Universidade de São Paulo

Biblioteca Digital da Produção Intelectual - BDPI

Departamento de Química Fundamental - IQ/QFL Artigos e Materiais de Revistas Científicas - IQ/QFL

2011

Molluscicidal and ovicidal activities of plant

extracts of the Piperaceae on Biomphalaria
glabrata (Say, 1818)

JOURNAL OF HELMINTHOLOGY, v.85, n.1, p.66-72, 2011

http://producao.usp.br/handle/BDPI/16766

Downloaded from: Biblioteca Digital da Produção Intelectual - BDPI, Universidade de São Paulo
Journal of Helminthology (2011) 85, 66–72 doi:10.1017/S0022149X10000258
q Cambridge University Press 2010

Molluscicidal and ovicidal activities of

plant extracts of the Piperaceae on
Biomphalaria glabrata (Say, 1818)
L.N. Rapado1,2*, E. Nakano2, F.P. Ohlweiler4, M.J. Kato3,
L.F. Yamaguchi3, C.A.B. Pereira5 and T. Kawano2
1
Instituto de Ciências Biomédicas, Universidade de São Paulo, Brazil:
2
Lab. Parasitologia e Malacologia, Instituto Butantan, São Paulo, Brazil:
3
Lab. Quı́mica de Produtos Naturais, Universidade de São Paulo, Brazil:
4
Superintendência de Controle de Endemias (SUCEN), São Paulo, Brazil:
5
Depto de Estatı́stica, Núcleo de Bioinformática, Instituto de Matemática,
Universidade de São Paulo, São Paulo, SP, Brazil
(Accepted 16 March 2010; First Published Online 6 May 2010)

Abstract

Schistosomiasis is a tropical disease caused by Schistosoma and occurs in 54

countries, mainly in South America, the Caribbean region, Africa and the eastern
Mediterranean. Currently, 5 to 6 million Brazilian people are infected and 30,000
are under infection risk. Typical of poor regions, this disease is associated with
the lack of basic sanitation and very frequently to the use of contaminated water
in agriculture, housework and leisure. One of the most efficient methods of
controlling the disease is application of molluscicides to eliminate or to reduce
the population of the intermediate host snail Biomphalaria glabrata. Studies on
molluscicidal activity of plant extracts have been stimulated by issues such as
environmental preservation, high cost and recurrent resistance of snails to
synthetic molluscicides. The aim of this study was to determine the molluscicide
action of extracts from Piperaceae species on adult and embryonic stages of
B. glabrata. Fifteen extracts from 13 Piperaceae species were obtained from stems,
leaves and roots. Toxicity of extracts was evaluated against snails at two different
concentrations (500 and 100 ppm) and those causing 100% mortality at 100 ppm
concentration were selected to obtain the LC90 (lethal concentration of 90%
mortality). Piper aduncum, P. crassinervium, P. cuyabanum, P. diospyrifolium and
P. hostmannianum gave 100% mortality of adult snails at concentrations ranging
from 10 to 60 ppm. These extracts were also assayed on embryonic stages of
B. glabrata and those from P. cuyabanum and P. hostmannianum showed 100%
ovicidal action at 20 ppm.

Introduction that 200 million people are infected and 600 million people
are at risk of infection (WHO, 1993; Chitsulo et al., 2000).
Schistosomiasis remains one of the most prevalent Three main schistosome species infect humans:
parasitic infections in the world and has significant Schistosoma mansoni, present mainly in Africa and South
economic and public health consequences. It is estimated America; S. haematobium, found in Africa and the Middle
East; and S. japonicum, which is endemic in Asia.
Freshwater snails of the genus Biomphalaria play a major
*E-mail: [email protected] role as intermediate hosts in the transmission of
 Piperaceae effect on Biomphalaria glabrata 67

S. mansoni. An intense multiplication of parasites occurs from P. aduncum (Orjala et al., 1993a, b) and extracts from
in these snails, making strategies to control snail P. cubeba (Pandey & Singh, 2009).
populations a high priority for reduction of schistoso- In spite of the variety of secondary compounds
miasis transmission in endemic regions (WHO, 1984; described, no systematic evaluation of molluscicidal
Rey, 2008). activity of Piperaceae extracts has been carried out.
At present, niclosamide (Bayluscidew, Bayer, Leverku- Thus, in this work, several crude extracts were assessed
sen, Germany) is the only commercially available regarding molluscicidal and ovicidal properties.
molluscicide recommended by the World Health
Organization (WHO) for large-scale use in schistosomia-
sis control programmes (WHO, 1993). Niclosamide is not Materials and methods
toxic to humans, domestic animals or crops, although it is Molluscicidal and ovicidal assays were performed
costly and ictiotoxic (WHO, 1973; McCullough & Mott, according to the methodology recommended by World
1983). Moreover, it is rapidly decomposed by sunlight Health Organization (WHO, 1965, 1983) and experimen-
and its application does not prevent recolonization of tal procedures were employed according to accepted
sites by surviving snails, which could lead to selection of principles of animal welfare in experimental science.
molluscicide-resistant populations. Thus, novel natural Adults and egg masses were obtained from the
molluscicidal compounds from plants should be inves- population of Biomphalaria glabrata (Say, 1818) originally
tigated as possible alternatives to synthetic products from Belo Horizonte, MG, Brazil and reared under
(Luna et al., 2005). laboratory conditions for several years, fed with fresh
Members of the Piperaceae family are among a wide lettuce ad libitum and a balanced ration.
variety of species under study. The family comprises 14
genera and 1950 species (Mabberley, 1997); Piper and
Peperomia are the most abundant genera, with approxi- Plant material
mately 700 and 600 species, respectively ( Joly, 1993).
Plants from the genus Piper are widely distributed Fresh material of each species (table 1) was collected by
throughout tropical and subtropical regions. Brazil has Dr Massuo Jorge Kato and identified by Dr Elsie
several species and promising studies on biological F. Guimarães from Instituto de Pesquisas Jardim Botânico
properties of plants of the Piperaceae family have been do Rio de Janeiro and vouchers were deposited in the
published (Navickiene et al., 2000; Danelutte et al., 2003, Herbarium of Jardim Botânico do Rio de Janeiro.
2005; Cysne et al., 2005; Yamaguchi et al., 2006).
Phytochemical investigations on Piper species have
Preparation of plant extracts
revealed many bioactive compounds such as amides,
alkaloids, lignans, benzoic acids and chromenes (Parmar Selected parts (table 1) of freshly collected plant
et al., 1997; Alécio et al., 1998; Navickiene et al., 2000; Silva material were separated, immediately air dried and
et al., 2002). Chromenes and benzoic acid derivatives are finally dried in an oven at 458C. Material was ground and
frequently isolated from Piper species showing antimi- powdered materials were extracted with dichlorometha-
crobial, antitumoral, germination inhibition, fungicidal ne:methanol (2:1) at room temperature (25– 278C) three
(Nair & Burke, 1990), insecticidal (Miranda et al., 2002) times and filtered. Extracts were evaporated to dryness
and anti-leishmania activities (Torres-Santos et al., 1999). under vacuum in a rotoevaporator and stored at 2 208C.
In the case of molluscicidal activity, only a few reports A stock solution containing 1000 ppm of each extract
have addressed Piper species, including essential oil from was prepared by suspending 10 mg of extract in 0.1 ml
P. marginatum (Rouquayrol et al., 1980) and P. tuberculatum of 99.9% dimethylsulphoxide (DMSO; Aldrich,
(Sousa & Rouquayrol, 1974), chromenes and benzoic acid Milwaukee, Wisconsin, USA) and making up to 10 ml

Table 1. Piperaceae species screened for molluscicidal activity in B. glabrata.

Species Collecting sites Popular name Sampling date

Piper aduncum L. São Paulo, SP Aperta-ruão, Pimenta-de-macaco 10.1.1999

Piper amplum Kunth São Sebastião, SP Jaborandi 5.12.2000
Piper cuyabanum C.DC. Brotas, SP Pimenta do mato 17.9.1999
Piper crassinervium Kunth Apiaı́, SP Jaborandi 12.2.1999
Piper hoffmanseggianum Rocker & Sehulter Ilha Grande, RJ João-Borandinho 31.12.2000
Piper macedoi Yunck Araraquara, SP 12.2.2001
Peperomia macrostachya (Vahl). A. Dietr. Manaus, AM 28.3.2001
Piper hostmannianum R.S. Manaus, AM 21.3.2001
Piper callosum Ruiz et Pav. Presidente Figueiredo, AM João Brandin 29.3.2001
Piper tuberculatum Jacq. São Paulo, SP Pimenta longa, Pimenta d’arda 19.6.2001
Piper diospyrifolium Kunth São Paulo, SP 1.10.2004
Piper solmsianum C.DC. São Paulo, SP 2.11.2004
Piper corcovadensis Miq.* São Paulo, SP João Brandinho, Falso Jaborandi 3.5.2005

Selected part for all species, leaves; * stem, leaves and roots.
S, São Paulo; RJ, Rio de Janeiro, AM, Amazonas.
68 L.N. Rapado et al.

with dechlorinated water. Stock solutions were diluted containing 1% DMSO under the same experimental
with dechlorinated water in order to provide assay conditions.
solutions.
Data analysis
Assays for molluscicidal/ovicidal activities of plant extracts LC90 was obtained from the logistic regression
Molluscicidal activities of extracts against adults of adjustment using a log-dose transformation. This corre-
B. glabrata were determined according to previously sponds to a maximum likelihood estimated for the
described procedures (WHO, 1965, 1983). Snails with logodds in a multinomial probability model.
10 – 18 mm shell diameter were exposed to test com-
pounds for 24 h. After exposure, snails were washed and
Results
observed daily for 10 days to record the death rate.
A negative control group was maintained in dechlori- All extracts were 100% lethal to B. glabrata at 500 ppm
nated tap water containing 1% DMSO under the same but at 100 ppm, only leaf extracts of P. aduncum,
experimental conditions. Bayluscide WP70w was used as P. crassinervium, P. cuyabanum, P. diospyrifolium and
a positive control. Extracts were tested first at concen- P. hostmannianum showed 100% lethality during 24 h
trations of 500 and 100 ppm and those inactive were not of exposure, so the LC90 (table 2, figs 1 – 3) and
investigated further. LC90 was determined for all selected ovicidal activity (table 3 and fig. 4) of these extracts
extracts with ten animals per dose group, and exper- were determined. Leaf extracts of Pep. macrostachya,
iments were repeated three times. P. hoffmanseggianum, P. macedoi and P. solmsianum did not
Based on values of LC90 for adults, egg masses with show molluscicidal activity at 100 ppm; but mortality
embryos at blastula stage (Camey & Verdonk, 1970) were obtained with extracts of P. corcovadensis (stem, leaf and
exposed to lethal concentrations of extracts for 24 h. At the root), P. amplum (leaf), P. callosum (leaf) and P. tuberculatum
end of exposure, egg masses were washed and observed (stem) reached a maximum of 90% at 100 ppm (table 2).
for mortality daily for 7 days. Assays were repeated three The percentage of dead snails in the control group over
times with about 100 embryos for each concentration. 10 days of observation was 3.3% while dead embryos in
A control group was maintained in dechlorinated water control groups over 7 days were 1.6%.

Table 2. Molluscicidal activity of Piperaceae extracts in adult B. glabrata.

Selected Concentration (%) Mortality LC90 (ppm) [ ] confidence

Species part (ppm) in 10 days intervals

P. aduncum Stem 2.5 37 6.46 [5.34; 9.51]

5 73
10 100
P. crassinervium Stem 20 40 38.15 [33.98; 46.04]
30 73
40 93
50 97
60 100
P. cuyabanum Stem 5 7 12.92 [11.25; 16.98]
10 60
20 100
P. diospyrifolium Stem 5 23 23.30 [19.80; 26.81]
10 27
20 77
30 100
P. hostmannianum Stem 10 20 34.29 [32.13; 36.44]
20 43
30 73
40 100
P. corcovadensisa Stem 100 40 nd
Leaf 100 20 nd
Root 100 90 nd
Pep. macrostachyaa Leaf 100 0 nd
P. ampluma Leaf 100 50 nd
P. callosuma Leaf 100 80 nd
P. hoffmanseggianuma Leaf 100 0 nd
P. macedoia Leaf 100 0 nd
P. solmsianuma Leaf 100 0 nd
P. tuberculatuma Stem 100 20 nd

n ¼ 30 adult snails; Pep., Peperomia.

a
n ¼ 10 adult snails.
 Piperaceae effect on Biomphalaria glabrata 69

(a) 30 (a) 30
Number of dead snails

Number of dead snails

24 24

18 18

12
12
6
6
0
0 2 4 6 8 10 0
Concentration (ppm) 0 5 10 15 20
Concentration (ppm)
(b) 30 (b) 30
Number of dead snails

Number of dead snails

24
24
18
18

12 12

6 6

0 0
0 10 20 30 40 50 60 0 6 12 18 24 30
Concentration (ppm) Concentration (ppm)

Fig. 1. Estimated mortality of B. glabrata exposed to (a) P. aduncum Fig. 2. Estimated mortality of B. glabrata exposed to (a) P.
leaf extract; (b) P. crassinervium leaf extract; logistic tendency cuyabanum leaf extract; (b) P. diospyrifolium leaf extract; logistic
function. tendency function.

Discussion Bayluscide WP70w which induces death only after

24 h, even at 100% lethal concentrations. The efficient
All 15 extracts from Piperaceae species were active with molluscicide action observed in our study requires
100% mortality at 500 ppm. WHO (1983) recommends further investigation in order to unravel the possible
that a crude preparation of plant material should be active mechanism of action for toxicity against B. glabrata.
at 100 ppm or less and kill 90% of snails exposed for 24 h. Among the five selected Piperaceae extracts, P.
Leaf extracts of P. aduncum, P. crassinervium, P. cuyabanum, crassinervium, P. cuyabanum and P. hostmannianum
P. diospyrifolium and P. hostmannianum were active at showed similar activity in adults and embryos.
concentrations of less than 100 ppm, which make Sensitivity was inversely proportional to developmental
them good candidates for natural molluscicides (WHO, stage. Mortality was highest in embryos at blastula stage,
1965, 1983). followed by gastrula, trocophore and veliger stages.
Adult snails showed a similar response to extracts with This higher sensitivity could be attributed to intense
molluscicidal action. A common feature was low values
of LC90 associated with death during 24 h of exposure. At
the highest concentrations, death occurred in the first 30
hours of exposure. Release of haemolymph was also
Number of dead snails

frequently observed and occurred in a dose –response 24

manner. It is quite often associated with rupture of
external membranes, but haemolymph could also be 18
liberated through the haemal pore (Duncan, 1985). At
present, information on the mode of action of mollusci- 12
cides is scarce. As is true for most insecticides, the action
of a molluscicide may conceivably be a multi-component 6
process affecting more than one system. Several
such responses indicative of this have been reported in 0
the literature, for example a reduction in heart rate, 0 10 20 30 40
swelling of tissues and alteration of water balance have Concentration (ppm)
been described (WHO, 1983). In our work, active
extracts caused death after around 4 h of exposure, Fig. 3. Estimated mortality of B. glabrata exposed to P.
differing significantly from the action of the molluscicide hostmannianum leaf extract; logistic tendency function.
70 L.N. Rapado et al.

Table 3. Effect of leaf extracts from P. aduncum, P. cuyabanum, P. crassinervium, P. diospyrifolium and P. hostmannianum on
B. glabrata embryos.

Plant species Stage Concentration (ppm) Number of embryos Dead embryos (%)

P. aduncum Blastula 10 111 54 (48.6)

20 141 69 (48.9)
30 110 71 (64.5)
Gastrula 10 123 31 (25.2)
Trocophore 10 101 6 (5.9)
Veliger 10 110 0
P. cuyabanum Blastula 5 134 0
10 123 66 (53.6)
20 116 116 (100)
Gastrula 10 114 0
20 105 105 (100)
Trocophore 10 127 0
20 120 120 (100)
Veliger 10 105 0
20 106 106 (100)
P. crassinervium Blastula 30 95 4 (4.04)
40 99 16 (16.6)
50 134 134 (100)
Gastrula 50 120 0
Trocophore 50 102 0
Veliger 50 98 0
P. diospyrifolium Blastula 20 98 0
30 142 1 (0.7)
40 187 0
Gastrula 20 113 2 (1.7)
Trocophore 20 99 0
Veliger 20 129 0
P. hostmannianum Blastula 10 120 0
20 133 133 (100)
30 112 112 (100)
40 133 133 (100)
Gastrula 30 90 10 (11.1)
40 90 90 (100)
Trocophore 30 129 2 (1.5)
40 104 104 (100)
Veliger 30 97 1 (0.9)
40 91 91 (100)

cell proliferation activity at early stages; previous In a comprehensive screening, the highest proportions
studies also showed a differential sensitivity accor- of molluscicidal species were detected in Euphorbiaceae,
ding to developmental stages (Kawano et al., 1979; Asteraceae, Leguminosae, Phytolaccaceae, Polygonaceae,
Kawano & Simões, 1987; Kawano et al., 1991; Yamamoto Rubiaceae and Rutaceae species (WHO, 1983; Kloos
et al., 1996). & McCullough, 1987). Jurberg et al. (1989) describe
Piper aduncum was more active in adults than active extracts from two species of Euphorbiaceae. In our
in embryos. A similar profile was observed with study, five out of 13 Piperaceae species fulfilled the
crude extracts and steroidal glycoalkaloid fraction of practical criterion for activity against B. glabrata. Based
Lycopersicon esculentum, which showed high activity in on results obtained with Piper extracts, further studies
adults and low effect in embryos (Leyton et al., 2005). with P. aduncum, P. crassinervium, P. cuyabanum,
Similar results were obtained in B. glabrata, B. alexandrina P. diospyrifolium and P. hostmannianum should be carried
and Bulinus truncatus in a study of the molluscicidal out, looking at the isolation of bioactive compounds.
effect of Phytolacca dodecandra (Lemma et al., 1972). At present, molluscicidal activity of P. aduncum and
Pereira et al. (1978) evaluated a leaf extract of Euphorbia P. diospyrifolium fractions are being assessed. Concomi-
cotinifolia and observed an effect in B. glabrata embryos tantly, we are perfoming bioassays with Daphnia similis
with 48 ppm, a concentration about 20 times higher and Ceridaphnia dubia to evaluate acute and chronic
than the 2.4 ppm observed to have an effect on adult toxicity of bioactive compounds isolated from the
specimens. selected Piper extracts. Cladoceran species have been
Piper diospyrifolium did not show molluscicidal activity used extensively; they have several advantages, such as
in embryos at concentrations evaluated in adults. On the high sensitivity and short reproductive cycles (Farre &
other hand, P. amplum was highly effective in embryos Barcelo, 2003). Inclusion of safety testing will ascertain
(LC90 of 8.8 ppm) despite its low activiy in adults whether toxic effects on non-target organisms are at
(Teixeira, 2004). acceptable levels.
 Piperaceae effect on Biomphalaria glabrata 71

Joly, A.B. (1993) Botânica: introdução à taxonomia vegetal.

11th edn. 777 pp. São Paulo, Nacional.
Jurberg, P., Vasconcellos, M.C. & Mendes, N.M. (1989)
Plantas empregadas como moluscicidas: Uma visão
crı́tica. Memórias do Instituto Oswaldo Cruz 84 (Suppl. I),
76 – 83.
Kawano, T. & Simões, L.C.G. (1987) Morphogenetic
effects of caffeine on Biomphalaria glabrata (Pulmonata,
Planorbidae). Proceedings of the Koninklijke Nederlandse
Akademie van Wetenschappen 90, 281–302.
Kawano, T., Chaves, W.R. & Simões, L.C.G. (1979)
Effects of caffeine on embryonic development
of Biomphalaria glabrata. Ciência e Cultura 31,
1265– 1273.
Kawano, T., Simões, L.C. & Marques, J. (1991)
Biomphalaria glabrata (Say, 1818): Efeitos morfogenéti-
cos do sulfato de nicotina. Revista Brasileira de Biologia
53, 539– 547.
Kloos, H. & McCullough, F.S. (1987) Plants with
recognized molluscicidal activity. pp. 45 – 108 in
Mott, K.E. (Ed.) Plant molluscicides. Chichester, UK,
Fig. 4. Embryos of B. glabrata after exposure at blastula stage to John Wiley & Sons.
10 ppm of P. cuyabanum leaf extract. 1, Dead embryo; 2, normal Lemma, A., Goll, P., Newell, G.W., Parkhurst, R.M. &
embryo at veliger stage. Skinner, W.A. (1972) Studies on the molluscicidal
properties of endod (Phytolaca dodecandra): I. Increased
Acknowledgements potency with butanol extraction. The Journal of
Parasitology 58, 104– 107.
This work was supported by Fundação de Amparo à Leyton, V., Henderson, T.O., Mascara, D. & Kawano, T.
Pesquisa do Estado de São Paulo (FAPESP) and Conselho (2005) Atividade moluscicida de principios ativos
Nacional de Desenvolvimento Cientı́fico e Tecnológico de folhas de Lycopersicon esculentum (Solanales,
(CNPq). Solanaceae) em Biomphalaria glabrata (Gastropoda,
Planorbidae). Iheringia Serie Zoologia 95, 213– 216.
Luna, J.S., Santos, A.F., Lima, M.R.F., Omena, M.C.,
References Mendonça, F.A.C., Bieber, L.W. & Sant’Ana, A.E.G.
Alécio, A.C., Bolzani, V.S., Young, M.C.M., Kato, M.J. & (2005) A study of the larvicidal and molluscicidal
Furlan, M. (1998) Antifungal amide from leaves of activities of some medicinal plants from northeast
Piper hispidum. Journal of Natural Products 61, 637– 639. Brazil. Journal of Ethnopharmacology 97, 199– 206.
Camey, T. & Verdonk, N.H. (1970) The early develop- Mabberley, D.J. (1997) The plant book: a portable dictionary
ment of the snail Biomphalaria glabrata (Say, 1818) and of the vascular plants. 2nd edn. 858 pp. Cambridge,
the origin of the head organs. Netherlands Journal of Cambridge University Press.
Zoology 20, 93 – 121. McCullough, F. & Mott, K.E. (1983) The role of
Chitsulo, L., Engels, D., Montresor, A. & Savioli, L. molluscicides in schistosomiasis control. Bulletin of
(2000) The global status of schistosomiasis and its the World Health Organization 83, 1 – 13.
control. Acta Tropica 77, 41 – 51. Miranda, J.E., Oliveira, J.E.M., Rocha, K.C.G., Bortoli,
Cysne, J.B., Canuto, K.M., Pessoa, O.D., Nunes, E.P. & S.A., Navickiene, H.M.D., Kato, M.J. & Furlan, M.
Silveira, E.R. (2005) Leaf essential oils of four Piper (2002) Potencial inseticida do extrato de Piper
species from the state of Ceará – Northeast of Brazil. tuberculatum sobre Alabama agillacea (Huebner,
Journal of Brazilian Chemical Society 16, 1378–1381. 1818) (Lepidoptera: Noctuidae). Revista Brasileira de
Danelutte, A.P., Lago, J.H.G., Young, M.C.M. & Kato, Oleaginosas e Fibrosas 6, 557– 563.
M.J. (2003) Antifungal flavanones and prenylated Nair, M.G. & Burke, B.A. (1990) Antimicrobial
hydroquinones from Piper crassinervium Kunth. Piper metabolite and related compounds. Journal of
Phytochemistry 64, 555– 559. Agriculture and Food Chemistry 38, 1093– 1096.
Danelutte, A.P., Constantin, M.B., Delgado, G.E., Navickiene, H.M.D., Alécio, A.C., Kato, M.J., Bolzani,
Braz-Filho, R. & Kato, M.J. (2005) Divergency of secon- V.S., Young, M.C.M., Cavalheiro, A.J. & Furlan, M.
dary metabolism in cell suspension cultures and diffe- (2000) Antifungal amides from Piper hispidum and
rentiated plants of Piper cernuum and Piper crassinervium. Piper tuberculatum. Phytochemistry 6, 621– 626.
Journal of Brazilian Chemical Society 16, 1425–1430. Orjala, J., Erdelmeier, C.A.J., Wright, A.D. & Sticher, O.
Duncan, J. (1985) The toxicology of plant molluscicides. (1993a) Two chromenes and a prenylated benzoic
Pharmacology & Therapeutics 27, 243– 264. acid derivate from Piper aduncum. Phytochemistry 34,
Farre, M. & Barcelo, D. (2003) Toxicity testing of 813– 818.
wastewater and sewage sludge by biosensors, bio- Orjala, J., Erdelmeier, C.A.J., Wright, A.D., Rali, T.
assays and chemical analysis. Trends in Analytical & Sticher, O. (1993b) Five new prenylated
Chemistry 22, 299– 310. p-hydroxybenzoic acid-derivatives with antimicrobial
72 L.N. Rapado et al.

and molluscicidal activity from Piper aduncum leaves. Teixeira, E. (2004) Estudo do efeito moluscicida de
Planta Medica 59, 546– 551. extratos de Piperaceae em Biomphalaria glabrata
Pandey, J.K. & Singh, D. (2009) Molluscicidal activity (Say, 1818). Dissertação de mestrado. Coordenadoria
of Piper cubeba Linn., Piper longum Linn. and dos Institutos de Pesquisa da Secretaria de Estado da
Tribulus terrestris Linn. and their combinations against Saúde de São Paulo.
snail Indoplanorbis exustus Desh. Indian Journal of Torres-Santos, E.C., Moreira, D.L., Kaplan, M.A.C.,
Experimental Biology 47, 643– 648. Meirelles, M.N. & Rossi-Bergmann, B. (1999)
Parmar, V.S., Jain, S.C., Bisht, K.S., Jain, R., Taneja, P., Selective effect of 20 ,60 -dihydroxy-40 -methoxychalcone
Jha, A., Tyagi, O.D., Prasad, A.K., Wengel, J., isolated from Piper aduncum on Leishmania amazonensis.
Olsen, C.E. & Boll, P.M. (1997) Phytochemistry of Antimicrobial Agents and Chemotherapy 5, 1234– 1241.
the genus Piper. Phytochemistry 46, 597– 673. WHO (1965) Molluscicide screening and evaluation.
Pereira, J.P., Souza, C.P. & Mendes, N.M. (1978) Bulletin of the World Health Organization 33, 567– 581.
Propriedades moluscicidas da Euphorbia cotinifolia L. Geneva, WHO.
Revista Brasileira de Pesquisas Medicas e Biológicas 11, WHO (1973) Schistosomiasis control. pp. 1–47. Geneva, WHO.
345– 351. WHO (1983) Report of the scientific working group on plant
Rey, L. (2008) Parasitologia. 3rd edn. 930 pp. Rio de molluscicides, vol. 12, pp. 1 – 11. Geneva, WHO.
Janeiro, Guanabara Koogan S.A. WHO (1984) Report of informal consultation on research on
Rouquayrol, M.Z., Fonteles, M.C., Alencar, J.E., Matos, the biological control of snail intermediated hosts. pp. 1 – 41.
J.A. & Craveiro, A.A. (1980) Molluscicidal activity of Geneva, WHO.
essential oils from Northeastern Brazilian plants. WHO (1993) The control of Schistosomiasis: second report of
Revista Brasileira de Pesquisas Medicas e Biológicas 13, the WHO Expert Committee. 86 pp. Geneva, WHO.
135– 143. Yamaguchi, L.F., Lago, H.J.G., Tanizaki, T.M., Masuo, P.
Silva, R.V., Navickiene, H.M.D., Kato, M.J., Bolzani, & Kato, M.J. (2006) Antioxidant activity of prenylated
V.S., Meda, C.I., Young, M.C. & Furlan, M. (2002) hydroquinone and benzoic derivates from Piper
Antifungal amides from Piper arboreum and Piper crassinervium Kunt. Phytochemistry 67, 1838– 1843.
tuberculatum. Phytochemistry 59, 521– 527. Yamamoto, M.M., Kawano, T., Young, M.C.M.,
Sousa, M.P. & Rouquayrol, M.Z. (1974) Atividade Haraguchi, M. & Hiroki, K. (1996) Molluscicidal
moluscicida de plantas do Nordeste brasileiro. Revista activity of three Brazilian plant species. Fitoterapia 57,
Brasileira de Pesquisas Medicas e Biológicas 7, 388– 393. 59 – 62.