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Larvicidal Activities of Different Parts of Melia Azedarach Linn. Against Culex Quinquefasciatus Say

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Larvicidal Activities of Different Parts of Melia Azedarach Linn. Against Culex Quinquefasciatus Say Journal of Basic & Applied Sciences, 2012, 8, 23-28 23 Larvicidal Activities of Different Parts of Melia azedarach Linn. against Culex quinquefasciatus Say. (Diptera: Culicidae) Ikram Ilahi*,1, Imran Khan1, Mohammad Tariq1 and Izhar Ahmad2 1Departmrnt of Biotechnology, University of Malakand, Chakadara, Dir (Lower), Khyber Pakhtunkhwa, Pakistan 2Department of Plant Sciences, Faculty of Biological sciences, Quaid-i-Azam University, Islamabad, Pakistan Abstract: Melia azedarach is a perennial tree, belongs to the family Meliaceae, and locally known as Tora Shandi in Swat and Dir regions of Khyber Pukhtunkhwa (K.P.K). The present study was aimed to evaluate the larvicidal activity of aqueous extracts of different parts of Melia azedrach against Culex quinquefasciatus. Various concentrations (50, 100, 500, 1000, 1500 and 2000 ppm) of aqueous extracts of leaves, fruits and bark of Melia azedarach were tested for larvicidal activity against Culex quinquefasciatus. There occurred a continuous increase in mortality of 3rd and 4th instar larvae with increase in concentration of the extracts. At 50ppm, the fruit extract caused 1.6 + 2.2 % mortality, leaves extract 17.60 + 6.0 % and bark extract caused 17.60 + 7.3 % mortality. The LC50 value for fruits extract was 2035.13ppm, leaves extract 612.250ppm and for bark extract 368.3ppm. Among the aqueous extracts of fruits, leaves and bark, the aqueous extract of bark was proved to be more toxic and an efficient larvicidal against Culex quinquefasciatus. The findings of the present study indicate that the aqueous extract of the bark of Melia azedarach of Dir region of khyber pakhtunkhwa can serve as a good source of preparations for pest control especially mosquito control. Keywords: Aqueous extracts, Culex quinquefasciatus, Mortality, LC50. INTRODUCTION The members of Meliaceae plant family are known to contain a variety of compounds, which show Melia azedarach Linn. is one of the medicinal plants insecticidal, antifeedant, growth regulating and commonly known as "Persian Lilac", "Bakain" or development modifying properties [12, 13]. Fruit "think", grows wild throughout the Sub-Himalayan belt. extracts of Melia azedarach and Azadirachta indica It is cultivated in India and Pakistan for both ornamental elicit a variety of effects in insects such as antifeedant, and medicinal purposes [1]. It is a perennial tree and growth retardation, reduced fecundity, moulting belongs to the family Meliaceae, its English name is disorders, morphogenetic defects, and changes of china berry and locally known as Tora Shandi in Swat behavior [7, 14-17]. Many plants of Meliaceae family and Dir regions of Khyber Pakhtunkhwa (K.P.K). possess insecticidal properties against fecundity and fertility of mosquito vector and are at the same time Various preparations of Melia azedarach are being very eco-friendly [18-19]. used for the treatment of several diseases [1, 2]. There are several reports on the analgesic, anticancer, Control of mosquito is essential as many species of antiviral, anti malarial, antibacterial, antifungal and anti- mosquitoes serve as vectors for spreading of malaria, fertility activity of this plant [3]. The use of M.azedarach filariasis, dengue and many other viral diseases and in stomach ache, intestinal disorders, uterine illnesses, they cause unbearable biting irritations [20-22]. A more cystitis, diuretic and febrifuge has been reported [4]. efficient and attractive approach in mosquito control The powder of the dried fruits of this plant is claimed to programs is to target the larval stage in their breeding be an effective therapy for the treatment of diabetes [5]. sites with larvicides [23-24]. Various organophosphates In the field of veterinary medicine, the ovicidal and (Temiphos and Fenthion) and insect growth regulators larvicidal activity of M. azedarach extracts on the (Diflubenzuron and Methoprene) are widely used as helmenthus Haemonchus contortus has been reported larvicides for the control of mosquitoes [25]. These [6]. This plant has also been studied for its antimalaria synthetic pesticides have caused the development of [7, 8] as well as spermicidal [9]) and antifungal activity resistant insect strains, ecological imbalance and harm [10]. to mammals including humans [26]. The best alternative for avoiding these problems could be the The family Meliaceae to which Melia azedarach use of plant-based insecticides, which are quickly belongs contains 45 genus and over 750 species [11]. degradable in the environment and their source is renewable [27]. *Address corresponding to this author at the Department of Biotechnology, University of Malakand, Chakdara, Dir (L) Khyber Pakhtunkhwa, Pakistan; Tel: +923018539845; Fax: +92945763491; E-mail: [email protected] ISSN: 1814-8085 / E-ISSN: 1927-5129/12 © 2012 Lifescience Global 24 Journal of Basic & Applied Sciences, 2012 Vol. 8, No. 1 Ilahi et al. In the present study Melia azedarach Linn. was adults were placed in test tubes containing silica gel. In investigated for its larvicidal activity against mosquito addition to the study of the adults, permanent slides of Culex quinquefasciatus Say. Culex quinquefasciatus 4th instar larvae were also prepared. The adults and is a vector of widely distributed tropical disease larvae were studied for taxonomic characters by using “lymphatic filariasis” [28] and also causes unbearable binocular microscope and identification was made up to biting irritations. The insecticidal efficacy of extracts of species level with the help of taxonomic keys provided M. azedarach has been evaluated against ticks [29], in the literatures [33-34]. All the specimens were Ae. aegypti [8, 30] and An.stephensi [31]. There are belonging to the species Culex quinquefasciatus. So it also some reports on the evaluation of larvicidal activity was confirmed that the ditch harbors only Culex of Melia azedarach against Culex quinquefasciatus quinquefasciatus. [32]. Larvicidal Bioassay The aim of the present study was to evaluate the larvicidal activity of aqueous extracts of different parts For conduction of larvicidal activity, larvae were of Melia azedrach against Culex quinquefasciatus. collected from the same site in a large plastic jar and brought to the laboratory. A stock solution of MATERIALS AND METHODS 10,000ppm of each extract was prepared in distilled water. From the stock solution of each extract, 100 ml Plant Material each of 2000, 1500, 1000, 500, 100 and 50 ppm were prepared in different 500ml plastic cups. The cups were The fruits, leaves and bark of Melia azedarach Linn. labeled accordingly. For bioassay of each extract, 25 were collected from the campus of University of rd th larvae of late 3 and early 4 instar were transferred to Malakand, Khyber Pakhtunkhwa, Pakistan during June, each plastic cup with the help of a large mouthed glass 2011. The plant was identified by Mr. Ilyas Iqbal, dropper. Larvae were fed with a diet of finely ground Assistant Professor, Departmenty of Botany, University brewer’s yeast and dog biscuits in 3:2. A control was of Malakand. A voucher specimen was deposited in the also maintained by adding only larval food (yeast and Herbarium of University of Malakand. dog biscuit in 3:2). All the experiments were carried out Extraction at 30 + 2°C and 70-75 relative humidity. The numbers of dead larvae were counted after 24 h of exposure. Fruits, leaves and bark of Melia azedarach were Dead larvae were identified when they failed to move cleaned and shade dried. One kilogram each of fruits, after prodding with a needle in the cervical or siphon leaves and bark were coarsely ground and then soaked region. Five replicates of the experiment were made. in 3000 ml of distilled water for 7 days with occasional After recording the number of dead larvae, all the shaking. Each soaked material was filtered through a larvae (dead and live if present) were identified to muslin cloth, each filtrate was placed in separate confirm the species. beaker. The filtrates were dried under reduced Statistical Analysis pressure using rotary evaporator (Heidolph Laborta 4000 efficient). The dried crude extracts were stored in The dose-response data was subjected to Linear refrigerator at 4ºC. regression analysis to point out the relationship Test Organism between the increase in extract concentration and larval mortality. For the calculation of LC50 values the Before the conduction of larvicidal bioassay, 100 data was subjected to probit regression analysis. To samples of 3rd and 4th instars larvae of mosquitoes compare the aqueous extracts of fruits, leaves and were collected with the help of a glass dropper from a bark, the data was subjected to Duncan Test of Post ditch of stagnant water at the campus of University of Hoc Multiple Comparisons in One Way Anova. For all Malakand. The immature stages were reared in plastic these analysis computer software SPSS 16.0 was jars. To each jar 200 ml of water was added from the used. collection spot to ensure food supply. The jar openings RESULTS were closed with net cloth. The larvae were reared to adulthood in the laboratory. The adult mosquitoes were The larvicdal activities of various concentrations of captured with the help of aspirator and killed with a aqueous extracts of different parts of Melia azedarach cotton swab of chloroform in an airtight container. The Larvicidal Activities of Different Parts of Melia azedarach Linn. Journal of Basic & Applied Sciences, 2012 Vol. 8, No. 1 25 were studied against Culex quinquefasciatus. There value for fruit extract was 2035.13ppm. The lowest occurred a continuous increase in mortality of 3rd and concentration (50ppm) of leaf extract caused 17.60 + 4th instar larvae with increase in concentration of the 6.0 % mortality and the highest concentration caused extracts. The mortality caused by each of the fruits, 81.60 + 4.6 % mortality. The LC50 value for leaves leaves and bark aqueous extract was subjected to extract was 612.250ppm. The lowest concentration linear regression analysis. In each case the value of R (50ppm) of bark extract caused 17.60 + 7.3 % mortality square was about 0.9 at P = 0.05 significance (Table and the highest concentration caused 88.0 + 6.3 % 1).
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