Applications of polymerase chain reaction for the detection of equine Leishmania sp. infection
DOI:
https://doi.org/10.5433/1679-0359.2020v41n1p199Keywords:
Leishmania infantum, Diagnostic methods, Infection, Horses.Abstract
Leishmaniasis is a neglected zoonotic disease caused by a variety of pathogenic Leishmania species. In the New World, especially in Brazil, visceral leishmaniasis (VL) is caused by Leishmania infantum. The pathogen can infect several animal species including dogs, foxes, rodents, primates, felines, equines and humans. Dogs act as the primary domestic reservoirs. This study aimed to use polymerase chain reaction (PCR) for detecting Leishmania infection in horses living in a canine visceral leishmaniasis (CVL) endemic region. DNA samples from horse peripheral blood were used to perform PCR. Templates were amplified using primers for the kinetoplast DNA (kDNA) minicircles, which were able to detect different species of Leishmania. In addition, primers for internal transcribed spacer (ITS) of ribosomal DNA were used for detection of Trypanosomatidae sp. Amongst the 75 (39%) positive PCR samples from total 192 samples, 21 samples were positive for kDNA and 63 samples were positive for either ITS, ITS1, or ITS2 gene markers. The kDNA PCR and sequencing allowed the detection of L. infantum in horse blood samples. To our knowledge, this is the first report of equine infection with L. infantum in Southern Brazil. These results proved that L. infantum could also infect horses in addition to humans and dogs, as well as in European countries. This conclusion emphasizes the urgent need to follow up investigation of the infection in these animals.Downloads
References
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Belo, V. S., Werneck, G. L., Barbosa, D. S., Simões, T. C., Nascimento, B. W. L., Silva, E. S. da, & Struchiner, C. J. (2013). Correction: factors associated with visceral leishmaniasis in the americas: a systematic review and meta-analysis. PLoS Neglected Tropical Diseases, 7(5). doi: 10.1371/annotation/83856044-8747-4d93-9a1e-64f16bb60c07
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Bigeli, J. G., Oliveira Júnior, W. P. de, & Teles, N. M. M. (2012). Diagnosis of Leishmania (Leishmania) chagasi infection in dogs and the relationship with environmental and sanitary aspects in the municipality of Palmas, state of Tocantins, Brazil. Revista da Sociedade Brasileira de Medicina Tropical, 45(1),18-23. doi: 10.1590/S0037-86822012000100005
Ministério da Saúde. (2014). Manual de vigilância e controle da leishmaniose visceral. Brasília: Ministério da Saúde - Divisão de Vigilância Epidemiológica.
Brito, C. F. P. C., & Pita-Pereira, D. (2014). Diagnóstico molecular de Leishmania spp. em Flebótomos provenientes de áreas de ocorrência de leishmanioses. In F. da Conceição-Silva & C. R. Alves. Leishmanioses do continente americano (Chap. 13, pp. 217-231). Rio de Janeiro: Editora FIOCRUZ.
Cortes, S.; Rolão, N.; Ramada, J., & Campino, L. (2004). PCR as a rapid and sensitive tool in the diagnosis of human and canine leishmaniasis using Leishmania donovani sl-specific kinetoplastid primers. Transactions of the Royal Society of Tropical Medicine and Hygiene, 98(1), 12-17. doi: 10.1016/S0035-9203(03)00002-6
Dantas-Torres, F. (2009). Canine leishmaniosis in South America. Parasites & Vectors, 2(1), S1. doi: 10.1186/1756-3305-2-S1-S1
Deboni, S. C., Barbosa, M., & Ramos, R. R. (2011). Leishmaniose Visceral no Rio Grande do Sul. Boletim Epidemiológico, 13 (1), 1-8. Retrived from: https://www.cevs.rs.gov.br/upload/arquivos/ 201812/27124724-n-1-marco.pdf
Echchakery, M., Chicharro, C., Boussaa, S., Nieto, J., Carrillo, E., Sheila, O.,… Boumezzough, A. (2017). Molecular detection of Leishmania infantum and Leishmania tropica in rodent species from endemic cutaneous leishmaniasis areas in Morocco. Parasites & Vectors, 10(1), 454-462. doi: 10.1186/s13071-017-2398-8
El Tai, N., Osman, O., El Fari, M., Presber, W., & Schönian, G. (2000). Genetic heterogeneity of ribosomal internal transcribed spacer in clinical samples of Leishmania donovani spotted on filter paper as revealed by single-strand conformation polymorphisms and sequencing. Transactions of the Royal Society of Tropical Medicine and Hygiene, 94(5), 575-579. doi: 10.1016/S0035-9203(00)90093-2
El Tai, N. O., El Fari, M., Mauricio, I., Miles, M. A., Oskam, L., El Safi, S. H.,… Schönian, G. (2001). Leishmania donovani: intraspecific polymorphisms of Sudanese isolates revealed by PCR-based analyses and DNA sequencing. Experimental Parasitology, 97(1), 35-44. doi: 10.1006/expr.2001.4592
Escobar, T. A., Döwich, G., Zuravski, L., Cantele, L. C., Duarte, C. A., & Lübeck, I. (2018). Risk factors associated to canine visceral leishmaniasis in Uruguaiana city, Brazil. Semina: Ciências Agrárias, 39(1), 211-219. Retrived from http://dx.doi.org/10.5433/1679-0359.2018v39n1p211
Feitosa, F. L. F., Leal, J., Mendes, L. C. N., Peiro, J. R., Perri, S. H. V., Lima, V. M. F. de,... Marcondes, M. (2012). Estudo soroepidemiológico de leishmaniose em equinos na região de Araçatuba-SP, Brasil, área endêmica para leishmaniose visceral. Brazilian Journal of Veterinary Research and Animal Science, 49(6), 500-502. Retrived from http://dx.doi.org/10.11606/issn.1678-4456.v49i6p500-502
Fernández-Bellon, H., Solano-Gallego, L., Bardagí, M., Alberola, J., Ramis, A., & Ferrer, L. (2006). Immune response to Leishmania infantum in healthy horses in Spain. Veterinary Parasitology, 135(2),181-185. Retrived from https://dx.doi.org/1016/j.vetpar.2005.09.007
Figueiredo, F., & Madeira, M. de F. (2014). Os parasitos e a questão da infecção em animais domésticos e domiciliados. In F. da Conceição-Silva & C. R. Alves. Leishmanioses do continente americano (Chap. 15, pp. 259-273). Rio de Janeiro: Editora FIOCRUZ.
Gama, A., Elias, J., Ribeiro, A. J., Alegria, N., Schallig, H. D., Silva, F.,... Cotovio, M. (2014). Cutaneous leishmaniosis in a horse from northern Portugal. Veterinary Parasitology, 200(1), 189-192. Retrived from https://dx.doi.org/10.1016/j.vetpar.2013.12.005
Koehler, K., Stechele, M., Hetzel, U., Domingo, M., Schönian, G., Zahner, H., & Burkhardt, E. (2002). Cutaneous leishmaniosis in a horse in southern Germany caused by Leishmania infantum. Veterinary Parasitology, 109(1), 9-17. doi: 10.1016/S0304-4017(02)00246-7
Kuhls, K., Mauricio, I. L., Pratlong, F., Presber, W., & Schönian, G. (2005). Analysis of ribosomal DNA internal transcribed spacer sequences of the Leishmania donovani complex. Microbes and Infection, 7(11-12), 1224-1234. doi: doi.org/10.1016/j.micinf.2005.04.009
Lainson, R., & Rangel, E. F. (2005). Lutzomyia longipalpis and the eco-epidemiology of American visceral leishmaniasis, with particular reference to Brazil: a review. Memórias do Instituto Oswaldo Cruz, 100(8), 811-827. doi: 10.1590/S0074-02762005000800001
le Fichoux, Y., Quaranta, J.-F., Aufeuvre, J.-P., Lelievre, A., Marty, P., Suffia, I.,… Kubar, J. (1999). Occurrence of Leishmania infantum parasitemia in asymptomatic blood donors living in an area of endemicity in southern France. Journal of Clinical Microbiology, 37(6), 1953-1957.
Lombardi, M. C., Turchetti, A. P., Tinoco, H. P., Pessanha, A. T., Soave, S. A., Malta, M. C.,… Santos, R. L. (2014). Diagnosis of Leishmania infantum infection by polymerase chain reaction in wild mammals. Pesquisa Veterinária Brasileira, 34(12), 1243-1246. doi: 10.1590/S0100-736X2014001200017
Lombardo, G., Pennisi, M. G., Lupo, T., Chicharro, C., & Solano-Gallego, L. (2014). Papular dermatitis due to Leishmania infantum infection in seventeen dogs: diagnostic features, extent of the infection and treatment outcome. Parasites & Vectors 7(1), 120-131. doi: 10.1186/1756-3305-7-120
Lopes, E., Sevá, A., Ferreira, F., Nunes, C., Keid, L., Hiramoto, R., . . . Galvis-Ovallos, F. (2017). Serological and molecular diagnostic tests for canine visceral leishmaniasis in Brazilian endemic area: one out of five seronegative dogs are infected. Epidemiology & Infection, 145(12), 2436-2444. Retrived from https://doi-org.ez96.periodicos.capes.gov.br/10.1017/S0950268817001443
Lukeš, J., Guilbride, D. L., Votýpka, J., Zíková, A., Benne, R., & Englund, P. T. (2002). Kinetoplast DNA network: evolution of an improbable structure. Eukaryotic cell, 1(4), 495-502. doi: 10.1128/EC.1.4.495-502.2002
Macedo-Silva, V. P., Martins, D. R., Queiroz, P. V. S. de., Pinheiro, M. P. G., Freire, C. C., Queiroz, J. W., . . . Jeronimo, S. M. (2014). Feeding preferences of Lutzomyia longipalpis (Diptera: Psychodidae), the sand fly vector, for Leishmania infantum (Kinetoplastida: Trypanosomatidae). Journal of Medical Entomology 51(1), 237-244. doi: 10.1603/ME12131
Müller, N., Hentrich, B., Frey, C. F., & Welle, M. (2015). Quantitative PCR for the diagnosis of cutaneous leishmaniasis from formalin-fixed and paraffin-embedded skin sections. Molecular and Cellular Probes, 29(6), 507-510. doi: 10.1016/j.mcp.2015.09.008
Müller, N., Welle, M., Lobsiger, L., Stoffel, M. H., Boghenbor, K. K., Hilbe, M.,… Von Bomhard, W. (2009). Occurrence of Leishmania sp. in cutaneous lesions of horses in Central Europe. Veterinary Parasitology, 166(3), 346-351. doi: 10.1016/j.vetpar.2009.09.001
Pandey, K., Pant, S., Kanbara, H., Shuaibu, M. N., Mallik, A. K., Pandey, B. D.,… Yanagi, T. (2008). Molecular detection of Leishmania parasites from whole bodies of sandflies collected in Nepal. Parasitology Research, 103(2), 293-297. doi: 10.1007/s00436-008-0967-7
Pita-Pereira, D., Lins, R., Oliveira, M. P., Lima, R. B., Pereira, B. A., Moreira, O. C.,... Britto, C. (2012). SYBR Green-based real-time PCR targeting kinetoplast DNA can be used to discriminate between the main etiologic agents of Brazilian cutaneous and visceral leishmaniasis. Parasites & Vectors, 5(1), 5-15. doi: 10.1186/1756-3305-5-15
Sambrook, J., & Russel, D. W. (2001). Molecular cloning: a laboratory manual (Rev. ed.). (Chap. 1, pp. 21-27). New York: Cold Spring Harbor: Cold spring harbor laboratory press.
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2020-01-10
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Escobar, T. A., Döwich, G., Cantele, L. C., Góss, G. C., Porciúncula, M. L., Leite, C. T., … Lubeck, I. (2020). Applications of polymerase chain reaction for the detection of equine Leishmania sp. infection. Semina: Ciências Agrárias, 41(1), 199–212. https://doi.org/10.5433/1679-0359.2020v41n1p199
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