Made available in DSpace on 2016-04-28T20:16:21Z (GMT). No. of bitstreams: 1 2007- Franziska Huber.pdf: 2677706 bytes, checksum: 65e703599b63ae016e9aa85d1752e357 (MD5) Previous issue date: 2007-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The objectives of the present study was the genetical characterizations of Cryptosporidium spp. from different hosts, realize the sequencing an phylogenetic analysis, including the deposit in GenBank of the first Cryptosporidium sequences of animal origin, from Brazil. There were obtained fecal samples, containing Cryptosporidium oocysts from chiken, ducks, quails and Guinea pigs from a public market localized in Rio de Janeiro city, from dairy calfs maintained at a farm localized in the same city and from dogs and cats maintained at a shelter localized in the city of Nova Igua?u. For the analysis was utilized the Nested-PCR of the extracted DNA from 200μl of fecal suspension. For primary identification of Cryptosporidium species was realized RFLP with enzymes SspI and VspI. DNA samples were sequenced and phylogenetic analysis were conducted. There were diagnosed and sequenced C. baileyi infecting two ducks (DQ855339 and DQ885340) and one quail (DQ885335) and C. melagridis infecting one chicken (DQ885341). The sequences obtained form Cryptosporidium infecting Guinea pigs received accession numbers DQ885337 and DQ885338, both sequences were not identified with known Cryptosporidium species due to the great genetic distance between them and those already available at GenBank, suggesting that it may be a new genotype or species. Parasitizing cats was diagnosed C. felis (DQ885336) and in one dog C. canis (DQ885334). One sample of C. parvum of calf origin was sequenced and received accession number DQ885333. During analysis of RFLP pattern of the nested- PCR product from 18Sr DNA was stated that only C. baileyi has a characteristic digestion pattern. Other Cryptosporidium species should be digested by several other enzymes, for a accurate diagnosis. At phylogenetic analysis was found a greater genetic distance between C. felis and C. canis from Brazil when compared to the reference sequences obtained from GenBank. Based on the phylogenetic groupings, a possible new species of Cryptosporidium from Guinea Pigs calls attention for the existence of new species even in common pet animals. As is the case of the Guinea Pig. The sequences obtained in this study are the first Brazilian sequences of C. baileyi, C. meleagridis, C. felis, C. canis and C. parvum deposited in GenBank. / O presente trabalho teve por objetivo caracterizar geneticamente as esp?cies de Cryptosporidium oriundos de v?rios hospedeiros, realizar o seq?enciamento e an?lises filogen?ticas, incluindo o dep?sito das primeiras seq??ncias brasileiras de Cryptosporidium spp. de origem animal no GenBank. Foram obtidas amostras fecais contendo oocistos de Cryptosporidium de pintos, patos, codornas e porquinhos da ?ndia comercializados num mercado municipal da cidade do Rio de Janeiro, de bezerros de uma propriedade voltada ? produ??o leiteira localizada no mesmo munic?pio e de gatos e c?es de um abrigo para animais localizado no munic?pio de Nova Igua?u. Para as an?lises foi utilizado Nested-PCR do DNA extra?do a partir de 200μl de solu??o fecal. Foi realizada RFLP dos produtos obtidos no Nested-PCR, utilizando-se as enzimas SspI e VspI, para uma identifica??o preliminar das esp?cies de Cryptosporidium presentes. As amostras de DNA foram seq?enciadas e an?lises filogen?ticas foram conduzidas. Foram diagnosticados e sequenciados C. baileyi infectando dois patos (DQ855339 e DQ885340) e uma codorna (DQ885335) e C. melagridis infectando um pinto (DQ885341). As seq??ncias dos Porquinhos da ?ndia receberam os n?meros de acesso DQ885337 e DQ885338, sendo que ambas as seq??ncias n?o puderam ser identificadas como esp?cie conhecida de Cryptosporidium, devido ? grande dist?ncia gen?tica entre elas e aquelas j? depositadas no GenBank, sugerindo que se trate de um gen?ptipo ou esp?cie nova. Parasitando os gatos foi diagnosticado C. felis (DQ885336) e em um c?o C. canis (DQ885334). Uma das amostras de C. parvum de bovinos foi seq?enciada, sendo depositada no GenBank sob n?mero de acesso DQ885333. Durante as an?lises dos s?tios de corte enzim?tico dos produtos da Nested-PCR do gen 18Sr DNA, a ?nica esp?cie que realmente possue padr?o de corte caracter?stico ? C. baileyi. As demais esp?cies de Cryptosporidium deveriam ser submetidas ? a??o de outras enzimas, para um diagn?stico acurado. Nas an?lises filogen?ticas foi observada uma dist?ncia gen?tica maior entre C. felis e C. canis isolados no Brasil quando comparados ?s seq??ncias do GenBank. Com base nos dados apresentados pelo agrupamento filogen?tico, uma poss?vel nova esp?cie chama a aten??o ? presen?a de esp?cies desconhecidas de Cryptosporidium, mesmo em animais comuns de estima??o, como ? o caso do Porquinho da ?ndia. Estas s?o as primeiras seq??ncias de C. baileyi, C. meleagridis, C. felis, C. canis e C. parvum do Brasil depositadas no GenBank.