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      Genetic characterization of Toxoplasma gondii from Brazilian wildlife revealed abundant new genotypes

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          Highlights

          • Toxoplasma gondii infections in animals are frequent, and the role of wildlife in its epidemiology is not well understood.

          • Samples from 226 wild animals from different Brazilian localities were submitted to mice bioassay and screened by PCR.

          • A total of 17 genotypes were identified, 13 identified for the first time and four already reported in published literature.

          • Most of genotypes here identified are different from previous studies in domestic animals and humans from Brazil.

          Abstract

          This study aimed to isolate and genotype T. gondii from Brazilian wildlife. For this purpose, 226 samples were submitted to mice bioassay and screened by PCR based on 18S rRNA sequences. A total of 15 T. gondii isolates were obtained, including samples from four armadillos (three Dasypus novemcinctus, one Euphractus sexcinctus), three collared anteaters ( Tamandua tetradactyla), three whited-lipped peccaries ( Tayassu pecari), one spotted paca ( Cuniculus paca), one oncilla ( Leopardus tigrinus), one hoary fox ( Pseudalopex vetulus), one lineated woodpecker ( Dryocopus lineatus) and one maned wolf ( Chrysocyon brachyurus). DNA from the isolates, originated from mice bioassay, and from the tissues of the wild animal, designated as “primary samples”, were genotyped by PCR–restriction fragment length polymorphism (PCR/RFLP), using 12 genetic markers (SAG1, SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L258, PK1, CS3 and Apico). A total of 17 genotypes were identified, with 13 identified for the first time and four already reported in published literature. Results herein obtained corroborate previous studies in Brazil, confirming high diversity and revealing unique genotypes in this region. Given most of genotypes here identified are different from previous studies in domestic animals, future studies on T. gondii from wildlife is of interest to understand population genetics and structure of this parasite.

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          Most cited references35

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          SplitsTree: analyzing and visualizing evolutionary data.

          D Huson (1998)
          Real evolutionary data often contain a number of different and sometimes conflicting phylogenetic signals, and thus do not always clearly support a unique tree. To address this problem, Bandelt and Dress (Adv. Math., 92, 47-05, 1992) developed the method of split decomposition. For ideal data, this method gives rise to a tree, whereas less ideal data are represented by a tree-like network that may indicate evidence for different and conflicting phylogenies. SplitsTree is an interactive program, for analyzing and visualizing evolutionary data, that implements this approach. It also supports a number of distances transformations, the computation of parsimony splits, spectral analysis and bootstrapping.
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            Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii.

            The development of simple, sensitive and rapid methods for the detection and identification of Toxoplasma gondii is important for the diagnosis and epidemiological studies of the zoonotic disease toxoplasmosis. In the past 2 decades, molecular methods based on a variety of genetic markers have been developed, each with its advantages and limitations. The application of these methods has generated invaluable information to enhance our understanding of the epidemiology, population genetics and phylogeny of T. gondii. However, since most studies focused solely on the detection but not genetic characterization of T. gondii, the information obtained was limited. In this review, we discuss some widely used molecular methods and propose an integrated approach for the detection and identification of T. gondii, in order to generate maximum information for epidemiological, population and phylogenetic studies of this key pathogen.
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              Highly Endemic, Waterborne Toxoplasmosis in North Rio de Janeiro State, Brazil

              In Campos dos Goytacazes, northern Rio de Janeiro state, Brazil, reports of uveitis consistent with toxoplasmosis led to a survey of the prevalence and risk factors for Toxoplasma gondii infection in 1997–1999. The survey population was selected randomly from schools, randomly chosen communities, and an army battalion. Serum samples from 1,436 persons were tested. With results adjusted for age, 84% of the population in the lower socioeconomic group was seropositive, compared with 62% and 23% of the middle and upper socioeconomic groups, respectively (p<0.001). When multivariate analysis was performed, drinking unfiltered water was found to increase the risk of seropositivity for the lower socioeconomic (odds ratio [OR]: 3.0, 95% confidence interval [CI] 1.3 to 6.9) and middle socioeconomic (OR: 1.7, 95% CI 1.2 to 2.3) populations. We also found a high T. gondii seroprevalence in this Brazilian community. Drinking unfiltered water increased the risk of T. gondii seropositivity, indicating the potential importance of oocyst transmission in water in this region.
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                Author and article information

                Contributors
                Journal
                Int J Parasitol Parasites Wildl
                Int J Parasitol Parasites Wildl
                International Journal for Parasitology: Parasites and Wildlife
                Elsevier
                2213-2244
                13 October 2014
                13 October 2014
                December 2014
                : 3
                : 3
                : 276-283
                Affiliations
                [a ]Departamento de Medicina Veterinária Preventiva e Saúde Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Avenida Professor Doutor Orlando Marques de Paiva, 87, Cidade Universitária, São Paulo, SP 05508-270, Brazil
                [b ]Curso de Medicina Veterinária das Faculdades Metropolitanas Unidas (UniFMU), Rua Ministro Nelson Hungria, 541, São Paulo, SP 05690-050, Brazil
                [c ]Instituto de Biodiversidade e Floresta, Universidade Federal do Oeste do Pará, Rua Vera Paz, Salé, Santarém, PA 68035-110, Brazil
                [d ]Faculdade de Medicina Veterinária, Universidade Federal de Uberlândia, Avenida Pará, 1720, Campus Umuarama, Uberlândia, MG 38400-902, Brazil
                [e ]Departamento de Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Via de acesso Prof. Paulo Donato Castellane, s/n, Campus de Jaboticabal, Jaboticabal, SP CEP 14049-900, Brazil
                [f ]Instituto Brasileiro para Medicina da Conservação, Tríade, Recife, PE CEP 52061-030, Brazil
                [g ]Parque Estadual Dois Irmãos, Recife, PE CEP 52171-011, Brazil
                [h ]Department of Microbiology, The University of Tennessee, Knoxville, TN 37996-0845, USA
                Author notes
                [* ]Corresponding author. Faculty of Veterinary Medicine, University of São Paulo, Department of Preventive Veterinary Medicine and Animal Health, Av. Prof. Orlando Marques de Paiva, Cidade Universitáira, 05508-270, Sâo Paulo, SP Brazil. Tel.: +55 1130917654; fax: +55 1130917928. sgennari@ 123456usp.br
                Article
                S2213-2244(14)00030-3
                10.1016/j.ijppaw.2014.09.003
                4241539
                25426424
                f2999f26-d473-490a-8365-d0a182136d93
                © 2014 The Authors

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

                History
                : 17 July 2014
                : 19 September 2014
                : 24 September 2014
                Categories
                Article

                t. gondii,isolation,genotyping,pcr/rflp,genetic markers,south america

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