Effect of dietary concentrate on rumen fermentation, digestibility, and nitrogen losses in dairy cows

The objective of this experiment was to investigate the effect of level of dietary concentrate on rumen fermentation, digestibility, and N losses in lactating dairy cows. The experiment was a replicated 3 × 3 Latin square design with 6 cows and 16-d adaptation periods. Ruminal contents were exchanged between cows at the beginning of each adaptation period. Data for 2 of the diets tested in this experiment are presented here. The diets contained (dry matter basis): 52% (LowC; control) and 72% (HighC) concentrate feeds. Crude protein contents of the diets were 16.5 and 16.4%, respectively. The HighC diet decreased ruminal pH and ammonia concentration and increased propionate concentration compared with LowC. Acetate:propionate ratio was greater for LowC than for HighC. Rumen methane production and microbial protein synthesis were unaffected by diet. Dry matter intake was similar among diets, but milk yield was increased by HighC compared with LowC (36.0 and 33.2 kg/d, respectively). Milk fat percentage and yield and total-tract apparent NDF digestibility were decreased by HighC compared with LowC. More ruminal ammonia N was transferred into milk protein with HighC than with LowC. Urinary N excretion, plasma urea N, and milk urea N concentration were not affected by diet. The ammonia emitting potential of manure was similar between LowC and HighC diets. Increased concentrate proportion in the diet of dairy cows resulted in reduced ruminal ammonia concentration and enhanced ammonia utilization for milk protein synthesis. These effects, however, did not result in reduced urinary N losses and only marginally improved milk N efficiency. Increasing dietary concentrate was not a successful strategy to mitigate enteric methane production and ammonia emissions from manure.     

Nitrate improves ammonia incorporation into rumen microbial protein in lactating dairy cows fed a low-protein diet

Generation of ammonia from nitrate reduction is slower compared with urea hydrolysis and may be more efficiently incorporated into ruminal microbial protein. We hypothesized that nitrate supplementation could increase ammonia incorporation into microbial protein in the rumen compared with urea supplementation of a low-protein diet fed to lactating dairy cows. Eight multiparous Chinese Holstein dairy cows were used in a crossover design to investigate the effect of nitrate or an isonitrogenous urea inclusion in the basal low-protein diet on rumen fermentation, milk yield, and ruminal microbial community in dairy cows fed a low-protein diet in comparison with an isonitrogenous urea control. Eight lactating cows were blocked in 4 pairs according to days in milk, parity, and milk yield and allocated to urea (7.0 g urea/kg of dry matter of basal diet) or nitrate (14.6 g of NO₃^?/kg of dry matter of basal diet, supplemented as sodium nitrate) treatments, which were formulated on 75% of metabolizable protein requirements. Nitrate supplementation decreased ammonia concentration in the rumen liquids (?33.1%) and plasma (?30.6%) as well as methane emissions (?15.0%) and increased dissolved hydrogen concentration (102%), microbial N (22.8%), propionate molar percentage, milk yield, and 16S rRNA gene copies of Selenomonas ruminantium. Ruminal dissolved hydrogen was positively correlated with the molar proportion of propionate (r = 0.57), and negatively correlated with acetate-to-propionate ratio (r = ?0.57) and estimated net metabolic hydrogen production relative to total VFA produced (r = ?0.58). Nitrate reduction to ammonia redirected metabolic hydrogen away from methanogenesis, enhanced ammonia incorporation into rumen microbial protein, and shifted fermentation from acetate to propionate, along with increasing S. ruminantium 16S rRNA gene copies, likely leading to the increased milk yield.     

Effect of Saccharomyces cerevisiae fermentation product on ruminal fermentation and nutrient utilization in dairy cows

The goal of this experiment was to investigate the effect of yeast culture (Saccharomyces cerevisiae) on rumen fermentation, nutrient utilization, and ammonia and methane emission from manure in dairy cows. Eight ruminally cannulated Holstein cows were allocated to 2 dietary treatments in a crossover design. Treatments were control (no yeast culture) and XP (yeast culture, fed at 56 g/head per day; XP, Diamond V Mills Inc., Cedar Rapids, IA). Dry matter intake, milk yield, milk composition, and body weight were similar between treatments. Milk urea nitrogen concentration was also not affected by treatment. Rumen pH was similar between the control and XP treatments, but rumen ammonia concentration tended to be lower with XP than with the control. Treatment had no effect on concentrations of total or individual volatile fatty acids, protozoal counts, polysaccharide-degrading activities (except amylase activity that tended to be increased by XP), or methane production in the rumen. Urinary N losses did not differ significantly between treatments, but allantoin and total purine derivative excretions and the estimated microbial N outflow from the rumen tended to be increased by XP compared with the control treatment. Total-tract apparent digestibility of dietary nutrients was not affected by XP. Milk fatty acid composition was also not altered by XP supplementation. Cumulative (253 h) ammonia and methane emissions from manure, measured in a steady-state gas emission system, were slightly decreased by XP. Overall, the yeast culture tested had little effect on ruminal fermentation, digestibility, or N losses, but tended to reduce rumen ammonia concentration and increase microbial protein synthesis in the rumen, and decreased ammonia and methane emissions from manure.     

Ginkgo fruit extract as an additive to modify rumen microbiota and fermentation and to mitigate methane production

Ginkgo fruit, an unused byproduct of the ginkgo nut industry, contains antimicrobial compounds known as anacardic acids. Two major cultivars of ginkgo, Kyuju (K) and Tokuro (T), were evaluated for their potential as a feed additive for ruminants. In batch culture, we incubated a mixture of hay and concentrate in diluted rumen fluid with or without 1.6% (fruit equivalent) ginkgo fruit extract. We conducted another series of batch culture studies to determine the dose response of fermentation. We also conducted continuous culture using the rumen simulation technique (RUSITEC) with cultivar K and carried out a pure culture study to monitor the sensitivity of 17 representative rumen bacterial species to ginkgo extract and component phenolics. Although both K and T extracts led to decreased methane and increased propionate production, changes were more apparent with K extract, and were dose-dependent. Total gas production was depressed at doses ≥3.2%, suggesting that 1.6% was the optimal supplementation level. In RUSITEC fermentation supplemented with 1.6% ginkgo K, methane decreased by 53% without affecting total gas or total VFA production, but with decreased acetate and increased propionate. Disappearance of dry matter, neutral detergent fiber, and acid detergent fiber were not affected by ginkgo, but ammonia levels were decreased. Quantitative PCR indicated that the abundance of protozoa, fungi, methanogens, and bacteria related to hydrogen and formate production decreased, but the abundance of bacteria related to propionate production increased. MiSeq analysis (Illumina Inc., San Diego, CA) confirmed these bacterial changes and identified archaeal community changes, including a decrease in Methanobrevibacter and Methanomassiliicoccaceae and an increase in Methanoplanus. Pure culture study results supported the findings for the above bacterial community changes. These results demonstrate that ginkgo fruit can modulate rumen fermentation toward methane mitigation and propionate enhancement via microbial selection.     

Effects of 3-nitrooxypropanol on methane emission,digestion, and energy and nitrogen balance of lactating dairy cows

The objective was to measure effects of 3-nitrooxypropanol (3NP) on methane production of lactating dairy cows and any associated changes in digestion and energy and N metabolism. Six Holstein-Friesian dairy cows in mid-lactation were fed twice daily a total mixed ration with maize silage as the primary forage source. Cows received 1 of 3 treatments using an experimental design based on two 3 × 3 Latin squares with 5-wk periods. Treatments were a control placebo or 500 or 2,500 mg/d of 3NP delivered directly into the rumen, via the rumen fistula, in equal doses before each feeding. Measurements of methane production and energy and N balance were obtained during wk 5 of each period using respiration calorimeters and digestion trials. Measurements of rumen pH (48 h) and postprandial volatile fatty acid and ammonia concentrations were made at the end of wk 4. Daily methane production was reduced by 3NP, but the effects were not dose dependent (reductions of 6.6 and 9.8% for 500 and 2,500 mg/d, respectively). Dosing 3NP had a transitory inhibitory effect on methane production, which may have been due to the product leaving the rumen in liquid outflow or through absorption or metabolism. Changes in rumen concentrations of volatile fatty acids indicated that the pattern of rumen fermentation was affected by both doses of the product, with a decrease in acetate:propionate ratio observed, but that acetate production was inhibited by the higher dose. Dry matter, organic matter, acid detergent fiber, N, and energy digestibility were reduced at the higher dose of the product. The decrease in digestible energy supply was not completely countered by the decrease in methane excretion such that metabolizable energy supply, metabolizable energy concentration of the diet, and net energy balance (milk plus tissue energy) were reduced by the highest dose of 3NP. Similarly, the decrease in N digestibility at the higher dose of the product was associated with a decrease in body N balance that was not observed for the lower dose. Milk yield and milk fat concentration and fatty acid composition were not affected but milk protein concentration was greater for the higher dose of 3NP. Twice-daily rumen dosing of 3NP reduced methane production by lactating dairy cows, but the dose of 2,500 mg/d reduced rumen acetate concentration, diet digestibility, and energy supply. Further research is warranted to determine the optimal dose and delivery method of the product.     

Diet supplementation with thyme oil and its main component thymol failed to favorably alter rumen fermentation,improve nutrient utilization,or enhance milk production in dairy cows

Phenolic compounds and essential oils with high content of phenolic compounds have been reported to exert antimicrobial activities in vitro. The objective of this study was to determine the effects of dairy cow diet supplementation with thyme oil and its main component thymol on intake and total-tract apparent digestibility of nutrients, rumen fermentation characteristics, ruminal protozoa, nitrogen excretion, and milk production. For this aim, we used 8 multiparous, ruminally cannulated Holstein cows in a replicated 4 × 4 Latin square design (28 d periods), balanced for residual effects. Cows were fed 1 of the 4 following experimental treatments: total mixed ration (TMR) with no additive (control); TMR + monensin [24 mg/kg of dry matter (DM)]; TMR + thyme oil (50 mg/kg of DM); and TMR + thymol (50 mg/kg of DM). Compared with the control diet, feeding thyme oil or thymol had no effect on DM intake, nutrient total-tract apparent digestibility, total N excretion, ruminal pH, ammonia concentration, total volatile fatty acid (VFA) concentration, or acetate:propionate ratio. Ruminal protozoa density was not modified by thyme oil, but decreased with thymol supplementation. Supplementation with thyme oil or thymol did not affect milk production, milk composition, or efficiency of milk production. Neither thyme oil nor thymol affected efficiency of dietary N use for milk N secretion (N intake/milk N). Supplementation with monensin tended to decrease DM intake (–1.2 kg/d) and milk fat yield. Total-tract apparent digestibility of nutrients did not differ between cows fed monensin and cows fed the control diet. Total VFA concentration was not changed by monensin supplementation compared with control, but adding monensin shifted the VFA profile toward more propionate and less acetate, resulting in a decrease of acetate:propionate ratio. Protozoa density and ammonia concentration were lower in the ruminal content of cows fed monensin compared with that of cows fed the control diet. Total N excretion was not affected by monensin supplementation. Likewise, efficiency of use of dietary N for milk N secretion was unchanged in cows fed monensin. The results of this study contrasted with the claimed in vitro antimicrobial activity of thyme oil and thymol: we observed no positive effects on rumen metabolism (i.e., N and VFA) or milk performance in dairy cows. Under the conditions of this study, including thyme oil or thymol at 50 mg/kg of DM had no benefits for rumen fermentation, nutrient utilization and milk performance in dairy cows.     

Effect of dietary carbohydrate composition and availability on utilization of ruminal ammonia nitrogen for milk protein synthesis in dairy cows

A trial with four ruminally and duodenally cannulated, late-lactation dairy cows was conducted to investigate the effect of dietary carbohydrate (CHO) composition and availability on ruminal ammonia N utilization and transfer into milk protein. Two diets were fed at 8-h intervals in a crossover design. The diets differed in CHO composition: the ruminally fermentable non-structural carbohydrates (RFSS) diet (barley and molasses) contained a larger proportion of ruminally available CHO in the nonstructural carbohydrate fractions and the ruminally fermentable fiber (RFNDF) diet (corn, beet pulp, and brewer's grains) contained a larger proportion of CHO in ruminally available fiber. Nitrogen-15 was used to label ruminal ammonia N and consequently microbial and milk N. Fermentation acids, enzyme activities, and microbial protein production in the rumen were not affected by diet. Ruminal ammonia concentration was lowered by RFNDF. Ruminal and total tract digestibility of nutrients did not differ between diets except that apparent ruminal degradability of crude protein was lower for RFNDF compared with RFSS. Partitioning of N losses between urine and feces was also not affected by diet. Milk yield and fat and protein content were not affected by treatment. Average concentration of milk urea N was lower for RFNDF than for RFSS. Proportion of milk protein N originating from ruminal microbial N (based on the areas under the 15N-enrichment curves) was higher for RFNDF than for RFSS. Cumulative recovery of 15N in milk protein was 13% higher for RFNDF than for RFSS indicating enhanced transfer of 15N-ammonia into milk protein with the former diet. The results suggested that, compared to diets containing higher levels of ruminally fermentable starch, diets providing higher concentration of ruminally fermentable fiber may enhance transfer of ruminal ammonia and microbial N into milk protein.     

Effect of essential oil active compounds on rumen microbial fermentation and nutrient flow in in vitro systems

Two experiments were conducted to determine the effects of several essential oil active compounds on rumen microbial fermentation. In the first experiment, 4 doses (5, 50, 500, and 5,000 mg/L) of 5 essential oil compounds were evaluated using in vitro 24-h batch culture of rumen fluid with a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were control (CON), eugenol (EUG), guaiacol, limonene, thymol (THY), and vanillin. After 24 h, the pH was determined, and samples were collected to analyze ammonia N and volatile fatty acids (VFA). The highest dose of all compounds decreased total VFA concentration and increased the final pH. Eugenol at 5 mg/L tended to reduce the proportion of acetate and the acetate to propionate ratio, at 50 and 500 mg/L tended to reduce ammonia N concentration, and at 500 mg/L reduced the proportion of propionate and branched-chain VFA concentration, without affecting total VFA concentration. All other treatments had minor effects or changes occurred only after total VFA concentration decreased. In the second experiment, 8 dual-flow continuous culture fermenters (1,320 mL) were used in 3 replicated periods (6 d of adaptation and 3 d of sampling) to study the effects of THY and EUG on rumen microbial fermentation. Fermenters were fed 95 g/d of DM of a 60:40 forage:concentrate diet (18% crude protein; 30% neutral detergent fiber). Treatments were CON, 10 mg/L of monensin (positive control), and 5, 50, or 500 mg/L of THY and EUG, and were randomly assigned to fermenters within periods. During the last 3 d of each period, samples were taken at 0, 2, 4, and 6 h after the morning feeding and analyzed for peptides, amino acids, and ammonia N concentrations, and total and individual VFA concentrations. Monensin changed the VFA profile as expected, but inhibited nutrient digestion. Eugenol and THY decreased total VFA concentration and changed the VFA profile, and only 5 mg/L of THY tended to reduce the proportion of acetate, increased the proportion of butyrate, and increased the large peptides N concentration without decreasing total VFA concentration. Most of these essential oil compounds demonstrated their antimicrobial activity by decreasing total VFA concentration at high doses. However, EUG in batch fermentation and 5 mg/L of THY in continuous culture modified the VFA profile without decreasing total VFA concentration, and EUG in batch fermentation decreased ammonia N concentration.     

Effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and nutrient utilization in dairy does

The objective of this study was to evaluate effects of fumarate on ruminal ammonia accumulation and fiber digestion in vitro and on feed intake and nutrient utilization in dairy does. Batch cultures of mixed rumen microorganisms were used to study effects of different concentrations of fumarate on fermentation with various N sources (ammonia as ammonium bicarbonate, casein amino acids, casein peptides, gelatin peptides) and feeds (bermudagrass hay, mixed diet of 60% bermudagrass hay plus 40% concentrate) for 6 and 24 h, respectively. Substrates were grouped into pairs for separate incubations. Monosodium fumarate was added to incubation tubes to achieve final concentrations of 0, 5, and 10 mM fumarate. More ammonia accumulated at the end of incubation with added ammonium bicarbonate. Ammonia concentration was higher for peptide compared with amino acid incubation, and for casein peptide compared with gelatin peptide. Addition of fumarate linearly decreased ammonia for all N sources and for feed substrates. For all substrate types, fumarate treatment increased acetate, propionate, and total volatile fatty acids (VFA), decreased acetate to propionate ratio, and tended to reduce branched-chain VFA. Digestion of feed neutral detergent fiber (NDF) by rumen microorganisms was improved by fumarate along with elevated endoglucanase and xylanase activities. In an animal metabolism experiment, 8 dairy does (4 per treatment) were used in a completely randomized design for 21 d. Does were fed a hay plus concentrate diet without (control) or with fumarate (6 g/head per day) supplementation to determine feed intake, whole-tract nutrient digestibility, and N utilization. Fumarate treatment did not affect weight change or feed intake but increased whole-tract digestion of gross energy, crude protein, and cellulose. Digested N was increased by fumarate supplementation; however, N retention was unaffected. Plasma glucose concentration was elevated with fumarate but urea N concentration remained unchanged. Fumarate addition had significant effects on rumen microbial fermentation by decreasing ammonia and branched-chain VFA, and by increasing acetate and propionate, and NDF digestion. These effects were reflected in the improvement in whole-tract gross energy, crude protein, and cellulose digestion and elevated plasma glucose concentration when dairy does were supplemented with fumarate.     

Effect of a garlic and citrus extract supplement on performance,rumen fermentation,methane production,and rumen microbiome of dairy cows

The aim of this trial was to determine the effect of a garlic and citrus extract supplement (GCE) on the performance, rumen fermentation, methane emissions, and rumen microbiome of dairy cows. Fourteen multiparous Nordic Red cows in mid-lactation from the research herd of Luke (Jokioinen, Finland) were allocated to 7 blocks in a complete randomized block design based on body weight, days in milk, dry matter intake (DMI), and milk yield. Animals within each block were randomly allocated to a diet with or without GCE. The experimental period for each block of cows (one for each of the control and GCE groups) consisted of 14 d of adaptation followed by 4 d of methane measurements inside the open circuit respiration chambers, with the first day being considered as acclimatization. Data were analyzed using the GLM procedure of SAS (SAS Institute Inc.). Methane production (g/d) and methane intensity (g/kg of energy-corrected milk) were lower by 10.3 and 11.7%, respectively, and methane yield (g/kg of DMI) tended to be lower by 9.7% in cows fed GCE compared with the control. Dry matter intake, milk production, and milk composition were similar between treatments. Rumen pH and total volatile fatty acid concentrations in rumen fluid were similar, whereas GCE tended to increase molar propionate concentration and decrease the molar ratio of acetate to propionate. Supplementation with GCE resulted in greater abundance of Succinivibrionaceae, which was associated with reduced methane. The relative abundance of the strict anaerobic Methanobrevibacter genus was reduced by GCE. The change in microbial community and rumen propionate proportion may explain the decrease in enteric methane emissions. In conclusion, feeding GCE to dairy cows for 18 d modified rumen fermentation and microbiota, leading to reduced methane production and intensity without compromising DMI or milk production in dairy cows. This could be an effective strategy for enteric methane mitigation of dairy cows.     

Nutrient digestion and lactation performance of dairy cows fed combinations of prilled fat and canola oil.

Four combinations of prilled fat and canola oil were fed to 10 lactating Holstein cows in a replicated 5 x 5 Latin square to determine whether mixing plant oil with a rumen inert fat had additive effects on digestive and lactation responses. Five diets of concentrate and corn silage (1:1, DM basis) contained either no added fat (control) or 5% fat comprising 100, 67, 33, or 0% prilled fat and the remainder canola oil. The fat supplement containing 100% prilled fat appeared to be rumen-inert because it caused no changes in ruminal VFA concentration, acetate to propionate ratio, or total tract fiber digestion. Prilled fat increased milk production, FCM, and milk fat percentage but decreased milk protein percentage, including casein content. Increasing canola oil in the fat supplement caused linear declines in ruminal VFA, acetate to propionate ratio, and milk production. Milk production efficiency (weight FCM/weight DMI) exceeded the control diet when fat supplements contained 100 or 67% prilled fat but dropped below control for 33 and 0% prilled fat. This study demonstrates additive effects of combining canola oil with hydrogenated, prilled fat on ruminal fermentation but nonadditive effects on milk production efficiency and milk composition. At low levels of supplementation, plant oils, such as the canola oil used in this study, can inhibit ruminal fermentation but still maintain milk production efficiency.     

Plant extracts affect in vitro rumen microbial fermentation

Different doses of 12 plant extracts and 6 secondary plant metabolites were incubated for 24 h in diluted ruminal fluid with a 50:50 forage:concentrate diet. Treatments were: control (no additive), plant extracts (anise oil, cade oil, capsicum oil, cinnamon oil, clove bud oil, dill oil, fenugreek, garlic oil, ginger oil, oregano oil, tea tree oil, and yucca), and secondary plant metabolites (anethol, benzyl salicylate, carvacrol, carvone, cinnamaldehyde, and eugenol). Each treatment was supplied at 3, 30, 300, and 3,000 mg/L of culture fluid. At 3,000 mg/L, most treatments decreased total volatile fatty acid concentration, but cade oil, capsicum oil, dill oil, fenugreek, ginger oil, and yucca had no effect. Different doses of anethol, anise oil, carvone, and tea tree oil decreased the proportion of acetate and propionate, which suggests that these compounds may not be nutritionally beneficial to dairy cattle. Garlic oil (300 and 3,000 mg/L) and benzyl salicylate (300 and 3,000 mg/L) reduced acetate and increased propionate and butyrate proportions, suggesting that methane production was inhibited. At 3,000 mg/L, capsicum oil, carvacrol, carvone, cinnamaldehyde, cinnamon oil, clove bud oil, eugenol, fenugreek, and oregano oil resulted in a 30 to 50% reduction in ammonia N concentration. Careful selection and combination of these extracts may allow the manipulation of rumen microbial fermentation.     

Between-cow variation in digestion and rumen fermentation variables associated with methane production

A meta-analysis based on an individual-cow data set was conducted to investigate the effects of between-cow variation and related animal variables on predicted CH₄ emissions from dairy cows. Data were taken from 40 change-over studies consisting of a total of 637 cow/period observations. Animal production and rumen fermentation characteristics were measured for 154 diets in 40 studies; diet digestibility was measured for 135 diets in 34 studies, and ruminal digestion kinetics was measured for 56 diets in 15 studies. The experimental diets were based on grass silage, with cereal grains or by-products as energy supplements, and soybean or canola meal as protein supplements. Average forage:concentrate ratio across all diets on a dry matter basis was 59:41. Methane production was predicted from apparently fermented substrate using stoichiometric principles. Data were analyzed by mixed-model regression using diet and period within experiment as random effects, thereby allowing the effect of experiment, diet, and period to be excluded. Dry matter intake and milk yield were more repeatable experimental measures than rumen fermentation, nutrient outflow, diet digestibility, or estimated CH₄ yield. Between-cow coefficient of variation (CV) was 0.010 for stoichiometric CH₄ per mol of volatile fatty acids and 0.067 for predicted CH₄ yield (CH₄/dry matter intake). Organic matter digestibility (OMD) also displayed little between-cow variation (CV = 0.013), indicating that between-cow variation in diet digestibility and rumen fermentation pattern do not markedly contribute to between cow-variation in CH₄ yield. Digesta passage rate was much more variable (CV = 0.08) between cows than OMD or rumen fermentation pattern. Increased digesta passage rate is associated with improved energetic efficiency of microbial N synthesis, which partitions fermented substrate from volatile fatty acids and gases to microbial cells that are more reduced than fermented carbohydrates. Positive relationships were observed between CH₄ per mol of volatile fatty acids versus OMD and rumen ammonia N concentration versus OMD; and negative relationships between the efficiency of microbial N synthesis versus OMD and digesta passage rate versus OMD, suggesting that the effects of these variables on CH₄ yield were additive. It can be concluded that variations in OMD and efficiency in microbial N synthesis resulting from variations in digesta passage contribute more to between-animal variation in CH₄ emissions than rumen fermentation pattern.     

Balancing carbohydrates and proteins for optimum rumen microbial yield.

Establishing conditions under which rumen fermentation will be optimized requires an understanding of the nutrient requirements of the mixed microbial population. The major nutrients required by rumen microbes are carbohydrates and proteins, but the most suitable sources and quantities needed to support maximum growth have not been determined. Digestion of proteins results in the production of peptides, which can accumulate in the rumen. Peptides are further hydrolyzed to amino acids, some of which are deaminated, producing ammonia. Although peptides, amino acids, and ammonia all may individually serve as sources of N for various microbes, the total population achieves the highest growth rate on mixtures of all three sources. In a somewhat analogous manner, carbohydrates are digested by exoenzymes to oligosaccharides that are available for crossfeeding by the mixed microbial population. Based on data from both in vitro and in vivo studies, there is general agreement that rate of digestion of carbohydrates is the major factor controlling the energy available for microbial growth; in addition, rate of digestion of total carbohydrate is directly related to proportion of starches, pectins, and sugars. Proteins affect both total fermentation and production of microbial DM per unit of carbohydrate fermented. It appears that the quantity of ruminally available protein needed to optimize microbial growth may, under some conditions, be as high as 14 to 15% of diet DM.     

Effect of garlic oil and four of its compounds on rumen microbial fermentation

Different concentrations (3, 30, 300, and 3000 mg/L of culture fluid) of garlic oil (GAR), diallyl sulfide (DAS), diallyl disulfide (DAD), allicin (ALL), and allyl mercaptan (ALM) were incubated for 24 h in diluted ruminal fluid with a 50:50 forage:concentrate diet (17.7% crude protein; 30.7% neutral detergent fiber) to evaluate their effects on rumen microbial fermentation. Garlic oil (30 and 300 mg/L), DAD (30 and 300 mg/L), and ALM (300 mg/L) resulted in lower molar proportion of acetate and higher proportions of propionate and butyrate. In contrast, at 300 mg/L, DAS only increased the proportion of butyrate, and ALL had no effects on volatile fatty acid proportions. In a dual-flow continuous culture of rumen fluid fed the same 50:50 forage:concentrate diet, addition of GAR (312 mg/L), DAD (31.2 and 312 mg/L), and ALM (31.2 and 312 mg/L) resulted in similar changes to those observed in batch culture, with the exception of the lack of effect of DAD on the proportion of propionate. In a third in vitro study, the potential of GAR (300 mg/L), DAD (300 mg/L), and ALM (300 mg/L) to decrease methane production was evaluated. Treatments GAR, DAD, and ALM resulted in a decrease in methane production of 73.6, 68.5, and 19.5%, respectively, compared with the control. These results confirm the ability of GAR, DAD, and ALM to decrease methane production, which may help to improve the efficiency of energy use in the rumen.     

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.     

Microbial protein synthesis and flows of nitrogen fractions to the duodenum of dairy cows.

Attempts have been made to increase nutrient availability for milk production by increasing feed intake, optimizing ruminal fermentation, and supplementing nutrients to the diet that will escape ruminal degradation. Energy and N are the nutritional factors that most often limit microbial growth and milk production. Ruminal fermentation and flow of microbial and dietary protein to the small intestine are affected by feed intake and by the amount and source of energy and protein in the diet. Feeding protein and carbohydrate that are not degraded in the rumen increases the quantity of dietary protein that passes to the small intestine but may decrease the quantity of microbial protein that is synthesized in the rumen. This often results in only small differences in the total NAN that passes to the small intestine. Because microbial protein supplies a large quantity of total AA that passes to the small intestine, differences in passage of individual AA often are only slight. Additional research with cows consuming large amounts of feed are needed to identify combinations of feed ingredients that synchronize availabilities of energy and N for optimizing ruminal digestion, microbial protein synthesis, nutrient flow to the small intestine, and milk production and composition.     

Effects of antibiotics and oil on microbial profiles and fermentation in mixed cultures of ruminal microorganisms

Ionophores and supplemental fat are fed to lactating cows to improve feed efficiency. Their effect on rumen fermentation is similar, but less is known about their impact on rumen microbes. The objective of this study was to determine the effects of monensin (M), bacitracin (B), and soybean oil (O) on microbial populations. Mixed cultures of rumen microbes were incubated in 5 dual-flow continuous fermentors and fed 13.8 g of alfalfa hay pellets daily (DM basis) for 16 d. All fermentors were allowed to stabilize for 4 d. From d 5 to 10, two fermentors received O (5% of diet DM), one fermentor received M (22 mg/kg), and one received B (22 mg/kg). From d 11 to 16, the 2 fermentors receiving O also received either M (OM) or B (OB) and O was included in the fermentors receiving M (MO) and B (BO). One fermentor served as the control and received 100% alfalfa pellets throughout the experiment. Each run was replicated 3 times. Samples were taken at 2 h after the morning feeding on d 4, 10, and 16 and were analyzed for bacterial populations using terminal restriction fragment length polymorphism. Volatile fatty acid concentration, methane production, and pH in the control cultures were not affected by time and remained similar during the entire experiment. The M and O treatments reduced molar concentration of acetate, increased concentration of propionate, and decreased methane production. Bacitracin did not alter acetate or propionate concentration, but reduced methane production. All 3 treatments (M, B, and O) altered the fragment patterns of microbial profiles. In contrast, treatments MO, OM, BO, and OB had little effect on culture fermentation despite differences in the patterns of microbial fragments. The terminal restriction fragment length polymorphism data suggest that microbial adaptation to the in vitro system in the control fermentor occurred within 4 d.     

Plant oil supplements reduce methane emissions and improve milk fatty acid composition in dairy cows fed grass silage-based diets without affecting milk yield

Four lipid supplements varying in chain length or degree of unsaturation were examined for their effects on milk yield and composition, ruminal CH₄ emissions, rumen fermentation, nutrient utilization, and microbial ecology in lactating dairy cows. Five Nordic Red cows fitted with rumen cannulas were used in a 5 × 5 Latin square with five 28-d periods. Treatments comprised total mixed rations based on grass silage with a forage-to-concentrate ratio of 60:40 supplemented with no lipid (CO) or 50 g/kg of diet dry matter (DM) of myristic acid (MA), rapeseed oil (RO), safflower oil (SO), or linseed oil (LO). Feeding MA resulted in the lowest DM intake, and feeding RO reduced DM intake compared with CO. Feeding MA reduced the yields of milk, milk constituents, and energy-corrected milk. Plant oils did not influence yields of milk and milk constituents, but reduced milk protein content compared with CO. Treatments had no effect on rumen fermentation characteristics, other than an increase in ammonia-N concentration due to feeding MA, RO, and SO compared with CO. Lipid supplements reduced daily ruminal CH₄ emission; however, the response was to some extent a result of lower feed intake. Lipids modified microbial community structure without affecting total counts of bacteria, archaea, and ciliate protozoa. Dietary treatments had no effect on the apparent total tract digestibility of organic matter, fiber, and gross energy. Treatments did not affect either energy secreted in milk as a proportion of energy intake or efficiency of dietary N utilization. All lipids lowered de novo fatty acid synthesis in the mammary gland. Plant oils increased proportions of milk fat 18:0, cis 18:1, trans and monounsaturated fatty acids, and decreased saturated fatty acids compared with CO and MA. Both SO and LO increased the proportion of total polyunsaturated fatty acids, total conjugated linolenic acid, and cis-9,trans-11 conjugated linoleic acid. Feeding MA clearly increased the Δ⁹ desaturation of fatty acids. Our results provide compelling evidence that plant oils supplemented to a grass silage-based diet reduce ruminal CH₄ emission and milk saturated fatty acids, and increase the proportion of unsaturated fatty acids and total conjugated linoleic acid while not interfering with digestibility, rumen fermentation, rumen microbial quantities, or milk production.     

Effects of calcium salts of fatty acids and proportion of forage in diet on ruminal fermentation and nutrient flow to duodenum of cows.

Four Holstein cows fitted with ruminal and duodenal cannulas were used in a 4 x 4 Latin square to investigate the effects of calcium salts of long-chain fatty acids (fat) and proportion of forage in diet on ruminal fermentation, flow of nutrients to the small intestine, and animal performance. Treatments, arranged in a 2 x 2 (fat x forage) factorial, were 1) low (50%) forage, no fat; 2) low forage, fat; 3) high (67%) forage, no fat; and 4) high forage, fat. Feeding fat decreased OM intake and OM truly digested in the rumen. Feeding high forage diets decreased intakes of OM and starch and increased intakes of ADF and NDF. Ruminal pH and ratio of acetate to propionate were increased with high forage diets compared with low forage diets. Feeding fat and different amounts of forage to cows did not alter the flows of NAN and microbial N to the duodenum or efficiency of microbial growth. Production of milk and 4% FCM and percentage of fat in milk were increased by feeding fat. Feeding high forage diets decreased milk production, increased percentage of fat in milk, increased the yield of fat, and caused no change in 4% FCM production. The percentage of protein in milk was decreased by feeding high forage diets and fat, but yield of milk protein was decreased only by feeding high forage diets to cows.