Genotyping Characterization of Toxoplasma gondii in Cattle, Sheep, Goats and Swine from the North of Portugal
AbstractBackground: Epidemiological investigations on Toxoplasma gondii infection have found a significant association between human toxoplasmosis and consumption of raw or undercooked meat. The present study aimed to characterize genotypes of T. gondii in 20 cattle, 40 sheep, 15 goats and 16 pigs from the North of Portugal.Methods: Nested PCR amplified the surface antigen 2 (SAG2) gene. Sequencing analysis was performed in order to assess the prevalence of SAG2 type strains (I, II and III).Results: Three and 4 strains of SAG2 type II were identified in heart samples of cattle and sheep, respectively. Three SAG2 type II strains were detected in brain, diaphragm and heart of 3 pigs. Three strains detected in heart samples of 3 goats belonged to SAG2 types I or II; with the same result being observed in heart samples from 2 sheep and in 2 brain and 1 heart samples from 3 pigs.Conclusion: SAG2 type II has been shown for the first time to infect cattle in North of Portugal. In addition, SAG2 type II has also been confirmed as the predominant strain in sheep and pigs in the same region. This is the first molecular report of T. gondii in goats from Portugal.
Dubey JP. Toxoplasmosis of Animals and Hu-mans. 2nd ed. Boca Raton, FL, USA: CRC Press; 2010.
Chanton-Greutmann H, Thoma R, Corboz L, Borel N. Abortion in small ruminants in Swit-zerland: investigations during two lambing sea-sons (1996-1998) with special regard to chla-ydial abortions. Pospischil A Schweiz Arch Tierheilkd. 2002; 144:483–492.
Dubey JP. Toxoplasmosis in sheep-the last 20 years. Vet Parasitol. 2009; 162: 1–14. doi: 10.1016/j.vetpar.2009.02.026.
Dubey JP. Toxoplasmosis in pigs-the last 20 years. Vet Parasitol. 2009; 164: 89–103. doi: 10.1016/j.vetpar.2009.05.018.
Ajzenberg D, Bañuls AL, Tibayrenc M, Dardé ML. Microsatellite analysis of Toxoplasma gondii population shows considerable polymorphism structured into two main clonal groups. Int J Parasitol. 2000; 32: 27–38.
Dubey JP, Vianna M.B, Sousa S, Canada N, Meireles S, Correia da Costa JM, Marcet PL, Lehmann T, Dardé ML, Thulliez P. Character-ization of Toxoplasma gondii isolates in free-range chickens from Portugal. J Parasitol. 2006; 92: 184–186.
Esteves ABGVB. Caracterização genética de estirpes de Toxoplasma gondii isoladas de produtos humanos e animais. MSc, University of Lisbon, Lisbon, Portugal, 2006.
Sousa S, Ajzenberg D, Canada N, Freire, L., Da Costa JM, Dardé ML, Thulliez P, Dubey JP. Biologic and molecular characterization of Tox-oplasma gondii isolates from pigs from Portugal. Vet Parasitol. 2006; 135: 133–136.
Dumètre A, Ajzenberg D, Rozette L, Mercier A, Dardé ML. Toxoplasma gondii infection in sheep from Haute-Vienne, France: seropreva-lence and isolate genotyping by microsatellite analysis. Vet Parasitol. 2006; 142: 376–379.
Montoya A, Miró G, Mateo M, Ramírez C, Fuentes I. Molecular characterization of Toxo-plasma gondii isolates from cats in Spain. J Para-sitol. 2008; 94: 1044–1046.
Herrmann DC, Pantchev N, Vrhovec MG, Barutzki D, Wilking H, Fröhlich A, Lüder CGK, Conraths FJ, Schares G. Atypical Toxo-plasma gondii genotypes identified in oocysts shed by cats in Germany. Int J Parasitol. 2010; 40: 285–292. doi: 10.1016/j.ijpara.2009.08.001.
Berger-Schoch AE, Herrmann DC, Schares G, Müller N, Bernet D, Gottstein B, Frey CF. Prevalence and genotypes of Toxoplasma gondii in feline faeces (oocysts) and meat from sheep, cattle and pigs in Switzerland. Vet Parasitol. 2011; 177: 290–297. doi:
Lopes AP, Dubey JP, Neto F, Rodrigues A, Martins T, Rodrigues M, Cardoso L. Seroprev-alence of Toxoplasma gondii infection in cattle, sheep, goats and pigs from the North of Por-tugal for human consumption. Vet Parasitol. 2013; 193: 266–269. doi: 10.1016/j.vetpar.2012.12.001.
Miller S, Dykes D, Polesky H. A simple salting out procedure for extracting DNA from hu-man nucleated cells. Nucleic Acids Res. 1988; 16: 1215-1219.
Howe DK, Honoré S, Derouin F, Sibley LD. Determination of genotypes of Toxoplasma gondii strains isolated from patients with toxo-plasmosis. J Clin Microbiol. 1997; 35: 1411–1414.
Cassaing S, Bessières MH, Berry A, Berrebi A, Fabre R, Magnaval JF. Comparison between two amplification sets for molecular diagnosis of toxoplasmosis by real-time PCR. J Clin Mi-crobiol. 2006; 44: 720–724.
Lopes AP, Dubey JP, Moutinho O, Gargaté MJ, Vilares A, Rodrigues M, Cardoso L. Seroepidemiology of Toxoplasma gondii infection in women from the North of Portugal in their childbearing years. Epidemiol Infect.2012; 140: 872–877. doi: 10.1017/S0950268811001658.
Moré G, Basso W, Bacigalupe D, Venturini MC, Venturini L. Diagnosis of Sarcocystis cruzi, Neospora caninum, and Toxoplasma gondii infec-tions in cattle. Parasitol Res. 2008;102: 671–675.
Aspinall TV, Marlee D, Hyde JE, Sims PF. Prevalence of Toxoplasma gondii in commercial meat products as monitored by polymerase chain reaction–food for thought? Int J Parasitol. 2002; 32: 1193–1199.
Dubey JP, Thulliez P. Persistence of tissue cysts in edible tissues of cattle fed Toxoplasma gondii oocysts. Am J Vet Res. 1993; 54: 270–273.
Belfort-Neto R, Nussenblatt V, Rizzo L, Muc-cioli C, Silveira C, Nussenblatt R, Khan A, Si-bley LD, Belfort RJr. High prevalence of unu-sual genotypes of Toxoplasma gondii infection in pork meat samples from Erechim, Southern Brazil. An Acad Bras Cienc. 2007; 79: 111–114.
Fuentes I, Rubio JM, Ramírez C, Alvar J. Gen-otypic characterization of Toxoplasma gondii strains associated with human toxoplasmosis in Spain: direct analysis from clinical samples. J Clin Microbiol. 2001; 39: 1566–1570.
Ferreira IM, Idal JE, Costa-Silva TA, Meira CS, Hiramoto RM, Penalva de Oliveira AC, Pe-reira-Chioccola VL. Toxoplasma gondii: genotyp-ing of strains from Brazilian AIDS patients with cerebral toxoplasmosis by multilocus PCR-RFLP markers. Exp Parasitol. 2008; 118: 221–217.
Su C, Zhang X, Dubey JP. Genotyping of Tox-oplasma gondii by multilocus PCR-RFLP mark-ers: a high resolution and simple method for identification of parasites. Int J Parasitol. 2006; 36: 841–848.
Dubey JP, Thulliez P, Powell EC. Toxoplasma gondii in Iowa sows: comparison of antibody ti-ters to isolation of T. gondii by bioassays in mice and cats. J Parasitol. 1995; 81: 48–53.
Gajadhar AA, Aramini JJ, Tiffin G, Bisaillon J-R. Prevalence of Toxoplasma gondii in Canadian market-age pigs. J Parasitol. 1998; 84: 759–763.
Damriyasa IM, Bauer C, Edelhofer R, Failing K, Lind P, Petersene E, Schares G, Tenter AM, Volmer R, Zahner H. Cross-sectional survey in pig breeding farms in Hesse, Germany: sero-prevalence and risk factors of infections with Toxoplasma gondii, Sarcocystis spp. and Neospora caninum in sows. Vet Parasitol. 2004; 126: 271–286.
Weiss LM, Kim K. The development and biol-ogy of bradyzoites of Toxoplasma gondii. Front Biosci. 2000; 5: D391–D405.
Waldeland H. Toxoplasmosis in sheep. The prevalence of Toxoplasma antibodies in lambs and mature sheep from different parts of Nor-way. Acta Vet Scand. 1976; 17: 432–440.
Mason S, Quinnell RJ, Smith JE. Detection of Toxoplasma gondii in lambs via PCR screening and serological follow-up. Vet Parasitol. 2010; 169: 258–263. doi: 10.1016/j.vetpar.2010.01.021.