Skip to main content
Top
Published in: Malaria Journal 1/2015

Open Access 01-12-2015 | Research

Anti-malarial activity and toxicity assessment of Himatanthus articulatus, a plant used to treat malaria in the Brazilian Amazon

Authors: Valdicley V Vale, Thyago C Vilhena, Rafaela C Santos Trindade, Márlia Regina C Ferreira, Sandro Percário, Luciana F Soares, Washington Luiz A Pereira, Geraldo C Brandão, Alaíde B Oliveira, Maria F Dolabela, Flávio De Vasconcelos

Published in: Malaria Journal | Issue 1/2015

Login to get access

Abstract

Background

Plasmodium falciparum has become resistant to some of the available drugs. Several plant species are used for the treatment of malaria, such as Himatanthus articulatus in parts of Brazil. The present paper reports the phyto-chemistry, the anti-plasmodial and anti-malarial activity, as well as the toxicity of H. articulatus.

Methods

Ethanol and dichloromethane extracts were obtained from the powder of stem barks of H. articulatus and later fractionated and analysed. The anti-plasmodial activity was assessed against a chloroquine resistant strain P. falciparum (W2) in vitro, whilst in vivo anti-malarial activity against Plasmodium berghei (ANKA strain) was tested in mice, evaluating the role of oxidative stress (total antioxidant capacity - TEAC; lipid peroxidation – TBARS, and nitrites and nitrates - NN). In addition, cytotoxicity was evaluated using the HepG2 A16 cell-line. The acute oral and sub-chronic toxicity of the ethanol extract were evaluated in both male and female mice.

Results

Plumieride was isolated from the ethyl acetate fraction of ethanol extract, Only the dichloromethane extract was active against clone W2. Nevertheless, both extracts reduced parasitaemia in P. berghei-infected mice. Besides, a significant reduction in pulmonary and cerebral levels of NN (nitrites and nitrates) was found, as well as in pulmonary TBARS, indicating a reduced oxidative damage to these organs. The ethanol extract showed low cytotoxicity to HepG2 A16 cells in the concentrations used. No significant changes were observed in the in vivo toxicity studies.

Conclusions

The ethanol extract of H. articulatus proved to be promising as anti-malarial medicine and showed low toxicity.
Literature
3.
go back to reference Brasil, Secretaria de Vigilância em Saúde − Ministério da Saúde. Situação epidemiológica da malária no Brasil 2000 a 2011. Bol Epidemiol. 2013;44:1–16. Brasil, Secretaria de Vigilância em Saúde − Ministério da Saúde. Situação epidemiológica da malária no Brasil 2000 a 2011. Bol Epidemiol. 2013;44:1–16.
4.
go back to reference Meshnick SR, Dobson MJ. The history of anti-malarial drugs. In: Rosenthal PJ, editor. Anti-malarial Chemotherapy. Totow: Humana Press; 2001. p. 15–25.CrossRef Meshnick SR, Dobson MJ. The history of anti-malarial drugs. In: Rosenthal PJ, editor. Anti-malarial Chemotherapy. Totow: Humana Press; 2001. p. 15–25.CrossRef
6.
go back to reference Wongsrichanalai C, Pickard AL, Wernsdorfer WH, Meshnick SR. Epidemiology of drug-resistant malaria. Lancet Infect Dis. 2002;2:209–18.CrossRefPubMed Wongsrichanalai C, Pickard AL, Wernsdorfer WH, Meshnick SR. Epidemiology of drug-resistant malaria. Lancet Infect Dis. 2002;2:209–18.CrossRefPubMed
7.
go back to reference Oliveira AB, Dolabela MF, Braga FC, Jacome RLRP, Povoa MM. Plant-derived anti-malarial agents: new leads and efficient phythomedicines. Part I Alkaloid An Acad Bras Cienc. 2009;81:715–40.CrossRef Oliveira AB, Dolabela MF, Braga FC, Jacome RLRP, Povoa MM. Plant-derived anti-malarial agents: new leads and efficient phythomedicines. Part I Alkaloid An Acad Bras Cienc. 2009;81:715–40.CrossRef
8.
go back to reference Dolabela MF, Oliveira SG, Nascimento JMS, Peres JMV, Wagner H, Povoa MM, et al. In vitro anti-plasmodial activity of extract and constituents from E. febrifuga, a plant traditionally used to treat malaria Brazilian amazon. Phytomedicine. 2008;15:367–72.CrossRefPubMed Dolabela MF, Oliveira SG, Nascimento JMS, Peres JMV, Wagner H, Povoa MM, et al. In vitro anti-plasmodial activity of extract and constituents from E. febrifuga, a plant traditionally used to treat malaria Brazilian amazon. Phytomedicine. 2008;15:367–72.CrossRefPubMed
9.
go back to reference Di Stasi LC, Hiruma-Lima CA. Gentianales medicinais. In: Di Stasi LC, Hiruma-Lima CA (orgs.), editor. Plantas medicinais na Amazônia e na Mata Atlântica. São Paulo, Brazil: Editora da UNESP; 2002. p. 375–85. Di Stasi LC, Hiruma-Lima CA. Gentianales medicinais. In: Di Stasi LC, Hiruma-Lima CA (orgs.), editor. Plantas medicinais na Amazônia e na Mata Atlântica. São Paulo, Brazil: Editora da UNESP; 2002. p. 375–85.
10.
go back to reference Van Den Berg ME. Plantas medicinais na Amazônia: contribuição ao seu conhecimento sistemático. Belém, Brazil: CNPq/PRU/MPEG; 1982. Van Den Berg ME. Plantas medicinais na Amazônia: contribuição ao seu conhecimento sistemático. Belém, Brazil: CNPq/PRU/MPEG; 1982.
11.
go back to reference Gilbert B. Produtos naturais industrializáveis da Amazônia. Fitos. 2006;2:30–8. Gilbert B. Produtos naturais industrializáveis da Amazônia. Fitos. 2006;2:30–8.
12.
go back to reference Miranda ALP, Silva JRA, Rezende CM, Neves JS, Parrini SC, Pinheiro MLB, et al. Anti-inflammatory and analgesic activities of the látex containing triterpenes from Himatanthus sucuuba. Planta Med. 2000;66:284–6.CrossRefPubMed Miranda ALP, Silva JRA, Rezende CM, Neves JS, Parrini SC, Pinheiro MLB, et al. Anti-inflammatory and analgesic activities of the látex containing triterpenes from Himatanthus sucuuba. Planta Med. 2000;66:284–6.CrossRefPubMed
13.
go back to reference Elizabetsky E, Castilhos C. Plants used as analgesics by Amazonian caboclos as a basis for selecting plants for investigation. Int J Crude Drug Res. 1990;28:309–20. Elizabetsky E, Castilhos C. Plants used as analgesics by Amazonian caboclos as a basis for selecting plants for investigation. Int J Crude Drug Res. 1990;28:309–20.
14.
go back to reference Milliken W. Traditional anti-malarial medicine in Roraima- Brazil. Econ bot. 1997;51:212–37.CrossRef Milliken W. Traditional anti-malarial medicine in Roraima- Brazil. Econ bot. 1997;51:212–37.CrossRef
15.
go back to reference Kvist LP, Christensen SB, Rasmussen HB, Mejia K, Gonzalez A. Identification and evaluation of Peruvian plants used to treat malaria and leishmaniosis. J Ethnopharmacol. 2006;106:390–402.CrossRefPubMed Kvist LP, Christensen SB, Rasmussen HB, Mejia K, Gonzalez A. Identification and evaluation of Peruvian plants used to treat malaria and leishmaniosis. J Ethnopharmacol. 2006;106:390–402.CrossRefPubMed
16.
go back to reference Spina AP, Bittrich V. Kinoshita: Typifications, new synonyms and a new combination in Himatanthus (Apocynaceae). Taxon. 2013;62:1304–7.CrossRef Spina AP, Bittrich V. Kinoshita: Typifications, new synonyms and a new combination in Himatanthus (Apocynaceae). Taxon. 2013;62:1304–7.CrossRef
17.
go back to reference Woodson RE. Studies in the Apocynaceae. VII - An evaluationof the genera Plumeria L. and Himatanthus Willd. Ann Mo Bot Gard. 1938;25:189–24.CrossRef Woodson RE. Studies in the Apocynaceae. VII - An evaluationof the genera Plumeria L. and Himatanthus Willd. Ann Mo Bot Gard. 1938;25:189–24.CrossRef
18.
go back to reference Lorenzi H, Matos FJA. Himatanthus sucuuba (Sprice) Wood. In: Lorenzi H, Matos FJA, editors. Plantas medicinais no Brasil: nativas e exóticas cultivadas. São Paulo, Brazil: Instituto Plantarum; 2002. p. 68–9. Lorenzi H, Matos FJA. Himatanthus sucuuba (Sprice) Wood. In: Lorenzi H, Matos FJA, editors. Plantas medicinais no Brasil: nativas e exóticas cultivadas. São Paulo, Brazil: Instituto Plantarum; 2002. p. 68–9.
19.
go back to reference Silva JRA, Rezende CM, Pinto AC, Pinheiro MLB, Cordeiro MC, Tamborini E, et al. Ésteres Triterpênicos de Himatanthus Sucuuba (Spruce) Woodson. Quim Nova. 1998;21:702–4.CrossRef Silva JRA, Rezende CM, Pinto AC, Pinheiro MLB, Cordeiro MC, Tamborini E, et al. Ésteres Triterpênicos de Himatanthus Sucuuba (Spruce) Woodson. Quim Nova. 1998;21:702–4.CrossRef
20.
go back to reference Silva JRA, Amaral ACF, Silveira CV, Rezende CM, Pinto AC. Quantitative determination by HPLC of iridoids in the bark and latex of Himatanthus sucuuba. Acta Amazon. 2007;37:119–22. Silva JRA, Amaral ACF, Silveira CV, Rezende CM, Pinto AC. Quantitative determination by HPLC of iridoids in the bark and latex of Himatanthus sucuuba. Acta Amazon. 2007;37:119–22.
21.
go back to reference Goulart HB, Kimura EA, Peres VJ, Couto AS, Duarte FAA, Katzin AM. Terpenes arrest parasite development and inhibit biosynthesis of isoprenoids in Plasmodium falciparum. Antimicrob Agents Chemother. 2004;48:2502–9.CrossRef Goulart HB, Kimura EA, Peres VJ, Couto AS, Duarte FAA, Katzin AM. Terpenes arrest parasite development and inhibit biosynthesis of isoprenoids in Plasmodium falciparum. Antimicrob Agents Chemother. 2004;48:2502–9.CrossRef
22.
go back to reference Amoa-Onguené P, Ntie-Rang F, Lifongo LL, Ndom JC, Sippl W, Mbaze LM. The potential of anti-malarial compounds derived from African medicinal plants. Part I: A pharmacological evaluation of alkaloids and terpenoids. Malar J. 2013;12:449.CrossRefPubMedCentralPubMed Amoa-Onguené P, Ntie-Rang F, Lifongo LL, Ndom JC, Sippl W, Mbaze LM. The potential of anti-malarial compounds derived from African medicinal plants. Part I: A pharmacological evaluation of alkaloids and terpenoids. Malar J. 2013;12:449.CrossRefPubMedCentralPubMed
23.
go back to reference Rattmann YD, Terluk MR, Souza WM, Santos CAM, Biavatti MW, Torres LB, et al. Effects of alkaloids of Himatanthus lancifolius (Muell. Arg.) Woodson, Apocynaceae, on smooth muscle responsiveness. J Ethnopharmacol. 2005;100:268–75.CrossRefPubMed Rattmann YD, Terluk MR, Souza WM, Santos CAM, Biavatti MW, Torres LB, et al. Effects of alkaloids of Himatanthus lancifolius (Muell. Arg.) Woodson, Apocynaceae, on smooth muscle responsiveness. J Ethnopharmacol. 2005;100:268–75.CrossRefPubMed
24.
go back to reference Yubin JI, Miao Y, Bing W, Yao Z. The extraction, separation and purification of alkaloids in the natural medicine. J Chem Pharm Res. 2014;6:338–45. Yubin JI, Miao Y, Bing W, Yao Z. The extraction, separation and purification of alkaloids in the natural medicine. J Chem Pharm Res. 2014;6:338–45.
25.
go back to reference Mosman T. Rapid colorimetry assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol Methods. 1983;65:55–63.CrossRef Mosman T. Rapid colorimetry assay for cellular growth and survival: application to proliferation and cytotoxicity assay. J Immunol Methods. 1983;65:55–63.CrossRef
26.
27.
go back to reference Lambros C, Vanderberg JP. Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol. 1979;65:418–20.CrossRefPubMed Lambros C, Vanderberg JP. Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol. 1979;65:418–20.CrossRefPubMed
28.
go back to reference Makler MT, Ries JM, Williams JA, Bancroft JE, Piper RC, Gibbins BL, et al. Parasite lactate dehydrogenase as an assay for Plasmodium falciparum drug sensitivity. Am J Trop Med Hyg. 1993;48:739–41.PubMed Makler MT, Ries JM, Williams JA, Bancroft JE, Piper RC, Gibbins BL, et al. Parasite lactate dehydrogenase as an assay for Plasmodium falciparum drug sensitivity. Am J Trop Med Hyg. 1993;48:739–41.PubMed
29.
go back to reference Nkhoma S, Molyneux M, Ward S. In vitro anti-malarial susceptibility profile and prcrt/pfmdr-1 genotypes of Plasmodium falciparum field isolates from Malasi. Am J Trop Med Hyg. 2007;76:1107–12.PubMed Nkhoma S, Molyneux M, Ward S. In vitro anti-malarial susceptibility profile and prcrt/pfmdr-1 genotypes of Plasmodium falciparum field isolates from Malasi. Am J Trop Med Hyg. 2007;76:1107–12.PubMed
30.
go back to reference Organization for Economic Cooperation and Development. OECD. Guideline for Testing of Chemicals. Guideline 420: Acute oral Toxicity-Acute Class Method. Paris, France: OECD Publishing; 2001. Organization for Economic Cooperation and Development. OECD. Guideline for Testing of Chemicals. Guideline 420: Acute oral Toxicity-Acute Class Method. Paris, France: OECD Publishing; 2001.
31.
go back to reference Organization for Economic Cooperation and Development. OECD. Guideline for Testing of Chemicals. Guideline 407. Repeated dose 28-day oral toxicity study in rodents. Paris, France: OECD Publishing; 1995. Organization for Economic Cooperation and Development. OECD. Guideline for Testing of Chemicals. Guideline 407. Repeated dose 28-day oral toxicity study in rodents. Paris, France: OECD Publishing; 1995.
32.
go back to reference Peters W. Drug resistance in Plasmodium berghei. I Chloroquine resistance Exp Parasitol. 1965;17:80–9.CrossRef Peters W. Drug resistance in Plasmodium berghei. I Chloroquine resistance Exp Parasitol. 1965;17:80–9.CrossRef
33.
go back to reference Miller N, Rice-Evans C, Davies M, Gopinathan V, Milner A. A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates. Clin Sci. 1993;84:407–12.PubMed Miller N, Rice-Evans C, Davies M, Gopinathan V, Milner A. A novel method for measuring antioxidant capacity and its application to monitoring the antioxidant status in premature neonates. Clin Sci. 1993;84:407–12.PubMed
34.
go back to reference Percario S, Vital ACC, Jablonka F. Dosagem de malondialdeído. Newslab. 1994;2:46–50. Percario S, Vital ACC, Jablonka F. Dosagem de malondialdeído. Newslab. 1994;2:46–50.
35.
go back to reference Green JS, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate nitrite and (15N) nitrate in biological fluids. Anal Biochem. 1982;126:131–8.CrossRefPubMed Green JS, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate nitrite and (15N) nitrate in biological fluids. Anal Biochem. 1982;126:131–8.CrossRefPubMed
36.
go back to reference Barreto AS, Amaral ACF, Silva JRA, Schripsema J, Rezende CM, Pinto AC. Acido 15-desmetilisoplumierídeo, um novo iridóide isolado das cascas de Plumeria rubra e do látex de Himatanthus sucuuba. Quim Nova. 2007;30:1133–5.CrossRef Barreto AS, Amaral ACF, Silva JRA, Schripsema J, Rezende CM, Pinto AC. Acido 15-desmetilisoplumierídeo, um novo iridóide isolado das cascas de Plumeria rubra e do látex de Himatanthus sucuuba. Quim Nova. 2007;30:1133–5.CrossRef
37.
go back to reference Plouvier V, Favre-Bonvin J. Les iridoids et seco-iridoids: repartition, structure, propietes, biosyntese. Phytochemistry. 1971;10:1697–722.CrossRef Plouvier V, Favre-Bonvin J. Les iridoids et seco-iridoids: repartition, structure, propietes, biosyntese. Phytochemistry. 1971;10:1697–722.CrossRef
38.
go back to reference Pereira MM, Jácome RLRP, Alcantara AFC, Alves RB, Raslan DS. Alcalóides indólicos isolados de espécies do gênero Aspidosperma (Apocynaceae). Quim Nova. 2007;30:970–83.CrossRef Pereira MM, Jácome RLRP, Alcantara AFC, Alves RB, Raslan DS. Alcalóides indólicos isolados de espécies do gênero Aspidosperma (Apocynaceae). Quim Nova. 2007;30:970–83.CrossRef
39.
go back to reference Rebouças SO, Grivicich J, Santos MS, Rodriguez P, Gomes MD, Oliveira SQ, et al. Antiproliferative effect of a traditional remedy Himatanthus articulatus bark on human cancer cell lines. J Ethnopharmacol. 2011;137:926–9.CrossRef Rebouças SO, Grivicich J, Santos MS, Rodriguez P, Gomes MD, Oliveira SQ, et al. Antiproliferative effect of a traditional remedy Himatanthus articulatus bark on human cancer cell lines. J Ethnopharmacol. 2011;137:926–9.CrossRef
40.
go back to reference Cazarin KCC, Corrêa CL, Zambrone FAD. Redução, refinamento e substituição do uso de animais em estudos toxicológicos: uma abordagem atual. Braz J Pharm Sci. 2004;40:290–9. Cazarin KCC, Corrêa CL, Zambrone FAD. Redução, refinamento e substituição do uso de animais em estudos toxicológicos: uma abordagem atual. Braz J Pharm Sci. 2004;40:290–9.
41.
go back to reference Kuria KAM, Chepkwony H, Govaerts C, Roets E, Busson R, Witte P, et al. The anti-plasmodial activity of isolates from Ajuga remota. J Nat Prod. 2002;65:789–93.CrossRefPubMed Kuria KAM, Chepkwony H, Govaerts C, Roets E, Busson R, Witte P, et al. The anti-plasmodial activity of isolates from Ajuga remota. J Nat Prod. 2002;65:789–93.CrossRefPubMed
42.
go back to reference Carvalho LH, Brandão MGL, Santos-Filho D, Lopes JLC, Krettli AU. Anti-malarial activity of crude extracts from brazilian plants studied in vivo in Plasmodium berghei infected mice and in vitro against Plasmodium falciparum in culture. Braz J Med Biol Res. 1991;24:1113–23.PubMed Carvalho LH, Brandão MGL, Santos-Filho D, Lopes JLC, Krettli AU. Anti-malarial activity of crude extracts from brazilian plants studied in vivo in Plasmodium berghei infected mice and in vitro against Plasmodium falciparum in culture. Braz J Med Biol Res. 1991;24:1113–23.PubMed
43.
go back to reference Mahajan S, Atkins WM. The chemistry and biology of inhibitors and pro-drugs targeted to glutathione S-transferases. Cell Mol Life Sci. 2005;65:1221–33.CrossRef Mahajan S, Atkins WM. The chemistry and biology of inhibitors and pro-drugs targeted to glutathione S-transferases. Cell Mol Life Sci. 2005;65:1221–33.CrossRef
44.
go back to reference Gillrie MR, Krishnegowda G, Lee K, Buret A, Robbins SMS, Looareesuwan S, et al. Src-family kinase-dependent disruption of endothelial barrier function by Plasmodium falciparum merozoite proteins. Blood. 2007;110:3426–35.CrossRefPubMedCentralPubMed Gillrie MR, Krishnegowda G, Lee K, Buret A, Robbins SMS, Looareesuwan S, et al. Src-family kinase-dependent disruption of endothelial barrier function by Plasmodium falciparum merozoite proteins. Blood. 2007;110:3426–35.CrossRefPubMedCentralPubMed
45.
go back to reference Adhikari KNJ, Burns KEA, Friedrich JO, Granton JT, Cook DJ, Meade MO. Effect of nitric oxide on oxygenation and mortality in acute lung injury: systemactic review and meta-analysis. BMJ. 2007;334:1–8.CrossRef Adhikari KNJ, Burns KEA, Friedrich JO, Granton JT, Cook DJ, Meade MO. Effect of nitric oxide on oxygenation and mortality in acute lung injury: systemactic review and meta-analysis. BMJ. 2007;334:1–8.CrossRef
46.
go back to reference Pino P, Taoufiq Z, Nicheu J, Vouldoukis I, Mazier D. Blood-brain barrier breakdown during cerebral malaria: suicide or murder? J Thromb Haemost. 2005;94:336–40. Pino P, Taoufiq Z, Nicheu J, Vouldoukis I, Mazier D. Blood-brain barrier breakdown during cerebral malaria: suicide or murder? J Thromb Haemost. 2005;94:336–40.
47.
go back to reference Dimmeler S, Haendeler J, Nehls M. Zeiher: Suppression of apoptosis by nitric oxide via inhibition of interleukin-1β–converting enzyme (ICE)-like and cysteine protease protein (CPP)-32–like proteases. J Exp Med. 1997;185:601–8.CrossRefPubMedCentralPubMed Dimmeler S, Haendeler J, Nehls M. Zeiher: Suppression of apoptosis by nitric oxide via inhibition of interleukin-1β–converting enzyme (ICE)-like and cysteine protease protein (CPP)-32–like proteases. J Exp Med. 1997;185:601–8.CrossRefPubMedCentralPubMed
48.
go back to reference Jeong EJ, Lee KY, Kim SH, Sung SH, Kim YC. Cognitive-enhancing and antioxidant activities of iridoid glycosides from Scrophularia buergeriana in scopolamine-treated mice. Eur J Pharmacol. 2008;588:78–84.CrossRefPubMed Jeong EJ, Lee KY, Kim SH, Sung SH, Kim YC. Cognitive-enhancing and antioxidant activities of iridoid glycosides from Scrophularia buergeriana in scopolamine-treated mice. Eur J Pharmacol. 2008;588:78–84.CrossRefPubMed
49.
go back to reference Aremu AO, Amoo SO, Ndhlala AR, Finnie JF, Van Staden J. Antioxidant activity, acetylcholinesterase inhibition, iridoid content and mutagenic evaluation of Leucosidea sericea. Food Chem Toxicol. 2011;49:1122–8.CrossRefPubMed Aremu AO, Amoo SO, Ndhlala AR, Finnie JF, Van Staden J. Antioxidant activity, acetylcholinesterase inhibition, iridoid content and mutagenic evaluation of Leucosidea sericea. Food Chem Toxicol. 2011;49:1122–8.CrossRefPubMed
50.
go back to reference Souza EL, Grangeiro ARS, Bastos IVGA, Rodrigues GCR, Silva MJ, Anjos FBR, et al. Antitumor activity of leaves of Himatanthus drasticus (Mart.) Plumel – Apocynaceae (Janaguba) in the treatment of Sarcoma 180 tumor. Braz J Pharm Sci. 2010;46:199–203.CrossRef Souza EL, Grangeiro ARS, Bastos IVGA, Rodrigues GCR, Silva MJ, Anjos FBR, et al. Antitumor activity of leaves of Himatanthus drasticus (Mart.) Plumel – Apocynaceae (Janaguba) in the treatment of Sarcoma 180 tumor. Braz J Pharm Sci. 2010;46:199–203.CrossRef
51.
go back to reference Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine. 4th ed. USA: Oxford University Press; 2007. p. 851. Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine. 4th ed. USA: Oxford University Press; 2007. p. 851.
Metadata
Title
Anti-malarial activity and toxicity assessment of Himatanthus articulatus, a plant used to treat malaria in the Brazilian Amazon
Authors
Valdicley V Vale
Thyago C Vilhena
Rafaela C Santos Trindade
Márlia Regina C Ferreira
Sandro Percário
Luciana F Soares
Washington Luiz A Pereira
Geraldo C Brandão
Alaíde B Oliveira
Maria F Dolabela
Flávio De Vasconcelos
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2015
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/s12936-015-0643-1

Other articles of this Issue 1/2015

Malaria Journal 1/2015 Go to the issue