O objetivo deste trabalho foi avaliar os efeitos da inclusão de leveduras, na forma de parede celular e hidrolisada, na dieta de poedeiras sobre o desempenho produtivo, qualidade dos ovos e viabilidade econômica da suplementação destes aditivos. Dois experimentos foram realizados, para cada um deles foram alojadas 256 poedeiras distribuídas em delineamento inteiramente casualizado em 8 repetições com 8 aves. Foram avaliados os níveis de 0, 225, 450 e 900 g/t de parede celular de levedura na dieta das aves durante o período de 21 a 67 semanas de idade. Os resultados demonstraram um aumento no consumo de ração, produção e massa de ovos, além de uma maior altura do albúmen e unidade Haugh. A espessura da casca e a cor da gema também foram influenciadas pelos tratamentos. A análise da viabilidade econômica demonstrou que mesmo despendendo mais com a alimentação houve um aumento da margem bruta média, devido ao aumento da produção de ovos. Portanto, o uso da parede celular de levedura apresentou efeitos benéficos sobre o desempenho produtivo das poedeiras, como uma melhora na qualidade interna e externa dos ovos, aliado a uma maior rentabilidade da atividade. Para a levedura hidrolisada foram avaliados os níveis de 0, 1, 2 e 4 kg/t. Dois períodos foram analisados, 21 a 40 e 48 a 67 semanas de idade. A adição de levedura hidrolisada na dieta de poedeiras não apresentou melhoria das características analisadas no primeiro período, porém os benefícios surgiram a partir do segundo período. O uso da levedura além de gerar uma menor quantidade de ovos não comerciais, promoveu o aumento do consumo de ração, produção, massa e peso dos ovos, melhoria na qualidade dos ovos em relação a unidade Haugh, altura do albúmen, espessura e resistência da casca; e ainda melhorou a conversão alimentar por massa e dúzia de ovos. A suplementação de levedura hidrolisada para poedeiras proporcionou incremento da margem bruta total média em todos os níveis testados, demonstrando-se viável economicamente. A inclusão de levedura hidrolisada na dieta de poedeiras de 48 a 67 semanas de idade exerceu efeito maximizador do desempenho produtivo e qualidade de ovos, sendo economicamente viável seu uso. / The purpose of this study was to evaluate the effects of including yeast in the form of cell wall and hydrolyzed, in the diet of laying hens on production performance, egg quality and economic viability of these feed additives. Two experiments were conducted, for each of them 256 laying hens were housed and distributed in a completely randomized design with 8 repetitions of 8 birds. The level of 0, 225, 450 and 900 g/t of yeast cell wall were evaluated in the diet of birds from 21 to 67 weeks of age. The results showed an increase in feed intake, egg production and mass as well as a higher albumen height and Haugh unit. The shell thickness and yolk color were also affected by treatments. The economic viability analysis demonstrated that even spending more on feed an increase in the average gross margin due to increased egg production was observed. Therefore, the cell wall yeast showed a beneficial effect on production performance of laying hens, represented by an improvement of internal and external egg quality, combined with higher profitability. Hydrolyzed yeast was evaluated at the levels of 0, 1, 2 and 4 kg/t. Two periods were analyzed, 21-40 and 48-67 weeks of age. The addition of hydrolyzed yeast in the diet of laying hens did not show improvement of the analyzed characteristics in the first period, however the benefits occurred in the second period. The use of yeast although generate a minor amount of non-commercial eggs, promoted an increase in feed consumption, production, egg mass and weight, improving quality of eggs in relation to Haugh unit, albumen height, shell thickness and resistance; and also improves the feed conversion by eggs mass and dozen. The supplementation of hydrolyzed yeast for laying hens provided increase overall average gross margin on all tested levels, demonstrating to be economically viable. The inclusion of hydrolyzed yeast in the diet of laying hens from 48-67 weeks of age maximized production performance and egg quality, being its use economically viable.

Submitted by ANGELICA AUGUSTA GRIGOLI DOMINATO null (angelica@unoeste.br) on 2017-02-16T18:51:51Z No. of bitstreams: 1 Angélica A. Grigoli Dominato.pdf: 4485568 bytes, checksum: 4abc8fe39b8c64a5ed8be37f2be077de (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-02-20T20:26:27Z (GMT) No. of bitstreams: 1 dominato_aag_dr_sjrp.pdf: 4485568 bytes, checksum: 4abc8fe39b8c64a5ed8be37f2be077de (MD5) / Made available in DSpace on 2017-02-20T20:26:27Z (GMT). No. of bitstreams: 1 dominato_aag_dr_sjrp.pdf: 4485568 bytes, checksum: 4abc8fe39b8c64a5ed8be37f2be077de (MD5) Previous issue date: 2017-01-20 / A atividade metabólica fúngica, especialmente nos endofíticos, favorece a secreção de moléculas como antibióticos, pigmentos, enzimas e polissacarídeos, que podem ser aplicadas nas indústrias alimentícia, cosmética, farmacêutica, entre outras. A diversidade química dos exopolissacarídeos (EPS) bem como a possibilidade de sua utilização como substrato para diferentes modificações químicas (carboxilação, sulfatação e metilação) aumentam seu espectro de aplicação. Realizar o cultivo submerso dos fungos endofíticos Neofusicoccum parvum, Fusarium sp e Colletotrichum gloeosporioides para a produção de EPS foi o primeiro objetivo deste trabalho. Uma vez obtidos, os EPS foram purificados e quimicamente caracterizados. Sulfatação e ensaio da atividade anticoagulante foram realizados. Os parâmetros concentração de inóculo e tempo de cultivo foram estabelecidos para maior produção dos EPS, por fermentação submersa. Etapas de purificação, por centrifugação, foram efetuadas após análises por cromatografia de exclusão estérica a alta pressão, com detecção por índice de refração (HPSEC/RID). Os EPS purificados [precipitado do C. gloeosporioides (C. gloeosporioidesprec) e os três sobrenadantes] mostraram-se praticamente isentos de proteínas. A hidrólise ácida e subsequente análise dos hidrolisados por cromatografia de troca aniônica a alta pressão com detecção por amperometria pulsada (HPAEC/PAD) indicaram que apenas o C. gloeosporioidesprec era uma glucana. A análise por cromatografia gasosa acoplada à espectrometria de massa (GC/MS) mostrou um único derivado, 1,3,5 tri-O-acetil, 2,4,6-tri-O-metil glucitol com fragmentos de massa (m/z 118, 161, 203, 234.1) condizente com uma glucana do tipo (1→3). Os espectros de FT-IR apresentaram sinais na região de impressão digital, 926 cm-1 e 820 cm-1, típicos de polissacarídeos em configuração . Esses resultados foram confirmados por ressonância magnética nuclear bidimensional (HSQC), com um único acoplamento C1/H1, em 99,3/5,18 ppm e um sinal deslocado para campo baixo, 82,8/3,74 ppm, característico de C-3 substituído, indicando que o EPS é uma α-(1→3)-glucana. A sulfatação desta molécula resultou em α-(1→3)-D-glucanasulf com DS de 0,75 que foi utilizada nos ensaios de atividade anticoagulante. O aumento do tempo para a coagulação, nos testes do APTT (Tempo de Tromboplastina Parcial Ativada) e TT (Tempo de Trombina), foi concentração-dependente, indicando que [α-(1→3)-D-glucanasulf] pode atuar como um inibidor da via intrínseca da coagulação sanguínea e da conversão do fibrinogênio em fibrina, caracterizando-a como um potencial anticoagulante. / Fungal metabolic activity, especially in the endophytic, favors secretion of molecules such as antibiotics, pigments, enzymes and polysaccharides, which can be applicable by food, cosmetic and pharmaceutical industries, and others. The chemical diversity of the exopolysaccharides (EPS), as well as the possibility of their use as substrate for different chemical modifications (carboxylation, sulfation and methylation) increases their application spectrum. Submerged cultivation of the endophytic fungi Neofusicoccum parvum, Fusarium sp and Colletotrichum gloeosporioides for the production of EPS was the first aim of this study. Once the EPS were obtained, they were purified and chemically characterized. Chemical modification by sulfation and anticoagulant activity assays were performed. Cultivation to obtain EPS were performed in submerged fermentation. The inoculum concentration and incubation time parameters were set in order to obtain a higher amount of EPS. Purification by centrifugation was performed after analysis by high pressure steric exclusion chromatography with refractive index detection (HPSEC / RID). Purified EPS [precipitate of C. gloeosporioides (C. gloeosporioidesprec) and the three supernatants] proved to be virtually free of protein polysaccharides. Acid hydrolysis and subsequent analysis of the hydrolysates with high performance anionic exchange chromatography with amperometric detection (HPAEC/PAD) indicated that only the C. gloeosporioidesprec was a glucan. Analysis from gas chromatography and mass spectrometry showed a single derivative, 1,3,5-tri-Oacetyl, 2,4,6-tri-O-methyl glucitol with mass fragments (m/z 118, 161, 203, 234.1) consistent with a (1→3)-glucan. FT-IR spectra showed absorption bands typical from carbohydrate and signals in the digital region at 926 cm-1 and 820 cm-1, typical of polysaccharides in the α- configuration. These results were confirmed by two-dimensional nuclear magnetic resonance (HSQC), with a single C1/H1, in 99.2/4.98 ppm, typical of one α bonding, and low-field 82.6/3.55 ppm signal displacement, characteristic of substituted C-3, indicating that EPS is an α-(1→3)-glucan. Sulfation of this molecule resulted in α- (1→3)-glucansulf with DS of 0.75 which was used in the anticoagulant activity assays. The increase in coagulation reaction time in the APTT (Activated Partial Thromboplastin Time) and TT (Thrombin Time) tests was concentration-dependent, indicating that [α-(1→3)-D-glucansulf] might act as an inhibitor of the intrinsic via of blood clotting and conversion of fibrinogen into fibrin, characterizing it as a potential anticoagulant.

O objetivo deste trabalho foi avaliar os efeitos da inclusão de leveduras, na forma de parede celular e hidrolisada, na dieta de poedeiras sobre o desempenho produtivo, qualidade dos ovos e viabilidade econômica da suplementação destes aditivos. Dois experimentos foram realizados, para cada um deles foram alojadas 256 poedeiras distribuídas em delineamento inteiramente casualizado em 8 repetições com 8 aves. Foram avaliados os níveis de 0, 225, 450 e 900 g/t de parede celular de levedura na dieta das aves durante o período de 21 a 67 semanas de idade. Os resultados demonstraram um aumento no consumo de ração, produção e massa de ovos, além de uma maior altura do albúmen e unidade Haugh. A espessura da casca e a cor da gema também foram influenciadas pelos tratamentos. A análise da viabilidade econômica demonstrou que mesmo despendendo mais com a alimentação houve um aumento da margem bruta média, devido ao aumento da produção de ovos. Portanto, o uso da parede celular de levedura apresentou efeitos benéficos sobre o desempenho produtivo das poedeiras, como uma melhora na qualidade interna e externa dos ovos, aliado a uma maior rentabilidade da atividade. Para a levedura hidrolisada foram avaliados os níveis de 0, 1, 2 e 4 kg/t. Dois períodos foram analisados, 21 a 40 e 48 a 67 semanas de idade. A adição de levedura hidrolisada na dieta de poedeiras não apresentou melhoria das características analisadas no primeiro período, porém os benefícios surgiram a partir do segundo período. O uso da levedura além de gerar uma menor quantidade de ovos não comerciais, promoveu o aumento do consumo de ração, produção, massa e peso dos ovos, melhoria na qualidade dos ovos em relação a unidade Haugh, altura do albúmen, espessura e resistência da casca; e ainda melhorou a conversão alimentar por massa e dúzia de ovos. A suplementação de levedura hidrolisada para poedeiras proporcionou incremento da margem bruta total média em todos os níveis testados, demonstrando-se viável economicamente. A inclusão de levedura hidrolisada na dieta de poedeiras de 48 a 67 semanas de idade exerceu efeito maximizador do desempenho produtivo e qualidade de ovos, sendo economicamente viável seu uso. / The purpose of this study was to evaluate the effects of including yeast in the form of cell wall and hydrolyzed, in the diet of laying hens on production performance, egg quality and economic viability of these feed additives. Two experiments were conducted, for each of them 256 laying hens were housed and distributed in a completely randomized design with 8 repetitions of 8 birds. The level of 0, 225, 450 and 900 g/t of yeast cell wall were evaluated in the diet of birds from 21 to 67 weeks of age. The results showed an increase in feed intake, egg production and mass as well as a higher albumen height and Haugh unit. The shell thickness and yolk color were also affected by treatments. The economic viability analysis demonstrated that even spending more on feed an increase in the average gross margin due to increased egg production was observed. Therefore, the cell wall yeast showed a beneficial effect on production performance of laying hens, represented by an improvement of internal and external egg quality, combined with higher profitability. Hydrolyzed yeast was evaluated at the levels of 0, 1, 2 and 4 kg/t. Two periods were analyzed, 21-40 and 48-67 weeks of age. The addition of hydrolyzed yeast in the diet of laying hens did not show improvement of the analyzed characteristics in the first period, however the benefits occurred in the second period. The use of yeast although generate a minor amount of non-commercial eggs, promoted an increase in feed consumption, production, egg mass and weight, improving quality of eggs in relation to Haugh unit, albumen height, shell thickness and resistance; and also improves the feed conversion by eggs mass and dozen. The supplementation of hydrolyzed yeast for laying hens provided increase overall average gross margin on all tested levels, demonstrating to be economically viable. The inclusion of hydrolyzed yeast in the diet of laying hens from 48-67 weeks of age maximized production performance and egg quality, being its use economically viable.

The present study investigated the expression profiles of nine genes involved in immune defense of the Pacific white shrimp Litopenaeus vannamei and their responses to oral administration of beta-1,3-glucan. The nine immune related genes were beta-glucan binding protein-high density lipoprotein (BGBP-HDL), lipopolysaccharide/beta-glucan binding protein (LGBP), hemocyanin, prophenoloxidase (proPO), transglutaminase (TGase), penaeidin-3 (PEN-3), crustin, cytosolic manganese superoxide dismutase (cMnSOD), and lysozyme. A series of experiments were carried out in the study including: (1) cDNA cloning and characterization of proPO and LGBP; (2) tissue mRNA expressions of the nine genes in adult shrimp; (3) expression and localization of the nine genes during larval and postlarval ontogenic development; (4) the effects of dietary beta-1,3-glucan on the expression of the nine genes. The cDNA cloning study showed that the proPO cDNA contains an open reading frame of 2061 bp and encodes a 686 amino-acid peptide. The protein sequence of the proPO has a similarity of 85% with those of Penaeus monodon and P. semisulcatus and has an identity of between 58 and 77% with other crustaceans. Northern blot analysis revealed that proPO was constitutively expressed mainly in hemocytes. Its transcripts were observed in hemocytes and many other tissues when detected with RT-PCR. The results of in situ hybridizations showed that the hemocytes that infiltrated in tissues were responsible for the positive signals. The LGBP cDNA contains an open reading frame of 1104 bp and encodes a 367 amino-acid protein with a 17 a. a. signal peptide. The protein sequence of the LGBP has a similarity of 97% with LGBP of L. stylirostris, >90% identity with BGBP of P. monodon and LGBP of Fenneropenaeus chinensis and has an identity of between 63 and 86% with other crustaceans. Northern blot analysis revealed that LGBP was constitutively expressed mainly in hepatopancreas. The results of in situ hybridizations showed that the hepatopancreatic F cells might be the major cell type for LGBP production. Using the complete cDNAs of proPO and LGBP and partial fragments of the other seven genes, their tissue expressions were analyzed by conventional RT-PCR, quantitative real time PCR (qPCR) and in situ hybridization. The results demonstrated that BGBP-HDL, LGBP and hemocyanin were mainly expressed in the hepatopancreas and their expressions levels were about 10 to 30% those of

Lafora disease (LD) is a rare yet invariably fatal form of epilepsy characterized by progressive degeneration of the central nervous and motor systems and accumulation of insoluble glucans within cells. LD results from mutation of either the phosphatase laforin, an enzyme that dephosphorylates cellular glycogen, or the E3 ubiquitin ligase malin, the binding partner of laforin. Currently, there are no therapeutic options for LD, or reported methods by which the specific activity of glucan phosphatases such as laforin can be easily measured. To facilitate our translational studies, we developed an assay with which the glucan phosphatase activity of laforin as well as emerging members of the glucan phosphatase family can be characterized. We then adapted this assay for the detection of endogenous laforin activity from human and mouse tissue. This laforin bioassay will prove useful in the detection of functional laforin in LD patient tissue following the application of therapies to LD patients. We subsequently developed an in vitro readthrough reporter system in order to assess the efficacy of aminoglycosides in the readthrough of laforin and malin nonsense mutations. We found that although several laforin and malin nonsense mutations exhibited significant drug-induced readthrough, the location of the epitope tag used to detect readthrough products dramatically affected our readthrough results. Cell lines established from LD patients with nonsense mutations are thus required to accurately assess the efficacy of aminoglycosides as a therapeutic option for LD. Using hydrogen-deuterium exchange mass spectrometry (DXMS), we then gained insight into the molecular etiology of several point mutations in laforin that cause LD. We identified a novel motif in the phosphatase domain of laforin that shares homology with glycosyl hydrolases (GH) and appears to play a role in the interaction of laforin with glucans. We studied the impact of the Y294N and P301L LD mutations within this GH motif on glucan binding. Surprisingly, these mutations did not reduce glucan binding as expected, rather enhancing the binding of laforin to glucans. These findings elucidate the mechanism by which laforin interacts with and acts upon glucan substrates, providing a target for the development of therapeutic compounds.

Orientador: Aulus Cavalieri Carciofi / Banca: Helio Jose Montassier / Banca: Iracilda Zeppone Carlos / Resumo: Vários derivados da parede celular da levedura Saccharomyces cerevisiae conhecidamente agem sobre a imunidade, no entanto a ação, especialmente da fração beta-glucano, foi pouco demonstrada em cães. Para o estudo dos possíveis efeitos sobre a imunidade na espécie canina foram empregadas quatro dietas isonutrientes, contendo uma fonte de parede celular de levedura (PCL), duas fontes de beta-glucano (BG1 e BG2) e uma dieta controle (CT). Foram utilizados 24 cães da raça beagle, adultos, divididos em quatro grupos de seis animais. As dietas foram fornecidas por um período total de 126 dias e as avaliações incluíram hemograma e avaliações bioquímicas, dosagem de anticorpos anti-Leptospira, imunofenotipagem de linfócitos sanguíneos, avaliação da concentração de IgA em fezes, teste de hipersensibilidade cutânea tardia e dosagens de citocinas em sobrenadante de cultura celular. Os animais foram submetidos a desafio antigênico com vacina contra leptospirose no dia 42. Os dados foram avaliados pelo procedimento GLM do SAS, sendo as médias comparadas pelo teste de Tukey (p≤0,1). Nos exames bioquímicos houve discreta variação entre os tratamentos e ao longo dos dias. No hemograma notou-se aumento dos linfócitos para BG2. A dosagem de anticorpos anti-Leptospira, mostrou baixos títulos, não havendo boa resposta à vacinação. A imunofenotipagem revelou um aumento dos linfócitos T totais, T helper, T citotóxicos e linfócitos B no grupo BG2 e de linfócitos T citotóxicos e linfócitos B para o grupo PCL. Apesar da variação da concentração de IgA fecal ao longo dos dias, os tratamentos não influenciaram tais parâmetros. O teste de hipersensibilidade cutânea tardia mostrou um aumento na resposta à inoculação da vacina, para os grupos PCL e BG2. Na dosagem de citocinas em sobrenadante de cultura celular, apenas foi observada diferença na quantificação de TNF-α, ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Some products from the cell wall of the yeast Saccharomyces cerevisiae are known to act on the immunity, however this action, especially of the beta-glucan fraction, has never been demonstrated in dogs. To study these effects on the immunity of dogs four isonutrient diets were made, containing one source of yeast cell wall (YCW), two sources of beta-glucan (BG1 and BG2) and a control diet (CT). 24 adult beagle dogs were used, divided in four groups of six animals. Diets were given for a 126 days period. Evaluations included complete blood count and biochemistry profile, quantification of antibodies against Leptospira, immunophenotyping of blood lymphocytes, IgA concentration in feces, delayed-type hypersensitivity test and quantification of cytokines in cell culture supernatant. Animals were exposed to antigen challenge, by the vaccine against leptospirosis on day 42. Data were analyzed by the GLM procedure of the SAS software and the means were compared by the Tukey test (p<0,01). Biochemistry profile showed slight differences among the groups. A increase in lymphocyte count was observed for BG2 treatment. Quantification of antibodies against Leptospira showed low titles, with poor response to vaccination. Immunophenotyping revealed an increase during the time in total T cells, helper and cytotoxic T cells, and B lymphocytes for BG2 and of cytotoxic T cells and B lymphocytes for YCW group. Despite the variation in fecal IgA concentration during the time, treatments did not influence these parameters. Delayed-type hypersensitivity test showed an increased response to the vaccine inoculation, for YCW and BG2 groups. In the quantification of cytokines in cell culture supernatant the only difference observed was in TNF-α concentration, being BG2 higher than CT. We concluded that both yeast cell wall and beta-glucan fraction act on dogs' immunity / Mestre

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The traditional use of antibiotics in aquaculture as growth promoters has been limited due to the negative effects caused by these drugs. As an alternative to the use of these drugs has been sought manipulation of the microbiota of the gastrointestinal tract of aquatic animals through the use of oligosaccharides and dietary fibers with prebiotic potential. Thus, this study aimed to apply different methodologies to obtain Dietary Fiber Concentrates (DFC) = mucilage (MG); pectin (PN) and β-glucan + manan (βG + M) and evaluate the prebiotic potential of these supplements in the diet of juvenile jundiá (Rhamdia quelen). The determination of the nutritional composition of the ingredients revealed that the predominant component in all DFCs were dietary fiber and insoluble fiber. The DFC that had higher extraction yield was βG + M (19.81 ± 8.54%), followed by pectin (14.54% ± 2.72), and mucilage (7.18 ± 1.54%). The composition of mucilage and pectin had a greater diversity of monosaccharides, since the βG+M consisted primarily of mannose (74.5%) and glucose (24.3%). The supplementation of DFC in jundiás diet was assessed for eight weeks through study of growth, body nutrient deposition, digestive enzymes, biochemical and metabolic parameters, responses to stress and immune and intestinal morphology. The jundiás supplemented with DFCs achieved higher growth than the control group and similar to animals supplemented with 5 g kg-1 commercial prebiotic (CP 5). Most somatic parameters and whole fish proximate composition were influenced by supplementation of DFCs. The supplementation of pectin promoted lower activity of digestive enzymes in relation the control group. The animals supplemented with DFC obtained positive changes in biochemical parameters. Furthermore, jundiás showed no response to application of the stressor, maintaining basal cortisol levels. The fish supplemented with DFCs had higher hepatic glycogen stores in relation the control group. Moreover, supplementation with DFCs increased the height of intestinal villi of jundiá. However, these values were lower for the animals of the group PC 5. For thickness of the epithelium this variable was higher in the control group compared to animals supplemented with β-glucan+Manana. / O uso tradicional de antibióticos na aquicultura como promotores de crescimento tem sido limitado em função dos efeitos negativos promovidos por estes medicamentos. Como alternativa ao uso destas drogas, tem se buscado a manipulação da microbiota do trato gastrointestinal dos animais aquáticos através da utilização de oligossacarídeos e de fibras alimentares com potencial prebiótico. Neste sentido, este estudo teve como objetivo aplicar metodologias para obtenção de diferentes Concentrados de Fibras Alimentares (CFAs) = Mucilagem (MG); Pectina (PN) e β-Glicana+Mananas (βG+M) e avaliar o potencial prebiótico destes suplementos na dieta de juvenis de jundiá (Rhamdia quelen). A determinação da composição nutricional dos ingredientes revelou que os componentes predominantes em todos os CFAs obtidos foram fibra alimentar total e fibra insolúvel. O CFA que apresentou maior rentabilidade de extração foi a βG+M (19,81%±8,54), seguida da Pectina (14,54%±2,72) e Mucilagem (7,18%±1,54). A composição da Mucilagem e Pectina obtiveram maior diversidade de monossacarídeos, já a βG+M consistiu basicamente de manose (74,5%) e glicose (24,3%). A suplementação dos CFAs na dieta de jundiás foi avaliada durante oito semanas, através de estudo de crescimento, deposição corporal de nutrientes, enzimas digestivas, parâmetros bioquímicos e metabólicos, resposta ao estresse e imunológica e morfometria intestinal. Os jundiás suplementados com os CFAs obtiveram crescimento superior em relação ao grupo controle e similar aos animais suplementados com 5 g kg-1 de prebiótico comercial (PC 5). A maioria dos parâmetros somáticos e de composição centesimal de peixe inteiro foram influenciados pela suplementação dos CFAs. A suplementação de Pectina promoveu menor atividade das enzimas digestivas em relação ao grupo controle. Os animais suplementados com os CFAs obtiveram alterações positivas nos parâmetros bioquímicos avaliados. Além disso, os jundiás não mostraram resposta à aplicação do agente estressor, mantendo os níveis de cortisol basal. Os peixes suplementados com os CFAs obtiveram maiores estoques de glicogênio hepático em relação ao grupo controle. Além do mais, a suplementação com os CFAs promoveu aumento na altura de vilos intestinais dos jundiás. Porém, estes valores foram menores em relação aos animais do grupo PC 5. Para espessura do epitélio (EE) esta variável foi maior no grupo Controle comparado aos animais suplementados com β- glicana + Manana.

Made available in DSpace on 2014-06-11T19:23:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-23Bitstream added on 2014-06-13T20:11:36Z : No. of bitstreams: 1 zaine_l_me_jabo.pdf: 606188 bytes, checksum: da0b30ee696bae955544945c1be515d9 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Biorigin Sa / Vários derivados da parede celular da levedura Saccharomyces cerevisiae conhecidamente agem sobre a imunidade, no entanto a ação, especialmente da fração beta-glucano, foi pouco demonstrada em cães. Para o estudo dos possíveis efeitos sobre a imunidade na espécie canina foram empregadas quatro dietas isonutrientes, contendo uma fonte de parede celular de levedura (PCL), duas fontes de beta-glucano (BG1 e BG2) e uma dieta controle (CT). Foram utilizados 24 cães da raça beagle, adultos, divididos em quatro grupos de seis animais. As dietas foram fornecidas por um período total de 126 dias e as avaliações incluíram hemograma e avaliações bioquímicas, dosagem de anticorpos anti-Leptospira, imunofenotipagem de linfócitos sanguíneos, avaliação da concentração de IgA em fezes, teste de hipersensibilidade cutânea tardia e dosagens de citocinas em sobrenadante de cultura celular. Os animais foram submetidos a desafio antigênico com vacina contra leptospirose no dia 42. Os dados foram avaliados pelo procedimento GLM do SAS, sendo as médias comparadas pelo teste de Tukey (p≤0,1). Nos exames bioquímicos houve discreta variação entre os tratamentos e ao longo dos dias. No hemograma notou-se aumento dos linfócitos para BG2. A dosagem de anticorpos anti-Leptospira, mostrou baixos títulos, não havendo boa resposta à vacinação. A imunofenotipagem revelou um aumento dos linfócitos T totais, T helper, T citotóxicos e linfócitos B no grupo BG2 e de linfócitos T citotóxicos e linfócitos B para o grupo PCL. Apesar da variação da concentração de IgA fecal ao longo dos dias, os tratamentos não influenciaram tais parâmetros. O teste de hipersensibilidade cutânea tardia mostrou um aumento na resposta à inoculação da vacina, para os grupos PCL e BG2. Na dosagem de citocinas em sobrenadante de cultura celular, apenas foi observada diferença na quantificação de TNF-α,... / Some products from the cell wall of the yeast Saccharomyces cerevisiae are known to act on the immunity, however this action, especially of the beta-glucan fraction, has never been demonstrated in dogs. To study these effects on the immunity of dogs four isonutrient diets were made, containing one source of yeast cell wall (YCW), two sources of beta-glucan (BG1 and BG2) and a control diet (CT). 24 adult beagle dogs were used, divided in four groups of six animals. Diets were given for a 126 days period. Evaluations included complete blood count and biochemistry profile, quantification of antibodies against Leptospira, immunophenotyping of blood lymphocytes, IgA concentration in feces, delayed-type hypersensitivity test and quantification of cytokines in cell culture supernatant. Animals were exposed to antigen challenge, by the vaccine against leptospirosis on day 42. Data were analyzed by the GLM procedure of the SAS software and the means were compared by the Tukey test (p<0,01). Biochemistry profile showed slight differences among the groups. A increase in lymphocyte count was observed for BG2 treatment. Quantification of antibodies against Leptospira showed low titles, with poor response to vaccination. Immunophenotyping revealed an increase during the time in total T cells, helper and cytotoxic T cells, and B lymphocytes for BG2 and of cytotoxic T cells and B lymphocytes for YCW group. Despite the variation in fecal IgA concentration during the time, treatments did not influence these parameters. Delayed-type hypersensitivity test showed an increased response to the vaccine inoculation, for YCW and BG2 groups. In the quantification of cytokines in cell culture supernatant the only difference observed was in TNF-α concentration, being BG2 higher than CT. We concluded that both yeast cell wall and beta-glucan fraction act on dogs` immunity

Made available in DSpace on 2014-12-17T14:16:36Z (GMT). No. of bitstreams: 1 KarinaMM_DISSERT.pdf: 2256022 bytes, checksum: e9ccee900e81515e9367adddbe68fd65 (MD5) Previous issue date: 2012-02-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The frequency of disseminated candidiasis caused by yeast has enhancing in intensive care unit. Despite the availability of new antifungal drugs, C. albicans sepsis mortality causes can be as high as 30-40%. So, it has been needed to looking for a new therapeutic medicament that helps in treatment and prevention of this infection. Previous data that demonstrated that particulated ?-glucan stimulates the immune system and experiments of this work were conducted to investigating if ?-glucan extracted from Saccharomices cerevisiae, could modified the evolution of mouse model C. albicans systemic infection. Balb/c mice with sepsis and ?-1,3 glucan treated or not were analyzed the influence of ?-1,3 glucan in survival of the animals, in the fungal burdens in kidney, in the production of urea and TNF even in the histopathology of kidney. The experiments shown that the infected animals a nd glucan treated had great survival (p<0,05), less unit form colony in kidney and normal levels of urea. In the kidney histopathology of not glucan treated animals it has seen more lesions when compared with treated animals. So we conclude that ?-1,3 glucan could stimulate the immune system against disseminated C. albicans / A frequ?ncia de candid?ase invasiva causada por leveduras tem aumentado nas unidades de terapia intensiva. Apesar dos importantes avan?os no desenvolvimento de agentes antif?ngicos, a sepse por Candida ssp tem inaceit?vel atribui??o a mortalidade de 40-50% de pessoas. Existe a necessidade de se identificar novos agentes terap?uticos que auxiliem no tratamento e preven??o dessa infec??o. Assim, o objetivo deste trabalho foi avaliar se a ?-1,3 glucana particulada, extra?da do Saccharomyces cerevisiae, seria capaz de modificar a evolu??o da infec??o sist?mica por Candida albicans em modelo experimental. Camundongos Balb/c submetidos ? sepse e tratados ou n?o com ?-1,3 glucana particulada foram avaliados com rela??o ao tempo de sobrevida, quantifica??o das unidades formadoras de col?nia nos rins, dosagem de ur?ia e TNF s?ricos e an?lise histopatol?gica renal para avalia??o de infiltrado inflamat?rio e presen?a do fungo no tecido renal. Os resultados mostraram que os animais infectados por Candida albicans e tratados com glucana apresentaram uma maior sobrevida (p<0,05), uma menor quantidade de unidades formadoras de col?nia nos rins e valores de ur?ia normais. Na an?lise dos resultados da histopatologia foi observado um maior dano no tecido renal dos animais pertencentes ao grupo n?o tratado com glucana, diferentemente do observado nos animais tratados pela glucana os quais apresentaram pequeno dano renal. Assim, podemos concluir que a glucana foi capaz de estimular o sistema imune na defesa contra infec??o disseminada por C. albicans

Oats is a crop that contains a high amount of fiber, protein and fat, but like all other crops it contains mostly starch. In this study the focus has been oat flours and brans with different b-glucan content. The health benefits of b-glucan, a soluble fiber are well documented and a correlation between intake of b-glucan with high molecular weight and a low glycemic response has been observed. Food with a low glycemic index can lower the risk for diseases like type 2 diabetes, cardiovascular diseases and obesity. Also a connection between intake of b-glucan with high molecular weight and a reduction of LDL-cholesterol has been observed. b-glucans from oat absorb water and build a viscous gel, which make them an interesting component when developing new products, as a fat replacer in for example meat products and pastries. To optimize the use of flours and brans with a modified b-glucan content in new applications, the water absorption was measured with a method called Solvent Retention Capacity and the viscosity with a Rapid Viscosity Analyzer (RVA). The results showed that a higher amount of b-glucan in the flour or bran, a higher water holding capacity (WHC) was observed. The WHC for oat flour with a b-glucan content at 2% was calculated to 73±7%, while the WHC for oat bran with a b-glucan content at 28%, was calculated to a WHC of 880±45%. A comparison of different flours and brans indicates that dietary fiber, where b-glucan have the greatest impact on the WHC. The result from the RVA indicates that a flour with a combination of a high b-glucan content (0.24g) and high starch content (3.72g) leads to a high viscosity 12700 cP, compared to other flours or brans with either a lower b-glucan content (0.12g) or lower starch content (0.12g) gives lower final viscosity, 5390 and 780 cP. The result also indicates that other factors such as a smaller particle size and a higher temperature during the heating step (95°C instead of 64°C) might give a higher viscosity.