Rev. bras. farmacogn. vol.26 número3

w ww . e l s e v i e r . c o m / l o c a t e / b j p

Original

Article

Exudates

used

as

medicine

by

the

“caboclos

river-dwellers”

of

the

Unini

River,

AM,

Brazil

–

classification

based

in

their

chemical

composition

João

Henrique

G.

Lago

_,

_Jaqueline

_Tezoto

_,

_Priscila

_B.

_Yazbek

_,

_Fernando

_Cassas

_,

Juliana

de

F.L.

Santos

_,

_Eliana

_Rodrigues

a_{DepartmentofBiologicalSciences,CentrodeEstudosEtnobotânicoseEtnofarmacológicos,UniversidadeFederaldeSãoPaulo,Diadema,SP,Brazil} b_{DepartmentofExactSciencesandEarth,UniversidadeFederaldeSãoPaulo,Diadema,SP,Brazil}

c_{Coordenac¸ãoemCiênciaeTecnologia,UniversidadeFederaldoMaranhão,SãoLuís,MA,Brazil}

a

r

t

i

c

l

e

i

n

f

o

Articlehistory: Received30June2015 Accepted14March2016 Availableonline28March2016

Keywords: Amazonforest Ethnobotany Medicinalplants Ethnopharmacology Exudatesphytochemistry

a

b

s

t

r

a

c

t

Althoughtheuseofexudatesintraditionalmedicinehasbeencommonlyobservedduring ethnophar-macologicalsurveys,fewrecordshavebeenmadeconcerningthescientificmeritsoftheseproducts.The aimofthisstudywastodocumentethnopharmacologicaldataandtoclassifyexudatesusedasmedicine bythe“caboclos”river-dwellersfromtheUniniRiverofAmazonas,Brazil,onchemicalanalysesbasis. Usinganethnographicapproach,indicatedplantsandtheirrespectiveexudateswerecollected, identi-fiedandincorporatedintoherbariumoftheNationalInstituteofAmazonianResearch.Toclassifythese exudates,plantmaterialwasextractedusingmethanol,andobtainedextractswereanalyzedbyNuclear MagneticResonanceandmassspectrometryaimingidentificationofmaincompounds.Fifteenexudates wereindicatedby“caboclos”river-dwellersashomeremedies;amongtheirtherapeuticuses, inflamma-toryprocesses,culture-boundsyndromesandrespiratorydiseasesaremostprominent.Basedontheir solubilityandchemicalsclasses,fifteenexudateswereclassifiedinto:latex(7),resins(5),sap(1),gum (1),oleoresin(1);andelevenofthemhavenotbeenmentionedonpharmacologicalliteratureuntilthis moment.Theobtainedresultsmaycontributetochemical/pharmacologicalapplicationofexudatesfrom thesespecies,severalofwhichhavebeenclassicallyusedinBrazilianfolkmedicine.

©2016SociedadeBrasileiradeFarmacognosia.PublishedbyElsevierEditoraLtda.Thisisanopen accessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction

Althoughtheuseofexudatesintraditionalmedicinehasbeen commonly observed during ethnopharmacological surveys, few recordshavebeenmadeconcerningthescientificmeritsofthese products.InBrazil,theuseofresinhasbeennotedamongthe eth-nicgroupsofKapoor(Balée,1994),Paumari(Pranceetal.,1987) andWaimiriAtroari(Millikenetal.,1992).Apossibleexplanation forthegapinrecordsisthedifficultyinidentifyingtheexudates, sincetheseexudatesresembleoneanotherinphysicalappearance. However,theircharacterizationandmeansofdistinctionhavebeen facilitatedinrecentyears,asaresultoftechnologicaldevelopments in chemistry, molecularbiology andmicroscopy. Water-soluble exudatesinclude gums,whicharecomposedofpolysaccharides

∗ Correspondingauthor.

E-mail:e.rodrigues@unifesp.br(E.Rodrigues).

and are secreted by wells;and saps, which consistof polysac-charides and amino acids,as described byLangenheim (2003). Accordingtothesameauthor,fat-solubleexudatesincluderesins andoleoresins,which arecomposedofterpenoidsandphenolic compoundssecretedbyresiniferouschannels,cavities,trichomes andepidermalcells;oilsarecompoundsoffattyacidsandglycerol; andlatexisacomplexmixtureofterpenoids,phenoliccompounds, proteinsandcarbohydratesanditissecretedbylaticifers.

Not only exudates are poorly studied from the ethnophar-macological standpoint,but, tothebest ofour knowledge,few pharmacological and chemical studieshave beenconducted on exudates.Ontheotherhand,thereareAmazoniancultureswhose medicinal uses of exudates need to be identified and docu-mented before they are lost due to the rapid introduction of syntheticmedicines.Therefore,theaimofthisstudyistodocument ethnopharmacologicaldataandclassify,onthebasisofchemical analysis,theexudates(aslatex,resin,oleoresinandsoon)used asmedicineby“caboclos”river-dwellersfromtheUniniRiverof Amazonas,Brazil.

http://dx.doi.org/10.1016/j.bjp.2016.03.001

Materialsandmethods

Ethnopharmacologicaldata

Theexudatesanalyzedinthisstudywereindicatedbytwelve healers of “caboclos” river-dwellers living in two communities alongtheUniniRiverofAmazon,Brazil(Fig.1),during11months of fieldwork, between April 2008 and January2012 by one of theauthors (JFLS). For local healing experts selection, a snow-ball sampling, as described by Bernard (1988), was performed by consultation of local inhabitants of riverine communities. Ethnographic techniques and methods were applied, including participantobservation, fielddiaries, informaland unstructured interviews (Bernard, 1988; Foote-Whyte, 1990). During inter-views,thefollowingdatasheetswereadministered:interviewee personalinformation,ethnopharmacologicalsurvey(ingredients, uses,parts used,modeof preparation,and contraindications of plantsandanimalsusedfortherapeuticpurposes)andplant collec-tion(popularname,habit,timeofflowering/fruiting,organoleptic and morphological aspects) (Santos et al., 2012).The exudates indicatedbyintervieweesandtheirrespectivevegetablematerials werecollectedduringfieldworkatTerraNova(01◦₄₁′_S,61◦₄₉′_W)

andTapiira(01◦₄₆′_S,62◦₁₃′_{W)communities(Fig.1),theywere}

identified by Mr. José Ramos, plant taxonomy technician, and incorporatedintotheherbariumoftheNationalInstituteof Ama-zonianResearch.Priortothefieldwork,allnecessarypermitswere obtainedforthestudy,includingapermitaccesstothe Conserva-tionUnits,inordertoensurecollectionofsamples,transportof biologicalmaterialandaccesstoassociatedtraditionalknowledge, andprior informed consentofinformants (SISBIONo. 16805-2, CGEN/MMANo.47/2009andCEP-UNIFESP/EPM,No.1354/08).

Exudatesextraction

Liquidexudates(latex,oleoresinandsap),wereobtainedfrom trunksoftheirrespectivetreesbyincisionsutilizingmacheteor knife,dependingonthethicknessofthebarks.Apointedmetal can-nulawasadaptedinthetrunkofeachtreeforcollectionofexudates.

Thesolidsandsemi-solidsexudates(resinandgum,respectively), werestuckontreetrunks;thereforetheywerecollected manu-ally.Allexudateswereplacedinamberglasses,properlylabeled withdateandnumericcodescorrelatingthemtothetreesfrom whichtheywerecollected.Theseglasseswerekeptinastyrofoam boxuntiltheyreachthelaboratorywheretheirextractswere pre-paredforchemicalanalysis.Allexudatesweredriedaftercollection, beforethepreparationoftheextracts.

Preparationofextractsandchemicalanalysis

Solubilityofobtainedexudateswasevaluatedusing10mgof eachplantmaterialand1mlofH2Oorhexanesinatest-tube.Then, exudateswereextractedusingMeOH(50mlofsolventtoeach10g ofplantmaterial)inasonicatedbathatroomtemperatureduring 20min.Aftersolventevaporationunderreducedpressure,obtained crudeextractswereanalyzedby1_Hand13_CNMR(nuclear mag-neticresonance)spectroscopyinaBrukerAvance300spectrometer (300MHz to 1_{H and 75MHz to} 13_{C nuclei, respectively) using} CDCl3,CD3OD,(CD3)2SO,(CD3)2OorD2Oassolventsandinternal standard.CrudeextractswerealsoanalyzedbyHPLC/ESIMS (liq-uidchromatography/electrosprayionizationmassspectrometry) usingaBrukerDaltonicsequipmentEsquire3000Plus.HPLCsystem wascoupledwithaZorbax-C18(250mm×4.6mm,3.55m,Agilent, USA)columnat40◦_{C.SolventsH2Oandacetonitrile(CH3CN)were}

used,startingwith15%ofCH3CN(0–20min),increasingto100% (20–25min),isocratic(5min),anddecreasingto15%(30–32min), and isocratic(3min) atflow rateof1ml/min.Injectionvolume was3␮landUVdetectionat352nmand280nm.ESIMSspectra (lowresolution)wererecordedinfullscanandproductionscan modes(argonCID).Ionsourcewassetasfollows:nebulizergas=3 l/min,desolvationgas=15 l/min,DL=150◦_{C, heatblock=300}◦_C

andvoltage=3.5kV.GC/EIMS(gaschromatography/lowresolution electronicimpactmass spectrometry) weremeasuredin a Shi-madzuGC-17Achromatographequippedwithacapillarycolumn DB-5(crosslinked5%phenylin95%silicone-30m,0.32mm,I.D., 0.25␮mfilmthickness)interfacedwithaMS-QP-5050Amass spec-trometer.Temperatureprogrammingwasperformedfrom150to

Tapiira

Terra Nova

Roraima

Unini River Negro River

Manaus

154 Km W Amazon State

Brazil

E N

S W

E S

10.8 km

U

N

I

N

I

R

I

V

E

R

N

Figure1. UniniRiverlocationintheAmazonforestbiome,AmazonState–Brazil(mapontheright).LocationoftwoUniniRivercommunities,wheretheexudateswere collectedduringfieldwork:TerraNova–01◦₄₁′_S,61◦₄₉′_{WandTapiira–01}◦₄₆′_S,62◦₁₃′_W(left).

300◦_Cat15◦_{C/min,thenisothermalat300}◦_{Cfor5min.Injector}

anddetectortemperatureswere150◦_Cand320◦_{C,respectively,}

andheliumwasusedasthecarriergas.Identificationofcompounds wasconductedbyanalysisofindividualNMRandMSspectra(low resolution).Obtained data werecompared withthose recorded toauthenticsamples(␣-and␤-amyrin,sitosterol, stigmasterol, quercetin, quercitrin, catechin, lupeol, friedelin, betulinic acid, copalicacid,oleanolicacid,ursolicacid,palmiticacid,brasiliensic acidandisobrasiliensicacidwereavailableatourlaboratorywhile lysinandtryptophanwereobtainedfromSigma–Aldrich)and/or withthoseavailableintheliterature.

Bibliographicsurvey

Fromplantspeciesidentified,wesearchedscientificdatabases (PubmedandScopusJune/2015)todeterminewhethertheir exu-dateshadbeentargetsofpreviouspharmacologicalstudies.

Results

Allthetwelveinterviewees(sevenfemaleandfivemale)were bornin theMiddle NegroRiverregionandtheyare descended ofinhabitantsfromAmazonasandCearáStates.Theycall them-selvesintofollowingcategories:knowledgeableofnaturaldrugs (sicinterviewees),healer(3), midwife(2)and ‘desmintidor’(an expert in massage techniques for bone dislocationand muscle straintreatments)(1).Theyreportedhavinglearnedthehealing practiceswithparents,relatives,friends,neighbors,andoftenasa self-interestresult,asitisthecaseofmidwivesand‘desmintidores’ (Santosetal.,2012).

AsshowedonBox 1, “caboclos”river-dwellersindicated fif-teenexudates.MembersoftheBurseraceaefamilywerethemost frequentlyidentified,totalingfourspecies.Theirtherapeuticuses weregivenastheirlocaltermsand theyaresignalizedinitalic throughoutthis paper.Amongtheirtherapeuticuses, inflamma-toryprocesses,culture-boundsyndromesandrespiratorydiseasesare

Box1:The15plant’sexudatesusedasmedicinebythe‘caboclos’river-dwellersoftheUniniRiver,AM;their pharma-cologicaldatapresentinscientificliterature;theirrespectiveclassificationsbasedonourchemicalanalysis,andthe maincompoundsidentified.

Family Scientificname/vernacular name(voucher)

Medicinaluse(localterm) Pharmacologicalstudies (plantpart/exudatestudied)

Exudates(solubility) Mainidentified compounds**

Apocynaceae CoumamacrocarpaBarb. Rodr./leite-de-sorva(ER097)

Weaknessinthelung – Latex(fat-soluble) –

Araceae Philodendronsolimoesense

A.C.Sm./cipó-ambé (ER2000)

Cataract – Latex (fat-soluble) –

alkylphenol, steroids

3-Octadecenyl-phenol, sitosterol, stigmasterola,b,d Bignoniaceae Tynanthuspanurensis

(BureauexBaill.)

Sandwith/cipó-cravo(ER2003)

Improvingmemoryand

calming

– Sap(water-soluble) –

Burseraceae Protiumamazonicum

(Cuatrec.)Daly/breu-branco (JFLS413)

Headache,strokeand diseaseoftheair

– Resin(fat-soluble)–

triterpenes

␣-Amyrin,␤-amyrina,b,d

Protiumaracouchini(Aubl.) Marchand/breu-preto (JFLS404)

Headache,strokeand diseaseoftheair

– Resin(fat-soluble)–

triterpenes

␣-Amyrin,␤-amyrina,b,d

Protiumdecandrum(Aubl.) Marchand/chico-da-silva (JFLS421)

Wounds – Resin(fat-soluble)–

flavonoid

glycosides/triterpenes

␣-Amyrin,␤-amyrin, quercetin,quercitrina,b,c,d

Protiumheptaphyllum(Aubl.) Marchand/breu-branco (JFLS458)

Headache*_{, stroke}*_, diseaseoftheair*_and respiratorydisease

Analgesic – resin (Oliveira etal.,2005a);

anxiolytic/anti-depressant– resin(Aragãoetal.,2006); anti-inflammatory–resin (Oliveiraetal.,2004)

Resin (fat-soluble) – triterpenes/polyphenolic

␣-Amyrin, ␤-amyrin, quercetin,catechina,b,c,d

Clusiaceae Calophyllumbrasiliense

Cambess./jacareúba (JFLS513)

Removewarts* _{Antiproliferative–stem}

bark (Jinetal.,2011)

Latex(fat-soluble)– phenolics

Brasiliensicacid, isobrasiliensic acida,c,d

Vismiaguianensis(Aubl.) Choisy/lacre(JFLS359)

Ringworm – Latex(fat-soluble)–

triterpenes

Lupeol,friedelin,betulinic acida,b,d

Dilleniaceae Doliocarpussp/cipód’água (JFLS423)

Strokeanddiseaseofthe air

– Latex(fat-soluble)–

triterpene

betulinicacida,b,d

Euphorbiaceae Heveaspruceana(Benth.) Müll. Arg./seringa-barriguda (JFLS401)

Fever – Latex (fat-soluble) –

triterpene

Lupeola,b,d

Fabaceae s.l.

Copaiferamultijuga

Hayne/óleo-de-copaíba (JFLS403)

Sorethroat*_{,feverandflu Anti-inflammatory–}

oleoresin(Veiga-Junior etal.,2007);antinociceptive –oil(Gomesetal.,2007)

Oleoresin(fat-soluble)– diterpene

Copalicacida,d

Hymenaeacourbaril

L./jatobá-do-mato(JFLS424)

Flu*_{, sorethroat}*_{, and} cough

Antiviral – no data found (Cecílioetal.,2012); anti-inflammatory– pericarp(Takagietal.,2002)

Resin (fat-soluble) – diterpene

Kolavenic acida

Lecythidaceae BertolletiaexcelsaHumb.& Bonpl./castanha-do-Pará (ER88)

Hemorrhoidsand diarrhea

Gum(water-soluble)– aminoacids

Lysine,tryptophana,d

Moraceae Brosimumparinarioides

Ducke/leite-do-Amapá (JFLS414)

Tuberculosis,wormand

weaknessinthelung

– Latex (fat-soluble) – fatty acids,triterpenes

palmitic acid, lupeol, ursolicacid,oleanolic acida,b,d

themostprominent.Culture-boundsyndromesarelistedasderrame (stroke)anddoenc¸adoar(airdisease),whichwillbedetailedbelow. In regard to mode of preparation used for compounding exudates-containing medicines, most partof them are used in naturawithout passing through heat process. Someexceptions are:Bertolletiaexcelsagum,usedtoprepareateabydecoction; Hymenaeacourbarilresin,usedtoprepareasyrup;and Protium resinsareusedforfumigation.Thistypeofpreparationmethodis morerelatedtoculture-boundsyndromes,whichisdiscussedbelow. Administrationroutearevaried,includingoral,topicand inhala-tion. Topic usewas observed in thecase of warts removal by Calophyllumbrasiliense.Inthiscaseitwasalsoobservedreligious context onhealing practice,since latex ofthis plant shouldbe placedonforthreefollowed Fridayswhilehealerprays;onthe lastFridaythewartfalls,accordingtowhattheyexplained and toourfield observation. LatexproducedbyVismia guianensisis alsoadministeredtopicallyonmicoses.Still,latexofPhilodendron solimoesenseis dropped directlyintotheeyes for cataract indi-cation.Oraladministration wasobserved forsomelatex,asthe onesofBrosimumparinarioidesandCoumamacrocarpa.According tointervieweesreports,theyareconsideredas‘milk’,becauseofits consistenceandwhitecolormilk.Inhalingwasobservedonlyfor resins,duringfumigationprocess,describedahead.

Usually,dosesareusedasdrops(consideringlatex,sapand ole-oresins)orfreshpieces(resinsandgums).However,dosesusedof theseexudatesonhome-brewedremedieswerenotalwaysclearly defined,andbecameeven moresubjectivewhenusedby fumi-gation,wheremanyingredientsarecarbonizedinaclaypotwith Protiumresins–leaves,animalspieces,suchasantlers,teeth,penis, birds’nests,amongmanyothers–alongwithcharcoal.Duringthis ritual,thechildmustbepassedincrossthroughsmokeproduced byfumigation,threetimesaday,whilepraying.Itisverycommon theuseofprayersduringvarioustherapeuticpracticesobserved, sincereligionisstillCatholicinsomecommunities.Thesepractices arenotobservedanymorewherereligionhasundergonechange andbecameevangelical.Thischangetriggeredthebreakupofsome communitiesunderstudy.

Basedonsolubilityandchemicalclassesobtainedduringour chemicalanalyzes,fifteenexudateswereclassifiedinto:latex(7 exudates),resins(5), sap(1), gum(1) andoleoresin(1). Eleven ofthemwerenotmentioned inpharmacologicalliteratureuntil now.Box1showsfamilyandscientificname/vernacularnameof eachplantanalyzed, classificationofexudates based onits sol-ubility,mainidentifiedcompounds,medicinaluseby“caboclos” river-dwellersand pharmacologicalstudiespublishedrelatedto thesespecificexudates.

ToidentifythemaincompoundsofeachobtainedMeOHextract, theywere individually analyzed by NMR and/or MS (LC/ESIMS orGC/EIMS)(Box 1).13_{C NMR spectraof extractsfromProtium} speciesaswellasfromHeveaspruceana,Philodendron solimoen-sis, V. guianensis, Doliocarpus sp. and B. parinarioides displayed moreintensepeaksofsp2_{carbonsatı150/109,145/122and/or} 139/124characteristicsoflupane,oleananeandursanetriterpenes skeleton(OleaandRoque,1990).Thisanalysis,inassociationwith GC/EIMSdata, afforded theidentification of ␣- and ␤-amyrins, friedelinandlupeolaswellasursolic,oleanolicandbetulinicacids. Additionally,1_{HNMRspectraofcrudeextractsfromProtium} decan-drumandProtiumheptaphyllumshowedlessintensesignalsatı

6.2–8.0,suggestingthepresenceofphenolicderivatives.13_CNMR associatedtoLC/ESIMSanalysisofcrudematerialaffordedthe iden-tificationofflavonoidsquercetin,quercitrinandcatechin.Ascrude MeOH extract from C. brasiliense showedto mainly composed, by1_{H NMR, ofaliphatic/aromatic material,it wasdirectly} ana-lyzedbyLC/ESIMS.UsingUVandMSdataaswellasco-injection usingauthenticsamples,theidentificationofbrasiliensicand iso-brasiliensicacidswaspossible.NMR(1_Hand13_{C)spectralanalysis}

of Fabaceae plants (Copaifera multijuga and H. courbaril) crude extractssuggestedthepredominanceofditerpeneacidsduethe presenceofseveralpeaksatı106–165(sp2_{carbons)andat} approx-imatelyı172(COOH).Thecomparisonofreporteddatatocopalic andkolavenicacids(Marchesinietal.,2009),allowedthe identifi-cationofthesediterpenes.The1_{HNMRspectrumofMeOHextract} fromB.excelsasuggestedtheoccurrenceofaminoacidsasmain constituents.UsingNMRdataobtainedforauthenticsamples,the identificationoflysineandtryptophanwaspossible.The identifi-cationofsteroidssitosterolandstigmasterolinthecrudeMeOH extractfromP.solimoesensewasconductedbyanalysisof1_HNMR spectrumduethecharacteristicpeaksatı0.7(s),5.4(brd)and 3.5(m).Aimingconfirmation,partofthismaterialwasdissolved inMeOH:H2O2:1andpartitionedwithhexane.Theorganicphase wasanalyzedby13_{CNMRandGC/EIMSconfirmingtheoccurrence} ofdetectedsteroidsaswellas3-octadecenylphenol.

Discussion

Basedonthesolubilityandchemicalclassesfoundineachof thefifteenexudatesanalyzed,andcomparingthemtothechemical evidencepresentedbyLangenheim(2003),weclassifiedtheminto: latex,resins,sap,gum,andoleoresin,asshowninBox1.Our classi-ficationwascorroboratedbysomeexistingstudiesfrombotanical anatomyand morphologywhichconfirmthepresenceofresins and/orresiniferouschannelsinProtiumspecies,suchasP. amazon-icum,P.aracouchini,P.decandrum,P.heptaphyllum(Rüdigeretal., 2007;Dalyetal.,2011),C.multijuga(Milanietal.,2012)andH. courbaril(Cunninghametal.,1974);andlaticifersinC.macrocarpa (Williams,1962),C.brasiliense(Cabral,2011)andB.parinarioides (Jacomassietal.,2007).

AsshowninBox1,amongthefifteenexudates,fourhavebeen describedinpreviouspharmacologicalstudiesandtheyhave pre-sentedcorrespondencebetweenatleastonelocalmedicineuseand theobservedeffectpublishedonscientificliterature.Moreover,it isnoteworthythatpharmacologicalstudiesofliteraturewere con-ductedwithplantsexudates.Anexceptionwastheantiviralcitation forH.courbarilbyCecílioetal.(2012),whichtheinvestigatedpart oftheplantwasnotmentioned.Thesefourspeciesarehighlighted below.

Itwasalsoobservedthatresinsofotherspeciesofthesame genus,P.aracouchini(Aubl.)andP.amazonicum(Cuatrec.)canalso beusedtotreatheadache,derrame(stroke)anddoenc¸adoar(disease oftheair),informofcigarettes.

Latex of ‘jacareúba’ (C. brasiliense Cambess.) is indicated to remove warts in local medicine and it has been the target of severalpharmacologicalstudies,which haveconfirmedits anti-proliferative effect onmantle cell lymphoma (Jin et al., 2011), antifungal(Reyes-Chilpaetal.,1997),anti-ulcer(Lemosetal.,2012) andtrypanocidalactivities(Reaetal.,2013).Thesebiological activ-itiescouldberelated tothepresenceof coumarins,chromenes, xanthonesandtriterpenes,sincethesecompoundswerefoundas mainconstituentsofresin(Noldinetal.,2006).

Theoleoresinof‘copaiba’(C.multijugaHayne)isusedforthe treatmentofsorethroat,feverandflu.Itisaversatilespecieamong interviewees,itsexudateisusedtoprepareasyrupwithotherplant species,andin severecasesofthroatinflammationit is recom-mendedtoputafewdropsinnaturaonaffectedsite.Insurveys conducted in Amazon region, it wasobserved similar uses for copaíba(BranchandSilva,1983;Balée,1994;PintoandMaduro, 2003;ShanleyandRosa,2005;Rodrigues,2006).Somestudieshave evaluateditsanti-inflammatoryeffects(Veiga-Junioretal.,2007), anti-oedematogenicproperties(Veiga-Junioretal.,2006), antinoci-ceptive(Gomesetal.,2007),antineoplastic(Gomesetal.,2008)and antimicrobialactivities(Santosetal.,2008).Theuseforsorethroat showedsomesimilaritywithinvestigatedeffects/actions, demon-stratingconcordancebetweenpopularknowledgeandacademic science.

The‘jatobá-do-mato’(H.courbaril)isawell-knownplantinthe AmazonaswellasinotherBrazilianbiomes,anditisfrequently citedtocurerespiratorydiseasesinethnopharmacologicalsurveys (Oliveiraetal.,2011).Amongthe“caboclos”river-dwellersliving alongtheUniniRiver,resinofthisplantisusedtotreatflu,sore throat,andcough.Cecílioetal.(2012)demonstrateditsantiviral activity,whileitsanti-inflammatoryeffectwasreportedbyTakagi etal.(2002).Thelattermayberelatedtosorethroatindication. BothstudiesutilizedEtOHextracts,fromleavesandpericarpsof theplant,respectively.Theseactivitiescouldbeassociatedtothe presenceofkolavenicacid,aclerodanetypediterpene,foundas themainderivativeoftheplantmaterial.Thisditerpenerevealed antimicrobial(Faizietal.,2008)andantifungal(Salahetal.,2003) activities.

Themajorcontributionofthisstudyliesonexudates registra-tionbyethnopharmacology,especiallythoseeleventhathavenot beeninvestigatedbypharmacologyyet.Basedonwealthofdata collectedinAmazonregion–andconsideringthefactthatvarious Brazilianbiomesalsohaveexudates-producingplants–webelieve thatagreatereffortshouldbedonebyresearchersofethnoscience areasinordertoregistertheseuses.

Conclusion

Exudatesarepromisingplantmaterialswhichcouldbeused in drug discovery due to accumulation of several metabolites. Althoughthesematerialshavebeenonly sparinglyinvestigated bypharmacologicalstudies,theresultsreportedinthisworkmay contributetochemical/pharmacologicalknowledgeoftheseplant species,manyofwhichhavebeenusedinclassicalBrazilianfolk medicine.

Ethicaldisclosures

Protectionofhumanandanimalsubjects. Theauthorsdeclare

thatnoexperimentswereperformedonhumansoranimalsfor thisstudy.

Confidentialityofdata. Theauthorsdeclarethatnopatientdata

appearinthisarticle.

Righttoprivacyandinformedconsent.Theauthorsdeclarethat

nopatientdataappearinthisarticle.

Authors’contributions

ERhasidealized,oriented,collectedsomeoftheexudates,and coordinatedthestudy.JHGLwasresponsibleforchemicalanalysis. JFLSperformedpartofthefieldworkanddatacollection.JT,PBY eFCcontributedtodiscussions,mapelaboration,review, format-tingandstandardizationofmanuscript.Allauthorshavereadfinal manuscriptandtheyhaveapprovedsubmission.

Conflictsofinterest

Theauthorsdeclarenoconflictsofinterest.

Acknowledgments

FinancialandstructuralsupportwasprovidedbyCAPES,CNPq, FAPESP/Programa BIOTA and Associac¸ão Fundo de Incentivo à Pesquisa.AuthorswouldliketothanktheinhabitantsoftheUnini River fortheir contributions tothis work.We also thank Vito-riaAmazônicaFoundation,UniniRiverExtractiveReserveandJau NationalPark,forprovidinginitialcontactswiththecommunity andlogistical support.We wouldliketoacknowledgetheplant taxonomy technician, Mr.Jose Ramos, at theNational Institute forAmazonianResearch,whoidentifiedvegetablematerials cor-respondingtoexudates.Wealsothanktwoanonymousreferees fortheirhelpfulsuggestionsinimprovingthispaper.

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