Made available in DSpace on 2014-07-29T15:07:32Z (GMT). No. of bitstreams: 1 Dissertacao Antonio Humberto Fleury de Melo.pdf: 1096186 bytes, checksum: a0a80798d51c0ab6d0923bc7095cea7c (MD5) Previous issue date: 2011-07-07 / Two experiments were conducted to evaluate the inclusion of levels of HMB (DL, 2-hydroxy-4-(methyl) botanical acid) methionine in diets of Nellore beef cattle finished in feedlot. In Experiment 1, 128 male, unneutered Nellore animals, with initial weight of 290 kg (± 20,50), were housed for 111 days (21 days of adaptation and 90 days of experiment) in 16 pens of 77 m2 with eight animals per pen. Isonitrogenous isocaloric diets, presenting 16% crude protein (CP) and 70% total digestible nutrients (TDN), constituted of fresh sugar cane bagasse (13.44%),soybean hulls (41.67% ), corn (18,15%), cottonseed (17,47%), soybean (5,38%), molasses (1,34%), urea (0,67%) and mineral mixture (1,88%), with four levels of HMB methionine analog inclusion were used. The treatments were T0 = control diet (no HMB), T5 = diet with 05 g HMB/animal.day-1; T10 = diet with 10 g HMB/animal.day-1; T15 = diet with 15 g HMB/animal.day-1. The experimental design was a randomized complete block, being the animals divided into blocks according to the initial weight. Final weight, total weight gain, average daily weight gain, dry matter intake and dry matter intake as % of body weight were not altered by inclusion of HMB in the diet (p>0,5). However, feed conversion (FC) (p<0,05)and feed efficiency (FE) (p<0,06) were affected by the inclusion of HMB in the diet, demonstrating that the inclusion of 10 g / animal.day-1 of HMB is the most recommended level to improve both FC and FE in cattle under these conditions. No difference was observed (p>0,05) among treatments for carcass characteristics, suggesting that under similar conditions to this experiment the inclusion of HMB in the diet does not affect the carcass quantitative characteristics. Experiment 2: was performed after adaptation to the experimentaldiet for ten days in a male Holstein animal, with approximately 200 kg of live weight with rumen cannula. Two samples of rumen fluid were collected with one week interval between each one. The samples were incubated for 0, 2, 4, 8, 12,24, 48 hours at 39°C in anaerobic conditions. A completely randomized design with four treatments and four replications was used. Diets and treatments were the same employed in Experiment 1. Fraction a (rapid degradation) of experimental diets was not altered by HMB inclusion (p>0,05). In contrast, fraction b potentially degradable) (p<0,05), fraction c (hourly rate of degradation of potentially degradable fraction) (p<0,05),Dp (potential degradability) (p<0,01) and de (effective degradability) to a passage rate (kp) at 2% per hour (p <0,01), 5% per hour (p<0.05) and 8% per hour (p<0,01) were affected by the inclusion of HMB in the diet. It was found that the inclusion of 15g/animal.day-1 of HMB in the diet positively influenced the degradability of fraction b (76,70%) and the potential degradability (90,60%). However, the effective degradability at Kp 2 and 5% h were favored for all treatments that included HMB, while at Kp 8% h, the inclusions of 5 and 10 g/animal.day-1 HMB showed higher results than other treatments (p<0,01). These results suggest that HMB has the potential to improve the degradation of high concentrate diets similar to this experiment. / Foram realizados dois experimentos com objetivo de avaliar a inclusão de níveis do análogo de metionina HMB (DL, 2-hidróxi-4-(metil) ácido botânico) em dietas de bovinos de corte da raça Nelore terminados em confinamento. Experimento 1: 128 animais da raça Nelore, macho, não castrados, peso inicial de 290 kg (±20,50), foram alojados por 111 dias (21 dias de adaptação e 90 dias de experimento) em 16 baias de 77 m2, sendo 8 animais/baia. As dietas utilizadas foram isoprotéicas e isoenergéticas, com 16% de proteína bruta (PB) e 70 % de nutrientes digestíveis totais (NDT), compostas de bagaço de cana in natura (13,44%), casca de soja (41,67%), milho (18,15%), caroço de algodão (17,47%), farelo de soja (5,38%), melaço (1,34%), uréia (0,67%) e mistura mineral (1,88%), com quatro níveis de inclusão do análogo de metionina HMB. Os tratamen tos foram T0 = dieta controle (sem HMB); T5 = dieta com 05 g HMB/animal.dia-1; T10 = dieta com 10 g HMB/animal.dia-1; T15 = dieta com 15 g HMB/animal. dia-1. O delineamento experimental foi o de blocos completos casualizados, onde os animais foram distribuídos nos blocos de acordo com o peso vivo inicial. O peso final, ganho de peso total, ganho de peso médio diário, consumo de matéria seca e o consumo de matéria seca em % do peso vivo não foram alterados pela inclusão de HMB na ração (p>0,5). No entanto a conversão alimentar (CA) (p<0,05) e a eficiência alimentar (EA) (p<0,06) foram afetadas pela inclusão de HMB nas dietas, demonstrando que a inclusão de HMB em nível de 10 g/animal.dia-1 é o mais recomendável para se obter melhora tanto na CA quanto na EA para bovinos nestas condições. Não foi observada diferença (p>0,05) entre os tratamentos para as características da carcaça, sugerindo que em condições semelhantes a este experimento a inclusão de HMB em rações não afeta as características quantitativas de carcaças. Experimento 2: foi realizado após adaptação à dieta experimental em dez dias por um animal da raça Holandês, macho, com aproximadamente 200 kg de peso vivo, munido de cânula ruminal. Foram realizadas duas coletas de líquido ruminal, com intervalo de uma semana. As amostras foram incubadas por 0, 2, 4, 8, 12, 24, 48h7 a 39ºC, em meio anaeróbio. Foi adotado delineamento inteiramente casualizado, composto de quatro tratamentos e quatro repetições. As dietas experimentais e os tratamentos são os mesmos do experimento 1. A fração a (de rápida degradação) das dietas experimentais não foi alterada pela inclusão de HMB na ração (p>0,05). Em contrapartida a fração b (potencialmente degradável) (p<0,05), a fração c (taxa horária de degradação da fração potencialmente degradável) (p<0,05), a Dp (degradabilidade potencial) (p<0,01) e a De (degradabilidade efetiva) à uma taxa de passagem (kp) de 2% por hora (p<0,01), à 5% por hora (p<0,05) e à 8% por hora (p<0,01) foram afetadas pela inclusão de HMB na dieta. Constatou-se que a inclusão de 15g/animal.dia-1 de HMB na ração influenciou positivamente a degradabilidade da fração b (76,70%) e a degradabilidade potencial (90,60%). No entanto a degradabilidade efetiva à Kp 2 e 5%h foram favorecidas para todos os tratamentos que incluíam HMB, já para Kp 8%h, as inclusões de 5 e 10 g/animal.dia de HMB foram superiores aos demais tratamentos (p<0,01). Os resultados sugerem que HMB tem potencial para ser utilizada para melhorar a degradabilidade de dietas de alto concentrado semelhante à deste experimento.

Methionine (Met) is an essential amino acid for animals and also the first limiting amino acid in a broiler diet. The dietary supplemental Met sources include the natural isoform L-methionine (L-Met), the synthetic form DL-methionine (DL-Met) and the synthetic Met precursor DL-2-hydroxy-4 (methylthio)-butanoic acid (DL-HMTBA). The objective of this dissertation was to determine the effect of different dietary Met source supplementation and Met deficiency on a series of physiological and biochemical aspects, including growth performance, global DNA methylation and methyltransferase activity, blood antioxidant profile (e.g., acute phase protein, leukocyte count), intestinal nutrient transporter gene expression, Met converting enzyme gene expression and activity, oxidative stress markers, and a potential pathway related to amino acid signaling. To achieve this goal, male Cobb-500 broilers were raised from day of hatch (d0) to d35 post-hatch and fed a diet deficient in methionine and cysteine (Met + Cys) (control) or the same diet supplemented with 0.22% DL-Met, 0.22% L-Met or 0.31% DL-HMTBA (to provide 0.22% DL-Met equivalent) to meet Met + Cys requirements. Tissues (liver, breast muscle, duodenum, jejunum and ileum) and blood samples were collected at various ages from d0 to d35 for analysis. Met supplementation significantly improved body weight gain and feed efficiency compared to the Met deficient group, but no differences were observed among DL-Met, L-Met and DL-HMTBA for growth performance parameters (P > 0.05). Met supplementation had no effect on red blood cell packed cell volume, white blood cell differential count, hepatic total DNA methylation, DNA methyltransferase and Met oxidase activity, and had limited effects on activation of p70S6K, a key amino acid signaling protein (P > 0.05). Although dietary Met sources did not change oxidative status of the treated chickens, both L-Met and DL-Met but not DL-HMTBA supplementation decreased the level of acute phase protein serum amyloid A compared to the control group (P > 0.05). The effect of Met supplementation on gene expression of nutrient transporters and Met converting enzymes were complex and dynamic. Most of the target genes demonstrated tissue- and development-dependent expression patterns, with few significant treatment effects observed. L-Met and DL-Met but not DL-HMTBA supplementation enhanced the neutral amino acid transporters ATB0,+ and B0AT gene expression in various small intestinal segments. All three Met sources increased monocarboxylic acid transporter (MCT1) gene expression in the jejunum. DL-HMTBA and L-Met fed chickens showed greater hepatic L-HMTBA oxidase (HAO1) gene expression. DL-Met increased glutamic-oxaloacetic transaminase 2 (GOT2) gene expression in the duodenum. An in vitro study with tissue explants, however, did not demonstrate a similar gene expression pattern as that in the in vivo study. Lastly, RNA sequencing results illustrated that Met deficiency could lead to many differentially expressed genes but different supplemental Met sources had no influence on hepatic gene expression profiles. In conclusion, as common dietary supplemental Met sources, L-Met, DL-Met and DL-HMTBA exhibited similarity in impacting intestinal amino acid/peptide/monocarboxylic acid transporter gene expression and Met converting enzyme activity. The regulatory roles of Met as an antioxidant and nutrient signaling in cell metabolism were not affected by different dietary supplemental Met sources. / Ph. D.