International Indexed & Referred Research Journal, April, 2012. ISSN- 0974-2832, RNI-RAJBIL 2009/29954; VoL.

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Research Paper - Zoology

Effect of Malathion and Cypermethrin on Biochemical Constituents of Freshwater Fish, Lepidocephalichthys Guntea (Ham-buch)
* Patil, M. U. ** Dr. Patole, S. S.
April , 2012 * Department of Zoology, Vimalai Uttamrao Patil College, Sakri. ** Associate professor, Department of Zoology, S. G. Patil, ASC College, Sakri. Dist- Dhule (M.S.). A B S T R A C T
In the present study, the freshwater fish, Lepidocephalichthys guntea was exposed to sub-lethal concentrations i.e. 1/4 th and 3/4th of Lc 50 values of Cypermethrin and Malathion. Both insecticides were taken for evaluation of biochemical changes in fish for 96 h of exposure. Results showed significant fluctuation in protein, decrease in glycogen and lipid over the control. These changes might be due to presence of pesticides in surrounding environment, which affect the health of ecologically important ichthyofauna in natural water bodies indicating the need to protect environment and minimize pesticide in agricultural fields.

Key words: Ichthyofauna, Biochemical constituents, Lepidocephalichthys guntea, Cypermethrin, Malathion. Introduction Acute exposures of fish to pesticides result in some biochemical changes, causing some interference. Every living organism has its own so called detoxification mechanism to get rid of foreign substances in the body, however if toxic substance are encountered in higher concentration, they are bound to bring severe adverse effects (Venkataramana et al., 2006; Satyavardhan, 2010). Such effect may be at cellular or even at molecular level but ultimately it leads to behavioral, physiological, pathological and biochemical disorders that may prove fatal (Patole et al., 2008; Rathod et al., 2009; Yaji et al., 2011). Recent understanding of different biochemical processes has proved useful in determining the mechanism of toxicity of different toxicant and also in unfolding the adaptive protective mechanism of the body to fight the toxic effect of the pollutants (Sarkar et al., 1996; Saha & Kaviraj, 2009). Besides it is also now felt that some of the biochemical alternation occurring in the body gives the first indication of the stress in the organism and hence effect on the part of the pollution (Venkataramana et al., 2006; Rathod et al., 2009). The number of changes in the biochemistry of fish was reported as result of exposure to pesticides. Presently, the aquatic ecosystem is abandoned by indiscriminate use of pesticides in agriculture field to protect the crops. Unfortunately, many non-target fresh water organisms like fish, mollusks, prawn, crabs, etc are adversely affected (Yaji et al., 2011). The biochemical studies are good parameters which help to see the effect of toxicants on metabolism of fish (Ghosh, 1986; Kajare et al., 2000). Hence, in present investigation, an attempt has been made to find out induced effect of Malathion and Cypermethrin insecticides on biochemiSHODH, SAMIKSHA

cal constituents from total body muscles of experimental fish, Lepidocephalichthys guntea. Materials And Methods The freshwater fish, Lepidocephalichthys guntea (body weight 2.0 0.5 g and length 5.0 2.0 cm) were bought from local fishermen. They were acclimatized to the laboratory conditions in well aerated and with the non-chlorinated tap water at the test medium conditions for 15 days. The physico-chemical parameter of water was analyzed prior to experiment by after APHA (1998). During this period fish were fed with standard fish diet. Injured and dead fish were removed to prevent any decomposition. A commercial grade Malathion and Cypermethrin was used for biochemical study. A stock solution of the toxicants was prepared and further diluted in ppm according to dilution technique (APHA, 1998). For experimentation, laboratory acclimatized fish were divided into five groups of 10 fish, each with average weight 15.0 2.0 g taken into glass aquaria. Group-I was kept as control and remaining groups were experimental for exposing to sub-lethal concentrations of both pesticides i.e. 1/4th and 3/ 4th dose concentration. Diet was withdrawn 24 h before experimentation. Water was renewed every 24 h in order to provide fresh oxygenated water, to maintain the concentration of pesticides and also to remove accumulated waste. All the groups were kept for 96 h. The schedule for treatment is shown below. i. Group-I: Control. ii. Group-II: Fish were exposed at 1/4th sub lethal dose (2.75ppm) to Malathion. iii. Group-III: Fish were exposed at 3/4th sub lethal dose (8.25 ppm) to Malathion.
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International Indexed & Referred Research Journal, April, 2012. ISSN- 0974-2832, RNI-RAJBIL 2009/29954; VoL. IV * ISSUE-39

Table 1. Biochemical changes in fish, Lepidocephalichthys guntea exposed to sub-lethal doses of Malathion and Cypermethrin for 96 h Parameter Control Malathion Cypermethrin (Group-I) LC 50 ) LC 50 LC 50 LC 50 (Group-II (Group-III) (Group-IV) (Group-V) Glycogen 108. 25 0. 11 79. 413 0. 16 67. 16 0. 21 61. 81 0. 16 54. 44 0. 16 (- 26. 86) * (- 37. 8)** (- 42. 60) ** (- 50. 00) ** Protein 160. 28 0. 27 149. 30 0.20 115. 68 0. 27 106. 26 0. 11 98. 87 0. 12 (- 6. 80) NS (- 28.04)* (- 33. 75) ** (- 38. 62) ** Lipid 11. 26 0. 80 8. 19 0. 10 7. 55 0. 22 8.2 0. 14 3. 72 0. 17 (- 27. 20) * (- 31. 80) * (- 27. 18) * (- 66. 37) *** All values expressed in mg/ 100 g wet weight tissue and mean S. D. of six observations. * Significant at P< 0.05, ** P < 0.01, *** P < 0.001. NS = Non significant at P > 0.05. * Values in parentheses indicate percentage change over control (taken as 100 %).

iv. Group-IV-Fish were exposed at 1/4th sub lethal dose (1.3 ppm) to Cypermethrin. v. Group-V-Fish were exposed at 3/4th sub lethal dose (3.9 ppm) to Cypermethrin. At the end of 96 h treatment, those which survive were removed from test media, blotted on paper and sacrificed. For further study, the freshly isolated body muscles were used. They were homogenized and then centrifuged at 3000 rpm for 10 minutes. Supernatant were used for estimation of glycogen and protein, whereas for lipid estimation, the body tissues were subjected to ethyl ether: ethanol solvent (3:1). Mean, standard deviation and the student't' test were adopted to evaluate the level of significance. The standard procedures were used for the estimation of glycogen (Miller, 1972), protein (Lowry et al., 1951) and lipid (Bling & Dyer, 1959). Results and Discussion The biochemical changes are presented in Table 1. The impact of metabolic process may result in either increase or decrease in the metabolic rate, also depending on the site of action. The chemicals, that monitoring the metabolic rate during exposure to the toxicant can provide valuable information regarding physiological and biochemical mechanisms activated by the chemicals. Depletion of glycogen content was noted in exposed fish. As compared to Malathion, significant depletion in glycogen content was found with Cypermethrin. A fall in glycogen level, in exposed fish indicates its rapid utilizes to meet the enhanced energy demand through; glycolysis pathway (Ghosh, 1986; Satyavardhan, 2010) or it might be due to over activity of muscle under Pesticidal stress (Baigh et al., 1991). It is known that pesticides act on endocrine system. Hence, decreased in glycogen content is due to inhibitor of hormones which contribute to the glycogen synthesis decrease. Glycogen serves as primary source of energy for metabolic process in various organisms under stressful environmental condition (Singh & Gupta, 2007). The disturbance in the glycogen profile

is one of the most outstanding biochemical lesions due to action of many chemicals (Kajare et al., 2000; Saha & Kaviraj, 2009). Significant decrease in protein was observed in groups exposed to 3/4th Malathion (i.e.28.04 mg), 1/ 4th Cypermethrin (i.e. 33.75 mg) and 3/4th Cypermethrin (i.e. 38. 62 mg), whereas it was non-significant (i.e.6.8 mg) in group exposed to 1/4th LC50 concentration of Malathion. Lepidocephalichthys guntea on acute expose to sub- lethal concentration showed low protein level of whole body tissues as compared to control and also fluctuated with the exposure at 96 h. The sublethal concentrations of Malathion were chosen to evaluate their toxic impact on the protein metabolism. Reduced level of protein could be due to the reduction in protein synthesis because of liver cirrhosis (Parate & Kulkarni, 2003), which signifies the effect of toxicants or pollutants on organism. Protein metabolism is also considered being one of the physiological events involved in the compensatory mechanism under stress condition (Devi & Piska, 2006; Venkataramana et al., 2006; Yaji et al., 2011). The level of lipid content decreased significantly in all treated groups, when compared with control. Maximum decrease of lipid was observed in group exposed to 3/4th Cypermethrin (i.e.66. 37 %). In other groups these are 27.20 %, 31.8 % and 20.18 % at 1/4th Malathion, 3/4th Malathion and 1/4th Cypermethrin respectively. Lipid plays as important role in energy metabolism and provide energy to metabolic processes. They are also important for cellular and sub-cellular membrane. It is used as energy reservoir, stored and transported in the form of glycerol esters (Patole et al., 2008; Rathod et al., 2009). These decreases in total lipid content of whole body tissue at different concentrations suggest that the lipid might have been channelized to meet the metabolic demand for the extra energy needed to mitigate the toxic stress. Hence, lipid level decreases significantly because liver is the principle site of detoxification in vertebrates (Dixit, 2005). In nutshell, both pesticides Malathion and

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Cypermethrin are highly toxic to Lepidocephalichthys guntea at acute exposure leading to affect the nutritive value of the fish as well as the entire metabolism studied is found to be sensitive change which reflects changes in normal activities of various functional systems. It is concluded that, such toxicants induce alteration in biochemical composition of fish muscles under Pesticidal stress. It might be due to glycogenolysis, pro-

teolysis and lipolysis to meet the energy demand. Acknowledgements Authors are thankful to principal, S. G. Patil College, Sakri for providing necessary laboratory facilities. Thanks are due to the Director, Zoological survey of India, (WRO, Poona) for valuable think in identification of experimental fish.

R E F E R E N C E
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