Made available in DSpace on 2016-04-28T20:16:23Z (GMT). No. of bitstreams: 1 2007-Renata Cunha Madureira.pdf: 2108717 bytes, checksum: 17e511fe376ace549930ffeb6fed483c (MD5) Previous issue date: 2007-09-17 / Funda??o Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / Spirochetes of the genus Borrelia attack animals and humans all over the world, causing different types of diseases. The unique species of Borrelia known and identified in Brazil which infects bovines and equines is B. theileri, transmitted by Rhipicephalus (Boophilus) microplus. Several studies were been made to identify another spirochete that circulates in the country and infects both animals and humans, providing clinical symptoms regarding Lyme disease (LD). The objectives of this work were to determine the frequency of serum-positive animals for Borrelia burgdorferi stricto sensu in equines of the Maraj? Island and the municipality of Castanhal, State of Par?; to caracterize morfometrically and genotypically isolates of B. theileri from equine and to analyze genotipycally spirochetes the tick Rhipicephalus (Boophilus) microplus, and human, by analysis of partial sequence of the gene 16S rRNA, fla and rpoB. Serum from 105 equines from the Maraj? Island and 103 from Castanhal municipality were collected and analyzed by indirect ELISA test. Fifteen equines (7.2% of the total) were serum-positive, being 10 from the Maraj? Island (9.5%) and five from the Castanhal municipality (4.9%). For the morfometric and genotypic characterization was collected serum of equine from the Rio de Janeiro State. Tick isolate was obtained from supernatant of a cell culture of ticks from the Mato Grosso do Sul State. Human isolate was obtained from blood sample of a woman, from the Serop?dica municipality, RJ, with LDsimile. The PCR was carried through using primers for the gene 16S rRNA, fla and rpoB. The products of amplification were cloned and sequenced. The analysis of the sequences of the isolated of equine for the genes 16S rRNA and fla of 99% (609/611) and 98% (222/226) of identity with sequences of B. theileri, respectively, in set with morfometric analysis (17.2 ? 3.6μm of length; 10 ? 2 espiras) revealed to be the microrganism in question, B. theileri. For the gene rpoB it was observed 92%(250/270) of identity with sequences of B. hermisii. The analysis of the sequences of the tick isolated revealed an identification of 100% (177/177) for the gene 16S rRNA with sequences of B. burgdorferi deposited in "GeneBank", 100% of identity (23/23) for the gene fla with B. theileri and 92% of identity (250/270) for the gene rpoB with sequence of B. hermisii. The analysis of the sequences of the human isolated revealed an identification of 99% (886/889) for the gene 16S rRNA with B. burgdorferi sequences. The presence of antibodies against Borrelia burgdorferi in equines from both places is an indicative of the presence of Borrelia sp., and, despite the lower frequency of infected animals, attention is needed for the possible occurrence of human borreliosis in those areas, considering the importance of this emergent zoonosis. This is the first genotypic characterization of a Brazilian isolate of B. theileri, as well as the first report of species of Borrelia different of B. theileri attack R. (Boophilus) microplus and confirmation of the presence of the genus Borrelia causing LD-simile in Brazil. / Espiroquetas do g?nero Borrelia acometem animais e humanos em todo o mundo determinando diferentes tipos de doen?as. A ?nica esp?cie de Borrelia reconhecida e identificada no Brasil que infecta bovinos e eq?inos ? Borrelia theileri, transmitida por Rhipicephalus (Boophilus) microplus. V?rios estudos t?m sido feitos a fim de se identificar outro espiroquet?deo que circula em nosso meio e infecta animais e humanos, levando a quadro cl?nico semelhante ? borreliose de Lyme (BL). O objetivo do presente trabalho foi determinar a freq??ncia de animais soropositivos para Borrelia burgdorferi sensu stricto em eq?inos da Ilha de Maraj? e munic?pio de Castanhal, Estado do Par?, caracterizar morfometricamente B. theileri isolado de eq?ino e genotipicamente isolados de espiroquetas obtidos de eq?ino, carrapato R. (Boophilus) microplus e humano, por an?lise das seq??ncias parciais dos genes 16S rRNA, fla e rpoB. Coletou-se 105 soros de eq?inos provenientes da Ilha de Maraj? e 103 do munic?pio de Castanhal (n=208), os quais foram analisados pelo ELISA indireto. Observou-se 15 animais soropositivos (7,2%), sendo 10 (9,5%) animais da Ilha de Maraj? e cinco (4,9%) de Castanhal. Para caracteriza??o morfom?trica e genot?pica foi coletado sangue de um eq?ino, proveniente do Estado do Rio de Janeiro. O isolado de carrapato foi obtido a partir de sobrenadante de cultura de c?lulas de carrapatos R. (Boophilus) microplus, proveniente do Estado do Mato Grosso do Sul. O isolado de humano foi obtido do sangue de um indiv?duo do sexo feminino, residente no munic?pio de Serop?dica-RJ, com BLsimile. Realizou-se a t?cnica de PCR utilizando iniciadores para o gene 16S rRNA, fla e rpoB. Os produtos da amplifica??o foram clonados e sequenciados. A an?lise das seq??ncias do isolado de eq?ino para os genes 16S rRNA e fla, 99% (609/611) e 98% (222/226) de identidade com seq??ncias depositadas no GeneBank para B. theileri, respectivamente, em conjunto com a an?lise morfom?trica (17,2 ? 3,6 μm de comprimento; 10 ? 2 espiras) revelou ser o microrganismo em quest?o, B. theileri. Para o gene rpoB observou-se 92%(250/270) de identidade com seq??ncias de B. hermisii. O sequenciamento do isolado de carrapato revelou 100% de identidade (177/177) para o gene 16S rRNA com seq??ncias de B. burgdorferi depositadas no GeneBank , 100% de identidade (23/23) para o gene fla com B. theileri e 92% de identidade (250/270) para o gene rpoB com B. hermisii. O sequenciamento do gene 16S rRNA de sangue humano com sintomatologia para borreliose de Lyme-simile, apresentou identidade de 99% (886/889) com B. burgdorferi. A presen?a de anticorpos hom?logos contra B. burgdorferi em eq?inos na Ilha de Maraj? e munic?pio de Castanhal ? indicativo da presen?a de Borrelia sp. e, apesar da baixa freq??ncia de animais soropositivos ? necess?rio aten??o para ocorr?ncia de borreliose humana nas regi?es estudadas, considerando a import?ncia dessa enfermidade como zoonose emergente. Esta ? a primeira descri??o genot?pica de isolado brasileiro de B. theileri, assim como o primeiro relato de esp?cie de Borrelia diferente de B. theileri infectando R. (Boophilus) microplus e confirma??o da presen?a de espiroquet?deo do g?nero Borrelia causando borreliose de Lyme-simile no Brasil.