Para determinação da ocorrência de transmissão transplacentária da Theileria equi em neonatos eqüinos foram avaliados 50 potros da raça Puro Sangue Lusitano, machos e fêmeas, bem como suas respectivas mães, logo após o parto. Foram colhidas amostras de sangue total, tanto das mães como dos neonatos, entre as primeiras cinco horas pós parto para pesquisa de Theileria equi e Babesia caballi através da técnica de RT-PCR. Utilizou-se o kit de detecção baseado no fluofóro intercalante de DNA SYBERgreen. Um total de 46% das éguas apresentaram resultado positivo para Theileira equi e 54% se mostraram negativas, enquanto que 66% dos potros apresentaram resultados positivos e 34% negativos, sendo que 73,9% dos potros positivos nasceram de mães também positivas. Já para Babesia caballi, 16% das éguas foram positivas e 84% negativas, assim como 2% dos potros foram positivos e 98% negativos. O teste de RT-PCR é bastante sensível e específico, mas pode resultar em falso negativo, apesar de ser eficaz na detecção da Theileria equi e Babesia caballi nos eqüinos. Estes dados permitem concluir que existe a possibilidade de transmissão transplacentária de Theileria equi. / The occurrence of transplacentary transmission of Theileria equi in horses was determined by evaluating 50 young male and female horses of the breed Lusitano Horses as well as their respective mothers. Colts and fillies were evaluated as soon as they were born. Total blood samples were collected from both mother and offspring within the first five hours right after the parturition to analyse Theileria equi and Babesia caballi through the RT-PCR technique. It was used the kit of detection based on DNA SYBERgreen. This study showed us that 46% of the female horses had positive results for Theileira equi and 54% negative results while 66% of the male horses had positive results and 34% of them, negative ones. Moreover, 73.9% of the positive young horses also had their mothers positive. However, for Babesia caballi 16% of the female horses had positive results and 84% negative ones while 2% of the male horses had positive results and 98%, negative ones. The RT-PCR test is very sensitive and specific but it can occur false-negative results although it is efficient in detecting Theileria equi and Babesia caballi in horses. In conclusion, the data show us that there is a possibility of transplacentary transmission of Theileria equi.

La babésiose canine est une maladie parfois mortelle due a des protozoaires des genres Babesia et Theileria transmis par des tiques dont Rhipicephalus sanguineus. Malgré la présence soutenue de R. sanguineus dans le Sud de la France, son rôle dans la transmission de la maladie dans cette région reste incertain. D'autre part, le statut taxonomique de R. sanguineus reste confus et mérite d'être clarifié. Les objectifs de ce travail ont été de caractériser (i) les tiques R. sanguineus s.l. parasitant les chiens dans différentes régions du Sud de la France, (ii) les espèces des genres Babesia/Theileria potentiellement transmises par ces tiques et (iii) l'impact des variations génétiques et/ou des co-infections sur la compétence vectorielle. Au total 140 sangs de chien et 242 tiques R. sanguineus s.l. ont été récoltées et analysées entre 2010 et 2012 par PCR, PCR-RFLP et séquençage. De l'ADN de B. vogeli et B. canis a été détecté dans 13,6% et 12,9% des chiens et dans 10,7% et 1,7% des tiques respectivement avec, parfois, des prévalences significativement différentes entre les régions. Un seul chien a été détecté porteur d'ADN de T. annae. Des comparaisons phylogénétiques et phylogéographiques de séquences de R. sanguineus (ADNr-mt 12S et 16S) et B. vogeli (ARNr 18S) à l'échelle mondiale ont confirmé l'affiliation des tiques de France aux R. sanguineus des régions « tempérées » et suggèrent l'existence d'une coévolution entre le protozoaire et son vecteur. Ce travail apporte de nouvelles connaissances sur la babésiose canine en France et supporte l'existence d'au moins deux populations au sein du groupe sanguineus pouvant présenter des compétences vectorielles différentes / Study on Rhipicephalus sanguineus sensu lato ticks and their potential role in the transmission of canine babesiosis in France: prevalence of parasite infections, vector genetic diversity and epidemiology. Canine babesiosis is a potentially fatal disease caused by protozoa of the genera Babesia and Theileria transmitted by ticks including Rhipicephalus sanguineus. Although R. sanguineus is a major tick species in southern France, its role in the transmission of the disease in this area remain unknown. Moreover, the taxonomic status of R. sanguineus is controversial and needs to be clarified. The aims of this work were to characterize (i) the ticks R. sanguineus s.l. that parasitize dogs in different areas of southern France, (ii) the Babesia/Theileria species potentially transmitted by these ticks in this area and (iii) the impact of genetic variations and/or co-infections on vector competence. A total of 140 dog bloods and 588 R. sanguineus s.l. were collected from 2010 to 2012 from which all dogs bloods and 242 ticks were screened using specific PCR, PCRRFLP and sequencing. B. vogeli and B. canis DNA were evidenced in 13.6% and 12.9% of dogs and in 10.7% and 1.7% of R. sanguineus ticks respectively with sometimes significant prevalence differences between areas. A single case of T. annae infection was detected in a dog. Phylogenetic and phylogeographic analyzes performed at a global scale on 12S and 16S mt-rDNA for ticks and 18S rDNA for B. vogeli confirmed the affiliation of R. sanguineus from France with “temperate” species and suggested the existence of a coevolution process between the pathogen and its vector. This work provides new information on the epidemiology of canine babesiosis in France and supports the existence of at least two populations in the R. sanguineus group in the world with possible different vector competences

Resumen: En esta tesis doctoral se presentan tres modelos matemáticos que describen el comportamiento de dos enfermedades parasitarias con vector de transmisión; de los cuales dos modelos están dedicados a la Toxoplasmosis donde se explora la dinámica de la enfermedad a nivel de la población humana y de gatos domésticos. Los gatos juegan un papel de agentes infecciosos del Toxoplasma gondii. La dinámica cualitativa del modelo es determinada por el umbral básico de reproducción, R0. Si el parámetro R0 < 1, entonces la solución converge al punto de equilibrio libre de la enfermedad. Por otro lado, si R0 > 1, la convergencia es al punto de equilibrio endémico. Las simulaciones numéricas ilustran diferentes dinámicas en función del parámetro umbral R0 y muestra la importancia de este parámetro en el sector salud. Finalmente la Babesiosis bovina se modela a partir de cinco ecuaciones diferenciales ordinarias, que permiten explicar la influencia de los parámetros epidemiológicos en la evolución de la enfermedad. Los estados estacionarios del sistema y el número básico de reproducción R0 son determinados. La existencia del punto endémico y libre de enfermedad se expone, puntos que dependen del R0, ratificando la importancia del parámetro umbral en la salud publica. Objetivo: Construir modelos matemáticos epidemiológicos aplicados a enfermedades parasitarias (Toxoplasmosis y Babesiosis) con vector de transmisión. Metodología: Para la construcción de los modelos matemáticos epidemiológicos es necesario representar la enfermedad a partir de modelos de flujo, permitiendo ver la dinámica de la población entre los diferentes estadíos de la enfermedad, dichos movimientos son analizados a partir de sistemas dinámicos, análisis matemático y métodos numéricos; con estas herramientas es posible hacer un estudio detallado del modelo, permitiendo calcular parámetros umbrales que dominan la dinámica de la enfermedad y a su vez simular escenarios reales e hipotéticos. / Aranda Lozano, DF. (2011). Modeling of parasitic diseases with vector of transmission: toxoplasmosis and babesiosis bovine [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/11539 / Palancia

Tick-borne diseases are caused by a group of pathogenic microorganisms which are transmitted between animal and human population by vector which is most frequently tick. Evaluation of danger and perils of these diseases is important due to prevention and monitoring of current situation development. The most common tick-borne diseases in the Czech Republic are borreliosis, tick-borne encephalitis and ehrlichiosis. Rarely can we encounter bartonellosis, babesiosis, rickettsiosis and tularemia. All these diseases can have serious consequences and in critical cases they can result in death of the infected person. One of the basic and the most important of preventive measures which lower the risk of the infection by any tick-borne disease is prevention of tick encounter and eventually vaccination. Due to continual global warming we can expect that in near future ticks will spread even to locations which have been so far not suitable for their development. We can expect that we will even more frequently encounter tick-borne diseases - including diseases which were formerly not found in our territory or were very rare. As a part of health education it will be needed more to get known basic information of tick-borne diseases and their prevention. Different educational centres are an ideal place for...

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2018-05-09T13:59:39Z No. of bitstreams: 1 2010 - Marcos Pinheiro Franque.pdf: 14895648 bytes, checksum: 175b19f715efd4282c83347a6d0ce674 (MD5) / Made available in DSpace on 2018-05-09T13:59:39Z (GMT). No. of bitstreams: 1 2010 - Marcos Pinheiro Franque.pdf: 14895648 bytes, checksum: 175b19f715efd4282c83347a6d0ce674 (MD5) Previous issue date: 2010-03-19 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico, CNPq, Brasil. / This work describes the dairy farms profile and of its owners, as well as the epidemiological condition of the herd to Babesia bovis, Babesia bigemina and Anaplasma marginale infection in the south mesorregi?o Espirito-santense, with base in a serologic study. Owners or manager of 38 dairy farms were interviewed with a semi-structured questionnaire and the data analyzed by descriptive statistics. In 97% of the dairy farms, the herd was formed by animals with different degrees of blood girolando. Of all producers, 61% were exclusively dairy farms owners and 34% were also coffee producer. The extensive creation system is used in 61% of dairy farms. The dairy farms management, in 89% and 11% of the cases were accomplished by the owners and administrators, respectively. The serologic study was accomplished using indirect ELISA of 756 samples of bovine serum, 380 of dairy cows and 376 of calves in breast feeding phase. The mesorregi?o studied was classified in situation of stability for B. bovis (98,4%), B. bigemina (96,7%) and A. marginale (96,6%) to dairy cows. In relation to the calves was observed prevalence of 75,5% to Babesia bovis, 78,5% to B. bigemina and 69,2% to A. marginale. In general, the results demonstrated that is possible to increase the production and productivity of these dairy farms, but is necessary the owners adapts to the current competitive productive milk system, mainly about the managerial aspect. In the south mesorregi?o Esp?rito-santense the risk of occurrence of outbreaks of babesiosis or anaplasmosis in adult animals is low. However, preventive measured should be adopted for the calves, mainly for those with less to three months of age. / Este trabalho descreve o perfil de propriedades leiteiras e dos produtores, bem como a situa??o epidemiol?gica do rebanho da mesorregi?o sul Esp?rito-santense em rela??o ? Babesia bovis, Babesia bigemina e Anaplasma marginale, com base em um estudo sorol?gico. Propriet?rios ou administradores de 38 estabelecimentos produtores foram entrevistados com uso de um question?rio semi-estruturado e os dados analisados atrav?s de estat?stica descritiva. Em 97% das propriedades visitadas, o rebanho leiteiro era formado por animais com diferentes graus de sangue girolando. Dentre os produtores, 61% se dedicavam exclusivamente a produ??o leiteira e 34% tamb?m se dedicavam ? cafeicultura. O sistema de cria??o extensiva foi utilizado em 61% das propriedades. Foi observado que o gerenciamento, em 89% e 11% dos casos era realizado pelos propriet?rios e administradores, respectivamente. O estudo sorol?gico foi realizado utilizando o m?todo de ELISA indireto a partir de 756 amostras de soro bovino, dos quais 380 de vacas em lacta??o e 376 de bovinos jovens em fase de aleitamento. Para vacas em lacta??o, a mesorregi?o foi classificada em situa??o de estabilidade para B. bovis (98,4%), B. bigemina (96,7%) e A. marginale (96,6%). Em rela??o aos bovinos jovens foi observada preval?ncia de 75,5% para Babesia bovis, 78,5% para B. bigemina e 69,2% para A. marginale. Os resultados demonstraram que, em geral, as propriedades possuem potencial para aumentar sua produ??o e produtividade, mas os produtores precisam se adequar ao competitivo sistema de produ??o de leite, principalmente no aspecto gerencial. Na mesorregi?o sul Esp?rito-santense o risco de ocorr?ncia de surtos das doen?as do Complexo Tristeza Parasit?ria Bovina ? baixo para os animais adultos. Contudo, medidas preventivas devem ser adotadas para os bezerros, principalmente para aqueles com menos de tr?s meses de idade.

Tikslas: išanalizuoti įvairių veiksnių įtaką šunų babeziozės pasireiškimui, plitimui, atsižvelgiant į šuns lytį, amžių ir veislę, naudojant literatūros šaltinius bei asmeninius tyrimus. Duomenys buvo surinkti iš " Jakovo veterinarijos centras", smulkių gyvūnų klinikos "Pas Filą " ir kraujo tyrimai buvo renkami ir atliekami Veterinarijos ir diagnostikos centre. Iš viso surinkome 1195 ligos atvejų, atsižvelgiant į lytį, amžių bei veislę. Anamnezė buvo surinkta iš 117 šunų, kraujo mėginiai buvo imami iš 31 sergančio šuns, kraujo morfologiniams tyrimams. Išanalizavus esamus duomenis, pastebėjome, kad dažniausiai babezioze serga tam tikros šunų veislės: mišrūnai, vokiečių aviganiai, labradoro retriveriai ir sibiro haskiai. Atsižvelgiant į lytį, tai daugiausiai babezioze sirgo patinai ir žymiai rečiau patelės. Babezeliozė dažniausiai buvo diagnozuojama jauniems šunims, 1 iki 2 metų, bet sunkiausia klinika buvo pasireiškusi šuniukams iki 1 metų. Dažniausiai pasireiškianti klinikinė simptomatika: apatija, anoreksija, kliniškai išreikšta anemija, pakitusi kūno temperatūra, tamsus šlapimas, vėmimas, NS sutrikimai, viduriavimas, kliniškai išreikšta gelta. Analizuojant morfologinius kraujo rodiklių pokyčius, nustatėmė, kad visais atvejais buvo pasireiškusi trombocitopenija, sumažėjęs trombokrito kiekis. Pastebėjome, kad ligai būdingas sezoniškumas. Stipriausias babeziozės bumas pasireiškia pavasarį – balandžio, gegužės, birželio mėnesiais ir kiek silpnesnis – rudenį (rugsėjį, spalį). ... [toliau žr. visą tekstą] / Goal: to find out a variety of factors influence the occurrence of canine babesiosis, development, according to dog sex, age and breed, using known literature and personal research. The data were collected from „Jakovo veterinary center“, small animal clinic „pas Filą“ and the blood tests were collected and conducted at Veterinaty and Diagnostic center. We collected 1195 disease cases according to sex, age and breed. Anamnesis were collected from 117 dogs, morphological studies of blood – 31. According to the studies, we found out, that there are certain breeds are more susceptible to babesiosis: mongrels, german shepherds, labrador retrievers and siberian huskies. Also found, that babesiosis more prone males than females. Dog babesiosis were usually diagnosed in young dogs, 1 to 2 years, but the hardest clinic gets puppies up to 1 year. The most frequent clinical manifestations in patients with babesiosis : apathy, anorexia, symptomatic anemia, change in body temperature, dark urine, vomiting, NS disorders, diarrhea, clinical jaundice. Morphological characteristics of the blood we found that in all cases, is characterized by thrombocytopenia, a decrease thrombocyte. The seasonal variation in the number of cases has been reported at all Veterinary clinics, with a higher incidence in the spring months (April, May) and peaking in autumn (September, November).

Background: Canine babesiosis is a tick-borne disease causing detrimental health effects on the domestic dogs with huge economic impact on the owners. The most complicated form of canine babesiosis is caused by a pathogenic Babesia rossi parasite. Canine babesiosis induced by B. rossi still remains the cause of mortality and morbidity in South African dogs, yet, the transcriptomic and genomic information of this parasite species is still not available. The transcriptomic and genomic information is essential in the disease development and processes for the design of effective disease control strategies. Consequently, our understanding of the mechanisms underlying the pathogenesis of the different genotypes of B. rossi remains limited. A previous study suggested a relationship between the parasite genotype and the disease phenotype. To date, thirteen B. rossi genotypes have been identified and associated with diverse clinical signs in their hosts. Hence the aim of this study was to sequence RNA from samples representing B. rossi genotypes, 19, 29 and 31, in order to have insight on the overall transcriptome of this parasite and to establish if there would be significant differences among the genotypes. Methodology: To screen for B. rossi positive samples, total DNA was extracted from 20 blood samples collected from sick domestic dogs presented at the Onderstepoort Veterinary Academic Hospital (OVAH). Babesia rossi infections were confirmed using the PCR-Reverse Line Blot (RLB) hybridization assay. Further confirmation of infection status was done by amplification of the B. rossi Erythrocyte Membrane Antigen 1 (BrEMA1) gene in all the DNA samples using qualitative PCR (qPCR), followed by sequencing of PCR products. Subsequently, total RNA was extracted from the 20 B. rossi-infected blood samples collected from the same dogs in which DNA was extracted. Three samples representing B. rossi genotypes 19, 29 and 31 were selected for transcriptome analysis. RNA sequencing was performed using the Illumina HiSeq 2000 to allow transcriptome analysis. De novo assembly was performed independently for all three transcriptomes using the Trinity software. The unigenes generated from specific transcriptome assemblies were subjected to global functional annotation using Blast2GO version 2.8.0 software, followed by KEGG database for annotation of biological pathways, and DAVID version 6.7, for COG classification to predict and classify their functions. Results: The sample representing B. rossi genotype 31 was excluded in the transcriptome analysis due to low RNA mass, which usually compromises the quality of the library used in RNA sequencing.Thus, a total of 26 747 238 and 25 709 627 paired-end reads were obtained from B. rossi genotypes 19 and 29, respectively. De novo transcriptome assembly produced a total of 3019 unigenes, with an average length of 419 bp and N50 of 362 bp in B. rossi genotype 19, and 2727 unigenes with an average length of 441 bp and N50 of 362 in B. rossi genotype 29. A total of 1193 unigenes were common between B. rossi genotype 19 and 29, while 1828 unigenes were exclusively detected in B. rossi genotype 19; and 1534 were specific to B. rossi genotype 29. Between the two B. rossi genotypes, a total of 4553 unigenes were obtained, representing the overall B. rossi transcriptome. From the overall transcriptome, 12.3% (n=558) of the unigenes could be annotated with 53 different gene ontology (GO) functional categories. About 34% (n=1550) of the unigenes represented in the overall transcriptome mapped to 237 KEGG pathways and only 2.5% (114) could be annotated in the COG database. Conclusion: Although, there were no striking differences in the transcriptomes of B. rossi genotypes 19 and 29, this study presents the first transcriptomic resource for B. rossi, which will highly contribute to our genetic understanding of B. rossi and provide a platform for future gene expression studies. Hypothetical proteins identified in this study will require further characterization as they may have a critical role in the biology and pathogenicity of B. rossi parasite. / Life and Consumer Sciences / M. Sc. (Life Sciences)

Made available in DSpace on 2014-07-29T15:13:55Z (GMT). No. of bitstreams: 1 TESE SABRINA CASTILHO DUARTE.pdf: 306533 bytes, checksum: 4d48881b72114026737e6c40bb7b94fb (MD5) Previous issue date: 2010-03-31 / In the genera Babesia, Ehrlichia and are positioned Hepatozoon species of parasites responsible for babesiosis, ehrlichiosis and hepatozoonosis respectively. These organisms have in common the fact that they are transmitted by tick vectors in dogs infected and cause general symptoms such as fever, anemia and jaundice accompanied by intracellular infections. In the state of Goias had not work with phylogenetic approach for these blood parasites. The overall objective of this study was molecular analysis of isolates of Babesia, Hepatozoon and Ehrlichia obtained from symptomatic dogs in Goiânia, Goiás State, thus confirming that the species and subspecies involved in infections. For this, DNA extraction was carried out of samples, followed by carrying out PCR with generic primers. After the PCR was obtained fragments of the 18S rRNA region for samples of Babesia and Hepatozoon and 16S rRNA for the detection of species of the genus Ehrlichia. PCR products obtained were purified and used for sequencing. We sequenced 35 samples of Babesia spp. and of these 17 were used for phylogenetic studies. Two samples of Hepatozoon spp. 17 samples were sequenced and Ehrlichia spp which only five were used for analysis. Analyses performed with the identity of sequenced samples allowed the identification of B. vogeli canis, Hepatozoon canis and Ehrlichia canis. When confronted with the species and subspecies of reference from other regions of Brazil and the world showed close molecular similarity, which demonstrates the low variability between different samples from different geographic regions. For the analysis using the program MEGA4 and subsequent construction of a phylogenetic tree samples in this study formed their own groups always association with the reference samples for the respective species. Thus it was concluded that samples of B. c. vogeli H. canis and E. canis from dogs in the city of Goiania GO show close similarity with isolates from other regions of the world / Nos gêneros Babesia, Ehrlichia e Hepatozoon são posicionadas as espécies de parasitos responsáveis pela babesiose, ehrlichiose e hepatozoonose respectivamente. Estes microrganismos têm em comum o fato de serem transmitidos por carrapatos vetores e causarem nos cães infectados sinais gerais como febre, anemia e icterícia acompanhados de infecções intracelulares. No estado de Goiás não haviam trabalhos com abordagem filogenética relativos a estes hemoparasitos. O objetivo geral do presente trabalho foi realizar análise molecular de isolados de Babesia, Ehrlichia e Hepatozoon obtidos de cães sintomáticos na cidade de Goiânia, Estado de Goiás, verificando assim qual a espécie e subespécie envolvida nas infecções. Para isto, foi realizado extração de DNA das amostras, seguido da realização de PCR com oligonucleotídeos genéricos. Após a PCR obteve-se fragmentos da região do 18S rRNA para as amostras de Babesia e Hepatozoon e do 16S rRNA para a pesquisa de espécies do gênero Ehrlichia. Os produtos de PCR obtidos foram purificados e utilizados para o sequenciamento. Foram seqüenciadas 35 amostras de Babesia spp. e destas 17 foram utilizadas para os estudos filogenéticos. Duas amostras de Hepatozoon spp. foram seqüenciadas e 17 amostras de Ehrlichia spp sendo que apenas cinco foram utilizadas para as análises. As análises de identidade realizadas com as amostras seqüenciadas permitiram a identificação de B. canis vogeli, Hepatozoon canis e Ehrlichia canis. Quando confrontadas com as espécies e subespécies de referência de outras regiões do Brasil e do mundo apresentaram estreita similaridade molecular, o que demonstra a baixa variabilidade entre as diferentes amostras de diferentes regiões geográficas. Pela análise feita utilizando-se o programa MEGA4 e subsequente construção de árvores filogenéticas as amostras deste estudo formaram grupos próprios sempre em associação com as amostras de referência para as respectivas espécies. Dessa forma foi possível concluir que as amostras de B. c. vogeli, H. canis e E. canis provenientes de cães da cidade de Goiânia-GO apresentam estreita similaridade com isolados de outras regiões do mundo

Made available in DSpace on 2016-08-16T18:18:41Z (GMT). No. of bitstreams: 1 Tese Andressa Almeida Santana.pdf: 1466363 bytes, checksum: ae0eefc75fb09cd9f96594f4e5656ed5 (MD5) Previous issue date: 2011-07-29 / FUNDAÇÃO DE AMPARO À PESQUISA E AO DESENVOLVIMENTO CIENTIFICO E TECNOLÓGICO DO MARANHÃO / Canine babesiosis and canine visceral leishmaniasis (CVL) are vector borne diseases, where dogs exert a play as reservoir or source for arthropods responsible by the transmission of these protozoosis. Babesia canis vogeli, is transmited by tick Rhipicephalus sanguineus while Leishmania infantum (sin. Leishmania chagasi) is transmited by sand fly (Lutzomyia longipalpis). The results showed that CVL remains endemic in São Luís Municipality. Despite that the coinfection between Leishmania and Babesia was low considering that both diseases are endemic in this tropical area. Beside that was observed that Yorkshire terrier presented higher predisposition to acquire the infection by B. canis vogeli. A remarkable result was the occurrence of ocular lesions associated to L. infantum infection. / A babesiose canina e a leishmaniose visceral canina são doenças transmitidas por vetores, sendo os cães competentes reservatórios e fonte alimentar dos artrópodes envolvidos. Babesia canis vogeli, é um parasita intraeritrocitário transmitido pelo carrapato Rhipicephalus sanguineus. Leishmania infantum (sin. Leishmania chagasi) transmitidos a mamíferos pela picada dos flebotomíneos (Lutzomyia longipalpis), infectando macrófagos do Sistema Fagocítico Mononuclear do hospedeiro. O presente trabalho é divido em capítulos e os resultados encontrados mostraram que a leishmaniose visceral canina ainda é endêmica no município de São Luís, e que apesar disso a taxa de coinfecção com Babesia foi baixa. Também foi observado que a raça Yorkshire terrier, dentre as raças estudadas, apresentou maior predisposição para contrair a infecção por B. canis vogeli. Outro resultado significativo foi a ocorrência de lesões oculares associadas à infecção por L. infantum.

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.