The simultaneous estimation of the amounts of protozoal, bacterial and dietary nitrogen entering the duodenum of steers.

Four steers were given straw and tapioca diets, twice daily, in a 4 X 4 Latin-square design. These diets, containing 4.2 g nitrogen/kg dry matter (DM), were further supplemented with either urea, decorticated groundnut meal (DCGM), untreated (UT) casein or formaldehyde-treated (FT) casein to give a total of 19.7 g N/kg DM and 10.5 MJ/kg DM daily. Concurrent samples of rumen bacteria and protozoa and abomasal digesta were collected for each period of the experiment and the concentrations of 2-aminoethyl phosphonic acid (AEPA), diaminopimelic acid (DAPA), total nitrogen (TN), total phosphorus (TP), amino acids and hexosamines were determined in the dried preparations. The nature of the dietary supplements had little effect on the concentrations of most of these constituents or on the total protozoal numbers. Abomasal digesta samples marked with polyethylene glycol (PEG) and chromic oxide for flow estimation were collected over 24 h, and the proportions of protozoal-N, bacterial-N and microbial-N estimated simultaneously using the markers AEPA, DAPA and RNA respectively. These digesta-N components were also estimated using an amino acid profiling (AAP) method which gave, in addition, estimates of the dietary and endogenous components. For the diets containing casein, the proportion of dietary casein was estimated directly using casein-P as a marker. Estimates of the respective mean proportions of microbial-N in abomasal digesta non-ammonia-N (NAN) for the diets containing urea, DCGM, UT casein or FT casein were: AEPA 0.56, 0.32, 0.27 and 0.16; DAPA 0.88, 0.70, 0.81 and 0.57; RNA 0.98, 0.85, 0.92 and 0.53. Giving FT casein significantly (P less than 0.001) increased the flow of casein-N at the abomasum and a significantly (P less than 0.001) greater proportion of casein-N was found in abomasal NAN (0.51 v. 0.09) where FT rather than UT casein was given. The AAP method gave results for the proportions of microbial- and dietary-N (where casein was given) which were, in general, slightly lower than those obtained using RNA and casein-P as markers. Agreement with estimates of bacterial protein (from DAPA) and of protozoal protein (from AEPA) was less satisfactory. Comparisons of the various estimates of the proportions of microbial-N in abomasal digesta suggested that the results obtained for protozoal-N by AEPA were overestimates. AEPA was found in mixed rumen bacteria which may have accounted in part for these overestimates. However, AEPA was not detected in any of the dietary ingredients.