The objective of the present study was to investigate the effect of individual and combined use of dietary fat, nitrate and 3-nitrooxypropanol (3-NOP) on dairy cows' enteric methane (CH₄) emission and production performance. Twenty-four primiparous and 24 multiparous Danish Holstein cows (111 ± 44.6 d in milk; mean ± SD) were included in an incomplete 8 × 8 Latin square design with 6 21 d periods. Dietary treatments were organized in a 2 × 2 × 2 factorial arrangement aiming for 2 levels of FAT (30 or 63 g crude fat/kg of DM; LF or HF, respectively), 2 levels of NITRATE (0 or 10 g nitrate/kg of DM; UREA or NIT, respectively) and 2 levels of 3-NOP (0 or 80 mg/kg of DM; BLANK or NOP, respectively). Treatments were included in ad libitum fed partial mixed rations in bins that automatically measured feed intake and eating behavior. Additional concentrate was offered as bait in GreenFeed units used for measurement of gas emission. For total DM intake (DMI), a FAT × NITRATE interaction showed that DMI, across parities and levels of 3-NOP, was unaffected by separate fat supplementation, but reduced by nitrate with 4.6% and synergistically decreased (significant 2-way interaction) with 13.0% when fat and nitrate were combined. Additionally, 3-NOP decreased DMI by 13.4% and the combination of 3-NOP with fat and nitrate decreased DMI in an additive way (no significant 3-way interaction). The decreasing effects on DMI were more pronounced in multiparous cows than in primiparous cows. For treatments with largest reductions in DMI, eating behavior was altered toward more frequent, but smaller meals, a slower eating rate and increased attempts to visit unassigned feed bins. Energy-corrected milk (ECM) yield increased by 6.3% with fat supplementation, whereas ECM yield did not differ among diets including nitrate (FAT × NITRATE interaction). Cows supplemented with 3-NOP had 9.0% lower ECM yield than cows fed no 3-NOP. Based on 3 2-way interactions including FAT, NITRATE and 3-NOP, the combined use of the additives resulted in antagonistic effects on CH₄ reduction. A 6 to 7% reduction in CH₄ yield (CH₄/kg of DMI) could be ascribed to the effect of fat, a 12 to 13% reduction could be ascribed to the effect of nitrate and an 18 to 23% reduction could be ascribed to the effect of 3-NOP. Hence, no combinations of additives resulted in CH₄ yield-reductions that were greater than what was obtained by separate supplementation of the most potent additive within the combination. The CH₄ yield reduction potential of additives was similar between parities. Increased apparent total-tract digestibility of organic matter (OM) in cows fed combinations including nitrate and/or 3-NOP was a result of a NITRATE × 3-NOP interaction. Apparent total-tract digestibility of OM was also increased by fat supplementation. These increases reflected observed decreases in DMI. In conclusion, combined use of fat, nitrate and 3-NOP in all combinations did not result in CH₄ reductions that were greater than separate supplementation of the most potent additive within the combination (3-NOP > NITRATE > FAT). Additionally, separate supplementation of some additives and combined use of all additives reduced DMI.

Due to climate change, periods of drought might be longer and occur more frequently, which challenges roughage production and requires changed feeding of dairy cattle by increasing the grain content of the diet. This study investigated the effect of diets with concentrate proportions up to 91% of dry matter on dry matter intake (DMI), milk production, enteric methane emission, rumen fermentation, rumen bacterial community structure, nutrient digestibility, and feeding behavior of Holstein and Jersey dairy cows. Twelve Danish Holstein and 12 Danish Jersey cows were fed ad libitum with one of 3 total mixed rations differing in concentrate proportion in a continuous design with staggered approach over 19 to 29 d. Dietary concentrate proportions were 49% (C49), 70% (C70), and 91% (C91) on dry matter basis, and were based on increasing proportions of chopped barley straw, dried beet pulp, barley, NaOH-treated wheat, dried distillers grain, and rapeseed cake at the expense of grass/clover silage, corn silage and soybean meal. Cows were adapted to the diets over a 12- to 19-d period, before rumination activity was measured over 3 d. Subsequently, spot samples of feces were collected for digestibility determination over 2 d, and gas exchange was measured on the last 3 d of the experimental period. Shortly after chamber stay, rumen liquid was collected using an oro-ruminal device. Dry matter intake was higher for Holstein than Jersey. Methane emissions (all expressions) were affected by the interaction between breed and diet. Methane per kilogram of DMI was lowered by 18 and 48% for Holstein fed C70 and C91, respectively, compared with C49, whereas this was 17 and 22% respectively for Jersey. Rumen propionate molar proportion increased more, rumen bacterial community was less diverse, and rumination time and rumination chews relative to DMI reduced less for Holstein than for Jersey cows with increasing concentrate level. In conclusion, Holstein dairy cows responded stronger to increased dietary concentrate level regarding methane mitigation, changes in rumen VFA profile, and effect on the rumen bacterial community structure than Jersey cows, whereas Jersey cows responded stronger with regard to rumination time and rumination chews (per kilogram of DMI and per kilogram of neutral detergent fiber intake) than Holstein cows. Thus, diets high in concentrates are a less effective methane mitigation strategy for Jersey than for Holstein.

Essential oils (EO) from oregano may have antimicrobial properties, potentially representing a methane mitigation strategy suitable for organic production. This study aimed to (1) examine the potential of oregano in lowering enteric methane production of dairy cows fed differing levels of dried oregano (Origanum vulgare ssp. hirtum) plant material containing high levels of EO; (2) determine whether differing levels of dried oregano plant material of another subspecies (Origanum vulgare ssp. vulgare) with naturally low levels of EO in feed affected enteric methane production; and (3) evaluate the effect of various levels of the 2 oregano subspecies (containing high or low levels of EO) in feed on rumen fermentation, nutrient digestibility, and milk fatty acids. Each experiment had a 4 × 4 Latin square design using 4 lactating Danish Holstein dairy cows that had rumen, duodenal, and ileal cannulas and were fed 4 different levels of oregano. Experiment 1 used low EO oregano [0.12% EO of oregano dry matter (DM)] and evaluated a control (C) diet with no oregano and 3 oregano diets with 18 (low; L), 36 (medium; M), and 53 g of oregano DM/kg of dietary DM (high; H). Experiment 2 used high EO oregano (4.21% EO of oregano DM) with 0, 7, 14, and 21 g of oregano DM/kg of dietary DM for C, L, M, and H, respectively. Oregano was added to the diets by substituting grass/clover silage on a DM basis. Low or high EO oregano in feed did not affect dry matter intake (DMI) or methane production (grams per day, grams per kilogram of DMI, grams per kilogram of energy-corrected milk, and percentage of gross energy intake). Rumen fermentation was slightly affected by diet in experiment 1, but was not affected by diet in experiment 2. In both experiments, the apparent total-tract digestibility of DM, organic matter, and neutral detergent fiber decreased linearly and cubically (a cubic response was not observed for neutral detergent fiber) with increasing dietary oregano content, while milk fatty acids were slightly affected. In conclusion, dried oregano plant material with either high or low levels of EO did not lower the methane production of dairy cows over 4 consecutive days, and no substantial effects were observed on rumen fermentation or nutrient digestibility. This conclusion regarding methane production is in contrast with literature and requires further study.

Manure nitrogen (N) from cattle contributes to nitrous oxide and ammonia emissions and nitrate leaching. Measurement of manure N outputs on dairy farms is laborious, expensive, and impractical at large scales; therefore, models are needed to predict N excreted in urine and feces. Building robust prediction models requires extensive data from animals under different management systems worldwide. Thus, the study objectives were (1) to collate an international database of N excretion in feces and urine based on individual lactating dairy cow data from different continents; (2) to determine the suitability of key variables for predicting fecal, urinary, and total manure N excretion; and (3) to develop robust and reliable N excretion prediction models based on individual data from lactating dairy cows consuming various diets. A raw data set was created based on 5,483 individual cow observations, with 5,420 fecal N excretion and 3,621 urine N excretion measurements collected from 162 in vivo experiments conducted by 22 research institutes mostly located in Europe (n = 14) and North America (n = 5). A sequential approach was taken in developing models with increasing complexity by incrementally adding variables that had a significant individual effect on fecal, urinary, or total manure N excretion. Nitrogen excretion was predicted by fitting linear mixed models including experiment as a random effect. Simple models requiring dry matter intake (DMI) or N intake performed better for predicting fecal N excretion than simple models using diet nutrient composition or milk performance parameters. Simple models based on N intake performed better for urinary and total manure N excretion than those based on DMI, but simple models using milk urea N (MUN) and N intake performed even better for urinary N excretion. The full model predicting fecal N excretion had similar performance to simple models based on DMI but included several independent variables (DMI, diet crude protein content, diet neutral detergent fiber content, milk protein), depending on the location, and had root mean square prediction errors as a fraction of the observed mean values of 19.1% for intercontinental, 19.8% for European, and 17.7% for North American data sets. Complex total manure N excretion models based on N intake and MUN led to prediction errors of about 13.0% to 14.0%, which were comparable to models based on N intake alone. Intercepts and slopes of variables in optimal prediction equations developed on intercontinental, European, and North American bases differed from each other, and therefore region-specific models are preferred to predict N excretion. In conclusion, region-specific models that include information on DMI or N intake and MUN are required for good prediction of fecal, urinary, and total manure N excretion. In absence of intake data, region-specific complex equations using easily and routinely measured variables to predict fecal, urinary, or total manure N excretion may be used, but these equations have lower performance than equations based on intake.

Many essential oils and their terpene constituents display antimicrobial properties, which may affect rumen metabolism and influence milk production parameters. Many of these compounds also have distinct flavors and aromas that may make their way into the milk, altering its sensory properties. Essential oils from caraway (Carum carvi) seeds and oregano (Origanum vulgare) plants were included in dairy cow diets to study the effects on terpene composition and sensory properties of the produced milk, as well as feed consumption, production levels of milk, and methane emissions. Two levels of essential oils, 0.2 and 1.0g of oil/kg of dry matter, were added to the feed of lactating cows for 24d. No effects on feed consumption, milk production, and methane emissions were observed. The amount and composition of volatile terpenes were altered in the produced milk based on the terpene content of the essential oils used, with the total amount of terpenes increasing when essential oils were added to the diet. Sensory properties of the produced milk were altered as well, and milk samples from animals receiving essential oil treatment were perceived as having a fresher aroma and lower stored aroma and flavor. The levels of essential oils used in this study mimic realistic levels of essential oils in herbs from feed, but were too low to affect milk production and methane emissions, and their inclusion in the animal diet did not adversely affect milk flavor.

Devices that record behavior automatically have made it possible to accurately measure the lying and eating behavior of large numbers of dairy cows. During lactation, weight, feed intake, and production of cows change; however, longitudinal studies of how the behavior of dairy cows is correlated with production traits during lactation are limited. This study describes changes in duration of lying and eating behavior throughout lactation and how these variables are related to changes in milk yield, live weight, and feed intake in lactating primi- and multiparous Holstein and Jersey cows. Data were from 255 cow lactations (43 primi- and 80 multiparous Jersey cows, and 56 primi- and 76 multiparous Holstein cows) from 5 to 200 d in milk. Leg-mounted tags were used to record lying time and steps; ad libitum feed intake (of a partial mixed ration) variables were recorded from feed bins on weight cells; and milk yield and live weight were recorded during automatic milking, all on a daily basis. The lactation trajectory was split into 4 segments. Data were analyzed using mixed effects linear models. Holstein cows spent more time lying and eating than Jersey cows, whereas Jersey cows had a greater number of steps (25-37%). First-lactation cows spent less time eating and had more steps than older cows. Average daily lying time was approximately 1 h longer during February than the shortest lying time, which was observed in August. Both Holstein and Jersey multiparous cows had longer lying times than cows in first parity after parturition; however, the lying time of multiparous cows decreased, whereas that of primiparous cows increased in the beginning of lactation. Later in lactation, older cows tended to increase duration of lying more than younger cows did. The daily change in behavior (lying, eating, and steps) and milk yield, live weight, and dry matter intake, characterized as slopes in the lactation period for each cow, were not strongly correlated. However, we found a moderate correlation between changes in milk yield and dry matter intake, and between changes in eating time and rate of eating. An increase in eating rate in multiparous Holstein cows was correlated with increasing lying time. In conclusion, the use of automated behavior recording enabled thorough investigations of relationships between a range of behavior traits and frequently recorded production traits, and revealed that patterns of change during lactation are strongly affected by breed and parity.