Abstract
Triatoma bahiensis Sherlock & Serafim, 1967, T. lenti Sherlock & Serafim, 1967, and T. pessoai Sherlock & Serafim, 1967 were described based on material collected in the Brazilian state of Bahia. These species were later included in the T. brasiliensis complex based on their geographic distribution. Triatoma bahiensis and T. pessoai were subsequently synonymized with T. lenti. However, the phylogenetic position of T. lenti within the T. brasiliensis complex has remained doubtful. This study aims to assess the taxonomic status of T. bahiensis and to infer the phylogenetic relationships between T. lenti, T. bahiensis and the other members of the T. brasiliensis species complex. The identities of the species in concern were confirmed by comparisons with high resolution photos of the respective type materials; lectotypes are designated for T. pessoai and T. bahiensis. Morphological, morphometric, molecular, and cytogenetic approaches as well as experimental crosses were used. The low viability of experimental crosses combined with morphological and morphometric data allow the differentiation of T. bahiensis and T. lenti. Pairwise cyt b sequence divergence between T. lenti and T. bahiensis was 2.5%. Cytogenetic and molecular analyses grouped T. lenti and T. bahiensis as members of the T. brasiliensis complex. These results revalidate the specific status of T. bahiensis.
References
Abad-Franch, F., Pavan, M.G., Jaramillo, N., Palomeque, F.S., Dale, C., Chaverra, D. & Monteiro, F.A. (2013) Rhodnius barretti, a new species of Triatominae (Hemiptera: Reduviidae) from western Amazonia. Memórias do Instituto Oswaldo Cruz, 108, 92–99.
http://dx.doi.org/10.1590/0074-0276130434.
Abalos, J.W. (1948) Sobre híbridos naturales y experimentales de Triatoma. Anales del Instituto de Medicina Regional, 2, 209-223.
Alevi, K.C.C., Mendonça, P.P., Pereira, N.P., Guerra, A.L., Facina, C.H., Rosa, J.A. & Azeredo-Oliveira, M.T.V. (2013) Distribution of constitutive heterochromatin in two species of triatomines: Triatoma lenti Sherlock and Serafim (1967) and Triatoma sherlocki Papa, Jurberg, Carcavallo, Cerqueira & Barata (2002). Infection, Genetics and Evolution, 13, 301–303.
http://dx.doi.org/10.1016/j.meegid.2012.11.011
Alevi, K.C.C., Mendonça, P.P., Succi, M., Pereira, N.P., Rosa, J.A. & Azeredo-Oliveira, M.T.V. (2012) Karyotype and spermatogenesis in Triatoma lenti (Hemiptera: Triatominae), a potential Chagas vector. Genetics and Molecular Research, 11, 4278–4284.
http://dx.doi.org/10.4238/2012.December.17.3
Alevi, K.C.C., Rosa, J.A. & Azeredo-Oliveira, M.T.V. (2014) Cytotaxonomy of the Brasiliensis subcomplex and the Triatoma brasiliensis complex (Hemiptera: Reduviidae: Triatominae). Zootaxa, 3838, 583–589.
http://dx.doi.org/10.11646/zootaxa.3838.5.7
Almeida, C.E., Oliveira, H.L., Correia, N., Dornak, L.L., Gumiel, M., Neiva, V.L., Harry, M., Mendonça, V.J., Costa, J. & Galvão, C. (2012) Dispersion capacity of Triatoma sherlocki, Triatoma juazeirensis and laboratory-bred hybrids. Acta Tropica, 122, 71–79.
http://dx.doi.org/10.1016/j.actatropica.2011.12.001
Almeida, C.E., Pacheco, R.S., Noireau, F. & Costa, J. (2002) Triatoma rubrovaria (Blanchard, 1843) (Hemiptera: Reduviidae) I: isoenzymatic and chromatic patterns of five populations from the State of Rio Grande do Sul, Brazil. Memórias do Instituto Oswaldo Cruz, 97 (6), 829–834.
Arnold, M.L. (1997) Natural Hybridization and Evolution. University Press, Oxford.
Bookstein, F.L. (1990) Introduction to methods of landmark data. In: Rohlf, F.J. & Bookstein, F.L. (Eds.), Proceedings of the Michigan morphometrics workshop. Special Publication number 2. The University of Michigan Museum of Zoology. Ann Arbor, pp. 215–225.
Campos, R., Botto-Mahan, C., Coronado, X., Jaramillo, N., Panzera, F. & Solari, A. (2011) Wing shape differentiation of Mepraia species (Hemiptera: Reduviidae). Infection, Genetics and Evolution, 11, 329–333.
http://dx.doi.org/10.1016/j.meegid.2010.11.002
Chagas, C. (1909) Nova Tripanozomíase humana: estudos sobre a morfologia e o ciclo evolutivo do Schizotrypanum cruzi n. gen. n. sp. agente etiológico de nova entidade mórbida do homem. Memórias do Instituto Oswaldo Cruz, 1, 159–218.
Correia, N., Almeida, C.E., Lima-Neiva, V., Gumiel, M., Lima, M.M., Medeiros, L.M.O., Mendonça, V.J., Rosa, J.A. & Costa, J. (2013) Crossing experiments confirm Triatoma sherlocki as a member of the Triatoma brasiliensis species complex. Acta Tropica, 128, 162–167.
http://dx.doi.org/10.1016/j.actatropica.2013.06.019
Costa, J., Almeida, C.E., Dotson, E., Lins, A., Vinhaes, M.C., Silveira, A.C. & Beard, C.B. (2003) The epidemiologic importance of Triatoma brasiliensis as a Chagas disease vector in Brazil: a revision of domiciliary captures during 1993-1999. Memórias do Instituto Oswaldo Cruz, 98, 443–449.
http://dx.doi.org/10.1590/S0074-02762003000400002
Costa, J., Almeida, J.R., Britto, C., Duarte, R., Marchon-Silva, V. & Pacheco, R. (1998) Ecotopes, natural infection and trophic resources of Triatoma brasiliensis (Hemiptera, Reduviidae, Triatominae). Memórias do Instituto Oswaldo Cruz, 93, 7–13.
http://dx.doi.org/10.1590/S0074-02761998000100002
Costa, J., Almeida, C.E., Dujardin, J.P. & Beard, C.B. (2003b) Crossing experiments detect genetic incompatibility among populations of Triatoma brasiliensis Neiva, 1911 (Heteroptera, Reduviidae, Triatominae). Memórias do Instituto Oswaldo Cruz, 98 (5), 637–639
Costa, J., Argolo, A. & Felix, M. (2006) Redescription of Triatoma melanica Neiva & Lent, 1941, new status (Hemiptera: Reduviidae: Triatominae). Zootaxa, 1385, 47–58.
Costa, J., Bargues, M.D., Neiva, V.L., Lawrence, G.G., Gumiel, M., Oliveira, G., Cabello, P., Lima, M.M., Dotson, E., Provance, D.W. Jr., Almeida, C.E., Mateo, L., Mas-Coma, S. & Dujardin, J.P. (2016) Phenotypic variability confirmed by nuclear ribosomal DNA suggests a possible natural hybrid zone of Triatoma brasiliensis species complex. Infection, Genetics and Evolution, 37, 77–87.
http://dx.doi.org/10.1016/j.meegid.2015.10.025.
Costa, J., Barth, O.M., Marchon-Silva, V., Almeida, C.E., Freitas-Sibajev, M.G. & Panzera, F. (1997a) Morphological studies on the Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae, Triatominae): genital structures and eggs of different chromatic forms. Memórias do Instituto Oswaldo Cruz, 92, 493–498.
http://dx.doi.org/10.1590/S0074-02761997000400009
Costa, J., Correia, N.C., Neiva, V.L., Gonçalves, T.C.M. & Felix, M. (2013) Revalidation and redescription of Triatoma brasiliensis macromelasoma Galvão, 1956 and an identification key for the Triatoma brasiliensis complex (Hemiptera: Reduviidae: Triatominae). Memórias do Instituto Oswaldo Cruz, 108, 785–789.
http://dx.doi.org/10.1590/0074-0276108062013016
Costa, J., Dornak, L.L., Almeida, C.E. & Peterson, A.T. (2014) Distributional potential of the Triatoma brasiliensis species complex at present and under scenarios of future climate conditions. Parasites & Vectors, 7, 238.
http://dx.doi.org/10.1186/1756-3305-7-238
Costa, J. & Felix, M. (2007) Triatoma juazeirensis sp. nov. from the state of Bahia, Northeastern Brazil (Hemiptera: Reduviidae: Triatominae). Memórias do Instituto Oswaldo Cruz, 102, 87–90.
http://dx.doi.org/10.1590/S0074-02762007000100015
Costa, J., Freitas-Sibajev, M.G., Marchon-Silva, V., Pires, M.Q. & Pacheco, R. (1997b) Isoenzymes detect variation in populations of Triatoma brasiliensis (Hemiptera, Reduviidae, Triatominae). Memórias do Instituto Oswaldo Cruz, 92, 459–464.
http://dx.doi.org/10.1590/S0074-02761997000400002
Costa, J. & Marchon-Silva, V. (1998) Período de intermuda e resistência ao jejum de diferentes populações de Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae, Triatominae). Entomologia y Vectores, 5, 23–34.
Costa, J., Peterson, A.T. & Beard, C.B. (2002) Ecological niche modeling and differentiation of populations of Triatoma brasiliensis Neiva, 1911, the most important Chagas disease vector in northeastern Brazil (Hemiptera, Reduviidae, Triatominae). The American Journal of Tropical Medicine and Hygiene, 67, 516–520.
Costa, J., Peterson, A.T. & Dujardin, J.P. (2009) Morphological evidence suggests homoploid hybridization as a possible mode of speciation in the Triatominae (Hemiptera, Heteroptera, Reduviidae). Infection, Genetics and Evolution, 9, 263–270.
http://dx.doi.org/10.1016/j.meegid.2008.12.005
Espínola, H.N. (1971) Reproductive isolation between Triatoma brasiliensis Neiva, 1911 and Triatoma petrochii Pinto & Barreto, 1925 (Hemiptera Reduviidae). Revista Brasileira de Biologia, 31, 277–281.
Frías-Lasserre, D. (2010) A new species and karyotype variation in the bordering distribution of Mepraia spinolai (Porter) and Mepraia gajardoi Frías et al. (Hemiptera: Reduviidae: Triatominae) in Chile and its parapatric model of speciation. Neotropical Entomology, 39, 572–583.
http://dx.doi.org/10.1590/S1519-566X2010000400017
García, B.A., Moriyama, E.N. & Powell, J.R. (2001) Mitochondrial DNA Sequences of Triatomines (Hemiptera: Reduviidae): Phylogenetic Relationships. Journal of Medical Entomology, 38, 675–683.
http://dx.doi.org/10.1603/0022-2585-38.5.675
García, M., Menes, M., Dorn, P.L., Monroy, C., Richards, B., Panzera, F. & Bustamante, D.M. (2013) Reproductive isolation revealed in preliminary crossbreeding experiments using field collected Triatoma dimidiata (Hemiptera: Reduviidae) from three ITS-2 defined groups. Acta Tropica, 128, 714–718.
http://dx.doi.org/10.1016/j.actatropica.2013.09.003
Gonçalves, T.C.M., Almeida, M.D., Jurberg, J. & Lent, H. (1993) Lista dos exemplares-tipo de Triatomíneos depositados na Coleção Entomológica do Instituto Oswaldo Cruz, Rio de Janeiro (Hemiptera: Reduviidae). Memórias do Instituto Oswaldo Cruz, 88, 327–333.
http://dx.doi.org/10.1590/S0074-02761993000200025
Gonçalves, T.C.M., Teves-Neves, S.C., Santos-Mallet, J.R., Carbajal-de-la-Fuente, A.L. & Lopes, C.M. (2013) Triatoma jatai sp. nov. in the state of Tocantins, Brazil (Hemiptera: Reduviidae: Triatominae). Memórias do Instituto Oswaldo Cruz, 108, 429–437.
http://dx.doi.org/10.1590/S0074-0276108042013006
Gurgel-Gonçalves, R., Galvão, C., Costa, J. & Peterson, A.T. (2012) Geographic distribution of Chagas disease vectors in Brazil based on ecological niche modeling. Journal of Tropical Medicine, 705326, 1–15.
http://dx.doi.org/10.1155/2012/705326
Hall, T.A. (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95–98.
Heitzmann-Fontenelle, T. (1984) Bionomia comparativa de triatomíneos. VI-Híbridos de Triatoma brasiliensis Neiva, 1911 X Triatoma lenti, Sherlocki & Serafim, 1967 (Hemiptera, Reduviidade). Memórias do Instituto Butantan, 47, 175–181.
Huelsenbeck, J.P. & Ronquist, F. (2001) MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics, 17, 754–755.
http://dx.doi.org/10.1093/bioinformatics/17.8.754
International Commission on Zoological Nomenclature (1999) International Code of Zoological Nomenclature. Fourth edition. International Trust for Zoological Nomenclature, London, i–xxix + 1–306.
Jurberg, J., Cunha, V., Cailleaux, S., Raigorodschi, R., Lima, M.S., Rocha, D.S. & Moreira, F.F.F. (2013) Triatoma pintodiasi sp. nov. do subcomplexo T. rubrovaria (Hemiptera, Reduviidae, Triatominae). Revista Pan-Amazônica de Saúde, 4, 43–56.
http://dx.doi.org/10.5123/s2176-62232013000100006
Justi, S.A., Russo, C.A.M., Mallet, J.R.S., Obara, M.T. & Galvão, C. (2014) Molecular phylogeny of Triatomini (Hemiptera: Reduviidae: Triatominae). Parasites & Vectors, 7, 149.
http://dx.doi.org/10.1186/1756-3305-7-149
Lent, H. & Wygodzinsky, P. (1979) Revision of the Triatominae (Hemiptera. Reduviidae) and their significance as vectors of Chagas Disease. Bulletin of the American Museum of Natural History, 163, 125–529.
Lucena, D.T. (1970) Estudos sobre a doença de Chagas no Brasil. Revista Brasileira de Malariología e Doenças Tropicais, 22, 3–173.
Lyman, D.F., Monteiro, F.A., Escalante, A.A., Cordon-Rosales, C., Wesson, D.M., Dujardin, J.P. & Beardi, C.B. (1999) Mitochondrial DNA sequence variation among triatomine vectors of Chagas disease. The American Journal of Tropical Medicine and Hygiene, 60, 377–386.
Martínez, F.H., Villalobos, G.C., Cevallos, A.M., Torre, P.L., Laclette, J.P., Alejandre-Aguilar, R. & Espinoza, B. (2006) Taxonomic study of the Phyllosoma complex and other triatomine (Insecta: Hemiptera: Reduviidae) species of epidemiological importance in the transmission of Chagas disease: using ITS-2 and mtCytB sequences. Molecular Phylogenetics and Evolution, 41, 279–287.
http://dx.doi.org/10.1016/j.ympev.2006.05.002
Martínez-Ibarra, J.A., Grant-Guillén, Y., Ventura-Rodríguez, L.V., Osorio-Pelayo, P.D., Macías-Amezcua, M.D., Meillón-Isáis, K., Alejandre-Aguilar, R., Rodríguez-Bataz, E. & Nogueda-Torres, B. (2011) Biological and genetic aspects of crosses between species of the genus Meccus (Hemiptera: Reduviidae Triatominae). Memórias do Instituto Oswaldo Cruz, 106, 293–300.
http://dx.doi.org/10.1590/S0074-02762011000300007
Martínez-Ibarra, J.A., Salazar-Schettino, P.M., Nogueda-Torres, B., Vences, M.O., Tapia-González, J.M. & Espinoza-Gutiérrez, B. (2009) Occurrence of hybrids and laboratory evidence of fertility among three species of the Phyllosoma complex (Hemiptera: Reduviidae) in Mexico. Memórias do Instituto Oswaldo Cruz, 104, 1125–1131.
http://dx.doi.org/10.1590/S0074-02762009000800009
Mas-Coma, S. & Bargues, M.D. (2009) Populations, hybrids and the systematic concepts of species and subspecies in Chagas disease triatomine vectors inferred from nuclear ribosomal and mitochondrial DNA. Acta Tropica, 110, 112–136.
http://dx.doi.org/10.1016/j.actatropica.2008.10.013
Mendonça, V.J., Alevi, K.C.C., Medeiros, L.M.O., Nascimento, J.D., Azeredo-Oliveira, M.T.V. & Rosa, J.A. (2014) Cytogenetic and morphologic approaches of hybrids from experimental crosses between Triatoma lenti Sherlock & Serafim, 1967 and T. sherlocki Papa et al., 2002 (Hemiptera: Reduviidae). Infection, Genetics and Evolution, 26, 123–131.
http://dx.doi.org/10.1016/j.meegid.2014.05.015
Mendonça, V.J., Oliveira, J., Rimoldi, A., Ferreira Filho, J.C.R., Araújo, R.F. & Rosa, J.A. (2015) Triatominae Survey (Hemiptera: Reduviidae: Triatominae) in the South-Central Region of the State of Bahia, Brazil between 2008 and 2013. The American Journal of Tropical Medicine and Hygiene, 92, 1076–1080.
http://dx.doi.org/10.4269/ajtmh.14-0556
Mendonça, V.J., Silva, M.T.A., Araújo, R.F., Martins Jr, J., Bacci Jr, M., Almeida, C.E., Costa, J., Graminha, M.A.S., Cicarelli, R.M.B. & Rosa, J.A. (2009) Phylogeny of Triatoma sherlocki (Hemiptera: Reduviidae: Triatominae) inferred from two mitochondrial genes suggests its location within the Triatoma brasiliensis complex. The American Journal of Tropical Medicine and Hygiene, 81, 858–864.
http://dx.doi.org/10.4269/ajtmh.2009.08-0664
Monteiro, F.A., Barrett, T.V., Fitzpatrick, S., Cordon-Rosales, C., Feliciangeli, D. & Beard, C.B. (2003) Molecular phylogeography of the Amazonian Chagas disease vectors Rhodnius prolixus and R. robustus. Molecular Ecology, 12, 997–1006.
http://dx.doi.org/10.1046/j.1365-294X.2003.01802.x
Monteiro, F.A., Donnelly, M.J., Beard, C.B. & Costa, J. (2004) Nested clade and phylogeographic analyses of the Chagas disease vector Triatoma brasiliensis in Northeast Brazil. Molecular Phylogenetics and Evolution, 32, 46–56.
http://dx.doi.org/doi:10.1016/j.ympev.2003.12.011
Monteiro, F.A., Escalante, A.A. & Beard, C.B. (2001) Molecular tools and triatomine systematics: a public health perspective. Trends in Parasitology, 2001, 17, 344–347.
http://dx.doi.org/10.1016/S1471-4922(01)01921-3
Panzera, F., Pérez, R., Nicolini, P., Hornos, S., Costa, J., Borges, E., Diotaiuti, L. & Schofield, C.J. (2000) Chromosome homogeneity in populations of Triatoma brasiliensis Neiva 1911 (Hemiptera – Reduviidae – Triatominae). Cadernos de Saúde Pública, 16, 83–88.
http://dx.doi.org/10.1590/S0102-311X2000000800009
Panzera, F., Pérez, R., Panzera, Y., Alvarez, F., Scvortzoff, E. & Salvatella, R. (1995) Karyotype evolution in holocentric chromosomes of three related species of triatomines (Hemiptera-Reduviidae). Chromosome Research, 3, 143–150.
http://dx.doi.org/10.1007/BF00710707
Pérez, R., Hernández, M., Quintero, O., Scvortzoff, E., Canale, D., Méndez, L., Cohanoff, C., Martino, M. & Panzera, F. (2005) Cytogenetic analysis of experimental hybrids in species of Triatominae (Hemiptera-Reduviidae). Genetica, 125, 261–270.
http://dx.doi.org/10.1007/s10709-005-0369-z
Poinar Jr., G. (2005) Triatoma dominicana sp. n. (Hemiptera: Reduviidae: Triatominae) and Trypanosoma antiquus sp. n. (Stercoralia: Trypanosomatidae), the first fossil evidence of a Triatominae—Trypanosomatid vector association. Vector-Borne and Zoonotic Diseases, 5, 72–81.
http://dx.doi.org/10.1089/vbz.2005.5.72
Poinar Jr., G. (2013) Panstrongylus hispaniolae sp. n. (Hemiptera: Reduviidae: Triatominae), a new fossil triatomine in Dominican amber, with evidence of gut flagellates. Palaeodiversity, 6, 1–8.
Posada, D. (2008) jModelTest: Phylogenetic Model Averaging. Molecular Biology and Evolution, 25, 1253–1256.
http://dx.doi.org/10.1093/molbev/msn083
Rodrigues, J.M.S., Moreira, F.F.F., Cordeiro, I.R.S., Almeida, M.D. & Jurberg, J. (2015) List of the type-specimens of Reduviidae (Hemiptera: Heteroptera) in the Triatomines Collection of the Oswaldo Cruz Institute, Brazil. Zootaxa, 3936, 181–206.
http://dx.doi.org/10.11646/zootaxa.3936.2.2
Rohlf, F.J. (1996) Morphometric spaces, shape components and the effects of linear transformations. In: Marcus, L.F., Corti, M., Loy, A., Naylor, G. & Slice, D.E. (Eds.), Advances in morphometric. NATO ASI Series A: Life Sciences, Vol. 284. New York: Plenum Publishing, pp. 117–129.
Rohlf, F.J. (1999a) tpsDig, version 1.18. Available from: http://www.life.bio.sunysb.edu/morph/ (accessed July 2015)
Rohlf, F.J. (1999b) tpsRelw, version 1.18. Available from: http://www.life.bio.sunysb.edu/morph/ (accessed July 2015)
Rosa, J.A., Rocha, C.S., Gardim, S., Pinto, M.C., Mendonça, V.J., Ferreira Filho, J.C.R., Carvalho, E.O.C., Camargo, L.M.A., Oliveira, J., Nascimento, J.D., Cilense, M. & Almeida, C.E. (2012) Description of Rhodnius montenegrensis n. sp. (Hemiptera: Reduviidae: Triatominae) from the state of Rondônia. Brazil. Zootaxa, 3478, 62–76.
Sambrook, J., Fritsch, E.F. & Maniatis, T. (1989) Molecular Cloning: a Laboratory Manual. Vols. 1–3. Cold Spring Harbor: Cold Spring Harbor Laboratory Press,
Schofield, C.J. & Galvão, C. (2009) Classification, evolution, and species groups within the Triatominae. Acta Tropica, 110, 88–100.
http://dx.doi.org/10.1016/j.actatropica.2009.01.010
Schreiber, G., Pinho, A.C., Parentoni, R., Salgado, A.D. & Godoy Jr, T.L. (1974) Cytogenetics of triatominae: II-Spermatogenesis in hybrid between Triatoma infestans and Triatoma pseudomaculata (Hemiptera, reduviidae). Revista Brasileira de Biologia, 34, 1–9.
Sherlock, I.A. & Serafim, E.M. (1967) Triatoma lenti sp.n., Triatoma pessoai sp.n. e Triatoma bahiensis sp.n. do Estado da Bahia. Brasil (Hemiptera, Reduviidae). Gazeta Médica da Bahia, 67, 75–92.
Sherlock, I.A. & Serafim, E.M. (1972) Fauna Triatominae do Estado da Bahia. Brasil. I – As espécies e distribuição geográfica. Revista da Sociedade Brasileira de Medicina Tropical, 6, 265–276.
http://dx.doi.org/10.1590/S0037-86821972000500005
Thompson, J.D., Higgins, D.G. & Gibson, T.J. (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, 22, 4673–4680.
http://dx.doi.org/10.1093/nar/22.22.4673
Usinger, R.L., Wygodzinsky, P. & Ryckman, E.R. (1966) The biosystematics of Triatominae. Annual Review of Entomology, 11, 309–329.
http://dx.doi.org/10.1146/annurev.en.11.010166.001521
Vinhaes, M.C., Oliveira, S.V., Reis, P.O., Sousa, A.C.L., Silva, R.A., Obara, M.T., Bezerra, C.M., Costa, V.M., Alves, R.V. & Gurgel-Gonçalves, R. (2014) Assessing the vulnerability of Brazilian municipalities to the vectorial transmission of Trypanosoma cruzi using multi-criteria decision analysis. Acta Tropica, 137, 105–110.
http://dx.doi.org/10.1016/j.actatropica.2014.05.007
WHO. (2013) Chagas disease (American trypanosomiasis). Available from: http://www.who.int/mediacentre/factsheets/fs340/en/index.html (accessed January 2014)