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Doctoral Thesis
DOI
10.11606/T.10.2016.tde-14082015-170043
Document
Author
Full name
Samantha Yuri Oshiro Branco Valadas
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Soares, Rodrigo Martins (President)
Brandão, Paulo Eduardo
Gregori, Fabio
Madeira, Alda Maria Backx Noronha
Nishi, Sandra Mayumi
Title in Portuguese
Caracterização molecular de isolados de Sarcocystis spp. obtidos de marsupiais do gênero Didelphis spp. pela análise de gene mitocondrial, gene de apicoplasto, espaçador interno transcrito (ITS-1) e genes codificadores de antígenos de superfície (SAGs)
Keywords in Portuguese
Barcoding DNA
SAG
Sarcocystis falcatula
Sarcocystis neurona
Sarocystidea
Brasil
Abstract in Portuguese
Em um trabalho anterior, foi avaliada a variabilidade de Sarcocystis spp. isolados de gambás oriundos do estado do Rio Grande do Sul pesquisando sequências gênicas codificadoras de antígenos de superfície (SAGs). Os resultados indicaram haver linhagens de isolados de Sarcocystis (relacionadas geneticamente a S. falcatula) que trocam genes em prováveis processos de recombinação sexual. A proposta deste estudo foi de conhecer as variações gênicas e relações filogenéticas em Sarcocystis spp. isolados de gambás através da análise de loci gênicos de genoma mitocondrial (CytB), de genoma de apicoplasto (ClpC) e das regiões espaçadoras transcritas internas (ITS-1), comparando a diversidade encontrada nestes grupos com a observada em integrantes da sub-familia Toxoplasmatinae. E também de conhecer a diversidade de genes codificadores de SAG-2, SAG-3 e SAG-4. Neste estudo foram obtidos esporocistos de Sarcocystis spp. através de raspado intestinal e fezes provenientes de gambás do gênero Didelphis, oriundos do Estado de São Paulo, Rio Grande do Sul e Rio Grande do Norte. Os marcadores moleculares CytB, ClpC e ITS-1 revelaram uma ampla variabilidade genotípica e alelos inéditos entre os isolados do Brasil. Os resultados levam a crer que é possível a existência de espécies “híbridas” de Sarcocystis no Brasil, com mistura de genes de ambas as espécies. Em relação aos genes codificadores de SAGs, foi encontrada uma diversidade ainda maior do que trabalho similar realizado. Os resultados sugerem uma possível ocorrência de reassortment e recombinação de alelos em sequências SAGs, contribuindo ainda mais para a geração de variabilidade e consequentemente na configuração antigênica do parasito. Estes resultados demonstram que isolados brasileiros possuem uma composição genética diferente do reportado em demais localidade, o que pode ter reflexos importantes no conhecimento da diversidade das espécies de Sarcocystis que infectam gambás em nosso meio e em toda a epidemiologia das infecções produzidas pelos protozoários deste grupo
Title in English
Molecular characterization of Sarcocystis spp. isolates from marsupials of the genus Didelphis spp. through the analysis of mitochondrial and apicoplast genes, internal transcribed spacer region (ITS-1) and surface antigen genes (SAGs)
Keywords in English
Sarcocystis falcatula
Sarcocystis neurona
Brazil
SAG. DNA Barcoding
Sarocystidea
Abstract in English
A great variability in surface antigens genes (SAGs) of Sarcocystis spp. sporocysts was found in a previous study, through the intestinal scraping technique of opossums from the south region of Brazil. The results indicated probable sexual recombination between lineages of Sarcocystis isolates (genetically related to Sarcocystis falcatula). The aim of this research is the study of genetic variability and phylogenetic relationships among Sarcocystis spp. isolates from captured opossums in Brazil, by the analysis of molecular markers less subjected to selective pressure, such as DNA barcode, for this purpose, genetic loci from mitochondrial (CytB) and apicoplast genome (ClpC), and internal transcribed spacer regions (ITS-1) were applied and compare the results with the diversity found within members of the Toxoplasmatinae sub-family. The diversity among the SAG-2, SAG-3 and SAG-4 genes was also studied. The Sarcocystis sporocysts were obtained from opposum’s intestinal scraping and faeces from São Paulo State, Rio Grande do Sul State and Rio Grande do Norte State, Brazil. The molecular markers CytB, ClpC and ITS-1 revealed a vast genotypic variability and novel alleles among the Brazilian isolates. The results indicate possible existence of hybrid species of Sarcocystis in Brazil, with a gene mixture from both species. Regarding the SAG genes, an even greater variability was found than previously reported. The results also suggest the occurrence of reassortment and allele recombination of SAG sequences, contributing even more to the variability and thus in the parasites antigenic configuration. In this study, the results demonstrate that isolates from Brazil have a different genetic composition than reported in other locations, which may have important impacts in the knowledge of the diversity of Sarcocystis species shed by opossums in our environment and in all of the epidemiology of infections of protozoa of this particular group
 
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Publishing Date
2016-04-14
 
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