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97 VOL. 34(1) 2004: 97 - 105

Screening of plants found in the State of Amazonas,Brazil for larvicidal activity against Aedes aegypti larvae.

Adrian Martin POHLIT,a,1 Etienne Louis Jacques QUIGNARD,a Sergio Massayoshi NUNOMURA,a WanderliPedro TADEI,b Ari de Freitas HIDALGO,c Ana Cristina da Silva PINTO,d Elba Vieira Mustafa dos SANTOS,d

Sabrina Kelly Reis de MORAIS,d Rita De Cássia Guedes SARAIVA,e Lin Chau MING,f Alexandre MascarenhasALECRIM,a Alfeu de Barros FERRAZ,a Andreza Cristiana da Silva PEDROSO,a Elisangela Vieira DINIZ,a EllenKathryn FINNEY,a Erika de Oliveira GOMES,a Hercules Bezerra DIAS,a Katiuscia dos Santos de SOUZA,a LauraCristina Pereira de OLIVEIRA,a Luciana de Castro DON,a Maria Mireide Andrade QUEIROZ,a MarycleumaCampos HENRIQUE,a Mirian dos SANTOS,a Orivaldo da Silva LACERDA JÚNIOR,a Patrícia de Souza PINTO,a

Suniá Gomes SILVA.a Yara Rodrigues GRAÇA.a

ABSTRACTEthanol, methanol and water extracts representing mostly native plant species found in the Amazon region were prepared,respectively, by maceration, continuous liquid-solid extraction and infusion, followed by evaporation and freeze-drying.The freeze-dried extracts were tested for lethality toward Aedes aegypti larvae at test concentrations of 500 µg / mL. Ingeneral, methanol extracts exhibited the greatest larvicidal activity. The following 7 methanol extracts of (the parts of) theindicated plant species were the most active, resulting in 100 % mortality in A. aegypti larvae: Tapura amazonica Poepp.(root), Piper aduncum L. (leaf and root), P. tuberculatum Jacq. (leaf, fruit and branch). and Simaba polyphylla (Cavalcante)W.W. Thomas (branch).

KEY WORDSlarvicide, Aedes aegypti, Tapura, Piper, Simaba, Amazonia.

Triagem de plantas encontradas no Estado do Amazonas paraatividade larvicida contra Aedes aegypti.

RESUMOExtratos aquosos, etanólicos e metanólicos, representando principalmente espécies vegetais nativas encontradas naregião Amazônica, foram preparados, respectivamente, por infusão, maceração e extração contínua líquido-sólido,seguida de evaporação e liofilização. Os extratos liofilizados foram testados para atividade contra larvas de Aedesaegypti, na concentração única de 500 µg / mL. Os extratos metanólicos foram, em geral, os que apresentaram maioratividade larvicida. Os seguintes 7 extratos metanólicos das (partes das) espécies vegetais indicadas foram os maisativos, provocando 100 % de mortalidade em larvas de A. aegypti: Tapura amazonica Poepp. (raiz), Piper aduncum L.(folha e raiz), P. tuberculatum Jacq. (folha, fruto e galho) e Simaba polyphylla (Cavalcante) W.W. Thomas (galho).

PALAVRAS-CHAVElarvicida, Aedes aegypti, Tapura, Piper, Simaba, Amazônia.

1 Author to whom correspondence should be addressed (e-mail: [email protected]).a Coordenação de Pesquisas em Produtos Naturais (CPPN), b Coordenação de Pesquisas em Ciências da Saúde (CPCS), Instituto Nacional de Pesquisas

da Amazônia (INPA), Av. André Araújo, 2936, 69083-000 Manaus, Amazonas; c Faculdade de Ciências Agrárias (FCA), d Curso de Pós-graduação emQuímica de Produtos Naturais, Universidade Federal do Amazonas (UFAM), Av. Gal. Rodrigo O. J. Ramos, 3000, 69077-000 Manaus, Amazonas; e Cursode Pós-graduação em Química, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Rio de Janeiro; f Universidade do Estado de São Paulo(UNESP), Fazenda Experimental Lageado, 237, 18603-970 Botucatu, São Paulo.

98 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.

SCREENING OF PLANTS FOUND IN THE STATE OF AMAZONAS, BRAZILFOR LARVICIDAL ACTIVITY AGAINST Aedes aegypti LARVAE

INTRODUCTION

In response to strong selective pressures of herbivorousinsects, toxic secondary metabolites have evolved in plantsand often affect insect nerve function and behaviour(Sharma et al., 1998). Plant extracts show potential assources of natural larvicides for the control of mosquitovectors and an early review has been published in whichmore than 300 larvicidal plant species were identified(Sukamar et al., 1991). In recent times, studies on the activityof plant extracts towards Aedes sp. larvae from different partsof the world, such as, North America (Bergeron et al., 1996),Argentina, Bolivia, Brasil and Peru (Chantraine et al., 1998;Ciccia et al., 2000; Macedo et al., 1997), Trinidade andTobago (Chariandy et al., 1999), Mali (Diallo et al., 2001),Negev Desert (Sathiyamoorthy et al., 1997) and Africa(Marston et al., 1993), among others, have been publishedand have revealed numerous examples of active plantextracts representing diverse taxonomic groups. Moresystematic and directed studies have revealed a number ofvery active plant extracts, essential oils, and isolatedlarvicidal phytochemicals (Bandara et al., 2000; Bernard etal., 1995; Hostettmann & Potterat, 1997; Latha & Ammini,2000; Lee, 2000; Oberlies et al., 1998; Park et al., 2002;Pushpalatha & Muthukrishnan, 1999; Rahuman et al., 2000;Sharma et al., 1998; Thorsell et al., 1998). In the presentstudy, we have screened polar extracts of mostly native plantsfrom the Brazilian Amazon for activity against larvae of thehemorrhagic dengue fever vector, Aedes aegypti (Diptera:Culicidae), at a single test concentration of 500 µg / mL.Data (species names, plant parts, extraction solvent) for bothactive and inactive extracts are presented.

MATERIAL AND METHODS

Plant collecting, processing,and extract preparation.

Plants were collected from 1999 to 2002 in the Stateof Amazonas. Some genera (for example, Piperand Aspidosperma) and families (Euphorbiaceae,Simaroubaceae, Apocynaceae, Piperaceae, etc.) were betterrepresented among the plants collected due to our specificinterest in the bioactivity of these taxonomic groups. Theplant materials were generally dried on the ground, in theshade, then stored in plastic bags under the protectivewarmth and luminosity of incandescent lighting. Wherepossible, fertile specimens of plant species were collected,used for identification and deposited at the INPAHerbarium. Many samples for study were collected fromindividual trees in INPA’s Ducke Reserve that had beencatalogued and identified in recent systematic studies(Ribeiro et al., 1999). Ethanol, methanol and water extractswere prepared, respectively, by maceration, continuousliquid-solid extraction, and infusion, followed by rotaryevaporation under vacuum with heating in a bath to ca. 40-45 ºC and freeze-drying. After this last step, extracts were

presumed to be solvent-free. Dry extracts were depositedin our extract bank at the Coordenação de Pesquisas emProdutos Naturais (CPPN / INPA) where they were storedat –19 ºC. Many of these extracts were screened previouslyby some of us against larvae of the brine shrimp speciesArtemia franciscana (Quignard et al., 2003).

Assay for larvicidal activityagainst Aedes aegypti.

Aedes aegypti larvae were obtained by incubation ofeggs from a laboratory colony at the Coordenação dePesquisas em Ciências da Saúde (CPCS/INPA) in tap water.Stock solutions or suspensions of extracts in DMSO or de-ionized water (50 mg / mL) were prepared. Each stocksolution (100 µL) was then transferred to plastic cupscontaining 10 third instar (three-day old) larvae in tapwater having a final volume of 10 mL and a final extractconcentration of 500 µg / mL. Negative controls wereprepared analogously, substituting 100 µL of tap water orDMSO for sample solution in each cup, resulting in a finalDMSO concentration of 1 %. Each experiment was run intriplicate and compared with a control set after 24h at26 – 27 ºC. Dead larvae were counted and the larvicidalactivity expressed as % mortality based on live larvaepresent initially. In general, no dead larvae were observedin the controls after 24 h.

RESULTS AND DISCUSSION

Table 1 presents ethnobotanical and other informationon the species investigated in this study. A number of theseplants are traditionally used medicinal plants for thetreatment of malaria, fevers, liver problems, and othersymptoms caused by malaria infections, whose study is amajor focus of our research group. Other plants studiedwere collected using a random sampling technique in INPAforest reserves in or near Manaus (Reserva Florestal AdolphoDucke, Reserva da Campina / ZF-2). Still other species, fromsuch genera as Aspidosperma (Apocynaceae) and Piper(Piperaceae), or from families such as Simaroubaceae, werestudied here as part of a systematic approach to theinvestigation of the chemistry and biological activity ofAmazon flora of these, and other taxonomic groups. Thus,the plants studied herein were not selected a priori fortheir known insecticidal, larvicidal or repellent properties.The discovery of larvicidal activity, as described below, servesas a demonstration of the power of bioprospection in thediscovery of new larivicidal extracts and compounds basedon many plants for which no such activity has beendescribed or is known in the literature.

Tables 2 and 3 present, respectively, data for larvicidaland inactive plant extracts. Comparison of the resultspresented in Tables 2 and 3 reveals in a general way thatextracts prepared from the less polar solvents ethanol andmethanol were more active against A. aegypti larvae than

99 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.


Tabl

e 1

- Eth

nobo

tani

c an

d ot

her

info

rmat

ion

for

the

plan

ts s

tudi

ed.

cont. >

YLIM

AFe

mancifitneicS

eman

nom

moC

traP)sei-(ytreporp

/)s(esu

rehtodnalanicide

MfeR

EAEC

AIDRAC

AN

Anib

mom

saidnopS.L

ájac,áberepat

,tiurf,fael

krab,toor

lanigav;aehrraid

,revef,airala

mfo

tnemtaert

snoitcefni2

,1

EAEC

ANYC

OPA

agnacararaa

mrepsodipsA

arierreF-sednacraM

eert4

muhtnamsed

.A

htneB.

.grA.llü

Mxe

agnacararafael

reveffo

tnemtaert

6,4

munaivargcram

.A

nosdooW

abúanaparackrab

,doow

noitcurtsnoc;recnac

,setebaid,airala

mfo

tnemtaert

4,

1muditin

.A

.grA.llü

Mxe

.htneBabúanaparac

xetal,krab

,doow

noitcurtsnoc;recnac

,ysorpel,airala

mfo

tnemtaert

4,1

munaihtiwdnas

.A

.rgkraM

eert4

iisetluhcs.

Anosdoo

Wkrab

;sesocym

,reveffo

tnemtaert

setimret

tsniaganoitcetorp

7,4

munaecurps.

A.grA

.M

xe.htneB

doow

noitcurtsnoc4

,3iisagrav

.A

.C

D.A

krabsetib

tcesni,sdnuo

w;revef

fotne

mtaert7

,2muetnegra

mumrepsossie

Gnosdoo

Wacnarb

arauqiracakrab

airalam

;reveffo

tnemtaert

4,2

mutaloecru.

Gyrtne

G.

H.Aacnarb

arauqiracakrab

4abuucus

suhtnatami

Hnosdoo

W).grA

.llüM

xeecurpS(

abúucusxetal

,serutcarfenob

,reveffo

tnemtaert

sehcahtoot4

,1

EAEC

ARETSA

asolipsnediB

.Loterp-oãcip

tnalpeloh

w,yspord

,sititapehfo

tnemtaert

citeruid;yretnesyd

5,1

allemca

sehtnalip Syarru

M).L(

úbmaj

rewolf

sisolucrebut,sesaesid

gnulfo

tnemtaert

1E

AECAI

NO

NGIB

anacniaiubebaT

yrtneG

.H.A

ocra'duap

krabsro

mut;sisaididnac

fotne

mtaert1

EAEC

AXIBanallero

axiB.L

mucurufael

,toohs;sesaesidlaerenev

,reveffo

tnemtaert

caisidorhpa,citpesitna

1

EAEC

ARESRUB

inihcuocaramuitorP

dnahcraM

).lbuA(ohle

mrevuerb

niserevitaxal

9E

AECAIS

ULCesneilisarb

mullyhpolaC

.ssebma

Cabúeracaj

krabmsita

muehr,s

mrow

,setebaid,aehrraid

fotne

mtaert1

EAEC

ATIBRUC

UCaitnarahc

acidromo

M.L

onateaC

oãSed

oãlem

tnalpeloh

wcite

me;s

mrow

,spmarclanitsetni

fotne

mtaert1

100 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.


Tabl

e 1

- Eth

nobo

tani

c an

d ot

her

info

rmat

ion

for

the

plan

ts s

tudi

ed. (

cont

.)

cont. >

EAE

CA

REPY

Csisnetarp

airelcSsee

Nxe

.ldniLaciririt

toor,fael

smelborp

cirtsag,sp

marclanitsetnifo

tnemtaert

01,6E

AEC

ALATEP

AH

CID

acinozama

arupaT.ppeoP

arupatfael

cixot7

EAE

CAI

BR

OHP

UEaracujac

notorC

.htneBacacas

krabaehrraid

;setebaid,noita

mmalfni

revilfo

tnemtaert

1sisne

wuojnal.

C.lbaJ

amid

krabrevef

fotne

mtaert01

,2nolyxorelcs

sispordnarciM

seugirdoR.A.

Wírup

aca,ohnizoãip

eertE

AEC

AN

AIT

NEG

arolfidnargaihcaT

eriugaM

&revae

Weertlla

ms4

EAE

CAI

MAL

muhtnarcim

mumic

O.dlli

Wacavafla

faelairala

mfo

tnemtaert

5,2

EAE

CA

RU

ALarodoeasor

abinA

ekcuD

asoruap

liosesu

cigolotnodo;

msitamuehr

,seigrellafo

tnemtaert

5E

AEC

ADI

HT

YCEL

atlaarohpohtyro

ChtunK

.R)ohle

mrev(oriepir

eert4

EAE

DIOI

NIPLASE

AC

:EAS

ONI

MU

GELasoutsaf

aissaC

legoVxe

.dlliW

oruo-ed-avuhceert

tnemanro

11anaecurps

.C

.htneB-ad-ira

m-iram

emrif-arret

toorrevef

fotne

mtaert4

,2

silatnediccoanneS

kniL).L(

,otsapata

maboirejna

mdees

,toor,airala

m,revef

fotne

mtaertcixot

,evitroba;sitihcnorb

,amhtsa

2,1

EAE

DIO

NOILIP

AP:E

ASO

NIM

UGEL

sisnecauorpaiztra

wSffohs

mA).lbuA(

itutum

eert8

EAE

CA

MREPSI

NEM

ailofidnargatub

Ahti

wdnaS).tra

M(atob

edópic

fael,

mets,toor

ytilirets,ai

mena,airala

mfo

tnemtaert

1E

AEC

AC

ALO

sisnenaiugaitrauqni

M.lbuA

ahlemrev

arauqiracakrab

,doow

nosiophsif

;noitcurtsnoclivic,stsop

1E

AECAREPIP

mucnudarepiP

.Lagnol

atnemip

tiurf,fael

citocymitna

,laiborcimitna

,citpyts,cita

mora7

.Pesneapa

ma.kcnuY

elepi-ipatmets

,faelssenizzid

fotne

mtaert9

snaccab.P

.C

D.

C).qi

M(burhs

munairatipac.P

.kcnuYburhs

mudopotryc.P

.C

D.

Cmi

macajfael

shtabcita

mora01

mutatalid.P

.hciRburhs

seitreporplagnufitnamullipitcere

.P.kcnuY

burhs

101 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.


pmunainna

mtsoh.P

.C

D.

C).qi

M(ollicnodroc

faelstra

wfo

tnemtaert

7mutalucrebut

.P.qcaJ

agnolatne

mipfael

nosiophsif

7,6atatlep

ehpromohtoP

.qiM

).L(abepaac

faelcite

me,citeruid

;sreveffo

tnemtaert

1E

AECAC

ALUTR

OPasolip

acalutroP.L

açnoed

rolftnalp

elohw

.citeruid;s

mrow

,sitirhpen,sititapeh

fotne

mtaert1

EAEC

AN

MA

HRsucinoza

masuhpyzizolep

mA

ekcuD

árim-arucaras

fael,krab

,toorsetib

tcesni,airala

mfo

tnemtaert

2,1

EAEC

ARO

HPOZI

HRsisnenaiug

aeruopissaC

.lbuAovarb

áçara4

EAEC

AIBUR

atadnuaigrebnedaL

hcsztolKaniuq

aniuqkrab

,faelairala

m,revef

fotne

mtaert8

EAEC

ATUR

ailofitnaruasurti

Celgni

wS).tsirh

C(ogib

mued

ariemil

toorairala

mfo

tnemtaert

2E

AECAT

OPAS

asolunevsilohporci

MerreiP

)relhciE&

.traM(

acnarbanaruiba

tiurfdoof

1E

AECAB

UOR

AMIS

iecurpsa

mmelorciP

.f.koo

Hanarefac

sitirtsag,revef

,airalam

fotne

mtaert4,2

allyhpylopaba

miSsa

mohT.W.

W)etnaclava

C(eert

4aba

miS.von

.pseert

4ara

maabuora

miS.lbuA

ápuram

krab,aispepsyd

,aimena

,reveffo

tnemtaert

4,1E

AECA

NAL

OSatalugna

silasyhP.L

acallum

aslobúpa

mactiurf

,fael,a

mhtsa,airala

mfo

tnemtaert

citocran;noita

mmalfni

2,1

Tabl

e 1

- Eth

nobo

tani

c an

d ot

her

info

rmat

ion

for

the

plan

ts s

tudi

ed. (

cont

.)

Ref

eren

ces

(ref

) cite

d: 1

Duk

e &

Vas

quez

(199

4), 2

Mill

iken

(199

7), 3

Lor

enzi

(199

8), 4

Rib

eiro

et a

l. (1

999)

, 5 R

evill

a (2

002)

,6

Mor

s et

al.

(200

0), 7

Sch

ulte

s &

Raf

fauf

(199

0), 8

Pio

Cor

rêa

(197

8), 9

Gre

nand

et a

l. (1

987)

,10

Silv

a et

al.

(197

7), 1

1 Le

Coi

nte

(194

7).

102 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.


Table 2 - Data for Plant Extracts Exhibiting Lethality to Aedes aegypti Larvae.

cont. >

YLIMAF traP 1 tcartxE 2 ytilatroMemancifitneicS )%(EAECANYCOPA

agnacararaamrepsodipsA arierreF-sednacraM kB M 4munaivargcram.A nosdooW kB W 4

iisetluhcs.A nosdooW kB M 33EAECARETSA

B asolipsnedi .L tR M 01allemcasehtnalipS yarruM).L( pW M 73

EAECANGONGIBaiubebaT anacni yrtneG.H.A kB E 4

EAECAISULCesneilisarbmullyhpolaC .ssebmaC kB W 02

EAECATIBRUCUCaitnarahcacidomroM .L tS M 7

EAECALATEPAHCIDacinozamaarupaT .ppeoP rB M 7

tR M 001EAECAIBROHPUE

aracujacnotorC .htneB fL M 4rB M 38

sisnewuojnal.C .lbaJ kB,rF M 09dS M 03

nolyxorelcssispordnarciM seugirdoR.A.W tR M 7EAECANAITNEGarolfidnargaihcaT revaeW&eriugaM tS M 7

fL M 01rB M 03tR E 46

EAECARUALarodoeasorabinA ekcuD fL M 34

rB W 33dW M 32

EAECAREPIPmucnudarepiP .L fL M 001

tR M 001esnepama.P .kcnuY fL M 71

tS,rB W 4munairatipac.P .kcnuY fL M 4

mutatalid.P .hciR tS,rB M 7munainamtsoh.P .CD.C).qiM( fL M 71

tS,rB M 7mutalucrebut.P .qcaJ fL M 001

rF M 001rF W 35tS,rB M 001

atatlepehpromohtoP .qiM).L( fL E 7tR E 7

103 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.


Table 3 - Inactive Plant Extracts.

cont. >

EAECABUORAMISiecurpsammelorciP .kooH tS M 75

tR M 47W 44

allyhpylopabamiS samohT.W.W)etnaclavaC( rB M 001S abami .von.ps rB M 07

aramaabuoramiS .lbuA rB M 73

1 - Bk – bark; Br – branch; Fl – flower; Fr – fruit; Lf – leaf; Rt – root; Sd – seed; St – stem; Wd – wood; Wp – whole plant.2 - E – ethanol; M – methanol; W – water.

Table 2 - Data for Plant Extracts Exhibiting Lethality to Aedes aegypti Larvae. (cont.)

ocifítneiCemoN )s(traP 1 ])s(tcartxE[ 2

arolfidnargatubA htiwdnaS).traM( ]W,M[VsucinozamasuhpyzizolepmA ekcuD ]W,M[V

arodoeasorabinA ekcuD ]W[dW,]W[fL,]M[rBagnacararaamrepsodipsA arierreF-sednacraM ]W[kB

muhtnamsed.A .grA.llüMxe.htneB ]W,M[kBmunaivargcram.A nosdooW ]M[kB

muditin.A .grA.llüMxe.htneB ]W,M[kBmunaihtiwdnas.A .rgkraM ]W,M[kB

iisetluhcs.A nosdooW ]W[kBmunaecurps.A .grA.Mxe.htneB ]M[kB

iisagrav.A .CD.A ]M[kBasolipsnediB .L ]W[tRanalleroaxiB .L ]W,M[fL

esneilisarbmullyhpolaC .ssebmaC ]W[kBasoutsafaissaC legoVxe.dlliW ]W[tR,]M[fL

anaecurps.C .htneB ]E[tRsisnenaiugaeruopissaC .lbuA ]M[tR

elgniwS).tsirhC(ailofitnaruasurtiC ]W[TRatlaarohpohtyroC htunK.R ]M[tR

aracujacnotorC .htneB ]W,M[fL,]W[rBsisnewuojnal.C .lbaJ ]W[dS,]W,M[fL,]W[rF,]W,M[rB

muetnegramumrepsossieG nosdooW ]M,E[kBmutaloecru.G yrtneG.H.A ]M,E[kB

abuucussuhtnatamiH nosdooW).grA.llüMxeecurpS( ]W,M[kBatadnuaigrebnedaL hcsztolK ]W,M[kBasolunevsilohporciM erreiP)relhciE&.traM( ]M[tRsisnenaiugaitrauqniM .lbuA ]M[kB

aitnarahcacidromoM .L ]W[fL/tSmuhtnarcimmumicO .dlliW ]M[fL/lF

atalugnasilasyhP .L ]W,M[fL/tS,]W,M[tS,]W,M[tRiecurpsammelorciP kooH ]W[tS,]W,M[fL

mucnudarepiP .L ]W,M[tS,]W[tR,]W[fL,]W,M[rBesneapama.P .kcnuY ]M[tR,]W,M[fL,]M[rB

snaccab.P .CD.C).qiM( ]W[tR,]W,M[fL,]W,M[rB

104 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.


the extracts prepared by extraction with the more polarwater as solvent, which is probably an indication that toxicityto these larvae should be associated in general withsubstances of medium or even low polarity.

Our results confirmed the toxicity of several plantspecies which have been described as being toxic or areused traditionally for their toxicity. For example,methanol root extract of Tapura amazonica Poepp. washighly lethal to A. aegypti larvae which is interesting inlight of the fact that it has been described by Colombiannative indians as being a toxic species (Table 1; Schultes& Raffauf, 1990). In similar fashion, P. tuberculatum Jacq.presented highly lethal extracts in the assay and is,according to traditional use, an effective fish poison(Table 1). Lastly, Picrolemma sprucei Hook (caferana),which exhibited larvicidal root and stem extracts in thepresent study, is known for its toxic effects towards thehuman fetus, being used widely in the State of Amazonasand Manaus as an abortifacient tea (Pohlit et al.,unpublished data).

Another Piper species, P. aduncum L., exhibited goodactivity against Aedes aegypti (Table 2) and is known toproduce the larvicidal phytochemical compound,dillapiole, which has been shown by Bernard et al.(1995) to be active against another Aedes sp.. Given thatP. aduncum is a fast-growing, invasive species, it ispotentially useful as an industrial source of dillapioleand other active phytochemicals.

Another interesting observation is that larvicidalactivity is definitely specific to certain parts of plants,such as P. aduncum, which exhibited very active methanolleaf and root extracts (Table 2), however, extracts of otherparts of this plant were inactive (Table 3). Other examplesof this specificity of insecticidal activity to certain plantparts is evidenced by comparing the data presented inTables 2 and 3.

Finally, it is important to note that even the lowmortality exhibited by some extracts during screening canbe significant for the discovery of bioactive phytochemicals.An example of this is the weak activity of the ethanol rootand leaf extracts of Pothomorphe peltata (L.) Miq. (eachexhibiting only 7 % mortality at 500 µg / mL, Table 2). Infurther work, bioassay-guided fractionation of this ethanolroot extract in our labs yielded very active intermediatefractions and isolation of the phytochemical larvicide, 4-nerolidylcatechol, which exhibited very significant lethalitytowards Aedes aegypti larvae and was shown bychromatographic analytical methods to be a majorcomponent of both root and leaf extracts (Pinto, 2002).

CONCLUSION

A number of larvicidal plants were identified during thiswork. Several of these are now under biomonitoredphytochemical analysis and should yield further examplesof isolated larvicidal phytochemicals in the future.

mudopotryc.P .CD.C ]W[tR,]W,M[fL,]W,M[rBmutatalid.P .hciR ]W,M[tR,]W,M[fL,]W[rBmulipitcere.P .kcnuY ]W,M[tR,]W,M[fL,]W,M[rB

munainnamtsoh.P .CD.C).qiM( ]W[fL,]W[rBmutalucrebut.P .qcaJ ]W[fL,]W,M[rF,]W[rBasolipacalutroP .L ]W,M[fL/tS

atatlepehpromohtoP .qiM).L( ]W,M[tRinihcuocaramuitorP dnahcraM).lbuA( ]W,M[tR

sisnetarpairelcS seeNxe.ldniL ]W,M[tR

silatnediccoanneS kniL).L( ]W[fL,]W[rB,]W[tR

allyhpylopabamiS samohT.W.W)etnaclavaC( ]W,E[rB,]W,M[fL,]W,E[rBabamiS .von.ps ]W,M[fL,]W[rB

aramaabuoramiS .lbuA ]W,M[fL,]W[rBallemcasehtnalipS yarruM).L( ]W[pW

nibmomsaidnopS .L ]M[kBsisecauorpaiztrawS ffohsmA).lbuA( ]W[fL,]W[rB

anacniaiubebaT yrtneG.H.A ]M[kBarolfidnargaihcaT revaeW&eriugaM ]E[tS,]W,M[tR,]W,M,E[fL,]W,E[rBacinôzamaarupaT .ppeoP ]W[tR,]M[kB,]M[fL

1 - Bk – bark; Br – branch; Fl – flower; Fr – fruit; Lf – leaf; Rt – root; Sd – seed; St – stem; Wd – wood; Wp – whole plant.2 - E – ethanol; M – methanol; W – water.

Table 3 - Inactive Plant Extracts. (cont.)

105 VOL. 34(1) 2004: 97 - 105 • POHLIT et. al.


ACKNOWLEDGEMENTS

The authors proudly recognize the financial support providedby the Brazilian National Council for Scientific and TechnologicalDevelopment (CNPq; Grant nos. 520.354/99-0 and 550.260/01-3). The following people are thankful to these institutions forscholarships: ELJQ (DTI-MCT), SMN (DCR-CNPq), ACSP and EVMS(Mestr-CAPES), SKRM (AP-CNPq), MCH (IC-CNPq), KSS (AT-CNPq),AMA, ACSP, LCD, EOG, PSP and SGS (PIBIC-INPA-CNPq).

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