ISSN: 2084-3577

TMKARPISKI
PUBLISHER

Journal of Biology and Earth Sciences ORIGINAL ARTICLE

BIOLOGY

Influence of synthetic and plant extracts on biochemical changes in the gonadal tissue of snail Lymnaea acuminata
Vatsala Singh, J.N. Tiwari
Department of Zoology, TD College Jaunpur, UP, India

ABSTRACT

Fasciolosis is now recognized as an emerging human disease. Control of intermediate host is an effective tool for control of liver fluke infection. In present study effect of sublethal treatment (20% and 60% of 24h LC50) with permethrin, cypermethrin, Allium sativum and Abrus precatorius plant extract caused a significant (p<0.05) reduction in the protein, amino acid, DNA and RNA level in the gonadal tissue of snail Lymnaea acuminata. Maximum reduction in the level of protein (33.55% of control), amino acid (1 6.21 % of control), DNA and RNA (32.96 % and 1 3.70% of control) with the treatment of 60% of 24h LC50 of permethrin after 96h exposure periods. Among plant extract Abrus precatorius is more pronounced than Allium sativum.

Key words: Fasciolosis; Biochemical parameters; Snail; Permethrin; Cypermethrin; Plant extract. J Biol Earth Sci 201 3; 3(2): B1 64-B1 68

Corresponding author:

J.N. Tiwari Department of Zoology, TD College Jaunpur, UP, India. E-mail: vatsalasingh74@gmail.com
Original Submission: 11 May 201 3; Revised Submission: 1 0 June 201 3; Accepted: 1 7 June 201 3 Copyright 201 3 Author(s). Journal of Biology and Earth Sciences 201 3 Tomasz M. Karpiski. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

http://www.journals.tmkarpinski.com/index.php/jbes or http://jbes.strefa.pl e-mail: jbes@interia.eu

Journal of Biology and Earth Sciences, 201 3, Vol 3, Issue 2, B1 64-B1 68

B1 64

Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata

INTRODUCTION
Fasciolosis is an important helminth disease caused by two trematodes Fasciola hepatica and Fasciola gigantica [1 , 2]. The definitive host range of Fasciola is very broad and includes many herbivorous mammals, including humans. The fresh water snails Lymnaea acuminata and Indoplanorbis exustus are intermediate host of this parasite [3, 4]. Fasciolosis is now recognized as an emerging human disease: The World Health Organization has estimated that 2.41 7 million people are infected with liverfluke, and a further 1 80 million are at risk of infection [5]. Annual losses are estimated to be at least three million dollars [1 , 6]. Extensive use of synthetic molluscicides are cause serious environmental hazards. Alternatively, A number of chemically diverse plant molluscicide has been used as molluscicides as and more effective than synthetic [7]. Advantages of natural product over synthetic have eco-friendly, biodegradable abidance are less likely to accumulate in the environment. Recently, Singh and Tiwari, [8] reported that the synthetic molluscicides and extracts of plants caused a significant reduction in the fecundity, hatchability and survival of the young snails. In the present study describe the molluscicide of synthetic and extract of plants against certain biochemical changes in the gonadal tissue of snail Lymnaea acuminata.

24h the biochemical estimations were made in the gonadal tissue of the experimental snail. Treated/control snails were removed from the glass aquaria and their gonadal tissue were dissected out, the adherent tissue was removed and the organ was put on a filter paper for absorption of water. 1 5-20 snails had to be dissected in order to obtain 1 00 mg nervous tissues.

Plants used

Plants used in this works were collected locally. These were identified at the herbarium of the Botany Department, DDU Gorakhpur University, Gorakhpur where voucher herbarium specimens (# No 3260 for Allium sativum and # 3829 for Abrus precatorius) are on deposit.

Estimation of total protein and free amino acid

Total protein (g/mg) was estimated according to Lowry et al., [9] using bovine serum albumin as a standard. Homogenates of gonadal tissue were prepared in 1 0% (w/v) trichloroacetic acid (TCA). Total free amino acids (g/mg) were determined according to the method of Spies [1 0].

Estimation of nucleic acids

MATERIALS AND METHODS Test animals

Adult snails L. acuminata (2.250.20 cm in length) were locally collected from ponds, pools, lakes and low-lying submerged area in Jaunpur, U.P., India. The snails were acclimatized for 72 hours in dechlorinated tap water. The pH of the water was 7.1 -7.3 and dissolved oxygen, free carbon dioxide and bicarbonate alkalinity were 6.57.2, 5.2-6.3, and 1 02.0-1 05.0 mg/l, respectively. Snails were exposed to sublethal concentrations (20% and 60% of 24h LC50) of different synthetic (Permethrin, Cypermethrin) and Allium sativum bulb and Abrus precatorius fruit plants molluscicides. Six batches were prepared for each concentration. Control aquarium contained only dechlorinated tap water without treatment. Different biochemical assay were performed viz. protein, amino acid, DNA and RNA in gonadal tissue of L. acuminata. After

DNA and RNA in gonadal tissue of L. acuminata were estimated according to Schneider [11 ] using diphenylamine and orcinol reagents, respectively. Homogenates (1 .0 mg/ml, w/v) were prepared in 1 0% TCA at 90C and centrifuged at 5000 g for 20 minutes. The supernatants were used for DNA and RNA estimations.

Statistical analysis

Each experiment was replicated at least six times. Values of biochemical parameters protein, amino acids, DNA and RNA were exposing as mean SE. Student t-test was applied locate significance changes in between control and treated groups Sokal and Rholf [1 2].

RESULTS
There was a significant (p<0.05) decrease in protein, amino acid, DNA and RNA level in the gonadal tissue of snail L. acuminata exposed to 20% and 60% of 24h LC50 of permethrin, cypermethrin, A. sativum and A. precatorius were snail removed from 24h and 96h exposure period. Permethrin and cypermethrin caused maximum

Journal of Biology and Earth Sciences, 201 3, Vol 3, Issue 2, B1 64-B1 68

B1 65

Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata
Effect of 24h exposure to sublethal concentrations (20% and 60% of 24h LC50) of synthetic and plants molluscicides on the level of protein (g/mg), amino acid (g/mg), DNA and RNA (g/mg) in the gonadal tissue of Lymnaea acuminata.
Table 1.

Each value is mean SE of six replicates. Values of parentheses are per cent change with control taken as 1 00%. *Significant(p<0.05) when t-testwas applied in between treated and control groups. Effect of 96h exposure to sublethal concentrations (20% and 60% of 24h LC50) of synthetic and plants molluscicides on the level of protein (g/mg), amino acid (g/mg), DNA and RNA (g/mg) in the gonadal tissue of Lymnaea acuminata.
Table 2.

Each value is mean SE of six replicates. Values of parentheses are per cent change with control taken as 1 00%. *Significant (p<0.05) when t-test was applied in between treated and control groups. Journal of Biology and Earth Sciences, 201 3, Vol 3, Issue 2, B1 64-B1 68

B1 66

Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata

reduction of protein, amino acid, DNA and RNA level in the gonadal tissue of snail L. acuminata than the A. sativum and A. precatorius. 24h treatments with 20% and 60% of 24h LC50 of permethrin caused maximum reduction in protein level (40.75 % and 36.59 % of control), whereas, 96h treatment caused maximum reduction in (38.04 %, 33.55 % of control) the level of protein. Snail exposed to 20% and 60% of permethrin caused maximum reduction in total free amino acid (24h treatment -8.68 % and 7.34 % of control) and (96h treatment -6.49 % and 5.96 % of control) (Table 1 and 2). Maximum reduction 24h in the level of DNA (27.92% of control) treatment with 60% of 24h LC50 of permethrin. Whereas, RNA level in gonadal tissue of snail L. acuminata was (1 7.41 % of control). A significant (p<0.05) decrease was observed in protein, amino acid, DNA and RNA level in the gonadal tissue of snail L. acuminata exposed to 20% and 60% of 24h LC50 of A. precatorius after exposure 24h and 96h.

DISCUSSION
It is evident from the results section that the sublethal exposure to 20% and 60% of 24h LC50 of permethrin, cypermethrin, A. sativum and A. precatorius caused a significantly change in certain biochemical parameters in the gonadal tissue of snail L. acuminata. It indicates that the biochemical changes in the gonadal tissue are altering the reproduction behaviour of these snails. Reduction of protein levels may be due to direct interference of the permethrin, cypermethrin and plant extract molluscicides with the protein synthesis. The synthesis of protein in any of a tissue can be affect in two ways by a chemical, (i) it either affect the RNA synthesis at the transcription stage or (ii) it some how affects the uptake of amino acid in the polypeptide chain. In this case the RNA synthesis would be inhibited resulting in reduced RNA as well as protein content and only the protein content would be affected [1 3, 1 4]. Amino acid levels in the gonadal tissue of the snail exposed to different preparation was significantly lower than control. It indicates that they also interfere with the biosynthesis of amino acid in the cell [1 5]. Singh et al., [1 4] have been reported that toxic effect of deltamethrin on the levels of biochemical changes in the snail Lymnaea acuminata. The effect of pyrethroids and fenvalerate caused a significant

inhibition in the protein level and enhancement in the amino acid level in the foot and hepatopancrease of snail L. acuminata [1 6]. The result section has been demonstrated that the following treatments with permethrin and cypermethrin in levels of DNA and RNA decrease in a time and dose dependent manner. The effect generally was more pronounced in the gonadal tissue. Thus the treatments with 60% of 24h LC50 of permethrin and cypermethrin the both DNA and RNA levels decreased. The decrease in DNA and RNA content in L. acuminata may be because of cell death caused by release to toxic aldehydes resulting from peroxidation [1 7]. With the reduction in DNA levels change in RNA level is certainly not surprising. With the reduced RNA level there is bound to be a fall in protein levels because of reduction in the synthesis of new protein. In conclusion, it can be stated that the component found in the synthetic (permethrin and cypermethrin) and extracts of plant alter the certain biochemical changes in snail L. acuminata.

TRANSPARENCY DECLARATION
The author declares no conflicts of interest.

REFERENCES
1 . Mas-Coma S, Bargues MD, Valero MA. Fasciolosis and other plant-borne trematode zoonosis. Int J Parasitol. 2005; 35: 1 255-1 278. 2. Mas-Coma S, Valero MA, Bargues MD. Fasciola, Lymnaeids and human fascioliasis with a global overview on disease transmission, Epidemiology, evolutionary genetics. Molecular epidemiology and control. In: D. Rollinson, S. Iain Hay (Eds.). Advances in Parasitology, Burlington: Academic Press. 2009; 69: 1 41 -1 46. 3. Agarwal RA, Singh DK. Harmful gastropods and their control. Acta Hydrochim Hydrobiol. 1 988; 1 6: 11 31 38. 4. Torgerson P, Claxton J. In: Dalton JP. (Ed.), Epidemiology and Control. CAB International, Oxon. 1 999, pp. 11 3-1 49. 5. W.H.O. Report of the WHO informal meeting on use of trichlorobendazole in fasciolosis control. 1 7-1 8 October, Geneva, Switzerland, 2006. WHO. 6. Robinson MW, Dalton JP. Zoonotic helminth infections with particular emphasis on fascioliasis and trematodiases. Phil Trans R Soc. 2009; 364-376. 7. Singh A, Singh DK, Mishra TN, Agarwal RA. Molluscicide of plant origin. Biol Agric Hortic. 1 996;

Journal of Biology and Earth Sciences, 201 3, Vol 3, Issue 2, B1 64-B1 68

B1 67

Singh and Tiwari Influence of synthetic and plant extracts on gonadal tissue of Lymnaea acuminata
1 3: 205-252. 8. Singh V, Tiwari JN. Effects of synthetic and extracts of plant on reproduction of snail Lymnaea acuminata. Res J Pharmac Biol Chem Sci. 201 2; 3(4): 705-71 0. 9. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. Protein measurement with folin phenol reagent. J Biol Chem. 1 951 ; 1 93: 265-275. 1 0. Spies JR. Colorimetric procedure for amino acid. In: S.P. Colowick, N.O. Kaplan (eds.) Methods in enzymology. Acad Press, New York. 1 957, pp: 464. 11 . Schneider WC. Determination of nucleic acid in the tissue by pentose analysis. In: Colowick SP, Kaplan NO, editors. Methods in enzymology. 1 4 New York: Acad. Press 1 957; 680. 1 2. Sokal RR, Rohlf FJ. Introduction to biostatistics. WH Freeman, San Francisco. 1 973. 1 3. Tariq M, Hussain SJ, Asif M, Jahan M. Protective effect of fruit extracts of Emblica officinalis (Gaertn) Terminalia bellerica (Roxb.) in experimental myocardial necrosis in rats. Ind J Exp Biol. 1 977; 1 5: 485-486. 1 4. Singh RN, Kumar P, Singh VK, Singh DK. Toxic effect on deltamethrin on the levels of biochemical changes in the snail Lymnaea acuminata. J Pharm Res. 201 0; 3(8): 1 739-1 742. 1 5. Singh S, Singh VK, Singh DK. Effect of molluscicidal components of Abrus precatorius, Argemone mexicana and Nerium indicum on the certain biochemical parameters of Lymnaea acuminata. Phytother Res. 1 999; 1 3: 21 0-21 3. 1 6. Singh A, Singh DK, Agarwal RA. Effect of cypermethrin mexacarbate and phorate on phospholipids and lipid peroxidation in snail Lymnaea acuminata. Bull Environ Contam Toxicol. 1 993; 51 : 68-71 . 1 7. Singh RN, Kumar P, Singh VK, Singh DK. Effect of binary combinations of deltamethrin + MGK 264 on the level of phospholipid and lipid peroxidation in the snail Lymnaea acuminata. Chemosphere. 2008; 73: 1 032-1 035.

Journal of Biology and Earth Sciences, 201 3, Vol 3, Issue 2, B1 64-B1 68

B1 68