In situ evaluation of the ruminal and intestinal degradability of extruded whole horse beans

Four rumen and proximal duodenum fistulated non-lactating Holstein cows were used to determine the effect of extrusion at 120 degrees C of whole horse beans (Vicia faba cv Talo) on in vitro nitrogen (N) solubility and in situ degradation of dry matter (DM) and crude protein (CP) in the rumen and intestine. Cows were fed a ration of 30% whole horse beans (WHB) and 70% Italian rye-grass hay. The degradation of DM and CP was estimated using nylon bags suspended in the rumen for 2, 4, 7, 16, 24 and 48 h; the effective ruminal degradability of DM and CP was evaluated assuming a ruminal outflow rate of 0.06/h. Bags incubated in the rumen for 16 h were introduced into the small intestine through the duodenal cannula and subsequently recovered in the feces. Extrusion of WHB reduced N-solubility in buffer solution (21.1 vs 74.9%). Processing diminished the effective rumen degradability of DM (74.6 vs 80.4%) and CP (70.2 vs 89.2%). Meanwhile, the amounts of DM and CP digested in the intestine increased: 9.6 vs 1.4% and 25.2 vs 3.0% respectively. Therefore, feeds containing extruded WHB increase the availability of dietary proteins in the intestine compared with diets containing raw WHB.     

A simplified management of the in situ evaluation of feedstuffs in ruminants: Application to the study of the digestive availability of protein and amino acids corrected for the ruminal microbial contamination

The ruminal effective degradability (RED) and intestinal effective digestibility (IED) for dry matter, crude protein (CP) and amino acids (AA) were estimated by a simplified in situ method using pooled samples from rumen-incubated residues, which represented the ruminal outflow of undegraded feed. The effect of microbial contamination in the rumen was corrected using ¹⁵N infusion techniques. Studies were carried out for soybean meal (SBM), barley grain (BG) and lucerne hay (LH) in three wethers cannulated in the rumen and the duodenum. Uncorrected values of RED for CP obtained either by mathematical integration or our simplified method were similar in all feeds. Microbial N in the pooled samples of SBM, BG and LH were 2%, 11% and 24% of total N, respectively. However, intestinal incubation eliminated this microbial charge by 100%, 99% and 88%, respectively. With microbial corrections, RED showed an increase, and IED showed a decrease, except for SBM. With this correction, intestinal digested CP was reduced by 2% in SBM, 13% in BG and 34% in LH. Corrected IED of AA was relatively similar in SBM (97–99%). However, large variations were observed in BG (74–93%) and in LH (10–88%). Digestion in the rumen and intestine changed the essential AA pattern. Overall, our results support that AA digestion is affected by the characteristics of their radicals and their contents in plant cell wall proteins. The accurate estimation of feed metabolisable AA or protein requires effective measures that are corrected by ruminal microbial contamination. The proposed in situ method largely simplifies these tasks and allows a more complete and less expensive feed evaluation.     

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 (k_p : 0.056/h) of the EC (ED_p) or additionally the rate of comminution and mixing (k_c : 0.130/h) of these particles (ED_cp). Ensiling increased ED_p (9.33%, p < 0.01) or ED_cp (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 k_p and k_c . 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.     

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).     

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 (k_c) and outflow. Results indicate that not considering k_c 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 k_c 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.     

Degradability characteristics of dry matter and crude protein of forages in ruminants

Twelve corn silages, 22 grass silages and 14 grass hays, obtained from various farms located in the lower Fraser Valley region of British Columbia, and 16 alfalfa hays, grown primarily in the Columbia basin of central Washington State, were evaluated using both the rumen and the mobile nylon bag in situ techniques. Nylon bags containing each forage were incubated in duplicate for 0, 2, 4, 8, 12, 24, 48, 72, or 96 h in two of six non-lactating Holstein cows fitted with rumen and duodenal cannulae. All forage types were evaluated in terms of the following dry matter (DM) and crude protein (CP) digestion characteristics: soluble fraction A, degradable fraction B, degradation rate, lag phase, and effective degradability. The mobile nylon bag technique was used to determine intestinal disappearance of DM and CP from the forages following pre-incubation in the rumen for 12 h. Significant (P < 0.05) differences in degradation characteristics occurred within all forages with regard to the soluble and potentially degradable DM and CP fractions. Soluble CP content in the rumen varied from 44.08 to 75.37% and from 18.74 to 65.38% in the corn and grass silages, respectively, and from 48.27 to 75.43% and from 30.13 to 65.95% in the alfalfa and grass hays, respectively. Significant differences within each forage type were also observed for the degradable CP in fraction B: 10.89 to 45.28% for corn silage, 20.72 to 82.77% for grass silage, 16.67 to 44.88% for grass hay and 25.44 to 62.93% for alfalfa hays. Significant differences (P > 0.05) were observed in fractional rates of ruminal DM degradation of the grass hays and corn silages. Significant differences did exist in the fractional rates of ruminal CP degradation within all forage types with the exception of alfalfa hays. Effective degradabilities of DM and CP were also significantly different between samples of a particular forage type. The mobile nylon bag data indicated that approximately 20% of the original CP in the grass silage, grass hay and alfalfa hay samples disappeared in the intestine and that there was significant variation between individual samples. On average, in the corn silage samples more than 10% of the original nitrogenous material disappeared in the intestine. The results presented in this study clearly demonstrate that the use of tabulated values for describing individual batches of forages in terms of their degradability characteristics is inaccurate since they may not reflect the particular forage being used in the ration and thus may lead to errors in diet formulation.     

Variation in ruminal in situ degradation of crude protein and starch from maize grains compared to in vitro gas production kinetics and physical and chemical characteristics

The objectives of this study were (1) to evaluate in situ ruminal dry matter (DM), crude protein (CP) and starch degradation characteristics and in vitro gas production (GP) kinetics using a set of 20 different maize grain genotypes and (2) to predict the effective degradation (ED) of CP and starch from chemical and physical characteristics alone or in combination with in vitro GP measurements. Maize grains were characterised by different chemical and physical characteristics. Ruminal in situ degradation was measured in three lactating Jersey cows. Ground grains (sieve size: 2 mm) were incubated in bags for 1, 2, 4, 8, 16, 24, 48 and 72 h. Bag residues were analysed for CP and starch content. Degradation kinetics was determined and the ED of DM, CP and starch calculated using a ruminal passage rate of 5%/h and 8%/h. The GP of the grains (sieve size: 1 mm) was recorded after 2, 4, 6, 8, 12, 24, 48 and 72 h incubation in buffered rumen fluid and fitted to an exponential equation to determine GP kinetics. Correlations and stepwise multiple linear regressions were evaluated for the prediction of ED calculated for a passage rate of 5%/h (ED5) for CP (EDCP5) and starch (EDST5). The in situ parameters and ED5 varied widely between genotypes with average values (±SD) of 64% ± 4.2, 62% ± 4.1 and 65% ± 5.2 for ED5 of DM, EDCP5 and EDST5 and were on average 10 percentage points lower for a passage rate of 8%/h. Degradation rates varied between 4.8%/h and 7.4%/h, 4.1%/h and 6.5%/h and 5.3%/h and 8.9%/h for DM, CP and starch, respectively. These rates were in the same range as GP rates (6.0–8.3%/h). The EDCP5 and EDST5 were related to CP concentration and could be evaluated in detail using CP fractions and specific amino acids. In vitro GP measurements and GP rates correlated well with EDCP5 and EDST5 and predicted EDCP5 and EDST5 in combination with the chemical characteristics of the samples. Equations can be used to obtain quick and cost effective information on ruminal degradation of CP and starch from maize grains.     

Effects of dietary crude protein and tannic acid on rumen fermentation,rumen microbiota and nutrient digestion in beef cattle

The objectives of the trial were to study the effects of dietary crude protein (CP) and tannic acid (TA) on rumen fermentation, microbiota and nutrient digestion in beef cattle. Eight growing beef cattle (live weight 350 ± 25 kg) were allocated in a 2 × 2 crossover design using two levels of dietary CP [111 g/kg dry matter (DM) and 136 g/kg DM] and two levels of TA (0 and 16.9 g/kg DM) as experimental treatments. Each experimental period lasted 19 d, consisting of 14-d adaptation and 5-d sampling. The impacts of dietary CP and TA on ruminal microbiota were analysed using high-throughput sequencing of 16S rRNA gene. Results indicated that no interactions between dietary CP and TA were found on rumen fermentation and nutrient digestibility. Increasing dietary CP level from 111 to 136 g/kg DM increased the ruminal concentrations of ammonia nitrogen (NH₃-N) (p < 0.01) and improved the CP digestibility (p < 0.001). Adding TA at 16.9 g/kg DM inhibited rumen fermentation and decreased the digestibility of dietary CP (p < 0.001), DM (p < 0.05) and organic matter (p < 0.01). Increasing the dietary CP level or adding TA did not affect the relative abundances of the major bacteria Firmicutes and Proteobacteria at the phylum level and Prevotella_1 and Christensenellaceae_R-7_group at the genus level, even though adding TA increased the Shannon index of the ruminal bacterial community. TA was partly hydrolysed to pyrogallol, gallic acid and resorcinol in rumen fluid and the inhibitory effects of TA on rumen fermentation and nutrient digestibility could have been resulted from the TA metabolites including pyrogallol, gallic acid and resorcinol as well as the protein-binding effect.     

Variation of lupin protein degradation in ruminants studied in situ and using chemical protein fractions

The main objective of this study was to evaluate the variability in in situ CP degradation characteristics of 15 batches lupin grains from nine genotypes in a standardised approach. This study also investigated whether differences in CP degradation can be described by protein fractionation using the Cornell Net Carbohydrate and Protein System (CNCPS) and also whether thermal processing of lupins has an effect on CP degradation in the rumen and analysed protein fractions. The rising political and consumer demand for milk products from dairy production systems based on domestic protein sources and the wide range of lupin types and varieties that can be chosen as protein feed in dairy nutrition requires research to determine the variability in CP degradation characteristics in the rumen. For CP degradation measurements, ground grains were incubated in the rumen of three lactating Jersey cows fitted with a ruminal cannula for different times from 2 to 48 h, and the washing loss of non-incubated samples was also measured. Protein fractions were analysed according to CNCPS and used for the estimation of ruminally degraded protein. In situ CP degradation parameters varied widely between untreated samples. The mean value for the washout fraction was 29.3% (from 16.4% to 43.6%). The potentially degradable fraction averaged 70.5% (from 55.6% to 83.7%), hence maximal degradation of CP was close to completeness. Mean degradation rate was 16.6%/h (from 12.6 to 21.0%/h). Variation in estimated parameters led to variation in the effective degradation (ED) averaging 76.6% (from 67.3% to 83.0%) when calculated assuming a ruminal outflow of 8%/h. Thermal treatment of lupins induced changes in degradation characteristics, primarily by lowering degradation rates, and also led to a significant reduction in ED. The ED calculated from analysed protein fractions averaged 10 percentage points higher than ED calculated from in situ parameters for untreated grains. The ED based on protein fractionation was also reduced by heat treatment, but the correlation with in situ based ED was poor. It can be concluded that the variation in ED indicates a potential to increase the amount of rumen undegraded protein without additional chemical or physical treatment and the effect of genetic factors and agronomic practices on ED of lupin grains should be investigated in systematic studies in the future.     

Effects of fibre concentration of diets consisting of hay and slowly degradable concentrate on ruminal fermentation and digesta particle size in mid-lactation dairy cows

Four multiparous ruminally cannulated Holstein cows (mean bodyweight [BW] 615 kg) in mid-lactation (103 days in milk and 32 kg milk x d(-1) at start of the experiment) were used in an one-factorial experiment to evaluate the effects of fibre level (19, 24, 28, 32 and 39% physically effective NDF [peNDF] in dry matter [DM]) in diets consisting of hay and slowly degradable concentrate on rumen fermentation patterns and digesta particle size, under a constant intake level (146 g DM x kg(-0.75). The different fibre concentrations in the diet were achieved by adjusting the hay to concentrate ratio. The above-mentioned levels of peNDF corresponded to 70, 60, 50, 40 and 25% concentrate in diet DM, respectively, and followed the lactation curve of the cows. The ruminal pH was positively and linearly correlated to the percentage of fibre (peNDF, NDF or CF) in ration DM with R2 of 0.76-0.88 (p < 0.001) for solid digesta (particle-associated rumen fluid, PARL), and R2 of 0.26-0.29 (p < 0.05) for fluid digesta (free rumen liquid, FRL). The lowest fibre level in the diet (19% peNDF) or the highest level of concentrate (70% on DM basis) caused pH values lower than 6.0 at almost all sampling times only in PARL but not in FRL, and significantly increased the proportion of large particles in rumen digesta, which in turn was reflected by a depression of fibre digestibility. A level of 24% peNDF or 60% concentrate in the diet maintained the ruminal pH higher than 6.0 and 5.8 in FRL and PARL, respectively. Therefore, the inclusion of more than 60% slowly degradable concentrate in dairy cows diets fed approximately 18 kg DM x d(-1) is discouraged. Based on the response of ruminal solid digesta to dietary fibre, it can be concluded that the recommendations of feeding a structural value > or =1 per kg DM (De Brabander et al. 1999) underestimated, and 400 g CF per 100 kg BW (Hoffmann 1990) overestimated the evaluation of structural effectiveness of the present diet.     

Effects of linseed lipids fed as rolled seeds, extruded seeds or oil on organic matter and crude protein digestion in cows

Effects of fatty acids of linseed in different forms, on ruminal fermentation and digestibility were studied in dry cows fitted with ruminal and duodenal cannulas. Four diets based on maize silage, lucerne hay and concentrates (65/10/25 dry matter (DM)) were compared in a 4 × 4 Latin square design experiment where the diets were: control diet (C), diet RL supplied 75 g/kg DM rolled linseeds, diet EL supplied 75 g/kg DM extruded linseeds, and diet LO supplied 26 g/kg DM linseed oil and 49 g/kg DM linseed meal. The diets did not differ in total organic matter (OM) and fibre digestibility, in forestomach and intestinal OM digestibility, and in duodenal N flow. Microbial N duodenal flow tended to be lower for RL versus C diet (P<0.1). Extrusion did not reduce ruminal crude protein (CP) degradation in vivo and in situ. Volatile fatty acid concentration and pattern, and protozoa concentration in the rumen, did not vary among diets. Results confirm the absence of a negative effect of a moderate supply of linseed on rumen function, as well as no effect of extrusion on its ruminal CP degradability.     

Digesta characteristics of dorsal, middle and ventral rumen of cows fed with different hay qualities and concentrate levels

The influence of fibre content of hay (H) and concentrate level (C) on local differences in the composition of ruminal digesta (ratio of solid to fluid digesta, DM, NDF, ADF and ADL content), particle size (MPL), specific gravity (SG) and fermentation (pH and concentrations of SCFA and bicarbonate) have been tested on two ruminally cannulated Friesian cows (520 kg BW) which were fed restricted, using individual cows as experimental units. Digesta samples were collected via cannula from three rumen layers: 5 to 10 cm (top) and 25-35 cm beneath the top of the particle mat (middle) and 5-10 cm above the rumen floor (bottom). For a main plot treatment (H x C), repeated samples were collected at four time intervals (1 h before and 2, 5 and 10 h after morning feeding) on each of two days. From top to bottom rumen the share of solid digesta mass (SM), DM and NDF contents of squeezed digesta fluid (SRF) and concentration of SCFA decreased (P < 0.05); pH and bicarbonate concentration increased (P < 0.05), while DM, NDF, ADF and ADL contents in SM, MPL and SG did not differ. Higher NDF content of hay (from 47-62%) increased SM, fibre fractions in SM, MPL, pH and concentration of bicarbonate in ruminal digesta, especially when 50% concentrate was given, while SG decreased. When the concentrate level was enhanced from 20 to 50%, digesta SM, MPL and the content of DM and NDF in SRF increased, while pH, concentrations of SCFA and acetate decreased when low-fibre hay was given. With longer time after feeding the digesta SM was reduced and fibre content in SM increased. The increase of the fibre content of hay reduced the possible negative effect of high concentrate level on the stratification of ruminal digesta. The decrease of the fibre content of hay promised better conditions for fibre digestion in the rumen when concentrate availability is limited.     

Ruminal degradability of 15N labelled ribonucleic acid in grass

The ruminal degradation of RNA in rye grass (Lolium perenne) was studied using the bag method. A non-lactating cow (BW 550 kg) fitted with a rumen cannula was used and fed twice daily at maintenance level with a chopped grass hay-based ration containing 30% ground barley. Rye grass, labelled during growth by fertilization with 15N2-urea (9.5 atom% 15N, 20 g N/m2), was cut at seven stages of growth and maturity and freeze-dried. RNA-N represented 6 to 17% of total N. Labelled grass samples (milled to 5.0 mm screen, 5.0+/-0.1 g DM) were incubated in polyester bags (100 x 200 mm, pore size 50 microm) in the rumen for periods of 1, 3, 6, 9, 12, 24, and 48 h. Data of N and RNA disappearances from the bags were fitted to an exponential equation to estimate parameters of degradation. The effective degradability of RNA in the rumen averaged 90+/-4%, for N it was 11% units lower (P < 0.001). Degradability of RNA was correlated to that of N (R2 = 0.92). Degradability of RNA (R2 = 0.96) and N (R2 = 0.93) decreased with increasing fibre content of grass. Increasing the fibre content by 1% diminished the degradability of RNA and N by 1.1% units and 2.4% units, respectively (P < 0.001). Assuming a microbial protein synthesis in the rumen of 150 g/kg DOM, a N: RNA ratio of 1:1.35 in rumen microbes and a rumen outflow rate of 0.06 h(-1), a model calculation indicates that about 9 to 19% of duodenal RNA are of dietary origin in animals fed grass. This should be taken into account for the calculation of microbial N on the basis of RNA as marker.     

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.     

Effects of the replacement of concentrate and fibre-rich hay by high-quality hay on chewing,rumination and nutrient digestibility in non-lactating Holstein cows

The aim of this study was to investigate the effects of high-quality hay with an elevated sugar content alone or with graded amounts of concentrate feed on chewing and ruminating activity, apparent total tract digestibility (ATTD) and ruminal pH at different time points after feeding in the free ruminal liquid (FRL) and the particle-associated ruminal liquid (PARL). Eight rumen cannulated non-lactating Holstein cows were arranged in a Latin square design in four experimental runs lasting 25 d each. The four diets tested were 60NQ (60% normal-quality hay + 40% concentrate), 60HQ (60% high-quality hay + 40% concentrate), 75HQ (75% high-quality hay + 25% concentrate) and 100HQ (100% high-quality hay). Normal and high-quality hays differed in sugar contents (11.3% vs. 18.7% in dry matter [DM]), neutral detergent fibre (NDF; 57.7% vs. 46.3% in DM), acid detergent fibre (ADF, 35.0% vs. 23.5% in DM) and crude protein (CP, 11.3% vs. 23.5% in DM). Data showed that ATTD of DM, CP, NDF and ADF was higher with the high-quality hay diets. Time spent eating was reduced with high-quality hay. However, time spent ruminating was longest in Group 100HQ. In all groups, ruminal pH of FRL and PARL decreased with time after the morning feeding. But 10 h later, pH of Group 100HQ was higher again compared with the other groups. Considering the average pH in FRL over all measured time points, cows in Groups 60NQ and 100HQ had higher pH values of 6.85 and 6.83, respectively. Regarding pH values in PARL, animals of Group 60NQ displayed the highest pH value (6.68), whereas the lowest value of 6.21 was found in Group 60HQ. Overall, results suggest that high-quality hay maintains the diet’s structural effectiveness by stimulating rumination and stabilising ruminal pH while greatly improving ATTD. However, the structural effectiveness of the high-quality hay gets impaired with increasing proportion of concentrate feed in the diet.     

Influence of condensed tannins from Brazilian semi-arid legumes on ruminal degradability,microbial colonization and ruminal enzymatic activity in Saanen goats

The present study aimed at determining the influence of condensed tannins present in the Brazilian legume species Mimosa hostilis, Mimosa caesalpinifolia and Bauhinia cheilantha on ruminal degradability, microbial colonization and enzymatic activity. Polyethylene glycol (PEG) was used to reduce the astringency and concentration of soluble condensed tannins. Four ruminally-cannulated Saanen goats (60 ± 8 kg BW) were fed, in two experimental periods, with a hay diet based on the studied legumes treated or non-treated with PEG. Voluntary intake, microbial colonization, DM, CP, NDF, and ruminal degradability of PEG treated and non-treated forage leaves, as well as pH, ammonia and 1,4 β-endoglucanase activity of the rumen content were evaluated. Astringency and soluble tannin concentration of the studied legumes were reduced by approximately 70% and 50%, respectively, with PEG treatment. Average DM intake was higher for the treated diet (16.76 g DM/kg BW/day against 13.06 g DM/kg BW/day). Percentile values for degradation parameters and for potential and effective degradabilities of DM, CP and NDF were also affected by the tannins, but at different intensities. Electron microscopic observations of ruminally-incubated legume leaves showed a more effective microbial colonization of PEG-treated leaves for all legume species. A decrease in pH and an increase in ammonia concentration and in endoglucanase activity in the ruminal content was also observed for PEG-treated diets at all sampling periods. Condensed tannins of the studied legume species have influenced the adhesion conditions, colonization and enzymatic activity of the microbial ecosystem, and consequently the ruminal degradation of the different dietary fractions. For this reason, the reduction in condensed tannin would be of great importance to improve the nutrition of ruminant feeding of these species.     

Effects of exogenous fibrolytic enzymes on in vitro ruminal fermentation of substrates with different forage:concentrate ratios

Batch cultures of mixed rumen micro-organisms were used to study the effects of three fibrolytic enzymes (xylanase from Trichoderma viride (XYL) and fibrolytic enzymes from Aspergillus niger (ASP) and Trichoderma longibrachiatum (TR)) on the fermentation of three substrates composed of grass hay:concentrate in the proportions (dry matter (DM) basis) of 0.7:0.3 (HF), 0.5:0.5 (MF) and 0.3:0.7 (LF). Enzymes were characterized for xylanase, endoglucanase, exoglucanase and amylase activities, and were supplied at rates of 40 and 80 enzymatic units/g substrate DM. In 8 h incubations, all enzymes increased (P=0.048 to P<0.001) the true degradability of substrate DM and the production of acetate, propionate, total volatile fatty acids (VFA) and gas. After 24 h incubation, some of the observed effects disappeared, but all enzymes still increased (P=0.028 to P<0.001) the degradability of substrate acid detergent fibre and the production of acetate, propionate and total VFA. For all enzymes, the effects on ruminal variables were less marked at 24 than at 8 h of incubation. Only few significant (P=0.044 to P=0.001) enzyme × substrate interactions were detected, although the magnitude of the response for each substrate varied with the enzyme. When considering the amount of organic matter apparently fermented (OMAF) and the methane:OMAF ratio as main variables, TR80 produced the greatest increase in OMAF (17.0%) for HF substrate, with ASP80 and TR40 having similar values (11.1 and 12.6%), and XYL and ASP40 showing no effects (P>0.05). A decrease (P<0.05) of methane:OMAF ratio was only found for TR80 at 8 h (17.4%). All enzymes, with the exception of ASP40, increased (P<0.05) OMAF at 8 h for MF substrate (11.3–25.4%), TR80 showing the greatest response. After 24 h of incubation, both doses of XYL and TR increased (P<0.05) OMAF (mean value 8.2%) and decreased methane:OMAF ratio (mean value 9.5%). All enzymes increased significantly OMAF with LF substrate at 8 h (7.5–19.9%), but after 24 h no effect (P>0.05) was detected on OMAF and methane:OMAF ratio. In general, few differences were detected between both doses of enzymes, which indicate than the used enzymes would be effective in enhancing ruminal degradation of substrates at a dose lower than 80 enzymatic units/g substrate DM.     

Effect of soybean oil availabilities on rumen biohydrogenation and duodenal flow of fatty acids in beef cattle fed a diet with crude glycerine

Soybean oil with different ruminal availability (whole soybeans (WS), soybean oil (SO) and calcium salts (CS)) was used to evaluate the fatty acid (FA) intake, rumen biohydrogenation (BH) and duodenal flow of FA in Nellore steers fed diets with crude glycerine (CG). Eight castrated Nellore steers were fitted with a ruminal and duodenal silicone cannula, and distributed in a double, simultaneous, Latin square 4 × 4 design with four diets and four experimental periods. Concentrates contained ground maize, urea, mineral salts, CG (100 g/kg DM) and soybean products with different availability of soybean oil: (1) no additional fat (CO), (2) WS, (3) SO or (4) CS. Fat supplementation was fixed to obtain 50 g ether extract/kg DM. Experimental treatments had no effect on DM intake, DM duodenal flow or ruminal turnover rate of C:16 FA. However, fat addition increased C:18 and turnover rates of total FA rumen (p < 0.05). CS resulted in lower C:18 turnover rates and lower ruminal BH of monounsaturated and unsaturated FA (UFA) than WS (p < 0.05). SO resulted in a greater duodenal flow of C18:0 (stearic acid), C18:1t-11 (vaccenic acid) and saturated FA than the WS and CS diets (p < 0.05). CS resulted in a higher duodenal flow of C18:3n-3 (linolenic acid) than WS (p < 0.05). The association of CG and calcium salts in Nellore steers was the best nutritional strategy to increase duodenal flow of healthier UFA, which may increase the deposition of these FA in meat. However, SO associated with CG association increased the duodenal flow of vaccenic acid, which is main precursor of endogenous synthesis of conjugated linoleic acids in tissues.     

Effects of isobutyrate on rumen fermentation, urinary excretion of purine derivatives and digestibility in steers

The objective of this study was to evaluate the effects of isobutyrate supplementations on rumen fermentation, urinary excretion of purine derivatives and feed digestibility in steers. Eight ruminally cannulated Simmental steers were used in a replicated 4 x 4 Latin square experiment. On DM basis, diet consisted of 60% corn stover and 40% concentrate. Dry matter intake (averaged 9 kg/d) was restricted to 90% of ad libitum intake. The four treatment groups received a daily dose of 0 (control), 8.4, 16.8 or 25.2 g isobutyrate per steer. With increasing isobutyrate supplementation total VFA concentration (range 64.2-74.0 mM) was significantly enhanced. The ratio of acetate to propionate (range 2.72-4.25) was also significantly increased due to the increase in actate production and decrease in propionate production. With increasing isobutyrate supplementation the ruminal degradation of NDF from corn stover was improved but the CP degradability of soybean meal was decreased. Furthermore, the isobutyrate supplementation caused a significantly increased urinary excretion of purine derivatives. Similarly, digestibilities of OM, NDF and CP in the total tract were significantly increased. The present results indicate that dietary supplementation with isobutyrate improved rumen fermentation and feed digestion in beef cattle in a dose-dependent manner. According to the conditions of this experiment, the optimum daily dose of isobutyrate was about 16.8 g/animal.     

Lotus corniculatus condensed tannins decrease in vivo populations of proteolytic bacteria and affect nitrogen metabolism in the rumen of sheep

Condensed tannins in forage legumes improve the nutrition of sheep by reducing ruminal degradation of plant protein and increasing crude protein flow to the intestine. However, the effects of condensed tannins in forage legumes on rumen bacterial populations in vivo are poorly understood. The aim of this study was to investigate the specific effects of condensed tannins from Lotus corniculatus on four proteolytic rumen bacteria in sheep during and after transition from a ryegrass (Lolium perenne)-white clover (Trifolium repens) diet (i.e., low condensed tannins) to a Lotus corniculatus diet (i.e., higher condensed tannins). The bacterial populations were quantified using a competitive polymerase chain reaction. Lotus corniculatus was fed with or without ruminal infusions of polyethylene glycol (PEG), which binds to and inactivates condensed tannins, enabling the effect of condensed tannins on bacterial populations to be examined. When sheep fed on ryegrass-white clover, populations of Clostridium proteoclasticum B316T, Butyrivibrio fibrisolvens C211a, Eubacterium sp. C12b, and Streptococcus bovis B315 were 1.5 x 10(8), 1.1 x 10(6), 4.6 x 10(8), and 7.1 x 10(6) mL(-1), respectively. When the diet was changed to Lotus corniculatus, the average populations (after 8-120 h) of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis decreased (P < 0.001) to 2.4 x 10(7), 1.1 x 10(5), 1.1 x 10(8), and 2.5 x 10(5) mL(-1), respectively. When PEG was infused into the rumen of sheep fed Lotus corniculatus, the populations of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis were higher (P < 0.01-0.001) than in sheep fed Lotus corniculatus without the PEG infusion, with average populations (after 8-120 h) of 4.9 x 10(7), 3.8 x 10(5), 1.9 x 10(8), and 1.0 x 10(6), respectively. Sheep fed the Lotus corniculatus diet had lower rumen proteinase activity, ammonia, and soluble nitrogen (P < 0.05-0.001) than sheep that were fed Lotus corniculatus plus PEG. The Lotus corniculatus diet reduced rumen nitrogen digestibility (P < 0.05) and ammonia pool size and increased the flow of undegraded feed nitrogen to the abomasum. The nitrogen intake, rumen non-ammonia nitrogen pool size, rumen microbial non-ammonia nitrogen pool size, and abomasal microbial non-ammonia nitrogen fluxes were similar both in sheep fed only Lotus corniculatus and in sheep fed Lotus corniculatus plus PEG, but nonmicrobial non-ammonia nitrogen flux to the abomasum was higher (P < 0.01) for the sheep fed only Lotus corniculatus. Although condensed tannins in Lotus corniculatus reduced the populations of some proteolytic bacteria, total ruminal microbial protein and microbial protein outflow to the abomasum were unchanged, suggesting a species-specific effect of condensed tannins on bacteria in the rumen.