Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-09-20T19:27:14Z No. of bitstreams: 1 fernandacalmon.pdf: 376302 bytes, checksum: f059fa7ccf2af6362e23df2ba62301fb (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-09-22T15:20:06Z (GMT) No. of bitstreams: 1 fernandacalmon.pdf: 376302 bytes, checksum: f059fa7ccf2af6362e23df2ba62301fb (MD5) / Made available in DSpace on 2017-09-22T15:20:06Z (GMT). No. of bitstreams: 1 fernandacalmon.pdf: 376302 bytes, checksum: f059fa7ccf2af6362e23df2ba62301fb (MD5) Previous issue date: 2013-02-07 / O presente estudo objetivou avaliar a toxicidade de diferentes solventes e do Tween 80 sobre os estágios imaturos de Rhipicephalus sanguineus e sobre larvas e ninfas ingurgitadas e ninfas não ingurgitadas de Amblyomma cajennense. Para larvas não ingurgitadas de R. sanguineus foi utilizado o teste de pacote de larvas modificado e a mortalidade foi avaliada após 24 horas. O mesmo teste foi adaptado para a realização do experimento com ninfas não ingurgitadas das duas espécies e nesses dois experimentos foram testados os solventes etanol, metanol e acetona puros e dimetilsufóxido e tween 80 na concentração de 50μl/ml. Em relação aos testes com larvas e ninfas ingurgitadas foi utilizada a metodologia de imersão e a mortalidade foi avaliada após 15 dias. Os solventes etanol, metanol e acetona foram testados nas concentrações de 50, 250, 500, 750 e 1000 μl/ml, enquanto que o solvente dimetilsufóxido e o surfactante Tween foram testados somente a 50μl/ml, no entanto, para A.cajennense o surfactante foi acrescido com a concentração de 25μl/ml para os estágios ingurgitados. Para cada teste foi feito um controle com água destilada. Foram feitas dez repetições por tratamento e os grupos experimentais foram mantidos em câmara climatizada (27±1°C e UR>80±10%). Para R. sanguineus a acetona se mostrou tóxica nos estágios ingurgitados na maior concentração, o etanol e metanol foram tóxicos na maior concentração apenas para larvas ingurgitadas; para as larvas ingurgitadas de A. cajennense o etanol, metanol e a acetona forma tóxicos na maior concentração e o Tween matou 52 e 80% das larvas para as concentrações de 25 e 50μl/ml respectivamente, já para ninfas ingurgitadas o metanol matou 17,4 e a acetona 100% quando testados em qualidade pura, as ninfas não ingurgitadas não demonstraram sensibilidade aos solventes testados. Quanto a observação preliminar sobre a presença de ectoparasitos em aves silvestres, foram examinadas 58 aves de 5 famílias com prevalência de 15,5% de ectoparasitos. / The present study aimed to evaluate the toxicity of different solvents and of Tween 80 in immature stages of Rhipicephalus sanguineus and in larvae and engorged and non engorged nymphs of Amblyomma cajennense. For non engorged larvae of R. sanguineus it was utilized the larval packet test modified and mortality was evaluated after 24 hours. The same test was adapted for non engorged nymphs of both species and in both assays were tested ethanol, methanol and acetone in its pure concentrations and dimethylsulphoxide and Tween 80 in a concentration of 50μl/ml. For engorged larvae and nymphs it was utilized the immersion test and mortality was available after 15 days. The solvents ethanol, methanol and acetone were testes at concentrations of 50, 250, 500, 750 and 1000 μl/ml, while dimethylsulphoxide and the surfactant Tween were tested at concentrations of 50μl/ml, however, in the assays with A.cajennense engorged stages, the surfactant was added with a concentration of 25μl/ml. Ten repetitions were made for each treatment and the experimental groups were maintained in a BOD incubator (27±1°C e UR>80±10%).For each test it was made a control group with distilled water. For R.. sanguineus acetone showed toxic effects in engorged stages at the higher concentration, ethanol and methanol were toxic at higher concentration only for engorged larvae; to engorged larvae of A. cajennense the ethanol, methanol and acetone were toxic in the higher concentrations and the Tween caused mortality of 52 and 80% of larvae in concentrations of 25 and 50μl/ml respectively, for engorged nymphs, methanol caused a mortality of 17,4 and acetone of 100% when tested in pure concentrations, the tested solvents did not show any effect in engorged nymphs. As a preliminary remark on the presence of ectoparasites in wild birds, 58 birds were examined from 5 families with a prevalence of 15.5% of ectoparasites.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2015-12-02T19:00:12Z No. of bitstreams: 1 laryssaxavieraraujo.pdf: 1452466 bytes, checksum: 171c39ef6b34539f22d4565011471c2b (MD5) / Rejected by Adriana Oliveira (adriana.oliveira@ufjf.edu.br), reason: Título: somente primeira letra da primeira palavra em maiúsculo e nomes próprios também. Palavras-chave: primeira letra de cada palavra em maiúsculo. Acrescentar uma em cada linha (não precisa colocar ponto e nem vírgula no final) on 2015-12-03T11:58:48Z (GMT) / Submitted by Renata Lopes (renatasil82@gmail.com) on 2015-12-03T13:09:46Z No. of bitstreams: 1 laryssaxavieraraujo.pdf: 1452466 bytes, checksum: 171c39ef6b34539f22d4565011471c2b (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2015-12-03T13:58:23Z (GMT) No. of bitstreams: 1 laryssaxavieraraujo.pdf: 1452466 bytes, checksum: 171c39ef6b34539f22d4565011471c2b (MD5) / Made available in DSpace on 2015-12-03T13:58:23Z (GMT). No. of bitstreams: 1 laryssaxavieraraujo.pdf: 1452466 bytes, checksum: 171c39ef6b34539f22d4565011471c2b (MD5) Previous issue date: 2015-02-12 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Substâncias de origem vegetal como óleos essenciais e seus componentes têm demonstrado potencial para serem empregados no controle de diferentes pragas, incluindo carrapatos. Entre essas substâncias destacam-se os monoterpenos timol e carvacrol e o fenilpropanóide eugenol, moléculas que já tiveram sua atividade evidenciada sobre os carrapatos Rhipicephalus microplus e Rhipicephalus sanguineus s.l., espécies de grande importância econômica. Apesar da ação isolada dessas substâncias sobre carrapatos ter sido demonstrada in vitro em diferentes trabalhos, até o momento não existem estudos sobre o efeito da combinação das mesmas e relato de possível ação sinérgica sobre ixodídeos. Também são escassos os trabalhos que avaliam a atividade dessas substâncias em ambientes naturais ou semi-naturais, visando controlar carrapatos com ações direcionadas sobre a fase não parasitária. Dessa forma, este estudo teve como objetivos, avaliar os efeitos das combinações dos monoterpenos timol e carvacrol e do fenilpropanóide eugenol sobre larvas de R. microplus e R. sanguineus s.l. e avaliar a atividade carrapaticida do timol sobre larvas de R. microplus em condições semi-naturais. O primeiro capítulo traz os resultados dos efeitos das combinações das substâncias sobre as larvas desses carrapatos e para esse fim, foi utilizado o teste de pacote de larvas. Primeiramente foi feito o cálculo de CL50 e em seguida essas substâncias foram testadas em associação ou isoladas em concentrações subletais. Para larvas de R. microplus foram observados valores de CL50 para o timol, carvacrol e eugenol de 1,53; 1,76 e 4,67 mg/mL, respectivamente, e sinergismo entre todas as nove combinações testadas. Os valores de CL50 do timol, carvacrol e eugenol observados para larvas de R. sanguineus s.l., foram de 2,98; 3,29 e 5,19 mg/mL, respectivamente, sendo esse o primeiro estudo a determinar a CL50 dessas substâncias para R. sanguineus s.l. Para esta espécie, oito misturas apresentaram efeito sinérgico e apenas a combinação de carvacrol+timol na concentração da CL50, apresentou efeito sinérgico moderado. No segundo capítulo foi observado, pela primeira vez na literatura, que o timol ao ser aplicado em mudas de Brachiaria decumbens infestadas artificialmente com larvas de R. microplus foi capaz de reduzir 11 significativamente a infestação por esse carrapato. As maiores concentrações (10, 15, 20, 25 e 30,0 mg/mL) ocasionaram eficácias acima de 95% e os valores encontrados para as CL50 e CL90 foram de 3,45 e 9,25 mg/mL, respectivamente. Conclui-se que associações entre timol, carvacrol e eugenol em concentrações subletais apresentam efeito sinérgico sobre larvas de R. microplus e R. sanguineus s.l. e que o timol apresenta potencial para redução de larvas de R. microplus em condições semi-naturais. / Substances of plant origin, such as essential oils and their components have demonstrated the potential to be used to control different pests, including ticks. Among these substances stand out monoterpenes thymol and carvacrol and the phenylpropanoid eugenol, molecules that already had their activity demonstrated against ticks Rhipicephalus microplus and Rhipicephalus sanguineus s.l., species of great economic importance. Despite the isolated action of these substances on ticks has been demonstrated in vitro in different papers, until now there are no studies on the effect of the combination of this substances and report of possible synergistic action on ticks. Also there are few studies that evaluate the activity of these substances in natural and semi-natural environments in order to control ticks with targeted actions on the non-parasitic phase. Therefore, this study aimed to evaluate the effects of combinations of monoterpenes thymol and carvacrol and phenylpropanoid eugenol on larvae of Rhipicephalus microplus and Rhipicephalus sanguineus s.l. and evaluate the insecticidal activity of thymol on larvae of R. microplus in semi-natural conditions. The first chapter presents the results of the effects of combinations of substances on the larvae of these ticks. For R. microplus larvae were observed LC50 values for the thymol, carvacrol and eugenol of 1.53; 1.76 and 4.67 mg/ml, respectively, and synergism between all combinations tested. The LC50 values of thymol, carvacrol and eugenol observed for larvae of R. sanguineus s.l., were 2.98; 3.29 and 5.19 mg/ml, respectively. This is the first work reporting the LC50 values of these three substances against R. sanguineus s.l. For this species, eight combinations showed synergistic effect and only the combination of carvacrol+thymol in the concentration of LC50, showed moderate synergism. In this study it was observed that the larvae of R. microplus were more sensitive to the tested substances and their combinations. In the second chapter it was observed for the first time that thymol when applied in Brachiaria decumbens seedlings artificially infested with larvae of R. microplus was able to significantly reduce the infestation by this tick. The highest concentrations (10, 15, 20, 25 and 30.0 mg/ml) resulted efficiencies above 95% and the values found for the LC50 and LC90 were 3.45 and 9.25 mg/ml, respectively. It is concluded that associations between thymol, eugenol and carvacrol at sublethal concentrations have a synergistic effect on larvae of R. microplus and Rhipicephalus sanguineus s.l. and thymol has the potential for reducing R. microplus larvae in semi-natural conditions.

Tick-borne encephalitis is a vector-borne zoonotic disease with more than 12,000 annual clinical reported cases globally (WHO, 2021). Tick-borne encephalitis is caused by the flavivirus and transferred by Ixodes ricinus from roe deer to human and affects the central Nervous system. Climate change also increases tick-borne encephalitis incidences in Sweden and fluctuated considerably from year to year. A quantitative study design with secondary data was conducted to analyze the spatial and temporal pattern of Tick-borne encephalitis in Sweden from 1986-2020. The distribution of Tick-borne encephalitis within age and sex, along with other factors were also analyzed. The results showed that Tick-borne encephalitis with passing each year spread across the country. Male and age group, 50-59, have more incidence of Tick-borne encephalitis. Incidents were associated with climate conditions such as temperature and precipitation, which provided a favorable environment for Ixodes ricinus for its lifecycle activities, host searching, and disease transmission. Roe deer population, other vertebrates abundance, vaccination, population interest, economy, and land change are the critical factors that facilitate the disease incidence or control. People who visit forests for hunting, trekking, leisure, and professional activities without proper immunity and preventive measures are at risk to infect with Tick-borne encephalitis. The theory One health approach showed suitable performance for the control of this vulnerable climate zoonotic disease.

In Western Europe the incidence of tick-borne encephalitis (TBE) has increased over the last 30 years, coupled with changes in distribution. Modifications in the TBE enzootic cycle, through a combination of changes in temperature, vertebrate abundance and habitat suitability may have increased the risk of TBE in recent years. In Switzerland, analysis using satellite-derived climate data demonstrated that the environment of areas with TBE since the 1980s and areas that recently became endemic for TBE have become more similar between 2001 and 2009. This was coupled with an increase in April, May and June temperature, which could have affected the tick population and/or human exposure to ticks. Deer and boar abundance also changed in some cantons. In Germany, spatio-temporal modelling demonstrated the importance of temperature, vertebrate abundance and unemployment in the incidence and distribution of TBE between 2001 and 2009. Changes in TBE reporting, April, May and June temperature, vertebrate abundance and pesticide use may have contributed to increases in TBE in 1992 and 2001. Human exposure patterns, however, appear to be as important as the enzootic cycle in shaping the incidence of TBE, not only in determining the overall trend but also in interacting with the weekly, seasonal and yearly patterns of tick hazard to give the observed incidence. In Switzerland, in weeks with warm, sunny weather, human exposure to ticks is promoted and short-term increases in tick bites are seen. Human outdoor activity also shifts the seasonal pattern of tick bites, when compared with tick questing. There was no apparent increase in time spent in outdoor activities between the 1990s and 2000s in Italy, Germany and Austria, but survey data demonstrated that walking and hiking were already popular activities across Europe by the 1990s. The popularity of mushroom and berry foraging as a source of income in Latvia, Lithuania and Poland, coupled with the expense of vaccination, provide an inverse link between economic wellbeing and TBE risk. Correspondingly, in 2009, the economic recession was associated with an increase with TBE in these three countries.

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...

L’augmentation depuis une trentaine d’années des maladies infectieuses dites émergentes ou ré-émergentes chez l’homme, causées à plus de 70% par des pathogènes issus d’espèces hôtes animales (i.e. Ebola, SIDA), stimule l’étude de systèmes éco-épidémiologiques à l’interface entres populations humaines et animales (i.e. sauvages et/ou domestiques).Le contact entre hôtes est un phénomène important dans l’étude de ces systèmes car il permet la transmission des pathogènes entre individus et la diffusion de maladie au sein et entre populations. Nous avons choisi la maladie de la fièvre aphteuse comme modèle d’étude de la transmission de pathogènes des populations sauvages vers les populations domestiques. Le buffle africain (Syncerus caffer) étant le réservoir présumé de cette maladie fortement contagieuse, nous nous sommes interrogés sur les conditions de transfert au bétail (Bos taurus et Bos indicus) du virus aphteux aux frontières de trois parcs nationaux africains qui constituent des interfaces entre espaces anthropiques et protégés perméables aux mouvements d’animaux. Dans le cadre de ce doctorat 4 protocoles ont été mis en place entre 2010 et 2011 au Zimbabwe. Premièrement, des colliers GPS (Global Positionning System) ont été déployés sur des bovins sauvages/domestiques pour décrire leurs déplacements dans le paysage et quantifier les contacts interspécifiques. Des colliers furent également posés sur l’une des espèces prédatrices de ces ongulés: le lion (Panthera leo). L’intégration de la guilde des prédateurs nous a permis d’estimer les modifications de l’utilisation de l’espace par les herbivores en réponse à la présence de carnivores et les conséquences en termes de contacts et de transmission interspécifique de pathogènes. Deuxièmement, un suivi longitudinal sérologique sur le bétail a complété le protocole télémétrique avec des prélèvements répétés sur des individus marqués selon le cycle saisonnier. Troisièmement, les contacts au sein des populations de bovins domestiques ont été caractérisés par des enquêtes auprès des éleveurs. Quatrièmement, le rôle potentiel de la diversité des hôtes sur le risque infectieux d’un écosystème a été exploré par l’estimation de densité de macro-parasites dans le paysage selon une variation de la gamme d’hôtes potentiels (i.e. (i) sauvages, (ii) sauvages et domestiques et (iii) uniquement domestiques).Nos résultats montrent que (1) les taux d’interaction interspécifiques, estimés par télémétrie, varient entre sites et présentent une saisonnalité prononcée (i.e. pic saison sèche chaude). (2) La distribution des ressources conditionne la périodicité et la distribution de ces contacts dans les différents compartiments du paysage. (3) La fréquence des incursions du bétail dans un espace protégé ainsi que les taux de contacts avec les buffles influencent positivement la probabilité d’acquisition d’anticorps anti-aphteux chez le bétail. La probabilité de perte d’anticorps est également fonction du niveau d’interaction avec les buffles mais selon une relation négative. (4) La densité du réseau d’interaction intra-spécifique domestique influence positivement l’incidence sérologique de la fièvre aphteuse. (5) La présence de prédateurs supérieurs dans le paysage permettrait de limiter les incursions du bétail dans les espaces protégés et diminuerait la probabilité d’infection par les populations d’hôtes sauvages. (6) Enfin les densités de macro-parasites dans la végétation sont supérieures dans des espaces communaux sans interaction avec les populations sauvages et où la richesse spécifique des hôtes est plus faible. Les résultats de cette étude sur la transmission interspécifique de pathogènes entre populations sauvages et domestiques dans les écosystèmes tropicaux ouvrent des champs de réflexion encore largement inexplorés, notamment sur l’évolution de la virulence et des modes de transmission des pathogènes ayant comme hôtes des populations sympatriques sauvages et domestiques. / Emerging or re-emerging diseases in human populations have increased over the last thirty years. Since 70% of such diseases are caused by pathogens originating from animal hosts (i.e. Ebola, AIDS, and avian influenza), this increase has prompted the study of eco-epidemiological systems that occur at the interface between human and animal populations (i.e. wild and/or domestic). Contacts between hosts are particularly important factors in these systems since they result in pathogen transmission among individuals and, therefore, disease diffusion within and among populations. We used foot-and-mouth disease (FMD) as a model to study pathogen transmission from wild to domestic populations. As the African buffalo (Syncerus caffer) is the presumed reservoir of this highly contagious disease, we examined the conditions in which the virus was transmitted to cattle sensitive to the disease (Bos taurus and Bos indicus) at the borders of African national parks; these areas are interfaces between anthropogenic and protected areas in which animals can move freely.In the context, 4 protocols were implemented between 2010 and 2011 in Zimbabwe. First, GPS (Global Positioning System) collars were placed on cattle and buffalo in order to describe and analyze their movements across the landscape as well as to quantify interspecific contacts. In one of the study sites, collars were attached to one of the predators of these ungulates: the lion (Panthera leo). By integrating the predator guild into our telemetry protocols, we could examine the potential changes in spatial use by cattle and buffalo in response to predator presence and their consequences for contact dynamics and interspecific pathogen transmission. Second, a longitudinal serological survey was conducted in which tagged individuals were sampled repeatedly over the course of different seasons. Third, to characterize contacts within the domestic host population, interviews were conducted with cattle owners regarding their husbandry practices across seasons. Fourth, to describe the potential role of host diversity in ecosystem infection risks, macroparasite density (i.e. ticks) was estimated for landscape compartments that contrasted in terms of potential hosts present (i.e. (i) wild, (ii) domestic and wild, and (iii) domestic only).Our study primarily shows the following results. (1) Interspecific interaction rates, as estimated by telemetry, vary between sites and have a pronounced seasonality (peak occurs during the hot dry season). (2) Resource distribution (i.e. water and grazing areas) seems to condition the frequency and distribution of these contacts in the different landscape compartments. (3) Cattle incursion frequencies into protected areas and the frequency and intensity of contact with buffalo significantly positively affect the probability of foot-and-mouth antibody acquisition in cattle. The probability of antibody loss in cattle is also a function of diminished rates of interaction with buffalo. (4) Intraspecific interaction densities positively influence FMD serological incidence in cattle. (5) Top predator presence in the landscape could limit cattle incursion into protected areas and reduce the likelihood of their being infected by wild host populations. (6) Finally, the estimated densities of macroparasites in the vegetation were higher in communal spaces where there was no interaction with wild hosts and where host species richness was weak.The results of this study on the interspecific transmission of pathogens between wild and domestic populations in tropical ecosystems encourage the exploration of research topics that are still largely unexplored, including the evolution of virulence transmission modes of pathogens hosted by sympatric wild and domestic populations.

Infestações com Rhipicephalus microplus, o carrapato dos bovinos, causam enormes prejuízos econômicos para a pecuária. Os carrapatos estão desenvolvendo resistência aos carrapaticidas que, além dessa desvantagem, deixam resíduos em carne e leite. Vacinas anticarrapato representam uma alternativa sustentável de controle de infestações, mas as atualmente disponíveis têm efeitos parciais e transitórios. Surge, assim, a necessidade de identificar novos antígenos vacinais. Para alcançar esse objetivo este trabalho explora o fato de que bovinos apresentam fenótipos contrastantes e herdáveis de infestações que são específicos de certas raças. Além disso, o nível de imunidade do hospedeiro afeta a transcrição de genes de glândulas salivares do carrapato, órgão que produz proteínas que medeiam o parasitismo. A hipótese de trabalho é a que os diferentes níveis da imunidade anticarrapato do hospedeiro afetam, também, a composição salivar do parasita. Assim, em carrapatos alimentando-se em hospedeiros resistentes as proteínas que são cruciais ao parasitismo poderão estar ausentes ou deficientes na sua saliva e por isso os carrapatos não terminam sua refeição de sangue. A neutralização dessas mesmas proteínas pela imunidade humoral pode ter o mesmo efeito e por isso, essas proteínas constituem bons antígenos vacinais. Assim, o objetivo do trabalho foi identificar novos antígenos vacinais em saliva de fêmeas e glândulas salivares de ninfas, machos e fêmeas de carrapatos alimentados em hospedeiros resistentes e suscetíveis, bem como em larvas não alimentadas oriundas de ovos de fêmeas alimentadas nestes mesmos hospedeiros. Para isso, foram empregadas abordagens de sequenciamento de nova geração \"RNA-Seq\" (454) e abordagens proteômicas, como análise diferencial em gel (DIGE) e Western Blots (imunoproteoma) seguido de sequenciamento de massa, além da tecnologia de identificação de proteínas multidimensionais (ou Multidimensional Protein Identification Technology, MudPIT) para descrever o proteoma das glândulas salivares e da saliva de fêmeas. A análise transcriptômica resultou no sequencimanto de 1.999.086 reads que permitiu identificar e classificar 11.676 sequências codificadoras (CDS), muitas das quais (3.600 CDS) contêm peptídeo sinal que é indicativo de secreção, portanto podendo estar presente na saliva e Resumo Gustavo Rocha Garcia apresentar função importante na hematofagia. Por meio de MudPIT, identificamos 321 proteínas salivares diferentes, além de 126 proteínas no DIGE e 266 proteínas nos imunoproteomas. Muitas dessas proteínas podem ser consideradas antígenos potenciais por estarem associadas com a hematofagia/parasitismo, tais como proteases, nucleases, inibidores de proteases, peptídeos antimicrobianos, proteínas de fixação, entre outros, inclusive proteínas ainda não caracterizadas. A maioria dos genes codificantes dessas proteínas está mais expressa em carrapatos alimentados em hospedeiros suscetíveis, principalmente em carrapatos machos. Além disso, muitas dessas proteínas não são reconhecidas por soros bovinos, inclusive soros de bovinos infestados, embora soros de bovinos infestados e resistentes ao carrapato apresente a maioria das reatividades. O conjunto dos resultados sugere que em nível de proteína a composição da saliva também é afetada pelos diferentes níveis de imunidade dos hospedeiros, além de variar com o ciclo de vida do carrapato. Desse modo, concluímos que as estratégias de investigação empregadas foram satisfatórias para identificar um conjunto de antígenos salivares do carrapato R. microplus que representam proteínas alvos para compor vacinas multicomponentes anticarrapato. / Infestation with Rhipicephalus microplus, the cattle tick, causes huge economic losses to livestock. Ticks are developing resistance to acaricides that, besides this disadvantage, leave residues in meat and milk. The anti tick vaccines represent a sustainable alternative of the infestations control, but the currently available has partial and transient effects. Thus arises the need to identify new vaccine antigens. To achieve this goal, this work explores the fact that cattle exhibit contrasting phenotypes and inheritable of infestations that are specific to certain breeds. Furthermore, the level of immunity of the host affects gene transcription tick salivary gland, organ that produces proteins that mediate the parasitism. The working hypothesis is that different levels of anti tick immunity of host affect also the salivary composition of the parasite. So in ticks feeding on resistant hosts the proteins that are crucial to parasitism may be absent or deficient in their saliva, and by this the ticks do not finish your meal blood. The neutralization of these same proteins by humoral immunity can have the same effect and by this, these proteins are good vaccine antigens. So, the aim of the study was to identify new vaccine antigens in saliva from females and salivary glands of nymphs, males and females of ticks fed on resistant and susceptible hosts as well as in unfed larvae originating from eggs of females fed on these same hosts. To this, were employed sequencing approaches of new generation \"RNA-Seq\" (454) and proteomic approaches, such as differential analysis in gel (DIGE) and Western Blots (immunoproteomics) followed by sequencing mass, besides the Multidimensional Protein Identification Technology (MudPIT) to describe the proteome of the salivary glands and saliva of females. The transcriptomics analysis identified 11,676 coding sequences (CDS), many of which (3,600 CDS) contain predicted signal peptide indicative of secretion, therefore may be present in saliva and provide an important function in blood feeding. Through MudPIT, we identify 321 different salivary proteins, besides 126 proteins in DIGE and 266 proteins in immunoproteomics. Many of these proteins may be considered as potential antigens to be associated with the blood meal/ parasitism, such as proteases, nucleases, protease inhibitors, antimicrobial peptides, proteins of attachment, among Abstract Gustavo Rocha Garcia others, including proteins not yet characterized. Most of the genes encoding of these proteins are more expressed in ticks fed on susceptible hosts, especially in male ticks. Moreover, many of these proteins are not recognized by bovine sera, including sera from infested hosts, although sera from infested and resistant host to tick present the most reactivities. The overall results suggest that in protein level, the composition of saliva is also affected by the different levels of immunity of the host, besides vary with the tick life cycle. Thus, we conclude that the research strategies employed were satisfactory to identify a set of tick salivary antigens from R. microplus that represent target proteins for composing anti tick multicomponent vaccines.

This thesis is focused on the role of host cytoskeleton, primarily microtubules and microfilaments, during tick-borne encephalitis virus infection in human neuroblastoma cell line SK-N-SH and tick cell line IRE/CTVM19. The importance of cytoskeletal integrity and dynamics to the viral replication cycle were examined using specific chemical inhibitors showing the virus utilizes studied structures in both cell lines. Immunofluorescence microscopy revealed structural changes in the actin cytoskeleton during late infection in SK-N-SH cells. Moreover, differences in expression of cytoskeleton-associated genes in both cell lines were compared. Several genes with up-regulated expression in SK-N-SH cells were identified during late infection.

No description available.

Glycosylation in vertebrates has a main role in many important processes such as cell transport, protein folding, secretion of proteins etc. What function has glycosylation in arthropods, for example in ticks, is rarely studied. This work was focused on analysis of N-glycans in tick tissues, namely in Ixodes ricinus and Dermacentor marginatus. High-mannose glycans as well as complex glycans with or without core-fucosylation were identified in this study.Furthermore several sialylated glycans were present in the studied samples. Sialic acid is found in arthropods rarely and this is the first study which directly proves its presence in ticks using mass spectrometry.