Amino acid composition of rumen bacteria and protozoa in cattle

Because microbial crude protein (MCP) constitutes more than 50% of the protein digested in cattle, its AA composition is needed to adequately estimate AA supply. Our objective was to update the AA contributions of the rumen microbial AA flowing to the duodenum using only studies from cattle, differentiating between fluid-associated bacteria (FAB), particle-associated bacteria (PAB), and protozoa, based on published literature (53, 16, and 18 treatment means were used for each type of microorganism, respectively). In addition, Cys and Met reported concentrations were retained only when an adequate protection of the sulfur groups was performed before the acid hydrolysis. The total AA (or true protein) fraction represented 82.4% of CP in bacteria. For 10 AA, including 4 essential AA, the AA composition differed between protozoa and bacteria. The most noticeable differences were a 45% lower Lys concentration and 40% higher Ala concentration in bacteria than in protozoa. Differences between FAB and PAB were less pronounced than differences between bacteria and protozoa. Assuming 33% FAB, 50% PAB, and 17% of protozoa in MCP duodenal flow, the updated concentrations of AA would decrease supply estimates of Met, Thr, and Val originating from MCP and increase those of Lys and Phe by 5 to 10% compared with those calculated using the FAB composition reported previously. Therefore, inclusion of the contribution of PAB and protozoa to the duodenal MCP flow is needed to adequately estimate AA supply from microbial origin when a factorial method is used to estimate duodenal AA flow. Furthermore, acknowledging the fact that hydrolysis of 1 kg of true microbial protein yields 1.16 kg of free AA substantially increases the estimates of AA supply from MCP.     

Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture

Eight continuous culture fermentors inoculated with ruminal liquor from heifers fed a 50:50 alfalfa hay:concentrate diet (17.6% crude protein, 28.0% neutral detergent fiber) were used in 3 replicated periods to study the effects of cinnamaldehyde (CIN) and garlic oil (GAR) on rumen microbial fermentation. Treatments were no additive (negative control); 1.25 mg/L (MON) and 12.5 mg/L (MON10) of the ionophore antibiotic monensin (positive control); 31.2 mg/L CIN (CIN) and 312 mg/L (CIN10) of CIN; and 31.2 mg/L GAR (GAR) and 312 mg/L (GAR10) of GAR (Allium sativa). The MON10 caused expected changes in microbial fermentation patterns (a decrease in fiber digestion, ammonia N concentration, and proportions of acetate and butyrate; an increase in the proportion of propionate; and a trend to increase small peptide plus AA N concentration). The CIN decreased the proportion of acetate and branch-chained volatile fatty acids (VFA) and increased the proportion of propionate; CIN10 decreased the proportion of acetate and increased the proportion of butyrate compared with the control. The GAR10 increased the proportion of propionate and butyrate and decreased the proportion of acetate and branch-chained VFA compared with the control. The GAR10 also increased the small peptide plus amino acid N concentration, although no effects were observed on large peptides or ammonia N concentrations. The CIN and GAR10 resulted in similar effects as monensin, with the exception of the effects on the molar proportion of butyrate, which suggests that they might have a different mode of action in affecting in vitro microbial fermentation.     

Essential oil and monensin affect ruminal fermentation and the protozoal population in continuous culture

The interaction of monensin and essential oil was hypothesized to suppress protozoa and methane production while maintaining normal rumen function. The objective of this study was to determine the effects of feeding monensin (MON) and CinnaGar (CIN, a commercial blend of cinnamaldehyde and garlic oil; Provimi North America, Brookville, OH) on ruminal fermentation characteristics. Continuous culture fermentors (n = 4) were maintained in 4 experimental periods in a 4 × 4 Latin square design. Four dietary treatments were arranged in a 2 × 2 factorial: (1) control diet, 37 g/d of dry matter (40 g/d at ～92.5% dry matter) of a 50:50 forage:concentrate diet containing no additive; (2) MON at 11 g/909 kg of dry matter; (3) CIN at 0.0043% of dry matter; and (4) a combination of MON and CIN at the levels in (2) and (3). Treatment had no effects on protozoal populations, concentration of NH₃N, total N flow of effluent, production of total volatile fatty acids, or flows of conjugated linoleic acid and total C18 fatty acids. The MON decreased acetate:propionate ratio and biohydrogenation of both total C18 and 18:1 cis-9 but increased protozoal generation time, concentration of peptide, and flow of 18:1 trans-11. The MON tended to decrease protozoal counts in effluent and flow of 18:0 but tended to increase propionate production. The CIN decreased true organic matter digestibility and protozoal N flow of effluent but increased nonammonia, nonmicrobial N flow. The CIN tended to decrease protozoal counts, microbial N flow, and neutral detergent fiber digestibility but tended to increase biohydrogenation of total C18, 18:2, and 18:3. The CIN tended to increase isovalerate production. The MON and CIN tended to interact for increased methane production and bacterial N flow. A second experiment was conducted to determine the effects of MON and CIN on protozoal nitrogen and cell volume in vitro. Four treatments included (1) control (feed only), (2) feed + 0.0043% dry matter CIN, (3) feed + 2.82 μM MON, and (4) feed + CIN + MON at the same levels as in (2) and (3). With no interactions, MON addition decreased percentage of protozoa that were motile and tended to decrease cell volume at 6 h. The CIN did not affect cell count or other indicators of motility or volume at either 3 or 6 h. Under the conditions of our study, we did not detect an additive response for MON and CIN to decrease protozoal counts or methane production. A 3-dimensional method is suggested to better estimate protozoal cell volume.     

Effects of Cordyceps militaris on the growth of rumen microorganisms and in vitro rumen fermentation with respect to methane emissions

This experiment was designed to investigate the effects of different concentrations (0.00, 0.10, 0.15, 0.20, 0.25, and 0.30 g/L) of dried Cordyceps militaris mushroom on in vitro anaerobic ruminal microbe fermentation and methane production using soluble starch as a substrate. Ruminal fluids were collected from Korean native cattle, mixed with phosphate buffer (1:2), and incubated anaerobically at 38°C for 3, 6, 9, 12, 24, 36, 48, and 72 h. The addition of C. militaris significantly increased total volatile fatty acid and total gas production. The molar proportion of acetate was decreased and that of propionate was increased, with a corresponding decrease in the acetate:propionate ratio. As the concentration of C. militaris increased from 0.10 to 0.30 g/L, methane and hydrogen production decreased. The decrease in methane accumulation relative to the control was 14.1, 22.0, 24.9, 39.7, and 40.9% for the 0.10, 0.15, 0.20, 0.25, and 0.30 g/L treatments, respectively. Ammonia-N concentration and numbers of live protozoa decreased linearly with increasing concentrations of C. militaris. The pH of the medium significantly decreased at the highest level of C. militaris compared with the control. In conclusion, C. militaris stimulated mixed ruminal microorganism fermentation and inhibited methane production in vitro. Therefore, C. militaris could be developed as a novel compound for antimethanogenesis.     

Effects of patterns of suboptimal pH on rumen fermentation in a dual-flow continuous culture system

Low ruminal pH affects rumen fermentation, with the effects being larger as the time at suboptimal pH increases. Eight 1,325-mL dual-flow continuous culture fermenters were used to examine the hypothesis that the negative effects of a single cycle of 12 h (experiment 1) or 8 h (experiment 2) at pH 5.5 can be reduced by splitting it into several cycles. Temperature (39°C), diet (97 g/d of a 60:40 forage:concentrate diet), and solid (5%/h) and liquid (10%/h) dilution rates were kept constant. In experiment 1, treatments were a constant pH 6.4 (H); 1 cycle of 12 h at pH 5.5 (L12); 2 cycles of 6 h at pH 5.5; and 3 cycles of 4 h at pH 5.5. In experiment 2, treatments were a constant pH 6.4 (H); 1 cycle of 4 h at pH 5.5 (L4); 1 cycle of 8 h at pH 5.5 (L8); or 2 cycles of 4 h at pH 5.5. During the rest of the day, pH was maintained at 6.4. Each experiment consisted of 2 replicated periods of 8 d (5 d for adaptation and 3 d for sampling). Within period, treatments were randomly assigned to fermenters. Data were analyzed as a randomized complete block using PROC MIXED of SAS and differences declared at P < 0.05 using the Tukey's test. In experiment 1, L12 reduced neutral detergent fiber (NDF) digestion, acetate proportion, and the acetate:propionate ratio, increased propionate proportion, and tended to reduce ammonia N concentration, compared with H, but had no effect on the flow of dietary or microbial N, crude protein degradation, efficiency of microbial protein synthesis, or the flow of total, essential, and individual amino acids. Dividing the 12 h at suboptimal pH into 2 or 3 cycles reduced true organic matter (OM) degradation compared with H, and did not alleviate the negative effects on NDF digestion and volatile fatty acid profile observed in L12. In experiment 2, L4 tended to reduce true OM digestion, ammonia N concentration, and bacterial N flow, reduced CP degradation, and increased dietary N flow. Treatment L8 reduced OM and NDF digestion, and ammonia N concentration, compared with H. Treatments L4 and L8 also reduced acetate proportion and the acetate:propionate ratio, and increased propionate proportion and the flow of total, essential, and most individual amino acids, but had no effect on efficiency of microbial protein synthesis compared with the H treatment. When the 8 h at suboptimal pH was divided into 2 cycles of 4 h the effects were not different from L8. Results suggest that the effects of low pH are dependent on the total amount of time that pH is suboptimal and are not reduced by splitting it into various cycles.     

Effects of time at suboptimal pH on rumen fermentation in a dual-flow continuous culture system

Ruminal pH varies considerably during the day, achieving values below 6.0 when cows consume large amounts of concentrates. Low ruminal pH has negative effects on ruminal fermentation. However, previous studies have indicated that rumen bacteria may resist short periods of low ruminal pH, and it is not clear how long this period may be before rumen microbial fermentation is negatively affected. Seven dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods with the same diet (97 g of dry matter/d of a 60:40 forage-to-concentrate diet, 18.3% crude protein, 35.9% neutral detergent fiber), temperature (39°C), and solid (5%/h) and liquid (10%/h) dilution rates to study the effects of increasing time at suboptimal pH on rumen microbial fermentation and nutrient flow. Treatments were a constant pH of 6.4 and 6 different intervals of time during the day (4, 8, 12, 16, 20, 24 h) at suboptimal pH (5.5), with the rest of the day being at pH 6.4. Polynomial equations were derived using the Mixed procedure of SAS, and linear, quadratic and cubic terms were left in the equation if P < 0.10. True organic matter digestion decreased with increasing time at suboptimal pH and was best described by a cubic regression (TOMD = 58.5 − 2.15x + 0.16x² −0.0037x³; R² = 0.74). Digestion of NDF (DNDF = 55.1 − 1.00x; R² = 0.75) and digestion of ADF (DADF = 56.2 − 1.33x; R² = 0.78) decreased linearly with increasing time at suboptimal pH. Total VFA had a cubic response (VFA = 112.7 − 2.09x + 0.17x² − 0.0054x³; R² = 0.82). The proportion of acetate decreased linearly (acetate = 58.7 − 0.61x; R² = 0.79). The propionate proportion increased (propionate = 17.6 + 2.09 × −0.044x²; R² = 0.85) and branched-chain VFA decreased (BCVFA = 4.45 −0.51x + 0.014x²; R² = 0.75) quadratically. The ammonia N concentration (NH₃-N = 5.85 − 0.13x; R² = 0.46) and flow (NH₃-N flow = 0.18 − 0.0039x; R² = 0.43) decreased linearly as the time at suboptimal pH increased. Crude protein degradation (CPd = 41.9 − 1.60x + 0.060x²; R² = 0.71), efficiency of microbial protein synthesis (EMPS = 26.6 − 0.33x + 0.021x²; R² = 0.77), microbial N flow (MN flow = 1.38 − 0.036x + 0.0015x²; R² = 0.77), and dietary N flow (DN flow = 1.49 + 0.041x − 0.0015x²; R² = 0.65) had a quadratic response. The flow of essential, nonessential, and most individual AA increased linearly with increasing time at suboptimal pH. The effects of pH on rumen fermentation appear to start as soon as pH drops to suboptimal pH.     

Effects of BioChlor and Fermenten on microbial protein synthesis in continuous culture fermenters

Meta analysis models were constructed from a data-set of 15 continuous culture fermenter trials and 118 observations on studies with either BioChlor (n = 23 observations) or Fermenten (n = 95) included at 10 and 3%, respectively, of dietary dry matter (DM) to evaluate effects of the ingredients BioChlor and Fermenten (B/F) on rumen function. Digestibility of crude protein was significantly increased by 11% with B/F treatment. This was reflected in significant increases in digestibility of DM and organic matter (OM) by 3.6 and 7.9%, respectively. Increased amounts of sugar in the diet in the presence of B/F tended to reduce digestibility of non-structural carbohydrates (NSC); however, the net effect on NSC digestion was small. There was no effect of treatment on most individual volatile fatty acids (VFA) or total VFA production. Propionate production, however, was significantly reduced in treated fermenters. The main effect of B/F as well as of starch and soluble fiber when combined with the treatment was to increase propionate production; however, the interaction between B/F treatment and sugar decreased propionate production markedly, resulting in a net decrease. The acetate-to-propionate ratio increased by 6% with B/F, largely as a result of the decrease in propionate. Production of nonammonia nitrogen was 1% less in B/F-treated fermenters, and interactions between treatment and starch, sugar, or soluble fiber were significant. Treated fermenters produced 15.7% more microbial nitrogen, in association with a significant 37% increase in rumen protein digestion. Interactions between treatment and starch, soluble fiber, or sugar influenced these results. The interaction of B/F and sugar resulted in a decrease in undegradable protein N and an increase in microbial nitrogen production. Ammonia nitrogen concentrations were increased by 24.6% in treated fermenters. Efficiency of microbial nitrogen production from DM, OM, or carbohydrate was significantly increased by B/F. Sugar content increased efficiency of microbial protein production/kg of OM digested or carbohydrate digested in the presence of treatment by >10 times the increase that was attributable to the interaction of treatment with starch. Treatment with B/F reduced moles of VFA produced/kg of microbial nitrogen produced by 16%. This effect was also substantially influenced by interactions between B/F and sugar. If the fermenter results are representative of those in vivo, milk production responses to treatment with B/F will depend on amounts of starch, soluble fiber, and, particularly, sugar in diets. Milk production responses will also depend on the quality of protein in the diet and the comparative benefit that increased flux of microbial nitrogen provides. Increased digestibility of OM should allow additional ruminant production benefits.     

Effects of extracts of spices on rumen methanogenesis,enzyme activities and fermentation of feeds in vitro

BACKGROUND: An experiment was conducted to study the effects of boiling water, methanol and ethanol extracts (0, 0.25 and 0.50 mL) of seeds of Foeniculum vulgare (fennel), flower buds of Syzygium aromaticum (clove), bulbs of Allium sativum (garlic), bulbs of Allium cepa (onion) and roots of Zingiber officinalis (ginger) on rumen methanogenesis, fibrolytic enzyme activities and fermentation characteristics in vitro. RESULTS: Ethanol and methanol extracts of fennel, clove and garlic at 0.50 mL and clove at 0.25 mL inhibited (P < 0.05) methane production. Carboxymethylcellulase activity was reduced (P < 0.05) by ethanol and methanol extracts (0.50 mL) of fennel and clove (0.25 and 0.50 mL). The extracts of clove reduced (0.25 and 0.50 mL) xylanase and acetylesterase activities, and the fennel extract (0.50 mL) reduced (P < 0.05) xylanase activity. However, the extracts of garlic (0.50 mL) increased (P < 0.05) acetylesterase activity. Concentrations of volatile fatty acids were reduced (P < 0.05) by the extracts of garlic and onion. The extracts of garlic caused a decrease (P < 0.05) in acetate:propionate ratio (A:P) at 0.50 mL, whereas A:P was increased (P < 0.05) by the inclusion of 0.50 mL extracts of clove. Methanol and ethanol extracts of clove decreased (P < 0.05) in vitro organic matter degradability. Extracts (0.50 mL) of clove decreased (P < 0.05) the numbers of total protozoa, small entodiniomorphs and holotrichs, whereas extracts of onion, ginger and garlic enhanced (P < 0.05) protozoal numbers (both entodiniomorphs and holotrichs). CONCLUSION: Ethanol and methanol extracts of fennel and garlic have potential to inhibit rumen methanogenesis without adversely affecting rumen fermentation. Copyright © 2009 Society of Chemical Industry     

Effect of humic acid on fermentation and ciliate protozoan population in rumen fluid of sheep in vitro

BACKGROUND: Humic acid (HA) as a product of decomposition of animal and plant tissue is used in animal production as a feed supplement, antimicrobial product and growth stimulator. The objective of the present in vitro study was to investigate the effects of dietary humic acid as a commercial Humacid 60 Basic (H60B) feed additive preparation on rumen fermentation and the ciliate protozoan population in the rumen fluid of sheep using a high fibre (HF) and high concentrate (HC) diet in batch cultures and artificial rumen (RUSITEC). RESULTS: Production of total volatile fatty acids (VFAs) was significantly decreased (P < 0.001) for batch cultures by the HF‐H60B diet. The HF‐H60B diet decreased ammonia N in RUSITEC (P < 0.001). An increase in the population of Enoploplastron triloricatum, Isotricha spp. and Ophryoscolex c. tricoronatus with the HF‐H60B diet and Diploplastron affine with the HC‐H60B was observed. The H60B did not affect the total ciliate population and Entodinium spp. CONCLUSION: It can be concluded that dietary humic acid preparations are not effective as dietary antiprotozoal agents. Humic acid might enhance microbial growth and energy efficiency in doses up to 10 g kg^?1 DM of diet. Copyright © 2009 Society of Chemical Industry     

Investigation of ammonium lactate supplementation on fermentation end products and bacterial assimilation of nitrogen in dual-flow continuous culture

Supplements investigated throughout the present study are produced by fermenting lactose that is present in whey to lactate, yielding products differing in ammonium relative to lactate concentrations and in physical form (liquid or dry). Trials 1 and 2 investigated Lacto-Whey (LW; Fermented Nutrition Corp., Luxemburg, WI) and GlucoBoost (GB; Fermented Nutrition Corp.), respectively, using dual-flow continuous culture systems (n = 4), each with a 4 × 4 Latin square design. A greater proportion of nonprotein nitrogen was present in GB than in LW. In trial 1, the treatment with LW was isonitrogenously dosed against soybean meal (SBM) as a control (no LW) and factorialized with either a wheat- or corn-based concentrate (55% inclusion rate, dry matter basis). We hypothesized that LW would increase propionate production and that the combination of +LW with wheat would increase bacterial assimilation of NH₃-N into cellular N. No differences were observed for total volatile fatty acid (VFA) production per day. However, treatment × time interactions revealed that +LW increased lactate concentration at 0, 0.5, and 1 h and tended to increase molar percentage of propionate at 1 and 1.5 h postfeeding, documenting the immediate availability of lactate converted to propionate in the +LW treatments. The main effect of corn increased the proportion of bacterial N derived from NH₃-N. Trial 2 was designed to investigate GB; isonitrogenous treatments included an SBM control, crystal GB, liquid GB (LGB), and LGB with yeast culture, which were dosed twice daily. We hypothesized that GB would increase propionate production and bacterial assimilation of NH₃-N; the combination of LGB and yeast culture was expected to have a positive additive effect, yielding the greatest VFA production and bacterial NH₃-N assimilation. No differences were observed for total VFA production; however, LGB decreased molar percentage of acetate and increased propionate and butyrate molar percentages. There were no differences in non-NH₃-N flow or microbial N flow. Under the conditions of our studies, lactate in LW and GB was fermented extensively to propionate, and microbial protein synthesis in these treatments was comparable with that in SBM controls.     

Effects of peripartal yeast culture supplementation on lactation performance,blood biomarkers,rumen fermentation,and rumen bacteria species in dairy cows

Feeding yeast culture fermentation products has been associated with improved feed intake and milk yield in transition dairy cows. These improvements in performance have been further described in terms of rumen characteristics, metabolic profile, and immune response. The objective of this study was to evaluate the effects of a commercial yeast culture product (YC; Culture Classic HD, Phibro Animal Health) on performance, blood biomarkers, rumen fermentation, and rumen bacterial population in dairy cows from −30 to 50 d in milk (DIM). Forty Holstein dairy cows were enrolled in a randomized complete block design from −30 to 50 DIM and blocked according to expected calving day, parity, previous milk yield, and genetic merit. At −30 DIM, cows were assigned to either a basal diet plus 114 g/d of ground corn (control; n = 20) or a basal diet plus 100 g/d of ground corn and 14 g/d of YC (n = 20), fed as a top-dress. Cows received the same close-up diet from 30 d prepartum until calving [1.39 Mcal/kg of dry matter (DM) and 12.3% crude protein (CP)] and lactation diet from calving to 50 DIM (1.60 Mcal/kg of DM and 15.6% CP). Blood samples and rumen fluid were collected at various time points from −30 to 50 d relative to calving. Cows fed YC compared with control showed a trend for increased energy-corrected milk (+3.2 kg/d). Lower somatic cell counts were observed in YC cows than in control. We detected a treatment × time interaction in nonesterified fatty acids (NEFA) that could be attributed to a trend for greater NEFA in YC cows than control at 7 DIM, followed by lower NEFA in YC cows than control at 14 and 30 DIM. In the rumen, YC contributed to mild changes in rumen fermentation, mainly increasing postpartal valerate while decreasing prepartal isovalerate. This was accompanied by alterations in rumen microbiota, including a greater abundance of cellulolytic (Fibrobacter succinogenes) and lactate-utilizing bacteria (Megasphaera elsdenii). These results describe the potential benefits of supplementing yeast culture during the late pregnancy through early lactation, at least in terms of rumen environment and performance.     

Fermentation characteristics of corn‐, triticale‐, and wheat‐based dried distillers' grains with solubles in barley‐based diets determined using continuous and batch culture systems

BACKGROUND: Dried distillers' grains with solubles (DDGS) are becoming increasingly available and popular as an alternative livestock feed. This study used continuous and batch culture techniques to compare the in vitro fermentation characteristics of diets containing corn‐, triticale‐ or wheat‐based DDGS at 200 g kg^?1 diet dry matter (DM) against a barley grain‐based control diet. RESULTS: In continuous fermentation of wheat DDGS diet, total volatile fatty acid (VFA) concentration was decreased by 15.7% across sampling times and NH₃ concentration was quadrupled compared with control, whereas fermentations of corn‐ and triticale‐DDGS diets were similar to the barley‐based control. In batch cultures, corn DDGS differed from control only in increased culture pH. Compared with control diet, triticale DDGS yielded lower total gas production (140.94 versus 148.78 mL g^?1 DM) and in vitro DM digestibility (IVDMD; 0.509 versus 0.535). Wheat DDGS diet yielded decreased total gas production (123.49 mL g^?1 DM) and IVDMD (0.468), as well as total VFA production (105.84 versus 134.20 mmol L^?1) and substantially increased ammonia concentration (151.61 versus 58.34 mg L^?1) and acetate:propionate ratio (2.94 versus 1.11). CONCLUSION: Corn‐ and triticale‐ DDGS diets exhibited fermentation characteristics similar to the barley based control diet, consistent with in vivo findings that these diets yielded no adverse effects on production. In vitro ruminal fermentation of wheat DDGS diet differed significantly from control in several aspects including 2.6 to 6X higher ammonia concentrations. Copyright © 2010 Crown in the right of Canada. Published by John Wiley & Sons, Ltd     

Metabolism of small peptides in rumen fluid. Accumulation of intermediates during hydrolysis of alanine oligomers,and comparison of peptidolytic activities of bacteria and protozoa

Oligopeptides of L -alanine up to ala₅, were incubated in vitro in either strained rumen fluid or suspensions of mixed rumen bacteria. The disappearance of substrates and formation of products were measured for 40 min, by which time ala₃, ala₄ and ala₅ were almost totally hydrolysed. Ala₂ was more slowly hydrolysed, and accumulated in incubations with the other peptides. The pattern of formation of ala₂ but not ala₃ from ala₄, and ala₃ and ala₂ but not ala₄ from ala₅, suggested that the peptides were being hydrolysed by a dipeptidyl peptidase mechanism. Three different sheep receiving different diets gave similar results. Neither substrates nor products appeared to be accumulated intracellularly, except for ala₂, and then only if protozoa were present. Protozoa were more active than bacteria in ala₂ hydrolysis, whereas bacteria had greater activities with higher homologues. Similar preferences were observed with glycine peptides, although unlike alanine peptides gly₃ and gly₅ were more slowly degraded than the dimer. These experiments suggest that protozoa are of importance in the accumulation and hydrolysis of dipeptides, whereas bacteria are responsible for the breakdown of larger molecules by a dipeptidyl peptidase mechanism that does not appear to involve accumulation within the cell.     

Mode of action of yeast culture (YEA-SACC 1026) for stimulation of rumen fermentation in buffalo calves

The effect of daily supplementation of 5 g Saccharomyces cerevisiae yeast culture (YC, YEA-SACC 1026), 30 g NaHCO₃, supernatant from 5 g YC (YCS), 5 g autoclaved YC (YCH) or 5 g γ-irradiated YC (YCR) to the diet of buffalo calves on rumen microbial populations and fermentation pattern was examined. Addition of 30 g NaHCO₃ increased the rumen pH to the level observed with YC group. The pH and the concentrations of total, total viable and cellulolytic bacteria and total volatile fatty acids (VFA) were significantly higher while that of lactic acid, hexose-unit oligosaccharides and NH₃-N were significantly lower in the rumen fluid of YC compared with the control group. The effect of NaHCO₃ was 39·5 and 59·5% in decreasing the concentrations of lactic acid and hexose-unit oligosaccharides, 48·1, 47·2 and 45·5% in increasing the numbers of total, total viable and cellulolytic bacteria, 50·0 and 58·1% in increasing the concentrations of total VFA and protein and 51·3% in decreasing the concentration of NH₃-N of YC. The corresponding values for YCR addition in the diet were 38·6, 45·7, 48·5, 44·4, 51·5, 39·1, 48·1 and 46·5%. The effect of YCS and YCH was only marginal, but conspicuous up to 2 h after feeding, in changing the above rumen variables when compared with the YC group. The results indicated that contribution of increase in pH in changing the rumen variables was approximately 50% of YC and almost all the stimulatory activity was associated with live yeast cells. Autoclaving of YC destroyed almost all and γ-irradiation of YC retained about 50% of stimulatory activity of YC. The effect of YC on rumen fermentation, which was maximum up to 2 to 4 h after feeding, decreased with time. © 1998 SCI.     

Conditions stimulating neutral detergent fiber degradation by dosing branched-chain volatile fatty acids. II: Relation with solid passage rate and pH on neutral detergent fiber degradation and microbial function in continuous culture

To support improving genetic potential for increased milk production, intake of digestible carbohydrate must also increase to provide digestible energy and microbial protein synthesis. We hypothesized that the provision of exogenous branched-chain volatile fatty acids (BCVFA) would improve both neutral detergent fiber (NDF) degradability and efficiency of microbial protein synthesis. However, BCVFA should be more beneficial with increasing efficiency of bacterial protein synthesis associated with increasing passage rate (k_p). We also hypothesized that decreasing pH would increase the need for isobutyrate over 2-methylbutyrate. To study these effects independent from other sources of variation in vivo, we evaluated continuous cultures without (control) versus with BCVFA (0 vs. 2 mmol/d each of isobutyrate, isovalerate, and 2-methylbutyrate), low versus high k_p of the particulate phase (2.5 vs. 5.0%/h), and high versus low pH (ranging from 6.3 to 6.8 diurnally vs. 5.7 to 6.2) in a 2 × 2 × 2 factorial arrangement of treatments. Diets were 50% forage pellets and 50% grain pellets administered twice daily. Without an interaction, NDF degradability tended to increase from 29.7 to 35.0% for main effects of control compared with BCVFA treatments. Provision of BCVFA increased methanogenesis, presumably resulting from improved NDF degradability. Decreasing pH decreased methane production. Total volatile fatty acid (VFA) and acetate production were decreased with increasing k_p, even though true organic matter degradability and bacterial nitrogen flow were not affected by treatments. Decreasing pH decreased acetate but increased propionate and valerate production, probably resulting from a shift in bacterial taxa and associated VFA stoichiometry. Decreasing pH decreased isobutyrate and isovalerate production while increasing 2-methylbutyrate production on a net basis (subtracting doses). Supplementing BCVFA improved NDF degradability in continuous cultures administered moderate (15.4%) crude protein diets (excluding urea in buffer) without major interactions with culture pH and k_p.     

Effect of supplemental yeast culture and dietary starch content on rumen fermentation and digestion in dairy cows

The objectives of this experiment were to evaluate the effect of feeding a culture of Saccharomyces cerevisiae on rumen metabolism and digestibility when cows are fed diets varying in starch content. Four lactating Holstein cows were assigned to a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Treatments were low starch (LS; 23% of diet DM) and no yeast culture (YC; LS-control), LS and 15 g of YC/d (LS-YC), high starch (HS; 29% of diet DM) and no YC (HS-control), and HS and 15 g of YC/d (HS-YC). Periods lasted 28 d, with the last 9 d for data collection. Days 20 to 24 were used to determine production, nutrient flow, and digestibility. On d 25, 3 kg of corn grain DM was placed in the rumen 1 h before the morning feeding, and yields of milk and milk components were measured after the challenge. Blood was sampled −1, 3, 7, and 11 h relative to the morning feeding on d 24 and 25. Rumen pH was measured continuously on d 24 and 25. Rumen papillae were collected on d 24 and 28 to quantify mRNA expression of select genes. Supplementing YC increased yields of milk (26.3 vs. 29.6 kg/d), energy-corrected milk (ECM; 26.5 vs. 30.3 kg/d), fat (0.94 vs. 1.08 kg/d), true protein (0.84 vs. 0.96 kg/d), and ECM/dry matter intake (1.15 vs. 1.30) compared with the control but did not affect dry matter intake (22.6 vs. 22.9 kg/d). Cows fed HS had increased milk true protein percentage (3.18 vs. 3.31%) and yield (0.87 vs. 0.94 kg/d) compared with cows fed LS. Feeding HS-YC increased the proportion of dietary N incorporated into milk true protein from 24.9% in the other 3 treatments to 29.6%. Feeding HS increased the concentration of propionate (21.7 vs. 32.3 mM) and reduced that of NH₃-N (8.3 vs. 6.7 mg/dL) in rumen fluid compared with the control, and combining HS with YC in HS-YC tended to increase microbial N synthesis compared with LS-YC (275 vs. 322 g/d). Supplementing YC to cows fed HS reduced plasma haptoglobin and rumen lactate concentrations, increased mean rumen pH, reduced the time with pH <6.0, and prevented the decrease in rumen neutral detergent fiber digestion caused by HS. Cows fed HS had less total-tract digestion of organic matter (73.9 vs. 72.4%) because of reduced acid detergent fiber (57.6 vs. 51.7%) and neutral detergent fiber (60.9 vs. 56.7%) digestibility. Production performance after the challenge was similar to that before the challenge, and YC improved yield of ECM. After the challenge, supplementing YC tended to reduce rumen lactate concentration compared with the control and reduced haptoglobin in cows fed HS. Feeding HS but not YC increased expression in rumen papillae of genes for receptors (FFAR2 and FFAR3) and transporter (SLC16A3) of short-chain fatty acids but did not affect genes involved in transport of Na⁺/H⁺ or water or in inflammatory response. Supplementing YC to dairy cows improved lactation performance in diets containing low or high starch, and mechanisms might be partially attributed to improvements in rumen pH, digestion of fiber, microbial N synthesis, and reduction in acute phase response.     

Effects of yeast culture supplementation on lactation performance and rumen fermentation profile and microbial abundance in mid-lactation Holstein dairy cows

The continuous trend for a narrowing margin between feed cost and milk prices across dairy farms in the United States highlights the need to improve and maintain feed efficiency. Yeast culture products are alternative supplements that have been evaluated in terms of milk performance and feed efficiency; however, less is known about their potential effects on altering rumen microbial populations and consequently rumen fermentation. Therefore, the objective of this study was to evaluate the effect of yeast culture supplementation on lactation performance, rumen fermentation profile, and abundance of major species of ruminal bacteria in lactating dairy cows. Forty mid-lactation Holstein dairy cows (121 ± 43 days in milk; mean ± standard deviation; 32 multiparous and 8 primiparous) were used in a randomized complete block design with a 7-d adaptation period followed by a 60-d treatment period. Cows were blocked by parity, days in milk, and previous lactation milk yield and assigned to a basal total mixed ration (TMR; 1.6 Mcal/kg of dry matter, 14.6% crude protein, 21.5% starch, and 38.4% neutral detergent fiber) plus 114 g/d of ground corn (CON; n = 20) or basal TMR plus 100 g/d of ground corn and 14 g/d of yeast culture (YC; n = 20; Culture Classic HD, Cellerate Yeast Solutions, Phibro Animal Health Corp.). Treatments were top-dressed over the TMR once a day. Cows were individually fed 1 × /d throughout the trial. Blood and rumen fluid samples were collected in a subset of cows (n = 10/treatment) at 0, 30, and 60 d of the treatment period. Rumen fluid sampled via esophageal tubing was analyzed for ammonia-N, volatile fatty acids (VFA), and ruminal bacteria populations via quantitative PCR amplification of 16S ribosomal DNA genes. Milk yield was not affected by treatment effects. Energy balance was lower in YC cows than CON, which was partially explain by the trend for lower dry matter intake as % body weight in YC cows than CON. Cows fed YC had greater overall ruminal pH and greater total VFA (mM) at 60 d of treatment period. There was a contrasting greater molar proportion of isovalerate and lower acetate proportion in YC-fed cows compared with CON cows. Although the ruminal abundance of specific fiber-digesting bacteria, including Eubacterium ruminantium and Ruminococcus flavefaciens, was increased in YC cows, others such as Fibrobacter succinogenes were decreased. The abundance of amylolytic bacteria such as Ruminobacter amylophilus and Succinimonas amylolytica were decreased in YC cows than CON. Our results indicate that the yeast culture supplementation seems to promote some specific fiber-digesting bacteria while decreasing amylolytic bacteria, which might have partially promoted more neutral rumen pH, greater total VFA, and isovalerate.     

Effects of supplemental yeast (Saccharomyces cerevisiae) culture on rumen development, growth characteristics, and blood parameters in neonatal dairy calves

Yeast (Saccharomyces cerevisiae) culture was added to a texturized calf starter at 0 (control), 1, or 2% of dry matter to determine effects on intake, growth, blood parameters, and rumen development. Seventy-five Holstein calves (38 male; 37 female) were started on the experiment at 2 +/- 1 d of age and were studied for 42 d. Starter intake was measured, and fecal scoring was conducted daily. Growth and blood parameter measurements were recorded at weekly intervals. A subset of 6 male calves (2 per treatment) was euthanized at 5 wk of age, and rumen tissue was sampled for rumen epithelial growth measurements. An additional 6 male calves were euthanized at 6 wk of age for rumen epithelial growth measurements. Inclusion of yeast culture at 2% of the starter ration significantly increased starter and total dry matter intake, average daily gain, and daily hip width change when compared with the control treatment. Average daily gain was improved by 15.6% for the 2% yeast treatment. Daily change in hip height was also significantly greater for calves receiving 2% supplemental yeast culture than for calves receiving 1%. No significant treatment differences were observed for any other variables. These data suggest that the addition of yeast culture in a dairy calf starter at 2% enhances dry matter intake and growth and slightly improves rumen development in dairy calves.     

Analysis of rumen microbial populations in lactating dairy cattle fed diets varying in carbohydrate profiles and Saccharomyces cerevisiae fermentation product

The rumen microbial ecosystem is a critical factor that links diets to bovine physiology and productivity; however, information about dietary effects on microbial populations has generally been limited to small numbers of samples and qualitative assessment. To assess whether consistent shifts in microbial populations occur in response to common dietary manipulations in dairy cattle, samples of rumen contents were collected from 2 studies for analysis by quantitative real-time PCR (qPCR). In one study, lactating Holstein cows (n = 8) were fed diets in which a nonforage fiber source replaced an increasing proportion of forages and concentrates in a 4 × 4 Latin square design, and samples of ruminal digesta were collected at 9-h intervals over 3 d at the end of each period. In the second study, lactating Holstein cows (n = 15) were fed diets with or without the inclusion of a Saccharomyces cerevisiae fermentation product (SCFP) in a crossover design. In this study, rumen liquid and solid samples were collected during total rumen evacuations before and after feeding in a 42-h period. In total, 146 samples of ruminal digesta were used for microbial DNA isolation and analysis by qPCR. Validated primer sets were used to quantify total bacterial and anaerobic fungal populations as well as 12 well-studied bacterial taxa. The relative abundance of the target populations was similar to those previously reported. No significant treatment effects were observed for any target population. A significant interaction of treatment and dry matter intake was observed, however, for the abundance of Eubacterium ruminantium. Increasing dry matter intake was associated with a quadratic decrease in E. ruminantium populations in control animals but with a quadratic increase in E.ruminantium populations in cows fed SCFP. Analysis of sample time effects revealed that Fibrobacter succinogenes and fungal populations were more abundant postfeeding, whereas Ruminococcus albus tended to be more abundant prefeeding. Seven of the target taxa were more abundant in either the liquid or solid fractions of ruminal digesta. By accounting for the total mass of liquid and solid fractions in the rumen and the relative abundance of total bacteria in each fraction, it was estimated that 92% of total bacteria were found in the solid digesta fraction.     

Comparison of fermentation of diets of variable composition and microbial populations in the rumen of sheep and Rusitec fermenters. II. Protozoa population and diversity of bacterial communities

Four ruminally and duodenally cannulated sheep and 8 Rusitec fermenters were used to determine the effects of dietary characteristics on microbial populations and bacterial diversity. The purpose of the study was to assess how closely fermenters can mimic the differences between diets found in vivo. The 4 experimental diets contained forage to concentrate (F:C) ratios of 70:30 (high forage; HF) or 30:70 (high concentrate; HC) with either alfalfa hay (A) or grass hay (G) as the forage. Total bacterial numbers were greater in the rumen of sheep fed HF diets compared with those fed HC diets, whereas the opposite was found in fermenters. The numbers of cellulolytic bacteria were not affected by F:C ratio in any fermentation system, but cellulolytic numbers were 2.7 and 1.8 times greater in sheep than in fermenters for HF and HC diets, respectively. Neither total bacterial nor cellulolytic numbers were affected by the type of forage in sheep or fermenters. Decreasing F:C ratio increased total protozoa and Entodiniae numbers in sheep by about 29 and 25%, respectively, but it had no effect in fermenters. Isotrichidae and Ophryoscolecinae numbers in sheep were not affected by changing F:C ratio, but both disappeared completely from fermenters fed HC diets. Total protozoa and Entodiniae numbers were greater in sheep fed A diets than in those fed G diets, whereas the opposite was found in fermenters. Results indicate that under the conditions of the present study, protozoa population in Rusitec fermenters was not representative of that in the rumen of sheep fed the same diets. In addition, protozoa numbers in fermenters were 121 and 226 times lower than those in the sheep rumen for HF and HC diets, respectively. The automated ribosomal intergenic spacer analysis of the 16S ribosomal DNA was used to analyze the diversity of liquid- and solid-associated bacteria in both systems. A total of 170 peaks were detected in the automated ribosomal intergenic spacer analysis electropherograms of bacterial pellets across the full set of 64 samples, from which 160 were detected in at least 1 individual from each system (sheep or fermenter). Diversity of liquid-associated bacterial pellets was greater with G diets in fermenters but seemed to be unaffected by diet in sheep. Bacterial diversity in solid-associated bacteria pellets was greater for G diets compared with A diets in sheep and fermenters. Different conditions in the fermenters compared with sheep rumen might have caused a selection of some bacterial strains.