Distribution of 15N in amino acids during 15N-leucine infusion: impact on the estimation of endogenous flows in dairy cows

The distribution of ¹⁵N in AA during [¹⁵N]Leu infusion and its impact on the estimation of endogenous nitrogen (EN) flows in dairy cows was evaluated in 4 lactating cows equipped with ruminal, duodenal (n = 4), and ileal (n = 2) cannulae fed a silage-based diet during a 35-d experimental period. To label EN, starting on d 27, an infusion of L-[¹⁵N]Leu (0.45 mmol/h) was performed for 200 h. Samples of feed, duodenal and ileal digesta, feces, blood, urine, and mucosa of the rumen and duodenum were taken at 0900, 1100, 1300, and 1500 h on d 34 and at 0800, 1000, 1200, and 1400 h on d 35. The enrichment and fluxes of total N and individual AA were determined and used to calculate the EN flows at the duodenum, ileum, and in the feces. Based on the concept that EN comprises desquamation and secretions, EN flows were estimated, using as representative of the enrichment of EN only the enrichment of the gut mucosa (upper limit) or the average of the mucosa and the export protein enrichment (assumed to have a similar enrichment to casein; lower limit). Estimations of duodenal and fecal EN flows using the isotope dilution of ¹⁵N-total and ¹⁵N-Leu were not different and EN was an important fraction of duodenal and fecal flows, representing 14 to 30% of the duodenal flow and 18 to 31% of the fecal flow, depending on the dilution method used. The total EN flow at the duodenum is present in approximately equal proportions as either free EN or EN incorporated into bacterial protein. Ileal EN flow was 18% greater than the fecal EN flow. Using the combination of the gut and export protein, the duodenal and fecal EN flows estimated with the isotopic dilution of Leu vs. other labeled AA were less different than when estimated using the enrichment of gut mucosa alone. The current approaches have highlighted that present prediction schemes probably underestimate EN flows at the duodenum and, in consequence, overestimate net protein and AA supply. Refinement of the procedures may allow direct and accurate estimation of metabolic fecal protein, an important component of the so-called maintenance requirement of dairy cows.