THESIS-ABSTRACT Supplementation levels of exogenous alpha-amylase in broilers diets.

This study aimed to evaluate the supplementation levels of an exogenous alpha-amylase in broilers diets and compare two indicators in determining the diets energy. The experiment was divided into two parallel evaluations, being one of performance and the other of metabolism. In performance assay, 1,700 one-day-old Cobb-500 male chicks were used. The animals were distributed in 50 experimental plots and evaluated five treatments with ten replicates in a completely randomized design (CRD). The treatments were: a positive control (PC), a negative control (NC) and three alpha-amylase supplementation levels 200, 400 and 600 g/t, and the NC was formulated with 50 and 90 kcal of energy reduction in relation to the PC to the phases from 1 to 21 days and from 22 to 42 days, respectively. In the metabolism assay were used 240 animals, 150 birds for stage from 14 to 21 days and 90 birds to stage from 35 to 42 days of age and the treatments were the same as the performance assay, with six replicates per treatment in CRD. All diets of metabolism test contained the digestibility indicators Lipe® (eucalyptus purified lignin) and chromic oxide (Cr2O3), in concentrations of 0.05 and 1.0%, respectively. In the period from 1 to 21 days old, no significant differences were observed in weight gain (WG) (P > 0.05), however, feed intake (FI) was found higher by using 200 ppm of enzyme (P < 0.05) and better feed conversion (FC) with the PC (P < 0.05). From 22 to 42 days, no significant differences were observed on the WG (P > 0.05), but were observed lower FI and better FC to PC treatment (P < 0.05). In the period from 1 to 42 days of age, significant differences were also not observed on the WG (P > 0.05), but there was lower FI and better FC for the PC treatment (P < 0.05). The AMEn (apparent metabolizable energy corrected for nitrogen balance), determined using the total collection, reaffirmed the values ​​calculated for the PC and NC with intermediate data obtained from the enzyme use (200, 400 and 600 ppm). Comparing the total collection using Lipe® and Cr2O3, a correlation was observed only for the PC results, that were always higher, and for the NC results, that were lower for the three methodologies. For IDE (ileal digestible energy), determined by Cr2O3, significant differences were observed (P < 0.05) and it presented higher values in the PC treatment and lower values in the NC. In IDE determining by Lipe®, significant was observed (P < 0.05), showing higher value in the PC treatment. It is concluded that in the metabolism assessments, exogenous alpha-amylase tested was effective in increasing the metabolizable energy (ME), however, not enough to be equivalent to the ME of PC treatment, a result that was better expressed in the total collection, AMEn by Lipe® and IDE by Lipe® methodologies. The performance results reflect the superiority of PC treatment, pointing enzyme limitation to supply the energy deficit practiced in this work.

Thesis Abstract Levels and forms of vitamin D in broilers diets.

This study aimed to evaluate the concentration effects of two vitamin D isoforms, cholecalciferol (D3) and 25-hydroxycholecalciferol (25-OHD3) in broilers diets on performance, bone and physiological features of these birds. Of a total of 1920 one-day-old male chicks Cobb-500 were used from commercial hatchery, reared under bed creation systems. The animals were distributed in six treatments and eight replicates with 40 birds per treatment in a completely randomized design. The following vitamin D supplementation levels were applied: 70 and 87.5 µg/kg feed in initial phase; 56 and 70 µg/kg feed during the growth phase, and 35 and 47.35 µg/kg of feed in final phase of creation, obtained from two forms (D3 and 25-OHD3). The treatments consisted of supplementation of two levels from each isolated source and their associations (60% D3 + 40% 25-OHD3) according to the study phases. In the metabolism assay, 480 birds (14 and 35 days of age) were separated to be used for evaluation of calcium (Ca) and phosphorus (P) retention and excretion during the periods of 19 to 21 days and 40 to 42 days of age. The diets were based on corn and soybean meal, with supplementation of phytase (500 FTU/kg). The performance, bone characteristics, plasma levels, bone radiographic density, carcass yield, and P and Ca retention were evaluated. In the initial creation phase, we observed an increased P excretion by broilers fed diets supplemented with vitamin D3 (P < 0.05). In addition, the association between the two vitamin D isoforms resulted in higher retention of Ca and P than the birds fed diets supplemented only with vitamin D3 (P < 0.05), and higher P retention when compared to birds fed diets supplemented with 25-OHD3 (P < 0.05). Dietary supplemental 25-OHD3 at 87.5 µg/kg resulted in higher plasma levels of Ca in relation to the same supplemented source with 70 µg/kg at 21 days of age (P < 0.05). In the final phase, the birds fed diets supplemented with vitamin D3 presented the lowest values of consumption and retention of Ca and P (P < 0.05). The association between the forms (D3 and 25-OHD3) reduced the excretion values of Ca and P (p < 0.05). The birds receiving a source of 25-OHD3 and the association had better feed conversion ratio and higher bone ash content (P < 0.05). At all stages studied D3 + 25-OHD3 combined supplementation increased tibial density of broilers in relation to supplementation of only vitamin D3 (P < 0.05). Results of this study indicate that the addition of 25-OHD3 in the feed supplemented with vitamin D3 improve the feed conversion, increase the Ca plasma levels, and also increases bone density, providing higher retention coefficients of Ca and P and lower P excretion, regardless of the development phase of these birds.

Thesis Abstract Bean (Phaseolus vulgaris L.) lines: chemical composition and protein digestibility.

The bean represents the main source of proteins for the low income populations, although the digestibility of those proteins is relatively low. Consequently, the programs of plant genetic breeding have been working on the search for new lines with higher protein levels. Thus, with the purpose of supplying information to the researchers, in this study, 21 bean (Phaseolus vulgaris L.) lines were analyzed for the centesimal and mineral composition, protein digestibility, phenolic compounds, and trypsin inhibitor. The entirely randomized experimental design was used with 21 treatments (lines) and three repetitions. All values were within the following ranges: 22.34 to 36.28 g crude protein/100 g dry matter (DM); 7.56 to 20.91 g neutral detergent fiber/100 g DM; 0.53 to 2.55 g fat/100 g DM and 2.97 to 4.87 g ashes/100 g DM. The levels of phosphorus, potassium, calcium, magnesium, and sulfur, in g/100 g DM, varied from 0.45 to 0.72; 1.51 to 2.48; 0.03 to 0.28; 0.18 to 0.34 and 0.28 to 0.45, respectively. Regarding copper, manganese, zinc and iron, the levels, in mg/kg DM, varied from 11.37 to 17.73; 14.93 to 28.90; 36.67 to 69.90 and 71.37 to 126.90, respectively. The in vitro protein digestibility varied from 18.03 to 48.32%. The levels of phenolic compounds varied from 0.28 to 1.08 mg acid tanic/100 g DM and the one of trypsin inhibitor from 59.93 to 151.07 trypsin inhibited units/mg DM. Among the lines with higher protein contents, "ESAL 569" (beige with brown stripe) presented the largest protein digestibility and considerable levels of minerals. "P-180" (beige with brown stripe) was one of the lines with higher crude protein contents and digestibilities, and also presented high levels for most of the minerals. No relation between protein digestibility and the contents of phenolic compounds or trypsin inhibitor was observed.

Technical note: Evaluation of markers for estimating duodenal digesta flow and ruminal digestibility: Acid detergent fiber, sulfuric acid detergent lignin, and n-alkanes.

The amount of digesta flowing to the duodenum is a relevant measurement for the evaluation of nutrient supply to ruminants, which is usually estimated in animals fitted with a duodenal T-type cannula using internal or external markers. This study evaluated acid detergent fiber (ADF) compared with external (C32n-alkane) and internal [sulfuric acid lignin (ADL) and n-alkanes C31 and C33] markers for estimating duodenal flow and(or) ruminal digestibility of dry matter (DM) in cattle and sheep. In the first assay, 4 duodenally cannulated Holstein steers housed in metabolism cages, dosed with C32n-alkane, and fed Avena strigosa plus concentrate and increasing levels of tannin extract to reduce ruminal digestibility, were used in a Latin square design. The mobile-bag technique was used to measure the intestinal disappearance of ADL and ADF from forage (Avena strigosa, Pennisetum purpureum, Cynodon dactylon, and Medicago sativa) and concentrate (corn grain, soybean meal, and sunflower meal) samples that were previously incubated in the rumen of additional fistulated steer for 12, 24, 36, or 48 h. The ADF concentration in residues recovered in the feces was strongly related to the ADF concentration in residues at the duodenum (R(2)=0.93, standard deviation=30.0, n=901). This relationship showed a lower precision for ADL fraction (R(2)=0.88, standard deviation=12.6, n=590). In a second assay, duodenal flow and ruminal DM digestibility were calculated from the duodenal and fecal concentration of either marker. We observed a significant effect of marker type on ruminal DM digestibility values, and the effect of tannin treatments was observed only when ADF or ADL was used as the marker. The lowest residual error was obtained for ADF. Ruminal DM digestibility was, on average, higher for C31 and C(33)n-alkanes, and the use of dosed C(32)n-alkane resulted in a negative value. In the third assay, a data set of 235 individual observations was compiled from digestibility trials to compare ADF and ADL as markers for estimating duodenal digesta flow in wethers (n=204) and cattle (n=31). We observed a strong relationship between markers (R(2)=0.84 in sheep and R(2)=0.88 in cattle), but variance analysis within trials indicated that ADF was more precise than ADL. In conclusion, in digestibility trials in which fecal output was measured and spot samples of the duodenal digesta were obtained, duodenal flow and ruminal digestibility of the DM may be estimated from the relationship between the ADF concentration in feces and that in the duodenal digesta of ruminants.

Avaliação de métodos laboratoriais para estimar a digestibilidade e o valor energético de dietas para ruminantes / Evaluation of laboratorial methods to estimate the digestibility and energetic value of ruminant diets

Foi avaliada a acurácia dos métodos laboratoriais para estimar a digestibilidade e o valor energético de dietas para bovinos de corte. As dietas experimentais foram isonitrogenadas e compostas por silagem de milho e 25, 40, 55 ou 70 por cento de concentrado. Os valores de digestibilidade medidos in vivo foram comparados com os obtidos nos ensaios in situ, in vitro e in vitro/gases, e com valores estimados a partir de equações matemáticas baseadas na composição química das dietas. O ensaio in vivo foi realizado com quatro bovinos em delineamento em quadrado latino 4×4. Quatro animais foram usados para o ensaio in situ e quatro corridas foram realizadas para os ensaios in vitro. Não houve interação significativa de dietas versus métodos. As taxas de degradação calculadas a partir dos ensaios in situ e in vitro/gases não foram acuradas para estimar o valor nutritivo dos alimentos, e o método in vitro foi o que melhor estimou a digestibilidade das dietas. A equação de Weiss superestimou o valor nutricional das dietas por superestimar a digestibilidade da fibra e por subestimar a excreção endógena fecal, mas essa equação foi mais acurada que as equações de McDowell.

The accuracy of laboratorial methods to estimate the digestibility and energetic value of beef cattle diets was evaluated. Experimental diets were isonitrogenous and composed by corn silage, and 25, 40, 55, or 70 percent of concentrate. Digestibility values measured in vivo were compared to those obtained in situ, in vitro, and in vitro/gases assays, as well to values estimated from mathematical equations based on the chemical composition of diets. The in vivo assay was carried out using four cattle in a Latin Square experimental design. Four animals were used for in situ assay and four runs were carried out for in vitro assays. Significant interaction of method vs. diet was not observed. As single indicators, degradation rates calculated from in situ and in vitro/gas assays were not accurate to estimate the nutritive value of feeds. Among all tested methods, the in vitro assay was the most accurate to estimate the digestibility of diets. Weiss equation overestimated the nutritional value of diets by overestimated fiber digestibility and by underestimated fecal endogenous excretion. However, it was more accurate than McDowell equations.