Abstract
Although stable isotope methods have been used to revisit the protein and amino acid requirements of humans in the last two decades, estimates of the minimum protein requirement of the dog have mainly been based on nitrogen balance studies. The aim of this study was: (i) to assess dog protein metabolism using the (13)C-leucine method, and (ii) to test the effects of protein deprivation and amino acid deficiency on protein metabolism. Eight dogs were fed three consecutive diets: (i) a normoprotein regimen [control; 63 g crude protein (CP)/Mcal metabolizable energy (ME)]; (ii) a protein-restricted diet (PR; 32 g CP/Mcal ME); and (iii) a protein-restricted diet that was, in addition, deficient in lysine and tryptophan (D-PR; 31 g CP/Mcal ME). The energy supply was similar for the three diets. The dogs were adapted to each diet for 2 weeks. After a 24 h fasting period, a 3 h infusion of (13)C-bicarbonate was performed, followed by a 3 h continuous infusion of L-[1-(13)C]leucine. Blood and breath samples were collected before and during the last hour of each isotope infusion for determination of plasma (13)C-alpha-ketoisocaproate and breath (13)CO(2) enrichments by mass spectrometry. Rates of protein breakdown, oxidation, and synthesis were calculated from leucine appearance into plasma, oxidation, and non- oxidative disposal, respectively, and expressed in g N/kg body weight (BW)0.75 per day, assuming body protein contains 0.08 g leucine per g protein. Protein breakdown was 3.71 +/- 0.17, 3.29 +/- 0.16 and 2.73 +/- 0.18 (mean +/- SEM) for control, PR, and D-PR, respectively (p < 0.01 D-PR versus control, and p < 0.05 D-PR versus PR). Protein synthesis was 3.08 +/- 0.13, 2.77 +/- 0.13, and 2.15 +/- 0.18 for control, PR and D-PR, respectively (p < 0.001 D-PR versus control, and p < 0.05 D-PR versus PR). Protein oxidation was 0.63 +/- 0.05, 0.53 +/- 0.05 and 0.58 +/- 0.05 for control, PR and D-PR, respectively (p=NS). These data suggest that: (i) the (13)C-leucine method can be used to assess large variations of protein turnover in dogs; (ii) dogs have the capacity to adapt their protein turnover to the level and to the quality of their protein supplies; and (iii) the dog nitrogen requirement for maintenance may be between 0.41 and 0.55 g N/kg BW(0.75) per day.
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