Previous studies have indicated that (15)N 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 ((15)NH(4))(2)SO(4) (40, 18, 30 and 25 mg (15)N/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 (15)N 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 (15)N 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; R(2) = 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 (15)N 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.

SUMMARYEffects of the correction of microbial contamination (using15N techniques) and of considering the comminution rate (kc) of particles in the rumen on effective estimates of the ruminally undegraded (RU) fraction and its intestinal effective digestibility (IED) were examined in a sample of dehydrated sugar beet pulp (DBP) generating composite samples (from rumen-incubated residues) representative of the chemical composition of RU. Tested fractions were dry matter (DM), organic matter (OM, tested only for RU), crude protein (CP) and amino acids (AA). The study was performed on three rumen and duodenum cannulated wethers fed with a 2 : 1 (fresh weight basis) chopped oat hay-to-concentrate diet supplied at 40 g DM/kg BW0·75in six equal meals per day. The DBP showed sigmoid degradation kinetics: the fractional degradation rate increased by 5·8 times as time (h) increased from 0 to∞. Thekcrate (measured in the diet concentrate) represented 5·74% of the total rumen retention time of particles. As a result, the RU of DM was over-evaluated by 6·53% whenkcwas not considered. Microbial contamination of RU was high as in DM as in CP. Therefore, the overestimation of RU of DM was increased to 12·2% when this contamination was not corrected. The lack of this correction also led to large over-evaluations of RU and IED of CP and AA. As a result, the overestimation of the intestinal digested fraction was 40·9% for CP and 45·0% for total analysed AA. This overestimation varied largely among AA (from 18·9 to 88·7%). Corrected proportions of RU and IED were also variable among AA. Hypotheses on the causes of this variability are given. Resultant changes in the AA profile of the intestinal digested protein had some negative impact on the supply of essential AA and cysteine without affecting lysine. This problem is limited because the microbial protein synthesized from DBP fermentation in the rumen is largely predominant in the AA supply to the host.

The lack of k(c) and microbial correction as well as CP-based results leads to considerable overestimations in the protein use of both feeds. Digestion aggravates the lysine deficiency of EPK but has global positive effects in the absorbed profile of RSM.

In situ estimates of ruminal undegraded fraction (RU) and effective intestinal digestibility (EID, corrected for microbial colonisation) of dry matter (DM), crude protein (CP) and total analysed amino acids (TAA) of rye, wheat and corn grains, wheat bran, wheat and barley distillers' dried grains with solubles (DDGS) and corn gluten feed were measured on three rumen and duodenum cannulated wethers using (15)N labelling techniques and considering ruminal rates of particle comminution (kc) and outflow. Results indicate that not considering kc and microbial colonisation led to considerable overestimations of RU which increased with feed ruminal degradation. Microbial colonisation may be also associated with overestimations of EID, whose estimates for DM, CP and TAA were predicted from parameters related with the ruminal escape of intestinally indigestible materials. The RU estimates were higher for TAA than for CP in grains, but the opposite was observed in by-products, whereas EID estimates were higher for TAA in all feeds. To obtain accurate protein values in these feedstuffs, it is required to consider both kc and ruminal microbial colonisation. The CP-based results underestimate the intestinally digested protein in grains and the opposite is evidenced in cereal by-products. Microbial protein synthesised in the rumen is largely the major fraction of the feedstuff protein value with the exception of DDGS.

Sodium sulphite is used in an optional way to remove insoluble proteins from neutral detergent fibre (NDF) residue. To determine whether the recovery of both NDF and insoluble nitrogen (N) in NDF solution (NDIN) are altered by its use, both parameters were measured in a set of 12 feeds, including cereal grains: maize (MG), rye (RG) and wheat (WG); cereal co-products: maize gluten feed (MGF), distilled dried grains from barley (DDGB) and wheat (DDGW) and wheat bran (WB); protein concentrates: rapeseed meal (RSM) and expeller palm kernel (EPK); dehydrated sugar beet pulp (DBP) and oat (OH) and ryegrass (RGH) hays. Associated effects on the in situ effective degradability (ED) of both NDF and NDIN were also studied in DDGW, WB, RSM, EPK, DBP, OH and RGH. Also, ED of acid detergent fibre (ADF) and its N (ADIN) were studied in hays. Errors due to microbial contamination in the rumen on the ED of NDF, ADF, NDIN and ADIN were also established in these last seven samples using 15 N infusion methods. Three rumen and duodenum cannulated wethers were used in the study. The sulphite use in NDF solution led to reductions (DDGB, DDGW, RSM and OH) and increases (RG, WG, WB and DBP) of the NDIN proportion, as well as the contribution of crude protein to NDF. These variations were associated with irregular effects on NDF residues and on ED of both NDIN and NDF. As a consequence, sulphite use does not assure the reduction of the insoluble protein contamination and it may even increase it. This methodology may also alter the degradability estimates of NDIN or NDF. Mean ruminal microbial contamination in NDF was 7·0, 10·8, 13·3, 5·4, 12·0, 35·3 and 20·0 g/kg in WB, DDGW, RSM, EPK, DBP, OH and RGH, respectively. The associated contents of microbial N in NDIN were: 59·3, 29·9, 26·2, 19·8, 37·3, 441 and 150 g/kg, respectively. Microbial contamination in ADF and ADIN (g/kg) was 3·6 and 94·5 in OH and 1·7 and 41·2 in RGH. Not correcting this contamination led to consistent undervaluations of ED of NDIN and NDF in all tested feeds, although errors only reached significance for NDIN in hays and DBP. Microbial-corrected ED of NDIN was 0·685, 0·826, 0·481, 0·389, 0·166, 0·718 and 0·425 in WB, DDGW, RSM, EPK, DBP, OH and RGH, respectively, whereas values for ADIN were 0·504 (OH) and 0·469 (RGH).