Existe atualmente uma restrição muito forte ao uso de antibióticos como promotores de crescimento na alimentação avícola, uma vez que tais produtos passaram a ser vistos como fatores de risco para a saúde humana. Estudos com produtos alternativos tornaram-se necessários e, neste sentido, as leveduras vem sendo exploradas, principalmente, seu conteúdo celular. Dessa forma, o presente estudo teve como objetivo avaliar o uso de levedura hidrolisada, fonte de nucleotídeos, sobre o desempenho e imunidade de frangos de corte, suplementados no período de 1 a 14 dias de idade. Foram utilizados 576 pintos de corte, machos, linhagem Ross 308, criados até 42 dias de idade sobre cama reutilizada. O delineamento foi inteiramente casualizado com 6 tratamentos: controle negativo (sem suplementação); controle positivo (com antibiótico) e diferentes níveis de adição de levedura hidrolisada (0,25%; 0,50%; 0,75% e 1,00%), sendo 8 repetições/tratamento. Considerando-se o período total de criação, foram observados efeitos significativos para os parâmetros de ganho de peso (GP) e conversão alimentar (CA). Por sua vez, o parâmetro de consumo de ração (CR) não apresentou efeitos significativos entre os tratamentos. Com relação à imunidade, a suplementação com levedura hidrolisada apresentou efeito sobre a resposta a vacinação contra a doença de Newcastle, produção de água oxigenada, óxido nítrico e influenciou também no peso relativo dos órgãos linfóides, com destaque para o nível 0,75% de suplementação. Diante dos resultados obtidos no presente estudo, pode-se concluir que a levedura hidrolisada, suplementada durante o período de 1 a 14 dias de idade, promove efeitos significativos no desempenho durante todo o período de criação e também influencia na imunidade das aves. / There is currently a strong restriction on the usage of antibiotics as growth promoters in broiler feed, since such products were seen as risk factors for human health. Research with alternative products has became necessary, and the yeast has been exploited, particularly its cellular content. Thus, this experiment was aimed at evaluating the use of hydrolyzed yeast, source of nucleotides on performance and immunity of broiler chickens supplemented during the period of 1 to 14 days old. A total of 576 male chicks, Ross 308 were raised up to 42 days old, on reused litter. The design was completely randomized with six treatments: negative control (no supplementation), positive control (with antibiotic) and different levels of hydrolyzed yeast (0.25%; 0.50%; 0.75% e 1.00%), with 8 replicates per treatment. Considering the whole breeding period, significant effects were observed for parameters of weight gain (WG) and feed conversion (FC). However, the parameter of feed consumption (FC) had no significant effect among the treatments. Regarding to immunity, the hydrolyzed yeast supplementation had an effect on the response to vaccination against Newcastle disease, production of hydrogen peroxide, nitric oxide and also influenced the relative weight of lymphoid organs, especially the 0.75% level of supplementation. With the results obtained in this experiment it is possible to conclude that the hydrolyzed yeast supplementation, during the period of 1 to 14 days old, promotes a significant effect on performance throughout the whole rearing period and also influences the immunity of the broiler chicks.

Foram utilizadas 750 poedeiras comerciais de linhagens brancas com 23 semanas ao início do período experimental, distribuídas num delineamento em blocos completos aleatorizados sendo estes caracterizados pela oncatenação das linhagens com coluna de gaiolas; os tratamentos foram cinco densidades ou taxas de lotação na gaiola (321,43; 375; 450; 562,50 e 750 cm2/ave) com seis repetições, totalizando 150 parcelas experimentais. As dietas experimentais foram à base de milho e farelo de soja, formuladas para suprir as exigências nutricionais das linhagens em todas as fases. O desempenho produtivo e econômico foram avaliados através do peso dos ovos (g), percentagem de postura (%), massa de ovos (g/ave/dia), consumo de ração (g/ave/dia), conversão alimentar por dúzia (kg/dz) e por quilo de ovo (kg/kg). A qualidade de ovos foi avaliada através da gravidade específica (GE), unidades Haugh (UH), percentagens de gema, albúmen e casca, espessura da casca (EC), resistência à quebra (RQ), coloração da gema e índice gema (IG). O bem-estar das aves foi avaliado através de indicadores clínicos, imunológicos e fisiológicos. Para efeito da avaliação dos resultados, foram estabelecidos seis blocos com repetições internas. Os dados foram analisados com auxílio do SAS, sob modelo misto, considerando os efeitos da densidade, período e a interação entre estes, como fixos, além dos efeitos aleatórios de bloco e resíduo. Por se tratar de medidas repetidas longitudinalmente, buscou-se a melhor estrutura de covariância para cada variável. Quando adequado foi aplicado o teste de tukey-kramer para a comparação de médias (p<0,05). O menor consumo de ração, CA/dz de ovos e melhor desempenho econômico, foram obtidos pela densidade 321,4 cm2/ave. Altas densidades de alojamento não influenciaram a qualidade interna de ovos, a % casca e RQ, mas essas variáveis tiveram efeitos dos períodos, com os melhores valores nos primeiros períodos. Altas densidades tiveram efeitos negativos sobre a GE e EC, principalmente no pico de postura das aves. Os ovos avaliados encontravam-se dentro dos valores desejados, mesmo nas maiores densidades, podendo ser classificados como de excelente qualidade. A densidade de alojamento não interferiru (p>0,05) na avaliação clínica das aves, mas o maior espaço proporcionado nas gaiolas do sistema covencional conferiu menor freqüência de lesões nas aves, indicando promoção e melhora ao seu bem-estar geral. Dados de freqüência cardíaca e temperatura da cloaca estavam dentro dos padrões de normalidade para a espécie. Não foram observadas diferenças (p>0,05) das densidades na resposta imunológica das aves, com os títulos dos anticorpos vacinais contra as doenças de Gumboro e Newcastle acima dos níveis do ponto de corte (cutoff). Os tratamentos não modificaram o perfil sanguíneo das aves e não foi possível caracterizar um padrão de corticosterona no plasma e de excreção de metabólitos fecais de glicocorticoides. A técnica de enzimoimunoensaio, empregada utilizando anticorpo primário contra corticosterona e anticorpo secundário anticoelho de cabra, foi capaz de detectar as variações nas concentrações de corticosterona nesses tecidos, contudo, o entendimento do significado desses achados ainda necessita de novas investigações. / 750 commercial laying hens of white strains aged 23 weeks at the beginning of the experimental period were used, distributed in a randomized complete block design, these being characterized by the concatenation of the strains with column of cages; the treatments were five densities or stocking rates in the cage (321,43, 375, 450, 562.50 and 750 cm2/bird) with six replications, amounting to 150 experimental plots. The experimental diets were on the basis of corn and soybean meal, formulated to meet the nutritional requirements of the strains in all the phases. Both the productive and economic performances were evaluated through the weight of the eggs (g), percentage of laying (%), egg mass (g/bird/day), feed intake (g/bird/day), feed conversion per dozen (kg/dozen) and per kg of egg (kg/kg). Egg quality was determined through specific gravity (SG), Haugh units (HU), percentages of yolk, albumen and shell, shell thickness (ST), resistance to breaking (BR), yolk coloration and yolk index (YI). The welfare of the birds was evaluated through clinical, immunological and physiological indicators. For the purpose of evaluation of the results, six blocks with internal replications were established. The data were analyzed with the aid of SAS, under a mixed model, considering the effects of density, period and the interaction between them, as fixed, in addition to the random effects of block and residue. For se tratar de measures longitudinally repeated, the best covariance structure for each variable was sought. When appropriate, the Tukey-Kramer test was used for comparison of means (p<0.05). The lowest feed intake, FI/dozen of eggs and best economic performance, were obtained at the density of 321.4 cm2/bird. High housing densities did not influence the internal quality of the eggs, % shell and BR, but these variables had effects from the periods, with the best values in the early periods. High densities had negative effects on SG and ST, mainly at the laying peak of the birds. The eggs checked lie within the desired values, even at the highest densities and can be classified as of excellent quality. The housing density did not interfere (p>0.05) in the clinical evaluation of the birds, but the greater space provided in the cages of the conventional system conferred a lower frequency of lesions in the birds, indicating promotion and improvement to their general welfare. Data of heart rate and cloacal temperature were within the normal limits for the species. No differences (p>0.05) of the densities in the immune response of the birds, with the titers of the vaccine antibodies against Gumboro and Newcastle diseases above the cutoff levels were found. The treatments did not modify the blood profile of the birds and it was not possible to characterize a standard of plasma corticosterone and excretion of fecal metabolites of glucocorticoids, but the enzyme immunoassay technique, using primary antibody against corticosterone and secondary goat anti-rabbit antibody, was capable of detecting variations in the corticosterone concentrations in these tissues, however, the understanding of the meaning of these findings still requires further investigation.

Atherosclerotic cardiovascular disease secondary to deposition of apolipoprotein B-containing lipoproteins in the artery wall is a leading cause of mortality. Therapies that reduce serum levels of atherogenic lipoprotein-cholesterol have been successful in reducing cardiovascular mortality, but this approach requires long-term treatment and substantial residual risk remains. Here, we investigate mechanistic determinants of atherosclerosis protection by two potential therapeutic approaches for lowering of residual cardiovascular risk. Using mouse models, we show that the nuclear receptor liver X receptor exerts an anti-inflammatory activity on innate immunity and atherosclerosis through both promotion of cholesterol efflux and a direct cis-repressive activity affecting neutrophil inflammation. We then assess the causal role of the cholesterol efflux pathway in human cardiovascular events by using genetic variants that modify high density lipoprotein-cholesterol in instrumental variable analysis. We show that this pathway is associated with protection from cardiovascular disease in a precise and robust Mendelian randomization analysis on an FDR-controlled set of variants, which suggests a causal effect. Thus, agents that target the cholesterol efflux and liver X receptor cis-repression pathways may be protective in atherosclerosis.

Le mélanome est la forme la plus agressive des cancers de la peau. Si sa prise en charge à des stades précoces est de bon pronostic, l’espérance de vie des patients chute dramatiquement pour les stades métastatiques. Malgré les avancées thérapeutiques spectaculaires récentes, le problème majeur réside dans la résistance aux traitements et la récidive et le défi principal est désormais de tendre vers un contrôle efficace et durable. Les anticorps monoclonaux (AcM) ont la capacité de cibler et éliminer spécifiquement la cellule tumorale tout en recrutant des cellules du système immunitaire, permettant de développer et/ou renforcer l’immunité de l’hôte avec le développement d’une réponse immune anti-tumorale de type vaccinale. Dans un modèle de tumeur solide de mélanome murin après greffe sous-cutanée des cellules B16F10, nous avons étudié le potentiel immunomodulateur de l’AcM TA99 qui cible un antigène de surface TYRP-1 surexprimé dans les mélanocytes tumoraux. Nos résultats montrent qu’environ 30% des souris sont protégées sur le long-terme et présentent une réponse immunitaire humorale et cellulaire mémoire. Par ailleurs, l’analyse de l’infiltrat immunitaire chez les souris qui échappent au traitement par l’AcM TA99 et qui développent une tumeur à plus ou moins long terme, montre une surexpression de PD-1 et Tim3 associée à une perte de fonctionnalité des cellules effectrices au sein de la tumeur. Ce même phénotype a été observé sur des biopsies de patients atteints de mélanome métastatique. Nous montrons aussi dans le cadre de ce travail que, le blocage de l’axe PD1/PD-L1 par inoculation d’un AcM anti-PD1 au moment de l’échappement, potentialise la réponse immunitaire anti-tumorale et entraîne une augmentation de la survie. Cependant, l’absence de régression complète suggère la mise en place d’autres voies immunosuppressives. En effet nous avons observé une surexpression des ectonucleotidases CD39 et CD73 dans le micro-environnement tumoral suggérant l’implication de l’adénosine dans la résistance au traitement de l’AcM TA99 plus l’α-PD-1. Ce résultat ouvre des perspectives intéressantes pour le blocage concomitant la voie de l’adénosine et de l’axe PD1/PD-L1. Une autre stratégie a consisté à améliorer les effets immunomodulateurs précoces de l’AcM TA99 en le combinant avec l’oxaliplatine, chimiothérapie favorisant la mort immunogénique. Bien que les combinaisons thérapeutiques testées dans cette étude montrent des effets in vivo encourageants avec un délai significatif dans la survie globale, aucune augmentation significative de la réponse anti-tumorale sur le long terme n’a pu être observée, suggérant la mise en place d’autres voies immunosuppressives non redondantes ou des stratégies de combinaisons non adaptées. Une analyse dynamique approfondie, tant phénotypique que fonctionnelle, des différents acteurs cellulaires du micro-environnement tumoral sera une étape clé dans la mise en place de combinaisons pertinentes en association avec l’AcM TA99. Ce travail prend d’autant plus d’intérêt qu’un essai clinique de phase I (IMC-20D7S) utilisant le flanvotumab (équivalent humain de l’AcM TA99) réalisé chez 27 patients atteints de mélanome métastatique, montre des effets cliniques intéressants sans effets secondaires sévères, ouvrant la voie au développement de combinaisons thérapeutiques associées à cet AcM. / Melanoma is the most aggressive form of skin cancer. Although early management is of good prognosis, the survival of patients decrease dramatically for metastatic stages. Despite the recent spectacular therapeutic advances, the major problem lies in resistance to treatment and relapse and the main challenge now is to develop an effective and sustainable control. Monoclonal antibodies (mAbs) have the ability to specifically target and eliminate tumor cells while recruiting cells from the immune system, to develop and / or enhance the immunity of the host with the development of a vaccinal immune response. In a solid tumor model of murine melanoma after subcutaneous transplantation of B16F10 cells, we investigated the immunomodulatory effect of TA99 mAb targeting a TYRP-1 surface antigen overexpressed in tumor melanocytes. Our results showed that about 30% of mice are protected in the long term and have an antitumoral humoral and cellular immune response. Moreover, the analysis of the immune infiltrate in mice that escape to the treatment with TA99 mAb and develop a tumor, shows an overexpression of PD-1 and Tim3 associated with a loss of effector cell functions within the tumor. This same phenotype has been observed in biopsies of patients with metastatic melanoma. Thus, blocking the PD-1 / PDL-1 axis by inoculation of an anti-PD1 mAb at the time of tumor escape potentiates the anti-tumor immune response and results in increased survival. However, the absence of complete regression suggests the establishment of other immunosuppressive pathways. Indeed we have observed an overexpression of CD39 and CD73 ectonucleotidases in the tumor microenvironment suggesting the involvement of adenosine in the resistance mechanisms observed and opening interesting perspectives for the concomitant blocking of this pathway and the PD1 / PDL-1 axis. Another strategy has been to improve the early immunomodulatory effects of TA99 mAb by combining it with oxaliplatin, a chemotherapy that promotes immunogenic death. Although the therapeutic combinations tested in this study showed encouraging in vivo effects with a significant delay in overall survival, no significant increase in the long-term anti-tumor response was observed, suggesting the establishment of other non-redundant immunosuppressive mechanisms or unsuitable combinations strategies. Both phenotypic and functional analysis of the different cellular actors of the tumor microenvironment will be a key step in the implementation of relevant combinations in association with the TA99 mAb. This work is highlighted by a phase I clinical trial (IMC-20D7S) using flanvotumab (human equivalent of mAb TA99) in 27 patients with metastatic melanoma that shows interesting clinical outcome without severe side effects, opening the way for the development of therapeutic combinations associated with this mAb.

Enterohaemorrhagic Escherichia coli O157:H7 causes haemorrhagic diarrhoea and potentially fatal renal failure in humans. Ruminants are considered to be the primary reservoir for human infection. Vaccines that reduce shedding in cattle are only partially protective and their underlying protective mechanisms are unknown. Studies investigating the response of cattle to colonisation generally focus on humoral immunity, leaving the role of cellular immunity unclear. To inform future vaccine development, the cellular immune response of cattle during EHEC O157:H7 colonisation was examined. Calves were challenged with either a phage type (PT) 21/28 strain possessing the Shiga toxin (Stx) 2a and Stx2c genes or a PT32 strain possessing the Stx2c gene only. T-helper cell associated transcripts at the terminal rectum were analysed by reverse transcriptase quantitative PCR (RT-qPCR). Induction of interferon (IFN)γ and T-bet was observed, with peak expression of both genes at 7 days in PT32 challenged calves, whilst up regulation was delayed, peaking at 21 days in PT21/28 challenged calves. Cells isolated from gastro-intestinal lymph nodes demonstrated antigen-specific proliferation and IFNγ release in response to type III secreted proteins (T3SPs); however responsiveness was suppressed in cells isolated from PT32 challenged calves. Lymph node cells showed increased expression of the proliferation marker Ki67 in CD4+ T cells from PT21/28, NK cells from PT32 and CD8+ and γδ T cells from both PT21/28 and PT32 challenged calves following ex vivo stimulation with T3SPs. Epitope mapping of rectal lymph node CD4+ T cell responses to 16 EHEC O157:H7 proteins, identified 20 CD4+ T cell epitopes specific to E. coli. The highly conserved N-terminal region of Intimin, including the signal peptide, was consistently recognised by mucosal CD4+ T cell populations from multiple animals of different major histocompatibility complex (MHC) class II haplotypes. Studies investigating the impact of secreted bacterial proteins on bovine peripheral blood mononuclear cells (PBMC) identified the ability of these proteins to cleave the surface molecule CD8 and that this phenotype was dependent on the ler virulence regulator but not the type III secretory system (T3SS) machinery. This effect was also observed in murine and ovine, but not human lymphocytes. Preliminary in vitro experiments suggest that this activity may reduce the efficiency of CD8+ T cell killing. This study demonstrates that cattle mount cellular immune responses during colonisation with EHEC O157:H7, the temporality of which is strain dependent, with further evidence of strain-specific immunomodulation.

Post-translational protein modifications (PTM) vastly increase the complexity and functional diversity of the proteome, to precisely regulate crucial cellular processes. The plant immune system is composed of complex signalling networks that are influenced by various PTMs. Activation of plant immunity is associated with a rapid burst of nitric oxide (NO), which can covalently modify cysteine thiols within target proteins by a process termed S-nitrosylation to form S-nitrosothiols (SNOs), constituting a redox-based PTM. Another key PTM involved in plant immunity is SUMOylation, an essential mechanism involving the conjugation of the small ubiquitin-like modifier (SUMO) peptide to lysine residues within target proteins. Although the targets and mechanisms of S-nitrosylation and SUMOylation are becoming evident, how these key PTMs are themselves regulated remains obscure. Work presented in this thesis reveals that during plant immune signalling, the sole Arabidopsis thaliana SUMO conjugating enzyme, SUMO CONJUGATING ENZYME 1 (SCE1), is S-nitrosylated at a highly conserved, but previously uncharacterized cysteine. S-nitrosylation of SCE1 was shown to inhibit its SUMO conjugating activity in vitro and mutational analysis revealed that the site of this modification, Cys139, is not required for enzyme activity but rather constitutes a redox-sensitive inhibitory switch. Generation and characterization of transgenic Arabidopsis plants overexpressing both wild-type and mutant forms of SCE1 revealed that Cys139 is required for efficient immunity against bacterial pathogens. Furthermore, after immune activation, S-nitrosylation of this residue inhibits global SUMOylation of proteins. These results provide evidence of a novel means of crosstalk between S-nitrosylation and SUMOylation in the context of plant immunity. The abundant cellular antioxidant, glutathione (GSH), is S-nitrosylated to form S-nitrosoglutathione (GSNO), which is thought to constitute a stable reservoir of NO bioactivity. In Arabidopsis, GSNO levels are controlled by the enzyme S-NITROSOGLUTATHIONE REDUCTASE 1 (GSNOR1), which indirectly influences the levels of protein SNOs. In this study, transgenic plants overexpressing FLAG-epitope tagged GSNOR1 were generated in various mutant backgrounds, including nitric oxide overproducer 1 (nox1), to further investigate the roles of GSNOR1 and NO in plant immunity. It was shown that ectopic GSNOR1 expression completely recovers developmental and disease susceptibility phenotypes of gsnor1, but not nox1 mutant plants, highlighting in vivo differences between accumulation of GSNO and free NO. Surprisingly, elevated NO levels in nox1 plants promote S-nitrosylation of GSNOR1, inhibiting its enzymatic activity. This suggests a previously unreported means by which NO might regulate its own bioavailability. Further work in this study revealed that recombinant GSNOR1 can be SUMOylated in vitro, which appeared to increase its enzymatic activity. Several potential SUMO modification sites were identified within GSNOR1 and mutational analysis revealed that at least one of these, Lys191, is SUMOylated. Co-immunoprecipitation experiments revealed that transgenic GSNOR1 might be SUMOylated in vivo, although the site(s) and biological function of SUMOylation were not identified. Nonetheless, these results reveal another possible layer of interplay between S-nitrosylation and SUMOylation.

Orientador: Newton Nunes / Banca: Vivian Fernanda Barbosa / Banca: Roberto Thiesen / Resumo: Avaliaram-se os efeitos de diferentes concentrações do óxido nitroso (N2O) e oxigênio, sobre os índices de lactato sérico, glicemia e características do hemograma, em suínos anestesiados com propofol e mantidos em ventilação espontânea ou controlada a pressão associada ou não à PEEP. Para isso, 48 animais machos ou fêmeas, foram distribuídos aleatoriamente em seis grupos, denominados, GN10 (FiO2 a 90% e N2O a 10%), GA10 (FiO2 a 90% e ar comprimido a 10%), GN30 (FiO2 a 70% e N2O a 30%), GA30 (FiO2 a 70% e ar comprimido a 30%), GN50 (FiO2 a 50% e N2O a 50%) e GA50 (FiO2 a 50% e ar comprimido a 50%). Empregou-se, como medicação pré-anestésica azaperona (2 mg/kg por via intramuscular), indução anestésica com propofol (dose efeito) bem como manutenção anestésica (0,5 mg/kg/min). Após a intubação o traqueotubo foi acoplado ao aparelho de anestesia inalatória para fornecimento das misturas de gases preconizadas para cada grupo. Após 100 minutos da indução anestésica, foi administrado por via intravenosa rocurônio (0,6 mg/kg IV), seguido de infusão contínua (0,6 mg/kg/hora) e inciou-se a ventilação controlada a pressão (15 cmH2O). A primeira amostra sanguínea foi coletada 20 minutos decorridos da aplicação da azaperona e imediatamente antes da indução anestésica (M0). Após 40 minutos, nova amostra de sangue foi obtida (M1), seguida de mais três colheitas decorridos 100, 175 e 220 minutos da indução da anestesia, tempos estes que coincidiram com o início da ventilação mecânica (M2), i... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study evaluated the effects of different concentrations of nitrous oxide (N2O) and oxygen, on seric level of lactate, glycemia and hematological dynamic, in propofol-anesthetized pigs under spontaneous breathing or in pressure-controlled ventilation with or without PEEP. For this, forty eight animals, males or females, were randomly divided into six groups: GN10 (FiO2 = 90% and N2O = 10%), GA10 (FiO2 = 80% and air = 10%), GN30 (FiO2 = 70% and N2O = 30%), GA30 (FiO2 = 70% and air = 30%), GN50 (FiO2 = 50% and N2O = 50%), GA50 (FiO2 = 50% and air = 50%). Azaperone was administered (2.0 mg/kg IM) as premadication. The induction was performed with propofol at sufficient doses to endotracheal intubation and for the maintenance of anesthesia, the same anesthesic were used (0.5 mg/kg/min). After 100 minutes of the induction, were administered rocuronium (0.6 mg/kg IV) followed by continuous infusion (0.6 mg/kg/h) and started mechanical ventilation (15 cmH2O). Were coleted blood samples in the first moment (M0), twenty minutes after azaperone administration. After forty minutes new blood sample were obtained (M1), followed by three more harvest past 100, 175 and 220 minutes after anesthesia induction (M2, M3 and M4 respectively). The samples were condition in microtubes with sodium fluoride or EDTA. Repeated measures analyses of variance was the statistical method used to evaluate pair wise comparisons of mean were mode, using Tukey's procedure. A P value <0,05 was considered significant. Among the main changes were between groups and moments in HT, RBC, HGB, platelets, WBC, EOS, NS, Lymph, glucose blood and lactate / Mestre

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No entanto, a prevalência de helmintos pode aumentar ou diminuir, nas dependências de fatores climáticos ou susceptibilidade do hospedeiro. Diante disso, avaliou-se a dinâmica sazonal da infecção helmíntica em equinos mantidos a pasto, a influência das diferentes características climáticas das estações do ano sobre o grau de verminose, a susceptibilidade individual e por categoria e identificar as épocas e as categorias de equinos com maior incidência de helmintoses. Foi utilizado um rebanho com 104 equinos, mestiços, classificados em diferentes categorias. Foram realizadas coletas individuais de fezes a cada 28 dias para a contagem de ovos por grama de fezes (OPG) e coproculturas para identificação de larvas. Também foram realizadas coletas de sangue para analisar possíveis alterações no hemograma dos animais associados à infecção helmíntica. Todos os animais foram pesados individualmente a cada coleta, e avaliado o escore de condição corporal, juntamente com uma avaliação clínica dos animais. Diariamente foram aferidas as temperaturas média, umidade do ar e precipitação na propriedade. Foram calculadas e apresentadas estatísticas descritivas das variáveis observadas e sua variação sequencial no tempo visando avaliar a dinâmica populacional de helmintos em equinos a pasto. As categorias que mais foram acometidas pelos helmintos foram os potros e os idosos, apresentando médias 1271,9 e 1186,5 de OPG, respectivamente. As médias de OPG encontradas nas estações da primavera, verão, outono e inverno foram, respectivamente, 1042,1; 1508,9; 817,8 e 571,1, onde todas as médias diferem significativamente entre si. Na avaliação da condição corporal foi possível observar que os animais idosos foram os que apresentaram menor ECC em todas as estações: primavera, verão, outono e inverno, sendo observado escore 3,83; 3,68; 3,56 e 3,14, respectivamente. Os resultados das coproculturas mostraram que 100% dos helmintos gastrintestinais encontrados eram da família dos pequenos estrôngilos, os Ciatostomíneos. Não foram encontradas muitas alterações nos exames hematológicos dos animais e, a maioria deles não manifestou qualquer tipo de sinais clínicos associado à doença parasitária. Foi possível concluir que a estação que os animais mais são acometidos pela verminose é no verão e que as categorias mais susceptíveis à infecção helmíntica são os potros e os animais idosos. / Parasitic infections are extremely important in horses due to the damages caused, and tend to afflict them throughout the life of the animal. However, the prevalence of helminths may increase or decrease, in the premises of climatic factors and host susceptibility. Therefore, we evaluated the seasonal dynamics of helminth infection in horses kept at pasture. a herd was used with 104 crossbred horses, classified into different categories according to age and sex. Individual fecal samples were taken every 28 days for the eggs per gram of feces (OPG) and stool cultures for identification of larvae. Also blood samples were taken to analyze possible changes in the blood count of the animals associated with helminth infection. All animals were weighed individually every collection, and rated the body condition score, along with a clinical evaluation of the animals. Daily average temperatures were measured the, humidity and rainfall on the property. Descriptive statistics of the observed variables were calculated and presented and their sequential variation in time to evaluate the population dynamics of helminths in horses grazing. The categories that were most affected by helminths were foals and elderly, with averages of OPG 1271.9 and 1186.5, respectively. The average OPG found in spring seasons, summer, autumn and winter were respectively 1042.1; 1508.9; 817.8 and 571.1, which all means differ significantly. In the evaluation of body condition was observed that the aged animals showed the lowest ECC in all seasons: spring, summer, autumn and winter, being observed score 3.83; 3.68; 3.56 and 3.14, respectively. The results of stool cultures showed that 100% of gastrointestinal helminths were found family of small strongyles the Ciatostomíneos. There were no changes in blood tests of animals and most of them did not show any clinical signs associated with the parasitic disease. It was concluded that the station that animals are more affected by worms is in the summer and that the most likely categories to helminth infection are younger foals and older animals.

O objetivo do estudo foi avaliar os efeitos da suplementação de levedura autolisada de Saccharomyces cerevisiae fornecida em duas diferentes inclusões em dietas para frangos de corte como alternativa a um antimicrobiano sobre desempenho zootécnico e avaliação do sistema imune intestinal pela realização da enumeração bacteriana, citometria de fluxo e expressão intestinal de genes ligados à resposta imune intestinal. Neste estudo foram utilizados 1260 pintos de corte machos de um dia de idade da linhagem ROSS AP95&reg; em um experimento de 1 a 35 dias de idade alojados em galpão climatizado com cama de casa de arroz nova. O experimento foi realizado em delineamento inteiramente aleatorizado, com 4 tratamentos e 7 repetições, com 45 aves por boxe. Os tratamentos utilizados foram: T1: ração basal e sem aditivo - controle negativo; T2: ração basal suplementada com 55 ppm de bacitracina de zinco - controle positivo; T3: controle negativo + 2 kg/t de levedura autolisada; T4: controle negativo + 4 kg/t de levedura autolisada. As dietas foram à base de milho e farelo de soja, sendo adicionados às rações 5% de farelo de trigo e 5% de farinha de penas e vísceras (sem tratamento prévio) com objetivo de impor um desafio nutricional aos animais. Ainda visando estimular imunologicamente os animais, aos 7 dias de idade, todas as aves foram desafiadas via ocular com uma vacina viva contendo oocistos de Eimeria sp. na dose recomendada pelo fabricante. Aos 8 dias de idade e 21 dias de idade, uma ave de cada unidade experimental, sem jejum prévio, teve sangue coletado e foi sacrificada para coleta de conteúdo intestinal ileal e cecal para realização da emumeração bacteriana de Enterococus sp., Escherichia coli e Lactobacillus sp., e para a coleta do segmento ileal para avaliar a expressão gênica intestinal de Claudin-1, IL-1&beta;, IL-4, TLR4 e MUC-2 através da PCR em tempo real. Em relação ao desempenho das aves, o tratamento T3 propiciou melhor conversão alimentar em relação a T1 até os 21 dias de idade. Para o período cumulativo, o tratamento T4 propiciou conversão alimentar semelhante ao T2, sendo esta variável melhor para estes tratamentos em relação ao controle negativo. Na enumeração de bactérias no íleo, aos 8 dias de idade, os tratamentos T3 e T4 modularam de forma distinta a contagem de Enterococus sp., e para o gênero Lactobacillus sp., ambos os grupos de levedura apresentaram menor contagem em contraste com o controle positivo. No conteúdo do ceco foi encontrado um menor número de E. coli para os animais grupo T3, diferentemente para o T2 que propiciou maior contagem. Aos 21 dias de idade, foi encontrado diferença na enumeração do gênero Enterococus sp. ileal, cuja contagem foi menor para o T2 em relação ao T1. Na na análise de citometria de fluxo, tendências foram observadas aos 8 dias de idade para o percentual de linfócitos T auxiliares (P=0,16) e para o percentual de linfócitos B (P=0,12) havendo redução com a suplementação de levedura autolisada. A mesma tendência (P=0,19) foi observada aos 21 dias de idade para a contagem de células T citóxicos. Sobre a PCR em tempo real, não foram detectadas diferenças para a expressão de Claudin-1. T2 e T4 propiciaram aumento da expressão gênica de IL-1&beta; aos 21 dias de idade em relação ao controle negativo, sendo que T2 também promoveu aumento de TLR-4 aos 8 dias de idade. Tendências foram observadas com a maior expressão de IL-4 (P=0,06) aos 21 dias de idade pelo T2 e aumento na expressão de MUC-2 (P=0,09) pelo T4 aos 8 dias de idade. Os diferentes padrões de ativação ou não de citocinas revela uma estimulação da via Th2 pelo controle positivo (aumento de IL-1&beta; e IL-4) e da via Th17 pelo tratamento suplementado com 4 kg/t de levedura autolisada (aumento de IL-1&beta;). / The objective of this study was to evaluate the effects of a Saccharomyces cerevisiae autolyzed yeast supplementation in substitution of AGP in broiler diets on performance and immune system (on two different feed inclusions for broilers diets in replacing AGP on broiler performance and evaluation of immune system trough bacterial enumeration, flux citometry and intestinal gene expression. For that, 1260 one-day-old male Ross AP95 chicks were raised from 1 to 35 days of age in a poultry house with new rice husk as litter. The experiment was arranged in a completely randomized design with 4 treatments and 7 replications, with 45 birds per pen. The treatments were: T1: basal diet and no additive - negative control; T2: basal diet supplemented with 55 ppm of zinc bacitracin - positive control; T3: negative control + 2 kg/t of autolyzed yeast; T4: negative control + 4 kg/t of autolyzed yeast. The corn-soybean meal based diets contained 5% wheat bran and 5% poultry by-product meal (with no previous treatment) in order to impose a nutritional challenge to the animals. To impose a further immunological challenge, at 7 days of age, all the birds were eye drop-vaccinated with live vaccine containing Eimeira sp. oocysts at the manufacturer recommended dosis. At 8 and 21 days of age, one chick per experimental unit, with no fasting, had the blood collected and was sacrificed for sampling the ileal and cecal intestinal contents for enumeration of Enterococus sp., Escherichia coli and Lactobacillus sp. Also, the ileal segment was sampled for intestinal gene expression of Claudin-1, IL-1&beta;, IL-4, TLR4 e MUC-2 by RNA extraction through real time PCR. For the performance results at 21 days of age, T3 had the same feed conversion rate of T1. For the cumulative grow-out, T4 had the same feed conversion rate as T2, being this variable better for the aforementioned tretaments in comparison to negative control. For ileal bacterial enumeration, at 8 days of age, T3 and T4 modulated distinctly the enumeration of Enterococus sp., and reduced the counts of Lactobacillus sp. in comparison to the positive control. In the cecal contents, the enumeration for E. coli was the lowest for T3, differing from the positive control. At 21 days of age, there was a difference in ileal Enterococus sp., with higher counts for T2 relative to T1. In the flux citometry, tendencies were observed at 8 days of age for T helper cells (P=0,16) and for B cells (P=0,12), which were reduced in the autolyzed yeast treatments. The same tendency (p=0.19) was seen at 21 days of age for T activated cytotoxic cells. For the real time PCR, there was no difference in the expression of Claudin-1 (P<0,05). T2 and T4 promoted upregulation of IL-1&beta; at 21 days of age (P<0,05) in comparison to the negative control; additionally, the antibiotic tretatment also upregulated the expression of TLR-4 at 8 days of age (P<0,05). Tendencies were observed as upregulation of IL-4 (P=0,06) at 21 days of age by positive control and upregulation of MUC-2 (P=0,09) by the treatment with 4 kg/t of autolyzed yeast at 8 days of age. The different profiles in activating or not cytokines reveals a stimulation of Th2 pathway for the positive control (upregulation of IL-1&beta; and IL-4) and Th17 pathway for the treatment supplemented with 4 kg/t of autolyzed yeast (upregulation of IL-1&beta;).

Os insetos são capazes de sobreviver em diversos ecossistemas do planeta e, mesmo estando constantemente expostos à ameaça de infecção microbiana, permanecem livres de infecções na maior parte do tempo. Essa capacidade de sobrevivência aliada à larga distribuição dos insetos em regiões totalmente diferentes tem estimulado a pesquisa de novos agentes terapêuticos nesta classe devido à descoberta de diversos componentes de mecanismos inespecíficos de combate à infecção, sendo possível sua aplicação no controle de diversas doenças. Todavia, apesar de um grande número de moléculas de defesa ter sido identificado a partir de vários insetos, pouca informação sobre suas aplicações está disponível. Desta forma, o presente trabalho elucida o perfil transcriptômico geral e dos genes de defesa do tegumento de Podalia orsilochus durante sua fase larval. Como consequência, os transcritos e os dados obtidos permitirão o auxílio em pesquisas posteriores, seja para comparação, citação, conhecimento biológico e das respostas de defesa ou das relações de filogenia do animal. / The insects are able to survive in diverse ecosystems on earth, and even being constantly exposed to the threat of microbial infections, remain free of infection for most of the time. This survivability combined with the wide distribution of insects in totally different regions has stimulated the search for new therapeutic agents in this class due to the discovery of several components of nonspecific mechanisms to fight infection, and possible implementation in the control of various diseases. However, despite a large number of defense molecules have been identified from various insects, little information is available on their applications. Thus, this paper elucidates the general transcriptomic profile and integument of defense gene Podalia orsilochus during their larval stage. As a result, the transcripts and the data obtained will aid in further research, to compare, reference, biological knowledge and defense or animal phylogeny relationships.