TOXICITY OF PROPARGITE ON CHEMICAL COMPOSITION AND FATTY ACID PROFILE IN

In this study, effects of propargite is an organosulfiteacaracide/miticide pesticide were investigated in Channa striatus , muscle and liver chemical composition, the total fatty acid profile and free fatty acid. Fish were exposed to sub lethal concentration as control, 1ppm, 2ppm of 15 days and 30 days of propargite. As a result of a study, chemical composition of muscle and liver of Channa striatus exposed moisture, crude protein and ash were significantly (P>0.05) different in the propargite concentration of 1ppm and 2ppm of 30 days then compared to 15 days and control. Estimation of muscle and Liver tissue saturated fatty acids (SFAs) as palmitic acids were increased in propargite 2ppm of 15 days while compared to control. Correspondingly,

1053 muscle and liver transferred in to mark sterilized polythene bags and stored in a freezer at 20°C until further analysis.

Fatty acid methyl esters preparation and Gas chromatography
During fatty acid analysis, each liver and muscle samples were freeze dried (lyophilized) and oven dried at 67°C for 24h. Then it was grounded finely with pestle and mortar. The analysis of fatty acid methyl esters (FAMEs) from these muscle and liver samples were performed by standard procedures. To 50 mg of muscle and liver samples were added to 1gm of 1.2M NaOH in 50% methanol with glass beads (3mm dia) in a screw-cap tube and then incubated at 100°C for 30 min in a water bath. The saponified samples were cooled at room temperature for 25 min, they were acidified and methylated by adding 2 ml 54% 6N HCl in 46% methanol and incubated at 80°C for 10 min in water bath. After rapid cooling, methylated FAS were extracted with 1.25 ml 50% methyl-tert butyl ether (MTBE) in hexane. Each sample was mixed for 10 min and the bottom phase removed with a pasteur pipette. Top phase was washed with 3ml 0.3M NaOH. After mixing for 5 min, the top phase was removed for analysis. Following the base wash step, the FAMEs were cleaned in anhydrous sodium sulphate and then transferred in to GC sample vial for analysis. FAMEs were separated by gas chromatograph (HP 6890 N, Agilent Technologies, USA). FAMEs profiles of the samples were identified by comparing the commercial Eucary data base with MIS Software package (MIS Ver. No. 3.8, Microbial ID. Inc., Newark, Delaware) (Bligh and Dyer, 1959).

Statistical Analysis
All the dates were subjected to one way ANOVA using statistical software of SPSS version 16.0. Duncan"s Multiple Range test was used to determine the difference among treatment means at 5% level of significance.

Muscle and liver chemical composition
The effect of sub-lethal exposure of propargite on moisture, fat, crude protein and ash levels in liver and muscle of C.striatus, at different periods of exposures were presented in table 1 and 2. The muscles composition of moisture, crude protein and ash were significantly decreased in the propargite concentration of 1ppm and 2ppm of 30 days (62.81±0.74, 50.36±0.80) then compared to 15 days (65.34±0.75, 53.24±0.82) and overall muscle composition were decreased in all treated group when compared to control (82.30±0.66), respectively (Table1).
The chemical composition of liver such as moisture, crude protein and ash were significantly decreased in the propargite concentration of 1ppm and 2ppm of 30 days (60.15±0.73, 28.18±0.59) then compared to 15 days (64.32±0.75, 39.20±0.74) and overall muscle composition were significantly decreased in all treated groups when compared to control (78.60±0.76), respectively ( Table 2).
Composition of fat level such as muscle and liver of C.striatus, at different periods of exposures were significantly increased in all treated groups when compared to control, respectively (Table 1 and 2).

Profile of Fatty acids analysis through gas chromatography (GC)
Profile of fatty acid and free fatty acid levels in liver and muscle of C.striatus, at different periods of exposures were presented in table 3, 4, 5 & 6 and figure 1 & 2. Values of muscle tissue saturated fatty acids (SFAs) of Capric acid, Undecanoic acid, Lauric acid, Myristic acid and stearic acid (1ppm 15days) were higher in propargite 1ppm of 30 days, as well as palmitic acid were increased in propargite 2ppm of 15 days when compared to other concentration treated groups and control, ( Table 3).

Free fatty acid profile in muscle tissue
Mono Unsaturated FAs of Elaidic acid and cis-11-Eicosenoic acid were not detected in all treated groups then control, as well as cis-10-pentadecenoic acid and Oleic acid were increased in propargite 1ppm of 15 days and 30 when compared to other groups and control (Table 3).
Poly Unsaturated FAs as Linolelaidic acid, Linoleic acid, Cis-5, 8, 11, 14, 17-Eicosapentaen acid and Cis-4, 7, 10, 13, 16, 19-Docosahexaenoic acid were gradually increased in all treated groups when compared to control. Concerning to PUFAs, alpha-Linolenic acid was not detected in all treated groups then control (Table 3 and fig 1). Total amount of muscle tissue of MUFAs and PUFAs were decreased in all treatment groups compared to control (Table 4 and fig 1).

Free fatty acid profile in liver tissue
Saturated fatty acids (SFAs) of Liver tissue such as Caprylic acid and Arachlic acid were dominant in propargite 2ppm of 15 days whereas the palmitic acid (C16:0) and stearic acid (C18: 0) levels were decreased when compared to control.

Discussion:-
The present study constitutes the pesticide effect of sub-lethal exposure of propargite on chemical composition and fatty acid activities of C. striatus. The present study propargite chemical composition of muscle and liver tissues in the results moisture contents, while treatment groups had significantly decrease. However, the energy values were changes in the muscle tissue moisture content in chlorpyrifos treatments, in agreement this content in monocrotophos-exposed juvenile Indian carp Labeo rohita (Ramaniet al., 2002).
In the present study the fat content was found to be muscle and liver tissues in the results as treatment groups had significantly increased. Gluer et al., (2008) reported that fat level attributed the rise might be due to disruption in the hepatic cell owing to stress induced toxicants and thereby releasing cholesterol to blood. Higher fat level in muscle might due to accumulation of cholesterol in the tissue (Firatet al., 2011) Jankowskaet al., (2010) and Valfreet al., (2003).
The results of the present study showed that crude protein of muscle tissues has significantly decreases (Onyelikeet al., 2000). Protein content also due to the rapid utilization of tissues as protein decreases when the animals were under stress conditions. It has been shown that ash muscle tissues decrease (Abiiet al., 2007). Liver ash contents increase indicated that Alterations in moisture and ash contents have been suggested to be due to the reduction of food consumption and food conversion efficiency under stress (Nair &Sherief 1998).
The results of the present study showed that free fatty acid profile in muscle tissue were found that propargite exposure the hepatic fatty acids profile, the proportion of monounsaturated fatty acids (MUFA) increased, saturated fatty acid (SFA), and polyunsaturated (PUFAs) acids decrease was respectively. The n-3/n-6 ratio, the content of Eicosapentaenoic acid (EPA) and Docosahexaenoic acid (DHA) were preventive effects on human coronary artery disease (Leaf & Webber, 1988). The presence of docosahexaenoic acids (DHA) in all fish species from the Indus River suggests that these fish species can have a healing effect to alleviate muscle pain and inflammation. Therefore, fish have been suggested as a key component for a healthy diet in humans (Rahmanet al., 1995). Significant levels of EPA and DHA in fish species of this study indicated that these species can be used to supplement essential fatty acids in the human diet. Although the EPA and DHA percentages in the examined fish species muscle total fatty acids were low, they were found in significant levels in these fish species muscles, due to the large percentage of fat in the analyzed fish species. Changes according the influence of nutrients body composition have also on other major carp"s rohu (Umeret al., 2011).
The patterns of fatty acid profile of fish compared well with those observed by (Hashimet al., 2007). However, the fatty acids compositions of the muscle cell membranes were especially important factors in determining the stability because oxidative changes were initiated from the membrane components of muscle (Buckley et al., 1989).
The results of the present study showed that Fatty acids profile in muscle tissue of C. striatusexposed to sub-lethal concentration of propargite among saturated (SFA) fatty acids of fish as palmitic acid (16: 0) increased those reported by previous study was comparable with fish in curing illness to improve the health process (Luczynskaet al., 2008). It was reported that palmitic acid was predominant in fresh water channel catfish Icataluruspunctatus (Sathivel et al., 2002)). According to the palmitic acid increases the risk of developing cardiovascular diseases (WHO, 2003), indicating that it may increases LDL levels in the blood.

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However, in the present study, Oleic acid were increased Ackman, (1980) and Kolakowskaet al., (2002) reported that monounsaturated fatty acids (MUFA) which good agreement with the present oleic acid were dominate in all fresh water fish of C.carpio, L.rohita and O. mossambicus respectively. Oleic acids along with other monounsaturated fatty acids in red blood cell membranes were positively associated with breast cancer risk (Andrea Micheliet al., 2001).
Among the polyunsaturated fatty acids (PUFAs) as docosahexaenoic acid (DHA, C22: 6n3) and eicosapentaenoic acid (EPA, C20: 5n3) were dominant (Ackman, 1986). DHA were as major component of the brain, retina, muscle and heart, plays a vital role in brain and eye development. The eicosanoids derived from EPA were positive effects, such as vasodilation and anti aggregation (Reilly et al 1998). DHA represents an extreme of the omega -3 fatty acids and linked in a positive way to an enormous variety of human afflictions including cancer and heart disease, as well as to neurological and brain development (Stillwell, and Wassall, 2003). The most important factor affecting the pesticide of the quality of fish muscle is the percentage n-3 FA such as EPA and DHA.
The present study showed that propargite exposure liver influences the hepatic fatty acids, especially the saturated fatty acids (SFAs), monounsaturated (MUFAs) and polyunsaturated acids (PUFAs) were decreased. Montero et al., (1999) reported that stearic acid and oleic decreased remained almost same at the endosulfan exposure reduction in unsaturated fatty acid (USFAs) in liver could due to their utilization for energy purpose. Many nutritional of lipids based on the proportions reported that Jankowskaet al., (2010) and Valfreet al., (2003).

Conclusion:-
The overall results demonstrated that sub-lethal exposure to propargite had significant impact of affect on lipid and total free fatty acid profile of C. striatus. The analysis of seasonal as well as annual variations of the FAs profiles of the fishes deemed by this study as most suitable sources of PUFAs and MUFAs, then the knowledge of the relative abundance of each fish species in different areas. This study provided new clue, it"s that further investigation about the physiological effects of major enzyme activities. This study reiterates the important of judicial use of pesticide, in order to avoid to contamination of fresh water bodies.