Bangladesh Journal of Pharmacology Research Article Larvicidal activity of Oestrum noc- turnum on Aedes aegypti BJP Introduction Dengue fever is considered as a serious public health problem in the world, mainly in tropical countries where the favorable environmental conditions are responsible for the proliferation of vectors Aedes aegypti. Among the arbovirus in India, distribution of all the dengue virus type is continuously expanding. Remarkably the reemergence of Chikungunya virus (CHIK) since 2005 is posing an additional concurrent diseases burden in the country including the Maharashtra. Both these virus are born by the mosquito A. aegypti (L) (Diptera:Culicidae) (Fulmali et al., 2008; Kumar et al., 2008). Periodic treatment with chemical insecticides and synthetic pyrethroids are done in breeding sits. Due to environmental concern on use of existing synthetic insecticides for vector control and further risk of development of widespread insecticides resistance in disease vector; interest on possible use of environment friendly natural products such as extracts of plants or plant parts increased for vector control. Sukumar et al. (1991) listed 346 species for 276 genera and 99 families which have been tested against mosquitoes for various effects such as toxicity, growth inhibition, ovipositional determinacy and repellent. This list includes many species from Solanaceae family. Recently, Ghosh and Chandra (2006) and Ghosh et al. (2008) evaluated phytosteroidal compound of mature leaves of day jasmine Cestrum diurnum (Solanaceae:Solanales) against larvae of Culex quinque- fasciatus (Diptera:Culicidae) and Anopheles stephensi. This study with plant extracts has been pointed as a promising alternative to combat this vector. In this work we evaluate the potential of extract from C. nocturnum as larvicide. Materials and Methods Plant species C. nocturnum leaves were collected from gardens at Nashik city, Maharashtra State, India, in December 2008. Leaves were shed dried and powdered. Plant extraction The leaves were macerated and sequentially extracted with hexane, ethyl acetate and methanol using soxhlet and percolation extraction separately. The solvent were evaporated on rotary evaporator. Any extract exhibiting significant larvicide activities were further fractioned by filtration in silica get eluted with hexane-ethyl Abstract Aedes aegypti is a vector parasite of the Dengue. New method to control the population of this insect is necessary. In the present work we evaluated the potential of extract from Cestrum nocturnum as larvicide. Methanol extract outstand as highly active larvicide, achieving 100% larval mortality in 24 hours when tested in the concentration of 45 pg/mL (soxhlet) and 25 pg/mL (percolation). Any extract exhibiting significant larvicide activity was further fractioned and the fraction tested according to the WHO protocol. One fraction derived from methanol extract present remarkable LC100 at 12 pg/ mL. LC50 of methanol extract and active fraction were found 14 pg/mL and 6 pg/mL respectively. These fractions will be submitted to further fractions aiming to identify the molecules responsible for the larvicide activity. Article Info Received: 5 April 2010 Accepted: 9 May 2010 Available Online: 2 June 2010 DOI: 10.3329/bjp.v5i1.4714 Cite this article: Jawale C, Kirdak R, Dama L. Larvicid- al activity of Oestrum nocturnum on Aedes aegypti. Bangladesh J Pharma- col. 2010; 5: 39-40. Larvicidal activity of Oestrum nocturnum on Aedes aegypti Chetan Jawale1, Rambhau Kirdak2 and Laxmikant Dama3 1Department of Zoology, HPT Arts & RYK Science College, Nashik 422005, Maharashtra; 2Directorate of Higher Education, Pune, Maharashtra; 3Department of Zoology D.B.F. Dayanand College of Arts and Science, Solapur 413004, Maharashtra, India. This work is licensed under a Creative Commons Attribution 4.0 License. You are free to copy, distribute and perform the work. You must attribute the work in the manner specified by the author or licensor. A Journal of the Bangladesh Pharmacological Society (BDPS) Bangladesh J Pharmacol 2010; 5: 39-40 Journal homepage: www.banglajol.info Abstracted/indexed in Academic Search Complete, Agroforestry Abstracts, Asia Journals Online, Bangladesh Journals Online, Biological Abstracts, BIO- SIS Previews, CAB Abstracts, Current Abstracts, Directory of Open Access Journals, EMBASE/Excerpta Medica, Google Scholar, HINARI (WHO), Interna- tional Pharmaceutical Abstracts, Open J-gate, Science Citation Index Expanded and Social Sciences Citation Index ISSN: 1991-0088 acetate mixture of increasing polarity, eight fraction were obtained. Bioassay Larvae were reared (Pelah et al., 2002) and third instars larvae were selected for bioassay. Larvae were trans- ferred into the test solution with pasture pipette (20 larvae/solution). As a solvent, DMSO is used to soluble the extract in test water. Mortality of each test extract and fractions were determined after 24 hours exposure at 28°C following the protocol of WHO (1981). Mortality was corrected using Abbot formula (Finney, 1971) and the concentration at which 50% of the test population were dies (LC50) was determined by probit program (Finney, 1971). Results and Discussion Among the three extracts of C. nocturnum, percolation method extracts shows effective larvicidal activity over the soxhlet method (Table I). Methanol extract exhibit significant larvicidal activity causing 100% mortality in a concentration of 100 pg/mL. This extract was further fractioned and the fractions obtained (CNM1-CNM6) were tested for larvicidal activity. The fraction CNM2, 3 and 4 presented the best results (Table II). CNM1, 5, 6 fraction didn't presented good activity in concentration lower than 350 pg/mL (result not shown). The LC50 of the methanol extract and its potent fraction (CNM3) is found 14 (± 0.3113) pg/mL and 6 (± 0.1532) pg/mL respectively (LC50 values are presented as average of four observations ± SE). In all larvicidal assays, the methanol extract of C. nocturnum leaves extracted with percolation and its fractions presented higher larvicidal activity. Various authors have evaluated larvicidal activity of cestrum species on mosquitoes; where they found a steroidal bioactive compounds responsible for mosquitocidal activity (Ghosh and Chandra, 2006; Ghosh et al., 2008). Eight steroidal glycosides have been isolated from the leaves of C. nocturnum (Mimaki et al., 2002). Thus to establish relevance of these steroidal compound with the mosquitocidal activity, fractions will be further fractionated to evaluate their potential to broad use and their possible toxic effect upon the other organism. Acknowledgement Authors would like to thank Prof. M. U. Patil, Department of Zoology and Dr. Babasaheb Ambedkar, Marathwada Univer- sity, Aurangabad for providing basic training and necessary facilities to carry out the research work. References Finney DJ. Probit analysis. Cambridge (England), Cambridge University Press, 1971. Fulmali PV, Walimbe A, Mahadev PV. Spread, establishment and prevalence of dengue vector Aedes aegypti (L.) in Konkan region, Maharashtra, India. Indian J Med Res. 2008; 127: 589- 601. PMid:18765879 Ghosh A, Chandra G. Biocontrol efficacy of Cestrum diurnum L. (Solanaceae: Solanales) against the larval forms of Anopheles stephensi. Nat Prod Res. 2006; 20: 371-79. Ghosh A, Chowdhury N, Chandra G. Laboratory evaluation of phytosteroid compound of mature leaves of Day Jasmine (Solanaceae: Solanales) against larvae of Culex quinquefasciatus (Diptera:Culicidae) and nontarget organisms. Parasitol Res. 2008; 103: 271-77. Kumar K, Chhabra M, Katyal R, Patnaik PK, Kukreti H, Rai A, Saxena VK, Mittal V, Lal S. Investigation of an outbreak of Chikungunya in Malegaon municipal areas of Nashik District, Maharashtra (India) and its control. J Vector Borne Dis. 2008; 45: 157-63. Mimaki Y, Wantanabe K, Sakagami H, Sashida Y. Steroidal glycosides from the leaves of Cestrum nocturnum. J Nat Prod. 2002; 65: 1863-68. Pelah D, Abramovich Z, Markus A, Wiesman Z. The use of commercial saponin from Quillaja saponaria bark as a natural larvicidal agent against Aedes aegypti and Culex pipiens. J Ethnopharmacol. 2002; 81: 407-09. Sukumar K, Perich MJ, Boobar LR. Botanical derivatives in mosquito control: A review. J Am Mosq Control Assoc. 1991; 7: 210-37. PMid:1680152 World Heath Organization, Instructions for determining the susceptibility or resistance of mosquito larvae to insecticides. WHO/VBC. 1981; 81: 807-962. Author Info Chetan Jawale (Principal contact) e-mail: csjawale@hotmail.com 40 Bangladesh J Pharmacol 2010; 5: 39-40 Table I Lethal concentration of C. nocturnum extracts Extraction method Extracts LC100 (pg/mL) Regression equation Soxhlet Hexane a a Ethyl acetate 300 Y=2.1180X + 2.3457 Methanol 65 Y=1.9441X + 3.3592 Hexane a a Percolation Ethyl acetate 210 Y=1.7586X + 2.0573 Methanol 35 Y=1.4832X + 1.3753 aIndicate no larvicidal activity in concentration <350 µg/mL Table II Lethal concentration of fractionated methanol ex- tract of C. nocturnum Ex- tracts Hexane:Ethyl acetate LC100 (Pg/mL) Regression equa- tion CNM2 1:10 60 Y=2.0045X + 1.0231 CNM3 1:1 12 Y=1.4324X + 2.1042 CNM4 10:1 75 Y=2.2342X + 1.1103 Acknowledgement: References: DatePrinted: This article was downloaded by you on: Apr 05, 2018