Effect of two seaweeds Ulva fasciata and Grateloupia lithophila as larvicide against mosquito vector Aedes aegypti

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1 INT J CURR SCI 2013, 6: E RESEARCH ARTICLE ISSN Effect of two seaweeds Ulva fasciata and Grateloupia lithophila as larvicide against mosquito vector Aedes aegypti Josmin Laali Nisha L.L and Poonguzhali T.V P.G & Research Department of Botany, Queeen Mary s College, Chennai , India Corresponding author: pooqmc@gamil.com, josminll@rediffmail.com; Phone: Abstract The uses of insecticide for mosquito control are increasingly becoming limited. The control measure were largely based on the use of chemical pesticides which develop resistance in the target population The problems in continued long term use of toxic insecticide include pest resistance and negative impact on natural enemies, which suggest a clear need for alternatives. Biolarvicides are biological products that are added to water to kill mosquitoes at the larval stage. Biological control is the most effective way to fight against larva as it prevents the birth of new mosquitoes and does not pollute the environment or pose health risks. Aedes aegypti, is responsible for dengue fever and yellow fever, in India where the number of these fever cases has increased significantly in recent years. Seaweeds have shown a wide range of bioactive properties. In this connection, the present study was an attempt to find out the larvicidal activity of different solvent extracts of two seaweeds Ulva fasciata and Grateloupia lithophila against the dengue and yellow fever causing mosquito, Aedes aegypti. Keywords: Ulva fasciata, Grateloupia lithophila, larvicide, seaweeds, pesticides Received: 20 th Nov 2012; Revised: 06 th January; Accepted: 14 th February; IJCS New Liberty Group 2013 Introduction The attached algae that live in the sea or the benthic marine algae is known as, seaweeds. For more than a thousand years we humans have found uses for seaweed. Mehlhorn, 2006a, b; Rahuman et al., 2008a). Many studies on plant extracts against insect larvae have been conducted around the world. The change in enzyme system may develop resistance for mosquito, which resulting in more The potential uses of seaweeds have been have been rapid detoxification or sequestration of the insecticide, or organised into the following categories: i) aesthetics, ii) agriculture, horticulture and agronomy, iii) uses in animal aquaculture, iv) cosmetics, v) environmental health, monitoring and remediation, vi) food, vii) health, thalassotherapy and wellness, viii) industry, ix) pharmaceutical and pharmacology, x) science, technology & biomedicine, xi) miscellaneous uses. But there are also a myriad of human uses for algae. Most researchers have reported the bioactivity of extracts from various plants against agricultural pest and medically important vectors (Morimoto et al., 2002; Koul et al., 2003; Amer and due to mutations in the target site preventing the insecticide target site interaction (Hemingway et al., 2004). The uses of insecticide for mosquito control are increasingly becoming limited. C. tritaeniorhynchus was found susceptible to permethrin and resistant to DDT, dieldrin, fenitrothion and propoxur (Bansal and Singh, 1995) and resistant to organophosphorous insecticides (Watanabe et al., 1991). The control measure were largely based on the use of chemical pesticides, including chlorinated hydrocarbons, organophosphates, and

2 carbamates (Shetlar, 2001), which develop resistance in the target population (Hollingsworth et al., 1994; Han and Li, 2004). The problems in continued long term use of toxic insecticide include pest resistance and negative impact on natural enemies, which suggest a clear need for alternatives. Biolarvicides are biological products that are added to water to kill mosquitoes at the larval stage. Biological control is the most effective way to fight malaria as it prevents the birth of new mosquitoes and does not pollute the environment or pose health risks. Aedes aegypti, is responsible for dengue fever and yellow fever, in India where the number of these fever cases has increased significantly in recent years. Seaweeds have shown a wide range of bioactive properties (Barbosa et al., 2007; Wang et al., 2008; Bazes et al., 2009; Sabina et al., 2009; Ravikumar et al., 2009 and 2011). In this connection, 35 C in a shaker at 120 rpm for 7 days for the extraction of active ingredients. From this stock solution dilutions were made to prepare different concentrations, such as 100, 200, 300, 400 and 500 mg/l respectively, including positive (with 2% methanol, acetone and benzene) and negative controls (larvae exposed to dechlorinated water without methanol, acetone and benzene). Test mosquito larvae Larvae of Aedes aegypti were collected from clean stagnant water areas of Chennai. It was maintained at 27 ± 2 C, 75 85% relative humidity. The larvae were fed with dog biscuits and yeast at 3:1 ratio. Bioassay The larvicidal bioassay followed the World Health Organization (WHO) standard protocols World Health Organization (1981). Instructions for determining the the present study was an attempt to find out the larvicidal susceptibility or resistance of mosquito larvae to activity of different solvent extracts of two seaweeds Ulva fasciata and Grateloupia lithophila against the dengue and insecticides was carried out on the basis of WHO/VBC, (81:807) with slight modifications. Bioassays were yellow fever causing mosquito, Aedes aegypti. conducted with larvae, which was collected using a Materials and Methods Collection of algae and extract preparation Two seaweed samples, Ulva fasciata Delile and Grateloupia lithophila Boergesen were collected from the Coast of Kovalam, near Chennai. Healthy algal material were harvested manually and washed thoroughly in running water to remove epizoones, epiphytes, animal castings, sand, calcareous and other adhering detritus matters. Cleaned algal materials were shade dried under Pasteur pipette, placed on filter paper for removal of excess water and transferred (25 per test) with a tiny brush into beakers containing different concentrations of algal extracts (100, 200, 300, 400 and 500 mg/l) with 1000 ml of tap water each. Larvae were exposed to the samples at room temperature for 24 hours and the mortality/survival were registered. Each test was run in triplicate. The persistence of larvicidal activity of the algal extract was tested by running bioassays with the same samples after room temperature for 4-5 days. The completely dried 15, 30 and 60 days. material was powdered using electric blender. Three different extracts (methanol, acetone and benzene) were prepared by submerging the powder in three different flask of each containing 1000 mg/l and placed at Statistical analysis The larval mortality in each concentration and control was recorded after 24 hrs of exposure. Percentage mortalities were corrected for the natural mortality

3 observed in the negative controls using Abbots (1925) the three different extracts of G.lithophila at different formulae; concentrations ( mg/l) showed that acetone P = PI C / 1 C where PI denotes the observed mortality rate and C means the natural mortality. The median lethal concentration or dose (LC50 and LD90) was calculated using Probit analysis (Finney 1971) that has been recommended by OECD guideline (Poonguzhali and Nisha, 2012). Results Results of the larvicidal activity of three different extracts (methanol, acetone, and benzene) of U. fasciata and G.lithophila against the larvae of A. aegypti was performed under laboratory evaluation. In terms of lethal concentration for 50% and 90% mortality (LC 50 and LC 90 ) values were represented as follows: LC 50 value of the methanol extract of U. fasciata was , followed by benzene extract and acetone extract was while the LC 50 value of the acetone extract of G. lithophila was followed by benzene extract and methanol extract LC 90 value of the benzene extract of U. fasciata was followed by acetone extract and methanol extract; while for G. lithophila LC 90 value of the methanol extract was , acetone extract was and benzene extract was (Table 1). It illustrates the larval mortality rate of A. aegypti after the treatment of the three different extracts of U.fasciata at different concentrations ( mg/l). The LC 50 values of U. fasciata revealed that the larvae A. aegypti was more susceptible to methanol extract followed by benzene and acetone (Methanol> Benzene > Acetone) and the LC 90 value showed that the larva was more susceptible to benzene extract followed by acetone and methanol extract. The larval mortality rate of A. aegypti after the treatment of extract was more efficient than the other extracts and the lethal hierarchy is (Acetone > Benzene > Methanol) (Table 2). Table 1. Effect of methanolic, acetone and benzene extracts of U. fasciata against A. aegypti mosquito larvae Extract Methanol Acetone Benzene LC50 (mg/l) % LCL UCL LC90 (mg/l) % LCL UCL Table 2. Effect of methanolic, acetone and benzene extracts of G. Lithophila against A. aegypti mosquito larvae Extract Methanol Acetone Benzene LC50 (mg/l) % LCL UCL LC90 (mg/l) % LCL UCL LC50 and LC90 = [Lethal Concentration] Concentration required to kill 50 and 90% of the test populations respectively; LCL: Lower Limit; UCL: Upper Limit. Discussion Seaweeds have been found to be vital source of useful bioactive substance since two decades ago. There is no previous report on the mosquito larvicidal activity of U. fasciata and G. lithophila from the Kovalam coast of Tamil Nadu. Of the two algae screened G. lithophila was found to be effective against A. aegypti larva in all the 3 extracts. Among the seaweeds G. lithophila was better than U.fasciata which could be revealed by the lower LC 50 values exhibited by G. lithophila against all the three

4 extracts. The organic extract of U. fasciata found to be larvicidal in the previous reports were less active in the present study. This may be due to the fact that the bioactivity of same taxa can vary with geographical zone and could depend on ecological parameters such as irradiance and nutrients (Marti et al., 2004). Osman et al. (2010) demonstrated the post coital contraceptive activity from the the crude extract in marine red algae Gelidiella acerosa is due to the presence of various phytochemical components such as alkaloids, phenols, flavonoids, steroids tannins and carbohydrates. The acetone extract of seaweeds Caulerpa scalpelliformis and Dictyota dichotoma exhibited mosquito larvicidal activity against Aedes aegypti (Thangam and Kathiresan, 1991). Albeit the larvicidal potential of seaweeds was well established, no evidence was available on the field trials of seaweed based mosquito repellents. Mullai and Jebanesan (2006) reported the larvicidal efficacy of the leaf extract of Cucumis pubescens with four different solvents against late third instar larvae of Anopheles stephensi, Culex quinquefasciatus and A. aegypti. The seaweeds (U.fasciata and H. musciformis) produced 100% larva mortality at 10 mg/ml (Selvin and Lipton, 2004). Conclusion India has rich reserve of coastal wealth. Hence, the seaweeds which were collected from the east coast of India have been collected and then its potential for mosquito larvicidal activities can be commercially used for eco friendly mosquito control programme. This will be an alternate source of income for the fisher men involved in algal collection. References Abbott WS (1925). A meth od of co mputin g th e effectiveness of insecticides. Journal of Economic Entomology, 18: Amer A, Mehlhorn H (2006a). Larvicidal effects of various essential oils against Aedes, Anopheles and Culex larvae (Diptera, Culicidae). Parasitol Res., 99: Amer A, Mehlhorn H (2006b). Persistency of larvicidal effects of plant oil extracts under different storage conditions. Parasitol Res., 99: Bansal SK, Singh KV (1995). Susceptibility status of two species of Japanese encephalitis vectors to insecticides in the Thar desert, District of Bikaner (Rajasthan). Indian J Med Res.,101: Barbosa JP, Fleury BG, da Gama BAP, Teixeira VL, Pereira RC (2007). Natural products as antifoulants in the Brazilian brown alga Dictyota pfaffii (Phaeophyta, Dictyotales). Biochem Syst Ecol., 35: Finney DJ. (1971). Probit analysis. 3 rd edition. Cambridge University Press, Cambridge (England). Han Z, Li F (2004). Mutations in acetyl-cholinesterase associated with insecticide resistance in the cotton aphid, Aphis gossypii Glover. Insect Biochem Mol. Biol., 34: Hemingway J, Hawkes NJ, McCarroll L, Ranson H (2004). The molecular basis of insecticide resistance in mosquitoes. Insect Biochem. Mol. Biol., 34: Hollingsworth RG, Tabashnik BE, Ullman DE, Johnson MW, Messing R (1994). Resistance of Aphis gossypii (Homoptera: Aphididae) to insecticides in Hawaii: spatial patterns and relationship to insecticide use. J. Econ. Entomol., 87:

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