The toxicity of the essential oils of eighteen Malaysian plants on the 4thinstar larvae of three vector mosquitoes (Anopheles maculatus, Aedes aegypti and Culex quinquejasciatus) was studied. The leaf oil of Litsea elliptica was the most effective, exhibiting LCso of 13.61 f-lg mr! for An. maculatus, 16.01 f-lg

ml for Ae. aegypti and 14.63 f-lg ml for C. quinquejasciatus. The essential

oils from the other species were also toxic with LCso ofl6.58 - 161.12 f-lg ml

Ahmad, Rohani

Jantan, Ibrahim

Mohd Zaki, Zaridah

Universiti Putra Malaysia Institutional Repository

Pertanika J. Sci. & Techno!. 4(1): 51-55 (1996)ISSN:0128-7680© Penerbit Universiti Pertanian MalaysiaLarvicidal Properties of the Essential Oils of SOIneMalaysian Plants on Three Vector MosquitoesIbrahim Jantan, Zaridah Mohd Zaki and Rohani Ahmad*Forest Research Institute Malaysia,Kepong, 52109 Kuala Lumpur, Malaysia*Institute for Medical Research Malaysia,50586 Kuala Lumpur, MalaysiaReceived 22 December 1994ABSTRAKCiri toksik minyak pati dari lapan belas spesies tumbuhan keatas instarkeempat larva tiga nyamuk vektor (Anopheles maculatus, Aedes aegypti danCulex quinquejasciatus) telah dikaji. Minyak pati daripada daun Litsea ellipticaadalah yang paling efektif, menunjukkan LCso 13.61 f-lg mr l keatas An.maculatus, 16.01 f-lg mr l ke atas Ae. aegypti dan 14.63 f-lg mr l ke atas C. quin-quejasciatus. Minyak-minyak pati dari species yang lain juga menunjukkannilai toksik yang tinggi, dengan LCso berkisar dari 16.58 ke 161.12 f-lg mr!.ABSTRACTThe toxicity of the essential oils of eighteen Malaysian plants on the 4th-instar larvae of three vector mosquitoes (Anopheles maculatus, Aedes aegyptiand Culex quinquejasciatus) was studied. The leaf oil of Litsea elliptica was themost effective, exhibiting LCso of 13.61 f-lg mr! for An. maculatus, 16.01 f-lgmr! for Ae. aegypti and 14.63 f-lg mr! for C. quinquejasciatus. The essentialoils from the other species were also toxic with LCso ofl6.58 - 161.12 f-lg mr!.Keywords: essential oils, Litsea elliptica, bioassay, larvicidal activities,toxicities, lllosquito larvaeINTRODUCTIONIn the search for new measures to control vector insects, the commonphenomenon of vector resistance to insecticides and the residual effects ofthese chemicals on the environment are causes for concern. The recentpublic awareness of the hazardous effects of highly toxic and non-biodegradable synthetic insecticides on human health has promptedscientists to seek safer alternatives in the form of natural products to beused directly or as starting materials to synthesize more potent derivativesor as models for the developement of synthetic chemicals.Ibrahim Jantan, Zaridah Mohd Zaki and Rohani AhmadNatural products of plant origin such as rotenone, nicotine andpyrethrins have long been used to control destructive insects and vectors ofdiseases (Matsumura 1975). One of the earliest reports on the toxicity ofplant extracts on mosquito larvae was by Campbell and Sullivan (1933)who reported that the plant alkaloids, nicotine, anabasine, methylanabasineand lupinine killed larvae of Culex pipiens, Cx. territans and Cx.quinquefasciatus. Subsequently, many researchers have reported on theeffectiveness of plant extracts against mosquito larvae (Haller 1940; Hartzelland Wilcoxon 1941; Amonkar and Reeves 1970; Supavan et al. 1974;Chavan 1983). Recently, Zebitz (1986) reported that neem seed kernelextract (Azadirachta indica) is active against 4th-instar larvae of Aedes togoiand Ae. aegypti with LCso of 1.19 - 18.10 f.tg mr l .This paper reports on the toxic effects of the extracts of eighteenMalaysian plants on 4th-instar larvae of Anopheles maculatus, Aedes aegypti andCulex quinquefasciatus.MATERIALS AND METHODSPlant materials were collected from various locations in PeninsularMalaysia. The plants were identified and voucher specimens weredeposited at the herbarium of Forest Research Institute Malaysia. Theessential oils were prepared by subjecting the plant materials (200 g, meshsize 40 - 60) to water distillation for 8 h.Three species of vector mosquitoes, Anopheles maculatus, Aedes aegypti andCulex quinquefasciatus, served as the test organisms. The larvae colonies ofthese mosquitoes were established and collected from the Insectary of theInstitute for Medical Research, Kuala Lumpur.Each essential oil in 0.2 ml ethanol was dissolved in distilled water toprepare 1000 f.tg ml-l stock solution from which concentrations of 500, 200,150, 100, 50 and 1°f.tg mr l were prepared by dilution. Twenty ml of eachsample were placed in a vial. Ten 4th-instar larvae of each vector specieswere transferred into each vial using a disposable pipette. The bioassay wascarried out in two stages. Initially, all plant extracts were screened usingconcentrations of 1000 and 500 f.tg mr l . Extracts producing high mortalityrates were further tested at lower concentrations of 200, 150, 100,50 and 10f.tg mr l . The treatment on each concentration was replicated 3 times in acompletely randomized design. A control sample was prepared by theaddition of 0.2 ml ethanol to the water in each vial which contained tenlarvae. Solutions of DDT dissolved in water at 1 - 200 f.tg mr lconcentrations were used as a standard toxicant.Mortality was observed after 24 h and corrected mortality obtained byapplying Abbott's formulae (1925). LCso and 95% confidence intervalswere determined by the probit analysis method as described by Finney(1971).52 Pertanika J. Sci. & Techno!. Vo!. 4 No. I, 1996Larvicidal Properties of the Essential Oils of Some Malaysian Plants on Three Vector MosquitoesRESULTS AND DISCUSSIONThe bioassay of the essential oils against mosquito larvae demonstrated theireffectiveness, with LCso of 13.6 - 161.1 J.Lg mr l (Table 1). However, theefficacy of each oil towards the various larvae was non-selective as LCsoshowed little variation. The only exception was the essential oil ofGoniothalamus andersonii which showed relatively strong activity againstCulex quinquifasciatus (LCso 60.85 J.Lg mr l ), but weaker activity against theother two vectors with LCso of 113.85 and 116.85 J.Lg mr! respectively.TABLE 1Toxicity of essential oils of some Malaysian plants to mosquito larvaeSample LC50 (95% CI)A.e. C.q. A.m.Cinnamomum 116.85 121.32impressicostatum (111.09 - 122.40) (110.79 - 136.03)Cinnamomum 119.25 132.85mollisimum (110.10 - 129.38) (125.47 - 140.64)Cinnamomum 62.91 63.07mers (59.42 - 66.80) (57.19 - 68.50)Cinnamomum 87.47 72.71zeylanicum (77.94 - 97.79) (67.02 - 79.24)Piper aduncum 23.38 16.58 18.97(21.07 - 25.76) (15.69 - 17.38) (16.94 - 20.96)Piper betle 75.67 59.01 74.00(67.36 - 82.88) (53.93 - 62.82) (69.59 - 79.44)Piper penangense 161.12 128.83 102.59(150.62 - 169.82) (122.05 - 135.09) (84.82 - 122.49)Piper lanatum 82.18 92.32 62.14(78.91 - 85.30) (86.99 - 100.43) (57.83 - 66.65)Piper pedicellosum 84.44 70.25 49.03(77.45 - 93.97) (67.82 - 72.85) (31.63 - 62.06)Litsea elliptica 16.01 14.63 13.61(14.29 - 18.05) (13.85 - 15.79) (10.79 - 15.59)Goniothalamus 116.85 60.93 113.85andersonii (111.09 - 122.40) (50.81 - 69.73) (98.12 - 126.16)Pertanika J. Sci. & Techno!. Vo!. 4 No. I, 1996 53Ibrahim Jantan, Zaridah Mohd Zaki and Rohani AhmadTable 2: Cont'dLeptospermum 68.51 68.38 91.15javanicum (63.75 - 73.41) (64.43 - 71.63) (75.57 - 119.53)Pogostemon cablin 67.46 56.01(61.85 - 72.72) (50.36 - 61.07)Cymbopogon nardus 74.51 71.09(67.23 - 83.03) (67.41 - 74.64)Melaleuca cajupati 82.04 60.29(79.83 - 84.12) (57.61 - 62.81)Vetiveria zizanioides 90.27 95.85(77.30 - 105.39) (88.19 - 103.22)Polygonum minus 47.94 38.43(43.23 - 52.79) (33.75 - 44.05)Dipterocarpus kerrii 146.10 126.83(139.66 - 152.29) (110.57 - 136.38)*Note:A.e. = Aedes aegyptic.q. = Culex quinquefasciatusA.m. = Anopheles maculatusThe essential oils of Litsea elliptica, Piper aduncum and Polygonum minusshowed LCso < 50 f-Lg mr!, indicating significant levels of larvicidalproperties. The leaf oil of Litsea elliptica plant extract was the most effective,exhibiting LCso of 13.61 f-Lg mr l for Anopheles maculatus, 16.01 f-Lg mr! forAedes aegypti and 14.63 f-Lg mr l for Culex quinquifasciatus; this showed that theplant extracts contained active principles responsible for the larvicidalactivity.The active principles of the essential oils, when isolated in pure form,might possess high larvicidal activity. The results should encourage furtherefforts to purify the active constituents and study their pathological effectson mosquito larvae. The toxic properties of the essential oils could also bedue to the combined effect of the compounds in the crude extracts whichwere of diverse chemical structures and could exhibit a different mode ofaction towards the test organisms, resulting in high toxicities.CONCLUSIONThe results indicate that some of the plants studied can be effectively used inmosquito control as an alternative to synthetic insecticides. Although theplant extracts were less toxic than the chlorinated insecticide, DDT, they54 Pertanika J. Sci. & Techno!. Vol. 4 No.1, 1996Larvicidal Properties of the Essential Oils of Some Malaysian Plants on Three Vector Mosquitoesare environment-friendly and less harmful than the latter, which has beenreported to cause undesirable side effects to human (Reynolds 1989).However, the residual lifespan as well as their performance under fieldconditions need to be determined to assess their potential as commercialinsecticidal agents. The leaf oil of Litsea elliptica with the lowest value ofLCso has the greatest potential, followed by the leaf oils of Polygonum minusand Piper aduncum which showed LCso < 50 p,g mr l . The active ingredientsof each extract and the minimum amount needed in formulations need to bedetermined.ACKNOWLEDGEMENTThe authors are grateful to the Director of Institute of Medical Research,Malaysia, for providing the facilities and Abdul Rashih Ahmad and AbuSaid Ahmad of the Forest Research Institute Malaysia for their technicalassistance.REFERENCESABBOTT, W.S. 1925. A method of counting the effectiveness of an insecticide. J. Econ.Entomol. 18: 265-267.AMONKAR, S.V. and E.C. REEVES. 1970. Mosquito control with active principle ofgarlic, Allium sativum. Journal of Economic Entomology 63: 1172-1175.CAMPBELL, F.C. and W.N. SULLIVAN. 1933. The relative toxicity of nicotine,anabasine, methyl anabasine and lupinene for culicine mosquito larvae. Journal ofEconomic Entomology 26: 500-509.CHAVAN, S.R. 1983. Chemistry of alkanes separated from leaves of Az:.adirachta indicaand their larvicidal/insecticidal activity against mosq uitoes. In Proceedings 2nd.International Neem Conference, Rauischholzhausen. p. 59-65.FINNEY, D.J. 1971. Probit AnalYsis: A Statistical Treatment of the Sigmoid Response Curve. 3rdedn. Cambridge University Press.HALLER, H.L. 1940. Insecticidal properties of the fruit of Phellodendron spp. J. Econ.Entomol. 33: 941.HARTZELL, A. and F. WILCOXON. 1941. A survey of plant products for insecticidalproperties. Contrib. Boyce Thompson Inst. 12: 127-141.MATSUMURA, F. 1975. Toxicology of Insecticides. Tew York: Plenum Press.REYNOLDS, ].F. 1989. Martindale, The Extra Pharmacopoeia. London: PharmaceuticalPress.SUPAVARN, P., F.W. KNAPP, and R. SIGATUS. 1974. Biologically active plant extractsfor control of mosquito larvae. Mosquito News 34: 398-402.ZEBITZ, C.P.W. 1986. Potential ofneem seed kernel extracts in mosquito control. InProceedings 3rd. International Neem Coriference, Nairobi. p. 555-573.Pertanika J. Sci. & Technol. Vol. 4 No. I, 1996 55

Larvicidal Properties of the Essential Oils of Some

Malaysian Plants on Three Vector Mosquitoes

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Larvicidal Properties of the Essential Oils of Some Malaysian Plants on Three Vector Mosquitoes

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