1. The study aimed to establish the optimum time after ingestion and optimum sampling site for the development of an ileal amino acid digestibility assay for broiler chickens. 2. To establish the optimal sampling time, 4-week-old broiler chickens were given one of 6 protein sources (meat-and-bone, soyabean, cottonseed, fish, maize and wheat meals) as the sole source of protein in a test diet. The diets contained chromic oxide as an indigestible marker. The birds were starved for 24 h, fed and subsequently killed for sampling of ileal digesta (terminal 15 cm) at 2, 3, 4, 5 and 6 h after the start of feeding. 3. For the soyabean, fish, wheat and maize meal diets, sampling time had no significant effect on apparent ileal nitrogen digestibility, whereas for the meat-and-bone and cottonseed meal diets there was a significant quadratic effect of sampling time. The amount of digesta collected was maximised and the mean apparent ileal nitrogen digestibility had the lowest variation around the 4 h sampling time. 4. To establish the optimum sampling site, 4-week-old chickens were given either a meat-and-bone, a soyabean or a wheat bran meal-based diet. The birds were killed 4 h after the start of feeding, and digesta were sampled from 0-10, 0-15, 0-20 or 0-25 cm of terminal ileum. 5. There was no significant effect of sampling site on the apparent ileal digestibility of dietary nitrogen. The terminal 15 cm of ileum was considered a preferred site for sampling ileal digesta from broiler chickens.
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