Made available in DSpace on 2016-04-28T20:16:24Z (GMT). No. of bitstreams: 1 2007- Anselmo Afonso Golynski.pdf: 441467 bytes, checksum: 4be0e7906cd49bbe4d5b13b8a892680b (MD5) Previous issue date: 2007-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The present study aimed to determine the prevalence of Babesia equi and B. caballi in horses from the northern region of the State of Rio Grande do Sul, examining the maintenance practices and identifying the principal factors involved in transmission and infection rates. There were collected 380 blood samples and tested with ELISA and Indirect immunofluorescence (RIFI) for B. equi and for B. caballi was used competitive ELISA. The sample number was statistically representative for the equine population of the state. Testing positive for B. equi by ELISA and RIFI were 31,6% and 35,8%, respectively, of the samples examined. The prevalence of B. caballi was 0,86%. The concordance between tests was considered high as shown by the Kappa index of 0,86%. There was not observed a statistically significant difference (p<0,05) between the sex, age and breed of the horses. According to the present study the northern region of the state of Rio Grande do Sul can be characterized as an area of enzootic instability for B. equi and B. caballi, offering risks of economic losses due to babesiosis outbreaks, especially in sensible animals introduced from indene areas, horse trade and participating in events (as rodeos, auction sales, and others) or even in animals born in the region. Analyzing the questionnaires, the maintaining systems and the purposes of the equine use were the factors that had influence on the prevalence of equine babesiosis. / O trabalho teve por objetivo determinar a preval?ncia da Babesia equi e B. caballi em eq?inos da regi?o Norte do Estado do Rio Grande do Sul bem como conhecer as pr?ticas de manejo adotadas e identificar os principais fatores que est?o envolvidos na sua transmiss?o e preval?ncia. Coletaram-se 380 amostras de sangue, n?mero estatisticamente representativo para a popula??o de eq?inos em estudo, as quais foram analisadas por meio do teste de ELISA e pela rea??o de imunofluoresc?ncia indireta (RIFI) e para a B. caballi foi utilizado o Elisa competitivo. Do total das amostras, 31,6% e 35,8% foram positivas para B. equi ao teste de ELISA e RIFI respectivamente, sendo que, a preval?ncia de B. caballi foi de apenas 0,86%. A concord?ncia entre os testes foi considerada ?tima, atrav?s do ?ndice Kappa de 0,87. N?o foi constatada nenhuma diferen?a significativa estatisticamente (p<0,05) entre o sexo, ra?as e faixas et?rias dos eq?inos. De acordo com o presente estudo a regi?o Norte do Estado do Rio Grande do Sul ?, portanto caracterizada como uma ?rea de instabilidade enzo?tica para B. equi e B. caballi oferecendo riscos reais de ocorr?ncia e perdas econ?micas causadas por surtos de babesiose causada por B. equi e B. caballi especialmente em animais sens?veis, procedentes de ?reas indenes, com?rcio de animais, participa??o em eventos (esportivos ou recreativos dentro ou fora da Regi?o ou Estado) ou mesmo entre animais nascidos naquela regi?o. Ap?s a an?lise dos question?rios, os sistemas e as finalidades da cria??o foram os ?nicos fatores que influenciaram na preval?ncia das babesioses equinas.

Made available in DSpace on 2016-04-28T20:16:24Z (GMT). No. of bitstreams: 1 2007- Katia Roberta Fernandes.pdf: 2143619 bytes, checksum: 6ff7127f4642317cf1f53d7aceb93f1f (MD5) Previous issue date: 2007-02-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The aim of the present study was to evaluate the natural and experimental infection of B. equi in nymphs and adult Boophilus microplus using optical microscopy and molecular analysis. The experimental infection was observed in nymphs, males and females of B. microplus fed on equines chronically infected with B. equi and in non fed larvae and eggs. For the experiments two equines, of not defined breed, proven to be infected with B. equi were used. The animals were infested with B. microplus larvae of generation F4 obtained from a laboratory maintained population, known to be free of Babesia spp. infection. Daily collections of ticks were realized using as starting point the ecdisys to nymph state. After the collection the hemolymph was examined and the intestine and salivary glands were dissected, smashed on glass slides for microscopy, methanol treated and stained with Giemsa. There were dissected 860 specimens (432 nymphs, 280 females and 148 males). The dissected salivary glands were divided into two parts. The first one was smashed on microscopy glass slides, stained and examined by optical microscopy for morphologic analyses. From the second part was realized DNA extraction and PCR with specific primers for the 18S rRNA. On optical microscopy of nymph hemolymph was possible to be observed the presence of kinetes with claviform aspect characteristic for the genus Babesia. In the salivary glands of B. microplus nymphs the morphology and the sequence of developmental stages of B. equi were observed initiating on day 4 after ecdysis, being possible to see in acinary cells the formation of sporoblasts and ovoid sporozoites measuring 1.5 &#956;m of diameter and elongated sporozoites with 3.8 to 5.2&#956;m of length and 0.8 to 1.5 &#956;m of width. The PCR confirmed the presence of B. equi in DNA samples extracted from salivary glands of nymphs, male and female ticks as well as in larvae and eggs. To evaluate the natural infection were collected nymphs and adult B. microplus from two equines naturally infested by these ticks and naturally infected with B. equi. These horses were originated from Seropedica city in the state of Rio de Janeiro. There were dissected 324 specimens (145 nymphs, 138 females and 41 males). The proceedings with the salivary glands were identical to the previously described for the experimental infection. Of the salivary glands submitted to PCR, 70% showed to be infected with B. equi. Microscopical analysis of the salivary glands revealed the presence of sporoblast stages and the development of elongated sporozoites in acinary glands. The morphologic, morphometric and molecular analysis confirmed the experimental and natural infection of the salivary glands of nymphs and adult B. microplus with B. equi. The results of the present study show the ability of B. equi to develop in this tick species. The detection of B. equi DNA in eggs and larvae also suggests the possibility of transovarian transmission in B. microplus. The results allow to consider the tick B. microplus as a potential biologic vector of B. equi in horses from the studied region. / Este trabalho teve como objetivo avaliar a infec??o natural e experimental de Babesia equi em ninfas e adultos de Boophilus microplus por meio de microscopia ?ptica e an?lise molecular. A infec??o experimental foi observada a partir de ninfas, machos, f?meas, ovos e larvas n?o alimentadas de B. microplus alimentados em equinos com infec??o cr?nica por B. equi. Para a realiza??o do experimento foram utilizados dois eq?inos, mesti?os, com infec??o por B. equi. Os animais foram infestados com larvas de B. microplus de gera??o F4 obtidas de col?nia mantida em laborat?rio e livres de infec??o por Babesia spp. A partir da ecdise para ninfa foram realizadas coletas di?rias. Ap?s a coleta foram realizados os exames de hemolinfa e extra??o do intestino e das gl?ndulas salivares os quais foram macerados em l?minas de vidro para microscopia, fixados em metanol e corados com Giemsa. As gl?ndulas salivares dissecadas foram divididas em duas partes. A primeira parte foi macerada em l?minas de vidro para microscopia, corada com corante Giemsa e observada em microsc?pio ?tico para an?lise morfol?gica. A segunda parte foi realizada a extra??o de DNA, sendo submetida a PCR com primers especif?cos para o gene 18S rRNA para B. equi. Foram dissecados 860 esp?cimes (ninfas= 432; f?meas= 280 e machos= 148). Na microscopia ?ptica foi poss?vel observar nas hemolinfas das ninfas a presen?a de cinetos com aspecto claviforme t?picos do g?nero Babesia. Nas gl?ndulas salivares, a morfologia e a seq??ncia dos est?gios de desenvolvimento de B. equi das ninfa s de B. microplus, foram observadas a partir do 4? dia ap?s ecdise, sendo poss?vel observar nos ?cinos celulares a forma??o de esporoblastos e de esporozo?tas ov?ides medindo 1,5 &#956;m de di?metro e alongados medindo 3,8 a 5,2&#956;m de comprimento e 0,8 a 1,5&#956;m de largura. A rea??o em cadeia de polimerase (PCR) confirmou a presen?a de B. equi em DNA de gl?ndulas salivares extra?das de ninfas, machos, f?meas, assim como dos ovos e larva. A infec??o natural foi observada a partir de ninfas e adultos de B. microplus coletados de dois eq?inos naturalmente infestados e comprovadamente infectados por B. equi, procedentes do munic?pio de Serop?dica, Rio de Janeiro. Foram dissecados 324 esp?cimes (ninfas= 145, f?meas= 138 e machos= 41). O processamento das gl?ndulas salivares dissecadas foi semelhante ao descrito para infec??o experimental. Das gl?ndulas salivares submetidas a PCR, 70% apresentaram resultados positivos para B. equi. As an?lises por microscopia ?ptica das gl?ndulas salivares das ninfas e dos adultos revelaram a presen?a nos ?cinos celulares os est?gios de esporoblastos e o desenvolvimento de esporozo?tas alongados. As an?lises morfol?gicas, morfom?tricas e moleculares confirmaram a infec??o experimental e natural das gl?ndulas salivares de ninfas e adultos de B. microplus por B. equi. Os resultados deste estudo demonstram a capacidade de multiplica??o de B. equi em gl?ndulas salivares de ninfas e adultos de B. microplus. A detec??o de DNA de B. equi em ovos e larvas de B. microplus tamb?m sugere a possibilidade da transmiss?o transovariana nesta esp?cie de carrapato. Estes resultados sugerem que o carrapato B. microplus ? vetor biol?gico de B. equi na regi?o estudada.