23/3/2017 Antimicrobialefficacyoffiveessentialoilsagainstoralpathogens:Aninvitrostudy

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EurJDent.2013Sep7(Suppl1):S71S77. PMCID:PMC4054083
doi:10.4103/13057456.119078

Antimicrobialefficacyoffiveessentialoilsagainstoralpathogens:Anin
vitrostudy
NilimaThosar, 1SilpiBasak, 2RakeshN.Bahadure, 3andMonaliRajurkar2
1
DepartmentofPedodonticsandPreventiveDentistry,SharadPawarDentalCollege,DattaMegheInstituteofMedicalSciences(DeemedUniversity),
Wardha,Maharashtra,India
2
DepartmentofMicrobiology,JawaharlalNehruMedicalCollege,DattaMegheInstituteofMedicalSciences(DeemedUniversity),Wardha,Maharashtra,
India
3
Privatepractitioner,Nagpur,India
Correspondence:Dr.NilimaThosarEmail:drnthosar@rediffmail.com

Copyright:EuropeanJournalofDentistry

ThisisanopenaccessarticledistributedunderthetermsoftheCreativeCommonsAttributionNoncommercialShareAlike3.0Unported,whichpermits
unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalworkisproperlycited.

ThisarticlehasbeencitedbyotherarticlesinPMC.

Abstract

Objectives:
Thisstudywasaimedtofindouttheminimuminhibitoryconcentration(MIC)offiveessentialoilsagainstoral
pathogensandtofindouttheminimumbactericidalconcentration(MBC)andminimumfungicidalconcentration
(MFC)offiveessentialoilsagainstoralpathogens.

MaterialsandMethods:
TheantimicrobialactivitiesbydetectingMICandMBC/MFCoffiveessentialoilssuchasteatreeoil,lavenderoil,
thymeoil,peppermintoilandeugenoloilwereevaluatedagainstfourcommonoralpathogensbybrothdilution
method.ThestrainsusedforthestudywereStaphylococcusaureusATCC25923,EnterococcusfecalisATCC
29212,EscherichiacoliATCC25922andCandidaalbicansATCC90028.

Results:
Outoffiveessentialoils,eugenoloil,peppermintoil,teatreeoilexhibitedsignificantinhibitoryeffectwithmean
MICof0.620.45,9.0015.34,17.1231.25subsequently.MeanMBC/MFCforteatreeoilwas17.12
31.25,forlavenderoil151.00241.82,forthymeoil22.0012.00,forpeppermintoil9.7514.88andfor
eugenoloil0.620.45.E.fecalisexhibitedlowdegreeofsensitivitycomparedwithallessentialoils.

Conclusion:

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23/3/2017 Antimicrobialefficacyoffiveessentialoilsagainstoralpathogens:Aninvitrostudy

Peppermint,teatreeandthymeoilcanactasaneffectiveintracanalantisepticsolutionagainstoralpathogens.

Keywords:Antimicrobialactivity,essentialoils,oralpathogens

INTRODUCTION
Thespreadofdrugresistantpathogensisoneofthemostseriousthreatstosuccessfultreatmentofmicrobial
diseases.Essentialoilsandotherextractsofplantshaveevokedinterestassourcesofnaturalproducts.[1]Essential
oilsalsocalledvolatileoils,arearomaticoilyliquidsobtainedfromplantmaterialssuchasflowers,buds,seeds,
leaves,twigs,bark,herbs,wood,fruitsandroots.Anestimated3000essentialoilsareknown,ofwhich300are
commerciallyimportantinthefragrancemarket.[2]Theantimicrobialactivityofessentialoilsisduetoanumberof
smallterpenoidsandphenolcompounds.[3]Severaloftheseareclassifiedasgenerallyrecognizedassafe.[4]
Essentialoilssuchasteatreeoil,lavenderoil,thymeoil,peppermintoilandeugenoloilhavebeentraditionally
usedbypeopleforvariouspurposesindifferentpartsoftheworld.

Therootcanalenvironmentafterchemomechanicaltreatmentbecomesunfavorableformicroorganismsthereis
reducedoxygentension,limitednutrientavailabilityandantimicrobialagentsthatactasdrivingforcesinsurvival
balanceofbacteriaintherootcanalsystem.[5]Rootcanaldentinaltubulesharbormicroorganismsalsobacterial
biofilmmaybepresentattheapicalportionofrootcanalandextraradicularregions.[6]Therefore,irrigationwitha
broadspectrumantisepticsubstancesandinterappointmentintracanalmedicationhasbecomeastandardregimenin
rootcanaltherapy.Manyspeciesandherbsexertantimicrobialactivityduetotheiressentialoilfractions.For
thousandsofyearscloveoil(eugenol)hasbeenusedindentistry.Eugenolhasbeenusedtopicallyindentalpractice
torelievepainarisingfromavarietyofsources,includingpulpitsanddentinalhypersensitivity.Interestingly,
eugenolexhibitsirritantactioninadditiontoitsanalgesiceffectasfoundincertainstudies.[7]Soinsearchofplant
essentialoilspossessingantimicrobialactivity,thisstudywasundertakentodetermineinvitro,antimicrobialaction
ofeugenol,thymeoil,teatreeoil,peppermintoilandlavenderoilagainstfourpathogenicoralmicroorganisms.

Inthisstudy,itwasaimedtodetectinvitroantimicrobialefficacyoffiveessentialoilsagainstoralpathogens.
Firstly,itwasaimedtofindouttheminimuminhibitoryconcentration(MIC)offiveessentialoilsagainstoral
pathogens.Secondly,itwasaimedtofindouttheminimumbactericidalconcentration(MBC)andminimum
fungicidalconcentration(MFC)offiveessentialoilsagainstoralpathogens.

MATERIALSANDMETHODS
Weusedfiveessentialoilsasfollowing:

Teatreeoil,lavenderoil,thymeoil,peppermintoil,andeugenoloil.

Essentialoils
Fiveessentialoilssuchasteatreeoil,lavenderoil,thymeoil,peppermintoilandeugenoloilwereobtainedfrom
Arromatantra,Mumbai,India.Theseoilswereselectedbasedontheliteraturesurveyandtheiruseintraditional
medicine.Qualityofoilswasascertainedtobepure.

Testorganism
MicroorganismswereobtainedfromtheDepartmentofMicrobiology,JawaharlalNehruMedicalCollege,Wardha,
Maharashtra,India.ThestrainsusedforthestudywereStaphylococcusaureus(ATCC25923),Enterococcus
fecalis(ATCC29212),Escherichiacoli(ATCC25922)andCandidaalbicans(ATCC90028).Stockculturesof
bacteriawereusedforthestudy.

MICandMBC/MFCdetermination

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23/3/2017 Antimicrobialefficacyoffiveessentialoilsagainstoralpathogens:Aninvitrostudy

TheantimicrobialactivitiesbydetectingMICoffiveessentialoilssuchasteatreeoil,lavenderoil,thymeoil,
peppermintoilandeugenoloilwereevaluatedagainstfourcommonoralpathogensbybrothdilutionmethod[
Figure1].[8]MBCwasdetectedbysubculturingontobloodagarfromthetubeshowingnoturbidity(i.e.,MIC)
andthenexttubetoit.Thebloodagarplateswereincubatedat37Covernightfor1820handonthenextdaythe
readingsweretaken.ForMFC,subculturesweredoneonSabouraud'sdextroseagarplatesfollowingthesame
procedure[Figures25].Variousconcentrationsoffiveessentialoilsusedagainstoralpathogenswereindoubling
dilutions[Table1].Resultswereanalyzedstatisticallybyusingtheonewayanalysisofvariance.

Figure1
MinimuminhibitoryconcentrationofthymeoilforStaphylococcusaureus

Figure2
MinimumbactericidalconcentrationforEnterococcusfecalis

Figure5
MinimumfungicidalconcentrationforCandidaalbicans

Table1
Differentconcentrationsofessentialoilsagainstcommonoralpathogens

Figure3
MinimumbactericidalconcentrationforEscherichiacoli

Figure4
MinimumbactericidalconcentrationforStapholococcusaureus

RESULTS
Outoffiveessentialoils,eugenoloil,peppermintoil,teatreeoilexhibitedsignificantinhibitoryeffect[Table2]
withmeanMICof0.620.45,9.0015.34,17.1231.25subsequently.Therewasnostatisticalsignificant
differenceinfiveessentialoils(F=1.61,P=0.221)[Table3].MeanMBC/MFCforteatreeoilwas17.12
31.25,forlavenderoil151.00241.82,forthymeoil22.0012.00,forpeppermintoil9.7514.88andfor
eugenoloil0.620.45(F=1.30,P=0.312)[Table4].Eugenolshowedantimicrobialactivityatthelowest
concentrationcomparedwithallessentialoilsi.e.,forC.albicans:MIC,MFC:0.1l/ml,forS.aureus:MIC,
MBC:0.4l/ml,forE.coliandforE.facelis:MIC,MBC:1l/mlrespectively[Figures25].

Table2
MICandMBC/MFCoffiveessentialoils(l/ml)

Table3
MICoffiveessentialoils

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23/3/2017 Antimicrobialefficacyoffiveessentialoilsagainstoralpathogens:Aninvitrostudy

Table4
MBC/MFCoffiveessentialoils

DISCUSSION
TeatreeoilisthevolatileessentialoilderivedmainlyfromtheAustraliannativeplant,Melaleucaalternifolia.Tea
treeoiliscomposedofterpenehydrocarbons,mainlymonoterpenes,sesquiterpenesandtheirassociatedalcohols.
Terpenesarevolatile,aromatichydrocarbonsandmaybeconsideredpolymersofisoprene,whichhastheformula
C5H8.[9]Antibacterialactivityinliteratureappearedfrom1940to1980.[10,11,12,13]Fromtheearly1990sonward,
manyreportsdescribingtheantimicrobialactivityofteatreeoilappearedinthescientificliterature.[9]

Antimicribialactivityofteatreeoilisduetoterpinen4ol,terpineoland1,8,whichcauseleakageof260nm
lightabsorbingmaterialandrendercellssusceptibletosodiumchloride.[14]Thus,teatreeoilcauseslysisandthe
lossofmembraneintegrityandfunctionmanifestedbytheleakageofionsandtheinhibitionofrespiration.[9]
AntimicrobialresistantisolatesofS.aureus,[15,16]C.albicans,[17]andE.faecium[18,19]havebeenfoundtohave
invitrosusceptibilitiestoteatreeoil.Inthisstudy,antimicrobialeffectofteatreeoilforC.albicans(MIC:0.5
l/ml,MFC:0.5l/ml)wasfoundatthelowestconcentrationfollowedbyS.aureus(MIC:1l/ml,MBC:2l/ml)
andE.coli(MIC:2l/ml,MBC:2l/ml),lastlyE.faecalis(MIC:64l/ml,MBC:64l/ml).

Muchofpurelavenderoil(LavandulaofficianalisL.angustifolia[Miller]orL.veraLabiatae/Lamiaceae)comes
fromBalkans.Francestillproducesthefinestquality,butproductionthereistumbledwiththeadventofthehybrid.
Essentialoiloflavenderisobtainedfromthefloweringtopsbysteamdistillationmethod.Itsprincipalconstituents
includemonoterpens,Oxides,linalylandgeranyleesters,geraniol,linalooletc.,Ithasvarietyofproperties,which
includesthatitisantidepressant,hypotensive,soothing,alleviatesstress,anxietyandgeneraldebility.Itis
antisepticandantiinflammatoryforcolds,fluandsinusitisandthroatinfections.Itisbalancing,antiseptic,anti
inflammatoryandregenerativesoothesacne,eczema,dandruff,hairloss,headlice,diaper/nappyrash,sunburn,
insectbitesandboilsrelievesAthlete'sfootandherpessimplex.Ithascleansingandcalmingeffecthelpscolic,
dyspepsia,indigestion,flatulenceandgastroenteritis.Itissedativeanddecongestantlowersbloodpressurereduces
palpitations.Ithasmusculoanalgesicandantiinflammatoryeffectinneuritis,neuralgia,muscularsprains,cramps,
achesandpains.Itisalsousedininhalations,vaporizers,compresses,bathapplicationormassage.[20]Inthestudy
ofZuzarteetal.,2009[21]antifungalactivityoflavenderoilwasfoundwithMICvaluesof0.320.64l/ml.Inthe
studyofZuzarte(2011)[22]candidaspecies,werefoundtobesensitivetolavenderoilwithMICof0.64l/ml.In
ourstudy,itwasachievedwithahigherconcentrationi.e.,8l/mlforC.albicans.S.aureusandE.faecaliswere
susceptibleattheconcentrationofMIC:32l/mlandMBC:64l/mlrespectively.E.coliwasleastsusceptible
withMIC:128l/mlandMBC:512l/ml.

ThenamethymeactuallycomesfromtheGreekwordthymos,meaningsmallbecauseofthefragranceofthe
plant.Thymebelongstoover300speciesofhardy,perennialherbaceousplantsandshrubsthatarenativeto
Europe,particularlyaroundMediterranian.ItisoneoftheHippocrates400simpleremedies.Essentialoilofthyme
(ThymusSpp,T.citriodorits,T.vulgarisLabiatae/Lamiaceae)isobtainedfromtheleavesandfloweringtopsby
steamdistillationmethod.Itsprincipalconstituentsinclude2040%thymolandcarvacrolwithborneol,cineol,
linalool,menthone,Bcymene,pineneandtriterpenicacid.Thymeoilisatonicstimulantandstomachicand
digestiverelievesgastritis,enterocolitisandmouththrush.Itisusefulforrespiratoryinfections,asthmaand
bronchitis.Itiseffectivefortreatingswellingprovokedbygoutorrheumaticproblems,forjointpains,backache
andsciatica.Thymeoilisalsousefulforurinaryandvaginalinfections,endometritis(candida),prostratesand
vaginitis.[20]Thymeoilexhibitsantibacterialactivityandhasbeenusefulindentalpractice.[23]Acomponentof
thyme,knownasthymol,appearstoinhibitgrowthoforalpathogensinthemouthandincombinationwithother
essentialoils,mayreducedentalcaries.[24,25]Inpatientswithorthodonticbrackets,adentalvarnishcontaining
thymolreducedtheproportionofStreptococcusmutansinsupragingivalplaquenearthebracket.[26]Thymolisone
oftheessentialoilswithantibacterialeffectsfoundinListerine.[27]InthestudyofGisleneetal.,2000[28]Hiliet
29 30
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23/3/2017 Antimicrobialefficacyoffiveessentialoilsagainstoralpathogens:Aninvitrostudy

al.,1997[29]andNzeakoetal.,2006[30]thymeandcloveoilpossessedantimicrobialactivityagainstS.aureus,E.
coliandC.albicansatvariousconcentrationoftheextracts.Inourstudy,antimicrobialsusceptibilityinorderof
sequenceforthymeoilwasE.coliwithMIC:2l/ml,MBC:8l/ml,C.albicanswithMIC,MFC:16l/ml,E.
faecaliswithMIC,MBC:32l/mlandS.aureuswithMIC,MBC:32l/mlrespectively.

Essentialoilofpeppermint(MenthapiperitaLamiaceae/Labiatae)iscultivatedonawidescaleinEurope,USA
andJapan.Itisextensivelyusedintoiletry,foodandpharmaceuticalindustries.Avarietyofproductsrangingfrom
toothpastes,mouthwashesanddigestivetabletstosweets,icecreamandliquorsareflavoredwithpeppermint.
Essentialoilofpeppermintisobtainedfromtheleavesbysteamdistillationmethod.Itsprincipalconstituents
includemonoterpinicalcoholsmainlymenthol(3848%),ketonesmainlymenthones(2030%),somemonoterpens
andoxides.Itisagoodantiseptic,antibacterialandantiviral.Ithaslight,clean,refreshingaromaandisagood
insectrepellant.Ithasstimulatingandstrengtheningeffectintreatmentofshock,helpfulforneuralgiaandreliefof
generaldebility,headachesandmigraines.Ithasantisepticandantispasmodiceffectinreducingmucusand
relievingcoughs,sinusitis,throatinfections,colds,flu,asthmaandbronchitis.Itisalsousedininhalations,bathsor
applications.Ithasgotcoolingandcleansingeffectsoothesitchyskin,relievesinflammation.Ithassoothingand
antispasmodiceffectrelievesacidity,heartburn,diarrhea,indigestionandflatulence,alsoeffectivefortravel
sicknessandnausea.Ithascoolingeffectincaseofvaricoseveinsandhemorrhoids.[20]Peppermintoilmakesthe
mouthfeelfreshandofcourse,makestheformulatastegood.Peppermintoilcanalsoincreasesalivation,whichis
usefulbecausedrymouthmayresultinhalitosis.[31]Inourstudy,antimicrobialeffectwasachievedatthe
concentrationof0.5l/mlforC.albicansandattheconcentrationof32l/mlforE.coli,S.aureus,E.faecalis(32
l/ml).

ThecloveplantgrowsinwarmclimatesandiscultivatedcommerciallyinTanzania,Sumatra,theMaluku
(Molucca)IslandsandSouthAmerica.Thetallevergreenplantgrowsupto20mandhasleatheryleaves.The
clovespiceisthedriedflowerbudofEugeniacaryophyllataspecies.Essentialoilsareobtainedfromthebuds,
stemsandleavesbysteamdistillation.Thebudsorclovesarestronglyaromatic.Clovebudsyieldapproximately
1520%ofavolatileoilthatisresponsibleforthecharacteristicsmellandflavor.Thebudalsocontainsatannin
complex,agumandresinandanumberofglucosidesofsterols.Theprincipalconstituentofdistilledclovebudoil
(6090%)iseugenol(4allyl2methoxyphenol).Theoilalsocontainsabout10%acetyleugenolandsmallquantities
ofgallicacid,sesquiterpenes,furfuralandvanillinandmethylnamylketone.Otherconstituentsinclude
flavonoids,carbohydrates,lipids,oleanolicacid,rhamnetinandvitamins.[32]

Eugenoliswidelyusedandwellknownforitsmedicinalproperties.Traditionalusesofcloveoilincludetheusein
dentalcare,asanantisepticandanalgesic.[33]Itisactiveagainstoralbacteriaassociatedwithdentalcariesand
periodontaldisease[34]andeffectiveagainstalargenumberofotherbacteria[35,36,37,38]andvirus.[39]Previous
studieshavereportedbiologicalactivitiesofeugenolincludingantifungal,[40,41,42]anticarcinogenic,[43]anti
allergic,[44,45]antimutagenicactivity,[46]antioxidant[47]andinsecticidal[48]properties.Inourstudy,eugenoloil
showedantimicrobialactivityatthelowestconcentrationagainstallorganismscomparedwithallessentialoilssuch
asforC.albicanswithMIC,MFC:0.1l/ml,forS.aureuswithMIC,MBC:0.4l/ml,forE.coliandforE.
faceliswithMIC,MBC:1l/mlrespectively.

CONCLUSION
Hence,thisstudyconcludesthatapartfromtraditionaluseofeugenol,antibacterialeffectsofessentialoilssuchas
peppermintoil,teatreeoil,thymeoilalsocanprovideaneffectiveintracanalantisepticsolutionagainstoral
pathogens.

Footnotes
SourceofSupport:Nil.

ConflictofInterest:Nonedeclared

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