Profenofos induced Protein Alterations in Fresh water crab, Paratelphusa jacquemontii (Rathbun)

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1 Profenofos induced Protein Alterations in Fresh water crab, Paratelphusa jacquemontii (Rathbun) A.Maharajan, R.Saraswathi, P.Kumarasamy PG & Research Department of Zoology, Khadir Mohideen College, Adirampattinam , Thanjavur Dist, Tamil Nadu, India. Abstract Profenofos, a well-known organophosphate pesticide has been in agricultural use over the last two decades for controlling pests of paddy, cotton and tobacco. In the present study, an attempt has been made to study the effect of profenofos on protein content of different tissues like muscle (MU), hepatopancreas (HP) and gill (GL) of the freshwater crab, Paratelphusa jacquemontii (Rathbun). The crabs were exposed to sub lethal concentration (1/10 th & 1/20 th of 96 h LC 50 ) of profenofos for a period of 15 days. The treatments of these profenofos brought about significant decrease in protein content in all tissues as compared to control. The maximum decrease was observed in muscle. The percent decrease of total protein content was in the order of: muscle > hepatopancreas > gills. The obtained results indicate that profenofos has more toxic. The significance of these studies as bioindicator for assessing the toxicity and economic importance of the crab are discussed. Key words: Profenofos, Toxicoty, Protein, Paratelphusa jacquemontii Introduction With rapid industrialization and increase in human population, the pollution of water bodies has become a universal phenomenon in the present day world 1. The important sources of water pollution are industrial effluent, domestic, sewage, drainage and pesticides, which pollute the river and major water sources 2. Profenofos, a well-known organophosphate pesticide has been in agricultural use over the last two decades for controlling pests of paddy, cotton and tobacco. Profenofos has been classified as a moderately hazardous (Toxicity class II) pesticide by the World Health Organization (WHO) and it has a moderate order of acute toxicity following oral and dermal administration. Profenofos is extremely toxic to fish and macro invertebrates. The acute toxic action of profenofos is the inhibition of the acetylcholine esterase activity resulting in toxicity also in humans. Microbial degradation of organophosphate pesticides is of particular interest because of the high mammalian toxicity of such compounds and their widespread and extensive use. The most significant step in detoxifying organophosphate compounds is hydrolysis since that makes the compounds more vulnerable to further degradation. The enzyme responsible for catalyzing this reaction is referred as an esterase or phosphotriesterase. The nutritional value of different species of fish and shellfish depend on their biochemical components such as protein, carbohydrate and lipids. These proximate components could serve as sensitive indicators for detecting potential adverse effects, particularly the early events of pollutant damage because their alterations appear before the clinical symptoms produced by the toxicant 3. It is therefore important that potential effects of acute and chronic concentrations of pollutant on proximate composition are determined and interpreted to delineate mechanisms of pollutant action and possibly ways to mitigate adverse effects 4. Presence of pesticide in streams and lakes is largely due to the runoff from agricultural fields and outfall from pesticide manufacturing factories 5. Pesticides are not highly selective but are generally toxic to many macrophytes, non-target organisms such as fish 6. Fish is highly nutritious, easily digestible and much sought after food. Nutritional value of fish depends on their biochemical composition, which is affected by the water pollution 7. Alterations in biochemical components as response to environmental stress are authenticated by many authors. Tilak et al. 8 studied the toxicity and effect of chloropyriphos to the freshwater fish, Labeo rohita (Hamilton); Arockia and Mitton 9 investigated the effect of carbamate pesticide lannate (methomy1) on the biochemical components of the freshwater cichlid, Oreochromis mossambicus (Peters); Impact of Cypermethrin and Ekalux on respiratory and some biochemical activities of a fresh water fish, Tilapia mossambica was studied by Logaswamy and Remia 10. The present investigation of toxicity of profenofos in Paratelphusa jacquemontii is carried out. Since, it is cultured in the ponds of Delta Districts of Tamil Nadu. The culturing pond receives river runoff from paddy fields has the possibility to contain pesticides. ISSN : X Vol 3 Issue 3 Jun-Jul

2 Materials and Methods Test animal collection and maintenance Fresh water crab, Paratelphusa jacquemontii of carapace size ranging from 4-5cm and weight 50-70g were collected from the paddy field of Orathanadu, Thanjavur Dist, Tamil Nadu. They were transported and kept for acclimatization in rectangular tank of 100 l capacity containing well aerated filtered fresh water maintained at ambient temperature (27±2 C) for a period of one week. Before stocking, the tank was washed with clean water several times. Finally, the tank was washed with 0.1% KMnO 4 for disinfection. Before introducing into the tank, the fishes were screened for any visible pathological symptoms and were treated with 0.1% of KmNo 4. Preparation of stock solution for Profenofos toxicity test 1 ml of Profenofos was dissolved in one litre of double distilled water and used as the stock solution for preparing different concentrations of Profenofos in rearing water. It was stored in a clean standard flask at room temperature, in the laboratory. Exploratory test Exploratory tests, otherwise called range finding test, were carried out to assess the approximate effective concentration range of profenofos required for conducting short term tests to assess the effect of profenofos on the metabolic function of the crab, as recommended by APHA 11. The test solutions were prepared over a wide range of concentrations. These tests were performed by exposing 10 specimens of Fresh water crab, Paratelphusa jacquemontii in 10 litre fresh water containing different concentrations of profenofos. The dead animals were removed immediately. Death of each animals was recorded. Three replicates were made for short term toxicity tests, the least concentration was chosen where no mortality was recorded in 24hrs and the highest lethal concentration was where 100% mortality was recorded in 24hrs. Acute Toxicity test To study the toxicity of profenofos, the Static Bioassay Method 11 was followed. The test individuals were exposed to selected and serially diluted profenofos concentrations. For acute toxicity test, 10 active animals each were exposed to various concentrations of the profenofos (0.005,0.010,0.020,0.030,0.040,0.050,0.060 and ppm) using fresh water as control. The manifestation time and survival time of crab were observed. Crabs were exposed to the above said concentrations along with common control. Experimental animals were starved for one week. The experiments were conducted in three replicates at room temperature. No feed was given during the test period. Sub lethal toxicity tests For sublethal toxicity tests, the crabs were grouped into three batches. Each batch had 10 animals and had 3 replicates. Group: I Crabs were maintained in normal Fresh water and served as control. Group: II Crabs were exposed to the sublethal concentration of ppm (1/10 th of LC 50 value for 96 hours) of profenofos in Fresh water. Group: III Crabs were exposed to the sublethal concentration of ppm (1/20 th of LC 50 value for 96 hours) of profenofos in Fresh water. The media were renewed every alternate day. Crabs were fed daily with artificial feed. Two specimens each from the groups I, II and III were sacrificed after 0, 5 th 10 th and 15 th days of the experiment. After their respective exposure period, tissues like muscle, Hepatopancreas and gills were collected on ice and used for the analysis of protein by the method of Lowry et al. 12. Bovine serum albumin was used as standard. The amount of total protein content was expressed in terms of mg % of wet weight of tissue. Each observation was confirmed by taking at least three replicates. The significance of difference in protein values control and experimental animals was tested by student t test 13. ISSN : X Vol 3 Issue 3 Jun-Jul

3 Results and Discussion Acute toxicity test Acute toxicity study was done to find out the impact of profenofos on Paratelphusa jacquemontii within a short period. In the present study the 96hrs LC 50 value was found to be 0.038ppm. Among the test concentrations prepared from the preliminary toxicity test the mortality of 50% of the population after 96hrs exposure was observed on 0.038ppm concentration of profenofos. Changes in total protein of different tissues Levels of the TP in different tissues of control and exposed crabs during the exposure period are depicted in Fig.1,2 &3. The TP concentrations were significantly lower in test crabs than those of controls on all DoE (P<0.01). The rate of depletion was found to be highly time and tissue dependent. The order of percent decrease of the TP concentrations in different tissues at the end of 15 DoE was observed to be MU>HP>GL. A progressive depletion in the TP levels of test crabs was recorded in the tissues of GL and MU during the exposure period. Significant variation in the TP content between exposure concentrations of and ppm was noticed (P>0.01). The levels of hepatic protein of test crabs were found to be almost similar to that of control crabs on 0 and 5 DoE but depletion was more prominent on 10 and 15 DoE (Fig.1). The magnitude of depletion in the hepatic protein was directly proportional to the concentration of profenofos. Higher percent depletion in the hepatic protein was observed in test crabs exposed to ppm compared to those exposed to ppm of profenofos(p<0.05). Indiscriminate and wide spread use of toxic substances like heavy metals, organ tin compounds, metallic pollutants etc. in variation controlling products are very common practice. These agents adversely effects on many non-target organisms like fishes, bivalves, prawns, crabs, etc. of the aquatic ecosystem. These environmental pollutants bring about damage to different organs and disrupt the physiological and biochemical processes in the organisms. Many of the toxic substances remained persistent and non-biodegradable with their residues lasting in our environment. Depletion in protein level in different tissues of Paratelphusa jacquemontii were observed consistent decrease as exposure period and lethal concentration increases. The maximum decrease recorded at ppm in hepatopancreas followed by gill and muscle for all concentrations. All the tested tissue showed significant decreased of protein level when compared to control tissues of Paratelphusa jacquemontii. The results obtained in the present finding were in agreement with some researchers who have showed depletion in protein content in the different tissue exposed to acute and chronic concentrations of different pollutants. The decrease in protein content was reported in freshwater prawn Macrobrachium kistnensis exposed to some pesticides 14. Shivprasad Rao. et al. 15 studied the effect of methyl parathion pesticide on Pila ghobusa and reported that the depletion in protein level in different tissues due to enhanced proteolytic enzyme activity and decreased protein synthesis. Similar results were reported by some researchers, 16,17. The decrease in protein along with an increase in the levels of amino acids and increase in the levels of RNA might indicate an increase catabolism of protein and decrease synthesis reported by Indra and Ramalingam 18. Conclusion In present investigation, reduction in total protein content was noted in the muscle, gill and hepatopancreas, of the exposed to Paratelphusa jacquemontii different concentrations of profenofos. This was possibly due to stress condition caused by toxicity of profenofos on protein metabolism or due to enhanced proteolytic activity as a consequence of increased metabolic demands following exposure to the toxic stress of profenofos. From the above discussion and all the available literature, we can conclude that the profenofos is very toxic to the freshwater crab, Paratelphusa jacquemontii. Therefore the release of organotin compounds in aquatic environment especially in freshwater ecosystem might be controlled. This type of study is useful to compare the sensitivity of various species of aquatic animals and potency of profenofos using LC 50 values and to derive safe concentration. References [1] Bela, Z., Prasad R., Impact of pollution on fresh and marine water resources. J. Poll. Res 2008, 273, [2] Maruthanayagam, C., Sharmila, G., Haematobiochemical variations induced by the pesticide, Monocrotophos in Cyprinus carpio during the exposure and recovery periods. Nat. Environ. Poll. Tech 2004, 3, [3] Rao, J.V., Toxic effects of novel organophosphorus insecticide (RPR-V) on certain biochemical parameters of euryhaline fish, Oreochromis mossambicus. Pestic. Biochem. Physiol 2006, 86, [4] Matos, P., Fontaınhas-Fernandes, A., Peixoto, F., Carrola, J., Rocha, E., Biochemical and histological hepatic changes in Nile tilapia,oreochromis niloticus exposed to carbaryl. Pestic. Biochem. Physiol 2007, 89, [5] Anandkumar, S., Effect of endosulphan on oxygen consumption of Lepidocepalicthys thermalis (bleeker). Comp. Phys. Ecol 1988, 23, [6] Ayoola, S.O., Toxicity of glyphosate herbicide on Nile tilapia (Oreochromis niloticus) juvenile. Afr. J. Agric. Res 2008, 3 (12), ISSN : X Vol 3 Issue 3 Jun-Jul

4 [7] Prado, R., Rioboo, C., Herrero, C., Cid, A., The herbicide paraquat induces alterations in the elemental and biochemical composition of nontarget microalgal species. Chemosphere 2009.,76, [8] Tilak, K.S., Veeraiah, K., Ramanakumari, G.V., Toxicity and effect of chloropyriphos to the freshwater fish, Labeo rohita (Hamilton) and Tilapia Mossambica. Recent Res. Sci. Technol 2001, 1, 4-7. [9] Arockia, J.J., Mitton, J.M.C., Effect of carbamate pesticide lannate (methomy1) on the biochemical components of the freshwater cichild, Oreochromis mossambicus (Peters). Ind. J. Environ Ecoplan 2006, 12, [10] Logaswamy, S., Remia, K.M., Impact of Cypermethrin and Ekalux on respiratory and some biochemical activities of a fresh water fish, Tilapia mossambica. Current Biotica 2009, 3, [11] APHA., Standard methods for the examination of water and waste water APHA, AWWA and WPCF, New York [12] Lowry, O.H., Rosenbrough, N.J., Randall, R. J., Protein measurements with folin phenol reagent. J. Biol. Chem 1957, 193, [13] Mungikar, A. M., Biostatistical Analysis. Saraswati Publ. Printing Press, Aurangabad. pp. 2003, [14] Nagbhushnam, R., Deshpande, J., Sarojini, R., Effect of some pesticides on the biochemical constituents of freshwater prawn, Macrobrachium kistnensis. Proc. Nat. Symp. Ecotoxicol 1972, [15] Shivprasad Rao., Sathyaprasad, K., Madhu, C.H., Ramana Rao, K.V., Effect of methyl parathion on tissue protein and excretory product of the snail, Pila globosa (Swainson). Nat. Acad. Sci. Letters 1981, 4, [16] Vincent, S., Ambhore, T., Kumar, L.C.A., Selvanayagam, M., Biochemical response of the Indian major carp, Catla catla (Ham) to chromium toxicity. Indian J. Environ. Health 1995, 37(3), [17] Geraldine, P., Saravanabhavan, P., Kalia Murthy, T., Zayappragassarzan, Z., Effects of dichlorvos intoxication in the freshwater prawn, Macrobrachium malcolmsonii. J. Environ. Biol 1999, 20, [18] Indra, D. and K. Ramalingam., Injury stress on compounds on the physiologylogy of freshwater tissue sugar protein metabolites concentration phosphate activity in Achatina fulcia. Nat. Acad. Sci. Letters, 19(1): ISSN : X Vol 3 Issue 3 Jun-Jul

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