Abstract
The objective of this study was to estimate the Trypanosoma evansi infection rate and epizootical status of wild and domestic animals from the Brazilian Pantanal region using a standardized polymerase chain reaction (PCR). We used a simple DNA extraction method based on Chelex resin (BioRad, USA) on blood eluted from filter paper confetti. Primers directed to repetitive nuclear DNA sequences were used in the PCR, and could detect 30 fg of T. evansi DNA. The analytical specificity of the assay was evaluated using T. evansi, T. rangeli, T. cruzi, Leishmania braziliensis, Crithidia fasciculata and Herpetomonas muscarum DNAs as templates and the technique showed the expected 164 bp specific band solely for Trypanozoon trypanosomes. The application of the standardized PCR protocol in 274 field samples from domestic and wild mammals from the Rio Negro (Brazilian Pantanal region), showed a general infection rate of 10.2% while the traditional parasitological technique (direct search of the protozoan by the microematocrit centrifugue technique) was able to determine infection in only 1.1% of the animals. The peccaries and feral pigs were found to be the animals most frequently infected with T. evansi (24.4% and 30.7%, respectively). Both sampling and extraction methods used herein, showed to be simple and efficient to be applied in epidemiological surveys using PCR.
Similar content being viewed by others
References
Agbo EE, Majiwa PA, Claassen HJ, Te Pas MF (2002) Molecular variation of Trypanosoma brucei subspecies as revealed by AFLP fingerprinting. Parasitology 124:349–358
Claes F, Agbo EC, Radwanska M, Te Pas MF, Baltz T, De Waal DT, Goddeeris BM, Claassen E, Buscher P (2003a) How does Trypanosoma equiperdum fit into the Trypanozoon group? A cluster analysis by RAPD and multiplex-endonuclease genotyping approach. Parasitology 126:425–431
Claes F, Verloo D, De Waal DT, Majiwa PA, Baltz T, Goddeeris BM, Buscher P (2003b) The expression of RoTat 1.2 variable surface glycoprotein (VSG) in Trypanosoma evansi and T equiperdum. Vet Parasitol 116:209–216
Clausen PH, Wiemann A, Patzelt R, Kakaire D, Poetzsch C, Peregrine A, Mehlitz D (1998) Use of a PCR assay for the specific and sensitive detection of Trypanosoma spp in naturally infected dairy cattle in peri-urban Kampala, Uganda. Ann N Y Acad Sci 29:21–31
Davila AM, Herrera HM, Schlebinger T, Souza SS, Traub-Cseko YM (2003) Using PCR for unraveling the cryptic epizootiology of livestock trypanosomosis in the Pantanal, Brazil. Vet Parasitol 117:1–13
De Almeida PJ, Ndao M, Van Meirvenne N, Geerts S (1997) Diagnostic evaluation of PCR in goats experimentally infected with Trypanosoma vivax. Acta Trop 66:45–50
Franke CR, Greiner, M, Mehlitz D (1994) Investigations on naturally occurring Trypanosoma evansi infections in horses, cattle, dogs and capybaras (Hydrochaeris hydrochaeris) in Pantanal de Poconé (Mato Grosso, Brasil). Acta Trop 58:159–169
Gill BS, Singh, J, Gill JS, Kwatra MS (1987) Trypanosoma evansi infection in pigs in India. Vet Rec 120:92
Herrera H M, Dávila AMR, Norek A, Abreu UGP, Souza SS, D’andrea OS, Jansen AM (2004) Enzootiology of Trypanosoma evansi in Pantanal, Brazil. Vet Parasitol 125:263–275
Hoare CA (1972) The Trypanosomes of mammals A zoological monograph. Blackwell Scientific Publications, Oxford
Holland WG, Do TT, Huong NT, Dung NT, Thanh NG, Vercruysse J, Goddeeris BM (2003) The effect of Trypanosoma evansi infection on pig performance and vaccination against classical swine fever. Vet Parasitol 111:115–123
Ijaz MK, Nur-E-Kamal MAS, Mohamed AIA, Dar FK (1998) Comparative studies on the sensitivity of polymerase chain reaction and microscopic examination for the detection of Trypanosoma evansi in experimentally infected mice. Comp Immunol Microbiol Infect Dis 21:215–223
Kanmogne GD, Asonganyi T, Gibson WC (1996) Detection of Trypanosoma brucei gambiense, in serologically positive but aparasitaemic sleeping-sickness suspects in Cameroon, by PCR. Ann Trop Med Parasitol 90:475–483
Katakura K, Lubinga C, Chitambo H, Tada Y (1997) Detection of Trypanosoma congolense and T. brucei subspecies in cattle in Zambia by polymerase chain reaction from blood collected on a filter paper. Parasitol Res 83:241–245
Lourival RFF, da Fonseca GAB (1998) Analise de sustentabilidade do modelo de Caça Tradicional, no Pantanal da Nhecolândia, Corumbá, MS. In: Valladares-Padua C, Bodmer RE, Cullen L Jr (eds) Manejo e Conservação de Vida Silvestre no Brasil. MCT-CNPq, Sociedade Civil Mamirauá
Masiga DK, Smyth AJ, Hayes P, Bromidge TJ, Gibson WC (1992) Sensitive detection of trypanosomes in tsetse flies by DNA amplification. Int J Parasitol 22:909–918
Moser DR, Cook GA, Ochs DE, Baily CP, Mckane MR, Donelson JE (1989) Detection of Trypanosoma congolense and Trypanosoma brucei subspecies by DNA amplification using the polymerase chain reaction. Parasitology 99:57–66
Nantulya VM (1990) Trypanosomiasis in domestic animals: the problem of diagnosis. Rev Sci Tech 9:357–367
Nunes VLB, Oshiro ET (1990). Trypanosoma (Trypanozoon) evansi in the coati from the Pantanal region of Mato Grosso do Sul State, Brazil. Trans R Soc Trop Med Hyg 84:692
Penchenier L, Dumas V, Grebaut P, Reifenberg JM, Cuny G (1996) Improvement of blood and gut processing for PCR diagnosis of trypanosomosis. Parasite 3:387–389
Raina AK, Kumar R, Rajora VS, Singh RP (1985) Oral transmission of Trypanosoma evansi infection in dogs and mice. Vet Parasitol 18:67–69
Reid AS, Husein A, Copeman DB (2001) Evaluation and improvement of parasitological tests for Trypanosoma evansi infection. Vet Parasitol 102:291–297
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Silva RAMS, Barros ATM, Herrera HM (1995) Trypanosomosis outbreaks due to Trypanosoma evansi in the Pantanal, Brazil A preliminary approach on risk factors. Rev Elev Med Vet Pays Trop 48:315–319
Silva RAMS, Victório AM, Ramirez L, Dávila AMR, Trajano V, Jansen AM (1999) Hematological and blood chemistry alterations in coatis (Nasua nasua) naturally infected by Trypanosoma evansi in the Pantanal, Brazil. Rev Elev Med Vet Pays Trop 52:119–122
Solano P, Michel JF, Lefrançois T, De La Rocque S, Sidibé I, Zoungrana A, Cuisance D (1999) Polymerase chain reaction as a diagnosis tool for detecting trypanosomes in naturally cattle in Burkina Faso. Vet Parasitol 86:95–103
Solano P, Jamonneau V, N’guessan P, N’dri L, Dje NN, Miezan TW, Lejon V, Buscher P, Garcia A (2002) Comparison of different DNA preparation protocols for PCR diagnosis of Human African Trypanosomosis in Cote d’Ivoire. Acta Trop 82:349–356
Walsh PS, Metzger PK, Higuchi R (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10:506–513
Wiesenhutter E (1975) Trypanosoma evansi Infections of dogs in Malaysia and considerations of their epidemiology. Southeast Asian J Trop Med Public Health 6:445
Woo PTK (1970) The haematocrit centrifuge technique for the diagnosis of African trypanosomiasis. Acta Trop 27:384–386
Acknowledgements
This work was supported by Conselho Nacional de Pesquisa e Desenvolvimento (CNPq no.470100/01-0); Instituto Oswaldo Cruz/Fiocruz; Programa de Apoio à Pesquisa Estratégica em Saúde/Fiocruz (PAPS III - 0250250108); Conservação Internacional do Brasil and International Atomic Energy Agency (IAEA no.11409/RB). The assistance provided by the local people is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Herrera, H.M., Norek, A., Freitas, T.P.T. et al. Domestic and wild mammals infection by Trypanosoma evansi in a pristine area of the Brazilian Pantanal region. Parasitol Res 96, 121–126 (2005). https://doi.org/10.1007/s00436-005-1334-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-005-1334-6