Previous studies have indicated that 15N enrichment of solid-associated bacteria (SAB) may be predicted from the same value in liquid-associated bacteria (LAB). The aims of this study were to confirm this and to measure the error in the nutrient supply from SAB, when LAB are used as the reference sample. For this purpose, the chemical and amino acid (AA) compositions of both the bacterial populations were studied in four experiments carried out on different groups of three rumen cannulated wethers. Diets (one in Experiments 1 and 4 and three in Experiments 2 and 3) had forage-to-concentrate ratios (dry matter (DM) basis) between 2 : 1 and 40 : 60, and were consumed at intake levels between 40 and 75 g DM/kg (BW)0.75. The bacteria samples were isolated after continuous infusion of (15NH4)2SO4 (40, 18, 30 and 25 mg 15N/day, in Experiments 1 to 4, respectively) for at least 14 days. In all experiments, SAB had consistently higher concentrations of organic matter (826 v. 716 g/kg DM, as average) and total lipids (192 v. 95 g/kg DM, as average) than LAB. Similar CP concentrations of both populations were observed, except a higher concentration in SAB than in LAB in Experiment 3. A consistent (in Experiment 4 only as tendency) higher AA-N/total N ratio (on average 17.5%) was observed in SAB than in LAB. The 15N enrichment in SAB was systematically lower than in LAB. On the basis of the results of all studies a close relationship was found between the 15N enrichment in SAB and LAB, which was shown irrespective of experiments. This relationship was established from Experiments 1 and 2 and the above cited previous results (n = 20; P < 0.001; R2 = 0.996), and then confirmed from the results of Experiments 3 and 4. These relationships between SAB and LAB demonstrate that CP supply from SAB is underevaluated by, on average, 21.2% when LAB are used as the reference. This underevaluation was higher for true protein and even higher for the lipid supply (32.5% and 59.6%, respectively, as an average of the four experiments). Large differences in AA profile were observed between SAB and LAB. The prediction equation obtained using 15N as the marker may be used to correct the errors associated with the traditional use of LAB as the reference sample, and therefore to obtain more accurate estimates of the microbial nutrient supply to the ruminants.

An in situ study was conducted on four rumen-cannulated wethers to determine (using 15N infusion techniques) the microbial contamination (mg bacterial DM or crude protein (CP)/100mg DM or CP) and the associated error on the effective degradability of fourteen feeds: barley and maize grains, soyabean and sunflower meals, full-fat soyabean, maize gluten feed, soyabean hulls, brewers dried grains, sugarbeet pulp, wheat bran, lucerne and vetch-oat hays, and barley and lentil straws. The DM or CP contamination in residues (M) fitted to single exponential or sigmoid curves. A general model (M=m (1−e−ft)j) was proposed to match this fit. Asymptotic values (m) varied from 2·84% to 13·3% and from 2·85% to 80·9% for DM and CP, respectively. Uncorrected results underestimated the effective degradability of both DM (P<0·05) and CP (P<0·01). For CP, this underestimation varied from 0·59% to 13·1%, with a higher but unascertainable error for barley straw. Excluding maize grain, the microbial contamination of both DM and CP, and the associated underestimation of the effective degradability of CP, were positively related to the cellulose content of the feed. The error in the effective degradability of CP was also negatively related to the CP content and its apparent effective degradability (R2 0·867). This equation allows easier and more accurate estimates of effective degradability, needed to improve protein-rationing systems.