The goal of this review was to analyze published data related to mitigation of enteric methane (CH4) emissions from ruminant animals to document the most effective and sustainable strategies. Increasing forage digestibility and digestible forage intake was one of the major recommended CH4 mitigation practices. Although responses vary, CH4 emissions can be reduced when corn silage replaces grass silage in the diet. Feeding legume silages could also lower CH4 emissions compared to grass silage due to their lower fiber concentration. Dietary lipids can be effective in reducing CH4 emissions, but their applicability will depend on effects on feed intake, fiber digestibility, production, and milk composition. Inclusion of concentrate feeds in the diet of ruminants will likely decrease CH4 emission intensity (Ei; CH4 per unit animal product), particularly when inclusion is above 40% of dietary dry matter and rumen function is not impaired. Supplementation of diets containing medium to poor quality forages with small amounts of concentrate feed will typically decrease CH4 Ei. Nitrates show promise as CH4 mitigation agents, but more studies are needed to fully understand their impact on whole-farm greenhouse gas emissions, animal productivity, and animal health. Through their effect on feed efficiency and rumen stoichiometry, ionophores are likely to have a moderate CH4 mitigating effect in ruminants fed high-grain or mixed grain-forage diets. Tannins may also reduce CH4 emissions although in some situations intake and milk production may be compromised. Some direct-fed microbials, such as yeast-based products, might have a moderate CH4-mitigating effect through increasing animal productivity and feed efficiency, but the effect is likely to be inconsistent. Vaccines against rumen archaea may offer mitigation opportunities in the future although the extent of CH4 reduction is likely to be small and adaptation by ruminal microbes and persistence of the effect is unknown. Overall, improving forage quality and the overall efficiency of dietary nutrient use is an effective way of decreasing CH4 Ei. Several feed supplements have a potential to reduce CH4 emission from ruminants although their long-term effect has not been well established and some are toxic or may not be economically feasible.

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Currently used microbial markers cannot distinguish protozoal nitrogen (N) from bacterial N, thus limiting research on protozoal quantification in vivo by the lack of a repeatable, accurate marker for protozoal N. We report the development of a real-time PCR assay targeting the gene encoding 18S rDNA to quantify the amount of protozoal biomass in ruminal fluid and duodenal digesta. Protozoal cells were harvested from rumen fluid and concentrated for evaluation of recovery of rDNA in samples from the rumen and the duodenum. The DNA from concentrated cells was extracted with virtually 100% efficiency both before and after column purification. After serial spiking of protozoal cells into duodenal fluid over the entire range of quantification, the recovery was highly linear and constant at 81%. After serially spiking increasing quantities of protozoal rDNA into a constant volume of duodenal samples, nonlinear regression verified constant recovery of background rDNA in duodenal samples regardless of the ratio of target:nontarget rDNA. Recommendations for the procedure, including replication per sample, are described herein.

Although livestock production accounts for a sizeable share of global greenhouse gas emissions, numerous technical options have been identified to mitigate these emissions. In this review, a subset of these options, which have proven to be effective, are discussed. These include measures to reduce CH 4 emissions from enteric fermentation by ruminants, the largest single emission source from the global livestock sector, and for reducing CH 4 and N 2 O emissions from manure. A unique feature of this review is the high level of attention given to interactions between mitigation options and productivity. Among the feed supplement options for lowering enteric emissions, dietary lipids, nitrates and ionophores are identified as the most effective. Forage quality, feed processing and precision feeding have the best prospects among the various available feed and feed management measures. With regard to manure, dietary measures that reduce the amount of N excreted (e.g. better matching of dietary protein to animal needs), shift N excretion from urine to faeces (e.g. tannin inclusion at low levels) and reduce the amount of fermentable organic matter excreted are recommended. Among the many 'end-of-pipe' measures available for manure management, approaches that capture and/or process CH 4 emissions during storage (e.g. anaerobic digestion, biofiltration, composting), as well as subsurface injection of manure, are among the most encouraging options flagged in this section of the review. The importance of a multiple gas perspective is critical when assessing mitigation potentials, because most of the options reviewed show strong interactions among sources of greenhouse gas (GHG) emissions. The paper reviews current knowledge on potential pollution swapping, whereby the reduction of one GHG or emission source leads to unintended increases in another.

Six primiparous cannulated cows were assigned to six treatments in a 6 x 6 Latin square design to evaluate the effects of degree of fat saturation and amount and source of effective fiber on site of nutrient digestion and milk production. Cows were fed for ad libitum intake a control diet with no added fat or diets with 5% added fat from saturated tallow, tallow, or animal-vegetable fat; the diets with animal-vegetable fat had 40% forage, 40% forage plus 20% soyhulls, or 60% forage. Ruminal acetate:propionate was higher when soyhulls replaced forage NDF because of the higher digestibility of soyhulls in the rumen and total tract. Ruminal digestion of NDF was decreased as unsaturation of fat increased. True and apparent efficiencies of bacterial protein synthesis were increased as fat unsaturation increased, probably because of reduced recycling of microbial N in the rumen. The digestibility of fatty acids in the small intestine was higher in cows fed no fat than in those fed fat and was reduced as fat unsaturation decreased, primarily because of the saturated tallow. Dry matter intake was decreased 14% by increased fat unsaturation. All fat supplements depressed milk protein percentage. Production of 4% FCM tended to decrease linearly as unsaturation of fat increased. In diets with 5% added fat, a degree of saturation between saturated tallow and tallow (iodine values of 18 and 62) appears to be optimum for fatty acid digestibility and DMI.

Our objectives are to integrate current knowledge with a future perspective regarding how metagenomics can be used to integrate rumen microbiology and nutrition. Ruminal NH3-N concentration is a crude predictor of efficiency of dietary N conversion into microbial N, but as this concentration decreases below approximately 5 mg/dL (the value most often suggested to be the requirement for optimal microbial protein synthesis), blood urea N transfer into the rumen provides an increasing buffer against excessively low NH3-N concentrations, and the supply of amino N might become increasingly important to improve microbial function in dairy diets. Defaunation typically decreases NH3-N concentration, which should increase the efficiency of blood urea N and protein-derived NH3-N conversion into microbial protein in the rumen. Thus, we explain why more emphasis should be given toward characterization of protozoal interactions with proteolytic and deaminating bacterial populations. In contrast with research evaluating effects of protozoa on N metabolism, which has primarily been done with sheep and cattle with low dry matter intake, dairy cattle have greater intakes of readily available carbohydrate combined with increased ruminal passage rates. We argue that these conditions decrease protozoal biomass relative to bacterial biomass and increase the efficiency of protozoal growth, thus reducing the negative effects of bacterial predation compared with the beneficial effects that protozoa have on stabilizing the entire microbial ecosystem. A better understanding of mechanistic processes altering the production and uptake of amino N will help us to improve the overall conversion of dietary N into microbial protein and provide key information needed to further improve mechanistic models describing rumen function and evaluating dietary conditions that influence the efficiency of conversion of dietary N into milk protein.

Although many nonforage fiber sources have high extents of neutral detergent fiber (NDF) digestion, most have rates of digestion similar to or slower than the rates of forage NDF digestion. Rates of NDF digestion vary considerably among and within sources of by-products. Digestion kinetics also vary because of the technique used (in vitro versus in situ) and because of high amounts of dietary concentrate. Based on available data for passage rate and specific gravity measurements, rates of passage of nonforage fiber sources from the rumen of high producing cows appear to be faster than those of forages. Therefore, the potential to shift NDF digestion to the hindgut has been discussed. To account for variability in ruminal and total tract digestibility of NDF, multiple regression analysis was used to indicate that nonforage NDF percentage in the diet had about two-thirds the positive response on total tract NDF digestion that forage NDF percentage did. Although the loss of potentially digestible NDF may occur, DMI does not appear to decrease much until forage NDF is below 14 to 16% of dietary DM. Conversely, replacement of starch with nonforage NDF appears to increase digestibility of fiber, mostly in diets with high concentrations of nonfiber carbohydrates, apparently because of reduced negative associative effects. Increasing the concentration of total NDF above 35% also can decrease DMI with little improvement in NDF digestibility. Increased knowledge of the kinetics of digestion and the passage of various nonforage fiber sources used to replace forage or concentrate should increase the accuracy and precision of dynamic models, thereby increasing the flexibility and utility of nonforage fiber sources in dairy rations.

We have recently developed a real-time polymerase chain reaction (PCR) assay to quantify copies of the genes encoding protozoal 18S rRNA. The assay includes procedures for isolating and concentrating protozoal cells from the rumen for use as a standard to convert 18S rRNA gene copies to a biomass basis. The current objectives were to 1) determine the degree of reduction of bacterial contamination in the protozoal standard, 2) determine if protozoal standards derived from ruminal fluid are appropriate for predicting duodenal flows, and 3) evaluate the assay's determined values for protozoal N in the rumen and flowing to the duodenum compared with independent measurements. Our protozoal collection method reduced non-associated bacterial contamination by 33-fold, the contamination of which could otherwise significantly bias RNA (microbial marker) and N percentages of concentrated protozoal fractions. Based on denaturing gradient gel electrophoresis, the use of protozoal cells isolated from ruminal fluid appears appropriate for use in quantitative assays determining protozoal N flow postruminally. Using real-time PCR, protozoal N was determined to be 4.8 and 12.7% of the rumen microbial N pool and 5.9 and 11.9% of the duodenal flow of microbial N on diets containing low (16%) or high (21%) forage neutral detergent fiber, respectively, which were comparable with independent measures and expectations.