O presente trabalho teve como objetivo avaliar o desempenho de novilhas da raça Nelore, suplementadas com níveis de proteína bruta em pastagem de capimbraquiarão (Brachiaria brizantha cv. Marandu), no período de transição água-seca. O experimento foi conduzido na Faculdade de Zootecnia e Engenharia de Alimentos - FZEA/USP em Pirassununga-SP. Utilizou-se quatro tratamentos: Controle (CTRL) sendo sal mineral; Suplemento com 20% de proteína bruta (20%); suplemento com 40% de proteína bruta (BC 40%), sendo esses de baixo consumo e suplemento 40% de proteína bruta de alto consumo (AC 40%). O ciclo de pastejo foi de 35 dias, sendo sete dias de ocupação e 28 dias de descanso. O delineamento experimental foi em blocos completos casualizados com 4 repetições.Foram utilizadas 96 novilhas da raça Nelore com peso médio inicial de 316 kg e cerca de 18 meses de idade, num período experimental de 105 dias, tendo início em 09 de abril e término em 27 de agosto de 2008, considerado período de transição água-seca. Para o ganho de peso médio diário, não houve diferença estatística entre os tratamentos AC 40% e BC 20% (p<0,05), sendo o GPD de 0,470 e 0,369 gramas/dia, respectivamente, enquanto que para os tratamentos CTRL, 20% e BC 40%, não houve diferença estatística (P>0,05), sendo osGPD de 0,336, 0,369 e 0,353 gramas/dia, respectivamente. A utilização de suplementos protéicos no desempenho de novilhas teve desempenhos satisfatórios no período de transição água-seca, porém, para a recomendação da utilização, recomenda-se realizar uma análise econômica, pois somente assim, garantirá qual será o melhor tratamento para o período. / The aim of this study was to evaluate the performance of Nelore heifers supplemented with crude protein levels in pasture of Brachiaria grass (Brachiaria brizantha cv. Marandu), during the transition period of wet-dry seasons. The experiment was conducted at the Faculty of Animal Science and Food Engineering - FZEA / USP in Pirassununga-SP. It was used four treatments: Control (CTRL) being mineral salt, supplement with 20% of crude protein (20%) supplement with 40% of crude protein (BC 40%), as low consumption and with crude protein of 40% as high consumption (AC 40%). The grazing cycle was 35 days, seven days of occupation and 28 days of interval. A randomized complete block design, with 4 repetitions was utilized. It was used 96 Nelore heifers with an average weight of 316 kg and around 18 months, in a probationary period of 105 days, starting on 09 April and ending in August 27, 2008, considered the transitional period of wet-dry seasons. For the average daily gain, there was no statistical difference between treatments AC 40% and 20% (p<0,05), being GPD of 0.470 and 0.369 grams/day, respectively, while for the treatments CTRL, 20% and BC 40%, there was no statistical difference (P>0,05), being GPD of 0.336, 0.369 and 0.353 grams/day, respectively. The use of crude protein supplements on performance of heifers had performed satisfactorily during the transition period of wet-dry seasons however, to recommend the use, it is suggest to perform an economic analysis, for only thus ensure the best treatment for the period.

O estudo dos teores protéicos oferecidos aos animais possui crucial importância, pois estes exercem influência determinante no desenvolvimento da infecção parasitária. Investigou-se o efeito da utilização de dois diferentes teores protéicos (18,5% e 8,6% de PB na MS) sobre as alterações fisiopatológicas desenvolvidas por bezerros primo infectados com o nematóide intestinal Cooperia punctata. Objetivando determinar se a suplementação possibilitaria aos animais infectados responderem mais prontamente ao desafio parasitário, realizaram-se estudos de composição corporal, retenção de compostos nitrogenados, acompanhamento clínico completo, avaliação de peso e carcaça, avaliação quantitativa e qualitativa da carga parasitária e exames histopatológicos. Os animais que receberam suplementação apresentaram um quadro clínico menos severo, perderam menos peso, tiveram um maior rendimento de carcaça, apresentaram lesões teciduais mais exuberantes, maiores taxas de excreção urinária de nitrogênio, expulsaram os parasitos para porções distais do trato gastrointestinal e demonstraram uma eficiência de retenção de nitrogênio 3,6 vezes maior que os animais mantidos na dieta basal. Demonstrou-se que a suplementação protéica resultou em efeitos positivos para os animais infectados. / The study of protein levels offered to cattle is very important when one aims at nematode control, since they can effectively affect the development of parasitic infection. The effect of two different levels - 18.5% and 8.6% CP on DM basis on the pathophysiology of Cooperia punctata infection on naive calves was examined. Aiming at determine if protein supplementation allow animals to better withstand parasitic challenge, several parameters were evaluated: body composition, nitrogen retention, clinical evolution, weight gain, carcass yeld, quantitative and qualitative assessment of parasitic burden and histopathology. Supplemented animal presented a milder clinical signs, lost less weight, had higher carcass yeld, greater tecidual lesions, higher urinary nitrogen excretion, expelled parasites to distal parts of gastrointestinal tract and showed a 3.6 greater efficiency for nitrogen retention than animal kept on basal diet. It became clear that protein supplementation resulted in benefits to infected animals.

Title: Effect of protein (amino acid) ingestion on muscle protein synthesis following resistance exercise. Purpose: The main objective of this thesis is to verify the three basic factors of the amount, type and timing of protein intake based on scientific studies and literature, to provide the most objective and accurate information and procedure on the methodology of nutrition and supplementation associated with the intake of protein / amino acids after strength training and how it all affects muscle synthesis. Summery: The theoretical part of the thesis, discusses the factors affecting muscle protein synthesis, which stimulate growth and tissue regeneration, based on optimal stress response. Logically, it starts from general, i.e. the explanation of terms such as muscle tissue, the stimulation of muscle tissue and its manifestations and changes, nutrition factors and muscle stimulation, the mechanism of dietary factors (proteins/amino acids), specificity of protein/ amino acids in their application to answer the three key issues, which are summarized in the section named scientific studies, which focuses on the effect of intake of protein/ amino acids, in relationship to the efficiency of protein synthesis after strength training. The section summarizes, in detail, the questions of timing,...

Orientador: Flávio Dutra de Resende / Banca: Ricardo Andrade Reis / Banca: Marco Antonio Alves Balsalobre / Resumo: Objetivou-se verificar o efeito da suplementação de baixo consumo, sobre o desempenho de novilhos Nelore mantidos em pastagem de Brachiaria brizantha cv. Marandu. O período experimental foi de julho de 2008 a março de 2009 e foi divido em duas fases, secas e águas. Os tratamentos durante o período da seca constituíramse de um controle, sal com uréia, e suplemento mineral enriquecido, este que era acrescido de proteína verdadeira e aditivos. Já no período das águas avaliou-se um tratamento controle, sal mineral e suplemento mineral enriquecido. O delineamento experimental utilizado foi inteiramente casualizado, em esquema de parcela subdividida. No caso da avaliação conjunta das duas fases o delineamento utilizado foi inteiramente casualizado, em esquema fatorial 2 x 2, sendo os fatores os tipos de suplementação em cada uma das fases. Realizaram-se avaliações de pasto e pesagem dos animais a cada ciclo de avaliação dentro dos períodos estudados. No período da seca foi observada diferença estatística em função dos tratamentos avaliados somente no terceiro ciclo. Os animais que receberam suplementação mineral enriquecida (proteína mais aditivos) ganharam 0,600 kg/dia, contra 0,517 kg/dia daqueles receberam sal com uréia. No primeiro e segundo ciclo os tratamentos não apresentaram diferença significativa em relação ao ganho, 0,307 e 0,113 kg/dia. No período das águas os animais que receberam suplementação mineral enriquecida ganharam mais peso do que os animais que receberam suplementação mineral, 0,800 e 0,696 kg/ dia. Concluiu-se que a suplementação mineral acrescida de proteína e aditivos, permite incrementos no ganho de peso dos animais mantidos em regime de pastejo na ordem de 15% / Abstract: The objective was to test the effect of low intake supplementation on performance of Nelore steers grazing Brachiaria brizantha cv. Marandu. The experiment was from July 2008 to March 2009 divided into two phases, wet and dry seasons. The treatments during the dry season consisted of a control, a salt urea, and mineral supplement, that it was increased with true protein and additives. In the wet season we evaluate a control treatment, mineral salt and a supplement mineral enriched. The lineation experiment was completely randomized in parcel scheme subdivided. In the case of joint evaluation of the two phases, the lineation was completely randomized in scheme factorial 2X 2, the factors being the type of supplementation in each phase. Evaluations were performed on grazing and animals weighed each evaluation cycle within the periods studied. During the dry season was a statistical difference in the treatments evaluated only in the third cycle. The animals that received mineral supplementation enriched (protein more additive) gained 0.600 kg/day, against 0.517 kg/day those who received salt with urea. In the first and second cycle the treatments showed no significant difference in relation to gain, 0.307 e 0,113 kg/day. In wet season the animals receiving mineral supplementation enriched gained more weight than animals receiving just mineral supplementation, 0.800 e 0.696 kg/day. Concludes the mineral supplementation enriched with protein and additive, allows greater weight gains than animals kept in grazing to order 15% / Mestre

Title: Effect of protein (amino acid) ingestion on muscle protein synthesis following resistance exercise. Purpose: The main objective of this thesis is to verify the three basic factors of the amount, type and timing of protein intake based on scientific studies and literature, to provide the most objective and accurate information and procedure on the methodology of nutrition and supplementation associated with the intake of protein / amino acids after strength training and how it all affects muscle synthesis. Summery: The theoretical part of the thesis, discusses the factors affecting muscle protein synthesis, which stimulate growth and tissue regeneration, based on optimal stress response. Logically, it starts from general, i.e. the explanation of terms such as muscle tissue, the stimulation of muscle tissue and its manifestations and changes, nutrition factors and muscle stimulation, the mechanism of dietary factors (proteins/amino acids), specificity of protein/ amino acids in their application to answer the three key issues, which are summarized in the section named scientific studies, which focuses on the effect of intake of protein/ amino acids, in relationship to the efficiency of protein synthesis after strength training. The section summarizes, in detail, the questions of timing,...

Made available in DSpace on 2015-03-26T13:55:24Z (GMT). No. of bitstreams: 1 texto completo.pdf: 614632 bytes, checksum: eac726a0efd1eec87578c9745a474e81 (MD5) Previous issue date: 2014-02-26 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / This work aimed to evaluate the effects of ruminal and/or abomasal protein supplementation in cattle fed tropical forages on nutritional variables, degradation of insoluble fiber, activity of fibrolytic enzymes and rumen bacterial communities profile. Two experiments were carried out sequentially, and in the same way, differing only on the forage quality that was defined based on crude protein content: medium quality (784 g of crude protein/ kg of dry matter) and high quality (986 g of crude protein/ kg of dry matter). Four Nellore young bulls, averaging 227±11 kg body weight and fitted with ruminal and abomasal cannullas, were used. The following treatments were evaluated: control (only forage), ruminal nitrogenous compounds supplementation (daily supply of 230 g of crude protein in the rumen), abomasal nitrogenous compounds supplementation (daily supply of 230 g of crude protein in the abomasum), and ruminal and abomasal nitrogenous compounds supplementation (daily supply of 230 g of protein, being 115 g in the rumen and 115 g in the abomasum). Casein was used as protein source for supplementation and basal diet consisted of Tifton 85 (Cynodon spp) hay. Two 4 x 4 Latin squares experimental trials, balanced for residual effects, with four treatments, four animals and four experimental periods lasting 29 days each, were implemented sequentially, one for each forage as stated above. There were no effects (P>0.05) of supplementation scheme or forage quality on organic matter (OM) intake. On the other hand, neutral detergent fiber (NDF) intake was higher (P<0.05) when animals were fed high quality forage. The total digestibility of OM was increased (P<0.05) by supplementation and by increasing forage quality. The digestibility coefficient of NDF was not affected by supplementation scheme (P>0.05), but was increased by the enhancing in forage quality. The apparent ruminal nitrogen balance (RNB) was increased (P<0.05) when animals were supplemented in rumen or in both rumen and abomasum cannula. It is worth highlighting that no supplementation or exclusively abomasal supplementation led to negative values of RNB. The apparent nitrogen balance (NB) was increased (P>0.05) when high quality forage was fed to the animals and by supplementation, no matter if the supplement was infused into rumen, abomasum or both. Ruminal supplementation enhanced (P<0.05) the concentration of rumen ammonia nitrogen (RAN). Although, animals supplemented in rumen and abomasum or exclusively in abomasum did not exhibit increase in RAN concentrations (P>0.05) when compared to no supplemented animals. The bacterial richness and diversity (Shannon-Wiener index) associated with the solid phase of rumen digesta were not affected (P>0.05). However, richness and diversity associated with the liquid phase were increased (P<0.05) by the improvement in forage quality. NDF degradation rate (kp) was enhanced (P<0.05) by ruminal supplementation. Carboximetilcellulase activity was decreased (P<0.05) by the abomasal and ruminal/abomasal supplementation. Xylanase activity was decreased (P<0.05) by both post-ruminal supplementation schemes, when high quality forage was fed to the animals. It was not detected correlations among variables associated with bacterial diversity and activity and variables associated with nutritional performance, excepted for the positive correlation between kp and RNB Clustering analyses based on the Unweighted Pair Group Method (UPGMA) evidenced that distinct bacterial communities are associated with liquid and solid fraction, independently of the forage quality. Clustering analyses also showed that forage quality affects the diversity of liquid phase associated bacteria, even though, effects on solid phase was not clear. None of the evaluations related to the microbial community were able to detect clear effects of supplementation on the microbial community diversity. Improving forage quality enhances fiber digestibility, nitrogen balance and modifies the bacterial community associated with liquid phase. On the other hand, protein supplementation enhances the nitrogen retention in animal body. / Objetivou-se avaliar os efeitos da suplementação proteica ruminal e/ou abomasal em bovinos consumindo forragens tropicais sobre as características nutricionais, a atividade enzimática fibrolítica, a degradação in vitro da fibra insolúvel e o perfil da comunidade bacteriana ruminal. Dois experimentos foram realizados sequencialmente diferindo apenas na qualidade do feno ofertado, sendo de média qualidade (78,4 g de proteína bruta/kg de matéria seca) e alta qualidade (98,6 g de proteína bruta/kg de matéria seca), respectivamente. Os procedimentos experimentais foram os mesmos para ambas as forragens. Foram utilizados quatro novilhos Nelore, não castrados, com peso corporal médio de 227±11 kg, fistulados no rúmen e no abomaso. Foram avaliados os seguintes esquemas de suplementação: controle (somente forragem); suplementação nitrogenada ruminal (fornecimento diário de 230 g de proteína suplementar no rúmen); suplementação nitrogenada abomasal (fornecimento diário de 230 g de proteína suplementar no abomaso) e suplementação nitrogenada ruminal e abomasal (fornecimento diário de 230 g de proteína suplementar, sendo 115 g no rúmen e 115 g no abomaso). A alimentação volumosa basal foi constituída por feno de tifton 85 (Cynodon spp) e como fonte de compostos nitrogenados suplementares foi utilizada a caseína (caseína pura, Labsynth). O experimento foi conduzido segundo delineamento em quadrado latino 4 x 4 balanceado para efeitos residuais, sendo dois quadrados sequenciais, um para cada forragem, com quatro esquemas de suplementação, quatro animais e quatro períodos experimentais com 29 dias em cada quadrado. Não foram observados efeitos (P>0,05) da qualidade da foragem ou do esquema de suplementação sobre o consumo de matéria orgânica (MO). Por outro lado, o consumo de fibra em detergente neutro (FDN) foi superior (P<0,05) quando forragem de alta qualidade foi ofertada aos animais. O coeficiente de digestibilidade total da MO foi incrementado (P>0,05) pela suplementação e pela melhoria na qualidade da forragem. O coeficiente de digestibilidade da FDN não foi afetado pelo esquema de suplementação (P>0,05), mas foi superior quando os animais receberam forragem de alta qualidade (P<0,05). O balanço aparente de compostos nitrogenados no rúmen (BNR) foi ampliado (P<0,05) pela suplementação no rúmen e no rúmen-abomaso. Ressalta-se que a ausência de suplementação ou a suplementação exclusiva no abomaso proporcionam valores negativos de BNR. O balaço aparente de compostos nitrogenados no organismo animal foi ampliado (P<0,05) pelo fornecimento de suplemento, independentemente do local de suplementação, e pelo fornecimento de forragem de melhor qualidade A suplementação no rúmen incrementou (P<0,05) a concentração de nitrogênio amoniacal ruminal (NAR). Por outro lado, animais recebendo suplementação no rúmen e abomaso ou exclusivamente no abomaso apresentaram concentrações de NAR similares (P>0,05) a animais não suplementados. A riqueza e a diversidade microbiana da fase sólida da digesta ruminal não foram afetadas (P>0,05). Entretanto, a riqueza e a diversidade bacteriana associada à fase líquida da digesta ruminal foram incrementadas (P<0,05) pela melhoria na qualidade da forragem. A taxa de degradação da FDN foi ampliada (P<0,05) pela suplementação no rúmen. A atividade da carboximetilcelulase foi reduzida (P<0,05) pela suplementação no abomaso e rúmen-abomaso. A atividade da xilanase foi deprimida (P<0,05) pela suplementação pós-ruminal somente quando forragem de alta qualidade foi fornecida aos animais. Não foi observada correlação (P>0,05) entre as variáveis associadas à atividade enzimática e diversidade microbiana e as características ligadas ao desempenho nutricional, exceção feita à correlação positiva (P<0,05) entre a taxa de degradação da FDN e o BNR. A avaliação multivariada da diversidade permitiu evidenciar que comunidades bacterianas distintas estão associadas às frações líquida e sólida, independente da qualidade da forragem ofertada. Percebeu- se que a variação na qualidade da forragem influencia a diversidade microbiana associada à fração líquida do fluido ruminal, embora não tenha exercido tal efeito sobre a fração sólida. Em nenhuma avaliação percebeu-se efeito claro do esquema de suplementação sobre o perfil da comunidade microbiana ruminal. A melhoria da qualidade da forragem amplia a digestibilidade da fibrae a retenção de nitrogênio e é capaz de modificar a comunidade bacteriana associada à fração liquida do conteúdo ruminal. A suplementação proteica, por sua vez, independentemente do local de suplementação, amplia a retenção de nitrogênio no organismo animal.

Capillary electrophoresis mass spectrometry (CE-MS) is a versatile instrumental method for metabolomics, which allows for comprehensive metabolite profiling of volume-limited biological specimens in order to better understand the molecular mechanisms associated with chronic diseases, including an alarming epidemic of obesity worldwide. Multiplexed CE separations enable high-throughput metabolite screening with quality assurance to prevent false discoveries when combined with rigorous method validation, robust experimental designs, complementary statistical methods, and high-resolution tandem mass spectrometry (MS/MS) for unknown metabolite identification. In this thesis, multiplexed CE-MS technology is applied for both targeted and untargeted metabolite profiling of various biological fluids, including covalently bound thiol-protein conjugates, as well as free circulating metabolites in serum and plasma, and excreted/bio-transformed compounds in urine due to complex host-gut microflora co-metabolism. This work was applied to characterize aberrant metabolic responses of obese subjects in response to dietary challenges, and measure the benefits of dietary interventions that reduce adiposity without deleterious muscle loss. Chapter 2 presents, a simple, sensitive yet robust analytical protocol to expand metabolome coverage in CE-MS for the discovery of labile protein thiols in human plasma using a rapid chemical derivatization method based on N-tert-butylmaleimide (NTBM). Chapter 3 describes targeted metabolite profiling of serum and plasma to investigate the differential metabolic responses between healthy and unhealthy obese individuals before and after consumption of a standardized high-caloric meal, respectively. Chapter 4 of this thesis describes an untargeted metabolite profiling strategy for urine using multisegment-injection (MSI)-CE-MS for elucidating the effects of protein supplementation following a short-term dietary weight-loss intervention study. This work revealed six urinary metabolites that were classified as top-ranking treatment response biomarkers useful for discriminating between subjects consuming carbohydrate (control), soy, and whey supplemented diets. In summary, this thesis demonstrated the successful implementation of multiplexed CE-MS technology for biomarker discovery in nutritional-based metabolomic studies as required for more effective treatment and prevention of obesity for innovations in public health. / Thesis / Doctor of Philosophy (PhD)