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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.
SCREENING OF PLANTS FOUND IN THE STATE OF AMAZONAS, BRAZILFOR LARVICIDAL ACTIVITY AGAINST Aedes aegypti LARVAE
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.
SCREENING OF PLANTS FOUND IN THE STATE OF AMAZONAS, BRAZILFOR LARVICIDAL ACTIVITY AGAINST Aedes aegypti LARVAE
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.
SCREENING OF PLANTS FOUND IN THE STATE OF AMAZONAS, BRAZILFOR LARVICIDAL ACTIVITY AGAINST Aedes aegypti LARVAE
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.
SCREENING OF PLANTS FOUND IN THE STATE OF AMAZONAS, BRAZILFOR LARVICIDAL ACTIVITY AGAINST Aedes aegypti LARVAE
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.
SCREENING OF PLANTS FOUND IN THE STATE OF AMAZONAS, BRAZILFOR LARVICIDAL ACTIVITY AGAINST Aedes aegypti LARVAE
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.
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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.)
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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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RECEBIDO EM 30/12/2002ACEITO EM 07/01/2004