Research ArticleInsecticidal Activity and Chemical Compositionof the Morinda lucida Essential Oil againstPulse Beetle Callosobruchus maculatus
Moses S Owolabi1 Eduardo Padilla-Camberos2 Akintayo L Ogundajo1
Isiaka A Ogunwande1 Guido Flamini3 Olaniyi K Yusuff1 Kirk Allen2
Karen Isabel Flores-Fernandez4 and Jose Miguel Flores-Fernandez2
1 Department of Chemistry Lagos State University PMB 0001 Lasu Post Office Ojo Lagos 102001 Nigeria2 Biotecnologıa Medica y Farmaceutica Centro de Investigacion y Asistencia en Tecnologıa y Diseno del Estado de JaliscoAC Avenida Normalistas 800 Col Colinas de la Normal 44270 Guadalajara JAL Mexico
3 Dipartimento di Chimica Bioorganica e Biofarmacia Universita di Pisa Via Bonanno 33 56126 Pisa Italy4 Ingenierıa en Biotecnologıa Universidad Politecnica del Valle de Toluca Carretera Toluca-Almoloya de Juarez km 56Camino la Loma Santiaguito Tlalcilalcali 50900 Almoloya de Juarez Mexico
Correspondence should be addressed to Moses S Owolabi sunnyconcept2007yahoocomand Jose Miguel Flores-Fernandez jmffiiahotmailcom
Received 22 January 2014 Accepted 4 July 2014 Published 20 July 2014
Academic Editor Alexsandro Branco
Copyright copy 2014 Moses S Owolabi et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited
Insecticidal activity of essential oil extracted from Morinda lucida was tested on pulse beetle Callosobruchus maculatus which isa pest that causes serious damage to several pulses The insecticidal activity was compared with two pesticides Phostoxin andPrimo-ban-20 120 mixed sex adult C maculatus were introduced along with 30 g of cowpeas Four concentrations (040 020010 and 005120583gmL) of theM lucida essential oil Phostoxin and Primo-ban-20 were tested Essential oil chemical compositionwas analyzed by GC-MS M lucida essential oil showed a high toxicological effect producing 100 mortality after 72 hours at adose of 020 120583gmL M lucida essential oil had a potent insecticidal activity (LC
90= 0629 120583gmL) compared to both pesticides
Phostoxin (LC90= 0652 120583gmL) and Primo-ban-20 (LC
90= 0726 120583gmL) at 24 h The main compounds of the essential oil were
the oxygenated monoterpenoids 18-cineole (434) and 120572-terpinyl acetate (145) and the monoterpene hydrocarbons mostlysabinene (82) and 120573-pinene (40) Results clearly indicate thatM lucida essential oil can be used as an effective alternative forpulse beetle C maculatus control and it could be tested against other pulse beetles affecting Asia and Africa and throughout theworld thereby reducing use of synthetic pesticides
1 Introduction
Callosobruchus maculatus is an economically important pestof several pulses including leguminous grains such as lentilscowpeas green gram chickpea black gram soybean andharicot beans [1 2] These pulses are important sourcesof vegetable protein for millions of people of tropical andsubtropical regions of Asia and Africa C maculatus larvaebore into pulse grain or into stored crop making it unfit forhuman consumption and unviable for replanting [3ndash5]
Control of insect pestsrelies heavily on the use of syntheticinsecticides such as methyl bromide or phosphine Howevertheir intensive use has led to the development of geneticresistance by insect species pest resurgence residual toxicityenvironmental hazards and serious problems arising fromfactors such as direct toxicity to predators pollinators fishand man [6] As such the use of methyl bromide is beingrestricted because of its potential to damage the ozone layer[7] Susceptibility of crop plants to insect pests and increasingcosts of application of the presently used synthetic pesticides
Hindawi Publishing Corporatione Scientific World JournalVolume 2014 Article ID 784613 7 pageshttpdxdoiorg1011552014784613
2 The Scientific World Journal
have directed the need for effective biodegradable pesticides[8 9]
Because of their high volatility plant extracts or plantcompounds and the use of natural compounds with insec-ticidal activity provide a potential biodegradable alternativeto synthetic pesticides [10ndash12] The contact and fumigantinsecticidal activity of several plant essential oils and theirconstituents have been demonstrated against stored-productpests [13ndash15] a number of bruchid pests [16ndash18] and insectsand mites [19]
Several botanical families for pest control are describedsuch as Meliaceae Rutaceae Asteraceae Labiatae and Mal-vaceae Rubiaceae is a family of 630 genera and about13000 species [20] Morinda lucida (Benth) a member ofRubiaceaefamily is a medicinal plant of about 15m tall with adense crown of slender crooked branches approximately 20ndash30 cm in diameterM lucida commonly known by the Yorubain southwestern part of Nigeria as ldquoOruwordquo is widely dis-tributed in West Africa and is used in African folk medicineto treat several diseases [21] The leaves are bitter and areused by the natives to treat malaria yellow fever jaundicehepatitis eczema edema cough hypertension diabetes andsexual weakness [20 22 23]
In a previous study another species of the same genusM citrifolia showed larvicidal activity against three species ofmosquito vectors [24] and the biofriendly nature ofM lucidasuggested the need to search for new properties of this planttherefore the present study was conducted to determine theinsecticidal activity of the essential oil of M lucida againstC maculatus by dose response bioassays and to characterizeits chemical composition for consideration as an alternativeinsecticide for pulse beetle
2 Materials and Methods
21 Plant Material The aerial parts of the M lucida werecollected fromApomu (7∘19101584048N 4∘10101584047E) which is locatedin Osun State Nigeria and identified at the Herbarium ofthe Botany Department University of Lagos Nigeria wherevoucher specimen number LUH 2637 was kept The sampleswere air-dried pulverized and stored in polythene bag toreduce evaporation of the volatile oil
To extract the essential oil 500 g of pulverized sample wasplaced in a 5 L flask and distilled water was added to coversample Essential oil was obtained by hydrodistillation usingglass modified Clevenger-type apparatus the process ran for4 h at normal atmospheric pressure and at 96-97∘C inside theextractor After the essential oil isolation residual water wasremoved by filtration with anhydrous sodium sulphate andthen the essential oil was stored in an amber vial at 4∘C forfuture analysis
22 Gas Chromatography (GC) Analysis GC analysis wasperformed with a HP-5890 Series II instrument equippedwith a HP-Wax and HP-5 capillary columns (both 30m times025mm ID times 025 120583m film thickness) working with thefollowing temperature program 60∘C for 10min rising at5∘Cmin to 220∘C The injector and detector temperatures
were maintained at 250∘C carrier gas nitrogen (2mLmin)detector dual FID split ratio 1 30 The volume injectedwas 05120583L The relative proportions of the oil constituentswere percentages obtained by FID peak-area normalizationwithout the use of response factor
23 Gas Chromatography-Mass Spectrometry (GC-MS) GC-MS analysis was performed with a Varian CP-3800 gaschromatograph equippedwith aHP-5 capillary column (30mtimes 025mm ID times 025 120583m film thickness) and a Varian Saturn2000 ion trap mass detector Analytical conditions wereinjector and transfer line temperature 220∘C and 240∘Crespectively oven temperature was programmed from 60 to240∘C at 3∘Cmin Carrier gas was helium at 10mLmin flowrate Injection volumewas 02120583L (10hexane solution) usinga 1 30 split ratio The mass spectrometer was operated at 70-eV ionization voltage The acquisition mass range was 30 to300119898119911 at 10 scans
24 Identification of the Volatile Compounds The identifica-tion of the volatile constituents of the essential oil sample wasmade using the following criteria (1) comparison of massspectra with the Wiley library spectra 275 L (RevC0000)electronic database (2) injection of authentic compoundsfrom Sigma-Aldrich with a 95 minimum purity under thesame analytical conditions and (3) comparison of the reten-tion index (RI) obtained by GC-MS with the RI theoreticalin similar phases The molecular weights of all the identifiedsubstances were confirmed by GC-MS using MeOH as CIionizing gas
25 Insect Culture Callosobruchus maculatus were reared oncowpea seeds (Vigna unguiculata) under controlled temper-ature and humidity at 28 plusmn 1∘C 58 plusmn 5 RH (12 h light cycle)and 255 plusmn 1∘C 45plusmn 5RH (12 h dark cycle) Adult insects of8 days old were used for insecticidal test
All experimentswere carried out under the same environ-mental conditions as described above
26 Insecticidal Activity of M lucida Essential Oil The insec-ticidal activity was evaluated as described by Ilboudo et al[25] One hundred and twenty mixed sex adult C maculatus(8 days old) were put into a 500mL glass bottle with 30 g ofcowpea seeds and kept in the laboratory at 28 plusmn 1∘C 58 plusmn 5RH (12 h light cycle) and 255 plusmn 1∘C 45 plusmn 5 RH (12 h darkcycle) for 72 h Concentrations of the M lucida essential oildiluted in acetone were tested on C maculatus (005 010020 and 040 120583gmL) The appropriate concentrations wereapplied to filter paper (Whatman number 1 cut into 7 cmdiameter) and immediately introduced into a glass bottlethat was then hermetically sealed For the control groupthe insects were placed in the glass bottles under the sameconditions but without the essential oil Each concentrationand control was replicated three times Insect mortalitywas determined by observing the recovery of immobilizedinsects in 12 h intervals up to 72 h When no antennal or legmovements were observed insects were considered dead
The Scientific World Journal 3
27 Insecticidal Activity of Commercial Pesticides Two syn-thetic pesticides were purchased from Chemical and Alliedcompany Lagos Nigeria Phostoxin (55 Aluminium phos-phide) and Primo-ban-20 (pirimiphos-methyl emulsifiableconcentrate 50) The first is converted to phosphine gaswhich is taken by the insect through its respiratory systemand the second is able to penetrate the insect cuticle Pelletsof Phostoxin were divided into small pieces and weighedto achieve the appropriate concentrations and were appliedaccording to manufacturerrsquos directions [26] Primo-ban-20was diluted to achieve the desired concentrations from astock solution of 2 120583gmL Both pesticides were evaluated atthe same concentrations as the essential oil in triplicate
28 Mortality Determination Ten pairs of adultCmaculatuswere released each 12 h after exposure of essential oil upto 72 h After a 12 h exposure period the containers wereopened and the dead beetles were removed and recordedKnocked-down adults were regarded as alive if they showedcontinued movement of their appendages by the touch ofa fine brush After observation the containers were tightlyclosed After every 12 h new batches of twenty insects werereleased and the numbers of dead insects were recorded andthe percentage of mortality was calculated and correctedusing Abbottrsquos formula [27]
29 Data Analysis Average percentage insect mortality wascalculated from three replicates Dosage-mortality responseswere used to estimate LC
50(concentration causing 50
mortality) and LC90
(concentration causing 90 mortality)values at 24 and 48 h with Probit analysis [28] using Stat-graphics 51 software Insecticidal activity was consideredsignificantly different when the 95 CI (confidence interval)fail to overlap
3 Results
31 Chemical Composition of Morinda lucida The chemicalcomposition analysis by GC-MS identified 63 components ofM lucida essential oil containing mainly 18-cineole (434)followed by 120572-terpinyl acetate (145) sabinene (82) and120573-pinene (40 Table 1)
32 Mortality of Callosobruchus maculatus The aerial partsof the M lucida distillate yielded a 048 (vw) essentialoil The insecticidal bioassay of the essential oil of M lucidaagainstC maculatus (Figure 1) showed that the activity of theoil was both dose dependent and exposure dependent At adose of 005120583gmL the essential oil produced 594 mortal-ity after 72 h (Figure 1(c)) The essential oil produced 333567 and 899 mortality after 24 48 and 72 h at a dose of010 120583gmL respectively while a dose of 020120583gmL yieldedmortality of 467 833 and 100 respectively over thesame time duration (Figure 1) The highest concentration of040 120583gmL produced a mortality of 90 and 100 after 48and 72 h respectively (Figures 1(b) and 1(c))
The synthetic pesticide Primo-ban-20 at 020120583gmL hada lower mortality (367) at 24 h while Phostoxin had a
Table 1 Retention index and relative composition of major chemi-cal components ofM lucida essential oil
Component KIa 120573-Caryophyllene 1421 Tr119901-Cymen-7-ol-acetate 1423 Tr(119864)-Cinnamyl acetate 1450 Tr(119864)-Methyl isoeugenol 1495 TrBicyclogermacrene 1497 Tr120572-Bulnesene 1506 Tr120573-Sesquiphellandrene 1526 TrElemol 1554 TrElemicin 1557 TrSpathulenol 1578 04Caryophyllene oxide 1583 05g-Eudesmol 1635 Tr120573-Eudesmol 1651 02120572-Eudesmol 1655 TrIntermedeol 1667 TrTotal identified 9540Major components are shown in boldlowastThese chemical components were identified by authentic compound injec-tionaKI Kovats retention indicesTr = trace amount lt01
467 and 867mortality after 24 h and 48 h at 020 120583gmLrespectively (Figures 1(a) and 1(b)) Both M lucida essen-tial oil and pesticides reached 100 mortality at 72 h at020120583gmL (Figure 1(c)) The negative control showed noactivity except at 72 h (17)
33 Determination of Insecticidal Activity Probit analysisshowed that M lucida essential oil after 24 h presentedan insecticidal activity (LC
90= 0629120583gmL) similar to
Phostoxin (LC90
= 0652120583gmL) and Primo-ban-20 (LC90
= 0726 120583gmL) At 24 h essential oil of M lucida (LC50
= 0298 120583gmL) and Phostoxin (LC50
= 0295 120583gmL) weremore efficient than Primo-ban-20 (LC
50= 0393 120583gmL) At
48 h after exposure there was no significant difference inLC50
between M lucida essential oil compared to Primo-ban-20 and Phostoxin (Table 2 where all confidence intervalsoverlap)
4 Discussion
Recently biological activity has been demonstrated for theM lucida essential oil such as the complete suppression ofaflatoxin synthesis in maize [29] cytotoxic activity on cancercell lines [30] and antioxidant and anti-inflammatory activity[20] This study demonstrated a potent insecticidal activityfor M lucida showing 100 mortality on C maculatus atconcentrations of 020 and 040120583gmL after 72 h (Figure 1)
Our results for M lucida essential oil are consistent withother reports of essential oils that exhibited insecticidal andrepellency activity against C maculatus such as Eucalyp-tus intertexta Callistemon viminalis Ocimum americanum
Hyptis suaveolens Hyptis spicigera Lippia multiflora Cym-bopogon citratus and Lippia rugosa [25 31ndash35] Chaubey[36] evaluated the insecticidal oviposition egg hatchingand developmental inhibitory activities of seven differentessential oils against Callosobruchus chinensis and found thatNigella sativa was the most effective at all stages MoreoverKeita et al [16] found that essential oils ofOcimum basilicumand O gratissimum at a dose of 25 120583gmL after 12 h offumigation produced 80 and 70 mortality respectivelySimilarly Moharramipour et al [37] have shown a repellencyof 824 of the Thymus persicus essential oil against Cmaculatus
The chemical components of this essential oil have beenpreviously reported [38 39] although there were differencesin the ratio of their chemicals constituents Okoh et al [38]identified 50 compounds where 120572-terpinene was the majorconstituent with 178 while in this study it represented only08 This difference could be attributed to the number ofcompounds identified in each study since in this study 63components were identified representing 954 of the totaloil content The essential oil was found to be dominated byoxygenated monoterpenes (Table 1)
Several essential oils components have shown insecti-cidal activity against C maculatus The leaf oil of Cymbo-pogon schoenanthus rich in piperitone gave 90 mortalityafter 24 h at a concentration of 67120583LL [40] Plectranthusgrandifolius essential oil predominantly (E)-myroxide wasshown to be toxic to both adults and eggs of C maculatusLikewise Cinnamomum aromaticum bark oil rich in cis-cinnamaldehyde was insecticidal with an LC
50of 276120583gcm2
after 24 h [41] The essential oil from Cymbopogon giganteusrich in limonene and p-mentha-1(7)8-dien-2-ol demon-strated insecticidal activity against C maculatus and Csubinnotatus [42]The concentrations of themajor terpenoidsconstituents presented here for M lucida essential oil areconsistent with use of this plant as an insecticide and insectrepellent
The insecticidal activity of the M lucida essential oilcould be attributed to those known major components ofoxygenated monoterpenes 18-cineole (434) 120572-terpinylacetate (145) 4-terpinen-4-ol (34) 120572-terpinol (34)and monoterpene hydrocarbons sabinene (82) and 120573-pinene (40) These are compounds whose insecticide andrepellent activities have been reported previously [32 43 44]Nevertheless it also has been shown that minor componentsmay contribute to the biological activity [45] such asMyrcene120572-phellandrene and Camphene
The intensive use of synthetic insecticides not only pro-duces genetic resistance by insect species but also causes seri-ous problems of toxicity to humans and other animal species[6 7] Therefore the essential oil extracted from M lucidarepresents a botanical insecticide source since it showed apotent insecticidal activity comparable to both pesticidesPhostoxin and Primo-ban-20 (Table 2 where confidenceintervals overlap) The lack of a statistical difference betweenM lucida essential oil and two commercial pesticides is apositive result and demonstrates their equivalence thoughwecannot conclude thatM lucida is more effective
The Scientific World Journal 5
0
10
20
30
40
50
60
70
005 01 02 04
Mortality at 24h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(a)
PhostoxinPrimo-ban-20
0102030405060708090
100
005 01 02 04
Mortality at 48h
(120583gmL)
M lucida
(b)
0102030405060708090
100
005 01 02 04
Mortality at 72h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(c)
Figure 1 Effect of theMorinda lucida essential oil and pesticides against Callosobruchus maculatus at different concentrations and exposuretimes
Table 2 Insecticidal activity ofM lucida essential oil against C maculatus
Units LC50 (concentration causing 50 mortality) and LC90 (concentration causing 90 mortality) in units of 120583gmL 95 confidence interval (CI) is shownin parenthesis Insecticidal activity is considered significantly different when the 95 CI fail to overlap aChi-square value of Probit model fit all significant at119875 lt 005 level
6 The Scientific World Journal
The results obtained indicate that M lucida essential oilwas biocidal even at lower concentration and may thereforebe considered to be a useful alternative to synthetic insecti-cides
5 Conclusions
This study demonstrated that essential oil extracted fromMlucida was toxic to pulse beetle C maculatus Therefore itcould be used as an alternative strategy for C maculatuscontrol and as a substitute for synthetic pesticides Howeverfurther studies are necessary to elucidate the mode of actionand their environmental impact and develop formulations toimprove the insecticidal efficacy This would benefit agricul-tural sectors of developing countries as these essential oils arereadily available and biodegradableThe problem of volatilitycan be resolved through a controlled release formulation oftheir active chemical compositions
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Moses S Owolabi and Eduardo Padilla-Camberos con-tributed equally to this work
Acknowledgments
The authors thank Damilola Ajayi for the collection andidentification of the plant sample Mrs Helen Owolabi isgratefully acknowledged for her useful comments
References
[1] C Park S I Kim and Y J Ahn ldquoInsecticidal activity ofasarones identified in Acorus gramineus rhizome against threecoleopteran stored-product insectsrdquo Journal of Stored ProductsResearch vol 39 no 3 pp 333ndash342 2003
[2] A Rahman and F A Talukder ldquoBioefficacy of some plantderivatives that protect grain against the pulse beetle Calloso-bruchus maculatusrdquo Journal of Insect Science vol 6 no 3 pp1ndash10 2006
[3] B Mulatu and T Gebrmedhin ldquoOviposition-deterrent andtoxic effects of various botanicals on the Adzuki bean beetleCallosobruchus chinensis Lrdquo International Journal of TropicalInsect Science vol 20 pp 33ndash38 2000
[4] F A Ajayi and N E S Lale ldquoSeed coat texture host species andtime of application affect the efficacy of essential oils appliedfor the control of Callosobruchus maculatus (F) (ColeopteraBruchidae) in stored pulsesrdquo International Journal of PestManagement vol 47 no 3 pp 161ndash166 2001
[5] L A Tapondjou C Adler H Bouda and D A FontemldquoEfficacy of powder and essential oil fromChenopodium ambro-sioides leaves as post-harvest grain protectants against six-stored product beetlesrdquo Journal of Stored Products Research vol38 no 4 pp 395ndash402 2002
[6] M K Mahmud M M H Khan M Husain M I Alam andM S I Afrad ldquoToxic effects of different plant oils on pulsebeetle Callosobruchus chinensis Linn (Coleoptera Bruchidae)rdquoJournal of the Asiatic Society of Bangladesh Science vol 28 pp11ndash18 2002
[7] FA Lolestani andN Shayesteh ldquoFumigant toxicity ofZiziphoraclinopodioides (Boiss) (Lamiaceae) against adults and eggsof Callosobruchus maculatus (Fab) (Coleoptera Bruchidae)rdquoJournal of Biological Sciences vol 9 no 1 pp 92ndash95 2009
[8] E A Elhag ldquoDeterrent effects of some botanical products onoviposition of the cowpea bruchid Callosobruchus maculatus(F) (Coleoptera Bruchidae)rdquo International Journal of PestManagement vol 46 no 2 pp 109ndash113 2000
[9] C O Adedire and J O Akinneye ldquoBiological activity oftree marigold Tithonia diversifolia on cowpea seed bruchidCallosobruchus maculatus (Coleoptera Bruchidae)rdquo Annals ofApplied Biology vol 144 pp 185ndash189 2004
[10] P Koona and S Dorn ldquoExtracts from Tephrosia vogelii forthe protection of stored legume seeds against damage by threebruchid speciesrdquo Annals of Applied Biology vol 147 no 1 pp43ndash48 2005
[11] D P Papachristos A C Kimbaris N T Papadopoulos and MG Polissiou ldquoToxicity of citrus essential oils against Ceratitiscapitata (Diptera Tephritidae) larvaerdquo Annals of Applied Biol-ogy vol 155 no 3 pp 381ndash389 2009
[12] M Martinez-Velazquez G A Castillo-Herrera R Rosario-Cruz et al ldquoAcaricidal effect and chemical composition ofessential oils extracted from Cuminum cyminum Pimentadioica and Ocimum basilicum against the cattle tick Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo ParasitologyResearch vol 108 no 2 pp 481ndash487 2011
[13] R S Kawuki A Agona P Nampala and E Adipala ldquoA compar-ison of effectiveness of plant-based and synthetic insecticides inthe field management of pod and storage pests of cowpeardquoCropProtection vol 24 no 5 pp 473ndash478 2005
[14] I Mahfuz and M Khalequzzaman ldquoContact and fumiganttoxicity of essential oils against Callosobruchus maculatusrdquoUniversity Journal of Zoology Rajshahi University vol 26 pp63ndash66 2007
[15] I Chiffelle A Huerta M Celis and J E Araya ldquoProximalanalysis and insecticidal effects of extracts from pepper tree(Schinus molle) leaves on elm leaf beetle (Xanthogalerucaluteola) larvaerdquo Industrial Crops and Products vol 43 no 1 pp523ndash528 2013
[16] S M Keita C Vincent J Schmit J T Arnason and ABelanger ldquoEfficacy of essential oil of Ocimum basilicum Land O gratissimum L applied as an insecticidal fumigant andpowder to controlCallosobruchusmaculatus (Fab) [ColeopteraBruchidae]rdquo Journal of Stored Products Research vol 37 no 4pp 339ndash349 2001
[17] M J Pascual-Villalobos and M C Ballesta-Acosta ldquoChemicalvariation in an Ocimum basilicum germplasm collection andactivity of the essential oils on Callosobruchus maculatusrdquoBiochemical Systematics and Ecology vol 31 no 7 pp 673ndash6792003
[18] R Pavela ldquoInsecticidal and repellent activity of selected essen-tial oils against of the pollen beetle Meligethes aeneus (Fabri-cius) adultsrdquo Industrial Crops and Products vol 34 no 1 pp888ndash892 2011
[19] M Martinez-Velazquez R Rosario-Cruz G Castillo-HerreraJ M Flores-Fernandez A H Alvarez and E Lugo-Cervantes
The Scientific World Journal 7
ldquoAcaricidal effect of EssentialOils fromLippia graveolens (Lami-ales Verbenaceae) Rosmarinus officinalis (Lamiales Lami-aceae) and Allium sativum (Liliales Liliaceae) Against Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo Journal ofMedical Entomology vol 48 no 4 pp 822ndash827 2011
[20] S D Karou T Tchacondo D P Ilboudo and J Simpore ldquoSaha-ran Rubiaceae a review of their traditional uses phytochem-istry and biological activitiesrdquo Pakistan Journal of BiologicalSciences vol 14 no 3 pp 149ndash169 2011
[21] J MMakinde and P O Obih ldquoScreening ofMorinda lucida leafextract for antimalarial action on Plasmodium berghei in micerdquoAfrican Journal of Medicine andMedical Sciences vol 17 pp 59ndash63 1985
[22] B Oliver-Bever Medicinal Plants in Tropical West AfricaCambridge University Press Cambridge Mass USA 1986
[23] K M Joppa A Vovor K Eklu-Gadegbeku A Agbonon KAklikokou and M Gbeassor ldquoEffect of Morinda Lucida benth(Rubiaceae) and Newbouldia leavis P beauv (Bignoniaceae) onsickling of red blood cellsrdquoMedecine Tropicale vol 68 no 3 pp251ndash256 2008
[24] K Kovendan K Murugan S P Shanthakumar S Vincentand J Hwang ldquoLarvicidal activity of Morinda citrifolia L(Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012
[25] Z Ilboudo L C B Dabire R C H Nebie et al ldquoBiologicalactivity and persistence of four essential oils towards themain pest of stored cowpeas Callosobruchus maculatus (F)(Coleoptera Bruchidae)rdquo Journal of Stored Products Researchvol 46 no 2 pp 124ndash128 2010
[26] R W D Taylor ldquoPhosphine a major fumigant at riskrdquo Interna-tional Pest Control vol 31 pp 10ndash14 1989
[27] W S Abbott ldquoA method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 pp 265ndash267 1925
[28] D J Finney Probit Analysis Cambridge University Press Cam-bridge UK 3rd edition 1971
[29] S A Bankole ldquoEffect of essential oils from two Nigerianmedicinal plants (Azadirachta indica and Morinda lucida)on growth and aflatoxin B1 production in maize grain by atoxigenicAspergillus flavusrdquoLetters inAppliedMicrobiology vol24 no 3 pp 190ndash192 1997
[30] J S Ashidi P J Houghton P J Hylands and T EfferthldquoEthnobotanical survey and cytotoxicity testing of plants ofSouth-western Nigeria used to treat cancer with isolationof cytotoxic constituents from Cajanus cajan Millsp leavesrdquoJournal of Ethnopharmacology vol 128 no 2 pp 501ndash512 2010
[31] L Lajide C O Adedire W A Muse and S O AgeleldquoInsecticidal activity of Powders of some Nigerian plant againstthe maize weevils (Sitophilus zeamais Mots) in stored maizerdquoEntomology and the Nigerian Economy vol 31 pp 227ndash2351998
[32] T S L Ngamo I Ngatanko M B Ngassoum P M Mapong-mestsem and T Hance ldquoPersistence of insecticidal activities ofcrude essential oils of three aromatic plants towards four majorstored product insect pestsrdquo African Journal of AgriculturalResearch vol 2 pp 173ndash177 2007
[33] M Negahban and S Moharramipour ldquoFumigant toxicityof Eucalyptus intertexta Eucalyptus sargentii and Eucalyp-tus camaldulensis against stored-product beetlesrdquo Journal ofApplied Entomology vol 131 no 4 pp 256ndash261 2007
[34] A F Ndomo L A Tapondjou L T Ngamo and T HanceldquoInsecticidal activities of essential oil of Callistemon viminalisapplied as fumigant and powder against two bruchidsrdquo Journalof Applied Entomology vol 134 no 4 pp 333ndash341 2010
[35] M S Owolabi M O Oladimeji L Lajide G Singh PMarimuthu and V A Isidorov ldquoBioactivity of three plantderived essential oils agianst the maize weevils Sitophilus zea-mais(Motschulsky) and cowpea weevils Callosobruchus macu-latus (Fabricius)rdquo Electronic Journal of Environmental Agricul-tural and Food Chemistry vol 8 no 9 pp 828ndash835 2009
[36] M K Chaubey ldquoFumigant toxicity of essential oils from somecommon spices against pulse beetle Callosobruchus chinensis(Coleoptera Bruchidae)rdquo Journal of Oleo Science vol 57 no 3pp 171ndash179 2008
[37] S Moharramipour A Taghizadeh M H Meshkatalsadat AA Talebi and Y Fathipour ldquoRepellent and fumigant toxicity ofessential oil fromThymus persicus against Tribolium castaneumand Callosobruchus maculatusrdquo Communications in Agricul-tural and Applied Biological Sciences vol 73 no 3 pp 639ndash6422008
[38] S O Okoh O T Asekun O B Familoni and A J AfolayanldquoComposition and antioxidant activities of leaf and root volatileoils ofMorinda lucidardquoNatural Product Communications vol 6no 10 pp 1537ndash1541 2011
[39] K Koumaglo M Gbeassor O Nikabu C de Souza and WWerner ldquoEffects of three compounds extracted from Morindalucida on Plasmodium falciparumrdquo Planta Medica vol 58 no6 pp 533ndash534 1992
[40] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis (Rondani) (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 no 4 pp 363ndash371 2005
[41] R Islam R I Khan S M Al-Reza Y T Jeong C H Song andM Khalequzzaman ldquoChemical composition and insecticidalproperties of Cinnamomum aromaticum (Nees) essential oilagainst the stored product beetleCallosobruchusmaculatus (F)rdquoJournal of the Science of Food and Agriculture vol 89 no 7 pp1241ndash1246 2009
[42] W S Nyamador G K Ketoh K Amevoin Y Nuto H KKoumaglo and I A Glitho ldquoVariation in the susceptibility oftwo Callosobruchus species to essential oilsrdquo Journal of StoredProducts Research vol 46 no 1 pp 48ndash51 2010
[43] H Kouninki T Hance F A Noudjou et al ldquoToxicity ofsome terpenoids of essential oils of Xylopia aethiopica fromCameroon against Sitophilus zeamais Motschulskyrdquo Journal ofApplied Entomology vol 131 no 4 pp 269ndash274 2007
[44] R Waliwitiya M B Isman R S Vernon and A RisemanldquoInsecticidal activity of selected monoterpenoids and rosemaryoil to Agriotes obscurus (Coleoptera Elateridae)rdquo Journal ofEconomic Entomology vol 98 no 5 pp 1560ndash1565 2005
[45] N S Iacobellis P Lo Cantore F Capasso and F SenatoreldquoAntibacterial activity of Cuminum cyminum L and Carumcarvi L essential oilsrdquo Journal of Agricultural and Food Chem-istry vol 53 no 1 pp 57ndash61 2005
have directed the need for effective biodegradable pesticides[8 9]
Because of their high volatility plant extracts or plantcompounds and the use of natural compounds with insec-ticidal activity provide a potential biodegradable alternativeto synthetic pesticides [10ndash12] The contact and fumigantinsecticidal activity of several plant essential oils and theirconstituents have been demonstrated against stored-productpests [13ndash15] a number of bruchid pests [16ndash18] and insectsand mites [19]
Several botanical families for pest control are describedsuch as Meliaceae Rutaceae Asteraceae Labiatae and Mal-vaceae Rubiaceae is a family of 630 genera and about13000 species [20] Morinda lucida (Benth) a member ofRubiaceaefamily is a medicinal plant of about 15m tall with adense crown of slender crooked branches approximately 20ndash30 cm in diameterM lucida commonly known by the Yorubain southwestern part of Nigeria as ldquoOruwordquo is widely dis-tributed in West Africa and is used in African folk medicineto treat several diseases [21] The leaves are bitter and areused by the natives to treat malaria yellow fever jaundicehepatitis eczema edema cough hypertension diabetes andsexual weakness [20 22 23]
In a previous study another species of the same genusM citrifolia showed larvicidal activity against three species ofmosquito vectors [24] and the biofriendly nature ofM lucidasuggested the need to search for new properties of this planttherefore the present study was conducted to determine theinsecticidal activity of the essential oil of M lucida againstC maculatus by dose response bioassays and to characterizeits chemical composition for consideration as an alternativeinsecticide for pulse beetle
2 Materials and Methods
21 Plant Material The aerial parts of the M lucida werecollected fromApomu (7∘19101584048N 4∘10101584047E) which is locatedin Osun State Nigeria and identified at the Herbarium ofthe Botany Department University of Lagos Nigeria wherevoucher specimen number LUH 2637 was kept The sampleswere air-dried pulverized and stored in polythene bag toreduce evaporation of the volatile oil
To extract the essential oil 500 g of pulverized sample wasplaced in a 5 L flask and distilled water was added to coversample Essential oil was obtained by hydrodistillation usingglass modified Clevenger-type apparatus the process ran for4 h at normal atmospheric pressure and at 96-97∘C inside theextractor After the essential oil isolation residual water wasremoved by filtration with anhydrous sodium sulphate andthen the essential oil was stored in an amber vial at 4∘C forfuture analysis
22 Gas Chromatography (GC) Analysis GC analysis wasperformed with a HP-5890 Series II instrument equippedwith a HP-Wax and HP-5 capillary columns (both 30m times025mm ID times 025 120583m film thickness) working with thefollowing temperature program 60∘C for 10min rising at5∘Cmin to 220∘C The injector and detector temperatures
were maintained at 250∘C carrier gas nitrogen (2mLmin)detector dual FID split ratio 1 30 The volume injectedwas 05120583L The relative proportions of the oil constituentswere percentages obtained by FID peak-area normalizationwithout the use of response factor
23 Gas Chromatography-Mass Spectrometry (GC-MS) GC-MS analysis was performed with a Varian CP-3800 gaschromatograph equippedwith aHP-5 capillary column (30mtimes 025mm ID times 025 120583m film thickness) and a Varian Saturn2000 ion trap mass detector Analytical conditions wereinjector and transfer line temperature 220∘C and 240∘Crespectively oven temperature was programmed from 60 to240∘C at 3∘Cmin Carrier gas was helium at 10mLmin flowrate Injection volumewas 02120583L (10hexane solution) usinga 1 30 split ratio The mass spectrometer was operated at 70-eV ionization voltage The acquisition mass range was 30 to300119898119911 at 10 scans
24 Identification of the Volatile Compounds The identifica-tion of the volatile constituents of the essential oil sample wasmade using the following criteria (1) comparison of massspectra with the Wiley library spectra 275 L (RevC0000)electronic database (2) injection of authentic compoundsfrom Sigma-Aldrich with a 95 minimum purity under thesame analytical conditions and (3) comparison of the reten-tion index (RI) obtained by GC-MS with the RI theoreticalin similar phases The molecular weights of all the identifiedsubstances were confirmed by GC-MS using MeOH as CIionizing gas
25 Insect Culture Callosobruchus maculatus were reared oncowpea seeds (Vigna unguiculata) under controlled temper-ature and humidity at 28 plusmn 1∘C 58 plusmn 5 RH (12 h light cycle)and 255 plusmn 1∘C 45plusmn 5RH (12 h dark cycle) Adult insects of8 days old were used for insecticidal test
All experimentswere carried out under the same environ-mental conditions as described above
26 Insecticidal Activity of M lucida Essential Oil The insec-ticidal activity was evaluated as described by Ilboudo et al[25] One hundred and twenty mixed sex adult C maculatus(8 days old) were put into a 500mL glass bottle with 30 g ofcowpea seeds and kept in the laboratory at 28 plusmn 1∘C 58 plusmn 5RH (12 h light cycle) and 255 plusmn 1∘C 45 plusmn 5 RH (12 h darkcycle) for 72 h Concentrations of the M lucida essential oildiluted in acetone were tested on C maculatus (005 010020 and 040 120583gmL) The appropriate concentrations wereapplied to filter paper (Whatman number 1 cut into 7 cmdiameter) and immediately introduced into a glass bottlethat was then hermetically sealed For the control groupthe insects were placed in the glass bottles under the sameconditions but without the essential oil Each concentrationand control was replicated three times Insect mortalitywas determined by observing the recovery of immobilizedinsects in 12 h intervals up to 72 h When no antennal or legmovements were observed insects were considered dead
The Scientific World Journal 3
27 Insecticidal Activity of Commercial Pesticides Two syn-thetic pesticides were purchased from Chemical and Alliedcompany Lagos Nigeria Phostoxin (55 Aluminium phos-phide) and Primo-ban-20 (pirimiphos-methyl emulsifiableconcentrate 50) The first is converted to phosphine gaswhich is taken by the insect through its respiratory systemand the second is able to penetrate the insect cuticle Pelletsof Phostoxin were divided into small pieces and weighedto achieve the appropriate concentrations and were appliedaccording to manufacturerrsquos directions [26] Primo-ban-20was diluted to achieve the desired concentrations from astock solution of 2 120583gmL Both pesticides were evaluated atthe same concentrations as the essential oil in triplicate
28 Mortality Determination Ten pairs of adultCmaculatuswere released each 12 h after exposure of essential oil upto 72 h After a 12 h exposure period the containers wereopened and the dead beetles were removed and recordedKnocked-down adults were regarded as alive if they showedcontinued movement of their appendages by the touch ofa fine brush After observation the containers were tightlyclosed After every 12 h new batches of twenty insects werereleased and the numbers of dead insects were recorded andthe percentage of mortality was calculated and correctedusing Abbottrsquos formula [27]
29 Data Analysis Average percentage insect mortality wascalculated from three replicates Dosage-mortality responseswere used to estimate LC
50(concentration causing 50
mortality) and LC90
(concentration causing 90 mortality)values at 24 and 48 h with Probit analysis [28] using Stat-graphics 51 software Insecticidal activity was consideredsignificantly different when the 95 CI (confidence interval)fail to overlap
3 Results
31 Chemical Composition of Morinda lucida The chemicalcomposition analysis by GC-MS identified 63 components ofM lucida essential oil containing mainly 18-cineole (434)followed by 120572-terpinyl acetate (145) sabinene (82) and120573-pinene (40 Table 1)
32 Mortality of Callosobruchus maculatus The aerial partsof the M lucida distillate yielded a 048 (vw) essentialoil The insecticidal bioassay of the essential oil of M lucidaagainstC maculatus (Figure 1) showed that the activity of theoil was both dose dependent and exposure dependent At adose of 005120583gmL the essential oil produced 594 mortal-ity after 72 h (Figure 1(c)) The essential oil produced 333567 and 899 mortality after 24 48 and 72 h at a dose of010 120583gmL respectively while a dose of 020120583gmL yieldedmortality of 467 833 and 100 respectively over thesame time duration (Figure 1) The highest concentration of040 120583gmL produced a mortality of 90 and 100 after 48and 72 h respectively (Figures 1(b) and 1(c))
The synthetic pesticide Primo-ban-20 at 020120583gmL hada lower mortality (367) at 24 h while Phostoxin had a
Table 1 Retention index and relative composition of major chemi-cal components ofM lucida essential oil
Component KIa 120573-Caryophyllene 1421 Tr119901-Cymen-7-ol-acetate 1423 Tr(119864)-Cinnamyl acetate 1450 Tr(119864)-Methyl isoeugenol 1495 TrBicyclogermacrene 1497 Tr120572-Bulnesene 1506 Tr120573-Sesquiphellandrene 1526 TrElemol 1554 TrElemicin 1557 TrSpathulenol 1578 04Caryophyllene oxide 1583 05g-Eudesmol 1635 Tr120573-Eudesmol 1651 02120572-Eudesmol 1655 TrIntermedeol 1667 TrTotal identified 9540Major components are shown in boldlowastThese chemical components were identified by authentic compound injec-tionaKI Kovats retention indicesTr = trace amount lt01
467 and 867mortality after 24 h and 48 h at 020 120583gmLrespectively (Figures 1(a) and 1(b)) Both M lucida essen-tial oil and pesticides reached 100 mortality at 72 h at020120583gmL (Figure 1(c)) The negative control showed noactivity except at 72 h (17)
33 Determination of Insecticidal Activity Probit analysisshowed that M lucida essential oil after 24 h presentedan insecticidal activity (LC
90= 0629120583gmL) similar to
Phostoxin (LC90
= 0652120583gmL) and Primo-ban-20 (LC90
= 0726 120583gmL) At 24 h essential oil of M lucida (LC50
= 0298 120583gmL) and Phostoxin (LC50
= 0295 120583gmL) weremore efficient than Primo-ban-20 (LC
50= 0393 120583gmL) At
48 h after exposure there was no significant difference inLC50
between M lucida essential oil compared to Primo-ban-20 and Phostoxin (Table 2 where all confidence intervalsoverlap)
4 Discussion
Recently biological activity has been demonstrated for theM lucida essential oil such as the complete suppression ofaflatoxin synthesis in maize [29] cytotoxic activity on cancercell lines [30] and antioxidant and anti-inflammatory activity[20] This study demonstrated a potent insecticidal activityfor M lucida showing 100 mortality on C maculatus atconcentrations of 020 and 040120583gmL after 72 h (Figure 1)
Our results for M lucida essential oil are consistent withother reports of essential oils that exhibited insecticidal andrepellency activity against C maculatus such as Eucalyp-tus intertexta Callistemon viminalis Ocimum americanum
Hyptis suaveolens Hyptis spicigera Lippia multiflora Cym-bopogon citratus and Lippia rugosa [25 31ndash35] Chaubey[36] evaluated the insecticidal oviposition egg hatchingand developmental inhibitory activities of seven differentessential oils against Callosobruchus chinensis and found thatNigella sativa was the most effective at all stages MoreoverKeita et al [16] found that essential oils ofOcimum basilicumand O gratissimum at a dose of 25 120583gmL after 12 h offumigation produced 80 and 70 mortality respectivelySimilarly Moharramipour et al [37] have shown a repellencyof 824 of the Thymus persicus essential oil against Cmaculatus
The chemical components of this essential oil have beenpreviously reported [38 39] although there were differencesin the ratio of their chemicals constituents Okoh et al [38]identified 50 compounds where 120572-terpinene was the majorconstituent with 178 while in this study it represented only08 This difference could be attributed to the number ofcompounds identified in each study since in this study 63components were identified representing 954 of the totaloil content The essential oil was found to be dominated byoxygenated monoterpenes (Table 1)
Several essential oils components have shown insecti-cidal activity against C maculatus The leaf oil of Cymbo-pogon schoenanthus rich in piperitone gave 90 mortalityafter 24 h at a concentration of 67120583LL [40] Plectranthusgrandifolius essential oil predominantly (E)-myroxide wasshown to be toxic to both adults and eggs of C maculatusLikewise Cinnamomum aromaticum bark oil rich in cis-cinnamaldehyde was insecticidal with an LC
50of 276120583gcm2
after 24 h [41] The essential oil from Cymbopogon giganteusrich in limonene and p-mentha-1(7)8-dien-2-ol demon-strated insecticidal activity against C maculatus and Csubinnotatus [42]The concentrations of themajor terpenoidsconstituents presented here for M lucida essential oil areconsistent with use of this plant as an insecticide and insectrepellent
The insecticidal activity of the M lucida essential oilcould be attributed to those known major components ofoxygenated monoterpenes 18-cineole (434) 120572-terpinylacetate (145) 4-terpinen-4-ol (34) 120572-terpinol (34)and monoterpene hydrocarbons sabinene (82) and 120573-pinene (40) These are compounds whose insecticide andrepellent activities have been reported previously [32 43 44]Nevertheless it also has been shown that minor componentsmay contribute to the biological activity [45] such asMyrcene120572-phellandrene and Camphene
The intensive use of synthetic insecticides not only pro-duces genetic resistance by insect species but also causes seri-ous problems of toxicity to humans and other animal species[6 7] Therefore the essential oil extracted from M lucidarepresents a botanical insecticide source since it showed apotent insecticidal activity comparable to both pesticidesPhostoxin and Primo-ban-20 (Table 2 where confidenceintervals overlap) The lack of a statistical difference betweenM lucida essential oil and two commercial pesticides is apositive result and demonstrates their equivalence thoughwecannot conclude thatM lucida is more effective
The Scientific World Journal 5
0
10
20
30
40
50
60
70
005 01 02 04
Mortality at 24h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(a)
PhostoxinPrimo-ban-20
0102030405060708090
100
005 01 02 04
Mortality at 48h
(120583gmL)
M lucida
(b)
0102030405060708090
100
005 01 02 04
Mortality at 72h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(c)
Figure 1 Effect of theMorinda lucida essential oil and pesticides against Callosobruchus maculatus at different concentrations and exposuretimes
Table 2 Insecticidal activity ofM lucida essential oil against C maculatus
Units LC50 (concentration causing 50 mortality) and LC90 (concentration causing 90 mortality) in units of 120583gmL 95 confidence interval (CI) is shownin parenthesis Insecticidal activity is considered significantly different when the 95 CI fail to overlap aChi-square value of Probit model fit all significant at119875 lt 005 level
6 The Scientific World Journal
The results obtained indicate that M lucida essential oilwas biocidal even at lower concentration and may thereforebe considered to be a useful alternative to synthetic insecti-cides
5 Conclusions
This study demonstrated that essential oil extracted fromMlucida was toxic to pulse beetle C maculatus Therefore itcould be used as an alternative strategy for C maculatuscontrol and as a substitute for synthetic pesticides Howeverfurther studies are necessary to elucidate the mode of actionand their environmental impact and develop formulations toimprove the insecticidal efficacy This would benefit agricul-tural sectors of developing countries as these essential oils arereadily available and biodegradableThe problem of volatilitycan be resolved through a controlled release formulation oftheir active chemical compositions
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Moses S Owolabi and Eduardo Padilla-Camberos con-tributed equally to this work
Acknowledgments
The authors thank Damilola Ajayi for the collection andidentification of the plant sample Mrs Helen Owolabi isgratefully acknowledged for her useful comments
References
[1] C Park S I Kim and Y J Ahn ldquoInsecticidal activity ofasarones identified in Acorus gramineus rhizome against threecoleopteran stored-product insectsrdquo Journal of Stored ProductsResearch vol 39 no 3 pp 333ndash342 2003
[2] A Rahman and F A Talukder ldquoBioefficacy of some plantderivatives that protect grain against the pulse beetle Calloso-bruchus maculatusrdquo Journal of Insect Science vol 6 no 3 pp1ndash10 2006
[3] B Mulatu and T Gebrmedhin ldquoOviposition-deterrent andtoxic effects of various botanicals on the Adzuki bean beetleCallosobruchus chinensis Lrdquo International Journal of TropicalInsect Science vol 20 pp 33ndash38 2000
[4] F A Ajayi and N E S Lale ldquoSeed coat texture host species andtime of application affect the efficacy of essential oils appliedfor the control of Callosobruchus maculatus (F) (ColeopteraBruchidae) in stored pulsesrdquo International Journal of PestManagement vol 47 no 3 pp 161ndash166 2001
[5] L A Tapondjou C Adler H Bouda and D A FontemldquoEfficacy of powder and essential oil fromChenopodium ambro-sioides leaves as post-harvest grain protectants against six-stored product beetlesrdquo Journal of Stored Products Research vol38 no 4 pp 395ndash402 2002
[6] M K Mahmud M M H Khan M Husain M I Alam andM S I Afrad ldquoToxic effects of different plant oils on pulsebeetle Callosobruchus chinensis Linn (Coleoptera Bruchidae)rdquoJournal of the Asiatic Society of Bangladesh Science vol 28 pp11ndash18 2002
[7] FA Lolestani andN Shayesteh ldquoFumigant toxicity ofZiziphoraclinopodioides (Boiss) (Lamiaceae) against adults and eggsof Callosobruchus maculatus (Fab) (Coleoptera Bruchidae)rdquoJournal of Biological Sciences vol 9 no 1 pp 92ndash95 2009
[8] E A Elhag ldquoDeterrent effects of some botanical products onoviposition of the cowpea bruchid Callosobruchus maculatus(F) (Coleoptera Bruchidae)rdquo International Journal of PestManagement vol 46 no 2 pp 109ndash113 2000
[9] C O Adedire and J O Akinneye ldquoBiological activity oftree marigold Tithonia diversifolia on cowpea seed bruchidCallosobruchus maculatus (Coleoptera Bruchidae)rdquo Annals ofApplied Biology vol 144 pp 185ndash189 2004
[10] P Koona and S Dorn ldquoExtracts from Tephrosia vogelii forthe protection of stored legume seeds against damage by threebruchid speciesrdquo Annals of Applied Biology vol 147 no 1 pp43ndash48 2005
[11] D P Papachristos A C Kimbaris N T Papadopoulos and MG Polissiou ldquoToxicity of citrus essential oils against Ceratitiscapitata (Diptera Tephritidae) larvaerdquo Annals of Applied Biol-ogy vol 155 no 3 pp 381ndash389 2009
[12] M Martinez-Velazquez G A Castillo-Herrera R Rosario-Cruz et al ldquoAcaricidal effect and chemical composition ofessential oils extracted from Cuminum cyminum Pimentadioica and Ocimum basilicum against the cattle tick Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo ParasitologyResearch vol 108 no 2 pp 481ndash487 2011
[13] R S Kawuki A Agona P Nampala and E Adipala ldquoA compar-ison of effectiveness of plant-based and synthetic insecticides inthe field management of pod and storage pests of cowpeardquoCropProtection vol 24 no 5 pp 473ndash478 2005
[14] I Mahfuz and M Khalequzzaman ldquoContact and fumiganttoxicity of essential oils against Callosobruchus maculatusrdquoUniversity Journal of Zoology Rajshahi University vol 26 pp63ndash66 2007
[15] I Chiffelle A Huerta M Celis and J E Araya ldquoProximalanalysis and insecticidal effects of extracts from pepper tree(Schinus molle) leaves on elm leaf beetle (Xanthogalerucaluteola) larvaerdquo Industrial Crops and Products vol 43 no 1 pp523ndash528 2013
[16] S M Keita C Vincent J Schmit J T Arnason and ABelanger ldquoEfficacy of essential oil of Ocimum basilicum Land O gratissimum L applied as an insecticidal fumigant andpowder to controlCallosobruchusmaculatus (Fab) [ColeopteraBruchidae]rdquo Journal of Stored Products Research vol 37 no 4pp 339ndash349 2001
[17] M J Pascual-Villalobos and M C Ballesta-Acosta ldquoChemicalvariation in an Ocimum basilicum germplasm collection andactivity of the essential oils on Callosobruchus maculatusrdquoBiochemical Systematics and Ecology vol 31 no 7 pp 673ndash6792003
[18] R Pavela ldquoInsecticidal and repellent activity of selected essen-tial oils against of the pollen beetle Meligethes aeneus (Fabri-cius) adultsrdquo Industrial Crops and Products vol 34 no 1 pp888ndash892 2011
[19] M Martinez-Velazquez R Rosario-Cruz G Castillo-HerreraJ M Flores-Fernandez A H Alvarez and E Lugo-Cervantes
The Scientific World Journal 7
ldquoAcaricidal effect of EssentialOils fromLippia graveolens (Lami-ales Verbenaceae) Rosmarinus officinalis (Lamiales Lami-aceae) and Allium sativum (Liliales Liliaceae) Against Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo Journal ofMedical Entomology vol 48 no 4 pp 822ndash827 2011
[20] S D Karou T Tchacondo D P Ilboudo and J Simpore ldquoSaha-ran Rubiaceae a review of their traditional uses phytochem-istry and biological activitiesrdquo Pakistan Journal of BiologicalSciences vol 14 no 3 pp 149ndash169 2011
[21] J MMakinde and P O Obih ldquoScreening ofMorinda lucida leafextract for antimalarial action on Plasmodium berghei in micerdquoAfrican Journal of Medicine andMedical Sciences vol 17 pp 59ndash63 1985
[22] B Oliver-Bever Medicinal Plants in Tropical West AfricaCambridge University Press Cambridge Mass USA 1986
[23] K M Joppa A Vovor K Eklu-Gadegbeku A Agbonon KAklikokou and M Gbeassor ldquoEffect of Morinda Lucida benth(Rubiaceae) and Newbouldia leavis P beauv (Bignoniaceae) onsickling of red blood cellsrdquoMedecine Tropicale vol 68 no 3 pp251ndash256 2008
[24] K Kovendan K Murugan S P Shanthakumar S Vincentand J Hwang ldquoLarvicidal activity of Morinda citrifolia L(Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012
[25] Z Ilboudo L C B Dabire R C H Nebie et al ldquoBiologicalactivity and persistence of four essential oils towards themain pest of stored cowpeas Callosobruchus maculatus (F)(Coleoptera Bruchidae)rdquo Journal of Stored Products Researchvol 46 no 2 pp 124ndash128 2010
[26] R W D Taylor ldquoPhosphine a major fumigant at riskrdquo Interna-tional Pest Control vol 31 pp 10ndash14 1989
[27] W S Abbott ldquoA method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 pp 265ndash267 1925
[28] D J Finney Probit Analysis Cambridge University Press Cam-bridge UK 3rd edition 1971
[29] S A Bankole ldquoEffect of essential oils from two Nigerianmedicinal plants (Azadirachta indica and Morinda lucida)on growth and aflatoxin B1 production in maize grain by atoxigenicAspergillus flavusrdquoLetters inAppliedMicrobiology vol24 no 3 pp 190ndash192 1997
[30] J S Ashidi P J Houghton P J Hylands and T EfferthldquoEthnobotanical survey and cytotoxicity testing of plants ofSouth-western Nigeria used to treat cancer with isolationof cytotoxic constituents from Cajanus cajan Millsp leavesrdquoJournal of Ethnopharmacology vol 128 no 2 pp 501ndash512 2010
[31] L Lajide C O Adedire W A Muse and S O AgeleldquoInsecticidal activity of Powders of some Nigerian plant againstthe maize weevils (Sitophilus zeamais Mots) in stored maizerdquoEntomology and the Nigerian Economy vol 31 pp 227ndash2351998
[32] T S L Ngamo I Ngatanko M B Ngassoum P M Mapong-mestsem and T Hance ldquoPersistence of insecticidal activities ofcrude essential oils of three aromatic plants towards four majorstored product insect pestsrdquo African Journal of AgriculturalResearch vol 2 pp 173ndash177 2007
[33] M Negahban and S Moharramipour ldquoFumigant toxicityof Eucalyptus intertexta Eucalyptus sargentii and Eucalyp-tus camaldulensis against stored-product beetlesrdquo Journal ofApplied Entomology vol 131 no 4 pp 256ndash261 2007
[34] A F Ndomo L A Tapondjou L T Ngamo and T HanceldquoInsecticidal activities of essential oil of Callistemon viminalisapplied as fumigant and powder against two bruchidsrdquo Journalof Applied Entomology vol 134 no 4 pp 333ndash341 2010
[35] M S Owolabi M O Oladimeji L Lajide G Singh PMarimuthu and V A Isidorov ldquoBioactivity of three plantderived essential oils agianst the maize weevils Sitophilus zea-mais(Motschulsky) and cowpea weevils Callosobruchus macu-latus (Fabricius)rdquo Electronic Journal of Environmental Agricul-tural and Food Chemistry vol 8 no 9 pp 828ndash835 2009
[36] M K Chaubey ldquoFumigant toxicity of essential oils from somecommon spices against pulse beetle Callosobruchus chinensis(Coleoptera Bruchidae)rdquo Journal of Oleo Science vol 57 no 3pp 171ndash179 2008
[37] S Moharramipour A Taghizadeh M H Meshkatalsadat AA Talebi and Y Fathipour ldquoRepellent and fumigant toxicity ofessential oil fromThymus persicus against Tribolium castaneumand Callosobruchus maculatusrdquo Communications in Agricul-tural and Applied Biological Sciences vol 73 no 3 pp 639ndash6422008
[38] S O Okoh O T Asekun O B Familoni and A J AfolayanldquoComposition and antioxidant activities of leaf and root volatileoils ofMorinda lucidardquoNatural Product Communications vol 6no 10 pp 1537ndash1541 2011
[39] K Koumaglo M Gbeassor O Nikabu C de Souza and WWerner ldquoEffects of three compounds extracted from Morindalucida on Plasmodium falciparumrdquo Planta Medica vol 58 no6 pp 533ndash534 1992
[40] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis (Rondani) (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 no 4 pp 363ndash371 2005
[41] R Islam R I Khan S M Al-Reza Y T Jeong C H Song andM Khalequzzaman ldquoChemical composition and insecticidalproperties of Cinnamomum aromaticum (Nees) essential oilagainst the stored product beetleCallosobruchusmaculatus (F)rdquoJournal of the Science of Food and Agriculture vol 89 no 7 pp1241ndash1246 2009
[42] W S Nyamador G K Ketoh K Amevoin Y Nuto H KKoumaglo and I A Glitho ldquoVariation in the susceptibility oftwo Callosobruchus species to essential oilsrdquo Journal of StoredProducts Research vol 46 no 1 pp 48ndash51 2010
[43] H Kouninki T Hance F A Noudjou et al ldquoToxicity ofsome terpenoids of essential oils of Xylopia aethiopica fromCameroon against Sitophilus zeamais Motschulskyrdquo Journal ofApplied Entomology vol 131 no 4 pp 269ndash274 2007
[44] R Waliwitiya M B Isman R S Vernon and A RisemanldquoInsecticidal activity of selected monoterpenoids and rosemaryoil to Agriotes obscurus (Coleoptera Elateridae)rdquo Journal ofEconomic Entomology vol 98 no 5 pp 1560ndash1565 2005
[45] N S Iacobellis P Lo Cantore F Capasso and F SenatoreldquoAntibacterial activity of Cuminum cyminum L and Carumcarvi L essential oilsrdquo Journal of Agricultural and Food Chem-istry vol 53 no 1 pp 57ndash61 2005
27 Insecticidal Activity of Commercial Pesticides Two syn-thetic pesticides were purchased from Chemical and Alliedcompany Lagos Nigeria Phostoxin (55 Aluminium phos-phide) and Primo-ban-20 (pirimiphos-methyl emulsifiableconcentrate 50) The first is converted to phosphine gaswhich is taken by the insect through its respiratory systemand the second is able to penetrate the insect cuticle Pelletsof Phostoxin were divided into small pieces and weighedto achieve the appropriate concentrations and were appliedaccording to manufacturerrsquos directions [26] Primo-ban-20was diluted to achieve the desired concentrations from astock solution of 2 120583gmL Both pesticides were evaluated atthe same concentrations as the essential oil in triplicate
28 Mortality Determination Ten pairs of adultCmaculatuswere released each 12 h after exposure of essential oil upto 72 h After a 12 h exposure period the containers wereopened and the dead beetles were removed and recordedKnocked-down adults were regarded as alive if they showedcontinued movement of their appendages by the touch ofa fine brush After observation the containers were tightlyclosed After every 12 h new batches of twenty insects werereleased and the numbers of dead insects were recorded andthe percentage of mortality was calculated and correctedusing Abbottrsquos formula [27]
29 Data Analysis Average percentage insect mortality wascalculated from three replicates Dosage-mortality responseswere used to estimate LC
50(concentration causing 50
mortality) and LC90
(concentration causing 90 mortality)values at 24 and 48 h with Probit analysis [28] using Stat-graphics 51 software Insecticidal activity was consideredsignificantly different when the 95 CI (confidence interval)fail to overlap
3 Results
31 Chemical Composition of Morinda lucida The chemicalcomposition analysis by GC-MS identified 63 components ofM lucida essential oil containing mainly 18-cineole (434)followed by 120572-terpinyl acetate (145) sabinene (82) and120573-pinene (40 Table 1)
32 Mortality of Callosobruchus maculatus The aerial partsof the M lucida distillate yielded a 048 (vw) essentialoil The insecticidal bioassay of the essential oil of M lucidaagainstC maculatus (Figure 1) showed that the activity of theoil was both dose dependent and exposure dependent At adose of 005120583gmL the essential oil produced 594 mortal-ity after 72 h (Figure 1(c)) The essential oil produced 333567 and 899 mortality after 24 48 and 72 h at a dose of010 120583gmL respectively while a dose of 020120583gmL yieldedmortality of 467 833 and 100 respectively over thesame time duration (Figure 1) The highest concentration of040 120583gmL produced a mortality of 90 and 100 after 48and 72 h respectively (Figures 1(b) and 1(c))
The synthetic pesticide Primo-ban-20 at 020120583gmL hada lower mortality (367) at 24 h while Phostoxin had a
Table 1 Retention index and relative composition of major chemi-cal components ofM lucida essential oil
Component KIa 120573-Caryophyllene 1421 Tr119901-Cymen-7-ol-acetate 1423 Tr(119864)-Cinnamyl acetate 1450 Tr(119864)-Methyl isoeugenol 1495 TrBicyclogermacrene 1497 Tr120572-Bulnesene 1506 Tr120573-Sesquiphellandrene 1526 TrElemol 1554 TrElemicin 1557 TrSpathulenol 1578 04Caryophyllene oxide 1583 05g-Eudesmol 1635 Tr120573-Eudesmol 1651 02120572-Eudesmol 1655 TrIntermedeol 1667 TrTotal identified 9540Major components are shown in boldlowastThese chemical components were identified by authentic compound injec-tionaKI Kovats retention indicesTr = trace amount lt01
467 and 867mortality after 24 h and 48 h at 020 120583gmLrespectively (Figures 1(a) and 1(b)) Both M lucida essen-tial oil and pesticides reached 100 mortality at 72 h at020120583gmL (Figure 1(c)) The negative control showed noactivity except at 72 h (17)
33 Determination of Insecticidal Activity Probit analysisshowed that M lucida essential oil after 24 h presentedan insecticidal activity (LC
90= 0629120583gmL) similar to
Phostoxin (LC90
= 0652120583gmL) and Primo-ban-20 (LC90
= 0726 120583gmL) At 24 h essential oil of M lucida (LC50
= 0298 120583gmL) and Phostoxin (LC50
= 0295 120583gmL) weremore efficient than Primo-ban-20 (LC
50= 0393 120583gmL) At
48 h after exposure there was no significant difference inLC50
between M lucida essential oil compared to Primo-ban-20 and Phostoxin (Table 2 where all confidence intervalsoverlap)
4 Discussion
Recently biological activity has been demonstrated for theM lucida essential oil such as the complete suppression ofaflatoxin synthesis in maize [29] cytotoxic activity on cancercell lines [30] and antioxidant and anti-inflammatory activity[20] This study demonstrated a potent insecticidal activityfor M lucida showing 100 mortality on C maculatus atconcentrations of 020 and 040120583gmL after 72 h (Figure 1)
Our results for M lucida essential oil are consistent withother reports of essential oils that exhibited insecticidal andrepellency activity against C maculatus such as Eucalyp-tus intertexta Callistemon viminalis Ocimum americanum
Hyptis suaveolens Hyptis spicigera Lippia multiflora Cym-bopogon citratus and Lippia rugosa [25 31ndash35] Chaubey[36] evaluated the insecticidal oviposition egg hatchingand developmental inhibitory activities of seven differentessential oils against Callosobruchus chinensis and found thatNigella sativa was the most effective at all stages MoreoverKeita et al [16] found that essential oils ofOcimum basilicumand O gratissimum at a dose of 25 120583gmL after 12 h offumigation produced 80 and 70 mortality respectivelySimilarly Moharramipour et al [37] have shown a repellencyof 824 of the Thymus persicus essential oil against Cmaculatus
The chemical components of this essential oil have beenpreviously reported [38 39] although there were differencesin the ratio of their chemicals constituents Okoh et al [38]identified 50 compounds where 120572-terpinene was the majorconstituent with 178 while in this study it represented only08 This difference could be attributed to the number ofcompounds identified in each study since in this study 63components were identified representing 954 of the totaloil content The essential oil was found to be dominated byoxygenated monoterpenes (Table 1)
Several essential oils components have shown insecti-cidal activity against C maculatus The leaf oil of Cymbo-pogon schoenanthus rich in piperitone gave 90 mortalityafter 24 h at a concentration of 67120583LL [40] Plectranthusgrandifolius essential oil predominantly (E)-myroxide wasshown to be toxic to both adults and eggs of C maculatusLikewise Cinnamomum aromaticum bark oil rich in cis-cinnamaldehyde was insecticidal with an LC
50of 276120583gcm2
after 24 h [41] The essential oil from Cymbopogon giganteusrich in limonene and p-mentha-1(7)8-dien-2-ol demon-strated insecticidal activity against C maculatus and Csubinnotatus [42]The concentrations of themajor terpenoidsconstituents presented here for M lucida essential oil areconsistent with use of this plant as an insecticide and insectrepellent
The insecticidal activity of the M lucida essential oilcould be attributed to those known major components ofoxygenated monoterpenes 18-cineole (434) 120572-terpinylacetate (145) 4-terpinen-4-ol (34) 120572-terpinol (34)and monoterpene hydrocarbons sabinene (82) and 120573-pinene (40) These are compounds whose insecticide andrepellent activities have been reported previously [32 43 44]Nevertheless it also has been shown that minor componentsmay contribute to the biological activity [45] such asMyrcene120572-phellandrene and Camphene
The intensive use of synthetic insecticides not only pro-duces genetic resistance by insect species but also causes seri-ous problems of toxicity to humans and other animal species[6 7] Therefore the essential oil extracted from M lucidarepresents a botanical insecticide source since it showed apotent insecticidal activity comparable to both pesticidesPhostoxin and Primo-ban-20 (Table 2 where confidenceintervals overlap) The lack of a statistical difference betweenM lucida essential oil and two commercial pesticides is apositive result and demonstrates their equivalence thoughwecannot conclude thatM lucida is more effective
The Scientific World Journal 5
0
10
20
30
40
50
60
70
005 01 02 04
Mortality at 24h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(a)
PhostoxinPrimo-ban-20
0102030405060708090
100
005 01 02 04
Mortality at 48h
(120583gmL)
M lucida
(b)
0102030405060708090
100
005 01 02 04
Mortality at 72h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(c)
Figure 1 Effect of theMorinda lucida essential oil and pesticides against Callosobruchus maculatus at different concentrations and exposuretimes
Table 2 Insecticidal activity ofM lucida essential oil against C maculatus
Units LC50 (concentration causing 50 mortality) and LC90 (concentration causing 90 mortality) in units of 120583gmL 95 confidence interval (CI) is shownin parenthesis Insecticidal activity is considered significantly different when the 95 CI fail to overlap aChi-square value of Probit model fit all significant at119875 lt 005 level
6 The Scientific World Journal
The results obtained indicate that M lucida essential oilwas biocidal even at lower concentration and may thereforebe considered to be a useful alternative to synthetic insecti-cides
5 Conclusions
This study demonstrated that essential oil extracted fromMlucida was toxic to pulse beetle C maculatus Therefore itcould be used as an alternative strategy for C maculatuscontrol and as a substitute for synthetic pesticides Howeverfurther studies are necessary to elucidate the mode of actionand their environmental impact and develop formulations toimprove the insecticidal efficacy This would benefit agricul-tural sectors of developing countries as these essential oils arereadily available and biodegradableThe problem of volatilitycan be resolved through a controlled release formulation oftheir active chemical compositions
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Moses S Owolabi and Eduardo Padilla-Camberos con-tributed equally to this work
Acknowledgments
The authors thank Damilola Ajayi for the collection andidentification of the plant sample Mrs Helen Owolabi isgratefully acknowledged for her useful comments
References
[1] C Park S I Kim and Y J Ahn ldquoInsecticidal activity ofasarones identified in Acorus gramineus rhizome against threecoleopteran stored-product insectsrdquo Journal of Stored ProductsResearch vol 39 no 3 pp 333ndash342 2003
[2] A Rahman and F A Talukder ldquoBioefficacy of some plantderivatives that protect grain against the pulse beetle Calloso-bruchus maculatusrdquo Journal of Insect Science vol 6 no 3 pp1ndash10 2006
[3] B Mulatu and T Gebrmedhin ldquoOviposition-deterrent andtoxic effects of various botanicals on the Adzuki bean beetleCallosobruchus chinensis Lrdquo International Journal of TropicalInsect Science vol 20 pp 33ndash38 2000
[4] F A Ajayi and N E S Lale ldquoSeed coat texture host species andtime of application affect the efficacy of essential oils appliedfor the control of Callosobruchus maculatus (F) (ColeopteraBruchidae) in stored pulsesrdquo International Journal of PestManagement vol 47 no 3 pp 161ndash166 2001
[5] L A Tapondjou C Adler H Bouda and D A FontemldquoEfficacy of powder and essential oil fromChenopodium ambro-sioides leaves as post-harvest grain protectants against six-stored product beetlesrdquo Journal of Stored Products Research vol38 no 4 pp 395ndash402 2002
[6] M K Mahmud M M H Khan M Husain M I Alam andM S I Afrad ldquoToxic effects of different plant oils on pulsebeetle Callosobruchus chinensis Linn (Coleoptera Bruchidae)rdquoJournal of the Asiatic Society of Bangladesh Science vol 28 pp11ndash18 2002
[7] FA Lolestani andN Shayesteh ldquoFumigant toxicity ofZiziphoraclinopodioides (Boiss) (Lamiaceae) against adults and eggsof Callosobruchus maculatus (Fab) (Coleoptera Bruchidae)rdquoJournal of Biological Sciences vol 9 no 1 pp 92ndash95 2009
[8] E A Elhag ldquoDeterrent effects of some botanical products onoviposition of the cowpea bruchid Callosobruchus maculatus(F) (Coleoptera Bruchidae)rdquo International Journal of PestManagement vol 46 no 2 pp 109ndash113 2000
[9] C O Adedire and J O Akinneye ldquoBiological activity oftree marigold Tithonia diversifolia on cowpea seed bruchidCallosobruchus maculatus (Coleoptera Bruchidae)rdquo Annals ofApplied Biology vol 144 pp 185ndash189 2004
[10] P Koona and S Dorn ldquoExtracts from Tephrosia vogelii forthe protection of stored legume seeds against damage by threebruchid speciesrdquo Annals of Applied Biology vol 147 no 1 pp43ndash48 2005
[11] D P Papachristos A C Kimbaris N T Papadopoulos and MG Polissiou ldquoToxicity of citrus essential oils against Ceratitiscapitata (Diptera Tephritidae) larvaerdquo Annals of Applied Biol-ogy vol 155 no 3 pp 381ndash389 2009
[12] M Martinez-Velazquez G A Castillo-Herrera R Rosario-Cruz et al ldquoAcaricidal effect and chemical composition ofessential oils extracted from Cuminum cyminum Pimentadioica and Ocimum basilicum against the cattle tick Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo ParasitologyResearch vol 108 no 2 pp 481ndash487 2011
[13] R S Kawuki A Agona P Nampala and E Adipala ldquoA compar-ison of effectiveness of plant-based and synthetic insecticides inthe field management of pod and storage pests of cowpeardquoCropProtection vol 24 no 5 pp 473ndash478 2005
[14] I Mahfuz and M Khalequzzaman ldquoContact and fumiganttoxicity of essential oils against Callosobruchus maculatusrdquoUniversity Journal of Zoology Rajshahi University vol 26 pp63ndash66 2007
[15] I Chiffelle A Huerta M Celis and J E Araya ldquoProximalanalysis and insecticidal effects of extracts from pepper tree(Schinus molle) leaves on elm leaf beetle (Xanthogalerucaluteola) larvaerdquo Industrial Crops and Products vol 43 no 1 pp523ndash528 2013
[16] S M Keita C Vincent J Schmit J T Arnason and ABelanger ldquoEfficacy of essential oil of Ocimum basilicum Land O gratissimum L applied as an insecticidal fumigant andpowder to controlCallosobruchusmaculatus (Fab) [ColeopteraBruchidae]rdquo Journal of Stored Products Research vol 37 no 4pp 339ndash349 2001
[17] M J Pascual-Villalobos and M C Ballesta-Acosta ldquoChemicalvariation in an Ocimum basilicum germplasm collection andactivity of the essential oils on Callosobruchus maculatusrdquoBiochemical Systematics and Ecology vol 31 no 7 pp 673ndash6792003
[18] R Pavela ldquoInsecticidal and repellent activity of selected essen-tial oils against of the pollen beetle Meligethes aeneus (Fabri-cius) adultsrdquo Industrial Crops and Products vol 34 no 1 pp888ndash892 2011
[19] M Martinez-Velazquez R Rosario-Cruz G Castillo-HerreraJ M Flores-Fernandez A H Alvarez and E Lugo-Cervantes
The Scientific World Journal 7
ldquoAcaricidal effect of EssentialOils fromLippia graveolens (Lami-ales Verbenaceae) Rosmarinus officinalis (Lamiales Lami-aceae) and Allium sativum (Liliales Liliaceae) Against Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo Journal ofMedical Entomology vol 48 no 4 pp 822ndash827 2011
[20] S D Karou T Tchacondo D P Ilboudo and J Simpore ldquoSaha-ran Rubiaceae a review of their traditional uses phytochem-istry and biological activitiesrdquo Pakistan Journal of BiologicalSciences vol 14 no 3 pp 149ndash169 2011
[21] J MMakinde and P O Obih ldquoScreening ofMorinda lucida leafextract for antimalarial action on Plasmodium berghei in micerdquoAfrican Journal of Medicine andMedical Sciences vol 17 pp 59ndash63 1985
[22] B Oliver-Bever Medicinal Plants in Tropical West AfricaCambridge University Press Cambridge Mass USA 1986
[23] K M Joppa A Vovor K Eklu-Gadegbeku A Agbonon KAklikokou and M Gbeassor ldquoEffect of Morinda Lucida benth(Rubiaceae) and Newbouldia leavis P beauv (Bignoniaceae) onsickling of red blood cellsrdquoMedecine Tropicale vol 68 no 3 pp251ndash256 2008
[24] K Kovendan K Murugan S P Shanthakumar S Vincentand J Hwang ldquoLarvicidal activity of Morinda citrifolia L(Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012
[25] Z Ilboudo L C B Dabire R C H Nebie et al ldquoBiologicalactivity and persistence of four essential oils towards themain pest of stored cowpeas Callosobruchus maculatus (F)(Coleoptera Bruchidae)rdquo Journal of Stored Products Researchvol 46 no 2 pp 124ndash128 2010
[26] R W D Taylor ldquoPhosphine a major fumigant at riskrdquo Interna-tional Pest Control vol 31 pp 10ndash14 1989
[27] W S Abbott ldquoA method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 pp 265ndash267 1925
[28] D J Finney Probit Analysis Cambridge University Press Cam-bridge UK 3rd edition 1971
[29] S A Bankole ldquoEffect of essential oils from two Nigerianmedicinal plants (Azadirachta indica and Morinda lucida)on growth and aflatoxin B1 production in maize grain by atoxigenicAspergillus flavusrdquoLetters inAppliedMicrobiology vol24 no 3 pp 190ndash192 1997
[30] J S Ashidi P J Houghton P J Hylands and T EfferthldquoEthnobotanical survey and cytotoxicity testing of plants ofSouth-western Nigeria used to treat cancer with isolationof cytotoxic constituents from Cajanus cajan Millsp leavesrdquoJournal of Ethnopharmacology vol 128 no 2 pp 501ndash512 2010
[31] L Lajide C O Adedire W A Muse and S O AgeleldquoInsecticidal activity of Powders of some Nigerian plant againstthe maize weevils (Sitophilus zeamais Mots) in stored maizerdquoEntomology and the Nigerian Economy vol 31 pp 227ndash2351998
[32] T S L Ngamo I Ngatanko M B Ngassoum P M Mapong-mestsem and T Hance ldquoPersistence of insecticidal activities ofcrude essential oils of three aromatic plants towards four majorstored product insect pestsrdquo African Journal of AgriculturalResearch vol 2 pp 173ndash177 2007
[33] M Negahban and S Moharramipour ldquoFumigant toxicityof Eucalyptus intertexta Eucalyptus sargentii and Eucalyp-tus camaldulensis against stored-product beetlesrdquo Journal ofApplied Entomology vol 131 no 4 pp 256ndash261 2007
[34] A F Ndomo L A Tapondjou L T Ngamo and T HanceldquoInsecticidal activities of essential oil of Callistemon viminalisapplied as fumigant and powder against two bruchidsrdquo Journalof Applied Entomology vol 134 no 4 pp 333ndash341 2010
[35] M S Owolabi M O Oladimeji L Lajide G Singh PMarimuthu and V A Isidorov ldquoBioactivity of three plantderived essential oils agianst the maize weevils Sitophilus zea-mais(Motschulsky) and cowpea weevils Callosobruchus macu-latus (Fabricius)rdquo Electronic Journal of Environmental Agricul-tural and Food Chemistry vol 8 no 9 pp 828ndash835 2009
[36] M K Chaubey ldquoFumigant toxicity of essential oils from somecommon spices against pulse beetle Callosobruchus chinensis(Coleoptera Bruchidae)rdquo Journal of Oleo Science vol 57 no 3pp 171ndash179 2008
[37] S Moharramipour A Taghizadeh M H Meshkatalsadat AA Talebi and Y Fathipour ldquoRepellent and fumigant toxicity ofessential oil fromThymus persicus against Tribolium castaneumand Callosobruchus maculatusrdquo Communications in Agricul-tural and Applied Biological Sciences vol 73 no 3 pp 639ndash6422008
[38] S O Okoh O T Asekun O B Familoni and A J AfolayanldquoComposition and antioxidant activities of leaf and root volatileoils ofMorinda lucidardquoNatural Product Communications vol 6no 10 pp 1537ndash1541 2011
[39] K Koumaglo M Gbeassor O Nikabu C de Souza and WWerner ldquoEffects of three compounds extracted from Morindalucida on Plasmodium falciparumrdquo Planta Medica vol 58 no6 pp 533ndash534 1992
[40] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis (Rondani) (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 no 4 pp 363ndash371 2005
[41] R Islam R I Khan S M Al-Reza Y T Jeong C H Song andM Khalequzzaman ldquoChemical composition and insecticidalproperties of Cinnamomum aromaticum (Nees) essential oilagainst the stored product beetleCallosobruchusmaculatus (F)rdquoJournal of the Science of Food and Agriculture vol 89 no 7 pp1241ndash1246 2009
[42] W S Nyamador G K Ketoh K Amevoin Y Nuto H KKoumaglo and I A Glitho ldquoVariation in the susceptibility oftwo Callosobruchus species to essential oilsrdquo Journal of StoredProducts Research vol 46 no 1 pp 48ndash51 2010
[43] H Kouninki T Hance F A Noudjou et al ldquoToxicity ofsome terpenoids of essential oils of Xylopia aethiopica fromCameroon against Sitophilus zeamais Motschulskyrdquo Journal ofApplied Entomology vol 131 no 4 pp 269ndash274 2007
[44] R Waliwitiya M B Isman R S Vernon and A RisemanldquoInsecticidal activity of selected monoterpenoids and rosemaryoil to Agriotes obscurus (Coleoptera Elateridae)rdquo Journal ofEconomic Entomology vol 98 no 5 pp 1560ndash1565 2005
[45] N S Iacobellis P Lo Cantore F Capasso and F SenatoreldquoAntibacterial activity of Cuminum cyminum L and Carumcarvi L essential oilsrdquo Journal of Agricultural and Food Chem-istry vol 53 no 1 pp 57ndash61 2005
Component KIa 120573-Caryophyllene 1421 Tr119901-Cymen-7-ol-acetate 1423 Tr(119864)-Cinnamyl acetate 1450 Tr(119864)-Methyl isoeugenol 1495 TrBicyclogermacrene 1497 Tr120572-Bulnesene 1506 Tr120573-Sesquiphellandrene 1526 TrElemol 1554 TrElemicin 1557 TrSpathulenol 1578 04Caryophyllene oxide 1583 05g-Eudesmol 1635 Tr120573-Eudesmol 1651 02120572-Eudesmol 1655 TrIntermedeol 1667 TrTotal identified 9540Major components are shown in boldlowastThese chemical components were identified by authentic compound injec-tionaKI Kovats retention indicesTr = trace amount lt01
467 and 867mortality after 24 h and 48 h at 020 120583gmLrespectively (Figures 1(a) and 1(b)) Both M lucida essen-tial oil and pesticides reached 100 mortality at 72 h at020120583gmL (Figure 1(c)) The negative control showed noactivity except at 72 h (17)
33 Determination of Insecticidal Activity Probit analysisshowed that M lucida essential oil after 24 h presentedan insecticidal activity (LC
90= 0629120583gmL) similar to
Phostoxin (LC90
= 0652120583gmL) and Primo-ban-20 (LC90
= 0726 120583gmL) At 24 h essential oil of M lucida (LC50
= 0298 120583gmL) and Phostoxin (LC50
= 0295 120583gmL) weremore efficient than Primo-ban-20 (LC
50= 0393 120583gmL) At
48 h after exposure there was no significant difference inLC50
between M lucida essential oil compared to Primo-ban-20 and Phostoxin (Table 2 where all confidence intervalsoverlap)
4 Discussion
Recently biological activity has been demonstrated for theM lucida essential oil such as the complete suppression ofaflatoxin synthesis in maize [29] cytotoxic activity on cancercell lines [30] and antioxidant and anti-inflammatory activity[20] This study demonstrated a potent insecticidal activityfor M lucida showing 100 mortality on C maculatus atconcentrations of 020 and 040120583gmL after 72 h (Figure 1)
Our results for M lucida essential oil are consistent withother reports of essential oils that exhibited insecticidal andrepellency activity against C maculatus such as Eucalyp-tus intertexta Callistemon viminalis Ocimum americanum
Hyptis suaveolens Hyptis spicigera Lippia multiflora Cym-bopogon citratus and Lippia rugosa [25 31ndash35] Chaubey[36] evaluated the insecticidal oviposition egg hatchingand developmental inhibitory activities of seven differentessential oils against Callosobruchus chinensis and found thatNigella sativa was the most effective at all stages MoreoverKeita et al [16] found that essential oils ofOcimum basilicumand O gratissimum at a dose of 25 120583gmL after 12 h offumigation produced 80 and 70 mortality respectivelySimilarly Moharramipour et al [37] have shown a repellencyof 824 of the Thymus persicus essential oil against Cmaculatus
The chemical components of this essential oil have beenpreviously reported [38 39] although there were differencesin the ratio of their chemicals constituents Okoh et al [38]identified 50 compounds where 120572-terpinene was the majorconstituent with 178 while in this study it represented only08 This difference could be attributed to the number ofcompounds identified in each study since in this study 63components were identified representing 954 of the totaloil content The essential oil was found to be dominated byoxygenated monoterpenes (Table 1)
Several essential oils components have shown insecti-cidal activity against C maculatus The leaf oil of Cymbo-pogon schoenanthus rich in piperitone gave 90 mortalityafter 24 h at a concentration of 67120583LL [40] Plectranthusgrandifolius essential oil predominantly (E)-myroxide wasshown to be toxic to both adults and eggs of C maculatusLikewise Cinnamomum aromaticum bark oil rich in cis-cinnamaldehyde was insecticidal with an LC
50of 276120583gcm2
after 24 h [41] The essential oil from Cymbopogon giganteusrich in limonene and p-mentha-1(7)8-dien-2-ol demon-strated insecticidal activity against C maculatus and Csubinnotatus [42]The concentrations of themajor terpenoidsconstituents presented here for M lucida essential oil areconsistent with use of this plant as an insecticide and insectrepellent
The insecticidal activity of the M lucida essential oilcould be attributed to those known major components ofoxygenated monoterpenes 18-cineole (434) 120572-terpinylacetate (145) 4-terpinen-4-ol (34) 120572-terpinol (34)and monoterpene hydrocarbons sabinene (82) and 120573-pinene (40) These are compounds whose insecticide andrepellent activities have been reported previously [32 43 44]Nevertheless it also has been shown that minor componentsmay contribute to the biological activity [45] such asMyrcene120572-phellandrene and Camphene
The intensive use of synthetic insecticides not only pro-duces genetic resistance by insect species but also causes seri-ous problems of toxicity to humans and other animal species[6 7] Therefore the essential oil extracted from M lucidarepresents a botanical insecticide source since it showed apotent insecticidal activity comparable to both pesticidesPhostoxin and Primo-ban-20 (Table 2 where confidenceintervals overlap) The lack of a statistical difference betweenM lucida essential oil and two commercial pesticides is apositive result and demonstrates their equivalence thoughwecannot conclude thatM lucida is more effective
The Scientific World Journal 5
0
10
20
30
40
50
60
70
005 01 02 04
Mortality at 24h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(a)
PhostoxinPrimo-ban-20
0102030405060708090
100
005 01 02 04
Mortality at 48h
(120583gmL)
M lucida
(b)
0102030405060708090
100
005 01 02 04
Mortality at 72h
PhostoxinPrimo-ban-20
(120583gmL)
M lucida
(c)
Figure 1 Effect of theMorinda lucida essential oil and pesticides against Callosobruchus maculatus at different concentrations and exposuretimes
Table 2 Insecticidal activity ofM lucida essential oil against C maculatus
Units LC50 (concentration causing 50 mortality) and LC90 (concentration causing 90 mortality) in units of 120583gmL 95 confidence interval (CI) is shownin parenthesis Insecticidal activity is considered significantly different when the 95 CI fail to overlap aChi-square value of Probit model fit all significant at119875 lt 005 level
6 The Scientific World Journal
The results obtained indicate that M lucida essential oilwas biocidal even at lower concentration and may thereforebe considered to be a useful alternative to synthetic insecti-cides
5 Conclusions
This study demonstrated that essential oil extracted fromMlucida was toxic to pulse beetle C maculatus Therefore itcould be used as an alternative strategy for C maculatuscontrol and as a substitute for synthetic pesticides Howeverfurther studies are necessary to elucidate the mode of actionand their environmental impact and develop formulations toimprove the insecticidal efficacy This would benefit agricul-tural sectors of developing countries as these essential oils arereadily available and biodegradableThe problem of volatilitycan be resolved through a controlled release formulation oftheir active chemical compositions
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Moses S Owolabi and Eduardo Padilla-Camberos con-tributed equally to this work
Acknowledgments
The authors thank Damilola Ajayi for the collection andidentification of the plant sample Mrs Helen Owolabi isgratefully acknowledged for her useful comments
References
[1] C Park S I Kim and Y J Ahn ldquoInsecticidal activity ofasarones identified in Acorus gramineus rhizome against threecoleopteran stored-product insectsrdquo Journal of Stored ProductsResearch vol 39 no 3 pp 333ndash342 2003
[2] A Rahman and F A Talukder ldquoBioefficacy of some plantderivatives that protect grain against the pulse beetle Calloso-bruchus maculatusrdquo Journal of Insect Science vol 6 no 3 pp1ndash10 2006
[3] B Mulatu and T Gebrmedhin ldquoOviposition-deterrent andtoxic effects of various botanicals on the Adzuki bean beetleCallosobruchus chinensis Lrdquo International Journal of TropicalInsect Science vol 20 pp 33ndash38 2000
[4] F A Ajayi and N E S Lale ldquoSeed coat texture host species andtime of application affect the efficacy of essential oils appliedfor the control of Callosobruchus maculatus (F) (ColeopteraBruchidae) in stored pulsesrdquo International Journal of PestManagement vol 47 no 3 pp 161ndash166 2001
[5] L A Tapondjou C Adler H Bouda and D A FontemldquoEfficacy of powder and essential oil fromChenopodium ambro-sioides leaves as post-harvest grain protectants against six-stored product beetlesrdquo Journal of Stored Products Research vol38 no 4 pp 395ndash402 2002
[6] M K Mahmud M M H Khan M Husain M I Alam andM S I Afrad ldquoToxic effects of different plant oils on pulsebeetle Callosobruchus chinensis Linn (Coleoptera Bruchidae)rdquoJournal of the Asiatic Society of Bangladesh Science vol 28 pp11ndash18 2002
[7] FA Lolestani andN Shayesteh ldquoFumigant toxicity ofZiziphoraclinopodioides (Boiss) (Lamiaceae) against adults and eggsof Callosobruchus maculatus (Fab) (Coleoptera Bruchidae)rdquoJournal of Biological Sciences vol 9 no 1 pp 92ndash95 2009
[8] E A Elhag ldquoDeterrent effects of some botanical products onoviposition of the cowpea bruchid Callosobruchus maculatus(F) (Coleoptera Bruchidae)rdquo International Journal of PestManagement vol 46 no 2 pp 109ndash113 2000
[9] C O Adedire and J O Akinneye ldquoBiological activity oftree marigold Tithonia diversifolia on cowpea seed bruchidCallosobruchus maculatus (Coleoptera Bruchidae)rdquo Annals ofApplied Biology vol 144 pp 185ndash189 2004
[10] P Koona and S Dorn ldquoExtracts from Tephrosia vogelii forthe protection of stored legume seeds against damage by threebruchid speciesrdquo Annals of Applied Biology vol 147 no 1 pp43ndash48 2005
[11] D P Papachristos A C Kimbaris N T Papadopoulos and MG Polissiou ldquoToxicity of citrus essential oils against Ceratitiscapitata (Diptera Tephritidae) larvaerdquo Annals of Applied Biol-ogy vol 155 no 3 pp 381ndash389 2009
[12] M Martinez-Velazquez G A Castillo-Herrera R Rosario-Cruz et al ldquoAcaricidal effect and chemical composition ofessential oils extracted from Cuminum cyminum Pimentadioica and Ocimum basilicum against the cattle tick Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo ParasitologyResearch vol 108 no 2 pp 481ndash487 2011
[13] R S Kawuki A Agona P Nampala and E Adipala ldquoA compar-ison of effectiveness of plant-based and synthetic insecticides inthe field management of pod and storage pests of cowpeardquoCropProtection vol 24 no 5 pp 473ndash478 2005
[14] I Mahfuz and M Khalequzzaman ldquoContact and fumiganttoxicity of essential oils against Callosobruchus maculatusrdquoUniversity Journal of Zoology Rajshahi University vol 26 pp63ndash66 2007
[15] I Chiffelle A Huerta M Celis and J E Araya ldquoProximalanalysis and insecticidal effects of extracts from pepper tree(Schinus molle) leaves on elm leaf beetle (Xanthogalerucaluteola) larvaerdquo Industrial Crops and Products vol 43 no 1 pp523ndash528 2013
[16] S M Keita C Vincent J Schmit J T Arnason and ABelanger ldquoEfficacy of essential oil of Ocimum basilicum Land O gratissimum L applied as an insecticidal fumigant andpowder to controlCallosobruchusmaculatus (Fab) [ColeopteraBruchidae]rdquo Journal of Stored Products Research vol 37 no 4pp 339ndash349 2001
[17] M J Pascual-Villalobos and M C Ballesta-Acosta ldquoChemicalvariation in an Ocimum basilicum germplasm collection andactivity of the essential oils on Callosobruchus maculatusrdquoBiochemical Systematics and Ecology vol 31 no 7 pp 673ndash6792003
[18] R Pavela ldquoInsecticidal and repellent activity of selected essen-tial oils against of the pollen beetle Meligethes aeneus (Fabri-cius) adultsrdquo Industrial Crops and Products vol 34 no 1 pp888ndash892 2011
[19] M Martinez-Velazquez R Rosario-Cruz G Castillo-HerreraJ M Flores-Fernandez A H Alvarez and E Lugo-Cervantes
The Scientific World Journal 7
ldquoAcaricidal effect of EssentialOils fromLippia graveolens (Lami-ales Verbenaceae) Rosmarinus officinalis (Lamiales Lami-aceae) and Allium sativum (Liliales Liliaceae) Against Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo Journal ofMedical Entomology vol 48 no 4 pp 822ndash827 2011
[20] S D Karou T Tchacondo D P Ilboudo and J Simpore ldquoSaha-ran Rubiaceae a review of their traditional uses phytochem-istry and biological activitiesrdquo Pakistan Journal of BiologicalSciences vol 14 no 3 pp 149ndash169 2011
[21] J MMakinde and P O Obih ldquoScreening ofMorinda lucida leafextract for antimalarial action on Plasmodium berghei in micerdquoAfrican Journal of Medicine andMedical Sciences vol 17 pp 59ndash63 1985
[22] B Oliver-Bever Medicinal Plants in Tropical West AfricaCambridge University Press Cambridge Mass USA 1986
[23] K M Joppa A Vovor K Eklu-Gadegbeku A Agbonon KAklikokou and M Gbeassor ldquoEffect of Morinda Lucida benth(Rubiaceae) and Newbouldia leavis P beauv (Bignoniaceae) onsickling of red blood cellsrdquoMedecine Tropicale vol 68 no 3 pp251ndash256 2008
[24] K Kovendan K Murugan S P Shanthakumar S Vincentand J Hwang ldquoLarvicidal activity of Morinda citrifolia L(Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012
[25] Z Ilboudo L C B Dabire R C H Nebie et al ldquoBiologicalactivity and persistence of four essential oils towards themain pest of stored cowpeas Callosobruchus maculatus (F)(Coleoptera Bruchidae)rdquo Journal of Stored Products Researchvol 46 no 2 pp 124ndash128 2010
[26] R W D Taylor ldquoPhosphine a major fumigant at riskrdquo Interna-tional Pest Control vol 31 pp 10ndash14 1989
[27] W S Abbott ldquoA method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 pp 265ndash267 1925
[28] D J Finney Probit Analysis Cambridge University Press Cam-bridge UK 3rd edition 1971
[29] S A Bankole ldquoEffect of essential oils from two Nigerianmedicinal plants (Azadirachta indica and Morinda lucida)on growth and aflatoxin B1 production in maize grain by atoxigenicAspergillus flavusrdquoLetters inAppliedMicrobiology vol24 no 3 pp 190ndash192 1997
[30] J S Ashidi P J Houghton P J Hylands and T EfferthldquoEthnobotanical survey and cytotoxicity testing of plants ofSouth-western Nigeria used to treat cancer with isolationof cytotoxic constituents from Cajanus cajan Millsp leavesrdquoJournal of Ethnopharmacology vol 128 no 2 pp 501ndash512 2010
[31] L Lajide C O Adedire W A Muse and S O AgeleldquoInsecticidal activity of Powders of some Nigerian plant againstthe maize weevils (Sitophilus zeamais Mots) in stored maizerdquoEntomology and the Nigerian Economy vol 31 pp 227ndash2351998
[32] T S L Ngamo I Ngatanko M B Ngassoum P M Mapong-mestsem and T Hance ldquoPersistence of insecticidal activities ofcrude essential oils of three aromatic plants towards four majorstored product insect pestsrdquo African Journal of AgriculturalResearch vol 2 pp 173ndash177 2007
[33] M Negahban and S Moharramipour ldquoFumigant toxicityof Eucalyptus intertexta Eucalyptus sargentii and Eucalyp-tus camaldulensis against stored-product beetlesrdquo Journal ofApplied Entomology vol 131 no 4 pp 256ndash261 2007
[34] A F Ndomo L A Tapondjou L T Ngamo and T HanceldquoInsecticidal activities of essential oil of Callistemon viminalisapplied as fumigant and powder against two bruchidsrdquo Journalof Applied Entomology vol 134 no 4 pp 333ndash341 2010
[35] M S Owolabi M O Oladimeji L Lajide G Singh PMarimuthu and V A Isidorov ldquoBioactivity of three plantderived essential oils agianst the maize weevils Sitophilus zea-mais(Motschulsky) and cowpea weevils Callosobruchus macu-latus (Fabricius)rdquo Electronic Journal of Environmental Agricul-tural and Food Chemistry vol 8 no 9 pp 828ndash835 2009
[36] M K Chaubey ldquoFumigant toxicity of essential oils from somecommon spices against pulse beetle Callosobruchus chinensis(Coleoptera Bruchidae)rdquo Journal of Oleo Science vol 57 no 3pp 171ndash179 2008
[37] S Moharramipour A Taghizadeh M H Meshkatalsadat AA Talebi and Y Fathipour ldquoRepellent and fumigant toxicity ofessential oil fromThymus persicus against Tribolium castaneumand Callosobruchus maculatusrdquo Communications in Agricul-tural and Applied Biological Sciences vol 73 no 3 pp 639ndash6422008
[38] S O Okoh O T Asekun O B Familoni and A J AfolayanldquoComposition and antioxidant activities of leaf and root volatileoils ofMorinda lucidardquoNatural Product Communications vol 6no 10 pp 1537ndash1541 2011
[39] K Koumaglo M Gbeassor O Nikabu C de Souza and WWerner ldquoEffects of three compounds extracted from Morindalucida on Plasmodium falciparumrdquo Planta Medica vol 58 no6 pp 533ndash534 1992
[40] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis (Rondani) (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 no 4 pp 363ndash371 2005
[41] R Islam R I Khan S M Al-Reza Y T Jeong C H Song andM Khalequzzaman ldquoChemical composition and insecticidalproperties of Cinnamomum aromaticum (Nees) essential oilagainst the stored product beetleCallosobruchusmaculatus (F)rdquoJournal of the Science of Food and Agriculture vol 89 no 7 pp1241ndash1246 2009
[42] W S Nyamador G K Ketoh K Amevoin Y Nuto H KKoumaglo and I A Glitho ldquoVariation in the susceptibility oftwo Callosobruchus species to essential oilsrdquo Journal of StoredProducts Research vol 46 no 1 pp 48ndash51 2010
[43] H Kouninki T Hance F A Noudjou et al ldquoToxicity ofsome terpenoids of essential oils of Xylopia aethiopica fromCameroon against Sitophilus zeamais Motschulskyrdquo Journal ofApplied Entomology vol 131 no 4 pp 269ndash274 2007
[44] R Waliwitiya M B Isman R S Vernon and A RisemanldquoInsecticidal activity of selected monoterpenoids and rosemaryoil to Agriotes obscurus (Coleoptera Elateridae)rdquo Journal ofEconomic Entomology vol 98 no 5 pp 1560ndash1565 2005
[45] N S Iacobellis P Lo Cantore F Capasso and F SenatoreldquoAntibacterial activity of Cuminum cyminum L and Carumcarvi L essential oilsrdquo Journal of Agricultural and Food Chem-istry vol 53 no 1 pp 57ndash61 2005
Units LC50 (concentration causing 50 mortality) and LC90 (concentration causing 90 mortality) in units of 120583gmL 95 confidence interval (CI) is shownin parenthesis Insecticidal activity is considered significantly different when the 95 CI fail to overlap aChi-square value of Probit model fit all significant at119875 lt 005 level
6 The Scientific World Journal
The results obtained indicate that M lucida essential oilwas biocidal even at lower concentration and may thereforebe considered to be a useful alternative to synthetic insecti-cides
5 Conclusions
This study demonstrated that essential oil extracted fromMlucida was toxic to pulse beetle C maculatus Therefore itcould be used as an alternative strategy for C maculatuscontrol and as a substitute for synthetic pesticides Howeverfurther studies are necessary to elucidate the mode of actionand their environmental impact and develop formulations toimprove the insecticidal efficacy This would benefit agricul-tural sectors of developing countries as these essential oils arereadily available and biodegradableThe problem of volatilitycan be resolved through a controlled release formulation oftheir active chemical compositions
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Moses S Owolabi and Eduardo Padilla-Camberos con-tributed equally to this work
Acknowledgments
The authors thank Damilola Ajayi for the collection andidentification of the plant sample Mrs Helen Owolabi isgratefully acknowledged for her useful comments
References
[1] C Park S I Kim and Y J Ahn ldquoInsecticidal activity ofasarones identified in Acorus gramineus rhizome against threecoleopteran stored-product insectsrdquo Journal of Stored ProductsResearch vol 39 no 3 pp 333ndash342 2003
[2] A Rahman and F A Talukder ldquoBioefficacy of some plantderivatives that protect grain against the pulse beetle Calloso-bruchus maculatusrdquo Journal of Insect Science vol 6 no 3 pp1ndash10 2006
[3] B Mulatu and T Gebrmedhin ldquoOviposition-deterrent andtoxic effects of various botanicals on the Adzuki bean beetleCallosobruchus chinensis Lrdquo International Journal of TropicalInsect Science vol 20 pp 33ndash38 2000
[4] F A Ajayi and N E S Lale ldquoSeed coat texture host species andtime of application affect the efficacy of essential oils appliedfor the control of Callosobruchus maculatus (F) (ColeopteraBruchidae) in stored pulsesrdquo International Journal of PestManagement vol 47 no 3 pp 161ndash166 2001
[5] L A Tapondjou C Adler H Bouda and D A FontemldquoEfficacy of powder and essential oil fromChenopodium ambro-sioides leaves as post-harvest grain protectants against six-stored product beetlesrdquo Journal of Stored Products Research vol38 no 4 pp 395ndash402 2002
[6] M K Mahmud M M H Khan M Husain M I Alam andM S I Afrad ldquoToxic effects of different plant oils on pulsebeetle Callosobruchus chinensis Linn (Coleoptera Bruchidae)rdquoJournal of the Asiatic Society of Bangladesh Science vol 28 pp11ndash18 2002
[7] FA Lolestani andN Shayesteh ldquoFumigant toxicity ofZiziphoraclinopodioides (Boiss) (Lamiaceae) against adults and eggsof Callosobruchus maculatus (Fab) (Coleoptera Bruchidae)rdquoJournal of Biological Sciences vol 9 no 1 pp 92ndash95 2009
[8] E A Elhag ldquoDeterrent effects of some botanical products onoviposition of the cowpea bruchid Callosobruchus maculatus(F) (Coleoptera Bruchidae)rdquo International Journal of PestManagement vol 46 no 2 pp 109ndash113 2000
[9] C O Adedire and J O Akinneye ldquoBiological activity oftree marigold Tithonia diversifolia on cowpea seed bruchidCallosobruchus maculatus (Coleoptera Bruchidae)rdquo Annals ofApplied Biology vol 144 pp 185ndash189 2004
[10] P Koona and S Dorn ldquoExtracts from Tephrosia vogelii forthe protection of stored legume seeds against damage by threebruchid speciesrdquo Annals of Applied Biology vol 147 no 1 pp43ndash48 2005
[11] D P Papachristos A C Kimbaris N T Papadopoulos and MG Polissiou ldquoToxicity of citrus essential oils against Ceratitiscapitata (Diptera Tephritidae) larvaerdquo Annals of Applied Biol-ogy vol 155 no 3 pp 381ndash389 2009
[12] M Martinez-Velazquez G A Castillo-Herrera R Rosario-Cruz et al ldquoAcaricidal effect and chemical composition ofessential oils extracted from Cuminum cyminum Pimentadioica and Ocimum basilicum against the cattle tick Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo ParasitologyResearch vol 108 no 2 pp 481ndash487 2011
[13] R S Kawuki A Agona P Nampala and E Adipala ldquoA compar-ison of effectiveness of plant-based and synthetic insecticides inthe field management of pod and storage pests of cowpeardquoCropProtection vol 24 no 5 pp 473ndash478 2005
[14] I Mahfuz and M Khalequzzaman ldquoContact and fumiganttoxicity of essential oils against Callosobruchus maculatusrdquoUniversity Journal of Zoology Rajshahi University vol 26 pp63ndash66 2007
[15] I Chiffelle A Huerta M Celis and J E Araya ldquoProximalanalysis and insecticidal effects of extracts from pepper tree(Schinus molle) leaves on elm leaf beetle (Xanthogalerucaluteola) larvaerdquo Industrial Crops and Products vol 43 no 1 pp523ndash528 2013
[16] S M Keita C Vincent J Schmit J T Arnason and ABelanger ldquoEfficacy of essential oil of Ocimum basilicum Land O gratissimum L applied as an insecticidal fumigant andpowder to controlCallosobruchusmaculatus (Fab) [ColeopteraBruchidae]rdquo Journal of Stored Products Research vol 37 no 4pp 339ndash349 2001
[17] M J Pascual-Villalobos and M C Ballesta-Acosta ldquoChemicalvariation in an Ocimum basilicum germplasm collection andactivity of the essential oils on Callosobruchus maculatusrdquoBiochemical Systematics and Ecology vol 31 no 7 pp 673ndash6792003
[18] R Pavela ldquoInsecticidal and repellent activity of selected essen-tial oils against of the pollen beetle Meligethes aeneus (Fabri-cius) adultsrdquo Industrial Crops and Products vol 34 no 1 pp888ndash892 2011
[19] M Martinez-Velazquez R Rosario-Cruz G Castillo-HerreraJ M Flores-Fernandez A H Alvarez and E Lugo-Cervantes
The Scientific World Journal 7
ldquoAcaricidal effect of EssentialOils fromLippia graveolens (Lami-ales Verbenaceae) Rosmarinus officinalis (Lamiales Lami-aceae) and Allium sativum (Liliales Liliaceae) Against Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo Journal ofMedical Entomology vol 48 no 4 pp 822ndash827 2011
[20] S D Karou T Tchacondo D P Ilboudo and J Simpore ldquoSaha-ran Rubiaceae a review of their traditional uses phytochem-istry and biological activitiesrdquo Pakistan Journal of BiologicalSciences vol 14 no 3 pp 149ndash169 2011
[21] J MMakinde and P O Obih ldquoScreening ofMorinda lucida leafextract for antimalarial action on Plasmodium berghei in micerdquoAfrican Journal of Medicine andMedical Sciences vol 17 pp 59ndash63 1985
[22] B Oliver-Bever Medicinal Plants in Tropical West AfricaCambridge University Press Cambridge Mass USA 1986
[23] K M Joppa A Vovor K Eklu-Gadegbeku A Agbonon KAklikokou and M Gbeassor ldquoEffect of Morinda Lucida benth(Rubiaceae) and Newbouldia leavis P beauv (Bignoniaceae) onsickling of red blood cellsrdquoMedecine Tropicale vol 68 no 3 pp251ndash256 2008
[24] K Kovendan K Murugan S P Shanthakumar S Vincentand J Hwang ldquoLarvicidal activity of Morinda citrifolia L(Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012
[25] Z Ilboudo L C B Dabire R C H Nebie et al ldquoBiologicalactivity and persistence of four essential oils towards themain pest of stored cowpeas Callosobruchus maculatus (F)(Coleoptera Bruchidae)rdquo Journal of Stored Products Researchvol 46 no 2 pp 124ndash128 2010
[26] R W D Taylor ldquoPhosphine a major fumigant at riskrdquo Interna-tional Pest Control vol 31 pp 10ndash14 1989
[27] W S Abbott ldquoA method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 pp 265ndash267 1925
[28] D J Finney Probit Analysis Cambridge University Press Cam-bridge UK 3rd edition 1971
[29] S A Bankole ldquoEffect of essential oils from two Nigerianmedicinal plants (Azadirachta indica and Morinda lucida)on growth and aflatoxin B1 production in maize grain by atoxigenicAspergillus flavusrdquoLetters inAppliedMicrobiology vol24 no 3 pp 190ndash192 1997
[30] J S Ashidi P J Houghton P J Hylands and T EfferthldquoEthnobotanical survey and cytotoxicity testing of plants ofSouth-western Nigeria used to treat cancer with isolationof cytotoxic constituents from Cajanus cajan Millsp leavesrdquoJournal of Ethnopharmacology vol 128 no 2 pp 501ndash512 2010
[31] L Lajide C O Adedire W A Muse and S O AgeleldquoInsecticidal activity of Powders of some Nigerian plant againstthe maize weevils (Sitophilus zeamais Mots) in stored maizerdquoEntomology and the Nigerian Economy vol 31 pp 227ndash2351998
[32] T S L Ngamo I Ngatanko M B Ngassoum P M Mapong-mestsem and T Hance ldquoPersistence of insecticidal activities ofcrude essential oils of three aromatic plants towards four majorstored product insect pestsrdquo African Journal of AgriculturalResearch vol 2 pp 173ndash177 2007
[33] M Negahban and S Moharramipour ldquoFumigant toxicityof Eucalyptus intertexta Eucalyptus sargentii and Eucalyp-tus camaldulensis against stored-product beetlesrdquo Journal ofApplied Entomology vol 131 no 4 pp 256ndash261 2007
[34] A F Ndomo L A Tapondjou L T Ngamo and T HanceldquoInsecticidal activities of essential oil of Callistemon viminalisapplied as fumigant and powder against two bruchidsrdquo Journalof Applied Entomology vol 134 no 4 pp 333ndash341 2010
[35] M S Owolabi M O Oladimeji L Lajide G Singh PMarimuthu and V A Isidorov ldquoBioactivity of three plantderived essential oils agianst the maize weevils Sitophilus zea-mais(Motschulsky) and cowpea weevils Callosobruchus macu-latus (Fabricius)rdquo Electronic Journal of Environmental Agricul-tural and Food Chemistry vol 8 no 9 pp 828ndash835 2009
[36] M K Chaubey ldquoFumigant toxicity of essential oils from somecommon spices against pulse beetle Callosobruchus chinensis(Coleoptera Bruchidae)rdquo Journal of Oleo Science vol 57 no 3pp 171ndash179 2008
[37] S Moharramipour A Taghizadeh M H Meshkatalsadat AA Talebi and Y Fathipour ldquoRepellent and fumigant toxicity ofessential oil fromThymus persicus against Tribolium castaneumand Callosobruchus maculatusrdquo Communications in Agricul-tural and Applied Biological Sciences vol 73 no 3 pp 639ndash6422008
[38] S O Okoh O T Asekun O B Familoni and A J AfolayanldquoComposition and antioxidant activities of leaf and root volatileoils ofMorinda lucidardquoNatural Product Communications vol 6no 10 pp 1537ndash1541 2011
[39] K Koumaglo M Gbeassor O Nikabu C de Souza and WWerner ldquoEffects of three compounds extracted from Morindalucida on Plasmodium falciparumrdquo Planta Medica vol 58 no6 pp 533ndash534 1992
[40] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis (Rondani) (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 no 4 pp 363ndash371 2005
[41] R Islam R I Khan S M Al-Reza Y T Jeong C H Song andM Khalequzzaman ldquoChemical composition and insecticidalproperties of Cinnamomum aromaticum (Nees) essential oilagainst the stored product beetleCallosobruchusmaculatus (F)rdquoJournal of the Science of Food and Agriculture vol 89 no 7 pp1241ndash1246 2009
[42] W S Nyamador G K Ketoh K Amevoin Y Nuto H KKoumaglo and I A Glitho ldquoVariation in the susceptibility oftwo Callosobruchus species to essential oilsrdquo Journal of StoredProducts Research vol 46 no 1 pp 48ndash51 2010
[43] H Kouninki T Hance F A Noudjou et al ldquoToxicity ofsome terpenoids of essential oils of Xylopia aethiopica fromCameroon against Sitophilus zeamais Motschulskyrdquo Journal ofApplied Entomology vol 131 no 4 pp 269ndash274 2007
[44] R Waliwitiya M B Isman R S Vernon and A RisemanldquoInsecticidal activity of selected monoterpenoids and rosemaryoil to Agriotes obscurus (Coleoptera Elateridae)rdquo Journal ofEconomic Entomology vol 98 no 5 pp 1560ndash1565 2005
[45] N S Iacobellis P Lo Cantore F Capasso and F SenatoreldquoAntibacterial activity of Cuminum cyminum L and Carumcarvi L essential oilsrdquo Journal of Agricultural and Food Chem-istry vol 53 no 1 pp 57ndash61 2005
The results obtained indicate that M lucida essential oilwas biocidal even at lower concentration and may thereforebe considered to be a useful alternative to synthetic insecti-cides
5 Conclusions
This study demonstrated that essential oil extracted fromMlucida was toxic to pulse beetle C maculatus Therefore itcould be used as an alternative strategy for C maculatuscontrol and as a substitute for synthetic pesticides Howeverfurther studies are necessary to elucidate the mode of actionand their environmental impact and develop formulations toimprove the insecticidal efficacy This would benefit agricul-tural sectors of developing countries as these essential oils arereadily available and biodegradableThe problem of volatilitycan be resolved through a controlled release formulation oftheir active chemical compositions
Conflict of Interests
The authors declare that there is no conflict of interestsregarding the publication of this paper
Authorsrsquo Contribution
Moses S Owolabi and Eduardo Padilla-Camberos con-tributed equally to this work
Acknowledgments
The authors thank Damilola Ajayi for the collection andidentification of the plant sample Mrs Helen Owolabi isgratefully acknowledged for her useful comments
References
[1] C Park S I Kim and Y J Ahn ldquoInsecticidal activity ofasarones identified in Acorus gramineus rhizome against threecoleopteran stored-product insectsrdquo Journal of Stored ProductsResearch vol 39 no 3 pp 333ndash342 2003
[2] A Rahman and F A Talukder ldquoBioefficacy of some plantderivatives that protect grain against the pulse beetle Calloso-bruchus maculatusrdquo Journal of Insect Science vol 6 no 3 pp1ndash10 2006
[3] B Mulatu and T Gebrmedhin ldquoOviposition-deterrent andtoxic effects of various botanicals on the Adzuki bean beetleCallosobruchus chinensis Lrdquo International Journal of TropicalInsect Science vol 20 pp 33ndash38 2000
[4] F A Ajayi and N E S Lale ldquoSeed coat texture host species andtime of application affect the efficacy of essential oils appliedfor the control of Callosobruchus maculatus (F) (ColeopteraBruchidae) in stored pulsesrdquo International Journal of PestManagement vol 47 no 3 pp 161ndash166 2001
[5] L A Tapondjou C Adler H Bouda and D A FontemldquoEfficacy of powder and essential oil fromChenopodium ambro-sioides leaves as post-harvest grain protectants against six-stored product beetlesrdquo Journal of Stored Products Research vol38 no 4 pp 395ndash402 2002
[6] M K Mahmud M M H Khan M Husain M I Alam andM S I Afrad ldquoToxic effects of different plant oils on pulsebeetle Callosobruchus chinensis Linn (Coleoptera Bruchidae)rdquoJournal of the Asiatic Society of Bangladesh Science vol 28 pp11ndash18 2002
[7] FA Lolestani andN Shayesteh ldquoFumigant toxicity ofZiziphoraclinopodioides (Boiss) (Lamiaceae) against adults and eggsof Callosobruchus maculatus (Fab) (Coleoptera Bruchidae)rdquoJournal of Biological Sciences vol 9 no 1 pp 92ndash95 2009
[8] E A Elhag ldquoDeterrent effects of some botanical products onoviposition of the cowpea bruchid Callosobruchus maculatus(F) (Coleoptera Bruchidae)rdquo International Journal of PestManagement vol 46 no 2 pp 109ndash113 2000
[9] C O Adedire and J O Akinneye ldquoBiological activity oftree marigold Tithonia diversifolia on cowpea seed bruchidCallosobruchus maculatus (Coleoptera Bruchidae)rdquo Annals ofApplied Biology vol 144 pp 185ndash189 2004
[10] P Koona and S Dorn ldquoExtracts from Tephrosia vogelii forthe protection of stored legume seeds against damage by threebruchid speciesrdquo Annals of Applied Biology vol 147 no 1 pp43ndash48 2005
[11] D P Papachristos A C Kimbaris N T Papadopoulos and MG Polissiou ldquoToxicity of citrus essential oils against Ceratitiscapitata (Diptera Tephritidae) larvaerdquo Annals of Applied Biol-ogy vol 155 no 3 pp 381ndash389 2009
[12] M Martinez-Velazquez G A Castillo-Herrera R Rosario-Cruz et al ldquoAcaricidal effect and chemical composition ofessential oils extracted from Cuminum cyminum Pimentadioica and Ocimum basilicum against the cattle tick Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo ParasitologyResearch vol 108 no 2 pp 481ndash487 2011
[13] R S Kawuki A Agona P Nampala and E Adipala ldquoA compar-ison of effectiveness of plant-based and synthetic insecticides inthe field management of pod and storage pests of cowpeardquoCropProtection vol 24 no 5 pp 473ndash478 2005
[14] I Mahfuz and M Khalequzzaman ldquoContact and fumiganttoxicity of essential oils against Callosobruchus maculatusrdquoUniversity Journal of Zoology Rajshahi University vol 26 pp63ndash66 2007
[15] I Chiffelle A Huerta M Celis and J E Araya ldquoProximalanalysis and insecticidal effects of extracts from pepper tree(Schinus molle) leaves on elm leaf beetle (Xanthogalerucaluteola) larvaerdquo Industrial Crops and Products vol 43 no 1 pp523ndash528 2013
[16] S M Keita C Vincent J Schmit J T Arnason and ABelanger ldquoEfficacy of essential oil of Ocimum basilicum Land O gratissimum L applied as an insecticidal fumigant andpowder to controlCallosobruchusmaculatus (Fab) [ColeopteraBruchidae]rdquo Journal of Stored Products Research vol 37 no 4pp 339ndash349 2001
[17] M J Pascual-Villalobos and M C Ballesta-Acosta ldquoChemicalvariation in an Ocimum basilicum germplasm collection andactivity of the essential oils on Callosobruchus maculatusrdquoBiochemical Systematics and Ecology vol 31 no 7 pp 673ndash6792003
[18] R Pavela ldquoInsecticidal and repellent activity of selected essen-tial oils against of the pollen beetle Meligethes aeneus (Fabri-cius) adultsrdquo Industrial Crops and Products vol 34 no 1 pp888ndash892 2011
[19] M Martinez-Velazquez R Rosario-Cruz G Castillo-HerreraJ M Flores-Fernandez A H Alvarez and E Lugo-Cervantes
The Scientific World Journal 7
ldquoAcaricidal effect of EssentialOils fromLippia graveolens (Lami-ales Verbenaceae) Rosmarinus officinalis (Lamiales Lami-aceae) and Allium sativum (Liliales Liliaceae) Against Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo Journal ofMedical Entomology vol 48 no 4 pp 822ndash827 2011
[20] S D Karou T Tchacondo D P Ilboudo and J Simpore ldquoSaha-ran Rubiaceae a review of their traditional uses phytochem-istry and biological activitiesrdquo Pakistan Journal of BiologicalSciences vol 14 no 3 pp 149ndash169 2011
[21] J MMakinde and P O Obih ldquoScreening ofMorinda lucida leafextract for antimalarial action on Plasmodium berghei in micerdquoAfrican Journal of Medicine andMedical Sciences vol 17 pp 59ndash63 1985
[22] B Oliver-Bever Medicinal Plants in Tropical West AfricaCambridge University Press Cambridge Mass USA 1986
[23] K M Joppa A Vovor K Eklu-Gadegbeku A Agbonon KAklikokou and M Gbeassor ldquoEffect of Morinda Lucida benth(Rubiaceae) and Newbouldia leavis P beauv (Bignoniaceae) onsickling of red blood cellsrdquoMedecine Tropicale vol 68 no 3 pp251ndash256 2008
[24] K Kovendan K Murugan S P Shanthakumar S Vincentand J Hwang ldquoLarvicidal activity of Morinda citrifolia L(Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012
[25] Z Ilboudo L C B Dabire R C H Nebie et al ldquoBiologicalactivity and persistence of four essential oils towards themain pest of stored cowpeas Callosobruchus maculatus (F)(Coleoptera Bruchidae)rdquo Journal of Stored Products Researchvol 46 no 2 pp 124ndash128 2010
[26] R W D Taylor ldquoPhosphine a major fumigant at riskrdquo Interna-tional Pest Control vol 31 pp 10ndash14 1989
[27] W S Abbott ldquoA method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 pp 265ndash267 1925
[28] D J Finney Probit Analysis Cambridge University Press Cam-bridge UK 3rd edition 1971
[29] S A Bankole ldquoEffect of essential oils from two Nigerianmedicinal plants (Azadirachta indica and Morinda lucida)on growth and aflatoxin B1 production in maize grain by atoxigenicAspergillus flavusrdquoLetters inAppliedMicrobiology vol24 no 3 pp 190ndash192 1997
[30] J S Ashidi P J Houghton P J Hylands and T EfferthldquoEthnobotanical survey and cytotoxicity testing of plants ofSouth-western Nigeria used to treat cancer with isolationof cytotoxic constituents from Cajanus cajan Millsp leavesrdquoJournal of Ethnopharmacology vol 128 no 2 pp 501ndash512 2010
[31] L Lajide C O Adedire W A Muse and S O AgeleldquoInsecticidal activity of Powders of some Nigerian plant againstthe maize weevils (Sitophilus zeamais Mots) in stored maizerdquoEntomology and the Nigerian Economy vol 31 pp 227ndash2351998
[32] T S L Ngamo I Ngatanko M B Ngassoum P M Mapong-mestsem and T Hance ldquoPersistence of insecticidal activities ofcrude essential oils of three aromatic plants towards four majorstored product insect pestsrdquo African Journal of AgriculturalResearch vol 2 pp 173ndash177 2007
[33] M Negahban and S Moharramipour ldquoFumigant toxicityof Eucalyptus intertexta Eucalyptus sargentii and Eucalyp-tus camaldulensis against stored-product beetlesrdquo Journal ofApplied Entomology vol 131 no 4 pp 256ndash261 2007
[34] A F Ndomo L A Tapondjou L T Ngamo and T HanceldquoInsecticidal activities of essential oil of Callistemon viminalisapplied as fumigant and powder against two bruchidsrdquo Journalof Applied Entomology vol 134 no 4 pp 333ndash341 2010
[35] M S Owolabi M O Oladimeji L Lajide G Singh PMarimuthu and V A Isidorov ldquoBioactivity of three plantderived essential oils agianst the maize weevils Sitophilus zea-mais(Motschulsky) and cowpea weevils Callosobruchus macu-latus (Fabricius)rdquo Electronic Journal of Environmental Agricul-tural and Food Chemistry vol 8 no 9 pp 828ndash835 2009
[36] M K Chaubey ldquoFumigant toxicity of essential oils from somecommon spices against pulse beetle Callosobruchus chinensis(Coleoptera Bruchidae)rdquo Journal of Oleo Science vol 57 no 3pp 171ndash179 2008
[37] S Moharramipour A Taghizadeh M H Meshkatalsadat AA Talebi and Y Fathipour ldquoRepellent and fumigant toxicity ofessential oil fromThymus persicus against Tribolium castaneumand Callosobruchus maculatusrdquo Communications in Agricul-tural and Applied Biological Sciences vol 73 no 3 pp 639ndash6422008
[38] S O Okoh O T Asekun O B Familoni and A J AfolayanldquoComposition and antioxidant activities of leaf and root volatileoils ofMorinda lucidardquoNatural Product Communications vol 6no 10 pp 1537ndash1541 2011
[39] K Koumaglo M Gbeassor O Nikabu C de Souza and WWerner ldquoEffects of three compounds extracted from Morindalucida on Plasmodium falciparumrdquo Planta Medica vol 58 no6 pp 533ndash534 1992
[40] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis (Rondani) (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 no 4 pp 363ndash371 2005
[41] R Islam R I Khan S M Al-Reza Y T Jeong C H Song andM Khalequzzaman ldquoChemical composition and insecticidalproperties of Cinnamomum aromaticum (Nees) essential oilagainst the stored product beetleCallosobruchusmaculatus (F)rdquoJournal of the Science of Food and Agriculture vol 89 no 7 pp1241ndash1246 2009
[42] W S Nyamador G K Ketoh K Amevoin Y Nuto H KKoumaglo and I A Glitho ldquoVariation in the susceptibility oftwo Callosobruchus species to essential oilsrdquo Journal of StoredProducts Research vol 46 no 1 pp 48ndash51 2010
[43] H Kouninki T Hance F A Noudjou et al ldquoToxicity ofsome terpenoids of essential oils of Xylopia aethiopica fromCameroon against Sitophilus zeamais Motschulskyrdquo Journal ofApplied Entomology vol 131 no 4 pp 269ndash274 2007
[44] R Waliwitiya M B Isman R S Vernon and A RisemanldquoInsecticidal activity of selected monoterpenoids and rosemaryoil to Agriotes obscurus (Coleoptera Elateridae)rdquo Journal ofEconomic Entomology vol 98 no 5 pp 1560ndash1565 2005
[45] N S Iacobellis P Lo Cantore F Capasso and F SenatoreldquoAntibacterial activity of Cuminum cyminum L and Carumcarvi L essential oilsrdquo Journal of Agricultural and Food Chem-istry vol 53 no 1 pp 57ndash61 2005
ldquoAcaricidal effect of EssentialOils fromLippia graveolens (Lami-ales Verbenaceae) Rosmarinus officinalis (Lamiales Lami-aceae) and Allium sativum (Liliales Liliaceae) Against Rhipi-cephalus (Boophilus) microplus (Acari Ixodidae)rdquo Journal ofMedical Entomology vol 48 no 4 pp 822ndash827 2011
[20] S D Karou T Tchacondo D P Ilboudo and J Simpore ldquoSaha-ran Rubiaceae a review of their traditional uses phytochem-istry and biological activitiesrdquo Pakistan Journal of BiologicalSciences vol 14 no 3 pp 149ndash169 2011
[21] J MMakinde and P O Obih ldquoScreening ofMorinda lucida leafextract for antimalarial action on Plasmodium berghei in micerdquoAfrican Journal of Medicine andMedical Sciences vol 17 pp 59ndash63 1985
[22] B Oliver-Bever Medicinal Plants in Tropical West AfricaCambridge University Press Cambridge Mass USA 1986
[23] K M Joppa A Vovor K Eklu-Gadegbeku A Agbonon KAklikokou and M Gbeassor ldquoEffect of Morinda Lucida benth(Rubiaceae) and Newbouldia leavis P beauv (Bignoniaceae) onsickling of red blood cellsrdquoMedecine Tropicale vol 68 no 3 pp251ndash256 2008
[24] K Kovendan K Murugan S P Shanthakumar S Vincentand J Hwang ldquoLarvicidal activity of Morinda citrifolia L(Noni) (Family Rubiaceae) leaf extract against Anophelesstephensi Culex quinquefasciatus and Aedes aegyptirdquo Parasitol-ogy Research vol 111 no 4 pp 1481ndash1490 2012
[25] Z Ilboudo L C B Dabire R C H Nebie et al ldquoBiologicalactivity and persistence of four essential oils towards themain pest of stored cowpeas Callosobruchus maculatus (F)(Coleoptera Bruchidae)rdquo Journal of Stored Products Researchvol 46 no 2 pp 124ndash128 2010
[26] R W D Taylor ldquoPhosphine a major fumigant at riskrdquo Interna-tional Pest Control vol 31 pp 10ndash14 1989
[27] W S Abbott ldquoA method of computing the effectiveness of aninsecticiderdquo Journal of Economic Entomology vol 18 pp 265ndash267 1925
[28] D J Finney Probit Analysis Cambridge University Press Cam-bridge UK 3rd edition 1971
[29] S A Bankole ldquoEffect of essential oils from two Nigerianmedicinal plants (Azadirachta indica and Morinda lucida)on growth and aflatoxin B1 production in maize grain by atoxigenicAspergillus flavusrdquoLetters inAppliedMicrobiology vol24 no 3 pp 190ndash192 1997
[30] J S Ashidi P J Houghton P J Hylands and T EfferthldquoEthnobotanical survey and cytotoxicity testing of plants ofSouth-western Nigeria used to treat cancer with isolationof cytotoxic constituents from Cajanus cajan Millsp leavesrdquoJournal of Ethnopharmacology vol 128 no 2 pp 501ndash512 2010
[31] L Lajide C O Adedire W A Muse and S O AgeleldquoInsecticidal activity of Powders of some Nigerian plant againstthe maize weevils (Sitophilus zeamais Mots) in stored maizerdquoEntomology and the Nigerian Economy vol 31 pp 227ndash2351998
[32] T S L Ngamo I Ngatanko M B Ngassoum P M Mapong-mestsem and T Hance ldquoPersistence of insecticidal activities ofcrude essential oils of three aromatic plants towards four majorstored product insect pestsrdquo African Journal of AgriculturalResearch vol 2 pp 173ndash177 2007
[33] M Negahban and S Moharramipour ldquoFumigant toxicityof Eucalyptus intertexta Eucalyptus sargentii and Eucalyp-tus camaldulensis against stored-product beetlesrdquo Journal ofApplied Entomology vol 131 no 4 pp 256ndash261 2007
[34] A F Ndomo L A Tapondjou L T Ngamo and T HanceldquoInsecticidal activities of essential oil of Callistemon viminalisapplied as fumigant and powder against two bruchidsrdquo Journalof Applied Entomology vol 134 no 4 pp 333ndash341 2010
[35] M S Owolabi M O Oladimeji L Lajide G Singh PMarimuthu and V A Isidorov ldquoBioactivity of three plantderived essential oils agianst the maize weevils Sitophilus zea-mais(Motschulsky) and cowpea weevils Callosobruchus macu-latus (Fabricius)rdquo Electronic Journal of Environmental Agricul-tural and Food Chemistry vol 8 no 9 pp 828ndash835 2009
[36] M K Chaubey ldquoFumigant toxicity of essential oils from somecommon spices against pulse beetle Callosobruchus chinensis(Coleoptera Bruchidae)rdquo Journal of Oleo Science vol 57 no 3pp 171ndash179 2008
[37] S Moharramipour A Taghizadeh M H Meshkatalsadat AA Talebi and Y Fathipour ldquoRepellent and fumigant toxicity ofessential oil fromThymus persicus against Tribolium castaneumand Callosobruchus maculatusrdquo Communications in Agricul-tural and Applied Biological Sciences vol 73 no 3 pp 639ndash6422008
[38] S O Okoh O T Asekun O B Familoni and A J AfolayanldquoComposition and antioxidant activities of leaf and root volatileoils ofMorinda lucidardquoNatural Product Communications vol 6no 10 pp 1537ndash1541 2011
[39] K Koumaglo M Gbeassor O Nikabu C de Souza and WWerner ldquoEffects of three compounds extracted from Morindalucida on Plasmodium falciparumrdquo Planta Medica vol 58 no6 pp 533ndash534 1992
[40] G K Ketoh H K Koumaglo and I A Glitho ldquoInhibi-tion of Callosobruchus maculatus (F) (Coleoptera Bruchidae)development with essential oil extracted from Cymbopogonschoenanthus L Spreng (Poaceae) and the wasp Dinarmusbasalis (Rondani) (Hymenoptera Pteromalidae)rdquo Journal ofStored Products Research vol 41 no 4 pp 363ndash371 2005
[41] R Islam R I Khan S M Al-Reza Y T Jeong C H Song andM Khalequzzaman ldquoChemical composition and insecticidalproperties of Cinnamomum aromaticum (Nees) essential oilagainst the stored product beetleCallosobruchusmaculatus (F)rdquoJournal of the Science of Food and Agriculture vol 89 no 7 pp1241ndash1246 2009
[42] W S Nyamador G K Ketoh K Amevoin Y Nuto H KKoumaglo and I A Glitho ldquoVariation in the susceptibility oftwo Callosobruchus species to essential oilsrdquo Journal of StoredProducts Research vol 46 no 1 pp 48ndash51 2010
[43] H Kouninki T Hance F A Noudjou et al ldquoToxicity ofsome terpenoids of essential oils of Xylopia aethiopica fromCameroon against Sitophilus zeamais Motschulskyrdquo Journal ofApplied Entomology vol 131 no 4 pp 269ndash274 2007
[44] R Waliwitiya M B Isman R S Vernon and A RisemanldquoInsecticidal activity of selected monoterpenoids and rosemaryoil to Agriotes obscurus (Coleoptera Elateridae)rdquo Journal ofEconomic Entomology vol 98 no 5 pp 1560ndash1565 2005
[45] N S Iacobellis P Lo Cantore F Capasso and F SenatoreldquoAntibacterial activity of Cuminum cyminum L and Carumcarvi L essential oilsrdquo Journal of Agricultural and Food Chem-istry vol 53 no 1 pp 57ndash61 2005
