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Influence of fibrolytic enzymes on the hydrolysis and fermentation of pure cellulose and xylan by mixed ruminal microorganisms in vitro.

A series of in vitro studies was conducted to determine the effects of adding a commercial enzyme product on the hydrolysis and fermentation of cellulose, xylan, and a mixture (1:1 wt/wt) of both. The enzyme product (Liquicell 2500, Specialty Enzymes and Biochemicals, Fresno, CA) was derived from Trichoderma reesei and contained mainly xylanase and cellulase activities. Addition of enzyme (0.5, 2.55 and 5.1 microL/g of DM) in the absence of ruminal fluid increased (P < 0.001) the release of reducing sugars from xylan and the mixture after 20 h of incubation at 20 degrees C. Incubations with ruminal fluid showed that enzyme (0.5 and 2.55 microL/g of DM) increased (P < 0.05) the initial (up to 6 h) xylanase, endoglucanase, and beta-D-glucosidase activities in the liquid fraction by an average of 85%. Xylanase and endoglucanase activities in the solid fraction also were increased (P < 0.05) by enzyme addition, indicating an increase in fibrolytic activity due to ruminal microbes. Gas production over 96 h of incubation was determined using a gas pressure measurement technique. Incremental levels of enzyme increased (P < 0.05) the rate of gas production of all substrates, suggesting that fermentation of cellulose and xylan was enzyme-limited. However, adding the enzyme at levels higher than 2.55 microL/g of DM failed to further increase the rate of gas production, indicating that the maximal level of stimulation was already achieved at lower enzyme concentrations. It was concluded that enzymes enhanced the fermentation of cellulose and xylan by a combination of pre- and postincubation effects (i.e., an increase in the release of reducing sugars during the pretreatment phase and an increase in the hydrolytic activity of the liquid and solid fractions of the ruminal fluid), which was reflected in a higher rate of fermentation.

Animal Feed↗

Soybean oil supplementation of a high-concentrate diet does not affect site and extent of organic matter, starch, neutral detergent fiber, or nitrogen digestion, but influences both ruminal metabolism and intestinal flow of fatty acids in limit-fed lambs.

Our objective was to measure ruminal fermentation characteristics and site and extent of nutrient digestion in sheep limit-fed an 81.6% (DM basis) concentrate diet supplemented with increasing levels of soybean oil. Eight white-faced wether lambs (39.9+/-3.0 kg BW) fitted with ruminal, duodenal, and ileal cannulas were used in a replicated 4 x 4 Latin square experiment. Diets were formulated to contain 15.0% CP (DM basis) and included bromegrass hay (18.4%), cracked corn, soybean oil, corn gluten meal, urea, and limestone. Soybean oil was added to diets at 0, 3.2, 6.3, and 9.4% of dietary DM. The diet was limit-fed at 1.4% of BW. After 14 d of dietary adaptation, Cr2O3 (2.5 g) was dosed at each feeding for 7 d followed by ruminal, duodenal, ileal, and fecal sample collections for 3 d. Digestibilities of OM, starch, NDF, and N were not affected (P = 0.13 to 0.95) by increasing dietary soybean oil level. Means for true ruminal (percentage of intake), lower-tract (percentage entering the duodenum), and total-tract (percentage of intake) digestibility for each nutrient were (mean+/-SEM): OM = 50.7+/-4.66%, 71.6+/-2.58%, and 82.7+/-0.93%; starch = 92.0+/-1.94%, 96.1+/-0.70%, and 99.8+/-0.05%; NDF = 36.7+/-6.75%, 50.9+/-7.58%, and 71.7+/-1.93%; and N = 31.6+/-9.93%, 84.1+/-1.50%, and 81.0+/-1.10%, respectively. Total VFA concentration was greatest in sheep fed 6.3% soybean oil and least in sheep fed 9.4% soybean oil (cubic, P = 0.01). Duodenal flow of fatty acids from the diet and those metabolized within the rumen increased (linear, P < 0.001) with increasing dietary soybean oil level. Ileal flow of 16:0, 17:0, 18:0, 18:1trans, and 18:1cis-9 fatty acids increased (P < or = 0.04) with increasing dietary soybean oil level. Apparent small intestinal disappearance of 18:0 decreased (linear, P = 0.004) as dietary soybean oil increased, and with 9.4% dietary soybean oil, nearly half the duodenal 18:0 was observed at the ileum; thus, the true energy value of the soybean oil decreased with increasing oil supplementation. We conclude that supplementation of a high-concentrate diet with increasing amounts of soybean oil in limit-fed sheep resulted in a trade off between loss of potential dietary energy from the fat and gain of important PUFA and biohydrogenation intermediates, but without a marked influence on digestibility of other important macronutrients.

Animal Feed↗

Ruminal fermentation and degradation patterns, protozoa population, and urinary purine derivatives excretion in goats and wethers fed diets based on two-stage olive cake: effect of PEG supply.

Three experiments were conducted in Granadina goats and Segureña wethers fed at maintenance level to evaluate the effect of including a mixture of barley and a new by-product derived from olive oil extraction (two-stage dried olive cake) on ruminal degradation and passage kinetics (Exp. 1), fermentation pattern and protozoa population (Exp. 2), and urinary purine derivatives excretion (Exp. 3). Polyethylene glycol was supplied to the animals to evaluate the effects of tannins contained in the by-product. The experimental diets were as follows: alfalfa hay and alfalfa hay plus a concentrate, formulated with two-stage dried olive cake, barley, and a mineral-vitamin mixture either with or without the addition of polyethylene glycol to the drinking water. The inclusion of two-stage dried olive cake in the diet resulted in an increase of condensed tannins. Ruminal VFA concentration in goats and wethers increased (P < 0.05) and ammonia N (NH3-N) concentration decreased (P < 0.05). The inclusion of two-stage dried olive cake decreased (P < 0.001) urinary allantoin excretion only in wethers. Ruminal degradation profiles and fractional passage rates were similar in goats and wethers. The polyethylene glycol supply increased (P < 0.001) DM and N degradation rates in both animal species but did not modify the fractional passage rate. Ruminal fermentation patterns were also similar in goats and wethers and were affected by polyethylene glycol supply. In general, Entodiniomorphida and Holotricha protozoa counts were higher (P < 0.05) in the rumen of goats than of wethers. Protozoa count in wethers responded more to polyethylene glycol supply than in goats. The present work presents the first data obtained from a comparative study with sheep and goats concerning urinary excretion of purine derivatives. The excretion was similar in both animal species when fed alfalfa hay; however, polyethylene glycol affected only urinary allantoin excretion in wethers. Results suggest a greater sensitivity of wethers than of goats to two-stage olive cake condensed tannins.

Ammonia↗

Evaluation of perennial ryegrass straw as a forage source for ruminants.

Two experiments were conducted to evaluate perennial ryegrass straw as a forage source for ruminants. Experiment 1 evaluated digestion and physiological variables in steers offered perennial ryegrass straw containing increasing levels of ergot alkaloid, lolitrem B. Sixteen ruminally cannulated Angus x Hereford steers (231+/-2 kg BW) were blocked by weight and assigned randomly to one of four treatments. Steers were provided perennial ryegrass straw at 120% of the previous 5-d average intake. Before straw feeding, soybean meal was provided (0.1% BW; CP basis) to meet the estimated requirement for degradable intake protein. Low (L) and high (H) lolitrem B straws (<100 and 1,550 ppb, respectively; DM basis) were used to formulate treatment diets: 100% L; 67% L:33% H; 33% L:67% H; 100% H (DM basis). Intake and digestibility of DM and OM, and ruminal pH, total VFA, and NH3-N were not affected by increasing lolitrem B concentration. Ruminal indigestible ADF (IADF) fill increased linearly (P = 0.01) and IADF passage rate decreased linearly (P = 0.04) as lolitrem B increased. Experiment 2 evaluated performance and production by 72 Angus x Hereford cows (539+/-5 kg BW) consuming perennial ryegrass straw containing increasing lolitrem B during the last third of gestation. Cows were blocked by body condition score and randomly assigned to one of three treatments. Cows were provided perennial ryegrass straw ad libitum and supplemented with soybean meal (0.1% BW; CP basis) to meet the estimated requirement for degradable intake protein. Mixtures of a L and H lolitrem B straw (467 and 2,017 ppb, respectively) were used to formulate treatment diets: 100% L, 50% L:50% H, 100% H (DM basis). Thirteen of 24 cows on the 100% H treatment exhibited signs of ryegrass staggers and were removed from the study; nevertheless, lolitrem B concentration did not influence pre- or postcalving weight or body condition score change. These data suggest that feeding perennial ryegrass straw containing up to 1,550 ppb lolitrem B (DM basis) did not adversely affect nutrient digestion or physiological response variables in steers. However, providing straw with a lolitrem B concentration of approximately 2,000 ppb (DM basis) resulted in 54% of cows exhibiting signs of ryegrass staggers. These data suggest that blending straws with a high (>2,000 ppb) and low (<500 ppb) concentration of lolitrem B can be a successful management practice.

Animal Feed↗

Effect of energy source and ruminally degradable protein addition on performance of lactating beef cows and digestion characteristics of steers.

Two trials were conducted to determine the effect of energy source (ENG) and ruminally degradable protein (RDP) on lactating cow performance and intake and digestion in beef steers. In Trial 1, 78 cow-calf pairs were used in a 2 x 2 factorial design to determine the effect of ENG (corn or soyhulls; SH) and RDP (with our without sunflower meal) to a forage diet for lactating beef cows. The basal diet consisted of 75% grass hay (11.5% CP) and 25% wheat straw (7.4% CP). Supplement treatments and predicted RDP balances were corn (-415 g of RDP/d); SH (-260 g of RDP/d); corn plus RDP (0 g of RDP/d); or SH plus RDP (0 g of RDP/d). Data were analyzed as a split-plot in time, with pen as the experimental unit (two pens per treatment). No interaction between ENG and RDP was present (P > 0.08) for any response variable. No differences (P > 0.39) due to ENG or RDP were noted for BW, BCS, or milk yield; however, final calf weight tended to increase with ENG (P = 0.06). In Trial 2, a 5 x 5 Latin square was used to determine effects of ENG and RDP on intake and digestion in steers (686 +/- 51 kg BW). Treatments were arranged as a 2 x 2 plus one factorial and comprised a control (CON; grass hay, 7% CP), grass hay plus 0.4% BW SH, grass hay plus 0.4% BW SH and 0.15% BW sunflower meal, grass hay plus 0.4% BW corn, and grass hay plus 0.4% BW corn and 0.2% BW sunflower meal. Preplanned contrasts included main effects of ENG and RDP, ENG x RDP interaction, and CON vs. supplemented (SUP) treatments. Supplementation increased total DMI compared with CON (P = 0.001), but forage DMI was greater (P = 0.001) for CON than for SUP. An ENG x RDP interaction occurred for forage DMI (P = 0.02); addition of RDP to corn decreased forage intake, whereas addition of RDP to SH had no effect. There was an ENG x RDP interaction (P = 0.001) for ruminal pH; pH tended to increase with RDP addition to SH (P = 0.07), but decreased with RDP addition to corn (P = 0.001). Supplementation increased ruminal ammonia compared with CON (P = 0.001). Likewise, RDP increased ruminal ammonia (P = 0.001). An interaction occurred for OM disappearance (OMD; P = 0.01). The RDP addition to SH numerically decreased OMD (P = 0.23), whereas RDP addition to corn numerically increased OMD (P = 0.14). Intake and digestion seem to respond differently to RDP addition depending on supplemental energy source. Both corn or SH seem to be suitable supplements for the quality of forage used in this trial. Addition of supplemental protein did not improve cow or calf performance.

Animal Feed↗

Ruminal ammonia load affects leucine utilization by growing steers.

Six ruminally cannulated Holstein steers (initial BW = 189 +/- 11 kg) housed in metabolism crates were used in a 6 x 6 Latin square to study effects of ruminal ammonia load on Leu utilization. All steers received a diet based on soybean hulls (2.7 kg of DM/d), ruminal infusions of 200 g of acetate/d, 200 g of propionate/d, and 50 g of butyrate/d, as well as an abomasal infusion of 300 g of glucose/d to provide energy without increasing microbial protein supply and an abomasal infusion of a mixture (238 g/d) of all essential AA except Leu. Treatments were arranged as a 3 x 2 factorial and included Leu (0, 4, or 8 g/d) infused abomasally and urea (0 or 80 g/d) infused ruminally. Abomasal Leu infusion linearly decreased (P < 0.05) both urinary and fecal N excretions and linearly increased (P < 0.05) retained N, but the decreases in urinary N excretion in response to Leu tended (P = 0.07) to be greater, and the increases in retained N in response to Leu were numerically greater in the presence of the urea infusion. Although urea infusions increased (P < 0.05) plasma urea concentrations, urinary N excretions, and urinary urea excretions, retained N also was increased (P < 0.05). The efficiency of deposition of supplemental Leu ranged from 24 to 43% when steers received 0 or 80 g of urea/d, respectively. Under our experimental conditions, increasing ammonia load improved whole-body protein deposition in growing steers when Leu supply was limiting.

Abomasum↗

Effect of diets containing linoleic acid- or oleic acid-rich oils on ruminal fermentation and nutrient digestibility, and performance and fatty acid composition of adipose and muscle tissues of finishing cattle.

Two trials were conducted to determine the effect of linoleic acid- or oleic acid-rich safflower oil on ruminal fermentation, nutrient digestion, feedlot performance, carcass characteristics, and fatty acid composition of adipose and muscle tissues of beef cattle. In both trials, cattle were fed a finishing diet based on barley grain, wheat silage, and alfalfa hay. Oils were fed at 5% of dietary DM. In a metabolism trial, four ruminally and duodenally cannulated Angus crossbred steers were subjected to linoleic acid-rich oil or oleic acid-rich oil in a crossover design with covariate periods (no oil supplementation). In a finishing trial, 16 individually fed Angus crossbred steers and heifers (eight per diet) received linoleic acid- or oleic acid-rich oils during the last 86 d of a 116-d feeding period. Ruminal pH, ammonia concentration, protozoal counts, major VFA concentrations, acetate-to-propionate ratio, polysaccharide-degrading activities, microbial N flow to the duodenum, and the efficiency of microbial N synthesis in the rumen were not affected (P = 0.18 to 0.96) by type of oil. Type of oil had no effect on total-tract apparent digestion of nutrients (P = 0.46 to 0.98). Ruminal true nutrient digestibilities did not differ between oils (P = 0.15 to 0.99), except that the linoleic acid-rich oil decreased (P = 0.05) NDF digestibility. Dry matter intake, ADG, G:F, and carcass characteristics did not differ (P = 0.11 to 0.84) between the two oils. Overall, the difference in dietary fatty acids provided to the cattle produced few changes in tissue fatty acids. Weight percentages of c9t11 CLA were unaltered by the addition of linoleic acid to the diet compared with oleic acid, probably as a result of low vaccenic acid production in the rumen, as the pathway of biohydrogenation was apparently primarily through the t10 pathway.

Adipose Tissue↗

Changes in ruminal fermentation and protein degradation in growing Holstein heifers from 80 to 250 kg fed high-concentrate diets with different forage-to-concentrate ratios.

Six Holstein heifers (initial BW = 65.2 +/- 1.8 kg) fitted with ruminal cannulas were used in a repeated measures trial to assess the effect of age and forage-to-concentrate ratio on ruminal fermentation end products and in situ degradation kinetics of four plant protein supplements (soybean meal, sunflower meal, peas, and lupin seeds). Alfalfa hay also was incubated in situ to estimate NDF degradation. Three experimental periods were conducted at 13, 27, and 41 wk of age. Heifers were fed one of two diets, 12:88 vs. 30:70 forage-to-concentrate ratio (DM basis), offered as total mixed ration on an ad libitum basis. Intakes of DM, OM, CP, NDF, and ADG were not affected (P > or = 0.105) by diet. The 30:70 diet resulted in faster (P = 0.045) fluid passage rate and decreased (P = 0.015) ammonia N concentration compared with the 12:88 diet, but no differences (P > or = 0.244) were detected in ruminal pH and total VFA concentration between diets. The rate of degradation and the effective degradability of N in protein supplements was greater with the 30:70 diet for peas (P < or = 0.008) and lupin seeds (P < or = 0.02), and in the 12:88 diet for sunflower meal (P < or = 0.06). Degradation of NDF of alfalfa hay was low with both diets (18.5 and 23.7 % for 12:88 and 30:70, respectively); however, the rate and extent of DM and NDF degradation were greater (P < or = 0.016) with the 30:70 diet, suggesting a higher cellulolytic activity. Total VFA concentration and the proportion of propionate increased (P < or = 0.035), and the acetate proportion decreased (P = 0.021) with age. Average pH, ammonia N concentration, and passage rates were not affected (P > or = 0.168) by age. Degradation rate and effective degradability of N of sunflower meal, peas, lupin seeds, and of DM of alfalfa hay increased (P < or = 0.08) with age, but degradation kinetics of NDF of alfalfa hay was not affected (P > or = 0.249). The increase in the rate and extent of N degradation with age would suggest an increase in proteolytic activity, and the changes in the fermentation pattern may reflect an increase in amylolytic activity caused mainly by an increase in the gross intake of nonstructural carbohydrates and by adaptation of ruminal microflora after long exposure to these nutrients.

Acetates↗

Supplementing a ruminally undegradable protein supplement to maintain essential amino acid supply to the small intestine when forage intake is restricted in beef cattle.

Twelve Angus crossbred cattle (eight heifers and four steers; average initial BW = 594 +/- 44.4 kg) fitted with ruminal and duodenal cannulas and fed restricted amounts of forage plus a ruminally undegradable protein (RUP) supplement were used in a triplicated 4 x 4 Latin square design experiment to determine intestinal supply of essential AA. Cattle were fed four different levels of chopped (2.54 cm) bromegrass hay (11.4% CP, 57% NDF; OM basis): 30, 55, 80, or 105% of the forage intake required for maintenance. Cattle fed below maintenance were given specified quantities of a RUP supplement (6.8% porcine blood meal, 24.5% hydrolyzed feather meal, and 68.7% menhaden fish meal; DM basis) designed to provide duodenal essential AA flow equal to that of cattle fed forage at 105% of maintenance. Experimental periods lasted 21 d (17 d of adaptation and 4 d of sampling). Total OM intake and duodenal OM flow increased linearly (P < 0.001) as cattle consumed more forage; however, OM truly digested in the rumen (% of intake) did not change (P = 0.43) as intake increased. True ruminal N degradation (% of intake) tended (P = 0.07) to increase linearly, and true ruminal N degradation (g/d) decreased quadratically (P = 0.02) as intake increased from 30 to 105%. Duodenal N flow was equal (P = 0.33) across intake levels, even though microbial N flow increased linearly (P < 0.001) as forage OM intake increased. Total and individual essential AA intake decreased (cubic; P < 0.001) as forage intake increased because the supply of nonammonia, nonmicrobial N flow from RUP was decreased (linear; P < 0.001) by design. Total duodenal flow of essential AA did not differ (P = 0.39) across these levels of forage intake. Although the profile of essential AA reaching the duodenum differed (P < or = 0.02) for all 10 essential AA, the range of each essential AA as a proportion of total essential AA was low (11.1 to 11.2% of total essential AA for phenylalanine to 12.3 to 14.3% of total essential AA for lysine). Duodenal essential AA flow did not differ (P = 0.10 to 0.65) with forage intake level for eight of the 10 essential AA. Duodenal flow of arginine decreased linearly (P = 0.01), whereas duodenal flow of tryptophan increased linearly (P = 0.002) as forage intake increased from 30 to 105% of maintenance. Balancing intestinal essential AA supply in beef cattle can be accomplished by varying intake of a RUP supplement.

Amino Acids↗

Effects of nonstructural carbohydrates and protein sources on intake, apparent total tract digestibility, and ruminal metabolism in vivo and in vitro with high-concentrate beef cattle diets.

To investigate the effects of synchronizing nonstructural carbohydrate (NSC) and protein degradation on intake and rumen microbial fermentation, four ruminally fistulated Holstein heifers (BW = 132.3 +/- 1.61 kg) fed high-concentrate diets were assigned to a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments studied in vivo and in vitro with a dual-flow continuous culture system. Two NSC sources (barley and corn) and 2 protein sources [soybean meal (SBM) and sunflower meal (SFM)] differing in their rate and extent of ruminal degradation were combined resulting in a synchronized rapid fermentation diet (barley-SFM), a synchronized slow fermentation diet (corn-SBM), and 2 unsynchronized diets with a rapidly and a slowly fermenting component (barley-SBM, and corn-SFM). In vitro, the fermentation profile was studied at a constant pH of 6.2, and at a variable pH with 12 h at pH 6.4 and 12 h at pH 5.8. Synchronization tended to result in greater true OM digestion (P = 0.072), VFA concentration (P = 0.067), and microbial N flow (P = 0.092) in vitro, but had no effects on in vivo fermentation pattern or on apparent total tract digestibility. The NSC source affected the efficiency of microbial protein synthesis in vitro, tending to be greater (P = 0.07) for barley-based diets, and in vivo, the NSC source tended to affect intake. Dry matter and OM intake tended to be greater (P > or = 0.06) for corn- than barley-based diets. Ammonia N concentration was lower in vitro (P = 0.006) and tended to be lower in vivo (P = 0.07) for corn- than barley-based diets. In vitro, pH could be reduced from 6.4 to 5.8 for 12 h/d without any effect on ruminal fermentation or microbial protein synthesis. In summary, ruminal synchronization seemed to have positive effects on in vitro fermentation, but in vivo recycling of endogenous N or intake differences could compensate for these effects.

Animal Feed↗

Inhibition of ruminal degradation of L-tryptophan to 3-methylindole, in vitro.

Available evidence supports the view that acute bovine pulmonary edema and emphysema (ABPE) is related to ruminal production of 3-methylindole (3MI) from L-tryptophan (TRP). Ruminal production of 3MI is a two-step process involving the conversion of TRP to indoleacetic acid (IAA) followed by decarboxylation of IAA and 3MI. Reduction in ruminal 2MI production by the inhibition of either of these processes may prevent the onset of ABPE. A closed-system, in vitro ruminal fermentation technique was used to screen 27 compounds for their ability to reduce the conversion of TRP to 3MI. Several compounds tended to reduce 3MI production at both 25 and 5 micrograms/ml. Desoxysalinomycin, X-206, chloral hydrate, nigericin, lasalocid, monensin, narasin and salinomycin all reduced 3MI production by more than 80% at 5 micrograms/ml without reducing total VFA production. All of these compounds, except chloral hydrate, are polyether antibiotics. At least part of the inhibition due to monensin and narasin occurs at the level of TRP conversion to IAA.

Animals↗

Nitrogen utilization and ruminal fermentation in steers fed soybean meal treated with formaldehyde.

Four rumen-fistulated steers averaging 400 kg in body weight were used in a 4 X 4 Latin square arrangement with 18-d periods to investigate the effect of treating soybean meal (SBM) with formaldehyde on nitrogen (N) utilization and ruminal fermentation. Experimental diets, on a dry matter basis, consisted of 42% corn silage, 48.5% cracked corn-mineral mixture and 9.5% SBM treated with 0, .3, .6 or .9% formaldehyde by weight. Dry matter and organic matter digestibilities were not affected by treatment. Formaldehyde treatment of SBM resulted in a linear decrease in N digestibility (P less than .005) and urinary N excretion (P less than .01) and a quadratic increase (P less than .05) in N retention. The depression in apparent N digestibility was small when SBM was treated with .3% formaldehyde. This level of formaldehyde treatment also had little effect on in vitro enzymatic hydrolysis of SBM. Ruminal ammonia-N concentrations were lower (P less than .05) in steers fed formaldehyde-treated SBM. Ruminal pH was lower (P less than .05) at 6 and 8 h postfeeding while volatile fatty acid concentrations were higher (P less than .05) at 8 and 12 h postfeeding for steers fed untreated SBM. Propionic acid (mol/100 mol) decreased linearly (P less than .05) with increasing level of formaldehyde treatment. Urea-N concentrations in plasma were decreased (P less than .001) and plasma-free essential amino acid concentrations were increased (P less than .10) by formaldehyde treatment. Ruminal disappearance of N from polyester bags containing the SBM supplements was greatly reduced (P less than .005) by formaldehyde treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

N tau-methylhistidine--feed and duodenal content and ruminal degradation.

N tau-methylhistidine (NMH) concentrations were determined in acid hydrolysates of 12 common feedstuffs for ruminants, and duodenal digesta from steers fed alfalfa haylage and high-moisture corn. For six species of grass hays, NMH concentrations ranged from nondetectable (ND; less than .1 nmol/g dry matter) to .6 nmol/g dry matter. Two legume hays had NMH concentrations of .9 and 2.0 nmol/g dry matter, respectively. For corn silage, high-moisture corn, beet pulp and soybean meal, NMH concentrations were 1.3, 1.1, 1.5 and 2.2 nmol/g dry matter, respectively. Duodenal digesta NMH concentrations ranged from ND to 1.2 nmol/g dry matter. When pure NMH (1 mM) was incubated with rumen fluid in vitro for 8 h, only 14% was degraded while leucine (1 mM) was totally degraded. From the feedstuffs used in this study, results show that exogenous (dietary) NMH is not a concern in skeletal muscle protein turnover studies in cattle using urinary NMH excretion because it can be calculated that potential NMH intake from the preceding feedstuffs would be less than 1% of normal urinary excretion and that ruminal microbes are not a potential source of NMH. NMH was resistant to in vitro ruminal degradation. The in vitro ruminal NMH degradation results suggest that when feedstuffs containing at least 15 nmol/g dry matter are fed (far greater than values observed in this study) a significant proportion of urinary NMH would arise directly from exogenous sources and would invalidate the use of this technique in assessing protein turnover in cattle.

Animal Feed↗

Ruminal changes in monensin- and lasalocid-fed cattle grazing bloat-provocative alfalfa pasture.

Microbial and fermentation changes in the rumen in monensin- and lasalocid-fed cattle grazing bloat-provocative alfalfa pasture were studied using genetically bloat-susceptible, ruminally-cannulated adult cattle. Monensin at .66 and .99 mg/kg body weight daily reduced the severity of legume bloat by 41 and 73%, respectively. The same doses of lasalocid reduced bloat by 25 and 12%. Comparison of ruminal contents from animals before treatment with ruminal contents from antibiotic-treated animals showed no differences in pH, ammonia, soluble N, soluble carbohydrate, ethanol-precipitable slime and anaerobic bacterial counts. Monensin treatment decreased protozoal numbers and microbial activity, as evidenced by lower gas production from in vitro fermentation of ground alfalfa hay when compared to pretreatment. Lasalocid had no effect on protozoal counts and in vitro gas production. Addition of monensin or lasalocid (12 micrograms/ml) to in vitro fermentation of chopped, fresh alfalfa reduced microbial activity as evidenced by higher soluble N, lower ammonia concentration and decreased gas production. Monensin reduced the amount of ethanol-precipitable slime and protozoal numbers. Reduction in the severity of bloat when monensin was fed appears to be due to decreased protozoal numbers, which resulted in decreased gas production. Lasalocid did not reduce legume bloat because of its minimal effect on the ruminal protozoa.

Animal Feed↗

Changes in ruminal concentrations of microbial ammonia and amino acids due to monensin and time.

The effect of monensin (33 mg/kg) and no monensin on distribution of ammonia and free amino acids in fluid and particulate phases of ruminal contents was studied. Two Jersey cows and two Angus-Hereford steers, weighing approximately 350 kg and each containing a ruminal cannula were meal fed a 65% concentrate diet for two periods. Feed was available for 30 min; any feed not eaten was removed after this time. Whole ruminal contents were sampled before feeding, immediately after feed removal and 1, 2, 3, 4, 6, 8, 10 and 12 h after feed removal. A crossover design with split-plot in time was used with four observations per time point per treatment. Extracellular (E) ammonia and amino acids were determined on high speed supernatants of strained ruminal fluid. Total (T) ammonia and amino acids were obtained by lysing microbial cells with 1% cetyltrimethylammonium bromide (CTAB) and one freeze treatment before centrifugation. Intracellular (I) amino acids and ammonia were determined as T minus E. Squeezed particles were suspended in .1 N HCl containing 1% CTAB. Total and E ammonia were higher throughout the study for no monensin vs monensin (P less than .05). Intracellular ammonia was low (.03 to 1 mM) and did not change (P greater than .10) across time for either dietary treatment. Intracellular amino acids ranged from 1.2 to 4.5 mM and were higher (P less than .05) than E amino acids (.9 to 2.2 mM) for both diets immediately after feeding. Monensin did not significantly lower (P greater than .10) T amino acids compared with no monensin throughout sampling (2.6 and 3.1 mM, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

Site and extent of neutral detergent fiber digestion, efficiency of ruminal digesta flux and fecal output as related to variations in voluntary intake and chewing behavior in heifers.

Six heifers (1/4 Brahman x 1/4 Jersey x 1/2 Angus, 290 kg avg wt) with ruminal, duodenal and ileal cannulas were given ad libitum access to a corn silage-based diet with or without 100 mg monensin.hd-1.d-1 in a two-period crossover design. There was no significant effect of monensin on voluntary intake (VI), flow of NDF to the abomasum, fecal output or the efficiency of transfer of NDF to the duodenum (unitary fluxes of NDF through the reticulo-omasal orifice, number.g NDF.(kg BW.75)-1). Monensin increased extent of NDF digestion (NDFD) in the dorsal (P = .07) and ventral (P = .09) ruminal sac. As a fraction of the total tract NDFD, NDFD increased progressively in dorsal rumen (.214), ventral rumen (.548) and duodenal (.818) sites. The VI among individual heifers was negatively related to unitary fluxes of NDF through the reticulo-omasal orifice (P = .001) and to the extent of digestion of potentially digestible NDF (PDNDFD) in the ventral rumen (P = .006) and duodenum (P = .032), but not in the dorsal rumen (P = .720) or in the total tract (P = .785). Fecal output of NDF was negatively related to unitary times of eating (P = .027), ruminating (P = .001), masticating (P = .001) and unitary number of rumination boli (P = .001). These relationships all were influenced (P = .001) by individual heifer.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of supplemental potassium and sodium chloride salts on ruminal turnover rates, acid-base and mineral status of lactating dairy cows during heat stress.

Effects of added dietary sodium and potassium chloride salts on ruminal turnover rates, acid-base balance and mineral status of lactating dairy cows experiencing a nycterohemeral cycle of heat stress were examined. Black globe-humidity index in the chambers averaged 94 during the daytime and 68 during the nighttime. Four ruminally cannulated multiparous Holstein cows in mid-lactation were confined in climatic chambers for a single-reversal experiment consisting of two 17-d periods. To the basal diet (50% corn silage: 50% concentrate, which contained .97% potassium, .19% sodium and .20% chloride), 1.25% sodium chloride plus 1.85% potassium chloride were added, making the high mineral treatment (1.93% potassium, 68% sodium and 1.85% chloride). Liquid dilution rates from the rumen were measured by chromium-ethylenediaminetetraacetate disappearance. Turnover rates of solids were determined by appearance of ytterbium in feces. Ruminal contents, arterial blood and urine were collected hourly for 26 h. Grab samples of feces were sampled over 6 d. Dry matter intakes and milk yields were not affected by the diets (averaging 17.8 and 21.1 kg/d, respectively). Cows fed the high mineral diet drank 17% more water (P less than .01). Tests for homogeneity of regression were utilized to compare chromium disappearance and ytterbium appearance data, which were best described by second-order polynomial functions. Increased ruminal chromium disappearance (P less than .01) and decreased total volatile fatty acid concentrations (P less than .01) suggested faster liquid dilution rates with high mineral diet, but turnover rates of solids were not affected. Urinary potassium secretion compensated for the high potassium content of the high mineral diet without an alkalogenic effect on acid-base status. Lower urine pH and higher urine ammonium concentrations during cool hours suggested that the high chloride content of the high mineral diet had an acidogenic effect. The results are interpreted to indicate that high level inclusion of sodium and potassium chloride salts altered digestive, acid-base and mineral status of heat-stressed lactating cows.

Acid-Base Equilibrium↗

Feed intake and digestion by beef cows fed prairie hay with different levels of soybean meal and receiving post-ruminal administration of antibiotics.

Six cannulated beef cows (one Angus, two Hereford and three Angus x Hereford; 405 kg) were used in a 6 x 6 latin square experiment with a 2 x 3 factorial arrangement of treatments. Prairie hay (.77% N, 73% neutral detergent fiber [NDF] and 7% acid detergent lignin) was fed ad libitum from d 1 through 14 and at 90% of ad libitum intake from d 15 through 21 during digesta collection. Periods lasted 21 d. Soybean meal (SBM) was offered at 0 (control, C), .12 (low, L) or .24% of body weight (high, H; dry matter basis). Cows received daily doses of an antibiotic mixture (1 g neomycin and .125 g bacitracin) or saline in the duodenum. Prairie hay dry matter (DM) intake increased (P less than .05) linearly with SBM supplementation, being 25 and 40% greater for L and H than for C, respectively. Ruminal fluid concentrations of NH3-N and total volatile fatty acids increased (P less than .05) linearly as SBM was added to the diet. Ruminal fluid dilution rate increased linearly and particulate passage rate increased (P less than .05) quadratically with increasing SBM. True ruminal digestibilities of organic matter, NDF and N increased (P less than .10) quadratically with increasing SBM (organic matter; 50.3, 57.9 and 58.3%; NDF: 54.7, 60.4 and 59.8%; N: 17.5, 45.1 and 51.4% for C, L and H, respectively). Main effects of antibiotic administration were not significant. Increases in DM intake when SBM was given were large compared with the small elevations in ruminal digestion, implying that metabolic regulation was modifying intake of low-quality forage.

Animal Feed↗