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      Virulência e multiplicação de isolados de Toxoplasma gondii da região central do Rio Grande do Sul Translated title: Virulence and multiplication of Toxoplasma gondii isolates from the central region of Rio Grande do Sul, Brazil

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          Abstract

          RESUMO: O protozoário Toxoplasma gondii possui a capacidade de infectar diversas espécies animais, geralmente causando distúrbios reprodutivos. Quatro diferentes isolados de T. gondii - Pains #1 (P1), Pains #2 (P2), Santa Flora #1 (SF1) e Santa Flora #306 (SF306) - foram avaliados, após prévia genotipagem. A capacidade de multiplicação e virulência destes isolados foi analisada in vitro (células Vero) e in vivo (camundongos), sendo realizadas 3 passagens para cada isolado em cada modo avaliado, sendo sempre inoculada a dose de 1x104 taquizoítos em todas as passagens. Os camundongos eram observados diariamente, quanto à presença de sinais clínicos e ocorrência de mortalidade após inoculação dos taquizoítos. Os isolados SF1 e SF306, foram os que apresentaram maior multiplicação média do número total de taquizoítos em cada uma das 3 diferentes passagens realizadas para cada um dos isolados tanto in vitro quanto in vivo. Os primeiros sinais clínicos observados nos camundongos ocorreram entre os dias 5 a 11, após inoculação, com mortalidade acontecendo entre os dias 6 a 15, após inoculação. Assim, a multiplicação parasitária in vitro é semelhante à multiplicação in vivo destes isolados de T. gondii; diferentes isolados com o mesmo genótipo apresentam comportamento de virulência semelhante, caracterizando o isolado SF1 como mais virulento para camundongos.

          Translated abstract

          ABSTRACT: The protozoan Toxoplasma gondii has the ability to infect several animal species, usually causing reproductive disorders. Four different isolates of T. gondii - Pains #1 (P1), Pains #2 (P2), Santa Flora #1 (SF1) and Santa Flora #306 (SF306) were evaluated with prior genotyping. Their virulence and multiplication capacity were analyzed in vitro (Vero cells) and in vivo (mice). For each isolate, three passages were performed with dose inoculation 1x104 tachyzoites at each passage. The mice were daily observed to verify of clinical signs and occurrence of mortality after tachyzoites inoculation. The SF1 isolate and SF306 isolate, presented the highest multiplication of the total tachyzoites number in each of the 3 different passages performed in vitro and in vivo. The initial clinical signs in mice were observed between 5 and 11 days, and occurring mortality between 6 and 15 days, after inoculation. Thus, the parasitic multiplication of theses isolates is similar in vitro and in vivo; different isolates within the same genotype have a similar virulence and the SF1 isolate is the most virulent for mice.

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          Most cited references26

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          Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts.

          Infections by the protozoan parasite Toxoplasma gondii are widely prevalent world-wide in animals and humans. This paper reviews the life cycle; the structure of tachyzoites, bradyzoites, oocysts, sporocysts, sporozoites and enteroepithelial stages of T. gondii; and the mode of penetration of T. gondii. The review provides a detailed account of the biology of tissue cysts and bradyzoites including in vivo and in vitro development, methods of separation from host tissue, tissue cyst rupture, and relapse. The mechanism of in vivo and in vitro stage conversion from sporozoites to tachyzoites to bradyzoites and from bradyzoites to tachyzoites to bradyzoites is also discussed.
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            Population structure and mouse-virulence of Toxoplasma gondii in Brazil.

            Recent studies found that isolates of Toxoplasma gondii from Brazil were biologically and genetically different from those in North America and Europe. However, to date only a small number of isolates have been analysed from different animal hosts in Brazil. In the present study DNA samples of 46 T. gondii isolates from cats in 11 counties in São Paulo state, Brazil were genetically characterised using 10 PCR restriction fragment length polymorphism markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. An additional marker, CS3, that locates on chromosome VIIa and has previously been shown to be linked to acute virulence of T. gondii was also used to determine its association to virulence in mice. Genotyping of these 46 isolates revealed a high genetic diversity with 20 genotypes but no clonal Type I, II or III lineage was found. Two of the 46 isolates showed mixed infections. Combining genotyping data in this study with recent reported results from chickens, dogs and cats in Brazil (total 125 isolates) identified 48 genotypes and 26 of these genotypes had single isolates. Four of the 48 genotypes with multiple isolates identified from different hosts and locations are considered the common clonal lineages in Brazil. These lineages are designated as Types BrI, BrII, BrIII and BrIV. These results indicate that the T. gondii population in Brazil is highly diverse with a few successful clonal lineages expanded into wide geographical areas. In contrast to North America and Europe, where the Type II clonal lineage is overwhelmingly predominant, no Type II strain was identified from the 125 Brazil isolates. Analysis of mortality rates in infected mice indicates that Type BrI is highly virulent, Type BrIII is non-virulent, whilst Type BrII and BrIV lineages are intermediately virulent. In addition, allele types at the CS3 locus are strongly linked to mouse-virulence of the parasite. Thus, T. gondii has an epidemic population structure in Brazil and the major lineages have different biological traits.
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              Molecular genotyping of Toxoplasma gondii from Central and South America revealed high diversity within and between populations.

              Recent population studies revealed that a few major clonal lineages of Toxoplasma gondii dominate in different geographical regions. The Type II and III lineages are widespread in all continents and dominate in Europe, Africa and North America. In addition, the type 12 lineage is the most common type in wildlife in North America, the Africa 1 and 3 are among the major types in Africa, and ToxoDB PCR-RFLP #9 is the major type in China. Overall the T. gondii strains are more diverse in South America than any other regions. Here, we analyzed 164 T. gondii isolates from three countries in Central America (Guatemala, Nicaragua, Costa Rica), from one country in Caribbean (Grenada) and five countries from South America (Venezuela, Colombia, Peru, Chile, and Argentina). The multilocous polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based genotyping of 11 polymorphic markers (SAG1, SAG2, alt.SAG2, SAG3, BTUB, GRA6, L358, PK1, C22-8, C29-2 and Apico) were applied to 148 free-range chicken (Gallus domesticus) isolates and 16 isolates from domestic cats (Felis catus) in Colombia; 42 genotypes were identified. Linkage disequilibrium analysis indicated more frequent genetic recombination in populations of Nicaragua and Colombia, and to a lesser degree in populations of Costa Rica and Argentina. Bayesian structural analysis identified at least three genetic clusters, and phylogenetic network analysis identified four major groups. The ToxoDB PCR-RFLP #7, Type III and II were major lineages identified from Central and South America, with high frequencies of the closely related ToxoDB PCR-RFLP #7 and Type III lineages. Taken together, this study revealed high diversity within and between T. gondii populations in Central and South America, and the dominance of Type III and its closely related ToxoDB PCR-RFLP #7 lineages. Copyright © 2011 Elsevier B.V. All rights reserved.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                pvb
                Pesquisa Veterinária Brasileira
                Pesq. Vet. Bras.
                Colégio Brasileiro de Patologia Animal - CBPA. Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) (Rio de Janeiro, RJ, Brazil )
                0100-736X
                1678-5150
                June 2018
                : 38
                : 6
                : 1026-1029
                Affiliations
                [2] Santa Maria Rio Grande do Sul orgnameUniversidade Federal de Santa Maria orgdiv1Centro de Ciências Rurais orgdiv2Departamento de Medicina Veterinária Preventiva Brazil
                Article
                S0100-736X2018000601026
                10.1590/1678-5150-pvb-5472
                98f9162b-d348-4520-a173-b32e89a63e1a

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 12 June 2017
                : 26 June 2017
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 33, Pages: 4
                Product

                SciELO Brazil


                Vero cells,Virulence,Toxoplasma gondii,Rio Grande do Sul,toxoplasmosis,tachyzoites,protozoa,genotypes,parasitoses,células Vero,taquizoítos,Virulência,protozoário,genótipos,toxoplasmose

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