The Effects of Salicylic Acid on Juvenile Zebrafish Danio rerio Under Flow-Through Conditions.

The aquatic environment is becoming increasingly contaminated with pharmaceuticals. Salicylic acid (SA), which can be used individually or appear as a degradation product of the widely used acetylsalicylic acid was chosen for testing. Juvenile zebrafish Danio rerio were subjected to OECD test No. 215 (fish, juvenile growth test) with salicylic acid concentrations of 0.004; 0.04; 0.4; 4 and 40 mg/L. Specific growth rate (SGR), histological changes, and parameters of oxidative stress were evaluated. SA had no effects on histological changes, SGR, glutathione reductase, and lipid peroxidation. Increased catalytic activity of GPx was found at 0.04 mg/L compared to control, increased catalytic activity of catalase was found at 0.04 and 4 mg/L compared to control, and increased catalytic activity of glutathione-S-transferase was found at 0.004 and 0.04 mg/L compared to control (P < 0.05). Juvenile zebrafish turned out to be relatively insensitive to both environmentally relevant (0.004 mg/L) and higher concentrations of salicylic acid.

The effects of ciprofloxacin on early life stages of common carp (Cyprinus carpio).

The authors performed a toxicity test with ciprofloxacin in fertilized eggs of common carp according to guideline 210 of the Organisation for Economic Co-operation and Development. The tested concentrations were 1 µg L(-1) , 100 µg L(-1) , 500 µg L(-1) , 1000 µg L(-1) , and 3000 µg L(-1) . Accelerated hatching was found in all groups exposed to ciprofloxacin, but significant growth reduction was found only in the group exposed to the highest concentration (3000 µg L(-1) ). Increased numbers of macroscopic morphological anomalies were observed on day 6 of the test (after hatching). The highest numbers of macroscopic morphological anomalies were observed in the groups of free embryos and larvae exposed to ciprofloxacin concentrations of 100 µg L(-1) , 500 µg L(-1) , 1000 µg L(-1) , and 3000 µg L(-1) (20-23% of tested samples). A gradual decrease in glutathione S-transferase activity was detected in all experimental groups exposed to ciprofloxacin, but significant differences (p < 0.01) were found only in groups treated with 500 µg L(-1) and 3000 µg L(-1) . Glutathione peroxidase and catalase exhibited increased activity in most of the tested concentrations (p < 0.01 and <0.05, respectively), whereas decreased glutathione reductase activity was found in the groups exposed to ciprofloxacin concentrations of 500 µg L(-1) and 3000 µg L(-1) (p < 0.05). The concentration of thiobarbituric acid-reactive substances was significantly lower (p < 0.01) in all experimental groups exposed to ciprofloxacin. The lowest-observed-effect concentration of ciprofloxacin was 1 µg L(-1) . These results suggest that hatching, early ontogeny, occurrence of morphological anomalies, antioxidant and biotransformation enzyme activity, and lipid peroxidation in fish can be affected by ciprofloxacin. Environ Toxicol Chem 2016;35:1733-1740. © 2015 SETAC.

Effect of salicylic acid on early life stages of common carp (Cyprinus carpio).

Environmental concentrations of pharmaceutical residues are often low; nevertheless, they are designed to have biological effects at low doses. The aim of this study was to assess the effects of salicylic acid on the growth and development of common carp (Cyprinus carpio) early life stages with respect to antioxidant defence enzymes. An embryo-larval toxicity test lasting 34 days was performed according to OECD guidelines 210 (Fish, Early-life Stage Toxicity Test). The tested concentrations were 0.004, 0.04, 0.4, 4 and 20mg/l of salicylic acid. Hatching, early ontogeny, and both morphometric and condition characteristics were significantly influenced by subchronic exposure to salicylic acid. Also, changes in antioxidant enzyme activity and an increase in lipid peroxidation were observed. The LOEC value was found to be 0.004 mg/l salicylic acid. The results of our study confirm the suggestion that subchronic exposure to salicylic acid at environmental concentrations can have significant effects on aquatic vertebrates.

The effect of the fluoroquinolone norfloxacin on somatic indices and oxidative stress parameters in early stages of common carp (Cyprinus carpio L.).

OBJECTIVES: The aim of this study was to assess the effect of the subchronic exposure of early stages of common carp (Cyprinus carpio L.) to norfloxacin using morphometric data and oxidative stress parameters. METHODS: A subchronic toxicity test was performed on fertilized embryos of common carp according to the OECD Guidelines No. 210. Embryos were exposed to norfloxacin concentrations of 0.0001 (environmental), 0.1, 1.0, 5.0, and 10.0 mg.L(-1) for 34 days. RESULTS: At the end of the test (day 34), significant (p<0.05) stimulation of development was observed in all experimental groups, in contrast to the control. Significantly greater (p<0.01) total body length was also observed in the group exposed to 10.0 mg.L(-1) of norfloxacin compared to the control. A significant increase in the activity of glutathione S-transferase in all carp exposed to norfloxacin concentrations of 0.1 and 1.0 mg.L(-1) (p<0.01), and 5.0 mg.L(-1) (p<0.05) compared to control group was revealed. The activity of glutathione peroxidase was significantly lower (p<0.01) in experimental carp exposed to a norfloxacin concentration of 10.0 mg.L(-1). In experimental carp exposed to a norfloxacin concentration of 0.0001 mg.L(-1), a significant increase (p<0.05) in glutathione reductase activity was found. Significant (p<0.01) decreases in the content of thiobarbituric acid reactive substances in the groups exposed to norfloxacin concentrations of 1.0, 5.0, and 10.0 mg.L(-1) were revealed. CONCLUSION: From the results, we can conclude that norfloxacin has a negative impact on selected biochemical processes related to the production of reactive oxygen species in early-life stages of common carp.

Norfloxacin--toxicity for Zebrafish (Danio rerio) focused on oxidative stress parameters.

The aim of the study was to investigate the effects of subchronic exposure of zebrafish (Danio rerio) to a fluoroquinolone norfloxacin, using selected oxidative stress parameters as a target. Toxicity tests were performed on zebrafish according to the OECD Guidelines number 203 and number 215. In the Subchronic Toxicity Test, a significant (P < 0.01) increase in the activity of glutathione peroxidase, glutathione S-transferase, and catalase was found. In the test, norfloxacin did not affect lipid peroxidation and catalytic activity of glutathione reductase. From the results, we can conclude that norfloxacin has a negative impact on specific biochemical processes connected with the production of reactive oxygen species in fish tested.

Effects of mixture of pharmaceuticals on early life stages of tench (Tinca tinca).

Ubiquitous occurrence of pharmaceuticals in aquatic environment results in concern about potential adverse the effects on nontarget organisms. In water, drugs are present in complex mixtures, in which complicated interactions affect toxicity of single components. The purpose of this study was to examine effect of 35-day-long exposure to mixture of ibuprofen, diclofenac, and carbamazepine on the mortality, growth, early ontogeny, and histopathological changes in tench (Tinca tinca). Early life stage toxicity test was carried out using a modified protocol according to OECD guideline 210. Exposure to mixture of pharmaceuticals at concentration of 60 µg · L(-1) for each substance was associated with significant increase in mortality, as well as significant increase in growth and elevated incidence of malformations. Any of the tested concentrations resulted in histopathological changes of liver, kidney, skin, or gill. After fourteen days of exposure there was short-term delay of development related to increased concentrations of pharmaceuticals in the mixture (2, 20, and 60 µg · L(-1)). Environmentally relevant concentrations (0.02; and 0.2 µg · L(-1)) used in this experiment did not result in toxic impairment of tench.

The effects of subchronic exposure to ciprofloxacin on zebrafish (Danio rerio).

OBJECTIVES: The aim of this study was to assess the impact of ciprofloxacin in Danio rerio on growth, on the development of histopathological changes in selected organs (gills, kidney, liver), and on the activity of some oxidative stress markers during a 28-day toxicity test. METHODS: Juvenile growth tests were performed on D. rerio according to OECD guideline No. 215. Fish at the age of 30 days were exposed for 28 days to a range of concentrations of ciprofloxacin (0.7 µg x L(-1)--environmental concentration, 100, 650, 1100 and 3 000 µg x L(-1)). RESULTS: There were no significant differences between the specific growth rates of fish from the test groups and from the control group. Histopathological examination revealed no pathological changes in organs of fish exposed to any ciprofloxacin concentration. The activity of glutathione S-transferase increased at the ciprofloxacin concentrations of 0.7 and 100 µg x L(-1) compared to the control. A significant decrease in glutathione reductase activity was obtained in fish exposed to ciprofloxacin at 1100 and 3000 µg x L(-1); a significant decrease in glutathione peroxidase activity was also found, but at all tested concentrations except for 100 µg x L(-1). A decrease in the concentration of thiobarbituric acid reactive substances was noted only at 100 µg x L(-1) compared to the control. CONCLUSIONS: According to our results, all tested concentrations of ciprofloxacin, including the environmental concentration, had an influence on oxidative stress markers and detoxifying enzymes in exposed fish, but did not affect fish growth or cause the development of histopathological changes in the fish organism.

The effects of atrazine exposure on early life stages of common carp (Cyprinus carpio).

OBJECTIVES: Atrazine and its chloro-s-triazine metabolites are found in surface water and groundwater of the Czech Republic, although their use has been banned since 2005. The objective of the study was to determine the toxic effects of atrazine at an environmentally relevant concentration of atrazine, 0.3 µg.L-1 and at concentrations of 30, 100, and 300 µg.L-1 on morphometric and condition characteristics, development of early life stages, and antioxidant defense enzymes of common carp. METHODS: The embryo-larval toxicity test was performed according to the OECD Guidelines 210 (Fish, Early-life Stage Toxicity Test). RESULTS: Atrazine exposure showed no effect on morphometric and condition characteristics or histology. Exposure at 0.3 µg.L-1 was associated with significantly increased activity of glutathione peroxidase, glutathione S-transferase, superoxide dismutase, and catalase compared to control. Activity of glutathione reductase was slightly higher at a concentration of 0.3 µg.L-1, with significantly lower (p<0.05) activity observed in groups exposed to 30, 100, and 300 µg.L-1 compared to the group exposed to 0.3 µg.L-1. The level of oxidized lipids was slightly higher in groups exposed to atrazine at 100 and 300 µg.L-1 compared to controls. CONCLUSIONS: Atrazine has a significant influence on the biotransformation enzyme and oxidative defense enzymes of early life stages of common carp. The lowest observed effect concentration (LOEC) was 0.3 µg.L-1.

Evaluation of ibuprofen toxicity for zebrafish (Danio rerio) targeting on selected biomarkers of oxidative stress.

OBJECTIVES: The aim of the study was to investigate the effects of subchronic exposure of zebrafish to ibuprofen, using selected oxidative stress parameters as a target. DESIGN: Toxicity tests were performed on Danio rerio according to OECD No. 203 and No. 215. In the growth test, fish were exposed to subletal concentrations of ibuprofen (0.0001, 0.05, 1, 8, and 25 mg.L-1) for 28 days. For the assessment of free radical defense in fish, the catalytic activities of glutathione reductase (GR), glutathione S-transferase (GST), glutathione peroxidase (GPx), and catalase (CAT), as well as the concentration of malondialdehyde (MDA) were measured. RESULTS: Ibuprofen did not affect the activity of glutathione reductase and catalase. A significant (p<0.01) increase in the activity of glutathione peroxidase was found, which was proved dose-dependent (10.58 nmol NADPH per min per mg protein in the control and 20.53, 26.36, 26.89, and 45.87 nmol NADPH per min per mg protein in the ibuprofen concentrations of 0.5, 1, 8, and 25 mg.L-1. An increased (p<0.05) activity of glutathione S-transferase in the highest concentration was found compared to control. Malondialdehyde levels were found significantly (p<0.01) decreased from control in the concentrations of 0.0001 and 8 mg.L-1, but no dose-dependence was found. CONCLUSION: The results suggest that ibuprofen causes the increase in the activity of some antioxidative and biotransformation enzymes in zebrafish (GPx and GST). We also found a significant decrease in lipid peroxidation in the concentrations of 0.0001 and 8 mg.L-1 compared to control.

Oxidative stress parameters in fish after subchronic exposure to acetylsalicylic acid.

OBJECTIVES: The aim of this study was to investigate the effects of subchronic exposure of juvenile development stages of zebrafish (Danio rerio) to acetylsalicylic acid using selected oxidative stress biomarkers. DESIGN: Toxicity test with acetylsalicylic acid was performed according to the OECD Guideline No. 215, fish D. rerio aged 30 days were used. The tested concentrations were 0.004, 0.4, 40, 120 and 250 mg.L-1, duration of the test was 28 days. Products of lipid peroxidation and antioxidant enzymes were determined as the markers of oxidative stress. RESULTS: Significantly increased glutathione S-transferase activity was found in fish exposed to acetylsalicylic acid concentrations 40, 120 and 250 mg.L-1. The highest values of glutathione reductase activity were found in the groups exposed to acetylsalicylic acid concentrations 0.4, 40 and 120 mg.L-1. In the group exposed to acetylsalicylic acid concentrations 40 mg.L-1, catalase activity was significantly higher compared to the control group. Significantly higher glutathione peroxidase activity was found in the groups exposed to acetylsalicylic acid concentrations 0.004 and 120 mg.L-1. The concentrations of TBARS were lower in fish exposed to acetylsalicylic acid at all tested concentrations compared to control. CONCLUSION: The subchronic exposure of zebrafish to acetylsalicylic acid causes an increase in activity of antioxidant and biotransformation enzymes and a decrease in lipid peroxidation.

Acute toxicity of acetylsalicylic acid to juvenile and embryonic stages of Danio rerio.

OBJECTIVES: The aim of this study was to compare the acute toxicity of acetylsalicylic acid to embryonic and juvenile stages of aquarium fish - zebrafish (Danio rerio), oxidative stress parameters and detoxifying enzyme. METHODS: Tests were performed according to OECD No. 203 (Fish, acute toxicity test) and OECD No. 212 (Fish, short-term toxicity test on embryo and sac-fry stages) methodology. RESULTS: The results showed the mean acetylsalicylic acid LC50 value to be 567.7 mg/L in juvenile zebrafish. The acute toxicity of acetylsalicylic acid for zebrafish embryos was 274.6 mg/L. Statistically significantly higher activity of GST was found in concentrations 340, 380 and 420 mg/L of acetylsalicylic acid. TBARS, GPx and GST didn't show statistically significant activity in tested concentrations of acetylsalicylic acid. CONCLUSIONS: The results revealed a statistically significantly higher degree of sensitivity in the embryonic stages of zebrafish compared to its juveniles. Acetylsalicylic acid did not cause statistically significantly higher antioxidative defence in zebrafish.