Acute toxicity, biochemical and histopathological responses of endosulfan in Chanos chanos

https://doi.org/10.1016/j.ecoenv.2016.05.013Get rights and content

Highlights

  • The LC50 of endosulfan in Chanos chanos juvenile was found to be 21.5 g/L at 96 h.

  • Acute endosulfan exposure causes generation of ROS and oxidative stress in C. chanos at 96 h.

  • Acute exposure to endosulfan in C. chanos cause neurosupressive and altered metabolic enzymes.

  • Endosulfan exposure induced histopathological changes in C. chanos at 96 h.

Abstract

This study investigated 96 h median lethal concentration of endosulfan (99%, pure α: β ratio of 7:3) by conducting static non-renewable acute toxicity bio-assay in Chanos chanos juvenile with average weight (110±5.65 g). Further, the effect of different definitive doses (18.5, 19.5, 20.5, 21.5 and 22.5 µg/L) of endosulfan on metabolic, heamato-immunoligcal and histopathological response were probed. Anti-oxidative enzymes CAT, SOD and GST showed significant (p<0.01) increase of activity in the liver, gill and brain during exposure to endosulfan in a dose and time dependent manner. The brain AChE activity showed significant (p<0.01) inhibition from 18.5 to 22.5 µg/L exposure of endosulfan than the control group. LDH and MDH activity gradually increased with consequent increasing dose of endosulfan exposure in the liver, gill and brain. Similarly, ALT, AST and G6PDH activities in both liver and gill increased with consequent increases in the dose of endosulfan exposure. Immunological profile such as blood glucose and serum cortisol level significantly enhanced while respiratory burst activity declined with consequent increasing doses of endosulfan exposure. Histopathological alteration in the gill demonstrated curling of secondary lamellae, thickening of primary epithelium, shorting of secondary lamellae, epithelial hyperplasia, fusion of secondary lamellae, aneurism, and collapsed secondary lamellae due to dose dependent exposure of endosulfan. Liver histology illustrated cloudy swelling and necrosis with pyknotic nuclei to the moderate dose of endosulfan, whereas higher dose of endosulfan (21.5 µg/L) displayed severe necrosis of hepatic cells. Overall results clearly indicate that acute exposure of endosulfan led to pronounced deleterious alterations on biochemical, heamato-immunological, and histopathological responses of C. chanos juvenile.

Introduction

Agriculture effluents contain highly toxic chemicals viz. pesticides, which can causes a hazardous effect to the aquatic environment. The accumulation and persistence of pesticides in the aquatic environment found a threat to biological life, as seen by the acute and chronic poisoning of aquatic organisms (Hemmer et al., 2001, Wirth et al., 2001). Organochlorines are utmost hazardous, since they are very persistent, non-biodegradable and residue accumulate in food chain (Jaffery et al., 1990).

Endosulfan (6,7,8,9,10,10- hexachloro 1, 5, 5a, 6, 9, 9a-hexahydro- 6, 9-methano-2, 3, 4-benzodioxathiepine-3-oxide, CAS No. 115-29- 7) is an organochlorine pesticide, still in vogue in many parts of the world for combating pests in order to increase agricultural productivity (Kullman and Matsumura, 1996). Due to its indiscriminate usage and potential transport from the point of its original application, the residues of have been detected in the atmosphere, soils, sediments, surface, rain waters, and foodstuffs (Miles and Pfeuffer, 1997). Residues of endosulfan or its metabolites have been found to be associated with the mortality of fish in continental aquatic systems (Dalela et al., 1979; Ferrando, 1989; Gimeno et al., 1995) and further degrade with major end product as endosulfan sulfate. This degraded product is even more persistent and toxic to many aquatic species than the parent compound (Shimmel et al., 1977). The concentrations of organochlorines in the sea around India have been reported to be high compared to other regions (Tanabe and Tatsukawa, 1980). Large amount of endosulfan residue has been reported in Indian estuaries (Bhattacharya et al., 2003).

The exposure to contaminants in aquatic ecosystems can enhance the intracellular formation of reactive oxygen species (ROS), which induce oxidative damage to biological systems (Giulio et al., 1989). Different classes of pesticides may induce in vitro and in vivo generation of ROS, such as hydrogen peroxide, superoxide and the hydroxyl radical (Bagchi et al., 1995). Oxidative stress arises when an imbalance occurs between production and elimination of ROS. The enzyme defense system can detoxify ROS, comprising superoxide dismutase (SOD), catalase (CAT), while organic peroxides can be detoxified by the activity of glutathione-S-transferase (Halliwell and Gutteridge, 1989).

The biochemical parameters are very sensitive to acute concentrations of many stress agents such as organochlorine. Therefore, it is preferable to observe the alterations of general stress biomarker parameters (LDH, MDH, ALT, AST and G6PDH) to determine stress situation in an organism (Kumar et al., 2014a, Kumar et al., 2016a).

Haemato-immunological parameters acts as indicators for the assessment of the physiological status of the organisms exposed to abiotic and biotic stressors. Change in cortisol acts as excellent primary stress indicator of functional alterations in the hypothalamo-pituitary-interrenal (HPA) axis (Hontela, 2005). Variation in blood glucose is the most important indicator of secondary stress response (Kumar et al., 2011a, Kumar et al., 2016b), whereas enhanced respiratory burst activity due to stress represents the increased bacterial killing power of phagocytes leading to improved immunity in fishes (Kumar et al., 2014b, Kumar et al., 2014c; Gupta et al.,, Gupta et al., 2014b).

AChE is one of the most commonly used enzyme, recognized as a biomarker for environment pollution studies in fishes. In general, fishes can endure about 70–80% inhibition of AChE activity in cerebellum, which controls the muscular co-ordination (Kumar et al., 2011b, 2012; Gupta et al., 2014b).

Histopathology illustrations acts as a powerful and quick tool to detect effects of environmental pollutants (Johnson et al., 1993). Histopathological investigations demonstrate prominent alterations in the gills (Poleksic and Karan, 1999) and liver (ICES, 1997) tissues in order to determine the effect of pollution. Gills are extremely important and most sensitive because of the intimate and continuous contact with the water, whereas as the liver is prime organ for the detoxification of xenobiotic.

Milk fish (Chanos chanos) is widely distributed and cultured throughout the tropical and subtropical Indo-Pacific region (Swanson, 1996). The present production of C. chanos estimated to be 943, 259 t (FAO, 2012). The Milk fish has been the subject of toxicity studies due to its consumer preferences and commercial demand in the Asian market (Kumar et al., 2014a, Kumar et al., 2016a, Kumar et al., 2016b, Magesh and Kumaraguru, 2006).

To the best of our understanding, acute toxicity, heamato-biochemical and histopathological responses of endosulfan in Chanos chanos has not been studied therefore, present study was aimed to determine LC50 of endosulfan and consequently to assess the effect of 96 h exposure of different doses of endosulfan on metabolic, heamato-immunoligcal and histopathological response of C. chanos.

Section snippets

Experimental animals and condition

Juvenile milkfish (Mean weight ± standard error, 110±5.65 g) were acquired from the Central Institute of Brackishwater Aquaculture (CIBA), Muttukudu Research Centre, Chennai, India and were acclimated in fiber tank to experimental conditions for 15 days before starting the experiment. The aeration was provided round the clock and water temperature recorded to be in the range of 26.0–28.6 °C and 35 ppt salinity (Supplement Table 1). The animals were fed with practical diet (30% crude protein) at a

Median lethal concentration

The acute toxicity of endosulfan and cumulative mortality of C. chanos at 24, 48, 72 and 96 h and corresponding LC50 values with 95% confidence limits are presented in Table 1. The LC50 values during 24, 48, 72 and 96 h of endosulfan exposure were found to be 25.3, 23, 22.2 and 21.5 µg/L, respectively and there was significant difference observed between 24 and 96 h LC50 values.

Antioxidative enzymes

The activities of antioxidative enzymes CAT, SOD and GST in liver, gill and brain of C. chanos are presented in Table 2.

Median lethal concentration

The LC50 procedure is easy and most accepted for acute toxicity study; moreover it is based on 50% mortality of the experimental animals in a fixed duration (96 h). The acute toxicity studies is based on mortality due to easy determination and also has biological and ecological importance. Various researchers have reported LC50 of endosulfan in fishes (Schimmel et al., 1977, Schimmel et al., 1983, Magesh and Kumaraguru, 2006, Hemmer et al., 1992, Hemmer et al., 2001) and is Supplement Table 2.

Conclusion

Endosulfan is a highly toxic synthetic pesticide widely used in agriculture and finally reaches to river through runoff and thus affecting physiological and biochemical status of the non target aquatic organism. Overall results of this study indicate that acute exposure of endosulfan led to pronounced deleterious effect on biochemical heamato-immunological parameters, and histopathology of C. chanos. Therefore, such data are useful in ecological risk assessment, but with certain limitations and

Acknowledgement

The authors are grateful to Dr. P. S. Minhas, Director, ICAR-National Institute of Abiotic Stress Management, Baramati, Pune and Director, ICAR-Central Institute of Brackish Water Aquaculture, Chennai, India for providing support for the study. We also thank to Mr. D. Srinivasan for the technical assistance rendered by him during the experimental trial.

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