Composition of free and adherent ruminal bacteria: inaccuracy of the microbial nutrient supply estimates obtained using free bacteria as reference samples and (15)N as the marker.

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.

In vitro efficiency of combined acid-heat treatments for protecting sunflower meal proteins against ruminal degradation.

Efficacy of combined acid-heat treatments to protect crude protein (CP) against ruminal degradation has not been extensively researched. Four in vitro trials (Daisy technology) with orthophosphoric and malic acids were performed to examine effects on protection of sunflower meal protein. In Trial 1, effects of the solution volume for adding two doses of orthophosphoric acid (0.4 and 1.2 eq/kg sunflower meal) were tested using five dilution volumes (80, 160, 240, 320 and 400 ml/kg of feed) for each acid dose. Samples were heated at 60°C. The quantity of CP that remained undegraded after 20 h in vitro (IVUCP) increased with the amount of acid added (P = 0.01). Increasing the dilution volume also tended (P = 0.065) to increase IVUCP. Therefore, a dilution volume of 400 ml/kg was employed in all further trials. In Trial 2, treatments with solutions of orthophosphoric and malic acids (1.2, 2.4, 3.6 and 4.8 eq/kg) and 60°C of drying temperature were used. Increased CP protection with increased acid doses was described. In this and further trials, higher protective effects of malic acid than orthophosphoric acid were also shown. In Trial 3, the effects of both these acids, four acid concentrations (0.6, 1.2, 1.8 and 2.4 eq/kg) and three levels of heat treatment required for drying the samples (100, 150 and 200°C for 60, 30 and 20 min, respectively) were evaluated. An interaction acid type × concentration × temperature was shown. In addition, interactions concentration × temperature was shown in each acid. With heat treatments of 100°C to 150°C, benefits were not obtained after increasing the acid dose over 0.8 eq/kg. The increase of the heat treatments to 200°C and the acid dose up to 1.2 eq/kg increased protection, but to exceed this dose did not improve protection. In Trial 4, available lysine, CP solubility in McDougall buffer and IVUCP were compared after treatment with water or solutions (0.8 eq/kg) of orthophosphoric or malic acids using 100°C and 150°C heat treatments as described in Trial 3. No effects on available lysine were observed. Both CP solubility and IVUCP were reduced to a greater degree by acids than by water treatment. The results showed a high effectiveness of acid-heat treatments. Levels of protection are dependent on the acid dose, its dilution, acid type and drying conditions.

Effects of feed intake on composition of sheep rumen contents and their microbial population size.

The present study was conducted to determine the effect of feed intake on the composition of the rumen contents of sheep and on their bacterial densities. Whole rumen contents were sampled after a period of continuous inter-rumen infusion of 15NH3 from four rumen-cannulated wethers successively fed on a hay-concentrate diet (2:1, w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight0.75. Total weight and chemical composition of rumen contents, as well as the distribution by size and chemical composition of particles, were determined. The populations of bacteria associated with the liquid (liquid-associated bacteria, LAB) and solid (solid-associated bacteria, SAB) fractions of rumen digesta and the distribution of SAB according to feed particle size were also examined. The greater feed intake caused an increase in the mass of the rumen contents, while its chemical composition did not change, except for a higher content of organic matter (P=0.023). The distribution of feed particles by size was similar at both levels of intake. The concentrations of neutral- and acid-detergent fibre in feed particles decreased and those of total, dietary, and microbial N increased, both with a quadratic response (P=0.001), as particle size decreased. The proportion of LAB in the microbial biomass of rumen digesta reached only 8.0 %. This proportion and the density of LAB were unaffected by the level of feed intake, whereas an apparent reduction (10.4 %) occurred with the SAB biomass in whole rumen contents. A systematic, but not significant, reduction (mean value 11.9 %) in the level of microbial colonisation in the different particle fractions with the increase of feed intake was also observed.

Effects of stage of harvest on the protein value of fresh lucerne for ruminants.

The ruminal degradation of dry matter (DM) and crude protein (CP) and the intestinal availability of CP of four fresh lucerne (Medicago sativa L.) samples, corresponding to a 3rd growing cycle and harvested at 2-week intervals, were determined. Rumen degradability, measured by the nylon bag technique, and rumen outflow rates were determined on three rumen-cannulated wethers. Intestinal digestibility was determined by the mobile bag technique on three duodenal fistulated wethers. Both groups of animals were fed a 2:1 lucerne hay to concentrate diet at an intake level of 40 g DM x kg(-1) BW0.75. The effective degradability (ED) of DM decreased with maturity in linear and quadratic form, as a consequence of a decrease in the soluble fraction and a similar increase in the undegradable materials. The resultant values were 0.795, 0.661, 0.600, and 0.576 for harvests at 2, 4, 6, and 8 weeks. The ED of CP showed the same trend. However, the variations (values of 0.896, 0.832, 0.791, and 0.817, respectively), were moderate and mainly due to the reduction of the proportion of soluble CP. The intestinal digestibility of CP of all samples showed a downward trend with the increase in the ruminal incubation time, as modelled according to a logistic function. The undegraded CP digested in the gut (Di) and therefore the effective intestinal digestibility (EID) were derived from this function according to the rumen outflow of undegraded CP. The effects of maturity on the mean values of Di, expressed as a proportion of the original CP content, were the opposite of those recorded for the ED of CP. These values were 0.067, 0.102, 0.115, and 0.089 for samples harvested at 2, 4, 6, and 8 weeks, respectively. Nevertheless, when Di was expressed as g CP x kg(-1) DM, these values (18.0, 17.4, 17.1, and 14.3, respectively) decreased in linear form. The same trend was observed for EID values, which represent 0.641, 0.609, 0.549, and 0.488, respectively. The change of the digestion site produced by the reduction of ED of CP was also associated with an increase in the undigested CP (values of 0.037, 0.066, 0.094, and 0.094, at the four harvesting times).

Composition of bacteria harvested from the liquid and solid fractions of the rumen of sheep as influenced by feed intake.

A study was conducted to determine the effect of the feed intake on the chemical composition of bacteria associated with the solid (solid-associated bacteria; SAB) and liquid (liquid-associated bacteria; LAB) fractions of rumen digesta, the digestive passage kinetics and their relationships. Whole rumen contents were sampled after a period of continuous infusion of 15NH3 from four ruminally-cannulated wethers provided successively with a hay-concentrate diet (2 : 1 w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight 0.75. SAB had a higher content of organic matter and total lipids (P < 0.001) and a similar N content as compared with LAB. The concentration of purines and 15N was lower (P = 0.011 and P < 0.001 respectively) in SAB than LAB, whereas the opposite was observed for the concentration of amino acids (mg/g DM; P = 0.031). An increase in feed intake produced an increase in the N (P = 0.034) and purine (P = 0.066) concentrations in bacteria and a decrease (P = 0.033) in their amino acid concentrations. Significant increases of rumen outflow rates of liquid and particles were also observed with increased feed intake. Rates of rumen outflow showed positive and negative linear relationships (P < 0.001) with the purine : N ratio and the proportion of amino acid on total N of bacteria respectively. SAB contained significantly higher proportions of leucine, isoleucine, lysine and phenylalanine and lower proportions of alanine, methionine and valine than LAB. The increase in feed intake also induced significant changes in the amino acid profile of bacteria, increasing arginine and methionine and decreasing alanine and glycine proportions. Results show that the outflow rate of rumen contents is a major factor in determining the proportion of nucleic acids and protein in rumen bacteria and explains some of the differences observed between LAB and SAB.

Estimation of intestinal digestibility of undegraded sunflower meal protein from nylon bag measurements. A mathematical model.

Ruminal nitrogen degradation and intestinal digestibility (ID) of the undegraded nitrogen of three sunflower meals were determined on three wethers fitted with rumen cannulae and T-type duodenal cannulae using nylon bags. Meals were obtained from semi-dehulled seeds by conventional hexane extraction (samples SD1 and SD2) or from whole seeds by a discontinuous procedure of pressing and hexane extraction (sample W), which causes a superior thermal effect. Therefore, effective degradability of nitrogen for the W sample (0.537) was lower (P < 0.001) than for conventional meals. Between the latter, SD2 had a lower value (P = 0.019) than SD1 (0.776 and 0.812, respectively). ID decreased in all meals (P < 0.001) as the ruminal incubation time (t) increased. This evolution could be described accurately by an exponential curve as ID = s + he(-kit). A method is proposed for estimating the proportion of undegraded ruminal nitrogen digested in the intestines (Di) from 1) the above equation, 2) the undegradable (r) and the insoluble and potentially degradable (b) nitrogen contents of the feed and the degradation rate of the last fraction (k(d)), and 3) the rumen outflow rate of particles (k(p)). The Di value is shown to be: [equation: see text] The percentages of nitrogen from digested feed in the intestines obtained with this method were 15.1, 17.2 and 39.0 for SD1, SD2 and W, respectively. Resulting effective ID values of undegraded nitrogen were 0.804, 0.767 and 0.844. Undigested nitrogen after ruminal and intestinal incubations decreased in linear and quadratic form in all meals as ruminal incubation time increased.