Hematological changes, redox imbalance, and changes in Na+/K+-ATPase activity caused by bisphenol-A and the integrated biomarker responses in Labeo rohita (Hamilton, 1822).

Bisphenol-A (BPA) is a plasticizer commonly used in the plastics industry to manufacture plastic materials. It is abundant in aquatic ecosystems, resulting in increased contamination and lower concentrations that may represent a significant threat to the aquatic system. Hence in the present study, an Indian major carp, Labeo rohita, was exposed to two different BPA concentrations (1 and 10 µg/L) for 30 days. Compared to control, the chronic effects resulted in significant alterations in red blood cell (RBC) and white blood cells (WBC) count. The exposure to BPA caused significant changes in antioxidant activity in gill, liver, and kidney tissues (inferred by catalase, glutathione peroxidase, and glutathione S-transferase activity) in L. rohita. Regarding lipid peroxidation (LPO), we observed an increase in liver and kidney alteration, while LPO was noted in gill tissue compared to the control. Furthermore, increased Na+/K+-ATPase activity was observed in gills at the end of the 10th day and a gradual decrease at the end of the 30th day. These results indicated that exposure to BPA alters the RBC and WBC levels, antioxidant enzyme activity (gills, liver, and kidney), and Na+/K+-ATPase activity in the gill of L. rohita exposed to BPA (at 1 and 10 µg/L). Therefore, our findings will help us gain better insight into the toxicity of BPA in freshwater ichthyofauna.

Evaluation of antioxidant response and Na+-K+-ATPase activity in zebrafish exposed to polyethylene microplastics: Shedding light on a physiological adaptation.

Although the toxicity of microplastics has already been demonstrated in different animal models, our knowledge about the response of freshwater fish to this pollutant is still limited. Thus, we aimed to evaluate the impact of exposure of zebrafish adults (Danio rerio) to environmentally relevant concentrations of polyethylene microplastics (PE-MPs) (5 and 50 µg/L) and at different times of exposure (10 and 20 days). Initially, scanning electron microscope image illustrated size and format of the particle and FTIR analysis confirmed the presence of PE-MPs in the gastrointestinal tract of fish (at both concentrations tested). Subsequently, an alteration of oxidative and antioxidant responses was evaluated in the liver and brain. The results showed that catalase (CAT) activity, in liver, was significantly decreased, as was glutathione S-transferases (GSTs) activity (on the 10th experimental day). However, after 20 days of exposure, we observed a concentration-dependent increase in GST activity in liver of the animals exposed to PE-MPs. Furthermore, the lipid peroxidation (LPO) level was significantly increased by exposure to MPs, especially in the brain, after 20 days of exposure. The increase in Na+-K+-ATPase activity in the animals' gills was correlated with the increased production of malondialdehyde (MDA), which suggests the existence of a compensatory mechanism in which the high activity of this enzyme would be necessary to regulate the loss of ions caused by the increase in the processes of LPO, which has never been previously demonstrated. Thus, our study sheds light on a new physiological adaptation to deal with the oxidative effects of PE-MPs, in addition to supporting the future use of the assessment of Na+/K+-ATPase activity as a biomarker of the toxicity of these pollutants.

Transcriptional, biochemical and histological alterations in adult zebrafish (Danio rerio) exposed to benzotriazole ultraviolet stabilizer-328.

The occurrence of Benzotriazole Ultraviolet Stabilizer-328 (BUV-328) in different environmental and biological matrices is of immediate environmental concern. In the present study, we evaluated the toxicity of BUV-328 in zebrafish liver tissues to understand the role of oxidative damage in hepatotoxicity. Adult zebrafish were exposed to 0.01, 0.1 and 1 mg/L of BUV-328. At the end of 14, 28 and 42 days, liver tissues were examined for the responses of antioxidant enzymes, gene expression and histopathological alterations. The results indicated that superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities were elevated at concentrations of 0.1 and 1 mg/L on 14th and 28th day. Glutathione S-transferase (GST) activity and malondialdehyde (MDA) levels were elevated in all the treated groups. The transcriptional levels of genes encoding sod, cat, gpx and gst enzymes were increased at 14th day and then declined (except sod on 28th day). Moreover, transcription of cyp1a and hsp70 were up-regulated throughout the study period. Histopathological lesions such as hypertrophy, cellular and nuclear enlargement, cytoplasmic and nuclear degeneration, necrosis with pyknotic nuclei, lipid and cytoplasmic vacuolization and nuclear displacement to the periphery were found to be increased with the dose and exposure duration. In brief, our findings indicate that even a low dose of BUV-328 is toxic to induce oxidative stress and liver damage in zebrafish over a long period of exposure.

Comparative toxicity of UV-filter Octyl methoxycinnamate and its photoproducts on zebrafish development.

In the present study, we explored the adverse effects of Octyl methoxycinnamate (OMC), and its photoproducts, namely 2-ethylhexanol (2-EH) and 4-methoxybenzaldehyde (4-MBA) on the developmental stages of zebrafish using various biomarkers such as developmental toxicity, oxidative stress, antioxidant response, neurotoxicity and histopathological changes. The 96 h effective concentrations (EC50) of OMC, 2-EH and 4-MBA were found to be 64.0, 34.0 and 3.5 µg/mL, respectively in the embryo toxicity test. Embryos exposed to the EC50 of OMC, 2-EH and 4-MBA showed time-dependent increases in the malformation, heart rate and hatching delay. The lipid peroxidation (LPO) level was significantly (p < 0.05) increased and both induction and inhibition of SOD, CAT, GPx and GST activities were observed in the zebrafish embryos exposed to OMC, 2-EH and 4-MBA. GSH activity was significantly (p < 0.05) decreased in the highest exposure groups, when compared with the control. AChE activity was increased in lower concentrations of OMC, 2-EH and 4-MBA exposed embryos whereas, the activity was found to be decreased in highest concentration. Moreover, the histopathological studies showed severe damage to the muscle fibers and yolk sac regions of the larvae with 4-MBA treatment. The photoproduct 4-MBA has the highest toxic effect, followed by 2-EH and OMC. Our results provide useful insights into the impacts of OMC and its photoproducts on zebrafish development.

DNA damage and physiological responses in an Indian major carp Labeo rohita exposed to an antimicrobial agent triclosan.

This study is aimed to evaluate the toxic effects of triclosan (TCS) in an Indian major carp Labeo rohita. The 96-h LC50 value of triclosan to L. rohita was found to be 0.39 mg L-1. Fish were exposed to two sublethal concentrations (0.039 mg L-1, treatment I and 0.078 mg L-1, treatment II) of TCS for 35 days, and certain hematobiochemical, antioxidant, histopathological responses were measured. Compared to the control group, there was a significant (p < 0.05) decrease in the values and genotoxicity of hematological parameters such as hemoglobin (Hb), hematocrit (Hct), and erythrocyte (RBC) in TCS-exposed fish, but the values of leucocyte count (WBC), mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH) were found to be increased. A biphasic response in mean corpuscular hemoglobin concentration (MCHC) value was observed during the study period (35 days). Significant (p < 0.05) alterations in plasma biochemical parameters (glucose and protein), electrolytes (Na+, K+, and Cl-), and transaminases (GOT and GPT) were observed in fish treated with TCS in both treatments. Gill Na+/K+-ATPase activity was found to be decreased in fish treated with TCS in both treatments. Enzymatic and nonenzymatic antioxidant index levels have also fluctuated in all the tissues (gill, liver, and kidney). The histological lesions were comparatively more severe in the gill than the liver and kidney. Comet assay showed DNA damage on exposure at two sublethal concentrations. The present results suggest that TCS is highly toxic to fish even at sublethal concentrations.

Developmental toxicity and biological responses of zebrafish (Danio rerio) exposed to anti-inflammatory drug ketoprofen.

Ketoprofen a nonsteroidal anti-inflammatory drug (NSAID) is widely used in over-the-counter to treat pain, swelling and inflammation. Due to extensive application these drugs has been detected in surface waters which may create a risk to aquatic organisms. The aim of the present study is to assess the ecotoxicity of ketoprofen at different concentrations (1, 10 and 100 µg/ml) on embryos and adult zebrafish (1, 10 and 100 µg L-1) under laboratory conditions. In embryos, concentration dependent developmental changes such as edema, spinal curvature, slow heartbeat, delayed hatching, and mortality rate were observed. In adult zebrafish, biochemical enzymes such as AST, ALT and LDH activities were significantly (P < 0.05) increased whereas a decrease in Na+/K+-ATPase activity was noticed in all the tested concentrations of the drug ketoprofen. Similarly, exposure of ketoprofen caused a significant decrease in antioxidant levels in liver tissue (SOD, CAT, GSH, GPx, and GST). However, lipid peroxidation (LPO) level in liver tissue was found to be increased. The histopathological studies further evidenced the impact of ketoprofen in the liver tissue of zebrafish. The present result concludes that ketoprofen could have an impact on the development and biological endpoints of the zebra fish at above concentrations. The malformation in the development of the embryo and changes in the biological end points may provide integrated evaluation of the toxic effect of ketoprofen on zebrafish in a new perspective.

Green synthesis of silver nanoparticles using Piper nigrum: tissue-specific bioaccumulation, histopathology, and oxidative stress responses in Indian major carp Labeo rohita.

The aim of the present investigation is to assess the sublethal toxicity of biologically synthesized silver nanoparticles (Ag NPs) in Indian major carp Labeo rohita. Ag NPs used in the study were synthesized by using AgNO3 with aqueous leaf extract of Piper nigrum. Median lethal concentration (LC50) of synthesized Ag NPs was determined for 96 h (25 µg/L); 2.5 µg/L (1/10th LC50) and 5 µg/L (1/5th LC50) were taken as sublethal concentrations to evaluate the toxicity for 35 days. The results of the TEM, SEM, and EDX analyses revealed that Ag NPs were considerably accumulated in the gill, liver, and kidney of fish at both concentrations (2.5 and 5 µg/L). Consequently, the activity of the antioxidant enzymes, SOD and CAT, was significantly (P < 0.05) decreased in the gill, liver, and kidney when compared to the control group during the study period. However, lipid peroxidase (LPO) activity in the gill, liver, and kidney was significantly (P < 0.05) increased, and the result concluded a possible sign of free radical-induced oxidative stress in Ag NP-exposed fish than the sham-exposed individuals. The histopathological study also confirmed the alterations such as degeneration of lamella, lifting of lamellar epithelium, hepatic necrosis, pyknotic nuclei, increased intracellular space, and shrinkage of glomerulus elicited by Ag NPs in the gill, liver, and kidney of Labeo rohita with two different concentrations. The findings of the present study revealed that green synthesis of Ag NPs from Piper nigrum at sublethal concentrations leads to accumulation of Ag, oxidative stress, and histopathological alterations in the target organs of the fish, Labeo rohita.

Sublethal toxicity of quinalphos on oxidative stress and antioxidant responses in a freshwater fish Cyprinus carpio.

Extensive use of quinalphos, an organophosphorus pesticide, is likely to reach the aquatic environment and thereby posing a health concern for aquatic organisms. Oxidative stress and antioxidant responses may be good indicators of pesticide contamination in aquatic organisms. The data on quinalphos induced oxidative stress and antioxidant responses in carps are scanty. This study is aimed to assess the two sublethal concentrations of quinalphos (1.09 and 2.18 µL L-1 ) on oxidative stress and antioxidant responses of Cyprinus carpio for a period of 20 days. In liver, the malondialdehyde level was found to be significantly increased in both the concentrations. The results of the antioxidant parameters obtained show a significant increase in superoxide dismutase, catalase, and glutathione-S-transferase activity in liver of fish. These results demonstrate that environmentally relevant levels of the insecticide quinalphos can cause oxidative damage and increase the antioxidant scavenging capacity in C. carpio. This may reflect the potential role of these parameters as useful biomarkers for the assessment of pesticide contamination. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1399-1406, 2016.