Blood Biochemical and Erythrocytic Morpho-pathological Consequences of Naphthalene Intoxication in Indian Teleost, Anabas testudineus (Bloch)

https://doi.org/10.1016/j.etap.2020.103490Get rights and content

Highlights

  • Two doses of sub-lethal concentration of naphthalene applied on A. testudineus

  • Responses analysed based on doses and exposure period

  • Investigation on blood biochemical parameters

  • Morpho-pathological analysis of erythrocytes

Abstract

Anabas testudineus (Bloch) was exposed to 0.71 mg/L and 1.42 mg/L (25 and 50% of LC50 value respectively) naphthalene, a polycyclic aromatic hydrocarbon (PAH), for 21 days. Blood biochemical parameters and erythrocytic morphological alterations were assessed to describe the naphthalene toxicity. Biochemical analysis showed a significant increase in glutamic pyruvic transaminase, GPT (576.7 ± 11.79 and 608.9 ± 12.08 U/L, respectively) and alkaline phosphatase, ALP (12.9 ± 0.69 and 13.4 ± 0.64 U/L, respectively) activities under two doses compared with control. Protein and albumin (ALB) content in blood decreased significantly, in comparison with control value in the tune of 22.67 ± 1.04 and 23.97 ± 1.24 g/dl, respectively and 10.7 ± 0.79 and 11.1 ± 0.67 g/dl, respectively. Erythrocytes showed varied symptomatic morphological changes under naphthalene exposure, which included severe denaturation, swelling in cells, appearance of sickle and tear drop cells, and cellular vacuolation. In particularly, the changes were more prominent under higher naphthalene exposure. Following the results, it has been able to establish that GPT, ALP, protein and ALB, and the morphological manifestations of erythrocytes would be good tools of biomarker in monitoring toxicological paradigm, especially to naphthalene exposure in aquatic bodies.

Introduction

Worldwide massive and irregular extraction and exploration of natural resources from the earth, unplanned anthropogenic forest fires, uncontrolled oil-spills, etc., invite subconscious natural environmental degradation causing a serious conflict between man and environment (Bautista et al., 2019). Accordingly, contamination of aquatic resources has been recognized as a concern for augmenting vital awareness for ‘save water quality’ programme worldwide (Petersen et al., 2017; Bautista et al., 2019). Naphthalene addressed here is a polycyclic aromatic hydrocarbon (PAHs) that are widely distributed in soil, water, air and aquatic environments (Slezakova et al., 2013; Nakata et al., 2014). It is an important constituents of petroleum fuels (Honda and Suzuki, 2020). Abundance and ubiquitous distribution of naphthalene due to different man-made activities, such as oil-spillage, deposition in different media, terrestrial discharge and runoff, effluents from domestic and industrial sources, etc., recognized as one of the important contributors of water pollution (Alderman et al., 2020). Aquatic organisms suffering from different type of lesions, viz., tissue damage, cellular lesions, ulceration, and necrosis, etc., mainly due to presence of lipophilic xenobiotics mainly PAHs in aquatic environment (Kennedy, 2014; Madison et al., 2017; Alsaadi et al., 2018). Generally, the impact of these xenobiotics is concerned with specific toxicity (Philibert et al., 2016; Honda and Suzuki, 2020). However, the toxic outcomes are evaluated based on their structural and functional damage intensity/potentiality within the biological system (Agamy, 2013; Medeiros et al., 2017; Alderman et al., 2020). PAHs undergo subsequent bioaccumulation in bottom dwellers, including mollusks and fish species (Barhoumi et al. 2016) due to their non-degradability and persistent nature in the medium. In case of filter feeders and fish, intake of food directly through gill (Cheikyula et al., 2008) is the easy entrance which leads to disturbance in the vital physiological functions (Elumalai and Balasubramanian, 1999) that ultimately affecting the total aquaculture production. In addition, PAHs and their metabolites are recognized as carcinogenic and mutagenic (Katsumiti et al., 2009; Stimmelmayr et al., 2018) and it also revealed that their toxicosis is concerned largely with the extent of exposure (Silva et al., 2006). The PAHs concentration in different aquatic media induces several health risk especially to fish (Kennedy, 2014; Alsaadi et al., 2018). Generally, the naphthalene concentration in natural water environment was detected in the range between 0.1 and 10 ng/L (Ahmad et al., 2003), in fish concentration ranges from 0.03 to 1.004 μg/g (Hossain et al., 2014). Till today, the majority of available data on naphthalene toxicity in freshwater organisms i.e., fish are mainly on mortality, life history trait responses, histological alterations, endocrine disruption, iono-osmoregulatory disruption and immunotoxicity (Elumalai and Balasubramanian, 1999; US EPA, 2003; Santos et al., 2006; Hossain et al., 2014). Recently, genotoxic nature of naphthalene in fish species has been demonstrated by several authors (Pacheco and Santos, 2001; Mekkawy et al., 2011). Apart from these, toxicological effects of naphtalene on other aquatic biota such as crab (Scylla serrata), prawn (Metapenaeus affinis and Penaeus semisulcatus), Walleye (Sander vitreus), Herring Gull (Larus argentatus) and even on algae (Chlorella sp.) have been documented by several authors (Vijayavel and Balasubramanian, 2006; Kong et al., 2010; McGoldrick et al., 2018; Soltani et al., 2019). However, limited information on naphthalene toxicity exists currently on blood biochemistry and erythrocytes morpho-pathological alterations and the utility of extrapolating data regarding this on naphthalene intoxication is unknown. Additionally, the linkage among blood biochemical parameters and erythrocytes morpho-pathological alterations has limited information. Recently we have demonstrated the importance of linking blood biochemical parameters and erythrocytes’ morpho-pathological alterations under anthracene exposure in Anabas testudineus (Dey and Ghosh, 2019). These data addressed the necessity for linking blood biochemical parameters and erythrocytes’ morpho-pathological responses under naphthalene toxicosis to understand the pollution status using aquatic organism.

Usually, haematological parameters are evidenced as indicators of general health status of animals including fish (Kreutz et al., 2011; García et al., 2015). Blood biochemistry and erythrocytes pathology attributed as convenient and reliable diagnostic tool (Sepici-Dinçel et al., 2009; Kavitha et al., 2010; Bojarski and Witeska, 2020) to assess toxic outcomes of a large number of xenobiotics (Ronda et al., 2018; Bautista et al., 2019; Lin et al., 2019). Wepener (1997) established the toxicosis of PAHs through evaluation of growth parameters, oxygen intake rate of fish and some bioenzymological parameters. Few workers linked erythrocytic alterations and its morphology with genotoxic property of xenobiotics mainly PAHs in fish species (Bombail et al., 2001; Bushra et al., 2002). Recently, Talapatra and Banerjee (2007); Mekkawy et al. (2011) and Sayed and Authman (2018) addressed unusual nuclear abnormalities and morphological changes in erythrocytes as cytotoxic and genotoxic indicators in fish. However, few studies linked altered environmental quality with damaged haematopoietic tissues and blood biochemistry due to naphthalene intoxication (Pacheco and Santos, 2001; Mekkawy et al., 2011).

In this study, therefore, naphthalene (PAH) induced alterations on blood biochemistry and erythrocyte morpho-pathology in air-breathing carnivorous teleostean fish, Anabas testudineus (Bloch) has been investigated. In this study, we have selected Anabas testudineus as model species because of its wide availability and distribution in freshwater ecosystem, easy adaptability under laboratory condition and commercial importance as high rich protein source (Samanta et al., 2018). Apart from these our previous studies have been demonstrated its capability as early warning signal of xenobiotic exposure under both laboratory and field conditions (Samanta et al., 2017, 2018).

Section snippets

Procurement and maintenance of fish species

Air-breathing freshwater carnivorous teleost, Anabas testudineus (Bloch) having average weight and total length of 32.19 ± 1.21 g and 11.48 ± 0.924 cm, respectively brought from registered fish farm. Acclimation was done for fortnight in large-sized aquarium of water holding capacity of 400 L. For experimentation fish were regularly monitored under 12 h light -12 h dark photoperiod and kept under sufficient artificial aeration system. The limnological parameters were measured regularly

Biochemical parameters of the study

The blood serum samples were processed for different biochemical parameters after naphthalene exposure and results are recorded in Table 1. The concerned fish blood protein content (PRO) was significantly reduced (F2,26 = 40.77, p <  0.001) under both low (22.67 ± 1.04 g/dl) and high (23.97 ± 1.24 g/dl) naphthalene intoxication compared with control (26.63 ± 1.32 g/dl). Lower dose recorded maximum protein reduction compared with high naphthalene exposure. The blood protein has been recognized

Conclusion

This work demonstrated the effects of naphthalene (PAH) intoxication on blood biochemistry and erythrocytes morpho-pathology of A. testudineus. Results have been able to correlate the blood biochemistry and erythrocytic morpho-pathology, and highlight the dose-specific alterations in these blood parameters. Additionally, this study addressed the necessity of linkage between these parameters and proved as the vital tools to evaluate the fish health as well as health of an aquatic ecosystem; and

CRediT authorship contribution statement

Sukhendu Dey: Conceptualization, Investigation, Formal analysis, Visualization, Writing - original draft, Writing - review & editin. Puspita Ballav: Conceptualization, Formal analysis, Resources, Writing - review & editing. Arghya Mandal: Formal analysis, Investigation, Data curation. Palas Samanta: Investigation, Formal analysis, Visualization, Writing - review & editing. Atanu Patra: Investigation, Formal analysis. Subhas Das: Investigation, Formal analysis. Arnab Kumar Mondal: Investigation,

Declaration of Competing Interest

On behalf of the authors, the corresponding authors stated that there is no conflict of interest.

Acknowledgement

Authors are indebted to Dept. of Environmental Science, The University of Burdwan providing us the infrastructure support through Laboratory facility.

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