Submitted by Ana Paula Macedo (ensino@ioc.fiocruz.br) on 2013-10-01T00:42:06Z No. of bitstreams: 1 Karla Bitencourth Garcia.pdf: 2108170 bytes, checksum: dc08fd06fd5448e7cada7447c8cb384a (MD5) / Made available in DSpace on 2013-10-01T00:42:06Z (GMT). No. of bitstreams: 1 Karla Bitencourth Garcia.pdf: 2108170 bytes, checksum: dc08fd06fd5448e7cada7447c8cb384a (MD5) Previous issue date: 2013 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / No Brasil, a principal doença em humanos na qual o carrapato atua como vetor é a Febre Maculosa Brasileira (FMB). No Rio de Janeiro (RJ), Amblyomma cajennense é uma das principais espécies vetoras de um dos bioagentes da FMB, e responsável pelo ciclo epidêmico da doença. A baixa especificidade parasitária de A. cajennense, resistência às adversidades climáticas, ampla distribuição geográfica, variação intra-específica, comportamento trioxêno e antropofílico enfatizam o importante papel dessa espécie na transmissão de patógenos para animais, estabelecendo ou mantendo ciclos enzoóticos, epizoóticos e epidêmicos. Apesar do seu impacto na saúde pública, é incipiente o conhecimento sobre os fatores que influenciam seu potencial patogênico e de transmissão de micro-organismos. Os objetivos foram: a) determinar sequências de DNA dos genes mitocondriais Dloop, Citocromo Oxidase II e 12SrDNA de A. cajennense de diferentes regiões do RJ; b) realizar análises filogeográficas e de estrutura de populações das sequências obtidas de cada gene; c) analisar, estatisticamente, se há associação entre a diversidade genética de A. cajennense e os casos de FMB no RJ. Foram processadas 314 amostras de A. cajennense, provenientes de 19 municípios de sete regiões fisiográficas do RJ. Todas as amostras foram submetidas à extração de DNA e amplificação, por PCR, dos três genes. Amostras positivas na PCR foram sequenciadas. As sequências de cada gene foram submetidas a análises filogeográficas e de estrutura de populações. Para verificar a associação entre os haplótipos de A. cajennense, sequências genéticas e a influência sobre o número de casos confirmados de FMB no RJ, foram realizados os cálculos do coeficiente de correlação por postos de Kendall e coeficiente de correlação parcial por postos de Kendall. As análises revelaram que há expressiva diversidade de A. cajennense no RJ, com o foco da diversidade genética nas regiões Serrana e Centro Sul. Entretanto, não há estruturação populacional dessa espécie de acordo com cada região do Estado. Porém, há indícios de estruturação populacional de A. cajennense entre grupos de regiões do RJ. As análises estatísticas revelaram que há associação entre o número de sequências analisadas e diversidade intra-específica. E ainda, quanto maior a diversidade dessa espécie, maior é a probabilidade de ocorrência de casos confirmados de FMB no RJ. / In Brazil, the most important tick-borne disease is Brazilian Spotted Fever (BSF). In Rio de Janeiro (RJ), Amblyomma cajennense is the major vector species of one of BSF bioagents, and responsible for the disease epidemic cycle. The A. cajennense low host specificity, resistance to adverse weather, wide geographic distribution, intraspecific variation, anthropophilic behavior and being a three host tick emphasizes the important role of these species in pathogens transmission to animals, establishing or maintaining enzootic, epizootic and epidemic cycles. Despite its impact on public health, the knowledge about factors that influence its pathogenic potential and transmission of micro-organisms are incipient. The objectives were: a) determine DNA sequences of mitochondrial genes Dloop, Cytochrome Oxidase II and 12SrDNA of A. cajennense from differents regions of RJ; b) perform phylogeographics and population’s structure analysis of sequences obtained for each gene; c) analyze, statistically, if there is an association between A. cajennense genetic diversity and cases of BSF in RJ. 314 samples of A. cajennense, coming from 19 municipalities of seven RJ physiographics regions, were processed. All samples were subjected to DNA extraction and amplification, by PCR, of the three genes. PCR positive samples were sequenced. The sequences of each gene were submitted to phylogeographics and population’s structure analysis. To investigate the association between A. cajennense haplotypes, genetic sequences and the influence on BSF confirmed cases in RJ, were performed calculation of the Kendall posts correlation coefficient and Kendall posts partial correlation coefficient. The analysis revealed that there is substantial diversity of A. cajennense in RJ, with the focus of genetic diversity in South Center and Highland regions. However, there is no population structure of these species according to each region of the State. But, there is evidence of A. cajennense population structure when analyzes are performed considering groups of RJ regions. Statistical analyzes revealed that there is association between the number of sequences analyzed and intraspecific diversity. Also, the greater is the diversity of these species, higher is the probability of BSF confirmed cases occurrence in RJ.

Submitted by FREDY ARVEY RIVERA PÁEZ null (fredy.rivera@ucaldas.edu.co) on 2017-08-25T20:51:36Z No. of bitstreams: 1 Teses Doutorado Fredy Rivera.pdf: 7911822 bytes, checksum: 83cad331d2fa5ca6862f3987ab4e9e66 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-08-29T13:54:41Z (GMT) No. of bitstreams: 1 riverapaez_fa_dr_rcla.pdf: 7911822 bytes, checksum: 83cad331d2fa5ca6862f3987ab4e9e66 (MD5) / Made available in DSpace on 2017-08-29T13:54:41Z (GMT). No. of bitstreams: 1 riverapaez_fa_dr_rcla.pdf: 7911822 bytes, checksum: 83cad331d2fa5ca6862f3987ab4e9e66 (MD5) Previous issue date: 2017-08-11 / Asociación Universitaria Iberoamericana de Postgrado (AIUP) / Os carrapatos de corpo duro (Acari: Ixodidae) contam com 724 espécies descritas, sendo 120 espécies registradas oficialmente na região neotropical. No entanto, na Colômbia o registro e distribuição das espécies da família Ixodidae é escasso e pouco se sabe sobre a interação destes ectoparasitas com as diferentes espécies de Rickettsia na região, deixando uma lacuna em estudos e em conhecimento ixodológico e zoonótico no país. Desta forma, o presente trabalho buscou: a) realizar o levantamento e identificação de espécies de carrapatos duros associados a hospedeiros domésticos em regiões da Colômbia, utilizando caracteres morfológicos externos e marcadores moleculares; b) aferir a existência de caracteres morfohistológicos do sistema reprodutor masculino de carrapatos do gênero Amblyomma e c) verificar a ocorrência de bactérias Rickettsia spp. em carrapatos duros em regiões da Colômbia. Durante o período de agosto de 2014 e maio de 2016, foram coletados 1.745 carrapatos diretamente de hospedeiros domésticos e em ativa alimentação, em 17 municípios de 10 departamentos da Colômbia. Os indivíduos coletados foram identificados com base em sua morfologia externa e preparados para aplicação de técnicas de microscopia, bem como técnicas de análise molecular dos genes ITS2, citocromo oxidase I (COI) e 16S rDNA. Os resultados encontrados permitiram o registro de três gêneros e oito espécies de carrapatos duros nas áreas de estudo, sendo registrada pela primeira vez a espécie Amblyomma mixtum para a Colômbia. Outro registro inédito, no presente estudo foi um caso de ginandromorfismo em A. mixtum, sendo o primeiro caso para a espécie e o primeiro para carrapatos no país. As análises das populações de Rhipicephalus sanguineus sensu lato mostraram que os carrapatos do departamento do Casanare são morfologicamente distintos aos de outras regiões da Colômbia, apesar das análises moleculares evidenciarem tratar-se da mesma linhagem. Embora ainda preliminar, os dados obtidos mostraram também que o sistema reprodutor masculino de carrapatos e suas células germinativas possuem caracteres elegíveis para análises de sistemática de Ixodidae. Confirmou-se também a presença de agentes infecciosos do grupo das rickettsias em espécimes de carrapatos coletados, alertando sobre a importância de medidas governamentais epidemiológicas em alguns dos departamentos colombianos amostrados. / Hard-bodied ticks (Acari: Ixodidae) comprise 724 known species, with 120 species officially registered for the Neotropical region. However, in Colombia, species registry and distribution for the family Ixodidae is scarce, and little is known of the interaction of these ectoparasites with the different Rickettsia species in the region, leading to gaps regarding the ixodological and zoonotic study and knowledge in the country. In this context, this study aimed to: a) establish and identification hard-bodied ticks associated with domestic hosts in various regions of Colombia, using external morphological characters and molecular markers; b) assess the existence of morphohistological characters of the male reproductive system of Amblyomma ticks; and c) verify the occurrence of Rickettsia spp. in hard-bodied ticks in various regions of Colombia. During August of 2014 and May of 2016, 1745 ticks were collected actively feeding on domestic hosts in 17 municipalities of 10 departments of Colombia. The specimens were identified based on their external morphology and prepared for microscopy, as well as for molecular analyses of genes ITS2, cytochrome oxidase I (COI) and 16S rDNA. The results show the presence of three genera and eight hard-bodied tick species in the municipalities studied, including the first register of the species Amblyomma mixtum for Colombia. In addition, we report an unprecedented case of gynandromorphism in A. mixtum, which also constitutes the first report for the species and the first for ticks in the country. A population analysis of Rhipicephalus sanguineus sensu lato showed that specimens of the department of Casanare are morphologically distinct from those of other regions of Colombia, despite the fact that the molecular analyses showed that these belong to the same lineage. Although preliminary, the data obtained also show that the male tick reproductive system and its germ cells present eligible characters for Ixodidae systematics. We also confirmed the presence of infectious agents of the rickettsial group in the tick specimens collected, which alerts the importance of governmental epidemiological measures in some departments of Colombia.

O protozoário Rangelia vitalii, um piroplasma patogênico para cães, foi descrito no inicio do século XX, porém, apenas recentemente, a espécie foi validada por técnicas de biologia molecular. Observações epidemiológicas têm levado a suspeitar que os carrapatos Rhipicephalus sanguineus e Amblyomma aureolatum sejam potenciais vetores de R. vitalii, muito embora, nenhum estudo tenha, até o momento, comprovado o papel de algum carrapato como vetor de R. vitalii. Desta forma, o presente projeto objetiva: 1- Avaliar a transmissão transovariana de R. vitalii em carrapatos das espécies A. aureolatum, R. sanguineus, Amblyomma ovale, Amblyomma tigrinum e Amblyomma cajennense. 2- Avaliar a perpetuação transestadial de R. vitalii em todos os estágios biológicos das espécies citadas acima; 3- Avaliar a capacidade de larvas, ninfas e adultos de A. aureolatum e R. sanguineus em transmitirem o protozoário R. vitalii para cães, durante o parasitismo; 4- Avaliara a presença do agente em canídeos silvestres das espécies Cerdocyon thous e Lycalopex gymnocercus; 5- Avaliar a capacidade de uma fêmea gestante de Canis lupus familiaris em transmitirem de forma vertical o agente; 6- Conhecer as áreas de distribuição da R. vitalii a partir da pesquisa de casos suspeitos; 7- Avaliar as alterações clínicas e hematológicas dos cães infectados, via carrapato ou inoculação. Para tal, cães foram experimentalmente infectados com cepas patogênicas de R. vitalii oriundas do Rio Grande do Sul. Larvas, ninfas e adultos de R. sanguineus, A. aureolatum, A. ovale, A. tigrinum e A. cajennense foram levados a infestar esses cães infectados, e posteriormente, após ecdise ou postura de ovos em incubadora, os estágios biológicos subsequentes foram testados por PCR para presença de DNA de R. viatlii e levados a infestar cães não infectados, a fim de se verificar a transmissão de R. vitalii. Além disso, foi pesquisado a presença de R. vitalii em cães domésticos e canídeos silvestres das regiões Sul e Sudeste. Os resultados obtidos permitiram adquirir uma melhor compreensão da epidemiologia da rangeliose canina, pois além de comprovar a competência vetorial de A. aureolatum para R. vitalii (e a não competência para as demais espécies de carrapatos testadas), ampliou a distribuição geográfica deste agente nas regiões Sul e Sudeste. O estudo também possibilitou relatar uma infecção natural e persistente de um C. thous por R. vitalii. Diferentemente dos cães domésticos, que manifestam severas alterações clínicas e hematológicas decorrentes da infecção por R. vitalii, foi verificado um caso de infecção 8 assintomática em um C. thous, sugerindo um possível papel de reservatório do agente na natureza, uma vez que este canídeo silvestre é o principal hospedeiro silvestre para o carrapato A. aureolatum. A distribuição geográfica dos casos de rangeliose tanto em cães domésticos com em C. thous coincidem com a distribuição geográfica deste vetor. Portanto, esta espécie de carrapato, que já se mostrou capaz de veicular o agente em condições de laboratório, é provavelmente seu vetor em condições naturais. / O protozoan Rangelia vitalii, a pathogenic piroplasmid of dogs, was described in the beginning of the 20th century; however, only recently this piroplasm species was validated through molecular analysis. Epidemiological observations have implicated the ticks Rhipicephalus sanguineus and Amblyomma aureolatum as potential vectors of R. vitalii, although no tick species has been proved to be a competent vector. Therefore, the present study aimed: 1- to evaluate transovarial transmission of R. vitalii in the tick species A. aureolatum, R. sanguineus, Amblyomma ovale, Amblyomma tigrinum and Amblyomma cajennense. 2- to evaluate transstadial perpetuation of R. vitalii among the post embryonic stages of the tick species mentioned above; 3- to evaluate the competence of larvae, nymphs, and adults of A. aureolatum and R. sanguineus to transmit the protozoan R. vitalii to dogs during parasitism; 4- to evaluate natural infection by R. vitalii in the wild canids Cerdocyon thous and Lycalopex gymnocercus; 5- to evaluate vertical transmission of R. vitalii in a pregnant female of Canis lupus familiaris; 6- to expand the distribution area of R. vitalii by searching the agent in canine clinical cases; and, 7- to evaluate clinical and hematological alterations in dogs that were infected via tick parasitism or direct inoculation. In this regard, domestic dogs were experimentally infected with pathogenic strains of R. vitalii from the state of Rio Grande do Sul, southern Brazil. Larvae, nymphs, and adults of R. sanguineus, A. aureolatum, A. ovale, A. tigrinum and A. cajennense were allowed to feed on these infected dogs, and subsequently allowed to molt or egg laying within an incubator. Part of the molted ticks or hatched larvae was tested by PCR for the presence of R. vitalii DNA, and the other part was allowed to feed on susceptible dogs, in order to verify R. vitalii transmission through tick feeding. In addition, natural infection by R. vitalii was tested in domestic dogs and wild canids from Southern and Southeastern regions of Brazil. The results contributed to a better understand of the epidemiology of canine rangeliosis, as the vector competence of R. vitalii by A. aureolatum was demonstrated (at the same time, vector incompetence was shown for the other species tested), and the geographic distribution of R. vitalii was expanded in the Southern and Southeastern regions of Brazil. In addition, natural infection by R. vitalii was verified for the first time in wild canids (C. thous) from different areas. In contrast to 10 domestic dogs, which usually developed severe clinical rangeliosis with marked hematological alterations, an assymptomatic case was observed in a naturally infected C. thous, suggesting a possible reservoir role, since this wild canid is a natural host of A. aureolatum. The geographic distribution of the natural cases of rangeliosis, either on domestic dogs or C. thous coincides with the geographic distribution of A. aureolatum, which should be considered as the main vector of R. vitalii under natural conditions.

Ehrlichia ruminantium, the causative organism of heartwater infections, places severe economic constraint on the livestock industry wherever Amblyomma tick vectors are present. Angora goats are particularly susceptible to this disease and the current live blood vaccine cannot safely be used to protect these animals. An attenuated E. ruminantium (Welgevonden) experimental vaccine has previously shown promising results in Merino sheep and Boer goats. The vaccine was administered by intravenous route (i/v). The general objective of this study was to test the efficacy and safety of the attenuated heartwater vaccine E. ruminantium (Welgevonden) in Angora goats. The specific objectives were, firstly to assess the intra-muscular route of administration of the attenuated vaccine as compared to the standard i/v route and, secondly, to study the haematological changes in Angora goats before, during and after vaccination under controlled conditions at the Onderstepoort Veterinary Institute tick-free stables. A total of 55 Angora goats were used in this trial. They were purchased from an area in South Africa which is known to be Amblyomma-free and heartwater-free. Furthermore, on arrival, the goats were screened for E. ruminantium infection by the immunofluorescent antibody (IFA) test to confirm their disease-free status. The Angora goats were divided into 3 groups: In Group 1, ten were vaccinated by the standard i/v route, in Group 2, 31 received the vaccine by i/m route and 10 served as untreated controls for Group 3. Five of the 10 i/v vaccinated group, 20/31 of the i/m vaccinated and 5 controls were challenged by feeding of known infected adult A hebreaum. The other remaining animals within the three groups were challenged using a known infected blood stabilate administered by the standard i/v route (dose 5xLD50). All animals were challenged 42 days after vaccination. The vaccine did not produce any inflammatory reactions at the site of injection. However, 3/31 (9.7%) of i/m and 7/10 (70%) of i/v vaccinated goats developed febrile reactions starting on Day 11 post-immunisation and were treated. All vaccinated goats were fully protected against either needle i/v or tick challenge, while the control non-vaccinated goats reacted severely to the challenge materials and required oxytetracycline treatment. Despite treatment, two of the unvaccinated goats died from the challenge material. 9 Haematological values (packed cell volume, differential blood cells count) were obtained on blood samples taken from the treatment and control groups at different times during the course of the trial. Wide within group variations as shown by the high standard deviation values were found. As no significant changes were found between vaccinated and control animals, it is likely that the attenuated vaccine does not cause significant clinical haematological changes. This study has demonstrated that the attenuated E. ruminantium (Welgevonden) vaccine is safe in 90.3% and efficacious (100% efficacy) for intramuscular administration in Angora goats. However, further laboratory and on-farms studies are needed in order to establish the lowest effective and safety dose, duration of immunity, and the vaccine’s safety in young and pregnant animals. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Veterinary Tropical Diseases / unrestricted

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-01-23T15:04:32Z No. of bitstreams: 1 texto completo.pdf: 2499744 bytes, checksum: 53d7b61863a6e4086f9d8de02864e860 (MD5) / Made available in DSpace on 2017-01-23T15:04:32Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2499744 bytes, checksum: 53d7b61863a6e4086f9d8de02864e860 (MD5) Previous issue date: 2016-06-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Os carrapatos são artrópodes hematófagos obrigatórios de grande importância médica e veterinária. Estes artrópodes tem desenvolvido adaptações evolutivas que permitem aos mesmos a sobrevivência por longos períodos de tempo sem se alimentarem ou que, após se alimentarem até a ingurgitação, vivam com a presença de altos níveis de substâncias que estimulam o estresse oxidativo. Além disso, apresentam uma anatomia bem conciliada ao seu modo de vida e aquisição de alimentos. Dentre os principais órgãos do carrapato, as glândulas salivares apresentam grande repertório de proteínas, que desempenham papéis essenciais para o seu desenvolvimento e para a manutenção de sua relação com os hospedeiros vertebrados. Essas proteínas também podem facilitar a transmissão de agentes patogênicos aos hospedeiros. Nesse projeto, o sialome do carrapato Amblyomma sculptum foi analisado utilizando cromatografia líquida acoplada com a espectrometria de massas. Essa abordagem combinada permitiu a identificação de 501 proteínas, das quais, aproximadamente, 15% foram classificadas como proteínas secretadas putativas. As classes do grupo de proteínas secretadas putativas que mais se destacaram foram: a) família das proteínas ricas em glicina; b) família das enzimas secretadas e c) ferritinas. As proteínas ricas em glicina podem desenvolver várias funções, dentre elas, a formação do cone de cemento e a resposta a estresse. Proteínas vinculadas com o citoesqueleto, com a maquinaria de modificação proteica e com o transporte intracelular também foram identificadas, as quais foram reunidas no grupo “putative housekeeping proteins”. A diferença na abundância das proteínas entre os gêneros macho e fêmea, encontradas nas glândulas salivares dos carrapatos A. sculptum, foi indicada pela identificação de proteínas em apenas um dos sexos. As proteínas ricas em glicina, do citoesqueleto e chaperonas foram identificadas em maior número em machos do que em fêmeas, sugerindo que esse gênero esteja melhor preparado para a produção de cemento, resposta ao estresse, modificação proteica e transporte intracelular do que as fêmeas. A caracterização funcional das proteínas permitiu uma correlação dentro de um contexto biológico e facilitou um melhor entendimento sobre o sialome de carrapatos A. sculptum. Essa informação pode auxiliar na identificação de novos candidatos a alvos para o controle de carrapatos, no entanto, estudos adicionais devem ser realizados com o objetivo de melhorar o conhecimento das vias de controle das diferentes funções celulares. / Ticks are obligate hematophagous arthropods of great medical and veterinary importance. These arthropods have developed evolutionary adaptations that allow them to survive for long periods of time without feeding or, after feeding until engorgement, live with the presence of high levels of substances that stimulate oxidative stress. In addition, they have an anatomy well reconciled for their way of life and food acquisition. Among the major organs of the tick, the salivary glands contain a large repertoire of proteins, which play essential roles for the tick’s development and for the maintenance of their relationship with their vertebrate hosts. These proteins may also facilitate the transmission of pathogens to the host. In this project, the sialome of the Amblyomma sculptum tick was analyzed using liquid chromatography coupled with mass spectrometry. This combinatorial approach allowed for the identification of 501 proteins, of which approximately 15% were classified as putative secreted proteins. The classes of the group of putative secreted proteins that most stood out were: a) family of glycine-rich proteins; b) family of secreted enzymes; and c) ferritins. Glycine-rich proteins can perform several functions, including cement cone formation and stress response. Proteins linked to the cytoskeleton, the protein modification machinery and intracellular transport were also identified, which were classified in the putative housekeeping proteins group. The difference in the abundance of the proteins between the male and female gender found in the salivary glands of A. sculptum ticks was indicated by the identification of proteins in only one sex. Glycine-rich, cytoskeletal and chaperon proteins were identified in greater numbers in males than in females, suggesting that the male gender is better prepared for cement production, stress response, protein modification and intracellular transport than females. The functional characterization of the proteins allowed a correlation within a biological context and facilitated a better understanding of the sialome of A. sculptum ticks. This information may aid in the identification of novel candidates for tick control; however, additional studies should be carried out in order to better understand the pathways that control with the different cellular functions.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-01-23T15:42:30Z No. of bitstreams: 1 texto completo.pdf: 1904321 bytes, checksum: 732f95d1a780441aa7e91d87b6821bdc (MD5) / Made available in DSpace on 2017-01-23T15:42:30Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1904321 bytes, checksum: 732f95d1a780441aa7e91d87b6821bdc (MD5) Previous issue date: 2016-02-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo deste trabalho foi construir um catálogo de genes para o genoma funcional dos órgãos internos do carrapato A. sculptum, bem como, avaliar os padrões de expressão gênica no intestino médio (MDG) e ovários (OVA) frente à condições de não-infecção (MNI e ONI) e infecção por Rickettsia amblyommii (MI e OI). Como primeiro passo, foi realizada a construção de um transcriptome de referência à partir da montagem de 9 transcriptomas (200 milhões de reads) derivados do intestino médio (3), dos ovários (2) e das glândulas salivares (4). O De novo assembly gerou um total de 460,445 contigs, a partir dos quais foram preditas 27.308 CDS expressas em no transcriptome total do A. sculptum. O mapeamento das reads derivadas de cada órgão contra o transcriptoma de referencia permitiu a identificação de 25,569 CDS expressas nos intestinos (MDG), 21,230 CDS expressas em ovários (OVA), enquanto que 10.697 CDS foram derivados dos transcriptomas de glândula salivar (SG). A anotação do transcriptoma de referência (27.308 CDS) permitiu a identificação de 23,228 CDS relacionados a processos housekeeping, enquanto que 2.177 CDS apresentaram sequência de peptídeos (SignalP) e foram anotadas como proteínas secretads.Um total de 261 CDS foram anotadas como elementos transponíveis, 20 CDS foram relacionadas à transcritos virais, enquanto que 1,622 permaneceram sem anotação, sendo anotadas com Unknown. Entre as 29 classes de genes housekeeping, destacam-se aqueles relacionados com Sinal de transdução (3.158 CDS), seguido de 1887 CDS anotadas com os transportadores e 1.523 CDS relacionados maquinaria de transcrição. A avaliação individualizada dos transcriptomas MDG e OVA no tocante a infecção e não infecção por R. amblyommii (infectados vs não infectado) revelou um padrão oposto entre os dois órgãos, com quase todos os processos celulares am MDG superexpressos em resposta à infecção por riquétsia, enquanto a maioria dos processos celular em ovário (OVA) foram down regulados em resposta à infecção por R. amblyommii. Essa down regulação de processes em ovários de carrapatos infectados por rickettsiae corrobora estudos in vitro com outras especíceis de carraátos infectados com Rickettsia spp. o que concorda com outros estudos in vivo com foco ovipostue de carrapatos infectados. com queda das funções fisiológicas da femeas ingurgitada e queda no rendimento da oviposição. Também foi observado up-regulação de proteínas no intestino, em resposta à infecção, sendo essas relacionadas com processes de infecção rickettsial em ́celulas de mamíferos bem como outros invertebrados, tais como actina, complexo Arp 2/3, domínio WASP, proteína tirosina quinase, clatrinas, intergrinas e ontre outras. Estes resultados sugerem que, de forma semelhante à verificada em células de mamíferos e em outras 17 espécies de carrapatos, estas proteínas estar envolvidas durante o processo de infecção do trato intestinal de A. sculptum. No entanto, a down regulação das principais vias metabólicas, maquinaria de replicação e de síntese proteica em ovários em resposta à infecção por Rickettsia (OI contra ONI) indica que esta batéria se constiui uma carga metabólica e fisiológica para os ovários. Alpem de sugerir possível alvos a sere avaliados na interação rickettsia-carrapato, estes resultados também dão suporte à outros estudos e estratégicas para uma melhor compreensão da biologia deste vetor, bem como para o desenvolvimento de estratégias de combate e controle biológico, como vacinas e acaricidas. / The aim of this work was to construct a gene catalogue for the functional genome of internal organs from A. sculptum, as well as, to evaluate the patterns of gene expression from midguts and ovaries against non-infection and Rickettsia amblyommii infection condition. First, we construct a reference transcriptome for A. sculptum internal organs assembling 9 transcriptomes (200 million reads) derived from midguts (3), ovaries (2) and salivary glands (4). The assemble generated 460,445 contigs, of which we identified 27,308 CDS expressed at whole A. sculptum transcriptome. The mapping of the reads derived from each organ against this CDS reference allowed the identification of 25,569 CDS expressed at midguts (MDG), 21,230 CDS expressed at ovaries (OVA) while 10,697 CDS were derived from sialomes (SG). The annotation of the reference transcriptome (27,308 CDS) allowed the identification of 23,228 CDS related to housekeeping processes, 2,177 CDS related presented signal peptide sequence by SignalP analysis, been annotated as Secreted group; 261 CDS were annotated as transposable elements, 20 CDS were reçated to viral transcripts and 1,622 remained without annotation, been classified as unknown sequences. Among the 29 classes related to housekeeping processes, stand out those related to sinal transduction 3,158 CDS, followed by 1,887 related to transporters and 1,523 CDS related to transcription machinery. The individualized evaluation of the MDG and OVA dataset regarding rickettsial infection (infected vs non-infected) reveals an opposite pattern, with almost all midgut processes more abundant in response to rickettsial infection, while most of all ovarian processes were down regulated in response to rickettsial infection, which agrees to other in vivo studies focusing ovipostue of infected ticks. Regarding MDG datasets, we have observed more abundance of host protein related to rickettsial infection processes at mammals and tick cells, such as actin, Arp 2/3 complex, WASP domain, protein tyrosine kinase, clathrins, intergrins and other. These results suggest that, similarly to verified at mammals cells and other tick specie, these proteins could be play a role during the mechanism of infection of A. sculptum midguts by R. amblyommii. However, the down regulation of metabolic pathways, replication and protein synthesis machinery at ovaries in response to rickettsial infection (OI versus ONI) indicates that R. amblyommii play role as a metabolic and physiological burden for ovaries. These results suggest putative candidates proteins for a rickettsial infection mechanism of A. sculptum midguts and give support for further studies and strategies focused on biology control of the tick, as well, a better understanding about the interface A. sculptum – SFG rickettsiae.

Lumpy skin disease (LSD) is an economically important and debilitating disease of cattle caused by the lumpy skin disease virus (LSDV), a poxvirus in the genus Capripoxvirus. The disease is of economic importance to farmers in endemic regions and is a major constraint to international trade in livestock and their products. It is characterised by fever, enlargement of superficial lymph nodes, loss of weight, inappetence, salivation, lachrymation and formation of eruptive circumscribed skin lesions. The quality of meat and milk are reduced; there is infertility due to reduced sperm quality, abortions and reduced calving rates. The hides are permanently scarred, thereby reducing their quality and trade may be affected following movement restrictions from affected areas. v Lumpy skin disease has the potential to become an emerging disease because of global climate change and changes in patterns of trade in animals and animal products. The disease has become endemic in Africa except in countries like Libya, Algeria, Tunisia and Morocco, where the disease has never been reported. It has also spread to the Middle East where outbreaks were first reported in Israel (1989), Kuwait (1991), Saudi Arabia (1990) , Lebanon (1993), The United Arab Emirates (2000) and Oman (2010). In endemic areas, LSD outbreaks are common in summer. The persistence of LSDV between inter-epidemic periods has not been determined and there is no carrier state reported in either cattle or wild animals. Transmission of the disease has been associated with a high incidence of biting insects such as in wet conditions. The spread of LSD from Egypt to Israel e.g. was associated with movement of the stable fly, Stomoxys calcitrans. The virus has been recovered from S. calcitrans and Biomya fasciata, caught while feeding on infected animals and transmission by insects is suspected to be mechanical, which has been demonstrated in Aedes aegypti mosquitoes. During the 1957 outbreak of LSD in Kenya, affected animals were observed to have high tick infestations, especially of Amblyomma species. In a pilot trial in 2008 at the University of Pretoria (UP), Department of Veterinary Tropical Diseases (DVTD), Amblyomma hebraeum, Rhipicephalus appendiculatus and R. (B) decoloratus ticks were implicated in the transmission of LSDV. The overall objective of this study was to investigate the vector competence of three common sub-Saharan tick species (R. (B) decoloratus, R. appendiculatus and A. hebraeum) and their potential roles in the epidemiology of LSD. This was achieved by testing for persistence of LSDV in ticks and its subsequent transmission to recipient animals following interrupted feeding, transstadial and transovarial development of the ticks. The over-wintering of LSDV was also investigated during transstadial passage in A. hebraeum and transovarial passage in R. (B) decoloratus. During the study, seven cattle were artificially infected with LSDV to serve as source (donors) of infection to ticks. To test for mechanical / intrastadial transmission and persistence in ticks, adult ticks (A. hebraeum and R. appendiculatus) were partially fed on donor animals and then transferred to recipient animals or collected for testing. To test for transstadial transmission/passage, nymphal stages of A. hebraeum and R. appendiculatus were fed on donor animals until they engorged and dropped. Engorged nymphs were incubated to moult to adults. The emergent adults were placed on recipient animals and also tested for the virus. To test for transovarial transmission and passage R. (B) decoloratus (one- host tick) larvae were fed on donor animals until engorged adults. For R. appendiculatus and A. hebraeum (three-host ticks), adults were fed to repletion on the donor animals. Engorged females were collected and incubated to lay eggs and the eggs were allowed to hatch. The emergent larvae were placed to feed on recipient animals to test for transovarial transmission, while larvae were tested for the presence of the virus. Over-wintering of LSDV in ticks was tested by transstadial passage in A. hebraeum and transovarial passage in R. (B) decoloratus under fluctuating reduced temperatures, simulating wintery climatic conditions. Engorged A. hebraeum nymphs and R. (B) decoloratus females were infected by intracoelomic injection. The presence of the virus in LSDV- infected animals was tested by real-time PCR, virus isolation (VI), and the serum neutralisation test (SNT). Tick saliva was tested by real-time PCR and VI while ticks were tested by immunohistochemistry, transmission electron microscopy, VI and real-time PCR. Mechanical/intrastadial and transstadial transmission is reported in A. hebraeum and R. appendiculatus. Transovarial transmission was reported in A. hebraeum, R. appendiculatus and R. (B) decoloratus. The virus was demonstrated in saliva and tick organs of A. hebraeum and R. appendiculatus adults following both mechanical/intrastadial and transstadial persistence. Transovarial passage of LSDV was demonstrated in R. (B) decoloratus, R. appendiculatus and A. hebraeum larvae. The virus also persisted through cold temperature exposure during transstadial passage in A. hebraeum and transovarial passage in R. appendiculatus. This study confirms the vector competency of A. hebraeum, R. appendiculatus and R. (B) decoloratus ticks for LSDV. It also shows the potential for LSDV to over-winter in ticks and demonstrates that LSDV may persist in ticks during inter-epidemic periods. / Thesis (PhD)--University of Pretoria, 2013. / gm2014 / Veterinary Tropical Diseases / unrestricted

Amblyomma maculatum (the Gulf Coast tick), an aggressive, human-biting, Nearctic and Neotropical tick, is the principal vector of Rickettsia parkeri in the United States. This pathogenic spotted fever group Rickettsia species has been identified in 8-52% of questing adult Gulf Coast ticks in the southeastern United States. To our knowledge, R. parkeri has not been reported previously from adult specimens of A. maculatum collected in Kansas or Oklahoma. A total of 216 adult A. maculatum ticks were collected from 18 counties in Kansas and Oklahoma during 2011-2014 and evaluated by molecular methods for evidence of infection with R. parkeri. No infections with this agent were identified; however, 47% of 94 ticks collected from Kansas and 73% of 122 ticks from Oklahoma were infected with ". Candidatus Rickettsia andeanae" a spotted fever group Rickettsia species of undetermined pathogenicity. These preliminary data suggest that " Ca. R. andeanae" is well-adapted to survival in populations of A. maculatum in Kansas and Oklahoma, and that its ubiquity in Gulf Coast ticks in these states may effectively exclude R. parkeri from their shared arthropod host, which could diminish markedly or preclude entirely the occurrence of R. parkeri rickettsiosis in this region of the United States.

No description available.

A Febre Maculosa Brasileira (FMB) é uma doença infecciosa causada por Rickettsia rickettsii, transmitida pelo carrapato-estrela Amblyomma sculptum no interior do estado de São Paulo, onde equinos e capivaras são utilizados como sentinelas para FMB por serem considerados hospedeiros primários deste carrapato. Objetivou-se com este trabalho verificar a ocorrência de anticorpos anti-Rickettsia rickettsii em sentinelas clássicos (equinos) e potenciais (ovinos, bovinos e gambás) para FMB que vivem em diferentes ambientes do Campus \"Luiz de Queiroz\", da Universidade de São Paulo, em Piracicaba-SP. De fevereiro de 2017 a janeiro de 2018 foram amostrados 156 hospedeiros, 48 que habitam áreas urbanas, sem a presença ou trânsito de capivaras (15 equinos e 33 gambás), 60 ovinos que frequentam pastagens sem presença de capivaras, mas adjacentes a matas ciliares ocupadas pelos roedores, e 48 que frequentam pastagens por onde transitam capivaras (31 bovinos e seis equinos) ou matas ciliares ocupadas pelos roedores (11 gambás). Quando possível, a amostragem envolveu a coleta de ectoparasitos. O soro obtido após centrifugação do sangue colhido foi analisado em duas etapas, triagem e titulação, pela reação de imunofluorescência indireta (RIFI). Na triagem todas as amostras foram testadas contra R. rickettsii, e consideradas positivas se apresentassem título de anticorpos >= 64. As amostras positivas foram diluídas na etapa de titulação e testadas contra R. rickettsii, R. parkeri, R. bellii, R. ambyommatis e R. rhipicephali para determinação do título máximo de anticorpos para cada espécie. Das 156 amostras testadas, 15 estavam soropositivas, 1/60 ovinos, 1/31 bovinos, 7/21 equinos e 6/44 gambás; os títulos finais variaram de 64 a 4.096. Foi possível determinar o possível antígeno envolvido em reação homóloga (PAERH) em seis amostras, um equino (R. bellii) e quatro gambás e um ovino (R. rickettsii). Foram registrados os carrapatos Amblyomma ovale, Amblyomma dubitatum, Amblyomma sculptum, Dermacentor nitens e Rhipicephalus microplus, e as pulgas Ctenocephalides felis e Siphonaptera: Rhopalopsyllidae cf. Rhopalopsyllus nos hospedeiros amostrados. Conclui-se que equinos e gambás podem ser utilizados como sentinelas para FMB no Campus \"Luiz de Queiroz\", mas os ovinos não. Não foi possível concluir se os bovinos podem ser utilizados com esta função. R. bellii, ou um microrganismo muito semelhante, ocorre na área urbana do Campus \"Luiz de Queiroz\" e é capaz de infectar cavalos. R. rickettsii, ou um microrganismo muito semelhante, ocorre no Campus \"Luiz de Queiroz\" em áreas de permanência e trânsito de capivaras / Brazilian Spotted Fever (BSF) is an infectious disease caused by Rickettsia rickettsii, transmitted by the tick Amblyomma sculptum outside the metropolitan region in the State of São Paulo, where horses and capybaras are used as sentinels for BSF as they are considered primary hosts for this tick. This survey aimed to verify the occurrence of antibodies against Rickettsia rickettsii in sentinels (horses) and potential sentinels (sheep, cattle and opossums) for BSF that lives in different environments of the \"Luiz de Queiroz\" Campus, University of São Paulo, in Piracicaba, SP, Brazil. A total of 156 host were sampled between February of 2017 and January of 2018; 48 that live in urban areas, with no presence or transit of capybaras (15 horses and 33 opossums), 60 sheep that regularly graze on capybara-free pastures adjacent to a riparian forest occupied by these rodents, and 48 that occupy, cross or regularly graze on pastures (31 cows and six horses) or riparian forests (11 opossums) that are areas used by capybaras. Whenever possible, host sampling included ectoparasites collection. Sera obtained after centrifugation of the collected blood was analyzed in two steps, screening and titration, by the indirect immunofluorescence assay technique (IFA). All samples were tested against R. rickettsii during the screening, and those that presented antibodies titers >= 64 were considered positive. Positive samples were diluted during titration and tested against R. rickettsii, R. parkeri, R. bellii, R. ambyommatis and R. rhipicephali for determination of maximum antibodies titers for each species. Of the 156 tested samples, 15 were seropositive, 1/60 sheep, 1/31 cows, 7/21 horses and 6/44 opossums; antibodies endpoint titers ranged from 64 to 4.096. Possible antigen involved in a homologous reaction (PAIHR) was determined for six samples, one horse (R. bellii), four opossums and one sheep (R. rickettsii). The ticks Amblyomma ovale, Amblyomma dubitatum, Amblyomma sculptum, Dermacentor nitens and Rhipicephalus microplus, and the fleas Ctenocephalides felis and Siphonaptera: Rhopalopsyllidae cf. Rhopalopsyllus were collected from the sampled hosts. Results showed that horses and opossums may be used as sentinels for BSF in the \"Luiz de Queiroz\" Campus, but the sheep do not. It wasn\'t possible to determine if the cows may be used as sentinels. R. bellii, or a very closely related microorganism, occurs in the urban area of the \"Luiz de Queiroz\" Campus and is capable to infect horses. R. rickettsii, or a very closely related microorganism, occurs in capybara-transit areas of the \"Luiz de Queiroz\" Campus