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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.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:tede/803 |
| Date | 27 February 2007 |
| Creators | Huber, Franziska |
| Contributors | Bomfim, Teresa Cristina Bergamo do |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Curso de P?s-Gradua??o em Ci?ncias Veterin?rias, UFRRJ, Brasil, Parasitologia Veterin?ria |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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