Oxidative stress biomarkers, biochemical responses and Na+ -K+ -ATPase activities in Nile tilapia, Oreochromis niloticus exposed to diclofenac

https://doi.org/10.1016/j.cbpc.2020.108934Get rights and content

Highlights

  • Exposure to diclofenac affected the antioxidant defence system in Nile tilapia.

  • Sub lethal diclofenac altered serum biochemical parameters in fish.

  • Sub lethal diclofenac induced lipid peroxidation in O. niloticus.

  • Diclofenac inhibited Na+ -K+ -ATPase activities in O. niloticus.

Abstract

The residues and metabolites from pharmaceuticals have been noted to cause adverse effects to both target and non-target aquatic organisms. The sublethal effects of diclofenac at 0.17, 0.34 and 0.68 mg L−1 on oxidative stress biomarkers, biochemical responses and Na+ -K+ -ATPase activities in the gill tissue of Nile tilapia, Oreochromis niloticus were investigated for 60 days. Elevated levels of some serum biochemical parameters including protein, glutamic oxalacetic transaminase, glucose, glutamic pyruvic transaminase, lactate dehydrogenase, alkaline phosphatase and also some catalysts of gluconeogenic enzymes such as glucose-6-phosphatase, fructose 1, 6 bisphosphatase in the fish liver, increase as the concentration of the diclofenac increased. The reactions of glutathione-S-transferase, catalase, lipid peroxidation, superoxide dismutase, glutathione peroxidase, carbonyl protein and reduced glutathione were elevated (p < 0.05) while the activities of Na+ -K+ -ATPase was significantly reduced (p < 0.05) in fish gill, indicating an adaptive response strategies to mitigate the impact of the drug on the exposed fish. Chronic exposure to sublethal diclofenac can induce oxidative stress and modulates serum biochemical indexes of O. niloticus, suggesting the need for close monitoring of the drug and their metabolites in aquatic environment considering the possible potential adverse effects it may cause even to non-target organisms.

Introduction

Drugs and its metabolites have been reported to found its way through erosion and other anthropogenic sources into the aquatic environment where they accumulate and cause deleterious effects to the inhabitant aquatic organisms. The presence of residues of these drugs and their metabolites in aquatic ecosystem have attracted the attention of many scientists for the need to find the solution to avert prolonged effects these pharmaceuticals can cause to aquatic organisms.

Diclofenac, is a nonsteroidal anti-inflammatory group of drugs which is applied to relieve pain both in human and livestock. Diclofenac is implicated as an emerging contaminant because of its persistence in aquatic environment as well as its hydrophilicity and stability (Madikizela et al., 2018; Tiedeken et al., 2017).

Diclofenac was found to accumulate in edible fruits and vegetables where it could directly cause human health implications (Bartrons and Peñuelas, 2017; González García et al., 2018). Also increase in usage of pharmaceuticals to treat illness, emptying of wastewaters which are not treated into the environment, may increase the availability of drugs including diclofenac in the water body (Fekadu et al., 2019; Williams et al., 2019).

Exposure to environmental concentration of diclofenac altered stability of lysosomal membrane and cyclooxygenase (COX) reactions as well as DNA damage in Perna perna (Fontes et al., 2018). Environmental concentration of diclofenac at 2.5 ngL−1 caused genotoxicity and DNA alteration in Mytilus galloprovincialis (Mezzelani et al., 2018). Chen et al. (2015) found that survival and reproduction potentials of Folsomia candida were hampered by diclofenac toxicity in soil. Chae et al. (2015) demonstrated that diclofenac had definitive and toxic effects on developmental stages of Xenopus embryos. Through biomagnification in the food chain, diclofenac could also illicit potential ecological consequences to both target and non-target organisms (Proffitt and Bagla, 2004).

Na+ -K+ -ATPase activities are known to be inhibited in aquatic organisms due to xenobiotics by hampering metabolic pathways that produce energy (Haya and Waiwood, 1983). The activity of the enzyme can be used to evaluate the adverse effects of pollutant on the physiological integrity of the exposed organisms (Agrahari and Gopal, 2008). The level of serum glutamic oxalacetic transaminase and serum glutamic pyruvic transaminase in fish blood as well as the activities of metabolic enzymes including lactate dehydrogenase, alkaline phosphatase, glucose-6-phosphatase, fructose 1, 6 bisphosphatase in fish liver could be important biomarkers to evaluate cellular dysfunction due to contaminants in fish (Malarvizhi et al., 2012).

Under stressful conditions, due to toxic compounds, free radical and reactive oxygen species (ROS) can be produced which impairs the physiological parameter of fish. Consequently, some antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase glutathione reductase and reduced glutathione (a non-enzymatic antioxidant) are produced by fish in order to neutralise and protect itself against ROS.

The effect of diclofenac on the serum biochemistry, oxidative stress enzymes and Na+ -K+ -ATPase activity in O. niloticus are not reported extensively. Nile tilapia is used in this investigation because of its global distribution in freshwater ecosystem and can withstand variation in environmental condition in laboratory. The study was designed to investigate if chronic exposure to diclofenac can alter the antioxidant defence system, serum biochemistry and Na+ -K+ -ATPase activities of Nile tilapia, O. niloticus.

Section snippets

Fish and drug

O. niloticus of mean weight 40.48 ± 2.15 g and mean length 12.98 ± 1.01 cm; numbering one hundred and thirty (130) procured from Nalasopara, India were used for the investigation. Acclimatization of the fish was done in 500 L capacity tank for two weeks. The fish were fed with pelleted feed twice daily. Fecal droppings and remnants of unconsumed feed were removed regularly to prevent fouling the experimental media. Diclofenac (purity, 99%, Danish Laboratory, India) was purchased from

Biochemical responses

Levels of both total serum protein and glucose as presented in Fig. 1, show that sublethal exposure to long term diclofenac significantly inhibited (p < 0.05) the level of protein and elevated the glucose level after the 60 days of experimental period at high concentration of the diclofenac. SGOT/AST and SGPT/ALT levels significantly (p < 0.05) increased at the end of the study in comparison to the control. The activities of the enzymes increased at the high concentration of the drug. The

Discussion

The study has shown that chronic treatment with sublethal diclofenac can alter the activities of antioxidant and Na+–K+-ATPase enzymes in the gill, the liver function enzymes as well as the metabolic enzyme activity in Nile tilapia in a duration and dose dependent association.

Exposure to sub lethal diclofenac reduced the serum protein level and increased significantly the glucose level as well as the serum AST and ALT in the liver. The reduction in protein could be a result of the adverse

Conclusion

The present investigation has shown that sublethal diclofenac can affect the serum biochemistry, cause oxidative stress and also inhibit Na+/K+ -ATPase reaction of the treated fish in a duration and dose dependent relationship. The results of the investigation indicate that the observed changes found in the exposed organisms compared with the control could be adaptive responses mechanism developed by the fish to counteract the adverse impact triggered by the drug on the vital organs of the

Ethical approval

All applicable international, national and institutional guidelines for the care and use of animals were followed (The research undertaken complies with the current animal welfare laws in India. The study was approved by the Board of Studies of Fish of the Central Institute of Fisheries Education (Deemed University), Mumbai, India. The care and treatment of animals used in this study were in accordance with the guidelines of the CPCSEA (Committee for the Purpose of Control and Supervision of

Declaration of competing interest

I, Malachy Ajima declare that I have to conflict of interest. Pramod K. Pandey declares that he has no conflict of interest; Kundan Kumar declares that he has to conflict of interest; Nalini Poojary declares that she has no conflict of interest.

Acknowledgements

The authors wish to thank the Indian – African Fellowship programme for the doctoral fellowship granted to Dr. M.N.O. Ajima. We also wish to thank the Director, ICAR-Central Institute of Fisheries Education, Mumbai, India for providing the equipment and facilities, used for the study.

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