Uninfected larvae of Ixodes dammini, lxodes cookei, and Amblyomma americanum were fed on hamsters that had been injected intraperitoneally with a 0.5.ml sample of Borrelia burgdorferl (2.5 X 107 spirochetes per ml) 21 days earlier. A total of 108 of these larvae comprised of 36 1. dammini, 36 i. cookei, and 36 A. americanum were aseptically dissected and examined by darkfield and immunofluorescent microscopy for the presence of B. burgdorferl within 48 hours of feeding on the B. burgdorferi infected hamsters. The removal and examination of the midgut diverticula revealed that 32/36 (88.9%) of the l. dammini larvae contained B. burgdorferl. Only 5/36 (13.9%) of the l. cookei larvae and 7/36 of the A. americanum larvae harbored spirochetes in their midgut diverticula.A portion of the nymphs that molted from the above larvae were also dissected and examined by darkfleld and indirect immunofluorescent techniques. Borrelia burgdorferi were observed in the midgut diverticula of 94/107 (87.8%) of the l. dammini nymphs. None of the 30 (0%) l. cookei nymphs examined were found positive for spirochetes and only 1/60 (1.7%) of the A. americanum nymphs was found positive for B. burgdorrerl.A total of 83 lL dammini, 53 A. americanum, and 161. cookei nymphs reared from larvae that fed to repletion on hamsters infected with B. burgdorrerl were allowed to feed on uninfected hamsters to assess transmission of B. burgdorrerl. Transmission was demonstrated only by the l. dammlnl nymphs. The findings of this study suggest that it is extremely unlikely that l. cookei can serve as a vector for B. burgdorrerl, but do not rule out completely the possibility that A. americanum may be able to maintain B, burgdorrerl infections transstadially and, under certain conditions, transmit the organisms to vertebrate hosts. / Department of Physiology and Health Science

Uninfected nymphs of Ixodes scapularis and Amblyomma americanum were fed on hamsters that had been injected intraperitoneally with a 0.5 ml sample of Borrelia burgdorferi (2.5 X 10' spirochetes per ml) 30 days earlier. All nymphs fed to repletion and were then housed during the molting process. In order to assess their ability to transmit the spirochetes, the resulting l. scapularis and A. americanum adults were allowed to feed on uninfected rabbits.Dissection of the adult l. scapularis ticks revealed 9/12 (75%) to harbor motile spirochetes, identified as B. burgdorferi by darkfield microscopy, isolation in BSK II medium, and indirect immunofluorescent antibody staining with the monoclonal antibody H5332. Transmission was successful to one of two New Zealand White rabbits by these infected ticks.Dissection of the adult A. americanum ticks revealed 0/150 (0%) to harbor spirochetes. Transmission to each of three rabbits was unsuccessful. However, 5/90 (5.6%) cultures of midgut material from these same ticks, harbored non-motile spirochete-like bodies. The identity of these "spirochetes" is unknown. / Department of Biology

In 1994 and 1995, seven cases of Human monocytic ehrlichiosis were reported in Indiana; six cf these were from southern counties. To test whether Ehrlichia cha ffeensis is present in native ticks, adult ticks representing two species, Dermacentor variabilis and Amblyomma americanum were collected in southern Indiana during the first week of May, 1995 and tested by PCR. A total of 510 D. variabilis ticks were collected and placed into 102 pools (5 ticks per pool). A total of 430 adult A. americanum ticks were collected and placed into 88 pools of 5 ticks or less.D. variabilis ticks were used to optimize the isolation of PCR amplifiable DNA and determine the minimum number of bacterial cells detected. A modified CTAB-phenol method permitted the detection of as few as 100 bacterial cells. The optimal amount of isolated DNA for a PCR amplification was determined to be 2.7 pg of total nucleic acid.The 88 pools of adult A. americanum were subjected to DNA isolation, PCR amplification, and Southern analysis to determine the presence of E. chaffeensis bacteria. Using the 16S rRNA gene from E. chaffeensis, with species specific primers Hut and HE3. Of the 88 pools (430 ticks; 21 were determined to be positive for the presence E. chaffeensis bacteria. Thus, minimal field infection rate for adult A. americanum ticks in Southern Indiana was 4.88%. This calculation is based upon the assumption that at least one tick was positive in each positive pool. / Department of Biology

Thesis (Ph. D.)--Ohio State University, 2008.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-06-22T17:13:14Z No. of bitstreams: 1 texto completo.pdf: 1578494 bytes, checksum: d7fa5a752b4c56f8611d2e504d7afa84 (MD5) / Made available in DSpace on 2017-06-22T17:13:14Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1578494 bytes, checksum: d7fa5a752b4c56f8611d2e504d7afa84 (MD5) Previous issue date: 2017-02-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Os carrapatos são artrópodes parasitas que se alimentam obrigatoriamente de sangue, sendo capazes de parasitar diversos hospedeiros vertebrados. O intestino dos carrapatos machos e fêmeas possui diferenças morfológicas, tendo como principal função a digestão intracelular dos componentes do sangue obtidos durante a alimentação. Este órgão possui várias proteínas relacionadas com o desenvolvimento, metabolismo e mecanismo de digestão da hemoglobina. Neste trabalho, as proteínas do intestino de carrapatos Amblyomma sculptum, machos e fêmeas foram analisadas com o auxílio da cromatografia líquida acoplada com a espectrometria de massas. O algoritmo PEAKS identificou um maior número de proteínas do intestino nas estratégias fracionadas e não fracionadas em relação aos algoritmos MASCOT e SCAFFOLD, tanto para o banco de dados Chelicerata quanto para o banco de dados Oryctolagus, utilizando os analisadores Ion Trap e QTOF. Pelo KOG, as classes das proteínas analisadas no intestino dos carrapatos machos e fêmeas que se destacaram estão envolvidas na dinâmica e estrutura da cromatina, tradução, biogênese e estrutura ribossomal, modificação pós-traducional, turnover proteico, chaperonas, citoesqueleto, conversão e produção de energia, metabolismo e transporte de carboidratos, lipídeos e íons inorgânicos. Na identificação das proteínas do hospedeiro, a hemoglobina, componente de maior volume no sangue, foi encontrada em ambos os sexos. As informações obtidas neste trabalho podem fornecer uma base sobre a fisiologia do intestino de machos e fêmeas A. sculptum e auxiliar na escolha de proteínas como alvos moleculares potenciais, visando avanço nas estratégias de controle aplicadas para estes artrópodes. / Ticks are parasitic arthropods that feed necessarily of blood, being able of parasitizing various vertebrate hosts. The midgut of males and females ticks has morphological differences, having as main function the intracellular digestion of blood components obtained during feeding. This organ has several proteins related to the development, metabolism, and mechanism of hemoglobin digestion. In this work, the midgut proteins of Amblyomma sculptum ticks, males and females were analyzed with aid of liquid chromatography coupled with mass spectrometry. The PEAKS algorithm identified a larger number of proteins in fractional and non-fractionated strategies in relation to algorithms MASCOT and SCAFFOLD for both Chelicerata and Oryctolagus databases, using Ion Trap and QTOF analyzers. By KOG, the classes of proteins analyzed in the midgut of the males and females ticks are involved in the chromatin structure and dynamics, translation, ribosomal structure and biogenesis, post- translational modification, protein turnover and chaperones, cytoskeleton, energy production and conversion, metabolism and transport of carbohydrates, lipids and inorganic ions. In the identification of the host proteins, the hemoglobin, greater blood volume component, was found in both genders. The informations obtained in this work can provide a basis about the midgut phisiology of males and females A. sculptum and assist in the selection of proteins as potencial molecular targets, aiming advance in the control strategies applied to these arthropods.

Submitted by Nathália Faria da Silva (nathaliafsilva.ufv@gmail.com) on 2017-10-25T17:11:15Z No. of bitstreams: 1 texto completo.pdf: 1261921 bytes, checksum: 067fce7911dd12871ca7da328ba82529 (MD5) / Made available in DSpace on 2017-10-25T17:11:15Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1261921 bytes, checksum: 067fce7911dd12871ca7da328ba82529 (MD5) Previous issue date: 2017-06-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O carrapato Amblyomma sculptum pertence ao complexo Amblyomma cajennnense, taxon mais importante na transmissão e como hospedeiro da bactéria Rickettsia rickettsii, principal agente etiológico da Febre Maculosa Brasileira e da Febre Maculosa das Montanhas Rochosas nos EUA. Como membro deste complexo de espécies, o carrapato A. sculptum é atualmente muito importante sob o aspecto médico veterinário atuando como vetor deste agente zoonótico no Brasil. Seu papel no ciclo biológico da riquétsia, bem como a relação Rickettsia-carrapato, precisam ser bem compreendidos pois estudos recentes encontraram esta espécie de bactéria no carrapato A. sculptum. Como parte desta relação, as funções de RNAs não codificantes de cadeias longas (lncRNA) são desconhecidas e precisam ser descobertas. Os lncRNAs estão envolvidos em uma infinidade de atividades celulares tais como transcrição, silenciamento gênico e abertura cromossômica, por exemplo. Dessa forma, apresentamos aqui uma abordagem in silico da modulação de lncRNAs pela Rickettsia amblyommii em carrapatos da espécie A. sculptum. Tomando como dados trabalhos publicados anteriormente nós identificamos e avaliamos possíveis lncRNAs para expressão diferencial em intestinos e ovários desta espécie de carrapato. Dois montadores de sequencias foram testados, Trinity e CLC Genomics, para construção de contigs e um filtro a partir de um pipeline caseiro para selecionar apenas sequências com características de lncRNA. Um total de 31.888 e 23.733 contigs foram montados pelo Trinity e CLC Genomics, respectivamente. Nós anotamos mais de 500 sequências possíveis de lncRNA contra os bancos de dados RefSeq RNA, RNA Central e NONCODE. Nossos resultados sugerem que a R. amblyommii está induzindo expressão diferencial de lncRNAs nos tecidos dos intestinos e ovários. Nosso trabalho contribui para o aumento do banco de dados de lncRNA de carrapatos bem como fornecer idéias iniciais de quais sequências de lncRNA estão envolvidas na relação Rickettsia- carrapato / The Amblyomma sculptum tick belongs to Amblyomma cajennnense complex, the most important taxon in transmission and host for Rickettsia rickettsii bacteria, the main etiologic agent of Brazilian Spotted Fever in Brazil and Rocky Mountain Spotted Fever in USA. As a member of this species complex, A. sculptum tick is actually very important under human and veterinary subject acting as vector for this zoonotic agent in Brazil. Its role in biological cycle of rickettsia strains, as well Rickettsia-tick interactions, still need to be better understood since recent studies found these strains infecting A. sculptum ticks. As putative modulators of these pathogen-host interaction, the roles of long non-coding RNAs (lncRNA) of ticks are unknown and need to be discovered and characterized. The lncRNAs are involving in modulation of a plethora of cell activities as transcription, gene silencing and chromosome opening, for example. Thus, herein we present an in silico approach for analyze the modulation of lncRNAs by R. amblyommii in A. sculptum tick. Using previously published data sets of A. sculptum transcriptomes, we identified putative lncRNAs and evaluated for differential expression in midgut and ovaries in this tick specie. Two assemblers were tested, Trinity and CLC Genomics, to construct the contigs and a pipeline to select the sequences with lncRNA characteristics. We obtained 31,888 and 23,733 contigs of putative lncRNAs using Trinity and CLC Genomics, respectively. We further identified more than 500 sequences of putative lncRNA that significantly aligned to sequences of RefSeq RNA, RNA Central and NONCODE databases. The transcriptome analysis further suggests that R. amblyommii is inducing differential expression of putative lncRNAs in midgut and ovary tissues. The work herein contributes for tick lncRNA database increasing and the initials insights of which lncRNA sequences are involving in Rickettsia-tick relationship

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2016-03-21T14:59:22Z No. of bitstreams: 1 texto completo.pdf: 3716003 bytes, checksum: 3011b5cf3648f69aa355d3ce6312b5c6 (MD5) / Made available in DSpace on 2016-03-21T14:59:22Z (GMT). No. of bitstreams: 1 texto completo.pdf: 3716003 bytes, checksum: 3011b5cf3648f69aa355d3ce6312b5c6 (MD5) Previous issue date: 2015-06-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / As lipocalinas constituem uma família de pequenas proteínas extracelulares com cerca de 200 resíduos de aminoácidos e massa molecular de 20 Kda possuem forte divergência em suas sequências de aminoácidos mas mantêm a sua estrutura tridimensional estritamente conservada. Sua diversidade funcional e reconhecimento molecular, contribuem para a utilização dessas proteínas como base para o desenvolvimento de alguns biofármacos. Os genes constitutivos participam de diversas funções do metabolismo celular e são utilizados como controle endógeno sendo fundamentais para validação dos resultados no estudo da expressão utilizando real time RTq-PCR. Em relação a esses genes, ainda não foram publicados trabalhos que identifiquem e validem genes constitutivos como genes de referência para a espécie Amblyomma sculptum (Complexo Amblyomma cajennense). Atualmente, transcriptomas de órgãos e tecidos de carrapatos do gênero Amblyomma têm sido publicados identificando uma variedade de transcritos de lipocalinas com domínio de ligação a histamina e serotonina (SHBP). Neste trabalho, realizamos por predição in silico a identificação de nove lipocalinas com domínio de ligação a serotonina e histamina estabelecendo possíveis relações filogenéticas funcionais de transcritos de lipocalinas identificadas no transcriptoma de glândula salivar do A. sculptum. Realizamos o desenho e a validação de 3 primers de genes de lipocalinas identificadas, lip4, lip7 e lip24. Foram analisados e validados 3 primers de genes constitutivos (g6pdh, 3ohcoadh e gapdh) candidatos potenciais a genes de referência em glândulas salivares, intestinos e ovários do carrapato A. sculptum em situação de não infecção/infecção com R. amblyommii. Foram validados 3 modelos estruturais correspondentes às proteínas lipocalina 4, lipocalina 7 e lipocalina 24. E ainda, realizou-se a análise da expressão diferencial de lipocalinas e potenciais genes de referência em glândula salivar, intestino e ovário de A. sculptum. Os dados obtidos na análise de expressão gênica por real time RT-qPCR suportam a hipótese de que os três genes de lipocalinas interferem na interação microrganismo-vetor sendo necessários trabalhos adicionais para verificação da validade desta hipótese, uma vez que dentre as 121 lipocalinas preditas no transcriptoma de glândulas salivares, somente nove apresentam domínios SHBP sugerindo um papel especial na função destas nos tecidos estudados. / The lipocalin’s family compound a small extracellular protein of about 200 amino acid residues and a molecular mass of 20 kDa present strong divergence in their amino acid sequences but retain its strictly conserved three dimensional structure. Its functional diversity and molecular recognition, contributing to the use of these proteins as a basis for the development of some biopharmaceuticals. The reference genes involved in various cellular metabolism functions and are used, as endogenous control is necessary to validate the results in the study of the expression for real RTQ-PCR analisys. For these genes have not yet been published works to identify and validate constitutive genes as the reference genes for Amblyomma sculptum (Amblyomma cajennense complex) species. Currently, transcriptomes of organs and tissues of the genus Amblyomma ticks have been published identifying a variety of lipocalins transcripts with binding domain histamine and serotonine (SHBP). We carried out by prediction in silico identification of nine lipocalins with serotonine and histamine binding domain setting possible functional phylogenetic relationships of lipocalins transcripts identified in salivary gland transcriptome of A. sculptum. We carry out the design and validation of three primers identified lipocalin genes, lip4, lip7 and lip24. They were analyzed and validated three primers of constitutive genes (g6pdh, 3ohcoadh and gapdh) potential candidate reference genes in the salivary glands, ovaries and intestines A. sculptum tick for a situation of no infection / infection with R. amblyommii. Were validated three corresponding structural models to lipocalins 4, 7 and 24. Yet, there was the analysis of differential expression of lipocalins and potential reference genes in the salivary gland, gut and ovarian A. sculptum tick. The results of analysis in experiment of gene expression by real time RT-qPCR support the hypothesis that the three lipocalins genes interfere with the microorganism-vector interactions. Finaly it is necessary a further work to verify the validity of this hypothesis, since among the 121 lipocalins predicted in transcriptome salivary glands, only nine have SHBP domains suggesting a special role in the function of these tissues studied.

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-30Bitstream added on 2014-06-13T20:05:16Z : No. of bitstreams: 1 veronez_va_dr_jabo.pdf: 622519 bytes, checksum: 8c09bda0244e349ed6030a5dfb6aedd7 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Cerrado, considerado a savana sul-americana, abrange cerca de dois milhões de km2 e é muito rica em espécies endémicas, mas ameaçada pela agricultura. Neste trabalho são apresentadas espécies de carrapatos e sua distribuição sazonal e relativa dentro das diferentes fitofisionomias na Estação Ecológica do Panga, uma pequena reserva de Cerrado, no Estado de Minas Gerais, Brasil. Ao longo de dois anos foram coletados do ambiente 2.694 carrapatos. Destes, 73,5% eram da espécie Amblyomma cajennense e 0,6%. Amblyomma dubitatum Todos os outros carrapatos (25,9%) eram formas imaturas e caracterizados como Amblyomma spp. Os carrapatos adultos de A. cajennense apresentaram picos numéricos na primavera e as ninfas no inverno dos dois anos. Bolos de larval Amblyomma spp foram encontrados no outono e inverno. Os carrapatos adultos (46,7%) e ninfas (39,5%) foram mais freqüentemente encontrados em matas, enquanto a maioria dos grupos de larvas foram encontradas nas veredas ou próximo a elas (39%). Amblyomma cajennense, Anocentor nitens, Rhipicephalus (Boophilus) microplus and Rhipicephalus sanguineus foram encontrados em animais domésticos de propriedades vizinhas. Não foram encontradas formas características de Rickettsia na hemolinfa de 497 A. cajennense e de um A. dubitatum. Os resultados confirmam relatos anteriores sobre a prevalência esmagadora de carrapatos A. cajennense no bioma Cerrado do Brasil e acrescentou informações sobre as preferências de habitat desta espécie de carrapato, um vetor importante no Brasil da febre maculosa. / Cerrado biome, the South American savannah, covers about two million km2 and is very rich in endemic species but threatened by agriculture. In this work free-living tick species are presented, and their seasonal and relative distribution within the various phytophysiognomies in a Stacey Ecologic Panga, small Cerrado reserve, in Minas Gerais State, Brazil. Overall 2,694 free-living ticks were found during a two years sampling period with CO2 traps and cloth dragging. Of these, 73.5% were Amblyomma cajennense and 0.6% Amblyomma dubitatum. All other ticks (25.9%) were retained as Amblyomma spp. Adults of A. cajennense peaked in spring, the nymphs in winter of both years. Amblyomma larval clusters were found in autumn and winter. Adult ticks (46.7%) and nymphs (39.5%) were most often found in woodlands, whereas most larval clusters were found in valley-side marshes (39%). Amblyomma cajennense, Anocentor nitens, Rhipicephalus (Boophilus) microplus and Rhipicephalus sanguineus ticks were found on domestic animals from neighboring properties. Search for Rickettsia in the hemolymph of 497 A. cajennense and one A. dubitatum ticks yielded negative results. Results confirmed earlier reports on the overwhelming prevalence of A. cajennense ticks in the Cerrado biome of Brazil and added information to habitat preferences of this tick species, a major vector in Brazil of the Rocky Mountain spotted fever.

Rabbits and goats were inoculated with crude, membrane-associated and soluble components extracted from unengorged adult females and nymphs of the bont tick Amblyomma hebraeum. Inoculation provided some protection against nymphal infestation, however it had little effect on adult feeding. Histological examination of adults fed on inoculated hosts showed evidence of gut damage. Skin provocation testing with tick extracts elicited a Type I immediate hypersensitivity which was influenced by antihistamine. A delayed skin reaction was also evident. Whether this was attributable to Type III Arthus reaction or Type IV cell-mediated hypersensitivity was not determined. A comparative histological study of sites of tick extract injection, on inoculated and naive hosts, demonstrated the role of eosinophils in the hosts response to tick feeding. Serological examination revealed elevated anti-A hebraeum lgG titres following inoculation. These titres were found to decrease in the ten weeks after inoculation, despite the hosts being repeatedly infested with A hebraeum. Although the IgG titres of naive control hosts increased after each tick infestation, they failed to reach the titres achieved through inoculation. Western blot analysis of serum from inoculated hosts recognized most of the A. hebraeum proteins against which it was screened.

No description available.