Larvicidal efficacy of Ficus benghalensis L. plant leaf extracts against Culex quinquefasciatus Say, Aedes aegypti, Anopheles stephensi.

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1 Europen Review for Medicl nd Phrmcologicl Sciences 2010; 14: Lrvicidl efficcy of Ficus benghlensis L. plnt lef extrcts ginst Culex quinquefscitus Sy, Aedes egypti L. nd Anopheles stephensi L. (Dipter: Culicide) M. GOVINDARAJAN Division of Vector Biology nd Phytochemistry, Deprtment of Zoology, Annmli University, Annmlingr (Indi) Abstrct. Objective: To investigte the lrvicidl efficcy of different extrcts of Ficus benghlensis L. ginst Culex quinquefscitus Sy, Aedes egypti L. nd Anopheles stephensi L. ( Dipter: Culicide). Mterils nd Methods: Lrvicidl efficcy of the crude lef extrcts of Ficus benghlensis with three different solvents like methnol, benzene nd cetone ws tested ginst the erly second, third, fourth instr lrve of Culex quinquefscitus, Aedes egypti nd Anopheles stephensi. Results: The lrvl mortlity ws observed fter 24 h of tretment. Among the three solvents the mximum efficcy ws observed in methnol. The letl concentrtion (LC 50 ) vlues of Ficus benghlensis ginst erly second, third nd fourth lrve of Culex quinquefscitus, Aedes Aegypti nd Anopheles stephensi were 41.43, nd ppm, 56.54, nd ppm nd 60.44, nd ppm respectively. No mortlity ws observed in controls. The chi-squre vlue were significnt t p <0.05 level. Conclusion: The methnol extrct of F. benghlensis showed good lrvicidl ctivity ginst three vector mosquitoes. Key Words: Ficus benghlensis, Lrvicidl ctivity, Culex quinquefscitus, Aedes egypti, Anopheles stephensi. Introduction Mosquitoes re not only the most importnt vectors for the trnsmission of mlri, filrisis, nd virl diseses 1, but re lso n importnt pest to humns, cusing llergic responses tht include locl skin rection nd systemic rections such s ngioedem, nd urticri 2. Control of the mosquito lrve is frequently dependent on continued pplictions of orgnophosphtes (chlorpyrifos, temephos, nd fenthion) nd insect growth regultors (diflubenzuron nd methoprene) 3. Effective, repeted use of controlling gents hs disrupted nturl biologicl control systems nd led to outbreks of insect species showing pesticide resistnce. It hs lso provoked undesirble effects, including toxicity to non-trget orgnisms nd fostered environmentl nd humn helth concerns 4. These problems hve highlighted the need for the development of new strtegies for selective mosquito lrvl control. Plnts my be source of lterntive gents for control of mosquitos, becuse they re rich in bioctive chemicls, re ctive ginst limited number of species including specific trget insects, nd re bio-degrdble. They re potentilly suitble for use in integrted pest mngement progrms 5. The plnt product s phytochemicl is used s insecticide for killing lrve or dult mosquitoes or s repellent for protection ginst mosquito bites. There re more thn 800 species nd 2000 vrieties of Ficus benghlensis L. species, (Morcee, Mulberry fmily) most of which re ntive to the old World tropics. Ficus benghlensis remrkble tree of Indi sends down its brnches nd gret number of shoots, which tke root nd become new trunk. This tree is considered to be scred in mny plces in Indi. Erlier glucoside, 20-tetrtriconthene-2-one, 6-hepttriconthene- 10-one, penttricontn-5-one, bet sitostirollph-d-glucose nd meso-inositol hve been iso- Corresponding Author: Mrimuthu Govindrjn, M.Sc., M.Phil., Ph.D.: e-mil: drgovind1979@rediffmil.com 107

2 M. Govindrjn lted from the brk of Ficus benghlensis 6 Leves contin crude protein 9.63%, crude fibres-26.84%, CO-2.53%, nd phosphorus-0.4%. It yields ltex contining coytchoue (2.4%), resin, lbumin, cerin, sugr nd mlic cid. It hs been used for the tretment of dirrhoe, dysentery nd piles 7, rheumtism, skin disorders like sores 8, to boost immune system 9, s hypoglycemic 10. The extrcts of Ficus benghlensis were lso reported to inhibit insulinse ctivity from liver nd kidney 11. The im of the present study ws to evlute the lrvicidl ctivity of different extrcts of Ficus benghlensis ginst Culex quinquefscitus, Aedes egypti nd Anopheles stephensi. Mterils nd Methods Plnt Collection The Ficus benghlensis fresh leves were collected from in nd round Vittloor, Thnjvur district, Tmilndu, Indi. It ws uthenticted by plnt txonomist from the Deprtment of Botny, Annmli University. A voucher specimen is deposited t the herbrium of plnt phytochemistry division, Deprtment of Zoology, Annmli University, Indi. Preprtion of the Extrct The fresh leves were wshed with tp wter, shde dried t room temperture (28 ± 2 C) nd powdered by electricl blender (Preethi, Soln, Indi). The powder (1.0 kg) ws extrcted with three solvents, such s methnol, benzene nd cetone (3 L) t Soxhlet pprtus (Hitech, Puducherry, Indi) for 8 h followed by the stndrd procedure 12. The extrcts were filtered through Buchner funnel (Borosil, Mumbi, Indi) with Whtmn number 1 filter pper. The filtrte ws evported to dryness under reduced pressure using rotry evportor (Superfit, Indi/Model-PBU-6D, Mumbi, Indi) to yield drk greenish, gummy extrct. The yield of crude extrct ws 26.1, 28.0 nd 25.4 gms in methnol, benzene nd cetone, respectively. Stndrd stock solutions were prepred t 1% by dissolving the residues in cetone, which ws used for the biossys. Test Orgnisms The mosquitoes, Culex quinquefscitus, Aedes egypti nd Anopheles stephensi were rered in the lbortory. The lrve were fed on dog biscuits nd yest power in the 3:1 rtio. Adults were provided with 10% sucrose solution nd one week old chick for blood mel. Mosquitoes were held t 28 ± 2, 70-85% reltive humidity (RH), with photo period of 14 h light, 10 h drk. Lrvicidl Biossy The lrvicidl ctivity of the lef extrct of Ficus benghlensis ws evluted s per the method recommended by WHO 13. Different concentrtion of the test smples were used. 500 ml glss beker contining 250 ml of tp wter. Erly second, third nd fourth instr of Culex quinquefscitus, Aedes egypti nd Anopheles stephensi 25 were introduced to ech of the test solution s well s control. Control consists of cetone only. For ech experiment six replictes were mintined t time. The letl concentrtion (LC 50 ) vlues were clculted fter 24 h by probit nlysis 14. Sttisticl Anlysis Sttisticl evlution ws done using Sttisticl Pckge of Socil Sciences (SPSS) 13.0 for windows, Significnce level ws set t p <0.05. Results The toxicity of Ficus benghlensis ws tested ginst the erly second, third nd fourth lrve of Culex quinquefscitus, Aedes egypti nd Anopheles stephensi. The dt were recorded nd ststicl dt regrding the LC 50, LC 90, regression eqution, Chi-squre nd 95% confidence limits were clculted (Tbles I, II, III.). The methnolic extrct of Ficus benghlensis showed highest lrvicidl ctivity ginst Culex quinquefscitus, Aedes egypti nd Anopheles stephensi. The LC 50 vlues of Ficus benghlensis ginst erly second, third nd fourth lrve of Culex quinquefscitus, Aedes egypti nd Anopheles stephensi were 41.43, nd ppm, 56.54, nd ppm nd 60.44, nd ppm respectively. No mortlity ws observed in control. The chisqure vlue were significnt t p <0.05 level. In the present study second instr ws highly sensitive when compred with third nd fourth instr of the three vector mosquitoes. 108

3 Ficus benghlensis L. plnt lef extrcts ginst Culex quinquefscitus Sy, Aedes egypti L. Tble I. Lrvicidl ctivity of Ficus benghlensis ginst different lrvl stges of Culex quinquefscitus. 2 nd Instr Methnol Benzene Acetone rd Instr Methnol Benzene Acetone th Instr Methnol Benzene Acetone Significnt t P<0.05 level. LC 50 : letl concentrtion; LCL: lower control limit; UCL: upper control limit. Discussion The present findings corporte erlier findings of Mcedo et l. 15 who showed tht ethnol extrct of Tgetes ptul ws less ctive nd only 50% lrve were killed t higher concentrtion (100 ppm). Methnolic lef extrct of Cssi fistul ws tested for lrvicidl ctivity ginst Culex quinquefscitus nd Anopheles stephensi with LC 50 vlues of nd mg/l, respectively 16. The petroleum ether frction of Acci nolotic nd Citrullus colocynthis showed 100 per cent mortlity in 100, 250 nd 500 ppm nd 60 nd 50 per cent mortlity t 125 nd 62.5 ppm respectively ginst Culex quinquefscitus 17. The lef extrct of Aclyph in- dic with different solvents viz, benzene, chloroform, ethyl cette nd methnol were tested for lrvicidl, ovicidl ctivity nd oviposition ttrctncy ginst Anopheles stephensi. The lrvl mortlity ws observed fter 24 h exposure. The LC 50 vlues re 19.25, 27.76, nd ppm, respectively 18. The lef extrct of Cssi fistul with different solvents viz, methnol, benzene nd cetone were studied for the lrvicidl, ovicidl nd repellent ctivity ginst Aedes egypti. The 24 h LC 50 concentrtion of the extrct ginst Aedes egypti were observed t 10.69, nd mg/l respectively 19. Cheng et l. 20 reported tht the lef nd brk essentil oil of Cryptomeri jponic showed lrvicidl ctivity ginst Aedes egypti. Tble II. Lrvicidl ctivity of Ficus benghlensis ginst different lrvl stges of Aedes egypti. 2 nd Instr Methnol Benzene Acetone rd Instr Methnol Benzene Acetone th Instr Methnol Benzene Acetone Significnt t P<0.05 level. LC 50 : letl concentrtion; LCL: lower control limit; UCL: upper control limit. 109

4 M. Govindrjn Tble III. Lrvicidl ctivity of Ficus benghlensis ginst different lrvl stges of Anopheles stephensi. 2 nd Instr Methnol Benzene Acetone rd Instr Methnol Benzene Acetone th Instr Methnol Benzene Acetone Significnt t P<0.05 level. LC 50 : letl concentrtion; LCL: lower control limit; UCL: upper control limit. Methnolic frction of leves of Ment piperit, Phyllnthus niruri nd Letiot sper exhibited the LC 50 vlues of 43.65, nd respectively ginst the lrve of Culex quinquefscitus 21. Singh et l. 22 reported the mosquito lrvicidl properties of the lef extrct of herbceous plnt Ocinum cnum ginst Aedes egypti. The LC 50 vlues for 2 nd, 3 rd nd 4 th lrve were , nd ppm respectively. Gusmo et l. 23 reported tht the extrct of Derris urucu (Lonchocrpus) showed lrvicidl ctivity ginst Aedes egypti with LC 50 vlues of 17.6 µg/ml. Muthukrishnn et l. 24 reported tht ethyl cette frctions of Solnum trilobtum nd Letiot sper showed the LC 50 vlues of 23.5 nd ppm ginst 2 nd nd 3 rd lrve of Culex quinquefscitus. Yng et l. 3 reported tht Piper longum fruit-isolted pipernonline hd strong lrvicidl effects ginst the 4 th stge lrve of Aedes egypti. The LC 50 vlue of pipernonline ws 0.25 mg/l ginst Aedes egypti. Kbru nd Gichi 25 reported tht the mngrove plnt Rhizophor mucront brk nd pith extrct showed high toxicity with LC 50 vlues of nd ppm respectively ginst Aedes egypti lrve. In conclusion, the present study showed tht extrcts from Ficus benghlensis cn be effectively used in the control of Culex quinquefscitus, Aedes egypti nd Anopheles stephensi mosquito lrve. However, further studies on the identifiction of the ctive principles involved nd their mode of ction nd field trils need presently other investigtions. References 1) JAMES AA. Mosquito moleculr genetics: the hnds tht feed bite bck. Science 1992; 257: ) PENG Z, YANG J, WANG H, SIMONS FER. Production nd chrcteriztion of monoclonl ntibodies to two new mosquito Aedes egypti slivry proteins. Insect Biochem Mol Biol 1999; 29: ) YANG YC, LEE SG, LEE HK, KIM MK, LEE SH, LEE HS. A piperidine mide extrcted from Piper longum L. fruit shows ctivity ginst Aedes egypti mosquito lrve. J Agr Food Chem 2002; 50: ) LEE SE, KIM JE, LEE HS. Insecticide resistnce in incresing interest. Agr Chem Biotechnol 2001; 44: ) ALKOFAHI A, RUPPRECHT JK, ANDERSON JE, MCLAUGHLIN JL, MIKOLAJCZAK KL, SCOTT BA. Serch for new pesticides from higher plnts. In: Arnson JT, Philogene BJR, Mornd P (Eds.), Insecticides of Plnt Origin. In: ACS Sym. Ser., 387. Am Chem Soc, Wshington, DC, 1989, pp ) SUBRAMANIAN PM, MISRA GS. Chemicl constituents of Ficus benghlensis. Pol J Phrmcol Phrm 1978; 30: ) MUKHERJEE PK, SAHA K, MURUGESAN T, MANDAL SC, PAL M, SAHA BP. Screening of ntidirrhoel profile of some plnt extrcts of specific region of West Bengl. Indi J Ethnophrm 1998; 60: ) WARRIER PK, NAMBIAR VPK, RAMANKUTTY C. Indin Medicinl Plnts, (1-5) Orient Longmn Ltd., Mdrs, ) GABHE SY, TATKE PA, KHAN TA. Evlution of the immunomodultory ctivity of the methnol extrct of Ficus benghlensis roots in rts. Indin J Phrmcol 2006; 38:

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