Heat processing has been used to improve protein utilization and availability of animal nutrition. However, to date, few studies exist on heat-induced protein molecular structure changes on a molecular basis. The aims of this study were to use molecular spectroscopy as a novel approach to determine heat-induced protein molecular structure changes affected by moist and dry heating and quantify protein molecular structures and nutritive value in the rumen and intestine in dairy cattle. In this study, soybean was used as a model for feed protein and was autoclaved at 120°C for 1h (moist heating) and dry heated at 120°C for 1h. The parameters assessed in this study included protein structure α-helix and β-sheet and their ratio, protein subfractions associated with protein degradation behaviors, intestinal protein availability, and energy values. The results show that heat treatments changed the protein molecular structure. Both dry and moist heating increased the amide I-to-amide II ratio. However, for the protein α-helix-to-β-sheet ratio, moist heating decreased but dry heating increased the ratio. Compared with dry heating, moist heating dramatically changed the chemical and nutrient profiles of soybean seed. It greatly decreased soluble crude protein, nonprotein nitrogen, and increased neutral detergent insoluble protein. Both dry and moist heating treatments did not alter digestible nutrients and energy values. Heating tended to decrease the nonprotein nitrogen fraction (soluble and rapidly degradable protein fraction) and true protein 1 fraction (fast-degradable protein fraction). Conversely, the true protein 3 fraction (slowly degradable fraction) significantly increased. The in situ rumen study showed that moist heating decreased protein rumen degradability and increased intestinal digestibility of rumen-undegradable protein. Compared with the raw soybeans, dry heating did not affect rumen degradability and intestinal digestibility. In conclusion, compared with dry heating, moist heating dramatically affected the nutrient profile, protein subfractions, rumen degradability, intestinal digestibility, and protein molecular structure (amide I-to-II ratio; α-helix-to-β-sheet ratio). The sensitivity of soybean seed to moist heating was much higher than that to dry heating in terms of the structure and nutrient profile changes.

1. Experiments were conducted to establish the requirements and optimal dietary ratio of lysine to threonine for fast growing male chickens (genotype Ross 308) depending on age, daily protein deposition and of dietary amino acid efficiency. 2. A total of 216 growing chickens were utilised in nitrogen-balance studies in three age periods (10 to 25 d; 30 to 45 d; 50 to 65 d) using graded levels of protein supply (60 to 360 g/kg crude protein) in lysine or threonine limiting diets. 3. Supplementation of crystalline amino acids (L-Lys, L-Thr, DL-Met and L-Arg) provided the following amino acid ratios: lysine limiting diets (Lys:Met + Cys:Thr:Arg = 1 : 1.01 : 0.91 : 1.14), threonine limiting diets (Lys : Met + Cys : Thr : Arg = 1 : 0.85 : 0.54 : 1.16). 4. The principles of the diet dilution technique using an exponential function were applied for the modelling of lysine and threonine requirements. For equal daily protein deposition, optimal lysine to threonine ratios 1 : 0.69 (10 to 25 d), 1 : 0.70 (30 to 45 d) and 1 : 0.74 (50 to 65 d) were established. 5. For the commercial growth period of fast growing chickens, the derived optimal lysine to threonine ratio was constant (1 : 0.69). The applied modelling procedure gave conclusions for quantitative requirements and optimal dietary lysine:threonine ratios in line with actual recommendations.

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N-balance studies were carried out to assess the lysine requirement of fast growing chickens (Cobb /**) at di#erent sex and age depending on crude protein deposition and e$ciency of dietary lysine utilisation. The experiments were conducted within three age periods (I : +*ῌ,/ d ; II :-*ῌ./ d ; III : /*ῌ0/ d) and 1, chickens (-0 males,-0 females) per age period. Experimental diets with six levels of graded CP content were based on high protein (HP)-soybean meal, wheat gluten and crystalline amino acids (L-Thr, DL-Met, L-Arg) in order to create lysine (..-* g Lys/+** gCP) as the first limiting dietary amino acid (constant ratio Lys : MetῌCys : Thr : Arg῍+ : +.*+ : *.3+ : +.+.). For application of a nonlinear N-utilization model, nitrogen maintenance requirement (NMR) and theoretical maximum for daily nitrogen retention (NRmaxT) were established as model parameters for further assessment of the lysine requirement depending on age, sex and daily CP-deposition. As an example, the calculated lysine requirement concentration for 0*ῌ of the theoretical potential for daily CP-deposition (+*ῌ,/ d : +.+*ῌ lysine, 0* g daily feed intake ;-*ῌ./ d : +.*-ῌ lysine, +.* g daily feed intake ; /*ῌ0/ d : *.30ῌ lysine, +1* g daily feed intake) was in close agreement with published data. However, the predicted feed intake is one of the most important factors of influence when amino acid requirement concentrations are established. The level of daily CP-deposition and the dietary amino acid e$ciency as important factors influencing the amino acid requirement data need more attention in future requirement studies.

Experiments were conducted to estimate daily N maintenance requirements (NMR) and the genetic potential for daily N deposition (ND(max)T) in fast-growing chickens depending on age and sex. In N-balance studies, 144 male and 144 female chickens (Cobb 500) were utilized in 4 consecutive age periods (I: 10 to 25 d; II: 30 to 45 d; III: 50 to 65 d; and IV: 70 to 85 d). The experimental diets contained high-protein soybean meal and crystalline amino acids as protein sources and 6 graded levels of protein supply (N1 = 6.6%; N2 = 13.0%; N3 = 19.6%; N4 = 25.1%; N5 = 31.8%; and N6 = 37.6% CP in DM). The connection between N intake and total N excretion was fitted for NMR determination by an exponential function. The average NMR value (252 mg of N/BW(kg)0.67 per d) was applied for further calculation of ND(max)T as the threshold value of the function between N intake and daily N balance. For estimating the threshold value, the principle of the Levenberg-Marquardt algorithm within the SPSS program (Version 11.5) was applied. As a theoretical maximum for ND(max)T, 3,592, 2,723, 1,702, and 1,386 mg of N/BW(kg)0.67 per d for male and 3,452, 2,604, 1,501, and 1,286 mg of N/BW(kg)0.67 per d for female fast-growing chickens (corresponding to age periods I to IV) were obtained. The determined model parameters were the precondition for modeling of the amino acid requirement based on an exponential N-utilization model and depended on performance and dietary amino acid efficiency. This procedure will be further developed and applied in the subsequent paper.