LARVICIDAL EFFICACY OF PIPER LONGUM FRUIT EXTRACT AGAINST AEDES AEGYPTI, A DENGUE FEVER VECTOR

VISTAS Vol. 4, No. 1, 2015, pp. 123-127 ISSN: 2319-5770, e-issn 2394-1138 LARVICIDAL EFFICACY OF PIPER LONGUM FRUIT EXTRACT AGAINST AEDES AEGYPTI, A DENGUE FEVER VECTOR Vinu Rajan P.K. 1, Sreedev P. 2, Misvar Ali K. 3 and Aneesh E.M. 4 1,3Research and Development, Bharathiar University, Coimbatore 2,4Communicable Disease Research Laboratory, St. Joseph s College, Irinjalakuda 4Corresponding author, Department of Zoology, St. Joseph s College, Irinjalakuda Abstract The larvicidal efficacy of hexane, ethyl acetate, acetone and methanol fruit extracts of Piper longum against the early fourth instar larvae of Aedes aegypti was studied. Insecticidal susceptibility tests were carried out employing WHO standard method and the mortality was observed after 24 h exposure. The methanol leaf extract was found to be the effective larvicide with LC 50 value of 221.7 mg/l after 24 h. The result indicates promising larvicidal activity of Piper longum against Aedes aegypti. Further investigations are needed to isolate and identify the active principles involved, level of toxicity and their mode of action. Plants are chemical factories of nature. Human have used plant parts, products and metabolites in pest control since early historical times. Some plant metabolites have strong medicinal and insecticidal property; by using plant extracts in recent times man has been able to control certain pests and vectors. Some important phytochemical products pyrethrum, derris, quassia, nicotine, hellebore, anabasine, azadirachtin, dlimonene, camphor, and terpenes that have been used as insecticides (Wood, 2003). Plants and plant based insecticides were widely used throughout the world, developed countries using plant origin insecticides more than organic insecticides (Casida and Quistad, 1998). Mosquitoes are etiological agents for some of the devastating diseases like malaria, filariasis, chikunguniya, dengue etc. Mosquitoes serve as vector for various tropical and subtropical diseases which cause destructive effects to human (Kovendan and Murugan, 2011). They do not only transmit parasites and pathogens but they also source of allergic reaction that includes local skin and systemic sensitivity (Cheng et al, 2003). Dengue is one of the most significant viral diseases transmitted by Aedes aegypti because it afflicts humans worldwide whose symptoms ranging from mild fever to a severe and potentially life threatening hemorrhagic disease. Aedes aegypti is of supreme concern because of its wide distribution and close association with humans (Ravikumar et al, 2011). World Health Organization (WHO) stated that about 2/5 of the global human population are currently under threat of dengue. Due to the pathogenic diseases and serious harms caused by mosquitoes, controlling them has been the primary subject of several new researches over the past few years (Invest and Lucas, 2008). The systemic application of insecticides is a common and widely accepted approach to control mosquito population, it s a very useful method but also have draw backs. It will produce resistant mosquito colonies and causes disease outbreak (Liu et al., 2006). Chemical insecticides are not good in the environmental point of view, majority of the applied insecticide contaminate the environment and reaches non target organisms (Arufe et al., 2007). The negative impact of these chemical http://www.groupexcelindia.com/vistas/index.html

124 ISSN: 2319-5770 e-issn 2394-1138 insecticides shifted the research towards the development of naturalistic agents. Nowadays science is intensified towards plants and the plant kingdom is receiving renewed attention as mosquitocides. The plant extracts have the property to kill the mosquito larvae without affecting the other organism and environment (Heldin et al 2007, Arnason et al 1989). Sukumar et al published various reviews on the action of phytochemicals against mosquitoes (Sukumar et al, 1991), Park et al. Carried out a detailed laboratory study on extracts of fruits of Piper nigrum L. (Piperaceae) against larvae of Culex pipiens L., Aedes aegypti (L.), and Ae. togoi Theobald (Park et al. 2002). Mulla and Su reviewed neem products with activity and bio-efficacy against arthropods of medical and veterinary importance. They noted that a variety of neem components exhibited effects such as antifeedancy, growth regulation, ovicidal effects, reproduction suppression, and changes in biological fitness in many species of arthropods of medical and veterinary importance (Mulla and Su, 1999). Madhu et al reported the insecticidal activity of piper longum against Culex. quinquefasciatus ( Madhu et al, 2011). Piper longum (Indian long pepper,thippali or Pippali), belonging to the family of piperaceae is a slender aromatic climber with red fruits. These fruits are greyish black with a pungent smell when dried and reported as a good remedy for the treatment of gonorrhoea,menstrual pain,tuberculosis,sleeping problems, resipatory track infections and arthritic condition (Nadkarni 2007). Our study is intended to bring out the larvicidal efficacy of fruits of Piper longum against Aedes aegypti. MATERIALS AND METHOD Fruits of Piper longum were purchased from local markets of Thrissur. All the organic solvents used in these experiments were of analytical grade and purchased from Merck chemicals India, Aedes aegypti larvae used in the experiments were maintained in Communicable Disease Research Laboratory, St.Joseph s College, Irinjalakuda. PHYTO EXTRACTION A soxhlet extraction was carried out with fruits using a series of organic solvents of increasing polarity viz, Petroleum ether, hexane, acetone and methanol until exhaustion. Solvents were distilled in a vacuum rotary evaporator under reduced pressure of 20-22 mmhg at 35 0 c and extract concentrates were further evaporated to complete dryness at room temperature. Surface tension of each test solution was measured using stalagmometer to ensure the complete miscibility. Each extracts were assayed for their larvicidal efficacy with variable concentrations and doses were fixed to give larval morality ranging from 10 to 98 percent. As petroleum ether extract was found to be the most bioactive, it was selected for further analysis. LARVAL BIOASSAY Larval susceptibility tests of different solvent extracts and isolated compound were performed employing WHO standard procedure. Various concentrations of the extracts were prepared by serial dilutions of a stock solution in acetone. Batches of 25 early fourth instar larvae were released into glass beakers of 500 ml capacity containing 249 ml of dechlorinated tap water and 1.0 ml of extract. Control beakers contained 25 test organisms and 249 ml of tap water along with 1.0 ml acetone. Treated and control beakers were maintained at same conditions (25±20 C), 12 h

VISTAS Vol. 4, No. 1 125 light/dark regime. No food was provided to the larvae during the test period of 24 h till the morality was monitored. All treatments were replicated four times. The larvae were considered as dead or moribund, if they were not responsive to gentle prodding with a fine needle. The results were expressed as percent mortality. RESULTS The results for the toxicity of the Piper longum fruit extracts by different solvents tested against Aedes aegypti larvae are presented in Table 1. The fruit extracts of Piper longum exhibit mortality against the fourth instar larvae of Aedes aegypti. Methanol extract has exhibited minimum LC 50 and LC 90 of 221.7 ppm and 718.1 ppm respectively. It was followed by the hexane extract by producing 50% mortality with 318.95 ppm. The LC 50 value of ethyl acetate extract was 489.81ppm with LC 90 of 2252.99. The least larvicidal activity was detected in acetone extract of Piper longum with LC 50 of 611.96 ppm. Extraction solvent Hexane Ethyl acetate Acetone Methanol Table 1: Efficacy of Different Solvent Extracts of Piper Longum Fruit against Fourth instar Larvae of Aedes Aegypti DISCUSSION 09.8±3.6 28.6±2.0 42.5±2.0 63.6 ± 1.5 77.3 ± 2.5 15.9±0.5 35.6±1.0 53.6±1.5 74.2±2.0 94.3±1.5 19.8±1.1 46.1±1.0 65.1±2.5 83.7±1.6.0±0 2.00±0.19 489.81 2.28±0.34 611.96 2.50±0.40 221.7 411.00-545.73 205.00-479.18 140.59-369.95 2252.99 1194.31 718.1 Conc. in %mortality Slope±SE LC50(mg/L) (95%FL) LC90(mg/L) (95%FL) χ 2 (df) Regression ppm equation 14.2 ±2.0 26.5 ±1.1 275.03-1112.50-37.3 ±2.1 1.78±0.40 318.95 928.40 14.00(3) Y = 1.7853 1425.95 X +.1631 52.0 ±0.5 82.0 ±3.6 1551.33-3051.53 735.50-3963.06 524.19-2420.99 0.90(3) Y = 2.0067 X + -.3656 9.62(3) Y = 2.2831 X + -.7440 11.68(3) Y = 2.5010 X + -1.0048 A number of researches on the field of vector control have revealed the efficacy of different phytochemicals obtained from various plants against different species of mosquitoes. Botanical derivatives are considered to be ecofriendly and can be used in mosquito larval control ensuring maximum safety in any ecological condition. The biological activity of plant extracts must be due to diverse compounds including phenolics, terpenoids and alkaloids present in plants (Rafael et al., 2009). The present study was aimed to utilize the fruits of Piper longum as mosquito control agent. Our investigations revealed that methanol extracts of Piper longum exhibited 2.03 fold more larvicidal potential than acetone extracts against early fourth instars of Aedes aegypti.

126 ISSN: 2319-5770 e-issn 2394-1138 The type of solvents selected affects extract efficacy between different phytochemicals of varying volatility is present in the final extraction (Shaalan et al., 2005). The current study tested the larvicidal efficacy of extracts employing solvents of increasing polarity from hexane (non polar, p=0.1) to water (the most polar one with a polarity index (p) of 10.2). Data on Piper longum extract showed an equivalent relationship exists between extract efficacy and solvent polarity where efficacy increases with increasing polarity. This is not in line with the observation made by Aivazi and Vijayan(2009) in oak gall extracts and Prathibha and Vijayan(2010) in Euodia ridleyi leaf extract as where they got converse relationship of increasing efficacy with decreasing polarity. CONCLUSION The first hand information of the current research clearly shows the potential of Piper longum as a possible larvicidal agent against Aedes aegypti. The perusal of literature on phytochemical assays and the present study on the target species reveal that botanical derivatives can be explored as a powerful weapon in preventing deadly transmission of vector borne diseases. The effective larvicide obtained from the current study with LC 50 value of 221.7 mg/l after 24 h indicates promising larvicidal activity of Piper longum against Aedes aegypti Further study is required to reveal the active principle involved, level of toxicity and their mode of action. ACKNOWLEDGEMENT The authors are thankful to the Principals of St. Joseph's College, Irinjalakuda and Christ College, Irinjalakuda for the facilities provided and KSCSTE for the financial assistance under SRSLS Scheme. REFERENCES Aivazi, A.A. and Vijayan, V.A. (2009), Larvicidal Activity of Oak Quecus Infectoria Oliv. (Fagaceae) Gall Extracts Against Anopheles Stephensi, Liston J. Parasitol. Res., Vol. 104: pp. 1289 1293. Arnason, J., Philogene, B. and Morand, P. (1989), Insecticides of Plant Origin, American Chemical Society Journal, Vol. 387, p. 213. Casida, J.E. and Quistad, G.B. (1998), Golden Age of Insecticide Research: Past, Present or Future? Annu. Rev. Entomol, Vol. 43: pp. 1 16. Cheng, S.S., Chang, H.T., Chang, S.T., Tsai, K.H. and Chen, W.J., (2003), Bioactivity of Selected Plant Essential Oils Against the Yellow Fever Mosquito Aedes Aegypti Larvae, Bioresource Technol, Vol. 89, pp. 99 102. Hedlin, P.A., Holingworth, R.M., Masler, E.P., Miyamoto, J. and Thopson, D.G., (1997), Phytochemicals for Pests Control, American Chemical Society, p. 372. Invest, J.F. and Lucas, J.R., (2008), Pyroproxyfen as a Mosquito Larvicide, Proceedings of the Sixth International Conference on Urban Pests. Jaswanth, A., Ramanathan, P. and Ruckmani, K., (2002), Evaluation of the Mosquitocidal Activity of Annona Squamosa Leaves Against Filarial Vector Mosquito, Culex quinquefasciatus Say. Indian J Exptl Biol, Vol. 40: pp. 363 5. Kovendan, K. and Murugan, K. (2011), Effective of Medicinal Plants on the Mosquito Vectors from the Different Agroclimatic Regions of Tamil Nadu, India, Advances in Environmental Biology, Vol. 5(2), pp. 335 344. Madhu, S.K., Vijayan, V.A. and Shaukath, A.K. (2011), Bioactivity Guided Isolation of Mosquito Larvicide from Piper Longum, Asian Pacific Journal of Tropical Medicine, pp. 112 116. Mulla, M.S. and Su., T. (1999), Activity and Biological Effects of Neem Products Against Arthropods of Medical and Veterinary Importance, J. Am. Mosq. Contr. Assoc., Vol. 15: pp. 133 152.

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