antimicrobial, antifungal and insecticidal investigations...

Vol 4(3) May-June 2005 179

Review Article

Antimicrobial, antifungal and insecticidalinvestigations on essential oils ó An overview

Gurdip Singh* and Sumitra MauryaChemistry Department

DDU Gorakhpur University , Gorakhpur - 273 009, Uttar Pradesh, India*Correspondent author, E-mail: [email protected]

Abstract

There is increasing interest both in the industry and in the scientificresearch for aromatic and medicinal plants because of their potential applicationsin medicine and plant disease control measures. The antimicrobial, antifungaland insecticidal properties of plant essential oils are well established againstwide spectra of organisms such as fungi, bacteria and insects. These propertiesare mainly due to many active phytochemicals including vitamins, flavanoids,terpenoids, carotenoids, coumarins, curcumines, etc. and hence, they are of greatimportance in food industry and offer the possibility to substitute natural forsynthetic preservatives and other products.

Keywords : Essential oils, Antimicrobial, Antifungal, Antibacterial, Insecticidal,Preservative.

IPC code; Int.cl.7 ⎯ C11B 9/00, A61K 35/78, A61P/00, A61P 31/04,A01N 65/00, A23L 3/3472

Introduction

Aroma chemicals present inleaves and flowers have been widely usedin aromatherapy since ancient times,suggesting that they have some beneficialhealth effects in addition to their pleasantodour. In addition to this, some herbs suchas rosemary and sage are used to producedrugs classified as pharmaceuticals,representing a significant part ofpharmaceutical market. The essential oilsrepresent a highly complex class of naturalproduct chemistry having well defined rolein the economic development of a country.Moreover, essential oils and extractsderived from plants have proved the

usefulness in controlling pre- and post-harvest fungal diseases (Fawcett et al,1970; Singh & Upadhayay, 1993; Lis-Balchin et al, 1995; Buchbauer, 1996;Singh, 1999; Singh et al, 1998a, 1998b,2003a). The oils affect the behaviourresponses of pests, kill or repel the pestswhich are harmful to human, animals orcrops (Singh & Pandey, 1982; Singh,1996a, 1996b; Singh et al, 1996). Thevolatile oils of some higher plants havebeen successfully exploited against anumber of storage and pathogenicmicroorganisms. These volatile oils havebeen found to be more efficacious thansome of the prevalent fumigants andhence, they are becoming popular in

recent years (Choudhary & Jain, 1991).Since these oils offer promisingcomplements or alternatives to traditionalsynthetic pesticides attempts tocharacterize the bioactive principles haverecently gained momentum in manypharmaceutical and food processingapplications. A chronological overview ofantimicrobial, antibacterial, antifungaland insecticidal investigations onessential oils has been presented in thisarticle.

Several factors may contribute todisparate observations on theantimicrobial potential of essential oils.Variability in qualitative and quantitativeestimates of activity has been ascribed todifferences in analytical techniques(Zaika, 1988; Mann & Markham, 1998).The volatility and poor solubility of mostessential oils are problematic, particularlywith methods that rely on diffusion ordilution of the test substance in amicrobiological medium. Generally theextent of inhibition of the oils could beattributed to the presence of an aromaticnucleus containing a polar functionalgroup (Farag et al, 1989). In addition,differences in composition related tovariety, agronomic practice andprocessing are also likely to influence theirproperties, since these factors contributeto both the profile and relativeconcentrations of active ingredients.

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Antibacterial investigations

Various microorganisms areknown to cause food spoilage and foodborne diseases in human beings. About200 diseases are caused by contaminatedwater, milk and other foods. The mostfrequently identified casual organisms are:Corynebacterium deptheriae(causing diphtheria of different organs),Escherichia coli (responsible forintestinal tract infection and diarrhoea),Staphylococcus aureus (responsiblefor obsess, sepsis and skin infections),Pseudomonas aeruginosa(responsible for hospital acquiredinfections), etc.

Oils for protecting food

A number of reports have beenpublished, on the action of Nigellasativa Linn. oil on different bacterialisolates. The oil has been reported to havea broad spectrum of activity against a

number of microbes. For example, invivo antibacterial effects of the essentialoil showed pronounced activity even in1:100 dilutions against several organismsthat included Staphylococcus albus, E.coli, Salmonella typhi, Shigellaniger and Vibrio cholera (El-Kamaliet al, 1998; Aggrawal et al, 1979).Generally speaking the oil was moreeffective against Gram positive than Gramnegative organisms. The effect wasmaximal when Bacillus subtilis wasused. Valeoro and Salmeron (2003) havegiven a description on the effects of addingessential oils of nutmeg, mint, clove,oregano, cinnamon, sassafras, sage, thymeand rosemary to carrot broth (used as agrowth substrate) on the growth of foodborne pathogen Bacillus cereus. Santoset al (1997) reported that the leafessential oil of Psidium guineense Sw.,P. pohlianum O. Berg andPilocarpus spicatus A. St. Hil showedstrong antibacterial activity (MIC 52.5-75.0 mg/mL) against Gram negativePseudomonas aeruginosa and againstGram positive Staphylococcus aereus.Psidium guineense demonstratedmarked activity (MIC 75.0 ±15.0 µg/mL)followed by Pilocarpus spicatus andP. pauciflorus A.St. Hil (MIC 1000.0±28.9 µg/mL). The essential oil from therhizomes of turmeric (Curcuma longaLinn. syn. C. domestica Val.) exhibitedgood to moderate activity against B.subtilis, S. aureus and C. diphtheriae(Garg & Jain, 2003). Souza et al (1998)reported that the essential oil from flowerheads of Egletes viscose Linn.displayed good antibacterial activityagainst resistant strains of S. aureus withminimum inhibitory concentration (MIC)ranging from 0.625-2.5µL/mL. Minija and

Thoppil (2003) reported that the volatileoil of Centella asiatica (Linn.) Urb.showed remarkable activity against E.coli. Bezic et al (2003) reported that theactivity of volatile oil of Achilleaclavennae Linn. was more pronouncedagainst Gram negative bacteria.

The activity of several parts ofPinus bruitia Ten., P. nigra Ait.,Juniperus oxycedrus Linn., Abiescilicica Ant. & Kotschy and Cedruslibani A. Rich were studied by Digarket al (1999). They reported that allextracts inhibited the growth of all bacteriaexcept E. coli. El-Sahawy et al (1998)have studied the essential oils of freshflowers, leaves and fruits of Murrayaexotica Linn., and reported strongantibacterial activity against E. coli, P.aeruginosa, S. aureus and Sarcinalutea. Moreover, Smith-Palmer et al(2001) reported that oils of bay, clove,cinnamon and thyme can act as potentinhibitors of Listeria monocytogenesand Salmonella enteritidis. The resultsof bacteriocidal and bacteriostaticconcentrations showed that the two Grampositive bacteria, S. aureus and E. coliwere more sensitive than Gram negativebacteria. This is in agreement with theresults of Prasad et al (1986). Thus, ingeneral, lower bacteriostatic andbactericidal concentrations are requiredfor basil essential oil against S. aureusand L. monocytogenes. It is notcompletely clear why Gram negativebacteria should be less susceptible, but itmay be associated with the outermembrane of Gram negative bacteria thatendows the bacterial surface with stronghydrophilicity and act as a strongpermeability barrier (Nikaido & Vaara,1985). De et al (1999) reported thatPsidium guineense

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clove, cinnamon, bishop’s weed, chili,horse raddish, cumin, tamarind, blackcumin, pomegranate seeds, nutmeg,garlic, onion, tejpat, cellary, cambodgehave potent antimicrobial activity againstBacillus subtilis (ATCC 6633), E. coli(ATCC 10536) and Saccharomycescerevisiae (ATCC 9763). Further, Lis-Balchin and Deans (1998) studied sixdifferent combinations of these differentessential oil mixtures for their possiblesynergistic activity against 13 food bornebacteria in vitro. The results indicatedthat there was no apparent synergisticactivity exhibited by any of the mixturesused. Delaquis et al (2002) reported thatthe essential oil of cilantro (coriander)was particularly effective against L.monocytogenes, likely due to thepresence of long chain (C

6-C

10) alcohols

and aldehydes. Mixing of fractions resultedin additive, synergistic or antagonisticeffects (Davidson & Parish, 1989). Didryet al (1993) observed synergistic activitybetween carvacrol and thymol. Lachowiczet al (1998) used an impedance methodto determine the time required forLactobacillus curvatus andSaccharomyces cerevisiae to initiategrowth in the presence of linalool andmethyl chavicol. Furthermore, the crudeessential oil was significantly moreeffective than any of the mixtures,suggesting that other components of basiloil contribute to antimicrobial activity. Lis-Balchin and Deans (1997) showed thatessential oils derived from commonbotanical sources varied widely incomposition and activity against L.monocytogenes. Most of theantimicrobial activity in essential oilsderived from spices and culinary herbsappear to derive from phenolic

compounds, while other constituents arebelieved to contribute little to theantimicrobial effects (Shelef, 1983;Martini et al, 1996; Nychas, 1998).Purified compounds derived fromessential oils, such as carvacrol, eugenol,linalool, cinnamic aldehyde and thymolinhibit a wide variety of microorganisms(Hulin et al, 1998). Individual essentialoils may contain complex mixtures of suchcompounds however, and little is knownabout the effect of interactions betweenindividual constituents on antimicrobialactivity.

Oils used for curing diseases

Various workers havedetermined the antibacterial activity ofmany oils and found rosemary oil highlyactive against a variety of bacteria. The oilsof Melaleuca leucadendron Linn.and Feronia limonia (Linn.) Swinglepossess significant antibacterial efficacy(Rao & Rao, 1976). Banerjee et al(1978) studied the antimicrobial activity

of the essentialoils ofC u r c u m aspp. Theantimicrobialactivity of theCinnamomumzeylanicumBreyn. leaf oilagainst threebacteria E.coli, S.aureus and P.aeruginosahas also been assayed (Hili et al, 1997;Smith-Palmer et al, 1998). Among theconstituents of C. zeylanicum,cinnamaldehyde possessed the bestantibacterial activity followed by cinnamylalchohol (Chang et al, 2001). Grover andRao (1976, 1978a, 1978b, 1979) havefound the oil of Daucus carota Linn.having considerable potential forcontrolling the growth of Bacillussubtilis and Salmonellatyphimurium. The essential oil ofBoswellia serrata Roxb. barkdisplayed significant inhibitory activityagainst Staphylococcus aureus, E.coli and Proteus mitrabilis (Kasaliet al, 2002). Regarding the antibacterialactivity of lemongrass oil, it is reported thatthe oil showed a high degree ofbactericidal activity against Gram positiveand Gram negative bacteria (Chao &Young, 2000; Onawunmi, 1989; Syed etal, 1995; Alam et al, 1994; Sharma etal, 2003; Saikia et al, 1999). It alsoinhibited the growth of Paenibacilluslarvae, the casual organism of Americanfoulbrood disease (AFD) of honeybees(Alippi et al, 1996). Ngassoum et al

Review Article

Callistemonlanceolatus

Cinnamomum zeylanicum


Review Article

(2003) reported that the essential oil offresh leaves of O. gratissimum Linn.and fruits of Zanthoxylumzanthoxyloides (Lam.) B. Zepernick& F.K. Timler are very effectiveantimicrobial agents.

Among all the oils, the essentialoil of cinnamon was most effective,followed by the essential oil of oreganoand thyme. The citronella oil was foundto be more effective than rose geraniumoil (Aggarwal et al, 2000). Citronella andgeranium oils as such were not effectiveagainst Gram negative bacteria but theircomponents, viz. citronellol and d-citronellol of citronella geranium oil andthose of citronellol and geraniol ofgeranium oil showed antibacterial activityagainst Gram negative bacteria, but muchless than Gram positive bacteria. Contraryto this Parcha et al (2003) have reportedsignificantly high activity of Dysoxylummalabaricum Bedd. oil against bothGram positive and Gram negative bacteria,like Shigella dysentriae andSalmonella typhi. Singh et al (1998a,

2001b, 2002c) reported that the volatileoils of some aromatic plants, viz. Lippiaalba (Mill.) N.E. Br. ex Britton & P.Wilson, Hyptis suaveolens Poit.,Alpinia cristata Griff., Callistemonlanceolatus DC., Eucalyptuscitridora Hook., Mentha piperitaLinn. emend. Huds, M. spicata Linn.emend. Nathh., Carum copticumHiern, Cuminum cyminum Linn.,Coriandrum sativum Linn. andApium graveolens Linn. as potentantimicrobial agents against humanpathogenic bacteria. Nagalakshmi et al(2003) reported that Chukrasiatabularis A. Juss. leaf oil exhibitedstrong antibiotic activity against Proteusvulgaris and Fusarium oxysporum.Melia dubia leaf essential oil exhibitedbacteriostatic activity againstPseudomonas aeruginosa,Escherichia coli and Klebsiellapneumoniae. The inhibitory activities ofboth essential oils were comparable withthat of respective standards. The strongantimicrobial activity of the essential oilof Hypericum maculatum Crantz hasbeen reported by Gudzic et al (2002)against S. aureus, E. coli, P.aeruginosa, Salmonella enteritidis,Klebsiella pneumoniae, Bacillussubtilis, B. enteritidis and Sarcinalutea.

Oils for protecting plants

Recently, Basim and Basim(2003) evaluated the antibacterial effectsof essential oil of Rosa damascena Mill.petals against three strains ofXanthomonas axonopodis pv.vesicatoria and reported that it may bea potential controlling agent in the

management of the disease caused by thisbacteria in tomato and pepper plants.

Antifungal investigations

Oils for protecting food and spices

Many publications havedocumented the antifungal activity ofLemongrass, Cinnamon, Clove, Palmaroseand Oregano oils against differentmicrobial species (Farag et al, 1989;Salmeron & Pozo, 1991; Patkar et al,1993; Sinha et al, 1993; Mishra & Dubey,1994; Paster et al, 1995; Hammer et al,1999; Inouye et al, 1998; Patnaik et al,1996; Chao & Young, 2000). Lemongrassoil is an effective post-harvest fungitoxicantof higher-order plant origin potentiallysuitable for protection of foodstuffs againststorage fungi (Mishra & Dubey, 1994). Invivo studies have shown that oregano oilis highly effective in controlling internalwheat fungi (Patker et al, 1993). Pattnaiket al (1996) reported that the palmaroseoil has some inhibitory activity against 12fungi; the response ranged from variousdegrees of susceptibility to total inhibition.It is reported that the response to this oilis dependent on the species of fungi. It isinteresting to note that the maincomponent of palmarose oil, geraniol(aliphatic alcohol), and geranial and neral(aliphatic aldehydes) main component oflemongrass oil, completely suppressed thegrowth of Aspergillus flavus andconsequently prevented the formation ofaflatoxin (Mahmound, 1994). Essentialoils containing aliphatic alcohols andphenols exhibited significant actionagainst A. aegyptiaceus, Penicilliumcyclopium and Trichoderma viride(Megalla et al, 1980). The inhibitoryEucalyptus citridora

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effects of clove and cinnamon oils on thegrowth and aflatoxin production by A.flavus have also been reported(Bullerman et al, 1977; Sinha et al,1993; Montes-Belmont & Carvajal, 1998).

A perusal of literature reveals thatthere are many reports on the fungitoxicnature of the Ocimum canum Sims oil(Bhargawa et al, 1981; Rao & Joseph,1971). The toxicity of the thermo-stablefungitoxic factor of this oil remained(Dubey et al, 1983) active during storageup to 390 days. Moreover, it successfullyprevented fungal deterioration ofFoeniculum vulgare Mill. (Fennel)and Coriandrum sativum (Coriander)during storage without showing anyphytotoxic effect on the treatedcommodities. Alvarez-castellanos et al(2001) reported that the essential oil offlower heads of garland Chrysanthemumhighly reduced (>80%) the growth of A.flavus, Pythium ultimum andAlternaria sp. The essential oil from theleaves of Murraya koenigii (Linn.)Spreng. has shown (Srivastava & Singh,2001) very good activity, especially against

Rhizoctonia bataticola andHelminthosporium oryzae with ED

50

0.1112% and 0.1214%, respectively. It hasalso been found (Pandey & Dubey, 1992)effective against Pythiumaphanidermatum and Rhizoctoniasolani.

Recently, Velluti et al (2003)reported that Cinnamon and Oregano oilsshowed inhibition on growth and FB

1

production by three different isolates ofFusarium proliferatum in irradiatedmaize grain. It was also found to beeffective against aflatoxin formation inmaize grain by A. flavus after 10 daysunder favourable conditions for mycotoxinproduction (Sinha et al, 1993). Oreganooil is also reported to inhibit the growthof A. flavus, A. ochraceus and A.niger (Paster et al, 1990, 1995).Mimica-Dukie et al (2003) reported thatthe method of distillation did slightly affectthe antifungal activity of the volatile oilsagainst fungi. According to Edris andFarrag (2003), individual constituents atdifferent ratios inhibited the growth of thefungi in a dose dependent manner.Menthol was found to be the individualaroma constituents responsible for theantifungal properties of peppermintessential oil, while menthone alone did notshow any effect at all doses. In case of basiloil, linalool alone showed no activity atall. Mixing the two components in a ratiosimilar to their concentrations in theoriginal oil was found to enhance theantifungal properties of basil oil indicatinga synergistic effect. Cinnamomumzeylanicum oil showed (Ouattara et al,1997) an inhibitory effect against meatspoilage organisms. Moreover, Singh andco-workers (2004a, 2005a) reportedPiper nigrum Linn. and Myristica

fragrans Houtt. (aril part) volatile oilsas 100% effective for Fusariumgraminearum whereas Zingiberofficinale Rosc. oil (Singh et al,2005b) is effective against F.oxysporum. In addition, Ajwain oil hasalso been reported (Singh et al, 2004b;Sridhar et al, 2003) to possess a broadspectrum of fungitoxic behaviour againstthe pathogenic fungi such as Aspergillusniger, A. flavus, A. oryzae, A.ochraceus, Fusarium monoliforme,F. graminearum, Penicilliumcitrium, P. viridicatum, P. madritiand Curvularia lunata isolated fromfood materials.

Oils for curing diseases

Most of the fungi cause diseasesin animals and plants. Several strains ofAspergillus niger are known to causemany important animal diseases such asmycotic abortion of sheep and cattle,pulmonary infection in birds andostomycosis, mycotic keratitis, allergy andrarely mycetoma commonly known asaspergillosis in humans (Rippon, 1998).The essential oils obtained fromCymbopogon martini (Roxb.) Wats.and C. citratus Stapf was studied againstsome fungi, viz. Cladosporium sp.,Aspergillus fumigatus, A. niger,Candida sp., Mucor, Trichophytonrubrum and T. viollacellium, causingdisease in human beings (Singatwadia &Katewa, 2001). The oil of C. citratus wasfound to be effective againstCladosporium sp., Aspergillusniger, and Mucor at lowerconcentrations, where as that of C.martini was more effective againstCandida sp., Aspergillus fumigatus

Zingiber officinale


Review Article

and Trichophyton rubrum comparedwith the oil of C. citratus. The volatileoil of C. pendulus (Nees ex Steud)Wats. exhibited strong activity againstMicrosporum gypseum (Bodin) Guiartand Trichophyton mentagrophytes(Robin) Blanchard. Both the oils inhibitedthe growth of fungi at equal concentrationin Trichophyton viollacellium(Pandey et al, 1996). Singh and co-workers (1995) have demonstrated thatCinnamomum zeylanicum bark oilhas fungitoxic properties against fungiinvolved in respiratory tract mycoses, suchas Aspergillus niger, A. fumigatus,A. nidulans and A. flavus. The activityof Japanese mint (Mentha arvensisLinn.) essential oil was studied and theminimum inhibitory concentrationsobserved for Candida albicans and C.neoformans (<1µL/mL), followed byM. gypseum (1.95µL/mL), S.schenckii (3.90µL/mL) and T. rubrum(62.5µL/mL), respectively (Rath et al,2001). Foeniculum vulgare volatileoil exhibited a broad range of antifungalactivity, inhibiting some nail infecting fungisuch as Aspergillus niger, A. flavus,

A. fumigatus, A. ustus, Candidaalbicans, Epidermophytonfloccosum, Microsporum audouini,M. canis, M. gypseum, M. nanum,Rhizopus nigricans, Trichophytontonsurons and T. violaceum (Patra etal, 2002). The efficacy of Curcumalonga oil against Colletotrichumgloeosporioides, Pestelotiapalmarium, Sphacelomacardamoni, Rhizoctonia solani andAspergillus spp. has also been reported(Saju et al, 1998). The oil has alsoexhibited good antifungal activity againstTrichophyton induced dermatophyte(Apisariyakul et al, 1995).

Oils for protecting plants

Singh et al (1992a) reportedfungitoxic activity of the citrus oil againstbroad spectrum of the fungi. They foundthat the oil of Citrus sinensis (Linn.)Osbeck showed strong fungitoxic activityagainst Helminthosporium oryzae at500 ppm and Fusarium oxysporum,F. vudum, F. monoliforme, Colletotricumfalcatum, Periconia atropurpurea,Ceratocystis paradoxa at 1000 ppmconcentration. However, the oil completelyinhibited the mycelial growth ofAspergillus niger, A. flavus,Rhizoctonia solani, Pythiumdebaryanum, P.aphanidermatum, Sclerotiumrolfsii, Colletotrichumgloeosporiodes, Curvularialunata and C. pallescens at2000 ppm concentration. Theoil has been found to be morepotent than the commonly usedsynthetic fungicides, viz. Phenylmercury acetate (e.g. Agrosan

GN), Carbendazin (e.g. Bavistin), Copperoxychloride (Blitox 50), Manizeb (e.g.Dithane M-45), Thiophenate methyl (e.g.Tropsil M) and Zineb (e.g. Dithane Z-78).The oil of Hyptis suaveolens Poit.showed lower minimum inhibitoryconcentration (MIC) as compared to thecommercial synthetic fungicides (Singhet al, 1992b). The essential oils ofDaucus carota Linn., Homalomenaaromatica Schott and Curcumazedoaria Rosc. was found to be moreefficacious in causing complete mycelialinhibition of Curvullaria pallescenceand Colletotrichum falcatum (Singhet al, 2002a).

Cuminaldehyde, an activeprinciple from the oil of Cuminumcyminum Linn., showed fungitoxicactivity against A. flavus and A. niger(Singh & Upadhayay, 1991). The leafvolatile oil of Tagetes erecta Linn.showed good activity against A. terreusand Colletotrichum falcatum (Singhet al, 2003b). Vidyasagar et al (2003)reported that volatile oil T. patula Linn.was found to be effective for Fusariumsolani and A. niger. Antifungal activityof T. minuta Linn. and its flavanoids hasbeen reported by Camm et al (1975)and Amat (1983). Among rare plants, the

Curcuma longa

Tagetes erecta

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volatile oils of Homalomenaaromatica and Lippia alba have beenreported to have significant activity (Singhet al, 2000a; Rao et al, 2000). The activityof L. scaberrima Sond. oil againstCandida albicans can be investigatedin the ranges<1300 ppm in liquid medium(Terblanche et al, 2001) whereasessential oil from the leaves of L.javanica Spreng. exhibited absoluteinhibition of the Alternaria alternate,Fusarium equiseti, Curvularialunata, Colletotrichum corchoriat 250 ppm and Macrophominaphaseolina, Botrydiplodiatheobromae at 750 and 1000 ppm,respectively (Chowdhury et al, 2003).However, among the Indian Curcumaspecies, C. zedoaria caused completemycelial inhibition againstColletotrichum falcatum at 2000 ppmand partial mortality for other tested fungi(Singh et al, 2002b, 2003c). The volatileoil of C. longa caused complete mycelialinhibition of C. falcatum, Fusariummonoliforme at 1000 ppm, C.pallescens, Aspergillus niger andF. oxysporum at 2000 ppm (Singhet al, 2001a, 2002b). According to Gargand Jain (2003), good to moderate activitywas observed using C. longa volatile oilagainst C. lunata, A. niger and A.fumigatus. The oil constituent turmeronehas demonstrated activity againstCandida albicans (Roth et al, 1998).Moreover, C. aromatica leaf oil causedcomplete mycelial inhibition of C.falcatum at 3000 ppm concentration andshowed inhibition for other tested fungi(Singh et al, 2002a). Recently, Wolkenet al (2002) worked on toxicity ofterpenes to spores and mycelium of

Penicillium digitatum and reportedcitral, geraniol, germanic acid and methylheptanone as most toxic terpenes. Skovicet al (2002) reported that the highest andbroadest activity was shown by thecarvacrol containing oils such asOriganum onites Linn. and Saturejathymbra Linn., while the oil of sage wasleast effective. The volatile oil of Nigellasativa was found to have excellentantifungal activity, particularly againstAspergillus species (Aggrawal et al,1979). Minija and Thoppil (2003)reported that the volatile oil of Centellaasiatica (Linn.) Urb showedremarkable activity against A. niger, R.oryzae, F. solani, C. albicans and C.musae. The volatile oil of Backhousiacitridora F. Muell possesses excellentactivity against a number of fungi such asA. niger, F. graminearum,Leptospaeria maculans,Microsporum gypseum,Trichophyton mentagrophytes, T.rubrum, and T. tonsurans (Wilkinsonet al, 2003). The volatile oil of Thymusstriatus Vahl exhibited strong inhibitoryeffect against all the test fungi. The oilcould be of better choice for developinglead compounds to control plant diseases(Couladis et al, 2004). Recently, Ozcanand Erkmen (2001) studied theantimicrobial activity of basilessential oil collected in Turkey. Theyfound the oil to be ineffective on S.cerevisiae, A. niger and Rhizopusoryzae, contrary to the findings of Dubeyet al (1989), Meena and Sethi (1994)and Prasad et al (1986). Thiscontradiction may be due to the differentchemotype of sweet basil or due todifferent test methods.

Insecticidal investigations

Some of the essential oils testedagainst various insects have shownpromising results. Yadava (1971) hasreported that emulsion of water andAcorus calamus Linn. oil showedstrong insecticidal activity againstBruchus chinensis. Su et al (1972)have found that non-volatile portions ofcitrus oil protected black-eyed peas fromCallosobruchus maculate infestations.The oils of Cymbopogon nardus(Linn.) Rendle, Cyathodia lyrata andTridax procumbens Linn. have alsoshown insecticidal activity. Many workershave reported the insecticidal activity ofoils of black pepper, Apiumgraveolens, Cuminum cyminum,Zanthoxylum alatum Roxb., Seseliindicum Wight & Arn., Luvungascandens (Roxb.) Buch-Ham. exWight, Nardostachys sp. andAzadirachta indica A. Juss.(Saraswati & Dixit, 1988; Singhmony etal, 1990; Dubey et al, 1991; Shukla etal, 1992; Dixit et al, 1992; Manuel et al,1988; Tom et al, 1979). Though nosubstantial work seems to have been doneon the structure activity relationship but afew available reports reveal that alkyl sidechain and free phenolic, hydroxy andmethylene dioxy groups (as in saffrole)are significant for showing ovicidal activity.The sweet smelling terpenoids attract theinsects on one hand and are ovicidal onthe other. Maini and Rajesis (1998)reported that the oils of sassafras, staranise and oregano showed 100% mortalityat 10 and 20 ppm against golden snails.The eggs and nymphs of White flies arealso affected by neem and citronella oils


Review Article

(Pant et al, 1993). Protectant potentialof essential oil of Tetradenia riparia(Hochst.) L.E. Codd against Zabrotessubfaciatus, infesting dried pin to beans,have been investigated by Weaver et al(1994). Adults of Corcyra cephalonicawhen exposed to volatiles of leaves of tulsior eucalyptus reduced egg yield in females(Maini et al, 1993). The essential oil ofSantolina chamaecyparissus Linn.reduced the survival of Nibparvathalugens after 72 hours and it was found tobe a good pesticide. The insecticidal andrepellent properties of nine volatileconstituents from essential oils againstPeriplaneta americana Linn., thebenzene derivatives (eugenol, methyleugenol, isoeugenol, safrole andisosafrole) were better (Ngoh et al, 1998)toxicants and repellants than themonoterpenes (limonene, cineol andp-cymene). Azadirachta indica seedderived azadirachtin shows nymphicidalactivity when injected into last instarnymphs of P. americana. Alder and Ubel(1985) reported the insecticidal anddevelopmental retardation effects afterfeeding pellets impregnating withMargosan-O18 (an azardirachtin-containing commercial formulation ofneem extract). The insecticidalconstituents of the Illicium verumHook.f. fruits were identified (Chang &Ahn, 2001), as phenylpropene (E)-anethole. The insecticidal activity of theconstituent was much more pronouncedin males than females. (E)-anethole, aconstituent of many aromatic plants suchas Pimpinella anisum Linn.,Clausena anisata (Willd.) Hook. f.ex Benth., Backhousia anisataVickery and Magnolia salicifolia(Sieb. et Zucc.) Maxim. (Harborne &

Baxte, 1993) possesses attractant effectsfor Diabrotica spp. (Lampman &Metcalf, 1987; Metcalf & Lampman, 1989)as well as insecticidal activity againstAcanthoscelides obtecus (Say)(Regnault-Roger & Hamraout, 1995),ovicidal activity against Daucus dorsaliseggs (Hendel) (Marr & Tang, 1992),adults of Drosophila melonogaster(M) and M. domestica third instarlarvae of Aedes aegypti (Marcus &Lichtenstein, 1979) and Sitophiluszeamais (Huang et al, 1997), fumiganttoxicity to adults of Lasiodermaserricorne (F), Sitophilus oryzae Land Callosobruchus chinenesis L(Kim & Ahn, 2001) and adults ofBlattella germanica L, M. domesticaand C. pipiens pallens (Coquillett) andMusca domestica (Chang, 2001). El-Nahal et al (1989) stated that the periodof exposure appears to be more importantthan dosage in influencing the efficiencyof the vapours of Acorus calamusessential oil to adults of five stored productinsect species. Similar results have beenreported for the toxicity of β-asarone toadults of S. oryzae and L. serricorne(Park, 2000). However, the adulticidalactivity of the phenylpropenes (E)-anethole and estragole and themonterpene (+)-fenchone against S.oryzae and L. serricorne is dependentupon both dose and exposure time. Butaccording to Chang and Ahn (2001), theadulticidal activity of (E)-anethole againstB. germanica varied according to thedose rather than the exposure time (Kim& Ahn, 2001). Chantraine et al (1998)evaluated insecticidal activity of 52Bolivian plant species has been evaluatedon Aedes aegypti larvae. Themonoterpenes E-anethol and E-nerolidol

were found to be the active principles ofthe most toxic essential oils. Singh et al(2000a, 2000b, 2002d, 2003b) workedon Odontotermes obesus Rhamb. andreported that the volatile oils ofCurcuma zedoaria, C. aromatica,Tagetes erecta, Coleus amboinicusLour and Homalomena aromaticahave a potent insecticidal behaviour.Traboulsi et al (2002) reported thatMyrtus communis Linn. oil was foundto be the most toxic, followed by those ofOriganum syriacum Linn., Menthamicrophylla C. Koch , Pistacialentiscus Linn. and Lavandulastoechans Linn. against the mosquitoCulex pipiens molestus Forskal(Diptera: culicidae). Amongmonoterpenes they found that (1R)-(+)-a-pinene and (1S)-(−)-α-pinene (LC

50 =

36-49mg/L) were most toxic whilementhone, 1, 8-cineole, linalool andterpineol (LC

50 = 156-194mg/L) were less

toxic. Recently, Kanat et al (2004)reported that essential oils of woodturpentine, thyme herb, cypress berry andstyrax are most effective in terms of meanmortality time against larvae of pineprocessionary moth (Thaumetopoeapitycampa Schiff.).

Conclusion

Essential oils are naturallyoccurring phytochemicals which havevarious applications such as naturalflavouring agent, pesticides, naturalperfumes and in aromatherapy. These areanalogous to synthetic chemical foodadditives, though so far only syntheticshave been specified. Therefore, anexecution of the appropriatestandardization for natural food additives

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is rapidly required, being important forhealth and food hygiene. Based on thisbackground many studies are in progress,most have still not yielded results and theyare difficult because many natural foodadditives are found in plant materials ascomplexes. Therefore, to begin with theincluded ingredients, particularly withmajor compounds and activities one mustclarify standards for the natural foodadditives.

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Researchers from Iowa StateUniversity reported that nepetalactone, theessential oil in catnip (Nepeta catariaLinn.) that gives the plant its characteristicodour, is about ten times more effective atrepelling mosquitoes than DEET - thecompound used in most commercialinsect repellents.

Aedes aegypti, which can carrythe yellow fever virus from one host toanother, is found in most parts of theUnited States. Researchers put groups of20 mosquitoes in a two-foot glass tube,half of which was treated withnepetalactone. After 10 minutes, only anaverage of 20 per cent - about fourmosquitoes - remained on the side of thetube treated with a high dose (1.0 percent) of the oil. In the low-dose test (0.1per cent) with nepetalactone, an average

Catnip oil repels mosquitoes moreeffectively than DEET

of 25 per cent - five mosquitoes - stayedon the treated side. The same tests withDEET (diethyl-m-toluamide) resulted inapproximately 40 percent to 45 per cent -eight-nine mosquitoes - remaining on thetreated side.

In the laboratory, repellency ismeasured on a scale ranging from +100per cent, considered highly repellent, to -100 per cent, considered a strongattractant. A compound with a +100 percent repellency rating would repel allmosquitoes, while −100 per cent wouldattract them all. A rating of zero meanshalf of the insects would stay on the treatedside and half on the untreated side. In tests,catnip ranged from +49 per cent to +59per cent at high doses, and +39 per centto +53 per cent at low doses. Bycomparison, at the same doses, DEET's

repellency was only about +10 per centin this bioassay. The nepetalactone is about10 times more effective than DEETbecause it takes about one-tenth as muchnepetalactone as DEET to have the sameeffect. Most commercial insect repellentscontain about 5 per cent to 25 per centDEET. Presumably, much less catnip oilwould be needed in a formulation to havethe same level of repellency as a DEET-based repellent. Why catnip repelsmosquitoes is still a mystery, it mightsimply be acting as an irritant or they don'tlike the smell. No animal or human testsare yet scheduled for nepetalactone. Ifsubsequent testing shows nepetalactone issafe for people, it can be commercializedas an insect repellent (http://www.zhealthinfo.com/catnip.htm).

antimicrobial, antifungal and insecticidal investigations...

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