Meta-analysis of lactation performance in dairy cows receiving supplemental dietary methionine sources or postruminal infusion of methionine.

The objectives of our study were to evaluate the productive response to methionine supplementation in lactating dairy cows and to define a relationship between metabolizable Met (MP Met) intake and production. A database of 64 papers meeting the selection criteria was developed evaluating postruminally infused dl-methionine (9 papers with 18 control diets and 35 treatment comparisons), 2-hydroxy-4-methylthio butanoic acid (HMTBa) provided as either a liquid or Ca salt form (17 papers with 34 control diets and 46 treatment comparisons), Mepron (Evonik Industries, Essen, Germany; 18 papers with 35 control diets and 42 treatment comparisons), and Smartamine (Adisseo Inc., Antony, France; 20 papers with 30 control diets and 39 treatment comparisons). Dietary ingredients and their accompanying nutritional compositions as described in the reports were entered into the Cornell-Penn-Miner software to model the diets and to predict nutrients that were not reported in the original publication. Data were analyzed using a weighted analysis of response to supplementation compared with the intraexperiment control, as well as through a regression analysis to changing dietary MP Met. Data included in the analysis were from experiments published between 1970 and 2011 with cows supplemented with between 3.5 and 67.9 g of Met or its equivalent from HMTBa. Cows supplemented with Smartamine consumed more, whereas cows supplemented with Mepron consumed less DM compared with controls. Milk yield did not significantly respond to Met supplementation, although it tended to increase for cows supplemented with HMTBa and Mepron. Milk protein yield was increased due to supplementation from all sources or from infusion, and protein concentration was greater for all supplements or infusion of dl-Met, except for cows supplemented with HMTBa. Irrespective of Met source, milk protein yield increased 2.23 g of protein/g of MP Met until reaching the breakpoint. Milk fat yield was increased for Mepron and HMTBa, whereas milk fat concentration was increased for infused dl-Met and for cows supplemented with HMTBa. Based on regression analysis, response of milk fat yield to Met supplementation was not different for infused dl-Met, Mepron, and Smartamine (1.87 g of fat/g of MP Met), whereas the response to HMTBa was significantly greater at 5.38 g of fat/g of MP Met.

Short communication: salivary secretion during meals in lactating dairy cattle.

Four multiparous Holstein cows in midlactation were used in a 4 x 4 Latin square to evaluate whether source of forage influenced salivary secretion during eating in lactating dairy cows. The forages were allocated separately from the pelleted concentrates. Cows were offered 1 of 4 forages each period: barley silage, alfalfa silage, long-stemmed alfalfa hay, or chopped barley straw. Saliva secretion was measured during the morning meal by collecting masticates through the rumen cannula at the cardia of each cow. Rate of salivation (213 g/min) was not affected by forage source. However, the forage sources differed in eating rate (g of DM/min), which led to differences in ensalivation of forages (g of saliva/g of DM and g of saliva/g of NDF). On the basis of DM, ensalivation (g of saliva/g of DM) was greatest for straw (7.23) and similar for barley silage, alfalfa silage, and alfalfa hay (4.15, 3.40, and 4.34 g/g of DM, respectively). Higher ensalivation of straw could be accounted for by its higher neutral detergent fiber (NDF) content; ensalivation of NDF (g of saliva/g of NDF) was actually greatest for long-stemmed alfalfa hay (12.4) and similar for the other chopped forages (8.9). Cows consumed concentrate about 3 to 12 times faster than the various forages (DM basis), and ensalivation of concentrate was much lower (1.12 g of saliva/g of DM) than for forages. Feed characteristics such as particle size, DM, and NDF content affect salivary output during eating by affecting the eating rate. Slower eating rate and greater time spent eating may help prevent ruminal acidosis by increasing the total daily salivary secretion in dairy cows.

Effect of dietary Enterolobium cyclocarpum on microbial protein flow and nutrient digestibility in sheep maintained fauna-free, with total mixed fauna or with Entodinium caudatum monofauna.

Three groups of five wethers with ruminal and duodenal cannulas and maintained as either fauna-free (FF) or inoculated with total mixed fauna (TF) or Entodinium caudatum as a single-species monofauna (EN) were used in an experiment with two 28 d periods. In the first period, the sheep were fed a control barley-based diet (40:60 concentrate to silage DM) and in the second period the diet was supplemented with 187 g DM of Enterolobium cyclocarpum for the last 12 d of the period. The diets of period 1 and 2 were isonitrogenous. There was no effect of fauna on apparent ruminal and total tract organic matter and fibre digestion, but bacterial and microbial N flow and efficiency were improved in FF sheep compared to TF sheep. In period 2, protozoal numbers were reduced between 31 and 88 % 2 h after feeding E. cyclocarpum for the third to twelfth day of supplementation and by an average of 25 % in samples collected over the 24 h feeding cycle. Supplementation of the diet with E. cyclocarpum and the consequent protozoal reduction in TF and EN sheep improved the flow of non-ammonia N and bacterial N to the small intestine and the efficiency of microbial synthesis. However, E. cyclocarpum reduced ruminal organic matter digestion, especially in faunated sheep, and total tract organic matter, N and fibre digestion. Thus, a reduction in the protozoal cell numbers of 25 % was sufficient to achieve the beneficial effects of reduced fauna on the bacterial protein supply, but diet digestibility was reduced.

Duodenal flow and digestibility in fauna-free sheep and in sheep monofaunated with Entodinium caudatum or Polyplastron multivesiculatum.

Three groups of five rumen and duodenum cannulated fauna-free sheep were used in a 28 d experiment. One group remained fauna-free, whereas the second (EN) and third (PP) groups, respectively, were inoculated intraruminally with the protozoan species Entodinium caudatum and Polyplastron multivesiculatum. Rumen fluid, duodenal digesta and faecal samples were collected during the last 12 d. The flow of digesta to the duodenum was determined using Yb and Co as dual-phase markers. (15)Nitrogen and phosphatidylcholine were used as markers to calculate the duodenal flow of bacterial and protozoal N, respectively. Results showed an increase (P < 0.1) in the rumen concentration of NH3-N and total volatile fatty acids, and a decrease (P < 0.05) in the duodenal flow of non-NH3-N and bacterial N in sheep with EN and PP monofaunas, compared with fauna-free sheep. There were no differences (P > 0.05) in these variables between the two monofauna groups. Protozoal N accounted for 8 % of the duodenal non-NH3-N flow in the EN-monofaunated sheep, whereas no such flow was detected in the PP-monofaunated sheep. Apparent rumen digestibility of organic matter, neutral detergent fibre and acid detergent fibre were similar (P > 0.05) in the monofaunated groups of sheep, but rumen acid detergent fibre digestibility was higher (P < 0.05) in the monofaunated than in the fauna-free groups. Experimental results suggested that, unlike EN, the PP monofauna might not contribute to the duodenal flow of microbial protein, whereas both monofaunas showed a virtually equal degree of predation on rumen bacteria.

Effect of grain processing and silage on microbial protein synthesis and nutrient digestibility in beef cattle fed barley-based diets.

Effects of the extent of grain processing and the percentage of silage in barley-based feedlot diets on microbial protein synthesis and nutrient digestibility were evaluated using four steers (initial BW of 442 +/- 15 kg) with ruminal and duodenal cannulas. The experiment was a 4 x 4 Latin square with four periods of 21 d each. Dietary treatments were arranged as a 2 x 2 factorial with two levels of barley silage (20 and 5% DM basis) and two degrees of barley grain processing (coarsely and flatly steamrolled to a processing index [PI] of 86 and 61%, respectively). The PI was quantified as the volume weight of the barley grain after processing, expressed as a percentage of the volume weight prior to processing. Digest a flow (Yb) and microbial (15N) markers were continuously infused into the rumen for a period of 13 d. Ruminal, duodenal, and fecal samples were collected at various times over the last 6 d of marker infusion. Diurnal ruminal pH was measured for 48 h. Intake of DM averaged 1.8% of BW, and was not different among the dietary treatments (P > 0.10). Ruminal starch digestibility was higher (P < 0.05) for the more extensively processed grain and tended (P < 0.10) to be highest when the more extensively processed grain was combined with 5% barley silage. In contrast, ruminal fiber digestibility for the 5% silage diets was reduced (P < 0.05) when the grain was more extensively processed. There was, however, no effect of grain processing on ruminal OM digestibility (P > 0.10), and hence, no inhibitory effect on microbial N flow to the intestine (P > 0.10). There was also no effect of the level of silage on microbial N flow (P > 0.10), but there was a tendency for improved efficiency of microbial protein synthesis for the 20% silage diets (P = 0.072). Ruminal escape of nonmicrobial N (P = 0.003) was greater, and thus, protein flow to the intestine was greater for the 5% silage diets. Diurnal ruminal pH was lower (P < 0.05) for 11 of the 24 hourly time points in steers fed the 5% silage diets than those fed the 20% silage diets. In conclusion, barley grain rolled to a PI of 86 to 61% and combined with 20 and 5% barley silage had little effect on microbial protein supply. Microbial protein supply was not inhibited when the barley grain was extensively processed (PI of 61%) and the silage was limited to only 5% of the diet DM, but feed intake of steers in this study was lower than would be expected in the feedlot.

Effects of particle size of alfalfa-based dairy cow diets on chewing activity, ruminal fermentation, and milk production.

Effects offorage particle size measured as physically effective NDF and ratio of alfalfa silage to alfalfa hay of diets on feed intake, chewing activity, particle size reduction, salivary secretion, ruminal fermentation, and milk production of dairy cows were evaluated using a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. The diets consisted of 60% barley-based concentrate and 40% forage, comprised either of 50:50 or 25:75 of alfalfa silage:alfalfa hay, and alfalfa hay was either chopped or ground. Various methods were used to determine physically effective NDF content of the diets. Cows surgically fitted with ruminal and duodenal cannulas were offered ad libitum access to these total mixed diets. The physically effective NDF content of the diets was significantly lower when measured using the Penn State Particle Separator than when measured based on particles retained on 1.18-mm screen. Intake of DM was increased by increasing the ratio of silage to hay but was not affected by physically effective NDF content of diets. Eating time (hours per day) was not affected by the physically effective NDF content of diets, although cows spent more time eating per unit of DM or NDF when consuming high versus low alfalfa hay diets. Ruminating time (hours per day) was increased with increased physically effective NDF content of the diets. Rumen pH was affected more by changing dietary particle size than altering the ratio of silage to hay. Feeding chopped hay instead of ground hay improved ruminal pH status: time during which ruminal pH was above 6.2 increased and time during which ruminal pH was below 5.8 decreased. Milk production was increased by feeding higher concentrations of alfalfa silage due to increased DM intake, but was not affected by dietary particle size. Feed particle size, expressed as mean particle length or physically effective NDF was moderately correlated with ruminating time but not with eating time. Although physically effective NDF and chewing time were not correlated to mean rumen pH, they were negatively correlated to the area between the curve and pH 5.8, indicating a positive effect on reducing the risk of acidosis. Milk fat content was correlated to rumen pH but not to physically effective NDF or chewing activity. These results indicate that increasing physically effective NDF content of the diets increased chewing activity and improved rumen pH status but had limited effect on milk production and milk fat content.

The effect of fibrolytic enzymes sprayed onto forages and fed in a total mixed ratio to lactating dairy cows.

We investigated the effect of spraying different combinations of fibrolytic enzymes onto forages on their nutritive value for lactating cows. Holstein cows were fed a TMR consisting of 30% corn silage, 15% alfalfa hay, and 55% concentrate (dry matter basis). During a 12-wk treatment period, the forages were treated with no enzymes (control), cellulase D and sultanas B, or cellulase D and xylanase C. Enzymes were diluted in water and sprayed onto the forages while mixing. Both combinations of enzymes supplied similar amounts of fibrolytic activity based on classical enzyme assays conducted at 50 degrees C. Cows fed forages treated with cellulase D and xylanase B tended to produce more 3.5% FCM (+2.5 kg/d) than did cows fed the untreated forages. Dry matter intake, milk production, milk fat, and milk protein were unaffected by treatment. In vitro production of gas from forages treated with enzymes was greater than from untreated forage, but 96-h volatile fatty acid production was not different among treatments. With an alternative enzyme assay based on the depolymerization of dyed substrate at 40 degrees C, activity of xylanase C was greatest at a pH of 6.5 but was substantially reduced as the pH of the assay was decreased. In contrast, xylanase B showed highest activity at pH 5 and enzyme activity was twice that of xylanase B at pH 5.5 and 6. Overall, the results of this study provide more evidence that fibrolytic enzymes can be used to improve milk production in lactating cows.

Effects of particle size of alfalfa-based dairy cow diets on site and extent of digestion.

Effects of ratio of alfalfa silage to alfalfa hay and forage particle size on nutrient intakes, site of digestion, rumen pools, and passage rate of ruminal contents were evaluated in a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. The diets consisted of 60% barley-based concentrate and 40% forage made up either of 50:50 or 25:75 of alfalfa silage:alfalfa hay and alfalfa hay was either chopped or ground. Lactating dairy cows surgically fitted with ruminal and duodenal cannulas were used and offered ad libitum access to a total mixed ration. Intakes of nutrients were increased by increasing ratio of silage to hay but were not affected by particle size of forage. Change in ratio of silage to hay of diets did not affect site and extent of digestion. However, increased forage particle size of the diets improved digestibility of fiber and N in the total tract, and as well as digestibility of organic matter, starch, and acid detergent fiber in the intestine. There was a shift of starch digestion from the rumen to the intestine when forage particle size was increased, although total digestion of starch was not changed. Ruminal microbial protein synthesis and microbial efficiency also improved with increasing forage particle size. Cows fed ground hay versus chopped hay had significantly lower rumen wet mass regardless of the ratio of silage to hay. Reduced forage particle size also lowered ruminal nutrient pool size for cows fed the high silage diet. Ruminal passage rates of liquid and solid were decreased by reducing the ratio of silage to hay, and retention time of solids in the total tract was shortened by reducing forage particle size. These results indicate that manipulating ratio of silage to hay in the diets of dairy cows changed feed intake but had little effect on digestion. In contrast, increased forage particle size in dairy cow diets improved fiber digestion and microbial protein synthesis in the rumen, and shifted starch digestion from the rumen to the intestine. Dietary particle size, expressed as physically effective neutral detergent fiber, was a reliable indication of ruminal microbial protein synthesis and nutrient digestion.

Evidence of quorum sensing in the rumen ecosystem: detection of N-acyl homoserine lactone autoinducers in ruminal contents.

Acyl-homoserine lactone (AHL) based quorum-sensing systems are widespread among gram-negative bacteria, particularly in association with plants and animals. As yet, there have been no reports of AHL signaling in the anaerobic rumen environment, an ecosystem of great complexity in which cell-cell signaling is likely to occur. We detected multiple AHL autoinducers in the rumen contents of 6 out of 8 cattle fed a representative selection of diets. The signals were not associated with feed. Surprisingly, no pure cultures produced AHLs in vitro when grown under the laboratory conditions we tested. Our observations suggest that either (a) a factor specific to the rumen ecosystem is required for the rumen isolates we tested to produce AHLs or (b) a strain (or strains) that we were not able to culture but which grows to a high cell density in the rumen produces the AHLs we detected.

Rumen degradation and availability of various amounts of liquid methionine hydroxy analog in lactating dairy cows.

Ruminal escape of various amounts of methionine hydroxy analog [D,L-2-hydroxy-4-(methylthio)-butanoic acid (HMB)] was measured in an experiment designed as a 4 x 4 Latin square using four lactating dairy cows with cannula in the rumen and duodenum. The cows were fed a diet composed of corn silage, alfalfa haylage, rolled barley grain, canola meal, and blood meal, three times per day. The cows were fed the liquid analog each day for 1 wk before the experiment was started. On the day of the experiment, each cow received an intraruminal bolus dose of 0, 25, or 50 g of the liquid analog (Alimet feed supplement, 88% HMB) or 51.2 g of a dry calcium salt of the analog (86% HMB; MHA) mixed with 0.5 kg of ground barley grain. A liquid phase marker (Co-EDTA) was administered as a bolus dose into the rumen at the time of administration of the methionine hydroxy analogs. Rumen and duodenal contents, and blood serum were collected at 0, 1, 3, 6, 9, 12, and 24 h relative to the time of dosing. Rumen and duodenal samples were analyzed for Co and HMB, and serum was analyzed for free methionine. Fractional rate constants for the passage of the liquid marker (k(p)) and the decline of HMB concentration in the rumen (k(rHMB)) were determined by nonlinear regression. Liquid passage from the rumen was similar among the four analog treatments (0.136 +/- 0.012/h; mean +/- SEM). Ruminal escape of HMB as a percentage of the dose (100% x k(p)/k(rHMB)) did not differ between cows receiving 25, 50, and 51.2 g of the methionine analogs (42.5, 41.0, and 34.9 +/- 9.0%, respectively) and averaged 39.5%. Duodenal appearance of HMB as a percentage also did not differ between cows receiving 25, 50, and 51.2 g of the methionine analogs (16.2, 26.8, and 22.7%, respectively) and averaged 22%. Omasal absorption of HMB was variable ranging from 12.3 to 26.3% and averaged 17.6%. Serum methionine concentration peaked at 3 and 6 h after dosing and increased in proportion to the amount of the analog administered. It was concluded that 39.5% of the methionine hydroxy analog escaped rumen degradation, the percentage of the dose that escaped the rumen was not affected by the amount or form of the methionine analog fed, and the analog that escaped ruminal degradation was likely absorbed and metabolized to methionine.

Effect of a fibrolytic enzyme preparation from Trichoderma longibrachiatum on the rumen microbial population of dairy cows.

The effects of supplementing a dairy cow diet with incremental levels of a fibrolytic enzyme preparation (preparation B) from Trichoderma longibrachiatum on the rumen microbial population were investigated. Two cows fitted with rumen cannulae were each fed a diet containing barley-based concentrate (52%), maize silage (29%), and chopped alfalfa hay (19%), supplemented with 0, 1, 2, 5, or 10 L of preparation B per tonne of dry matter (DM). Preparation B stimulated numbers of total viable bacteria in a quadratic manner (P < 0.05), to approximately 230, 330, 390, and 250% at 1, 2, 5, and 10 L x t(-1) DM, respectively. Preparation B increased the numbers of cellobiose-utilizing (P < 0.01), xylanolytic (P < 0.05), and amylolytic bacteria (P < 0.05), but had no effect (P > 0.05) on numbers of cellulolytic bacteria. However, when bacterial numbers enumerated on each substrate were expressed as a proportion of total viable bacterial numbers, only cellobiose utilizers were stimulated, and this stimulation was limited to the 1 L x t(-1) DM level of preparation B (P < 0.05). The results of this study demonstrate that the inclusion of an exogenous fibrolytic enzyme preparation in dairy cow diets increased the numbers of rumen bacteria that utilize hemicelluloses and secondary products of cellulose digestion.

Effects of grain processing, forage to concentrate ratio, and forage particle size on rumen pH and digestion by dairy cows.

Dietary factors that alter the intake of effective fiber were evaluated for their effects on rumen fermentation, digestion, and milk production using a double 4 x 4 quasi-Latin square design with a 2(3) factorial arrangement of treatments. The dietary factors were extent of barley grain processing, coarse (1.60 mm) or flat (1.36 mm); forage-to-concentrate (F:C) ratio, low (35:65) or high (55:45) (dry matter basis); and forage particle length, long (7.59 mm) or short (6.08 mm). Eight lactating cows with ruminal and duodenal cannulas were offered ad libitum access to a total mixed diet and milked twice daily. Dry matter intake was increased by increasing the extent of grain processing. Mean rumen pH was lower for cows fed flatly rolled barley than for cows fed coarsely rolled barley, whereas F:C ratio or forage particle size had no effect on rumen pH. Rumen pH was not correlated with effective NDF intake but tended to be correlated with digestibility of starch in the rumen. Total tract digestibilities of dry matter, organic matter, starch, and neutral detergent fiber were increased by feeding flatly rolled barley or low F:C ratio diets. Milk yield and milk protein content were higher in cows fed flatly rolled barley or low F:C ratio diets. Milk fat content tended to increase with high F:C ratio or long forage particle length but was reduced by feeding flatly rolled barley. In this study, extent of grain processing and intake of ruminal available starch were the most influential factors affecting milk production. Reducing the ratio of F:C improved total digestion and actual milk production. Forage particle length had minimal impact on digestibility and milk production.

Effect of dietary factors on distribution and chemical composition of liquid- or solid-associated bacterial populations in the rumen of dairy cows.

Effects of dietary factors including kernel thickness of processed barley grain, ratio of forage to concentrate, and forage particle length on chemical composition of bacteria, bacterial colonization of feed particles and distribution in the rumen, and duodenal flow of bacteria in dairy cows were evaluated. The experiment was designed as a double 4 x 4 quasi-Latin square with a 2 x 2 x 2 factorial arrangement of treatments using eight lactating cows with ruminal and duodenal cannulas. Barley grain was steam-rolled to two thicknesses: coarse (1.60 mm) or flat (1.36 mm); ratio of forage:concentrate was low (35:65) or high (55:45) (DM basis); and forage particle length was long (7.59 mm) or short (6.08 mm). Cows were offered ad libitum access to a total mixed diet. Chemical composition was different (P < 0.01) between liquid-associated bacteria (LAB) and solid-associated bacteria (SAB). Reduced barley thickness increased (P < 0.05) N content and 15N enrichment, but a high ratio of forage:concentrate decreased (P < 0.01) 15N enrichment of both the LAB and SAB. Significant differences between AA composition of the LAB and SAB were observed for 15 out of 17 AA studied. Bacterial colonization was linearly increased (P < 0.01) from about 5 to 70% as particle length of rumen contents was reduced from 3.35 mm to 0.15 mm (sieve size). The degree of colonization on each fraction of the rumen particulate matter was only affected (P < 0.10) by the ratio of forage:concentrate, with consistently higher (P < 0.10) bacterial colonization noted for high than for low forage:concentrate diets. Of the total bacterial mass within the rumen, less than 20% was associated with the liquid and over 70% was associated with the small particles that passed through the 0.6-mm sieve. Although the bacterial pool in the rumen was lower (P < 0.04) when flatly rolled barley rather than coarsely rolled barley was fed, bacterial flow to the duodenum was greater (P < 0.10) with increasing ratio offorage:concentrate. The present results confirm the differences of chemical composition and biomass for LAB and SAB. Manipulation of dietary factors such as ratio of forage:concentrate have the potential to alter bacterial colonization of rumen particles and the relative proportion of LAB to SAB, which were positively correlated to bacterial flow to the duodenum.

Effects of barley grain processing on the site and extent of digestion of beef feedlot finishing diets.

Effects of extent of barley rolling on chewing activities, ruminal fermentation, and site and extent of digestion were evaluated for feedlot finishing cattle diets in a 4 x 4 Latin square design. Four Jersey steers (452 kg), cannulated in the rumen and duodenum, were used. Barley grain was temper-rolled to four extents: coarse, medium, medium-flat, and flat, which were expressed as processing index (PI, volume weight of barley after processing expressed as a percentage of its volume weight before processing, DM basis) and equivalent to 82, 75, 70, and 65%, respectively. Diets consisted of 9.7% barley silage, 86% temper-rolled barley, and 4.3% other ingredients (DM basis). Steers were offered ad libitum access to a total mixed ration once daily. Dry matter intake was not affected (P > 0.15) by PI of barley. Digestibility of OM in the rumen and in the total tract were numerically lower (P = 0.13) for steers fed coarsely rolled barley than for steers fed more extensively processed barley. Digestibility of starch in the total tract was linearly increased (P = 0.02) with grain processing, but NDF digestion was not affected by processing (P > 0.15). Digestibility of CP did not differ in the rumen but tended (P = 0.08) to increase in the total tract with increased processing of barley. Flow of microbial nitrogen to the duodenum was approximately one-third lower (linear effect, P = 0.06) for steers fed coarsely rolled barley than for steers fed further rolled barley. Increased grain processing tended to decrease (linear effect, P = 0.08) rumination time without affecting eating time. These results indicate that optimal degree of rolling for barley fed to feedlot cattle corresponded to a PI of 75% or lower. Coarsely rolled barley is not recommended because it resulted in the lowest digestibility and lowest microbial protein synthesis. Processing barley to attain a PI less than 75% resulted in marginal improvements in feed digestion, but rumination time decreased, which could lead to problems associated with acidosis if lower-fiber diets are used.

Resistance of feed enzymes to proteolytic inactivation by rumen microorganisms and gastrointestinal proteases.

Potential feed enzyme additives for ruminants were tested in vitro for their stability to ruminal microbial and gastrointestinal proteolysis. Four commercial preparations from Trichoderma longibrachiatum (A, B, C, and D) and one from an undisclosed source (E) were incubated up to 6 h with ruminal fluid taken from four lactating dairy cows before or 2 h after feeding. The stability of preparation B was also tested in the presence of pepsin at pH 3 and pancreatin at pH 7. Cellulase (EC 3.2.1.4), cellulose 1,4-beta-cellobiosidase (EC 3.2.1.91), beta-glucanase (EC 3.2.1.6), xylanase (EC 3.2.1.8), beta-glucosidase (EC 3.2.1.21), and beta-xylosidase (EC 3.2.1.37) activities were monitored throughout the incubations. Polysaccharidase activities of all enzyme preparations were remarkably stable in ruminal fluid taken after feeding. Ruminal fluid obtained before feeding inactivated the polysaccharidases in preparations B and D to a greater extent than ruminal fluid obtained after feeding. Cellulase and cellulose 1,4-beta-cellobiosidase activities were the least stable, declining (P < 0.05) by 35 and 60% for preparations B and D, respectively. Xylanase activity of preparation D decreased (P < 0.05) by up to 30% after 6 h of incubation, whereas beta-glucanase activity was not affected. The ability to degrade exogenous enzymes also differed among cows (P < 0.05). Pepsin and acid (pH 3.0) did not affect polysaccharidases in preparation B but decreased glycosidase activities by 10 to 15% (P < 0.05) after 1 h of incubation. Pancreatin, at the maximum concentration used, inactivated cellulase, cellulose 1,4-beta-cellobiosidase, and xylanase activities at a rate of 0.55, 1, and 0.45%/min, respectively. beta-Glucosidase and beta-xylosidase activities decreased by 1 and 0.75%/min, respectively. Partial proteolysis of cellulase, cellulose 1,4-beta-cellobiosidase, and xylanase by pancreatin produced a transient increase in activity. This twofold increase for cellulase and fourfold increase for cellulose 1,4-beta-cellobiosidase was directly proportional to pancreatin concentration. These results suggest that the enzyme feed additives tested were stable in the rumen of animals after feeding. Exogenous enzymes are likely to be more susceptible to the host gastrointestinal proteases in the abomasum and intestines than to ruminal proteases. However, exogenous polysaccharidases may survive for a considerable period of time in the small intestine and they probably maintain activity against target substrates in this environment.

Ruminal degradability, intestinal disappearance, and plasma methionine response of rumen-protected methionine in dairy cows.

Bioavailability of Met from a rumen-protected Met product was evaluated in two experiments using three ruminally and duodenally cannulated lactating (experiment 1) and nonlactating (experiment 2) dairy cows. In the first experiment, the ruminal in situ and mobile bag technique was used to assess ruminal degradability and intestinal disappearance of Met from the protected Met product. Effective ruminal degradability of Met at a ruminal outflow rate of 0.11/h was 21.7%. Combining effective ruminal degradability with intestinal digestibility yielded an estimate of Met availability of 25%. In the second experiment, designed as a 3 x 3 Latin square, Met availability was assessed by determining the response of plasma Met to supplementation of the protected Met product relative to that of duodenally administered Met. The periods were 1 wk with cows fed a meal containing 0, 20, or 63 g of protected Met on d 1 and infused intraduodenally with 10.7 g of Met on d 4. Blood was collected at various times relative to the time of oral dosing and the commencement of the duodenal infusion. Plasma Met response measured as area under the curve increased linearly with increasing protected Met. The response of plasma Met increased by 33 and 65.5% of the control values for 20 and 63 g of protected Met, respectively. Intestinal bioavailability of Met in the protected Met product ranged from 22 to 34%.

Effects of an exogenous enzyme preparation on microbial protein synthesis, enzyme activity and attachment to feed in the Rumen Simulation Technique (Rusitec).

The effects of an exogenous enzyme preparation, the application method and feed type on ruminal fermentation and microbial protein synthesis were investigated using the rumen simulation technique (Rusitec). Steam-rolled barley grain and chopped alfalfa hay were sprayed with water (control, C), an enzyme preparation with a predominant xylanase activity (EF), or autoclaved enzyme (AEF) 24 h prior to feeding, or the enzyme was supplied in the buffer infused into the Rusitec (EI). Microbial N incorporation was measured using (15NH4)2SO4 in the buffer. Spent feed bags were pummelled mechanically in buffer to segregate the feed particle-associated (FPA) and feed particle-bound (FPB) bacterial fractions. Enzymes applied to feed reduced neutral-detergent fibre content, and increased the concentration of reducing sugars in barley grain, but not alfalfa hay. Ruminal cellulolytic bacteria were more numerous with EF than with C. Disappearance of DM from barley grain was higher with EF than with C, but alfalfa was unaffected by EF. Treatment EF increased incorporation of 15N into FPA and FPB fractions at 24 and 48 h. In contrast, AEF reduced the 24 h values, relative to C; AEF and C were similar at 48 h. Infused enzyme (EI) did not affect 15N incorporation. Xylanase activity in effluent was increased by EF and EI, compared to C, but not by AEF. Xylanase activity in FPA was higher at 48 h than at 24 h with all treatments; it was higher with EF than C at 24 and 48 h, but was not altered by AEF or EI. Applying enzymes onto feeds before feeding was more effective than dosing directly into the artificial rumen for increasing ruminal fibrolytic activity.

Fermentation characteristics and ruminal ciliate protozoal populations in cattle fed medium- or high-concentrate barley-based diets.

Fermentation characteristics were measured and numbers and distribution by genera of ciliate protozoa were determined in ruminal fluid samples collected from 10 ruminally cannulated steers during the first 30 d of their being fed barley-based diets containing 62% (Medium Barley) or 95% (High Barley) barley grain (DM basis). Ruminal samples were collected at 5-d intervals over the 30-d periods beginning after adaptation (i.e., at the first full feeding of each diet). Ruminal pH and ammonia concentrations were lower (P < 0.001) with the High Barley than with the Medium Barley diet. Concentrations of total VFA and propionate and amylase activity of ruminal fluid were higher (P < 0.001) on High Barley than on Medium Barley. Total protozoal numbers in ruminal fluid were 42% lower (P < 0.05) on High Barley (470 x 10(3)/mL) than on Medium Barley (804 x 10(3)/mL). On Medium and High Barley diets, respectively, Entodinium spp. made up 89 and 91% of the ciliate protozoal populations. With the Medium Barley diet, relative proportions of Dasytricha, Ophryoscolex, Ostracodinium, Diplodinium, and Metadinium spp. in the total ciliate population were 4.5, 0.4, 0.5, 0.7, and 0.3%, respectively. When the High Barley diet was fed, these genera were not detected. In a subsequent survey, ruminal samples were collected from 200 finishing cattle at slaughter. Average protozoal population was 328 x 10(3)/mL, and Entodinium spp. constituted 97% of the total. These data demonstrate that a large population of Entodinium spp. can persist in the rumen of cattle fed high barley grain-based finishing diets.

Barley processing, forage:concentrate, and forage length effects on chewing and digesta passage in lactating cows.

Dietary factors that alter fermentability, NDF content, or particle size of the diet were evaluated for their effects on chewing behavior and distribution and passage of feed particles in the digestive tract of dairy cows. A double 4 x 4 quasi-Latin square design with a 2(3) factorial arrangement of treatments was used. The dietary factors were: extent of barley grain processing, coarse (1.60 mm) or flat (1.36 mm); forage-to-concentrate ratio (F:C), low (35:65) or high (55:45) (dry matter basis); and forage particle length, long (7.59 mm) or short (6.08 mm). Eight lactating cows with ruminal and duodenal cannulas were offered ad libitum access to total mixed diets. Chewing time, expressed as minutes per day or per kilogram of dry matter or neutral detergent fiber (NDF), was increased with high F:C diets due to increased eating and ruminating times but was decreased when expressed per kilogram of NDF intake from forage. The influence of forage particle length or grain processing on chewing activity was less pronounced than F:C ratio. Chewing activity was positively correlated to proportion of long forage particles in the diet but not to particle length of the diets. Influence of feed particle size on particle size distribution in different sites of the digestive tract was minimal. Particle size distributions of duodenal digesta and feces differed; the proportion of particles retained on the 3.35- or 1.18-mm screens was higher, but proportion of particles that passed through the 1.18-mm screen was lower in duodenal digesta than in feces. Relationships between chewing activities and ruminal pH or fractional passage rate of rumen contents were not significant. These results indicate that particle size of barley-based diets was not a reliable indicator of chewing activity. Forage particle size and NDF content of the diets were more reliable indicators of chewing activity than was the NDF content of forage. Fecal particle size was not an appropriate means of estimating the size of particles exiting the reticulorumen, at least for barley-based diets. Breakdown of coarse particles was necessary, but not a rate-limiting step for particles exiting the rumen. Passage rate of the rumen contents was affected by numerous factors including chewing activity.

Comparison of the ruminal metabolism of nitrogen from 15N-labeled alfalfa preserved as hay or as silage.

Alfalfa (Medicago sativa L. cv. AC Blue J) was labeled with 15N during growth in a greenhouse, harvested at early bloom, and preserved as silage (19% dry matter) or as sun-cured hay. The labeled silage and hay were given as single-pulse doses to two lactating Holstein cows fed diets comprising 30% concentrate and 70% alfalfa forage (preserved either as silage or as hay). Labeled forage and ruminal content samples collected for 72 h after dosing were partitioned into N fractions and analyzed for 15N-enrichment. Pool sizes of N compartments and kinetics in the rumen were derived by isotope dilution and by gravimetric measurements. The rate of outflow of total N, determined gravimetrically, was 21% higher with the silage diet than with the hay diet. On both diets, the largest individual flux was associated with the nonprotein, nonammonia, nonmicrobial nitrogen (NPAM-N) pool. As related to the flux of 15N through the acid detergent insoluble N pool, less tracer passed through the solid-phase nonfiber N and the soluble protein-N pools, and more passed through the NPAM-N pool, with silage than with hay. The solid-phase nonfiber N pool, which includes readily available feed N and adherent bacterial- and protozoal-N, constituted the largest N entity in the rumen, followed by the NPAM-N pool. When the forage component of the diet was alfalfa silage, N flux through the NPAM-N pool was remarkably high, and with both methods of preserving alfalfa forage, the exchange of tracer was most intensive through this pool.