater mussels are used as an effective bioindicator of metal pollution. There is no data on the accumulation of any metal-oxide nanoparticles (NPs) in tissues of Unio tigridis. Thus, this study was undertaken to investigate accumulation of Al2O3, CuO, and TiO2 NPs following exposure to different concentrations (0, 1, 3, and 9 mg/L) of NPs for 14 days. Metal concentrations in tissues were determined by ICP-MS, while NP presence was demonstrated by transmission electron microscope (TEM) images. During the experiments, mussels were fed with cultured algae (Chlorella vulgaris). TEM images demonstrated the presence of NPs in digestive gland and muscle. TEM images also suggested that NPs were taken via the lysosomes or endosomes. Highest mean concentrations (µg/g d.w.) of aluminium (76.51), copper (111.63) and titanium (113.83) occurred in the gills and followed by the digestive glands and muscles. Algae consumption of mussels did not significantly differ among controls and NP-exposed groups.

Characterization of ATPases in the gill of freshwater mussel (Unio tigridis) and effects of ionic and nanoparticle forms of aluminium and copper.

Mussels are often used to monitor the aquatic systems for different ecological aims, as they are one of the best bioindicator animals. However, the characterization of biomarkers should be known before using them in environmental monitoring and toxicology studies. There is no study to our knowledge on the characterization of Na-ATPase and Ca-ATPase in tissues of freshwater mussel (Unio tigridis). Thus, this study was undertaken to characterize the optimum working conditions of Na-ATPase and Ca-ATPase in the gill of mussels, determining the highest levels of parameters (Na+, Mg2+, Ca2+, ATP, pH, temperature, enzyme amount, incubation time) to obtain maximum activity. The present study also aimed to investigate in vitro effects of ionic and nanoparticle (Al2O3, CuO) forms of aluminium and copper (0, 30, 90, 270 µg/L) on the activities of Na-ATPase and Ca-ATPase. Data showed that there was no ouabain-sensitive ATPase activity in the gill up to 10 mM ouabain concentrations. Na-ATPase and Ca-ATPase activities in the gill of control mussels were 5.124 ± 0.373 and 3.750 ± 0.211 µmol Pi/mg pro./h, respectively. Exposure to different concentrations of nanoparticles did not alter significantly (P > 0.05) the activities of Na-ATPase and Ca-ATPase in vitro, whereas the same concentrations of ionic aluminium and copper significantly decreased (P < 0.05) the enzyme activities. Data emphasized that there were different modes of action between ionic and nanoparticle forms of aluminium and copper. Data also suggested that in vivo studies should also be carried out to estimate better the effects of nanoparticle and ionic forms of metals on ATPases of U. tigridis.

Antioxidant system biomarkers of freshwater mussel (Unio tigridis) respond to nanoparticle (Al2O3, CuO, TiO2) exposures.

PURPOSE: Mussels are known as the natural filters of the aquatic systems and are accepted as one of the best bioindicator organism. There is no data on the response of Unio tigridis to metal-oxide nanoparticle (NP) exposures. This study aimed to investigate the response of the antioxidant enzymes of U. tigridis following exposure to NPs. MATERIALS AND METHODS: The mussels were exposed to different concentrations (0, 1, 3, 9 mg NP/L) of Al2O3, CuO and TiO2 NPs for 14 days and subsequently, the activities of CAT (catalase), SOD (superoxide dismutase), GPx (glutathione peroxidase), GST (glutathione S-transferase) and GR (glutathione reductase) were measured in the gill and digestive gland. Mussels were fed with cultured unicellular algae (Chlorella vulgaris) during experiments. RESULTS: Data showed that algae consumptions of mussels were not significantly (p>0.05) altered by NPs. However, all enzyme activities in the digestive gland and gill altered significantly (p<0.05) after NP exposures. The activities of CAT and SOD decreased, while the activities of enzymes belonging to glutathione metabolism (GPx and GST) increased in both tissues. CONCLUSION: This study representing the first record on the antioxidant system response of U. tigridis toward NP exposures suggests that NP toxicity should be investigated thoroughly in organisms and some regulations must be done on their usages.

Responses of biomarkers belonging to different metabolic systems of rats following oral administration of aluminium nanoparticle.

Nanoparticle (NP) forms of aluminium oxide (Al2O3) are used in various fields such as engineering, pharmacy, medicine etc. Compounds containing aluminium oxide NPs may present toxic effects after certain thresholds. Thus, the present study was carried out to determine the effects of Al2O3 nanoparticles (Al-NPs) in rats. For this aim, different doses (0, 0.5, 5, 50 mg/kg b.w./day) of Al NP (˜40 nm) were orally administered to female rats (Rattus norvegicus var. albinus) for 14 days and the response of several biomarkers such as activities of ATPases (total ATPase, Na,K-ATPase, Mg-ATPase) and acetylcholinesterase (AChE), levels of different glutathione forms and thiobarbituric acid reactive substances (TBARS) were measured in different tissues. Additionally, tissue accumulation of Al-NPs was demonstrated by a transmission electron microscope (TEM). The images showed the presence of Al-NP aggregates in all the tissues at all doses. The sizes of NP aggregates were dependent on NP doses and it was a bit more loose in the brain than in the liver and kidney. AChE activity in the brain decreased significantly at all NP doses, whereas TBARS levels in the liver did not alter significantly at any NP dose. Although there was no significant change in ATPase activities in the intestine at any NP dose, there were significant decreases in the kidney and brain. There were some variations in the levels of total glutathione (tGSH), oxidized glutathione (GSSG) and reduced glutathione (rGSH), though these variations were not significant (P > 0.05). Likewise, the ratio of rGSH/GSSG also did not differ significantly among NP doses and control. The brain seems most affected organ following Al-NP administration. This study demonstrated that most biomarkers in the tissues of rats were affected by Al-NP, showing the signal of toxic effects and suggests further studies to understand better the effects of Al NPs, especially in their use for pharmacology.

Response of the antioxidant enzymes of rats following oral administration of metal-oxide nanoparticles (Al2O3, CuO, TiO2).

Metal-oxide nanoparticles (NPs), as a new emerging technological compound, promise a wide range of usage areas and consequently have the potential to cause environmental toxicology. In the present work, aluminum (Al2O3), copper (CuO), and titanium (TiO2) nanoparticles (NPs) were administered via oral gavage to mature female rats (Rattus norvegicus var. albinos) for 14 days with a dose series of 0 (control), 0.5, 5, and 50 (mg/kg b.w./day). Enzyme activities of the antioxidant system such as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST), and glutathione reductase (GR) in the liver were measured. Transmission electron microscope (TEM) images of the liver were taken to demonstrate NP accumulation and distribution in liver tissue. Data showed that all NPs caused some significant (P > 0.05) alterations in the activities of antioxidant enzymes. CAT activity increased after CuO and TiO2 administrations, while SOD activity decreased after Al2O3 administration. The activities of enzymes associated with glutathione (GR, GPx, GST) metabolisms were also significantly altered by NPs. GPx activity increased in rats received Al2O3, CuO NPs, while GR activity increased only by Al2O3. However, there were increases (TiO2) and decreases (CuO) in GST activity in the liver of rats. TEM images of the liver demonstrated that all NPs accumulated in the liver, even at the lowest dose. This study indicated that the antioxidant enzymes in the liver of rats were affected by all NPs, suggesting the antioxidant system of rats suffered after NP administration.

Serum biomarker levels alter following nanoparticle (Al2O3, CuO, TiO2) exposures in freshwater fish (Oreochromis niloticus).

Nanoparticles (NPs) are used in diverse field of technology and consequently are released to the environment, most ending up in water bodies. Because NPs have potential to cause adverse effects on the metabolisms of animals, the present study was carried out to help understanding their effects on fish metabolism. In this study, freshwater fish (Oreochromis niloticus) were exposed to aluminum oxide (Al2O3) (40 nm), copper oxide (CuO) (40 nm) and titanium dioxide (TiO2) (21 nm) NPs in differing concentrations (0, 1, 5, 25 mg/L) for 14 d. Following the exposures, the levels of glucose, cholesterol, triglyceride, blood urea nitrogen (BUN), creatinine, alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), Na+, K+, triiodothyronine (T3), thyroxine (T4) and cortisol in the serum of fish were measured. The results of this study showed that there were significant alterations in the levels of some serum parameters. The levels of glucose, triglyceride, BUN, creatinine, K+, cortisol and T4 in the serum were changed following NP exposures, though there was no statistical difference in the levels of cholesterol, ALT, AST, ALP, Na+ and T3. Most striking data were obtained in the levels of BUN and creatinine, as their levels increased nearly 10 folds. Transmission electron microscope (TEM) images showed NP accumulation in tissues of fish even at the lowest exposure concentration. This study emphasized that NPs are not innocent compounds and can have hazardous effects when taken in substantial levels, suggesting there must be some criteria and limits in their usage and discharge to the environment. Nevertheless, more studies are needed to understand better their toxicities in different classes of animals.

Investigations on the effects of etoxazole in the liver and kidney of Wistar rats.

Pesticides are used to protect crops and to eliminate pests, though non-target organisms such as mammals are also affected from their usage. Etoxazole (organoflourine pesticide) is an acaricide used to combat spider mites which are the parasites of various crops. The present study aims to investigate the effects of etoxazole on the level of MDA (malondialdehyde) and activities of CAT (catalase), GPx (glutathione peroxidase), and AChE (acetylcholinesterase) in liver and kidney tissues of Wistar rats (Rattus norvegicus var. albinos). Rats received etoxazole intraperitoneally with doses of 2.2, 11, and 22 mg/kg b.w./day for 21 days. Control rats received the same volume of the serum physiologic. Following etoxazole exposures, activities of CAT, GPx, and AChE in the liver and kidney of rats significantly decreased at all doses compared to control group. Oppositely, MDA levels in these tissues increased significantly at all doses following etoxazole exposures. The present study demonstrated that etoxazole, at all doses, had toxic effects in the liver and kidney parameters, suggesting their possible use as effective biomarkers in determining the toxic effects of etoxazole. This may suggest that these biomarkers could also be used as a tool to monitor pesticide-affected areas before severe toxic effects begin in non-target animals and humans.

Response of the antioxidant enzymes of the erythrocyte and alterations in the serum biomarkers in rats following oral administration of nanoparticles.

In this study, Al2O3, CuO and TiO2 nanoparticles (NPs) were administered to mature female rats (Rattus norvegicus var. albinos) via oral gavage (0, 0.5, 5, 50mg/kg b.w./day) for 14days to investigate their effects on 14 serum biomarkers and 4 antioxidant enzyme (catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase) activities in the erythrocyte. Data showed that Al2O3 did not cause any significant (P>0.05) change in the parameters, except few cases, while CuO and TiO2 caused significant alterations in antioxidant system parameters of the erythrocytes. Activities of catalase and superoxide dismutase significantly decreased in CuO and TiO2 administered rats. Oppositely, glutathione peroxidase activity increased in CuO and TiO2 administered rats. There were no significant alterations in the activity of glutathione S-transferase in the erythrocytes. Levels of glucose, cholesterol, bilirubin, triglyceride, triiodothyronine (T3), estradiol, prolactin and immunoglobulin M (IgM) in the serum altered after some of NP administrations, whereas cortisol, protein, creatinine, blood urea nitrogen (BUN), thyroxine (T4) and immunoglobulin G (IgG) levels in the serum did not change significantly after any of NP administration. There were outstanding increases in the levels of bilirubin and prolactin and decreases in the levels of triglyceride and estradiol. The present study demonstrated that the antioxidant enzymes in the erythrocyte were generally affected from copper and titanium NPs, while aluminium and copper NPs caused more significant alterations in serum biomarkers.

Characterization of antioxidant system parameters in four freshwater fish species.

The potential use of antioxidant system parameters has gained considerable interest due to their pivotal role of detoxification mechanisms in environmental studies and culture fish point of view. Fish with different ecological needs may have different antioxidant capacity and response to environmental contaminants. Thus, the optimal working conditions and specific enzyme activities (Vmax and Km) of antioxidant system parameters (Superoxide dismutase, SOD; Catalase, CAT; Glutathione peroxidase, GPX; Glutathione reductase, GR and Glutathione S-transferase, GST) and glutathione (GSH) were determined in four commonly cultured freshwater fish species (tilapia; Oreochromis niloticus, carp; Cyprinus carpio, trout; Onchorhynchus mykiss and catfish; Clarias garipienus). Data showed that optimal concentrations of different buffers, pH and specific chemicals for each enzyme and GSH were similar in most cases for all fish species, except a few differences. The highest Vmax and Km values were found in carp for GPX and GST, though these values were the highest in tilapia, catfish and trout for CAT, SOD and GR, respectively. As a conclusion, optimization assays of these parameters in different bioindicator organisms based on their physiological and ecological differences may be useful for the aquatic ecosystem biomonitoring studies and also present fundamental data for utilization in aquaculture.

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.

Low water conductivity increases the effects of copper on the serum parameters in fish (Oreochromis niloticus).

The conductivity is largely determined by ion levels in water, predominant ion being Ca(2+) in the freshwaters. For this reason, the effects of copper were evaluated as a matter of conductivity of exposure media in the present study. Thus, freshwater fish Oreochromis niloticus were exposed to copper in differing conductivities (77, 163 and 330 µS/cm), using acute (0.3 µM, 3 d) and chronic (0.03 µM, 30 d) exposure protocols. Following the exposure serum parameters of fish were measured. Data showed that there was no significant alteration (P>0.05) in serum parameters of control fish. However, activities of ALP, ALT and AST decreased significantly at the lower conductivities in chronic copper exposure, but not in acute ones. Protein levels did not differ significantly in any of the exposure conditions. However, Cu exposure at the lowest conductivity sharply increased the levels of glucose in the acute exposure, while there was no significant difference in the chronic exposure. Cholesterol levels decreased only at the lower conductivities in chronic exposure, but increased in acute exposure. Similarly, triglyceride levels increased in acute exposures and decreased in chronic exposures at the lowest conductivity. There was no change in Na(+) levels, while there was an increase in K(+) levels and a decrease in Ca(2+) level at the lowest conductivity of acute exposures. However, Cl(-) levels generally decreased at the higher conductivities of chronic exposures. There was a strong negative relationship between significant altered serum parameters and water conductivity. In conclusion, this study showed that copper exposure of fish at lower conductivities caused more toxicities, indicating the protective effect of calcium ions against copper toxicity. Data suggest that conductivity of water may be used in the evaluation of metal data from different waters with different chemical characteristics.

The effects of cyfluthrin on some biomarkers in the liver and kidney of Wistar rats.

Pesticides can cause oxidative stress resulting to deleterious effects in animal metabolisms. Cyfluthrin is a synthetic pyrethroid used worldwide to protect crops and to eliminate pests. Thus, the aim of this study was to investigate the effects of the cyfluthrin on the level of malondialdehyde (MDA) and activities of catalase (CAT), glutathione peroxidase (GPx), and acetylcholinesterase (AChE) in the liver and kidney of Wistar Albino Sprague Dawley rats (Rattus norvegicus var. albinos) following intraperitoneal treatment of cyfluthrin (1.2, 12, and 120 mg/kg b.w./day) for 21 days. Comparisons were made with two control solutions named as serum physiologic and solvent in which cyfluthrin was dissolved. CAT activity in the liver and kidney of rats did not change after the lowest cyfluthrin treatment, while its activity significantly decreased at the higher doses. In general, cyfluthrin significantly decreased the activity of GPx in the liver and kidney at all doses, while MDA levels in the liver increased at all doses. Cyfluthrin significantly decreased AChE activity in the liver of rats at all doses, while this was true at the highest dose for the kidney. This study showed that the studied biomarkers were effective in determining the toxic effects of cyfluthrin. Thus, they should be used to monitor pesticide-affected areas before untargeted animals, including humans who suffer from the use of pesticides.

Response of antioxidant system of tilapia (Oreochromis niloticus) following exposure to chromium and copper in differing hardness.

Tilapias (Oreochromis niloticus) were exposed to copper or chromium in soft water (SW) (~80 mg CaCO3/L, conductivity 1.77 mS/cm) or hard water (HW) (~320 mg CaCO3/L, conductivity 5.80 mS/cm) using 2 exposure protocols (20 µM for 48 h and 10 µM for 144 h). Following the exposures, antioxidant enzyme activities [superoxide dismutase (SOD); catalase (CAT); glutathione peroxidase; glutathione reductase; and glutathione S-transferase (GST)] and glutathione (GSH) levels were measured in the liver of fish. SOD and CAT activities of control fish kept in SW were significantly lower than control fish kept in HW. However, the other antioxidant indices (glutathione metabolism) of both control fish were unaffected from water hardness. Acute metal exposures did not alter the glutathione metabolism, whereas SOD activity in SW and CAT activity in both waters changed significantly. In subchronic duration, Cu exposure caused significant decreases in measured parameters, except for GST activity and GSH level. Similarly, GST activity and GSH level were unaffected from Cr exposure. This study showed that SOD and CAT were the most sensitive antioxidant indices, and that glutathione metabolism, in general, was not altered following metal exposures in different waters.

Effects of Cd, Zn and Cd + Zn combination on ATPase activitiy in the gill and muscle of tilapia (Oreochromis niloticus).

This study investigated the responses of Na(+)/K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase in the gill and muscle of a freshwater fish Oreochromis niloticus exposed to 1 µg/mL of Cd and Zn and their mixture for different periods (0, 7, 14, 21 and 28 days). At the end of experimental periods, the activities Na(+)/K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase in gill tissues and only Ca(2+)-ATPase activity in muscle tissues were measured. Gill Na(+)/K(+)-ATPase activity generally decreased following single metal exposures, whereas their combinations increased its activity. Gill Ca(2+)-ATPase activity decreased relative to the control at most exposure times for single exposures of Zn and Cd, as well as for the combined exposure. There was no gill Ca(2+)-ATPase activity after 28 days of exposure to Zn and Cd combined. Mg(2+)-ATPase activity was not affected significantly in gill tissue by exposure to Zn and Cd individually or in combination. Muscle Ca(2+)-ATPase activity also decreased significantly following metal exposure, but not as greatly as in the gill tissue. Tissue protein levels were mostly unaffected by metal exposures. This study showed that certain ATPases are highly sensitive to metal exposure whether the metals are essential or non essential, and suggests using gill tissue Na(+)/K(+)-ATPase and Ca(2+)-ATPase as sensitive biomarkers 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.