J. M. Arroyo

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.

Effects of ensiling on in situ ruminal degradability and intestinal digestibility of corn forage

The effective degradability of dry matter (DM), crude protein (CP) and amino acids (AA), and the intestinal effective digestibility (IED) of DM and CP of a green forage corn (GC) and its silage (EC) were determined on freeze-dried samples using three rumen and duodenum cannulated wethers. Two rumen incubations with duplicate bags were performed for each feed. Rumen degradation was determined on one series of bags from each incubation. The other series was freeze-dried and used to determine IED using mobile nylon bags. Microbial contamination of rumen incubated residues (determined with 15N techniques) fitted exponential functions, which showed a greater microbial contribution in EC than in GC in the undegradable DM (18.6% vs. 13.5%) and CP (81.7% vs. 69.4%). Degradability was calculated considering the particle rumen outflow rate (kp : 0.056/h) of the EC (EDp) or additionally the rate of comminution and mixing (kc : 0.130/h) of these particles (EDcp). Ensiling increased EDp (9.33%, p < 0.01) or EDcp (5.30%, p = 0.062) of DM and was associated with losses of nitrogen and with large changes in the AA profile. It is necessary to correct the microbial contamination, because it represents 32.0% (GC) and 42.5% (EC) of the undegraded CP when using kp and kc . Ensiling caused higher degradabilities for some AA as well as large differences in the changes due to the rumen fermentation on the AA profile. However, it had only limited effects on the undegraded protein profile. Ensiling also reduced the IED of DM (23.3% vs. 14.6%; p = 0.057). In conclusion, results do not show losses of nutritive value by ensiling corn cut at vitreous grain stage.

Malic acid or orthophosphoric acid-heat treatments for protecting sunflower (Helianthus annuus) meal proteins against ruminal degradation and increasing intestinal amino acid supply.

The protection of sunflower meal (SFM) proteins by treatments with solutions of malic acid (1 M) or orthophosphoric acid (0.67 M) and heat was studied in a 3 × 3 Latin-square design using three diets and three rumen and duodenum cannulated wethers. Acid solutions were applied to SFM at a rate of 400 ml/kg under continuous mixing. Subsequently, treated meals were dried in an oven at 150°C for 6 h. Diets (ingested at 75 g/kg BW0.75) were isoproteic and included 40% Italian ryegrass hay and 60% concentrate. The ratio of untreated to treated SFM in the concentrate was 100 : 0 in the control diet and around 40 : 60 in diets including acid-treated meals. The use of acid-treated meals did not alter either ruminal fermentation or composition of rumen contents and led to moderate reductions of the rumen outflow rates of untreated SFM particles, whereas it did not affect their comminution and mixing rate. In situ effective estimates of by-pass (BP) and its intestinal effective digestibility (IED) of dry matter (DM), CP and amino acids (AAs) were obtained considering both rates and correcting the particle microbial contamination in the rumen using 15N infusion techniques. Estimates of BP and IED decreased applying microbial correction, but these variations were low in agreement with the small contamination level. Protective treatments increased on average the BP of DM (48.5%) and CP (267%), mainly decreasing both the soluble fraction and the degradation rate but also increasing the undegradable fraction, which was higher using orthophosphoric acid. Protective treatments increased the IED of DM (108%) and CP, but this increase was lower using orthophosphoric acid (11.8%) than malic acid (20.7%). Concentrations of AA were similar among all meals, except for a reduction in lysine concentrations using malic acid (16.3%) or orthophosphoric acid (20.5%). Protective treatments also increased on average the BP of all AA, as well as the IED of most of them. Evidence of higher increases for those AA showing a high resistance to degradation in the untreated meal were also observed. The total supply of metabolisable AA was increased by 3.87 times for sulphur-containing AA, whereas that of lysine was increased by 2.5 times, mainly because of lysine losses with heat treatments. These treatments and especially that with malic acid would be useful to increase the protein value of these meals but their combined use with lysine-rich protein concentrates would improve the metabolisable protein profile.

Sunflower meal and spring pea ruminal degradation protection using malic acid or orthophosphoric acid-heat treatments

The effects of solutions of malic or orthophosphoric acids (0.752 Eqg/kg of feed) and heat to protect proteins of sunflower meal (SFM) and spring pea (SP) against ruminal degradation were studied using particle transit, 15N infusion, in situ and electrophoretic techniques. Three wethers fitted with rumen and duodenum cannulae were successively fed three isoproteic diets including SFM and SP, untreated or treated with malic or orthophosphoric acids. Incubations of tested meals were only performed while feeding the respective diet. Estimates of the ruminally undegraded fraction (RU) and its intestinal digestibility of dry matter, organic matter (only for RU), crude protein and starch (only in SP) were obtained considering ruminal microbial contamination and particle comminution and outflow rates. When corrected for microbial contamination, estimates of RU and intestinal digestibility decreased in all tested fractions for both feeds. All RU estimates increased with the protective treatments, whereas intestinal digestibility-dry matter also increased in SFM. Low intestinal digestibility-crude protein values suggested the presence of antitrypsin factors in SP. Protective treatments of both feeds led to consistent increases in the intestinal digested fraction of dry matter and crude protein, being only numerically different for SP-starch (60.5% as average). However, treatments also reduced the organic matter fermentation, which may decrease ruminal microbial protein synthesis. Electrophoretic studies showed albumin disappearance in both SFM and SP, whereas changes in other RU proteins were more pronounced in SP than SFM.

Effects of the comminution rate and microbial contamination of particles in the rumen on in situ estimates of protein and amino acid digestion of expeller palm kernel and rapeseed meal

BACKGROUND Microbial corrected effective in situ estimates of ruminal undegraded fraction (RU) and intestinal effective digestibility (IED) of dry matter (DM), crude protein (CP) and amino acids (AA) of expeller palm kernel (EPK) and rapeseed meal (RSM) were measured on three rumen- and duodenum-cannulated wethers using ¹⁵N labelling techniques and considering ruminal rates of comminution (k(c)) and outflow (k(p)) of particles. RESULTS The lack of k(c) and microbial correction overestimated the RU of DM by 4.91% (EPK) and 9.88% (RSM). The lack of this correction also overestimated in both feeds the RU of CP, individual and total (TAA) AA as well as the IED of DM, CP, TAA and most AA. RU estimates were higher for CP than for TAA, but the opposite was observed for IED. The intestinal digested fraction was higher for CP than for TAA: 17.4% (EPK) and 13.8% (RSM). Digestion led to large changes in the essential AA profile in both feeds. CONCLUSION 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.

Protein value of cereals and cereal by-products for ruminants: a comparison between crude protein and protein-based estimates.

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 15N 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.

Ruminal degradation of cell wall associated nitrogenous compounds of several 15N-labelled feeds

BACKGROUND Ruminal in situ effective degradability (ED) of dry matter (DM), neutral (NDF) and acid (ADF) detergent fibres, total-N and NDF (NDIN) and ADF (ADIN) bound-N in sunflower seed (SS), wheat grain (WG) and wheat straw (WS) were measured in three ruminally cannulated sheep, correcting microbial N-contamination using the (15) N dilution technique modified to consider the (15) N supply to adherent bacteria. RESULTS The lack of correction for N-contamination under-evaluated ED estimates in 1.52% (total-N), 28.0% (NDIN) and 33.3% (ADIN) in SS and in 1.02% (total-N) and 4.43% (NDIN) in WG. In the remaining cases, this contamination prevented establishing apparent degradation kinetics and, therefore, errors were not measured. Microbial corrected ED estimates in SS were higher in total-N (0.917) than in NDIN (0.559) and ADIN (0.520), which showed similar values. This behaviour was also shown in WS (0.670, 0.386 and 0.426, respectively), whereas decreasing values were shown from total-N (0.917) to NDIN (0.830) and ADIN (0.482) in WG. CONCLUSION Results confirm that NDF and ADF procedures failed to remove large fractions of particle adherent microorganisms, under-evaluating the ED of NDIN and ADIN. Degradation of NDIN represented a significant part of the degraded N, whereas ADIN contribution was only negligible in WG.

Effects of the correction of particle microbial contamination and particle transit model in the rumen on in situ protein evaluation of grass hays

Effects of considering the particle comminution rate (kc) in addition to particle rumen outflow (kp) and the ruminalmicrobialcontaminationonestimatesofby-passandintestinaldigestibilityofDM,organicmatterandcrudeprotein were examined in perennial ryegrass and oat hays. By-pass kc-kp-based values of amino acids were also determined. This studywasperformedusingparticletransit,insituand 15 Ntechniquesonthreerumenandduodenum-cannulatedwethers.The above estimates were determined using composite samples from rumen-incubated residues representative of feed by-pass. Considering thecomminutionrate,kc,modifiedthecontribution of theincubatedresidues tothesesamples inbothhays and revealed a higher microbial contamination, consistently in oat hay and only as a tendency for crude protein in ryegrass hay. Notconsideringkcorrumenmicrobialcontaminationovervaluedby-passandintestinaldigestibilityinbothhays.Therefore, non-microbial-correctedkp-basedvaluesofintestinaldigestedcrudeproteinwereoverestimatedascomparedwithcorrected andkc-kp-basedvalues inryegrass hay(17.4vs4.40%) andinoathay(5.73vs0.19%).Both factors shouldbeconsideredto obtain accurate in situ estimates in grasses, as the protein value of grasses is very conditioned by the microbial synthesis derived from their ruminal fermentation. Consistent overvaluations of amino acid by-pass due to not correcting microbial contamination were detected in both hays, with large variable errors among amino acids. A similar degradation pattern of amino acids was recorded in both hays. Cysteine, methionine, leucine and valine were the most degradation-resistant amino acids.

Effects of correcting in situ ruminal microbial colonization of feed particles on the relationship between ruminally undegraded and intestinally digested crude protein in concentrate feeds

BACKGROUND In situ estimates of ruminally undegraded protein (RUP) and intestinally digested protein (IDP) of ten concentrates, uncorrected or corrected for the ruminal microbial colonization, were used to examine the effects of this correction on the relationship between IDP and RUP values. Both variables were established for three rumen and duodenum cannulated wethers using 15 N labeling-techniques and considering measured rates of ruminal particle comminution (kc ) and outflow (kp ). RESULTS A covariance analysis showed that the close relationship found between both variables (IDP = -0.0132 ± 0.00679 + 0.776 ± 0.0002 RUP; n = 60; P < 0.001; r = 0.960) is not affected by correcting for microbial colonization (P = 0.682). CONCLUSION The IDP content in concentrates and industrial by-products can be predicted from RUP values, thus avoiding the laborious and complex procedure of determining intestinal digestibility; however, a larger sample of feeds is necessary to achieve more accurate predictions. The lack of influence of the correction for microbial contamination on the prediction observed in the present study increases the data available for this prediction. However, only the use of corrected values may provide an accurate evaluation.

Amino acid availability in ruminants of cereals and cereal co‐products

BACKGROUND Microbial corrected in situ estimates of the ruminal undegraded fraction (RU) and intestinal effective digestibility (IED) of amino acids (AA), except tryptophan, of rye, wheat and corn grains, wheat bran, wheat and barley distilled dried grains 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 and outflow. RESULTS The lack of microbial correction led to overestimations of the intestinal digested fraction that rose with the increase in ruminal degradability. Thus these overestimations varied widely among feeds (from 4.3 to 32.1% for total analysed AA) and among AA. Digestion led to large changes in the AA supply that were greater in the rumen than in the intestine. The impact of these changes on the protein value is conditioned by the magnitude of the undegraded protein fraction. CONCLUSION Microbial contamination taking place in the rumen and changes in the AA supply with digestion should be considered to attain accurate estimates of AA digestion. Globally, digestion improved the AA supply in rye, wheat and wheat distilled dried grain and decreased it in corn and corn gluten feed by reducing the supply of valine and basic AA, especially lysine.