Larvicidal activities against Aedes aegypti of some Brazilian medicinal plants

Bioresource Technology 98 (2007) 2549–2556

Larvicidal activities against Aedes aegypti of someBrazilian medicinal plants

M.C. de Omena a, D.M.A.F. Navarro b, J.E. de Paula c, J.S. Luna a,M.R. Ferreira de Lima a, A.E.G. Sant’Ana a,*

a Instituto de Quımica e Biotecnologia, Universidade Federal de Alagoas, 57.072-970 Maceio, AL, Brazilb Departamento de Quımica Fundamental, Universidade Federal de Pernambuco, 50.970-901 Recife, PE, Brazil

c Departamento de Botanica, Universidade de Brasilia, 70.910-900 Brasılia, DF, Brazil

Received 11 August 2004; received in revised form 7 September 2006; accepted 11 September 2006Available online 28 November 2006

Abstract

Larvicidal activities against Aedes aegypti have been determined in the ethanolic extracts obtained from 51 Brazilian medicinal plants.Eleven of the 84 extracts studied showed significant (LC50 < 100 lg mL�1) activities against larvae, with extracts from Annona crassiflora

(root bark, LC50 = 0.71 lg mL�1; root wood, LC50 = 8.94 lg mL�1) and Annona glabra (seed, LC50 = 0.06 lg mL�1) showing the high-est activities. The results obtained should be of value in the search for new natural larvicidal compounds.� 2006 Elsevier Ltd. All rights reserved.

Keywords: Larvicidal activities; Mosquito control; Aedes aegypti

1. Introduction

The extensive and diverse flora of Brazil is a naturalasset of immense potential value for the provision of sec-ondary compounds, many of which offer potential appli-cation in the manufacture of food additives, cosmetics,agrochemicals and, especially, pharmaceuticals. In SouthAmerica, the use of medicinal plants contributes signifi-cantly to primary health care, while in Brazil many plantsare used in the form of crude extracts, infusions or plastersto treat common infections. The common therapeuticproperties of plant extracts involving, for example, anti-fungal, anti-bacterial, anti-parasitic, anti-neoplastic andimmunomodulator activities have been well exploited, butthe search for new compounds with larvicidal and insecti-cidal activities is becoming more important as the region

0960-8524/$ - see front matter � 2006 Elsevier Ltd. All rights reserved.

doi:10.1016/j.biortech.2006.09.040

* Corresponding author. Tel.: +55 82 3214 1388; fax: +55 82 3214 1389.E-mail addresses: [email protected], antonio.euzebio@pesquisador.

cnpq.br (A.E.G. Sant’Ana).

is currently experiencing a dramatic increase in the inci-dence of insect-transmitted diseases.

Mosquitoes are responsible for the spread of more dis-eases than any other group of arthropods. Of particularinterest is Aedes aegypti because of its role as a vector forthe arboviruses responsible for yellow fever and denguefever, both of which are endemic to Central and SouthAmerica, Asia and Africa (Consoli and Oliveira, 1994;Neves, 2000). Dengue viruses occur as four antigenicallyrelated but distinct serotypes, which cause a broad rangeof disease, including clinically asymptomatic forms, classicdengue fever (characterised by the sudden onset of fever,headache, retro-orbital pain and myalgia), and the moresevere forms such as dengue hemorrhagic fever-dengueshock syndrome (Fundacao Nacional de Saude, 2002).Dengue fever was first described during an epidemic inPhiladelphia in 1780, and intermittent pandemics haveaffected Asia, Africa and the Americas at intervals of 10–30 years. After World War II, marked changes in humanand vector ecology fostered the transmission of multipledengue serotypes, and the tropical world is currently in

mailto:[email protected]

mailto:antonio.euzebio@pesquisador. cnpq.br

mailto:antonio.euzebio@pesquisador. cnpq.br

2550 M.C. de Omena et al. / Bioresource Technology 98 (2007) 2549–2556

the grip of a continuing dengue pandemic (Teixeira et al.,1999). Dengue is present in more than one hundred coun-tries and threatens the health of approximately 2.5 billionpeople. Worldwide, around 80 million people are infectedannually at an attack rate of 4% (Monath, 1994). In Brazil,the incidence of classical and hemorrhagic dengue isaround 500,000 cases annually and, with an index ofmortality of 5%, the disease causes 24,000 deaths per year(Teixeira et al., 1986, 1999; Fundacao Nacional de Saude,1996).

An obvious method for the control of mosquito-bornediseases is the use of insecticides, and many syntheticagents have been developed and employed in the field withconsiderable success. However, one major drawback withthe use of chemical insecticides is that they are non-selec-tive and could be harmful to other organisms in the envi-ronment. The toxicity problem, together with the growingincidence of insect resistance, has called attention to theneed for novel insecticides (Macedo et al., 1997), and formore detailed studies of naturally-occurring insecticides(Ansari et al., 2000).

Pyrethrum, extracted from Chrysanthemum cin-nerariaefolium (Asteraceae), is one of the best known ofthe natural insecticides and has provided a lead for the syn-thesis of various pyrethroid derivatives. Following the dis-covery of this phytotoxin, numerous studies have beenconducted concerning the larvicidal properties of plantsin general (Heal and Rogers, 1950; Evans and Raj, 1988;Green et al., 1991; Chantraine et al., 1998; Loset et al.,2000; Ciccia et al., 2000; Vahitha et al., 2002). Not surpris-ingly, the family Asteraceae has attracted much researchinterest (Wang et al., 1990; Macedo et al., 1997) particu-larly with respect to polyacetylene compounds with stronglarvicidal activity (Arnason et al., 1981). Compoundsderived from plants of other families, particularly theAnnonaceae, have, however, exhibited significant activitiesagainst, for example, Culex larvae (Moeschler et al., 1987;Mikolajczak et al., 1988, 1989; Mehra and Hiradhar, 2000;George and Vincent, 2005) and further screening is clearlywarranted.

Based on ethnobotanical, pharmacological and chemo-systematic criteria, we prepared 84 ethanolic extracts fromvarious tissues of 51 species of plants, belonging to 23 fam-ilies. We have screened these extracts for larvicidal activityagainst the fourth instar (L4) of A. aegypti using the well-tested methodology described by the World Health Organi-sation (1981).

2. Methods

2.1. Plant material

Fresh material (2000 g) of each of the selected plant spe-cies (see Table 1) were collected and specimens classifiedsystematically by Prof. Jose Elias de Paula (Universidadede Brasilia, DF, Brasil) and Rosangela P. de Lira Lemos(Instituto do Meio Ambiente do Estado de Alagoas-

IMA, Maceio AL, Brasil). Voucher specimens of all ofthe plant species studied were deposited in the herbariaof the respective institutes.

2.2. Preparation of extracts

Selected parts of freshly collected plant material wereseparated (if required) and immediately air dried. Driedmaterial (at least 500 g) was ground in a Nogueira (Itapira,SP, Brasil) laboratory mill to a moderately fine powder(mesh size 2.5 mm), that was extracted three times, eachwith 1.5 L of 95% ethanol, in a percolator for 3 days, orin a Soxhlet apparatus for 2 days, and the extract evapo-rated under reduced pressure in a rotary evaporator. Ifwater remained in the concentrated extract, the materialwas either freeze-dried or stored in a vacuum desiccatorover silica gel. The resulting crude extracts were kept in afreezer at �20 �C until required for assay. A stock solution(concentration 1000 lg mL�1) of each extract was preparedin dechlorinated tap water containing 1% dimethylsulphox-ide (DMSO: Aldrich, Milwaukee, WI, USA) by sonicationin a Cole Parmer (Vernon Hills, IL, USA) model 8892E-MT ultrasound bath (120 W; 47 kHz) for 20 min.

2.3. Bioassays

Eggs of A. aegypti were hatched by submerging them indechlorinated tap water at a temperature in the range 25–27 �C. The dechlorinated tap water was obtained by filtra-tion through a charcoal filter followed bubbling air for48 h. For the tests, each of which was replicated four times,ten L4 larvae of A. aegypti, in an appropriate volume ofdechlorinated water, were placed in a container and thefinal volume adjusted to 100 mL with the amount of extractstock solution required to yield a final concentration of200 lg mL�1 for a preliminary test. In order to establishLC10, LC50 and LC90 values (i.e. the concentrations ofextracts in ppm required to kill 10%, 50% and 90% of lar-vae, respectively, within 48 h), multiple 10-fold dilutions ofthe extract stock solution were prepared to provide a work-ing concentration range. Four replicate assays were carriedout for every sample concentration in a final volume of100 mL, each with 25 L4 larvae.

All assays were conducted in darkness in the range 25–27 �C and at a constant pH of 5.0. During each assay, food(proprietary cat food) was offered to the larvae. Larvaewere observed (briefly under laboratory light) at the startof the assay (t0) and at 24 and 48 h, and were considereddead when they did not respond to stimulus or when theydid not rise to the surface of the solution (World HealthOrganisation, 1981). Negative controls accompanied eachassay and involved treating larvae with dechlorinated wateror with 1% DMSO in dechlorinated water. The positivecontrol involved treatment with 3 lg mL�1 of Temephos(O,O 0-(thiodi-4,1-phenylene)bis(O,O-dimethyl phosphoro-thioate)) in dechlorinated water. The LC10, LC50 andLC90 were calculated only for the most active extracts using

Table 1Plants and plant tissues selected on the basis of ethnobotanical information for the screening programme for activity against larvae of Aedes aegypti

Family/species voucherreference

City-state ofcollection anddate of harvest

Plant part assayed/yield (%)and conditions of extraction

Ethnobotanical use, activityand/or medicinal application

Reference

AmaralidaceaeAmaryllis

belladonna LMaceio – AL,08/2003

Bulb 4.4a Diuretic; used to treat bronchitis,asthma and chronic defluxion

Braga (1953)

AnnonaceaeAnnona

crassiflora

Mart

Brasılia – DF,08/1999

Root wood 9.1a, root bark6.0a, stem 18.0a, fruit bark1.3a, pulp 8.1a and seed 15.5a

Molluscicide Martins (1989) andDos Santos andSant’Ana (2001)

JEP 3369 (UB)A. glabra L. Paripueira – AL,

09/2002Seed 15.2a Anthelmintic; anti-rheumatic Correa and Penna (1984)

JEP 3649 (UB)A. montana

Macfad &R. E. Fries

MarechalDeodoro – AL,09/2002

Root 9.7a, wood 12.0a,leaf 7.8a

Anti-tumour, anti-syphilitic;used to treat colic and fever

Correa and Penna (1984)

A. muricata L. Maceio – AL,08/2000

Root 12.0a, stem 9.2a,leaf 10.2a

Anti-parasitic, anti-rheumatic,astringent; emetic

Braga (1953), Gemtchujnicov(1976) and Correa andPenna (1984)

MAC 8530

A. salzmanni A.DC.

Paripueira – AL,08/2000

Fruit bark 12.0a Used to treat tumours andinflammations


JEP 3601 (UB)A. squamosa L. Maceio – AL,

08/2002Root 7.6a, leaf 9.5a,seed 12.0a

Astringent; used to treat gastricdiseases


ApocynaceaeAspidosperma

sp.Sıtio Imburama/Sao Jose da

Stem 7.3a Anti-rheumatic, anti-tumour;anti-diarrhoeic; used to treat fever


JEP 3544 (UB) Tapera – AL, 07/2002

A. tomentosum

MartPlanaltina – GO,08/1996

Root 7.1a

JEP 3417 (UB)Hancornia

speciosa

Lagoa do Abaete– BA, 05/1987

Latex 6.0a Anti-syphilitic; used to treat lungdisease and as a tonic

Braga (1953) andVan Den Berg (1982)

GomesJEP 2022 (UB)Himatanthus sp. Murici – AL, 09/

2000Leaf 11.0a Used to treat malaria and fever Milliken (1997)

MAC 9312H. phaged-

aenicus (Mart.)Brasılia – DF,08/2000

Wood 9.1a Used to treat fever Correa and Penna (1984)

WoodsonJEP 3598 (UB)H. obovatus

(Mull. Arg.)Brasılia – DF,08/1996

Stem 9.6a, leaf 7.2a Used to treat gastric ulcer, hepaticdisease and anthelmintic


R. E. WoodsonJEP 3518 (UB)Mandevilla

hirsuta (Rich)Matriz doCamaragibe –AL, 10/1999

Leaf 8.7a Used to treat snakebite Dos Santos (1996)

K. SchumJEP 3607 (UB)

AraceaeAnthurium affine

SchotPiacabucu – SL,03/2003

Root 9.5a Used to treat vitiligo disease Marcelino Junior (1999)

MAC 17242

AraliaceaeDidymopanax

morototoni

Murici – AL, 07/2000

Wood bark. 8.5a Anthelmintic; larvicidal Dr. Aldenir Feitosa dosSantos (personalcommunication)(Aubl.) Decne &

PlanchJEP 3634 (UB)

(continued on next page)

M.C. de Omena et al. / Bioresource Technology 98 (2007) 2549–2556 2551

Table 1 (continued)





Reference

AsclepiadaceaeCalotropis

procera

Barra de SaoMiguel – AL,06/1982

Whole plant 10.5a Anti-bacterial, anti-malarial,analgesic; anti-diarrhoeic

Olea Gallegos et al. (2002)

(Aiton)W. T. Aiton

MAC 1877

AsteraceaeAcantho-

spermum

Pontal da Barra– AL, 11/1977

Root 7.3a Anti-diarrhoeic; used totreatcough, bronchitis and liver disease

Dr. Jose Elias de Paula(personal communication)

hispidum DC.MAC 00204Senecio

jurgensenii

Eldorado do Sul– RS, 09/1999

Stem 8.5a, flower 9.4a Used to treat malaria and fever Milliken (1997)

Hemsl.JEP 3564 (UB)Vernonia

condensata

Recife – PE, 07/2000

Leaf 6.2b Anti-diarrhoeic; used to treatgastro-intestinal disorders, headache,and protection against snakebites

Monteiro et al. (2001)

BakerJEP 3607 (UB)

BignoniaceaeTabebuia sp. Maceio – AL,

08/2001Leaf 6.3a Astringent, anti-cancer, anti-

inflammatory, anti-allergic,anti- microbial; used to treat wounds

Matos (1994)

Tabebuia

impetiginosa

Maceio – AL,08/2001

Stem bark 6.5a

(Mart exDC.) Standl

BoraginaceaeAustroplenckia

populnea

Planaltina – GO,02/1987

Stem 6.6a Anti-microbial; anti-tumour Dr. Jose Elias de Paula(personal communication)

(Reissek exMart.)Lundell

JEP 1888 (UB)Auxema

oncocalyx

Taub

Monte Pascoal –BA, 04/1987

Stem 10.3a, stem bark 8.5a Tripanocidal activity; used to treatwounds, varicose ulcer and burns

Matos (1994) and Dos Santoset al. (1996)

JEP 3626 (UB)

ConvolvulaceaeIpomea

pescaprae

(L.)

Maceio – AL,05/2000

Stem 7.3a, leaf 6.4a Diuretic, emmenagogue, depurative;used for dressings

Braga (1953)

RothMAC 11073

CurcubitaceaeLuffa cylindrica

(L.)Itabaiana – SE,08/1996

Fruit 6.7a Purgative; used to treat dropsy andfor personal hygiene

Correa and Penna (1984),Vieira (1992) andMatos (1997)M.Roem.

JEP 3639 (UB)Wilbrandia sp. Maceio – AL,

08/1996Root 4.2a Purgative, febrifuge, depurative,

emmenagogue, anti-rheumaticBraga (1953)

JEP 3570 (UB)

LamiaceaeLeonotis

nepetaefolia

Pindoba – AL,07/1982

Root 6.1a, stem 7.0a, leaf 4.5a,fruit 6.3a

Diuretic, febrifuge, anti-asthma,spasmolitic, anti-rheumatic

Martins (1989)

Schimp. ExBenth.

MAC 00175Mentha

pulegium L.Maceio – AL,08/2003

Leaf 6.0a Used to treat cough and huskiness Matos (1994)


Table 1 (continued)





Reference

LeguminoseaeAbarema

cochliacarpus

Barra de SaoMiguel – AL,09/2000

Root 5.6a Used to treat inflammations A.E.G. Santa’Ana(unpublished results)

(Gomes)R. C. Barneby& J.W. Grimes

JEP 3648 (UB)Copaifera

langsdorfii

Commercial oilfrom Belem –PA 05/2000

Commercial Oil Used to treat wounds andblennorrhoea

Braga (1953)

Desf.Commercial oil

Derris sp. Labrea – AM08/1996

Root 18.2a Piscicidal activity Martins (1989)

Erythrina

mulungu Mart.Maceio – AL,08/1986

Root wood 6.3a, stem 3.5a,stem bark 6.1a, leaf 4.6a

Used to treat bronchitis andinflammation

Balbach (1963)

ex. BenthJEP 3593 (UB)Inga edulis Mart Rio Largo – AL,

09/1979Root 15.0a Anti-diarrhoeic; used to treat

woundsBalbach (1963)

MAC 00993Parkia

platycephala

Santa Quiteria –AL, 08/1992

Stem 4.3a Anti-diarrhoeic; used to treatwounds

Martins (1989)

Benth.JEP 3409 (UB)Pterodon

polygalaeflorus

Brasılia – DF,08/2000

Root bark 6.3a, stem 7.2a,stem bark 5.1a, seed 22.0a,pericarp 7.4a

Anti-rheumatic; used to treatthroat infections

Nunan (1985)

(Benth.) Benth.

JEP 3421, 3684,3410 (UB)

Senna

occidentalis (L.)Maceio – AL,02/1981

Seed 5.6a Used to treat malaria Milliken (1997)

Link.MAC 01183

LithraceaeLafoensia pacari

St HilairePlanaltina – GO,05/1998

Stem bark 17.1a Anti-thermic, anti-tumour;used to treat gastric ulcer

Correa and Penna (1984),Lima and Martins (1996)and Sartori and Martins(1996)

JEP 3535 (UB)

MoraceaeChorophora

tinctoria (L.)Maceio – AL,07/1996.

Stem bark 7.6b Used to treat tooth painand wounds


Gaudich. Ex.Benth

Dorstenia sp. Maceio – AL,08/2003

Root 7.2a, stem 6.2a,leaf 6.1a, fruit 5.3a andseed 7.1a

Febrifuge, diuretic Braga (1953)JEP 3603 (UB)

MusaceaeMusa

paradisiaca L.Murici – AL, 08/2003

3.4.a Anti-hemorrhagic; used totreat lung disease

Balbach (1963) andDelorme and Miola (1979)

NictaginaceaeBoerhaavia

coccinea Mill.Maceio –AL, 08/1996

Root 14.6a Used to treat venereal diseaseand urinary system obstruction

Gemtchujnicov (1976),Martins (1989) and Santoset al. (1998)MAC 08512

RhamnaceaeZizyphus

joazeiro Mart.Propia – SE, 09/2000

Stem bark 16.1a Used for mouth and hair hygiene Braga (1953)

JEP 3637 (UB)

(continued on next page)


Table 1 (continued)





Reference

SapindaceaeCupania

oblongifolia

Mart.

Matriz doCamaragibe –AL, 11/1999

Stem 5.1a, stem bark 4.9a,leaf 6.1a

Used to treat cough and whoopingcough


JEP 3538 (UB)Paullinia

meliaefolia JussIbipora – PR,07/1999

Stem 5.1a, stem bark 2.2a,leaf 7.1a

Aphrodisiac; used for rejuvenation De Oliveira et al. (2002)

JEP 3558 (UB)Serjania lethalis A.

St. HilaireMarechalDeodoro – AL,11/1999

Leaf 6.3a Piscicidal activity Correa and Penna (1984)

JEP 3698 (UB)

SapotaceaeManilkara

salzmannii

MarechalDeodoro – AL,11/1994

Stem 7.1a, latex 8.1a Used to treat fever and malaria Milliken (1997)

(A. DC) H. J. LamJEP 3609 (UB)

SimaroubaceaeSimarouba

amara AublItabaiana – SE,09/2001

Stem 7.8a Anti-diarrhoeic; used to treat colic Balbach (1963)

JEP 3640 (UB)

SterculiaceaeBasiloxylon

brasiliensis

K. Schum

Lorena – SP, 11/1994

Wood 6.5a Used for insulation De Paula and Alves (1997)

JEP 3627 (UB)Guazuma

ulmifolia LamNossa Senhorado Socorro – SE,09/2001

Stem bark 6.2a Astringent, depurative,anti-syphilitic; used to treat skindisease

Balbach (1963)

JEP 3644 (UB)

VochysiaceaeVochysia

divergens Pohl.Rio Paraguai,Corumba MS

Leaf 9.1a Used to treat asthma and throatinfections

Hess (1995)

COR/CEUC/UFMS 0500

a Room temperature extraction with ethanol (26–27 �C).b Soxhlet extraction with ethanol.


probit analysis with a reliability interval of 95% (McLaugh-lin et al., 1993).

3. Results and discussion

Fifty-one species, belonging to 42 genera, of Brazilianmedicinal plants were selected, on the basis of publishedethnobotanical information (Table 1), for a screening pro-gramme against fourth instar A. aegypti. The genus Annona

was best represented with six species studied. Of the 84 eth-anolic extracts tested in the screening programme (Table1), only eleven showed LC50 values <200 lg mL�1 againstlarvae of A. aegypti, these being extracts from root bark,root wood and stem of Annona crassiflora, from root, leafand seed of Annona squamosa, from seed of Annona glabra,from root of Annona muricata, from root of a Derris sp.,from stem bark of Erythrina mulungu, and from seed ofPterodon polygalaeflorus. Each of these extracts was sub-

jected to more detailed bioassays in order to determinethe respective LC10, LC50 and LC90 values (Table 2).

The ethanolic extracts of seed of A. glabra and of rootbark of A. crassiflora displayed the highest larvicidal activ-ities with LC50 values of 0.06 and 0.71 lg mL�1, respec-tively. These values are at least one order of magnitudelower than the LC50 (8.54 lg mL�1) of the extract of Derris

root that contains the potent natural insecticides, the rote-nones. Ethanolic extracts of seed of A. squamosa and ofroot wood of A. crassiflora showed activities similar to thatexhibited by Derris root extract, whilst extracts of the rootsof A. squamosa and A. muricata, of the stem of A. crassifl-

ora, and of the seed of P. polygalaeflorus were some 2- to 4-fold less active than the Derris root extract. Least active ofthe extracts studied in detail were those derived from stembark of E. mulungu and from leaves of A. squamosa.

The observed activities against larvae of A. aegypti of theroot bark extract of A. crassiflora (LC50 = 0.71 lg mL�1)and of the seed extract of A. glabra (LC50 = 0.06 lg mL�1)

Table 2Extracts showing significant activities (LC50 <200 lg mL�1) against fourth stage larvae of Aedes aegypti

Family Species Plant tissues screened Larvicidal activitya

LC10 LC50 LC90

Annonaceae Annona crassiflora Root bark 0.09 0.71 5.12Root wood 2.05 8.94 39.0Stem 4.75 16.1 54.8

A. glabra Seed 0.001 0.06 2.75A. muricata Root 8.93 42.3 200A. squamosa Root 15.4 31.9 66.2

Leaf 38.2 169 748Seed 1.22 5.12 21.4

Leguminoseae Derris sp. Root 4.77 8.54 15.2Erythrina mulungu Stem bark 36.9 67.9 125Pterodon polygalaeflorus Seed 20.1 35.7 63.6

a LC values (lg mL�1) are means from four replicates determinations.


are highly comparable with those reported for the pureacetogenins bullatacin (LC50 = 0.10 lg mL�1) and trilobin(LC50 = 0.67 lg mL�1) against Culex larvae (He et al.,1997). These two acetogenins, each bearing adjacent bisTHF rings, have been isolated, respectively, from A. squa-

mosa and Asimia trilobata (He et al., 1997). The activity ofA. squamosa against mosquito larvae (e.g. Culex) (Mehraand Hiradhar, 2000; George and Vincent, 2005) and otherinsects is fairly well-studied and the use of the plant in pestcontrol is protected by patents (Moeschler et al., 1987;Mikolajczak et al., 1988, 1989). Furthermore commercialproducts (Annonin, Annonaine or Anonin) based onextracts of A. squamosa are available on the market for usein the treatment of head lice. However, the activities ofextracts from root bark of A. crassiflora and seed of A.

glabra, screened in the present study, surpass that of A.

squamosa.The results of the screening programme against larvae of

A. aegypti demonstrate that plants of the genus Annona

(family Annonaceae) appear to contain highly active insec-ticidal components. The highest activities were observed inethanolic extracts of seed of A. glabra, followed by extractsof root bark of A. crassiflora and seed of A. squamosa.These extracts will be subjected to activity-guided fraction-ation in our laboratory.

Acknowledgements

This work was financed by the Brazilian funding author-ities, Conselho Nacional de Desenvolvimento Cientıfico eTecnologico (CNPq), Fundacao Coordenacao de Aper-feicoamento de Pessoal de Nıvel Superior (CAPES) andFundacao de Amparo a Pesquisa do Estado de Alagoas(FAPEAL). JSL and MCO wish to thank CNPq andCAPES, respectively, for fellowships. The authors are grate-ful to Rosangela P. de Lira Lemos (Instituto do Meio Ambi-ente do Estado de Alagoas IMA-AL, Maceio AL. Brazil) forhelp with collection/authentication of plant material.

References

Ansari, M.A., Razdan, R.K., Tandon, M., Vasudevan, P., 2000. Larvi-cidal and repellent actions of Dalbergia sissoo Roxb. (F. Legumino-seae) oil against mosquitoes. Bioresour. Technol. 73, 207–211.

Arnason, T., Swain, T., Wat, C.K., Graham, E.A., Partington, S., Towers,G.H.N., Lam, J., 1981. Mosquito larvicidal activity of polyacetylenesfrom species in Asteraceae. Biochem. Syst. Ecol. 9, 63–68.

Balbach, A., 1963. As Plantas Curam, 16th ed. A Verdade Presente, SaoPaulo.

Braga, R., 1953. Plantas do Nordeste Especialmente do Ceara. ColecaoMossoroense, fourth ed. Editora Universitaria-UFRN, Natal.

Chantraine, J.M., Laurent, D., Ballivan, C., Saavedra, G., Ibanez, R.V.S.,1998. Insecticidal activity of essential oil on Aedes aegypti larvae.Phytother. Res. 12, 350–354.

Ciccia, G., Coussio, J., Mongelli, E., 2000. Insecticidal activity againstAedes aegypti larvae of some medicinal South American plants. J.Ethnopharmacol. 72, 185–189.

Consoli, R.A.G.B., Oliveira, R.L., 1994. Principais Mosquitos deImportancia Sanitaria no Brasil. Fiocruz, Rio de Janeiro.

Correa, M.P., Penna, L.A., 1984. Dicionario de Plantas Uteis do Brasil eExoticas Cultivadas. Ministerio da Agricultura-IBDF, Rio de Janeiro.

Delorme, R.T., Miola, H., 1979. Pronto Socorro do Sertao – A Cura pelasPlantas. Grofosul, Porto Alegre.

De Oliveira, J.F., Avila, A.S., Braga, A.C.S., De Oliveira, M.B.N.,Boasquevisque, E.M., Jales, R.L., Cardoso, V.N., Bernardo-Filho, M.,2002. Effect of medicinal plants on the labelling of blood elements withtechnetium-99 m and on the morphology of red blood cells: I Studywith Paullinia cupana. Fitoterapia 73, 305–312.

De Paula, J.E., Alves, J.L.H., 1997. Madeiras Naturais Anatomia,Dentrologia, Dendrometria, Producao e Uso. Fundacao MokitiOkada-MOA, Brasılia.

Dos Santos, E.M.B., 1996. Estudos Preliminares de Enzimologia emArtemisia Annua L. e Determinacao da Estrutura de Pregnano deMandevilla illustris Vell. MSc dissertation, Universidade Federal deAlagoas, Maceio.

Dos Santos, A.F., Sant’Ana, A.E.G., 2001. Molluscicidal properties ofsome species of Annona. Phytomedicine 8, 115–120.

Dos Santos, M.R.G., de Azevedo, D.C., Freitas, L.R., Tonholo, J.,Lemos, T.G., Pedrosa, O.D.L., Goulart, M.O.F., 1996. Estudoeletroquımico da OPB-2, benzoquinona fortemente tripanossomicida.Anais X SIBEE, Sao Carlos-SP, 10, pp. 117–119.

Evans, D.A., Raj, R.K., 1988. Extracts of Indian plants as mosquitolarvicides. Ind. J. Med. Res., 38–41.

Fundacao Nacional de Saude, 1996. Manual de Dengue – VigilanciaEpidemiologica e Atencao ao Doente, second ed. FUNASA –Ministerio da Saude, Brasılia.


Fundacao Nacional de Saude, 2002. Guia de Vigilancia Epidemiologica,vol. 1, fifth ed. FUNASA – Ministerio da Saude, Brasılia.

Gemtchujnicov, I.D., 1976. Manual de Taxonomia Vegetal – Plantas deInteresse Economico, Agrıcola, Ornamentale Medicinal. AgronomicaCeres, Sao Paulo.

George, S., Vincent, S., 2005. Comparative efficacy of Annona squamosa

Linn and Pongiana glabra Vent to Azadirachta indica A. Juss againstmosquitoes. J. Vect. Borne Dis., 159–163.

Green, M.M., Singer, J.M., Sutherland, D.J., Hibben, C.R., 1991.Larvicidal activity of Tagetes minuta. J. Am. Mosquito Control.Assoc. 7, 282–286.

He, K., Zeng, L., Ye, Q., Shi, G., Oberlies, N.H., Zhao, G.X., Njoku, C.J.,McLaughlin, J., 1997. SAR evaluations of Annonaceous acetogeninsfor pesticidal activity. Pesticide Sci. 49, 372–379.

Heal, E.R., Rogers, F.E., 1950. A survey of plants of insecticidal activity.Lloydia 13, 86–162.

Hess, S.C., 1995. Estudos Quımicos, Biologicos e Farmacologicos comVochysia divergens Pohl (Vochysiaceae) e com Ocotea suaveolens

(Meissn.) hassler (Lauraceae). PhD thesis. Universidade Federal deSanta Catarina, Florianopolis.

Lima, G.S., Martins, O.T., 1996. Screening farmacologico de plantasmedicinais utilizadas popularmente como anti-inflamatoria. In: ProcXIV Simp. Plantas Med. do Brasil F-035.

Loset, J.R., Marston, A., Gupta, M.P., Hostettmann, K., 2000. Anti-fungal and larvicidal cordiaquinones from the roots of Cordia

curassavica. Phytochemistry 53, 613–617.Macedo, M.E., Consoli, R.A.G.B., Grandi, T.S.M., Anjos, A.M.G.,

Oliveira, A.B., Mendes, N.M., Queiroz, R.O., Zani, C.L., 1997.Screening of Asteraceae (Compositae) plant extracts for larvicidalactivity against Aedes fluviatilis (Diptera: Culicidae). Mem. Inst.Oswaldo Cruz 92, 565–570.

Marcelino Junior, C.A.C., 1999. Estudo das Atividades Biologicas dePlantas Medicinais. MSc dissertation, Universidade Federal de Ala-goas, Maceio.

Martins, J.E.C., 1989. Plantas Medicinais de Uso na Amazonia, seconded. Graficentro-CEJUP, Belem.

Matos, F.J.A., 1994. Farmacias Vivas, Sistema de Utilizacao de PlantasMedicinais, Projetado para Pequenas Comunidades, second ed.Editora Universitaria-UFC, Fortaleza.

Matos, F.J.A., 1997. O Formulario Fitoterapico do Professor Dias daRocha, second ed. Editora Universitaria-UFC, Fortaleza.

McLaughlin, J., Chang, C.J., Smith, D., 1993. Simple bench-top bioassay(brine shrimp and potato discs) for the discovery of plant antitumorcompounds: reviews of recent progress. In: Kinghorn, A., Balandrin,M. (Eds.), Human Medicinal Agents from Plants, ACS Series No. 534.American Chemical Society, New York.

Mehra, B.K., Hiradhar, P.K., 2000. Effect of crude acetone extract ofseeds of Annona squamosa Linn (Family: Annonaceae) on possible

control potential against larvae of Culex quinquefasciatus Say. J.Entomol. Res., 141–146.

Mikolajczak, K.J., McLaughlin J.L., Rupprecht, J.K., 1988. US Pat., US4721727.

Mikolajczak, K.J., McLaughlin, J.L., Rupprecht, J.K., 1989. US Pat., US4855319.

Milliken, W., 1997. Plants for Malaria – Plants for Fever – MedicinalSpecies in Latin America. A Bibliographic Survey. The Royal BotanicGardens, Kew.

Moeschler, H.F., Pfluger, W., Wendisch, D., 1987. US Pat, US 4.689.232.Monath, T.P., 1994. Yellow fever and dengue – the interactions of virus,

vector and host in the re-emergence of epidemic disease. SeminarsVirol. 5, 133–135.

Monteiro, M.H.D., Gomes, C.M.R., Felzsnswalb, I., Chahoud, I.,Paumgartten, F.J.R., 2001. Toxicological evaluation of a tea fromleaves of Vernonia condensata. J. Ethopharmacol. 74, 149–157.

Neves, D.P., 2000. Parasitologia Humana. Editora Atheneu, Sao Paulo.Nunan, E.A., 1985. Estudo da Atividade Anti-inflamatoria de Furano-

diterpeno Isolados do Pterodon poligaliflorus Benth e de Alguns de seusDerivados. MSc Dissertation. Universidade Federal de Minas Gerais,Belo Horizonte.

Olea Gallegos, R.S., Oliveira, A.V., Silveira, L.M.S., Silveira, E.R., 2002.Organic carbonate from Calotropis procera leaves. Fitoterapia 73, 263–265.

Santos, A.S., Caetano, L.C., Sant’Ana, A.E.G., 1998. A 12a-hydroxyrot-enoid from roots of Boerhaavia coccınea. Phytochemistry 49, 255–258.

Sartori, N.T., Martins, D.T.O., 1996. Screening farmacologico de pantasmedicinais utilizadas popularmente como anti-ulcera em Mato Grosso.In: Proc. XIV Simp. Plantas Med. do Brasil F-092.

Teixeira, G.M., Barreto, L.B., Guerra, Z., 1986. O problema do Aedes

aegypti no Brasil. Rev. Soc. Bras. Med. Trop. 19, 1–3.Teixeira, G.M., Alves, J.L.H., Guerra, Z., 1999. Informe Epidemiologico.

Ministerio da Saude, Brasılia.Vahitha, R., Venkatachalan, M.R., Murugan, K., Jebanesan, A., 2002. A

larvicidal efficacy of Pavonia zeylanica L. and Acacia ferruginea D.Cagainst Culex quinquefasciatus Say. Bioresour. Technol. 82, 203–204.

Van Den Berg, M.E., 1982. Plantas Medicinais na Amazonia – Contrib-uicao ao seu Conhecimento Sistematico. CNPq – PTU/MPEG, Belem.

Vieira, L.S., 1992. Fototerapia da Amazonia – Manual de PlantasMedicinais, second ed. Agronomica Ceres, Sao Paulo.

Wang, Y., Toyota, M., Kranse, F., Hambonge, M., Hostettmann, K.,1990. Polyacetylenes from Artemisia borealis and their biologicalactivities. Phytochemistry 29, 3101–3105.

World Health Organisation, 1981. Instruction for Determining theSusceptibility or Resistance of Mosquito Larvae to Insecticides.WHO – VBC 81-807, pp. 1–6.

Documents

Larvicidal activities against Aedes aegypti of some Brazilian medicinal plants