Effect of Diazinon on the activity of amino acid in gills and stomach of Channa punctatus

International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 58 Effect of Diazinon on the activity of amino acid in gills and stomach of Channa punctatus Dr Ragini Ahirwar PG College, Ghazipur, (U.P.) India. e-mail: [email protected] ARTICLE INFO ABSTRACT ©2025 RS Publication Paper ID: IJRES69143AF4B6580 Received: 2025-10-14 Published: 2025-11-13 DOI: https://dx.doi.org /10.5281/zenodo.17 599866 Page No: 58-66 The extensive use of agricultural pesticides, such as Diazinon, has led to environmental pollution, with residues impacting aquatic ecosystems and food chains. This study investigates the effects of Diazinon on the essential amino acids arginine and tryptophan in the gills and stomach of Channa punctatus. Fish were exposed to 3.1 mg/L Diazinon for one month, with a control group maintained in tap water. Histochemical analyses, including the Sakaguchi reaction for arginine and the DMAB-nitrite method for tryptophan, were performed on gills and stomach tissues. In control fish, strong positive reactions for arginine and tryptophan were observed in gill rakers, adductor and abductor muscles, and stomach mucosa and submucosa, with moderate activity in other tissue layers. Post-Diazinon exposure, a significant reduction or absence of histochemical reactions was noted across all tissues, indicating a depletion of these amino acids. This reduction is likely due to altered membrane permeability caused by Diazinon toxicity. The findings highlight the adverse ecological and physiological impacts of Diazinon on aquatic organisms, emphasizing the need for cautious pesticide use to mitigate environmental and food chain disruptions. Key words: Stomach, Gills, Essential Amino Acid, Arginine and tryptophan . International Journal of Research in Engineering & Science Available online on http://rspublication.com/IJRES/IJRE.html ISSN:(P) 2572-4274 (O) 2572-4304 Cite This Paper: Dr Ragini Ahirwar (2025). "Effect of Diazinon on the activity of amino acid in gills and stomach of Channa punctatus". INTERNATIONAL JOURNAL OF RESEARCH IN ENGINEERING & SCIENCE (IJRES), vol. 9, no. 6, 2025, pp. 58-66. DOI: https://dx.doi.org/10.5281/zenodo.17599866 International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 59 INTRODUCTION The wide use of agricultural pesticides in discriminately has developed an ecological crisis due to environmental pollution. These pesticides residue are washed away by rain water into rivers. Most of the pesticides used have appreciately long life period and also quite stable. However, some of them undergo biodegradation on soil, water or in animals and plant bodies. Pesticides are highly toxic to the organism and poorly effected of organisms. During the second word war II introduction of DDT, a synthetic organic insecticide gave entirely a new concept in pest control. Now a day, a very large number of such insecticides belonging to different group like chlorinated hydrocarbon, organo-phosphorus compounds etc. are used. These pesticides are commonly used because of their broad-spectrum insecticidal efficiency and relative low cost. These pesticides provide a useful tool for agriculturalist and hygienist for crop protection and disease control. This chemical, if resistant in nature, prove to be still more hazardous both to plants and animals, especially in aquatic environment. The pesticide and their residue which are used in farming continuously discharge into environment, resulting to the imbalance of natural equilibrium and affect food chain. Because are part of food chain, keeping this in view the present investigation was undertaken to evaluate the impact of used pesticide on the amino acid of gills and stomach of Channa punctatus. Very little work has done on effect of pesticide on amino acid in different tissues of fish body. The present study has been designed to understand the changes in arginine and tryptophan content (essential amino acid) in the gills and stomach of Channa punctatus when treated with Diazinon. International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 60 MATERIAL AND METHODS Live specimen of Channa punctatus were collected from local agencies and were acclimatized to laboratory condition for 7 days. The fishes were divided in to two groups with 20 fishes each. The first group was kept in Diazinon solution of 3.1 mg/l. The second group served as control and was maintained under laboratory condition in ordinary tap water. The treatment was applied on fishes for one month. After one month the fishes were sacrificed one by one by decapitation. The tissues (Gills and Stomach) were removed carefully and fixed in 10% neutral formalin (as a fixative). Paraffin sections were prepared and subjected to the following histochemical tests: 1. The Sakaguchi reaction for arginine (Baker’s modification 1947) 9 . 2. DMAB nitrite method (Adam 1957) 3 . RESULTS AND DISCUSSION Arginine Gills In control fishes strong positive reaction is seen in gill raker, adductor muscle, abductor muscle and moderate activity is seen in remaining tissue. After Diazinon treatment gill raker, adductor muscle and abductor muscle shows dull activity. Details are given in table 1. Stomach In control fishes stomach showed a positive reaction in mucosa and sub mucosa, the muscularis and serosa layers showed moderate activity. International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 61 After the treatment of Diazinon dull/nil reaction is seen in all the tissue. Details are given in table 2. Tryptophan Gills Presence of tryptophan was noticed in gill raker, adductor muscle and abductor muscle in strong reaction form but moderate reaction is seen in secondary gill lamellae and primary gill lamellae. After Diazinon treatment gill raker, abductor muscle, secondary gill lamellae and primary gill lamellae shows nil activity and adductor muscle shows dull activity. Details are given in table 3. Stomach Presence of tryptophan was noticed in mucosa and sub mucosa layers of the stomach in control fishes. After diazinon treatment mucosa and sub mucosa shows dull activity. Details are given in table 4. The amino acids are essential compounds for all living cells as building blocks of protein. All the amino acid in biological system can be broadly distinguished as (1) essential amino acids are those which cannot be synthesized by living body and hence are essential to be supplied with diet, and (2) non essential amino acids are those to which the body is able to synthesize from basic ingredients and hence not essential to be present in diet. In the present study one essential amino acid, the arginine and other is tryptophan has been taken for histochemical investigation. Arginine, a constituent of most proteins, is the most common positively reacting substance in this test. A red color is produced when arginine reacts with α-naphthol and sodium hypochlorite or International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 62 sodium hypobromite in an alkaline solution. Tryptophan (α-amino-indole-3-propionic acid) was named by Neumeister for an unidentified substance found in the tryptic hydrolysate of proteins that produced a red color with chlorinated water—a reaction characteristic of indole compounds 10,16,21 . The name “tryptophan” (formerly “tryptophane”) is now generally accepted. Tryptophan is a common constituent of most proteins, although usually present in small amounts. In the present study, the p-dimethylaminobenzaldehyde (DMAB)–nitrite method has been used for the histochemical detection of tryptophan. In this reaction, tryptophan and related indolyl derivatives condense with p-dimethylaminobenzaldehyde to form a colorless intermediate, which develops a strong blue pigment upon oxidation with nitrous acid (nitrite) 3.16 . The present study provides information on the histochemical localization of the amino acids arginine and tryptophan. The diazinon pesticide has adverse effect on amino acid present in gills and stomach of Channa punctatus. Because of the limited availability of data and literature on amino acid localization, no meaningful comparison could be made. In general, a faint and depleted histochemical reaction for arginine and tryptophan was observed throughout all tissue layers following exposure to the pesticide. The observed reduction in protein content is likely due to altered membrane permeability caused by the toxicant. Present histochemical study showed that the pesticide Diazinon has adverse affect in gills and stomach 5,6,7,8,20 . Nutritional etiology of fatty liver has been recorded 12 . Lipids after carbon tetra chloride poisoning on squirrel has been recorded 18 . Diazinon has ecological risk in agricultural use 11 . It has been recorded that Diazinon treated fish showed abnormal behavior which include restlessness, arena movements, loss of equilibrium, increased opercular activities, strong spasm, and paralysis 17 . Diazinon has toxic effects on various organs on fresh water fish 1,2,4,13,15,19 . Measurement of bio-concentratation of pesticide by fresh water fish has been International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 63 recorded 14 . Thus it may be concluded that our observations may be helpful to dispel doubts concerning the reliability of a pesticide exposure to the fishes. Table 1. Distribution of arginine in the Gills of Channa punctatus after the treatment of Diazinon. +++, Very Strong Activity, ++, Strong activity, +, Moderate Activity, , Dull Activity and  Nil Activity. Table 2. Distribution of arginine in the Stomach of Channa punctatus after the treatment of Diazinon. +++, Very Strong Activity, ++, Strong activity, +, Moderate Activity, , Dull Activity and  Nil Activity. Treatment Gill raker Adductor muscle Abductor muscle Primary gill lamellae Secondary gill lamellae Control ++ ++ + + + Diazinon    _ _ Treatment Mucosa Sub mucosa Muscularis Serosa Epithelial cells Gastric glands Circular muscle Longitudinal muscle Control ++ ++ ++ ++ + + Diazinon    _ _  International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 64 Table 3. Distribution of tryptophan in the Gills of Channa punctatus after the treatment of Diazinon. +++, Very Strong Activity, ++, Strong activity, +, Moderate Activity, , Dull Activity and  Nil Activity. Table 4. Distribution of tryptophan in the Stomach of Channa punctatus after the treatment of Diazinon. +++, Very Strong Activity, ++, Strong activity, +, Moderate Activity, , Dull Activity and  Nil Activity. Treatment Gill raker Adductor muscle Abductor muscle Secondary gill lamellae Primary gill lamellae Control ++ ++ ++ + + Diazinon _  _ _ _ Treatment Mucosa Submucosa Muscularis Serosa Epithelial cells Gastric glands Circular muscle Longitudinal muscle Control + + + _ _ _ Diazinon    _ _  International Journal of Research in Engineering & Science ISSN:(P) 2572-4274 (O) 2572-4304 Available online on http://rspublication.com/IJRES/IJRE.html volume 9 Number 6, 2025 DOI: 10.5281/zenodo.17599866 Original Article ©2025 RS Publication, rspublicatio[email protected] 65 REFERENCES 1. 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