Amino acid requirements of broiler breeder chickens are not well known. The indicator amino acid oxidation (IAAO) technique was adapted for use in broiler breeders as a rapid and sensitive method to determine amino acid requirements. During IAAO, phenylalanine oxidation decreases, inversely to the changes in protein synthesis, as the intake of the limiting test amino acid increases from deficient to adequate. Above the adequate level, phenylalanine oxidation remains constant. Before IAAO can be employed, the optimum priming and constant infusion doses of phenylalanine must be determined. Prelaying catheterized birds aged 20 to 24 wk were placed in closed oxidation chambers attached to a breath collection apparatus. A constant L-[1-(14)C]phenylalanine dose of 3.5 microCi/kg BW/h and priming doses of 4.5, 5.5, and 7.0 microCi/kg BW were used to determine optimal prime:constant dose ratios, minimum time taken for breath 14CO2 excretion to become constant (plateau), and adequate percentage of phenylalanine oxidized. At this constant infusion rate, the optimal priming dose of L-[1-(14)C]phenylalanine was 5.5 microCi/kg BW, resulting in a prime:constant dose ratio of 1.6:1. By using this ratio, the average time taken for breath 14CO2 to reach plateau was 60 min. Average phenylalanine oxidation at plateau, corrected for bicarbonate retention, was 5.5 +/- 1.4% (mean +/- SD), which is adequate for IAAO studies using deficient-to-excess levels of test amino acids. To the authors' knowledge, this study is the first in chickens to establish a primed, constant infusion technique using L-[1-(14)C]phenylalanine. The IAAO technique will be used in future studies to determine amino acid requirements in chickens.

The indicator amino acid oxidation technique has been widely used as a rapid and sensitive method to determine amino acid requirement. In this technique, the indicator 14C-amino acid is continuously infused and breath 14CO2 is collected to determine oxidation rate, which is used to predict requirement. However, a portion of the 14CO2 produced from amino acid oxidation is retained in the body and must be determined to correct oxidation rates based on breath 14CO2 collection. To develop the indicator amino acid oxidation technique for use in poultry, it is important to accurately measure expired and retained CO2; these measurements were accomplished using intravenous infusion of NaH14CO3. Oxidation chambers were constructed and calibrated to ensure complete collection of breath 14CO2. Hydrochloric acid was infused over 1 h into a beaker containing NaH14CO3 in four closed oxidation chambers, and the 14CO2 released was collected over 2 h. Of the initial radioactivity dose, 99.3 +/- 1.9% was recovered in collected 14CO2. We concluded that the oxidation chamber apparatus is adequate for complete recovery of expired 14CO2. In Experiment 2, we determined bicarbonate retention in five broiler breeder roosters from a primed, constant infusion of NaH14CO3 into the jugular vein for 4 h. At steady state, 86.0 +/- 1.4% of NaH14CO3 infusion was recovered in breath. Because 14% of infused 14C is retained within the body, the present results demonstrate the need to determine bicarbonate retention in chickens during oxidation studies and use it as a correction factor in subsequent experiments.