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The effect of monensin supplementation on ruminal ammonia accumulation in vivo and the numbers of amino acid-fermenting bacteria.

When nonlactating Holstein cows (685 +/- 59 kg) were fed chopped timothy hay (9% CP, 7.0 kg/d) 12 times daily, the steady-state ruminal ammonia concentration was 2.6 mM, and the specific activity of ammonia production by mixed ruminal bacteria was 27.4 nmol/mg of protein-1.min-1. The addition of soybean meal (53% CP, 1 or 2 kg/d) to the basal diet caused a linear increase in ruminal ammonia (7.0 and 12.4 mM, respectively; P < .001), but there was only a small increase in the specific activity of ammonia production (30.7 and 33.8 nmol/mg of protein-1.min-1, respectively; P < .05). The addition of monensin (350 mg/d) to the diets caused more than a 30% decrease (P < .01) in ruminal ammonia at all levels of soybean supplementation, and there was a similar decrease (P < .001) in the specific activity of ammonia production. Before monensin addition, the most probable number of bacteria that could utilize peptides and amino acids, but not carbohydrates, as an energy source for growth was 5.8 to 7.0 x 10(6)/mL. When monensin was added to the diets, these bacteria decreased (P < .001) nearly 10-fold. Based on these results, it seemed that monensin inhibited highly active amino acid-fermenting ruminal bacteria, and this inhibition, in turn, decreased ruminal amino acid deamination and ammonia production. Because monensin did not increase soluble protein, peptides, or amino acids in ruminal fluid, it did not seem that the decrease in ammonia increased flow of dietary amino N to the lower gut.(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonia↗

Characteristics of ruminal and total tract digestion of canola meal and soybean meal in a high-energy diet for feedlot cattle.

Four Holstein steers (233 kg) with ruminal cannulas and "T" cannulas in the proximal duodenum were used in a 4 x 4 Latin square experiment to evaluate characteristics of ruminal and total tract digestion of a generic (CM-40, 40% CP) and a high-protein (CM-46, 46% CP) variety of canola meal (CM) compared with a conventional, solvent-extracted soybean meal (SBM). Treatments consisted of an 80% concentrate diet that contained 20% tapioca (TP) or 20% of the respective test protein sources. Supplemental proteins replaced TP in the basal diet. Intake of dietary treatments was restricted to 5.0 kg/d (DM basis, 2.2% BW). Ruminal digestion of OM was similar (P > .10) for the CM-supplemented diets. Ruminal digestion of OM was higher (5.8%, P < .01) for SBM than for CM diets. Ruminal digestibility of ADF was similar (P > .10) across treatments. Ruminal degradation of feed N was similar (P > .10) for CM diets but tended to be greater (6.7%, P < .10) for SBM. Ruminal degradation of N in CM-40, CM-46, and SBM averaged 71, 73, and 80, respectively. Passage of individual amino acids to the small intestine was similar for CM and SBM diets. Total tract DM digestibility (DMD) was similar (P > .10) for the CM diets. Total tract DMD was higher (2.9%, P < .01) for SBM than for CM diets. Total tract digestibility of ADF also tended to be higher (14; P < .10) for SBM than for CM diets. The DE (megacalories/kilogram) of the CM and SBM diets were greater (P < .01) than for the TP diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Supplemental methionine and time of supplementation effects on ruminal fermentation, digesta kinetics, and in situ dry matter and neutral detergent fiber disappearance in cattle.

Two experiments were conducted to study the effects of methionine supplementation on ruminal fermentation and digesta kinetics. In Exp. 1, nine ruminally cannulated beef heifers (average initial BW = 527 kg) in a crossover design were fed low-quality grass hay and cottonseed meal with or without 11.4 g of supplemental methionine (polysaccharide-coated). Particulate and fluid kinetics, rate of DM and NDF disappearance, ruminal VFA and NH3 N concentrations, and pH were not altered (P > .10) by supplemental methionine; however, ruminal purine concentration was greater (P < .05) in methionine-supplemented heifers than in unsupplemented heifers. In Exp. 2, 12 ruminally cannulated Holstein steers (average initial BW = 622 kg) grazing a fescue pasture were allotted to one of three groups: no supplemental methionine (CON) or 11.4 g of supplemental methionine fed at 0700 (AM) or at 1200 (PM). Forage intake, particulate kinetics, ruminal fluid kinetics, pH, VFA, and NH3 N concentrations were not altered (P > .10) by supplemental methionine or supplementation time. In situ rate of DM and NDF disappearance was greater (P < .05) in supplemented steers than in CON steers; AM steers exhibited faster (P < .05) rates than PM steers. Overall, methionine supplementation of low-quality forage increased ruminal purine concentration but did not alter in situ fermentation or digesta passage, whereas supplementation at 0700, but not at 1200, of steers grazing fescue forage increased rate of NDF fermentation.

Ammonia↗

Net flux of metabolites across the ruminal wall of sheep fed twice a day with orchardgrass hay.

Four Texel wethers (68 +/- 2.5 kg BW) fitted with catheters in the ruminal veins and a mesenteric artery, blood flow probes around ruminal arteries, and a ruminal cannula were used to determine meal-related variations and daily significance of net flux across the ruminal wall of urea and ammonia (NH3), VFA, D-beta-hydroxybutyrate (D beta HOB), lactate, and glucose. Sheep were fed every 12 h with orchardgrass hay (430 g of DM/meal; 611 g of digestible OM/kg of DM and 23.6 g of N/kg of DM). Apart from lactate and glucose, the fluxes of studied metabolites were significantly affected by time after morning feeding. Maximum absorption of VFA and NH3 were observed at the end of the meal; however, 5 h after the meal VFA absorption was still high, whereas NH3 absorption had decreased to the prefeeding level. Net release of D beta HOB was greater during the 2 h after the meal than during the rest of the time. Urea net transfer decreased during the meal, and thereafter it increased to the 5th h after feeding, at which time it was twofold higher than at prefeeding. The difference in net flux across the ruminal wall of urea and NH3 was linearly correlated with NH3 concentration in the ruminal fluid. Daily urea and NH3 net transfer were -2.10 and 3.76 g of N/d, respectively. The VFA net appearance in the ruminal veins was 1.167, .226, and .014 mol/d for acetate, propionate, and butyrate, respectively. Daily net release of D beta HOB, lactate, and glucose by the rumen wall was .153, .093, and -.012 mol/d, respectively.

Absorption↗

Efficacy of laidlomycin propionate to reduce ruminal acidosis in cattle.

Three trials were conducted to evaluate the efficacy of laidlomycin propionate (LP) to reduce the incidence and severity of ruminal acidosis in cattle fed high-grain finishing diets. In each trial, LP was fed at 0, 6, or 12 mg/kg of diet DM. In two acidosis-challenge trials, ruminally fistulated steers were fed (DM basis) a 50% concentrate diet and then fed a 95% concentrate diet at a specific intake (2.75% BW) or steers were dosed intraruminally with a 100% concentrate diet. Laidlomycin propionate did not alter ruminal pH or total acid concentrations, but in Trial 1 the 6 mg/kg level altered (P < .10) the molar proportions of the acids, increasing total ruminal VFA and decreasing ruminal lactate. In Trial 3, a finishing trial, LP reduced (P < .10) intake day-to-day variation of individually fed steers during a 13-d adaptation period from a 65 to a 100% concentrate diet, suggesting reduced incidence of subacute acidosis. Feed intake was lower (P < .05) during the first 13 d of the trial due to LP but was not affected over the entire trial. Laidlomycin propionate improved feed efficiency (gain/feed) when calculated on a live weight basis (linear, P = .05) or carcass weight basis (linear, P = .20). Laidlomycin propionate does not prevent ruminal acidosis, but it may reduce the severity of ruminal acidosis during adaptation to a 100% concentrate diet.

Acidosis↗

Insulin and glucagon regulation of gluconeogenesis in preruminating and ruminating bovine.

Hepatocyte monolayers were established from young preruminating (7 to 14 d of age) or older ruminating (11 to 12 wk of age) calves and used to evaluate the effects of insulin and glucagon on incorporation of carbon from 2.5 mM [2-14C]propionate and 2.0 mM [U-14C]lactate into glucose and glycogen. Developmental state (young preruminating vs older ruminating) of the donor calf did not affect the rate of gluconeogenesis from propionate in the absence of hormones. Insulin decreased (P < .05) gluconeogenesis and increased (P < .05) glycogenesis from propionate and lactate in hepatocytes from preruminating calves but had no effect on hepatocytes from ruminating calves. Lactate was poorly metabolized to glucose and was not responsive to glucagon in hepatocytes from ruminating calves compared with hepatocytes from preruminating calves. Hepatocytes responded to glucagon by increasing (P < .05) gluconeogenesis from propionate. Maximal responsiveness to glucagon did not differ between ruminating and preruminating calves, but hepatocytes from preruminating calves responded at lower glucagon concentrations (P < .05). These data demonstrate a similar capacity of hepatocytes from preruminating and ruminating calves to metabolize propionate to glucose, but there was a seven- to eightfold decrease in gluconeogenesis from lactate in ruminating calves that was accompanied by a decreased response to acute changes in insulin and glucagon.

Animals↗

Effect of malate on in vitro mixed ruminal microorganism fermentation.

The objective of this study was to evaluate the effects of different concentrations of DL-malate (disodium salt) on the in vitro mixed ruminal microorganism fermentation of soluble starch or cracked corn. Ruminal fluid was collected from a steer fed 6.8 kg of forage and 2.3 kg of concentrate supplement once daily, and mixed ruminal microorganisms were incubated in anaerobic media (40 mL) that contained 20% (vol/vol) ruminal fluid in batch culture for 24 h at 39 degrees C. Malate was added to the incubation bottles (n = 4) to achieve final concentrations of 0, 4, 8, and 12 mM. When mixed ruminal microorganisms were incubated with only DL-malate as the substrate, final pH numerically increased, propionate and total VFA concentrations increased (P < .05), and the acetate:propionate ratio decreased (P < .05) as the concentration of DL-malate increased from 0 to 12 mM. Fermentation of cracked corn in the presence of 8 or 12 mM DL-malate resulted in an increase (P < .05) in final pH and propionate concentration. Total VFA tended to increase (P < .21), whereas final concentrations of L-lactate numerically decreased. In the case of soluble starch, 8 and 12 mM DL-malate caused a decrease (P < .05) in methane concentration. When only ruminal fluid (no added anaerobic medium) was used as the inoculum rather than 20% ruminal fluid medium, similar results for final pH, propionate, L-lactate, and total VFA were observed for soluble starch and corn incubations treated with DL-malate.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Propionate modulation of ruminal ketogenesis.

The ruminal epithelium is the primary source, through the metabolism of butyrate, of circulating ketone bodies in fed ruminants. Volatile fatty acid metabolism was investigated in short-term (2-h) incubations of isolated sheep ruminal epithelial cells. Ruminal epithelial cells were isolated from ruminal papillae via serial tryptic digestion. Cells were incubated in the presence of various combinations of butyrate (0, .5, 1, 5, 10, or 25 mM), propionate (0, .1, .5, 1, 5, 10, 25, 50, or 100 mM), acetate (0, .5, 1, 5, 25, 50, or 100 mM), and succinate (0, 5, 15, 50 mM) to evaluate the effects of these substrates on butyrate metabolism. Variables measured included beta-hydroxybutyrate (beta-HBA) and acetoacetate (AcAc) production from butyrate, [14C]butyrate oxidation to 14CO2, and lactate and pyruvate formation from propionate and succinate. Butyrate oxidation to CO2, beta-HBA and total ketones produced (nanomoles of butyrate metabolized per 10(6) cells per minute) were linear over the 2-h incubation period (y = .805X + 2.24, r2 = .78; y = 1.02X - 9.1, r2 = .72; and y = 2.238X - 11.76, r2 = .65, respectively). Acetate inhibited beta-HBA formation from butyrate (P < .05) when present at concentrations greater than 5 mM, although a dose-dependence was not consistently exhibited. However, butyrate oxidation to beta-HBA was stimulated (P < .05) by propionate while acetoacetate production declined (P < .05), resulting in no net change in ketone body production (P > .05). A concomitant increase in lactate and pyruvate production was noted with increasing concentrations of propionate. Succinate addition also increased lactate production by the ruminal epithelial cells. In contrast to propionate, succinate addition resulted in a decrease in beta-HBA formation from butyrate (P < .05). Butyrate metabolism by isolated ruminal epithelial cells is influenced by other VFA produced in the rumen, and propionate-induced stimulation of beta-HBA from butyrate does not seem to be mediated via succinate but rather is a result of a shift in the mitochondrial NADH/NAD status.

Acetates↗

The amino acid composition of protein feedstuffs before and after ruminal incubation and after subsequent passage through the intestines of dairy cows.

The amino (AA) profile of eight feedstuffs (two samples of fishmeal, soybean meal [SBM], formaldehyde-treated SBM, sopralin, cotton-seed meal [CSM], rapeseed meal [RSM], corn distillers grains [CDG], and corn gluten feed [CGF]) was determined before and after ruminal incubation for 8 and 12 h in three lactating Friesian cows using nylon bags. The profile of AA disappearing during intestinal passage was also measured by inserting the bags into the duodenum through T-piece cannulas after ruminal incubation and recovering them in the feces. The AA profile changed for all feedstuffs, except sopralin, following ruminal incubation. Changes were greater for the more degradable feedstuffs. The profile of AA disappearing during intestinal passage was generally similar to the profile after ruminal incubation, but some differences were found with feedstuffs that had low (< 84%) intestinal disappearance of AA (RSM, CDG, CGF). For the other feedstuffs, intestinal disappearance of AA was greater than 93%. The two fishmeal samples had the highest concentrations of methionine and lysine in their residues after ruminal incubation, whereas CDG and CGF had low lysine concentrations. Residues of these latter two and RSM were quite high in methionine concentration, whereas residues of SBM, sopralin, and CSM had the lowest concentrations. Corn distillers grains had 13% of its AA remaining after ruminal incubation followed by intestinal passage. These results show that feedstuffs vary in the proportion of their AA that escape ruminal degradation, in the profile of those AA, and in their intestinal digestibility. These factors should be considered in protein evaluation systems and in assessment of the protein quality of feedstuffs.

Amino Acids↗

Effects of laidlomycin propionate and monensin on the in vitro mixed ruminal microorganism fermentation.

The objective of this study was to compare the effects of laidlomycin propionate and monensin on the in vitro fermentation of ground corn, Trypticase, or alfalfa hay by mixed ruminal microorganisms. Ruminal fluid was collected from two steers fed 9.27 kg DM of a high-concentrate (62.2% ground corn and 17.4% cottonseed hulls) diet per day and composited. In the first study, no ionophore was included in the diet; the diet in the second study contained 11.1 g of laidlomycin propionate per ton of feed. The animals were allowed an adjustment period of 14 d for each dietary treatment before samples were collected. When ruminal fluid from unadapted animals was used, both monensin and laidlomycin propionate decreased (P<.05) CH4 concentration and the acetate:propionate ratio with ground corn and alfalfa hay. Monensin reduced (P<.05) in vitro dry matter disappearance of alfalfa and increased (P<.05) final pH in the ground corn and alfalfa hay fermentations. Both laidlomycin propionate and monensin decreased (P<.05) concentrations of acetate, propionate, isobutyrate, isovalerate, CH4, and NH3 in Trypticase fermentations. When ruminal fluid from adapted animals was used, both ionophores still reduced the concentrations of most fermentation products. However, there was generally less inhibition compared with fermentations inoculated with unadapted mixed ruminal microorganisms. In the presence of 5 mM maltose, mixed ruminal bacteria produced high concentrations (10 to 11 mM) of lactate, and addition of both ionophores to these fermentations was effective in reducing (P<.05) lactate production. In conclusion, laidlomycin propionate alters the mixed ruminal microorganism fermentation in a manner similar to monensin, but, at the concentrations used in this study, monensin seemed to be a more potent inhibitor.

Animal Feed↗

Changes in postprandial plasma and extracellular and ruminal fluid volumes in wethers fed or unfed for 72 hours.

Postprandial shifts in body water compartments might limit feed intake by ruminants, especially when an animal becomes partially dehydrated during transportation or other periods of water deprivation. This experiment was conducted to determine the effects of feed and water deprivation on postprandial changes in body water compartments in wethers. Hampshire wethers (n = 8; average BW 42 +/- 2 kg) were used in a crossover design. During each period, four wethers were limit-fed (540 g DM/d: FED) and four were deprived of feed and water for 72 h (DEPRIVED). Wethers were infused i.v. with Evans blue and sodium thiosulfate and intraruminally with Cr- or Co-EDTA, after which blood and ruminal samples were collected for the next 4 h. All wethers were then fed 540 g of feed DM, and infusions were repeated 30 min after feeding. Body water compartment volumes were determined with linear regression using plasma concentrations of Evans blue (plasma volume), and sodium thiosulfate (extracellular volume), and using ruminal fluid concentrations of Cr or Co. Feed and water deprivation decreased (P < .01) extracellular water space but did not affect plasma or ruminal water space. After feeding, extracellular water space decreased (P < .01) and ruminal volume increased (P < .05) in the FED and DEPRIVED wethers. Plasma pools of Na, K, and Mg were not affected by feeding in FED wethers but decreased (P < .05) in DEPRIVED wethers. The increase in ruminal fluid pools of Na, K, and Mg were greater (P < .05) in FED than in DEPRIVED wethers. These results indicate that abnormal water and electrolyte shifts may be factors partially responsible for the decreased feed intake by ruminants subjected to transportation or feed and water deprivation stress.

Animal Feed↗

Ruminal fermentation and duodenal flow following progressive inoculations of fauna-free wethers with major individual species of ciliate protozoa or total fauna.

Naturally fauna-free (FF) wethers, equipped with ruminal and duodenal cannulas, were used in two groups of eight (Group A) and seven (Group B) animals in six consecutive experimental periods, each lasting for 28 d. The objective was to measure ruminal fermentation traits, and flows of nonammonia nitrogen (NAN), total amino acid (TAA), and bacterial nitrogen (BN) from the stomach after inoculation with individual ciliate protozoa species in each period. The wethers in both groups were fed a diet based on corn silage, haylage, and soybean meal, and they remained FF during the first period. At the beginning of each other period, the wethers were progressively inoculated intraruminally with one individual major species of ruminal ciliate protozoa or total fauna (TF). Thus, Group A was progressively inoculated (+) with Dasytricha ruminantium (DS), Polyplastron multivesiculatum (PP), Isotricha intestinalis (IS), Entodinium caudatum (EN) and TF-type A. Also, Group B was progressively inoculated (+) with IS, DS, Epidinium ecaudatum (EP), Eudiplodinium maggi (EU), and EN. Duodenal digesta and ruminal fluid were collected and sampled in each period on d 26 and 28, respectively, and subjected to chemical analyses. A significantly higher (P < .05) pH (6.4) in ruminal fluid of the Group A wethers was obtained when each DS, DS+PP, DS-PP-IS+EN, and TF population was present in the rumen than when the wethers were FF (6.2). In the Group B wethers, pH (6.1) was lower (P < .05) for the population of IS-DS-EP+EU than for other populations (6.2 to 6.3). The concentration of total VFA in ruminal fluid was higher (P < .05) in the Group B wethers when IS, IS+DS, or IS-DS+EP populations were present in the rumen than when the wethers were FF. The flow of NAN, TAA, and BN from the stomach to the intestinal tract was generally lower for different protozoa populations than for the FF period. Largest decreases (P < .05) in the flow of NAN, TAA, and BN occurred when EN was added into the rumen of wethers in the A and B groups, which already contained populations of DS-PP+IS and IS-DS-EP+EU, respectively. Holotrich protozoa had very little effect on the protein metabolism in the rumen, but cellulolytic protozoa (PP, EP, and EU) and EN decreased the efficiency of protein utilization by the ruminant host.

Amino Acids↗

Influence of feed intake fluctuation and frequency of feeding on nutrient digestion, digesta kinetics, and ruminal fermentation profiles in limit-fed steers.

Nine crossbred beef steers (344 +/- 26 kg) fitted with ruminal cannulas were used in a randomized complete block design to evaluate the effects of feeding frequency and feed intake fluctuation on total tract digestion, digesta kinetics, and ruminal fermentation profiles in limit-fed steers. In Period 1, steers were allotted randomly to one of four dietary treatments: 1) feed offered once daily at 0800; 2) feed offered once daily at 0800 with a 10% fluctuation in day-to-day feed intake; 3) feed offered twice daily at 0800 and 1700; and 4) feed offered twice daily at 0800 and 1700 with a 10% fluctuation in a day-to-day feed intake. In Period 2, steers were reallocated across treatments. The 90% concentrate diet was fed at 90% of the ad-libitum consumption by each steer. Chromium-EDTA and Yb-labeled steam-flaked corn were intraruminally infused at 0800 on d 1 and 3 and Co-EDTA and Er-labeled steam-flaked corn were infused on d 2 and 4 of the 4-d collection period. Ruminal samples were collected at 0, 3, 6, 9, 12, 15, 18, and 24 h after the 0800 feeding, and total feces were collected for 4 d. Total tract digestibilities of OM, N, and starch were lowest (fluctuation x frequency, P < .05) when feed was offered twice daily with a 10% fluctuation in intake. Ruminal fluid volume and passage rate were not affected (P > .10) by feeding frequency or intake fluctuation. A frequency x fluctuation x sampling time interaction occurred (P < .01) for ruminal pH. Steers fed a constant amount of feed once daily had higher (P < .05) ruminal pH at 0, 3, 18, and 24 h than steers fed once daily with a 10% fluctuation in feed intake. Total VFA concentration was greater (P < .01) at 9 h after the 0800 feeding when feed was offered once vs twice daily. Feeding twice daily increased (P < .05) the molar proportion of acetate and decreased (P < .05) the molar proportion of propionate. Increasing feeding frequency resulted in a more stable ruminal environment; however, the increased acetate:propionate ratio with twice-daily feeding might result in lower efficiency of energy utilization by limit-fed steers.

Animal Feed↗

Supplemental soybean oil or corn for beef heifers grazing summer pasture: effects on forage intake, ruminal fermentation, and site and extent of digestion.

Nine Angus x Gelbvieh heifers (average BW = 347 +/- 2.8 kg) with ruminal and duodenal cannulas were used in a split-plot designed experiment to determine the effects of soybean oil or corn supplementation on intake, OM, NDF, and N digestibility. Beginning June 8, 1998, heifers continually grazed a 6.5-ha predominantly bromegrass pasture and received one of three treatments: no supplementation (Control); daily supplementation of cracked corn (Corn) at 0.345% of BW; or daily supplementation (0.3% of BW) of a supplement containing cracked corn, corn gluten meal, and soybean oil (12.5% of supplemental DM; Oil). Soybean oil replaced corn on a TDN basis and corn gluten meal was included to provide equal quantities of supplemental TDN and N. Three 23-d periods consisted of 14 d of adaptation followed by 9 d of sample collections. Treatment and sampling period effects were evaluated using orthogonal contrasts. Other than crude fat being greater (P = 0.01) for supplemented heifers, chemical and nutrient composition of masticate samples collected via ruminal evacuation did not differ (P = 0.23 to 0.56) among treatments. Masticate NDF and ADF increased quadratically (P < or = 0.003) and N decreased linearly (P = 0.0001) as the grazing season progressed. Supplementation did not influence (P = 0.37 to 0.83) forage OM intake, total and lower tract OM digestibility, ruminal and total tract NDF digestibility, or total ruminal VFA; however, supplemented heifers had lower ruminal molar proportions of acetate (P = 0.01), higher ruminal molar proportions of butyrate (P = 0.007), and greater quantities of OM digested in the rumen (P = 0.10) and total tract (P = 0.02). As the grazing season progressed, total tract OM and N and ruminal NH3 concentrations and NDF digestibility decreased quadratically (P < or = 0.04). Microbial N flow (P = 0.09) and efficiency (P = 0.04) and postruminal N disappearance (P = 0.02) were greater for Control heifers and declined linearly (P < or = 0.02) as the grazing season advanced. Depressed microbial N flow seemed to be more pronounced for Oil than for the Corn treatment. Although total digestible OM intake increased with supplementation, metabolizable protein supply was reduced in supplemented heifers. Therefore, feeding low levels of supplemental grain with or without soybean oil is an effective strategy to increase dietary energy for cattle grazing high-quality forages, but consideration should be given to reduced supply of metabolizable protein.

Animal Feed↗

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.

Amino Acids↗

Influence of supplementary fibrolytic enzymes on the fermentation of corn and grass silages by mixed ruminal microorganisms in vitro.

This study was done to determine the effectiveness of supplementary enzymes at increasing the fiber digestion by ruminal microorganisms and to assess whether enzyme activity limits the rate of fiber digestion in ruminal digesta. In vitro comparisons of enzyme activities in two feed enzyme preparations (A and B) with enzyme activities extracted from ruminal fluid indicated that the addition of fibrolytic enzymes at the application rates recommended by the manufacturers would not be expected to increase significantly glycanase and polysaccharidase activities in ruminal fluid. Preparations A and B both increased (P < 0.001) the rate of gas production from freeze-dried corn and grass silages in in vitro incubations with ruminal fluid, but only at concentrations much higher than recommended application rates. Autoclaved controls had little or no effect. Ultrafiltration of enzyme B indicated that most stimulation was due to components >100 kDa, which is consistent with the cause of the stimulation being enzyme activity. Fibrolytic enzymes from other sources were also able to stimulate gas production: increased rates of gas production were observed in seven out of eight combinations of "cellulase" and corn or grass silage (P < 0.05). The comparison of glycanase and polysaccharidase activities with gas-stimulatory activity in the different enzyme preparations indicated that the highest correlation was between increased gas production and enzyme activity against microgranular cellulose (P < 0.05). In a wider range of fibrolytic enzyme preparations, those with endo-(beta-1,4)- or exo-(beta-1,4)-xylanase activity equal to that of preparation A did not produce similar increased rates of fermentation of corn silage when glucanase activity was low (P > 0.05). In contrast, preparations with glucanase activity similar to enzyme A gave at least as great (P < 0.05) an improvement in gas production than enzyme A, irrespective of xylanase activity. It was concluded that enzyme activity, probably a type of endo-(beta-1,4)-glucanase activity, limits the rate of fermentation of corn and grass silage in the rumen. Enzyme supplements of the type used in these experiments are unlikely to possess sufficient activity to overcome this limitation by direct application to ruminal digesta, implying that treatment of the ration prefeeding will be key to harnessing the potential of exogenous fibrolytic enzymes in ruminant nutrition.

Animal Feed↗

Apparent ruminal degradation and rumen escape of soluble nitrogen fractions in grass and grass silage administered intraruminally to lactating dairy cows.

The main objective of this study was to investigate in vivo ruminal degradation and rumen escape of soluble N fractions in grass and grass silage. Soluble protein and long-chain peptides (PLP), small peptides (SP) and free AA (FAA) were obtained from fresh grass and grass silages fertilized with different levels of N. Soluble extracts from the forages were pulse dosed into the rumen of three cannulated lactating dairy cows, and a simple or complex model was used to examine the kinetics of the soluble N fractions in the rumen. When soluble extracts from silage were investigated, pulse dosages of total nonammonia N (NAN) were 21, 27, and 32 g, while for fresh grass only dosages of 20 g were ruminally administered. In the silage extracts, mean proportions of PLP-N, SP-N, and FAA-N in the NAN were 30, 52, and 18%, respectively, whereas in the fresh grass the corresponding values were 67, 20, and 13%. From silage extracts, all three soluble N fractions showed a linear decrease (P < 0.05) in degradation rate and an increase (P < 0.05) in ruminal escape with increasing dosage. In silage, mean degradation rates, parameterized from the complex model, were 230, 214, and 334%/h for PLP-N, SP-N, and FAA-N, respectively, and the ruminal escape was highest (P < 0.05) for SP-N (11.2% of dose) and lowest (P < 0.05) for FAA-N (5.0% of dose). No differences in degradation rate and ruminal escape between fresh grass and silage were observed. However, the proportion of N dose converted to ammonia was only 24% in the fresh grass, whereas for the silages a mean value of 76% was found. From this study, it is concluded that a significant amount of dietary soluble N escapes ruminal degradation, and thus contributes to the intestinal AA supply. Moreover, if the main aim is to study degradation kinetics of individual N fractions, a complex model should be used in the evaluation. This model can also be used to study ruminal synchronization of N and energy for microbial growth.

Animal Feed↗

Net absorption and utilization of nitrogenous compounds across ruminal, intestinal, and hepatic tissues of growing beef steers fed dry-rolled or steam-flaked sorghum grain.

Our objectives were to determine effects of grain processing on splanchnic (gut tissues and liver) N metabolism and whole-body N balance by growing steers and to ascertain the relative contributions of ruminal and intestinal tissues to net absorption and utilization of N-containing nutrients. Seven beef steers (348 kg initial BW), surgically implanted with appropriate catheters, were fed diets containing 77% steam-flaked (SF) or dry-rolled (DR) sorghum grain. Blood flows and net output or uptake of ammonia N, urea N, and alpha-amino N (estimate of amino acids) were measured across portal-drained viscera (PDV or gut tissues) and intestinal, ruminal, hepatic, and splanchnic tissues (PDV + hepatic). The experimental design was a crossover between DR and SF diets, with six samplings of blood at 2-h intervals on 2 d for each steer. Nitrogen intake (139 +/- 3 g/d), output in urine (43 +/- 2 g/d), and retention (40 +/- 3 g/d) were similar for both processing treatments. When steers were fed SF sorghum compared to DR sorghum, N retention as a percentage of N intake was numerically greater (P < 0.12), output of fecal N was numerically lower (P < 0.13), and urinary urea N was lower (P < 0.04). For SF vs DR, net uptake of alpha-amino N by liver was higher (P < 0.04; 20 vs 9 g/d) and was numerically lower (P < 0.16) for ruminal tissues (15 vs 33 g/d). Feeding steers SF compared to DR tended to increase net transfer (cycling) of blood urea N to PDV (57 vs 41 g/d; P < 0.07), increased cycling to intestinal tissues (15 vs 6 g/d; P < 0.05), and numerically increased transfer to ruminal tissues (42 vs 32 g/d; P < 0.12) but did not alter other net output or uptake of N across splanchnic tissues. Total urea N transfer (blood + saliva) was similar for both treatments. Net uptake of alpha-amino N by ruminal tissues was about 30% of the net amount of alpha-amino N absorbed across the intestinal tissues. In summary, most of the blood urea N cycled from the liver to gut tissues was transferred to ruminal tissues for potential microbial protein synthesis, and the net ruminal utilization of alpha-amino N was about 30% of that absorbed from intestinal tissues. Feeding growing steers SF compared to DR sorghum diets numerically increased whole-body N retention (percentage of N intake) by about 15% and tended to increase transfer of blood urea N to the gut by about 40%, which could increase the supply of high-quality microbial protein for absorption.

Absorption↗