Potential of 2 Northern European brown seaweeds (Fucus serratus and Fucus vesiculosus) as enteric methane inhibitors in dairy cows.

The 2 brown seaweeds, Fucus serratus and Fucus vesiculosus, have demonstrated anti-methanogenic properties in vitro with reductions in CH4 production ranging from 53 to 63%. This study aimed to investigate the effects of the 2 Fucus seaweeds on enteric CH4 emissions, DMI, ECM, and nutrient digestibility when fed to dairy cows. The experiment was conducted using 4 multi-cannulated lactating Danish Holstein dairy cows, which over 3 experimental periods received either: 1) basal diet (CON; diet without any seaweed), 2) basal diet diluted with 4% (DM basis) Fucus serratus (SER), or 3) basal diet diluted with 4% (DM basis) Fucus vesiculosus (VES); resulting in one complete 3 × 3 Latin square and one incomplete 3 × 3 Latin square. Each period lasted 21 d and consisted of 14 d of adaptation, followed by 3 d of digesta sampling, and 4 d of gas exchange measurements using respiration chambers. Milk yield and feed intake were recorded daily. Blood samples were collected on d 15 and 17. All parameters were statistically analyzed using a mixed procedure of R. Opposite to what we had expected, neither of the 2 Fucus seaweeds reduced CH4 emissions from the dairy cows as daily CH4 production was significantly higher for both Fucus treatments compared with CON. Additionally, CH4 yield (g CH4/kg DMI) and intensity (g CH4/kg ECM) were significantly higher for SER compared with CON. Milk yield, DMI, and total-tract digestibility were unaffected by the treatments; however, SER resulted in lower milk protein yield (kg/d) and lower milk and blood plasma urea concentrations compared with CON. In conclusion, neither Fucus serratus (SER) nor Fucus vesiculosus (VES) showed potential as methane-mitigating feed additives when fed to dairy cows at an inclusion level of 4% of DM. The inclusion of the 2 brown seaweeds had no effects on DMI, milk yield, or total-tract digestibility.

Predicting CO2 production of lactating dairy cows from animal, dietary, and production traits using an international dataset.

Automated measurements of the ratio of concentrations of methane and carbon dioxide, [CH4]:[CO2], in breath from individual animals (the so-called "sniffer technique") and estimated CO2 production can be used to estimate CH4 production, provided that CO2 production can be reliably calculated. This would allow CH4 production from individual cows to be estimated in large cohorts of cows, whereby ranking of cows according to their CH4 production might become possible and their values could be used for breeding of low CH4-emitting animals. Estimates of CO2 production are typically based on predictions of heat production, which can be calculated from body weight (BW), energy-corrected milk yield, and days of pregnancy. The objectives of the present study were to develop predictions of CO2 production directly from milk production, dietary, and animal variables, and furthermore to develop different models to be used for different scenarios, depending on available data. An international dataset with 2,244 records from individual lactating cows including CO2 production and associated traits, as dry matter intake (DMI), diet composition, BW, milk production and composition, days in milk, and days pregnant, was compiled to constitute the training dataset. Research location and experiment nested within research location were included as random intercepts. The method of CO2 production measurement (respiration chamber [RC] or GreenFeed [GF]) was confounded with research location, and therefore excluded from the model. In total, 3 models were developed based on the current training dataset: model 1 ("best model"), where all significant traits were included; model 2 ("on-farm model"), where DMI was excluded; and model 3 ("reduced on-farm model"), where both DMI and BW were excluded. Evaluation on test dat sets with either RC data (n = 103), GF data without additives (n = 478), or GF data only including observations where nitrate, 3-nitrooxypropanol (3-NOP), or a combination of nitrate and 3-NOP were fed to the cows (GF+: n = 295), showed good precision of the 3 models, illustrated by low slope bias both in absolute values (-0.22 to 0.097) and in percentage (0.049 to 4.89) of mean square error (MSE). However, the mean bias (MB) indicated systematic overprediction and underprediction of CO2 production when the models were evaluated on the GF and the RC test datasets, respectively. To address this bias, the 3 models were evaluated on a modified test dataset, where the CO2 production (g/d) was adjusted by subtracting (where measurements were obtained by RC) or adding absolute MB (where measurements were obtained by GF) from evaluation of the specific model on RC, GF, and GF+ test datasets. With this modification, the absolute values of MB and MB as percentage of MSE became negligible. In conclusion, the 3 models were precise in predicting CO2 production from lactating dairy cows.

Effects of dietary inclusion of 3 Nordic brown macroalgae on enteric methane emission and productivity of dairy cows.

Macroalgae are receiving increased attention as antimethanogenic feed additives for cattle, but most in vivo studies are limited to investigating effects of the red macroalgae Asparagopsis spp. Hence, this study aimed to investigate the CH4 mitigating potential of 3 brown macroalgae from the Northern Hemisphere when fed to dairy cows, and to study the effects on feed intake, milk production, feed digestibility, and animal health indicators. The experiment was conducted as a 4 × 4 Latin square design using 4 lactating rumen, duodenal, and ileal cannulated Danish Holstein dairy cows. The cows were fed a total mixed ration (TMR) without any macroalgae or the same TMR diluted with, on a dry matter basis, either 4% ensiled Saccharina latissima, 4% Ascophyllum nodosum (NOD), or 2% Sargassum muticum (MUT). Each period consisted of 14 d of adaptation, 3 d of digesta and blood sampling, and 4 d of gas exchange measurements using respiration chambers. Milk yield and dry matter intake (DMI) were recorded daily. Blood was sampled on d 13 and 16 and analyzed for health status indicators. None of the 3 species affected the CH4 emission. Moreover, milk yield and DMI were also unaffected. Total-tract digestibility of crude protein was significantly lower for NOD compared with other diets, and additionally, the NOD diet also tended to reduce total-tract digestibility of neutral detergent fiber compared with MUT. Blood biomarkers did not indicate negative effects of the dietary inclusion of macroalgae on cow health. In conclusion, none of the 3 brown macroalgae reduced CH4 emission and did not affect DMI and milk production of dairy cows, whereas negative effects on the digestibility of nutrients were observed when A. nodosum was added. None of the diets would be allowed to be fed in commercial dairy herds due to high contents of iodine, cadmium, and arsenic.

Fiber digestibility and protein value of pulp silage for lactating dairy cows: Effects of wet fractionation by screw pressing of perennial ryegrass.

The aim of the study was to investigate the effects of substituting silage of chopped grass with pulp silage of grass fractionated once or twice in a biorefinery using a screw press on fiber kinetics, protein value, and production of CH4 in dairy cows. Six lactating multiparous Holstein cows in mid-lactation (176 ± 93 d in milk), cannulated in the rumen, duodenum, and ileum, were used in an incomplete 6 × 4 Latin square design with a 2 × 3 factorial arrangement of treatments. Perennial ryegrass was harvested in third regrowth from the same field at early and late developmental stage (35 and 44 d of regrowth, respectively) and subjected to 1 of 3 types of processing within each developmental stage. Grass was either harvested for normal silage making (mowed, wilted, chopped, and ensiled), or harvested fresh and fractionated using a screw press. Half of the pulp from the first fractionation was ensiled, whereas the other half of the pulp was rehydrated, fractionated a second time, and pulp hereof was ensiled. The grass and pulp silages were used with concentrates (65:35 forage to concentrate ratio) to make total mixed rations (TMR) based on either silage of chopped grass (GS), pulp silage of grass fractionated once (1×P), or pulp silage of grass fractionated twice (2×P), harvested either at early (E) or late (L) developmental stage resulting in 6 different TMR treatments (EGS, E1×P, E2×P, LGS, L1×P, L2×P). The TMR were fed for ad libitum intake and samples of intestinal digesta and feces were collected for determination of digestibility. The effect of processing on ash-free neutral detergent fiber (aNDFom) concentration in silages depended on developmental stage, but showed that within each developmental stage, pulp silage of grass fractionated twice had higher aNDFom concentration than pulp silage of grass fractionated once and silage of chopped grass. The 2×P resulted in lower (14.9 ± 0.55 vs. 17.5 ± 0.54 kg/d) dry matter intake (DMI) compared with GS. The effects of processing and developmental stage interacted such that apparent total-tract aNDFom digestibility was higher (784 ± 13 vs. 715 ± 13 g/kg) for L2×P compared with LGS, whereas no difference was found between E2×P and EGS. Moreover, the protein value was higher (106 ± 5 vs. 92 ± 5 g AA digested in the small intestine/kg of DMI) for 2×P compared with GS. Unexpectedly, processing had no effect on fractional rate of digestion of digestible aNDFom or CH4 yield (L/kg of DMI), whereas feeding forages harvested at early compared with late developmental stage resulted in lower CH4 yield. Feeding pulp silage of grass fractionated once generally yielded results intermediate to cows fed silage of chopped grass and pulp silage of grass fractionated twice. This study showed that pulp silage of fractionated grass could serve as feed for dairy cows because the fiber digestibility and protein value improved, but further research investigating effects of physical processing of forage on fiber kinetics is required.

Methane emission, nutrient digestibility, and rumen microbiota in Holstein heifers fed 14 different grass or clover silages as the sole feed.

This experiment investigated the variation in enteric methane production and associated gas exchange parameters, nutrient digestibility, rumen fermentation, and rumen microbiome when a range of silages based on different forage types (grass or clover), and different species within the 2 types, were fed as the sole feed to heifers. Three grass species (perennial ryegrass, festulolium, and tall fescue) and 2 clover species (red clover and white clover) were included. Perennial ryegrass was harvested at 2 maturity stages in the primary growth, white clover was harvested once in the primary growth, and 4 cuts of festulolium and tall fescue and 3 cuts of red clover were harvested during the growing season, giving 14 different silage batches in total. Sixteen Holstein heifers 16 to 21 mo old and 2 to 5 mo in pregnancy were fed the silages ad libitum as the sole feed in an incomplete crossover design. Each silage was fed to 4 heifers, except for the 2 perennial ryegrass silages, which were fed to 8 heifers; in total 64 observations. The CH4 production was measured for 3 d in respiration chambers. Heifers fed clover silages had higher dry matter intake (DMI) compared with heifers fed grass silages, and heifers fed tall fescue silages had the numerically the lowest DMI. Compared with grass silages, feeding clover silages led to higher crude protein digestibility but lower neutral detergent fiber (NDF) digestibility. Rumen pH was higher in heifers fed clover silages compared with those fed grass silages. Based on composition analysis, the rumen microbiota of the heifers clustered clearly according to forage type and species. More specifically, 7 of the 34 dominating rumen bacterial genus-level groups showed higher relative abundances for the clover silages, whereas 7 genus-level groups showed higher abundances for the grass silages. Methane yield was higher for heifers fed grass silages than for those fed clover silages when methane production was related to dry matter and digestible organic matter intake, whereas the opposite was seen when related to NDF digestion. The gross energy lost as methane (CH4 conversion factor, %) reduced from 7.5% to 6.7%, equivalent to an 11% reduction. The present study gives the outlines for choosing the optimal forage type and forage species with respect to nutrient digestibility and enteric methane emission in ruminants.

Effects of particle size and toasting of fava beans and forage source on nutrient digestibility, ruminal fermentation, and metabolizable protein supply in dairy cows.

The objective of this study was to investigate the effects of heat treatment (toasting) and particle size alterations (grinding; rolling) on nutrient utilization, ruminal fermentation, and supply of metabolizable protein (MP), and to study the interaction between processing conditions of fava beans and forage type. Six Danish Holstein dairy cows fitted with ruminal, duodenal, and ileal cannulas were used in a 6 × 4 incomplete Latin square design with 4 periods of 21-d duration. Cows were fed ad libitum with 6 experimental diets: diets high in either grass-clover silage or corn silage were combined with ground untoasted, ground toasted, or rolled untoasted fava beans. Samples of ruminal fluid, digesta from duodenum and ileum, and feces were collected, and nutrient digestibility was estimated using Cr2O3 and TiO2 as flow markers. Diets high in corn silage resulted in higher ruminal pH and higher proportion of propionate in ruminal volatile fatty acids compared with diets high in grass-clover silage. Diets high in corn silage resulted in higher apparent total-tract digestibility of crude protein and starch but lower apparent ruminal and total-tract digestibility of neutral detergent fiber compared with diets high in grass-clover silage. Rolling of fava beans decreased the in situ small intestinal disappearance of rumen-undegradable protein corrected for particle losses. Compared with grinding, rolling of fava beans reduced apparent ruminal digestibility of starch, true ruminal digestibility of organic matter, crude protein, and AA, and small intestinal digestibility of AA and starch. Grinding of fava beans increased apparent ruminal digestibility of neutral detergent fiber and reduced the proportion of propionate in ruminal volatile fatty acids compared with rolling of fava beans. In addition, rolling of fava beans had no effect on MP supply. Toasting of fava beans had no effect on in vivo nutrient digestibility except for an interaction with forage source on apparent ruminal dry matter and organic matter digestibility. Toasting of fava beans did not affect small intestinal digestion of individual and total AA, and therefore failed to increase MP supply. In conclusion, neither replacing grass-clover silage with corn silage, nor toasting nor rolling of fava beans had an effect on supply of MP.

Feeding up to 91% concentrate to Holstein and Jersey dairy cows: Effects on enteric methane emission, rumen fermentation and bacterial community, digestibility, production, and feeding behavior.

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.

Effects on feed intake, milk production, and methane emission in dairy cows fed silage or fresh grass with concentrate or fresh grass harvested at early or late maturity stage without concentrate.

The objective of the study was to quantify the effects on dry matter intake (DMI), nutrient digestibility, gas exchange, milk production, and milk quality in dairy cows fed fresh grass harvested at different maturity stages. Sixteen Danish Holstein cows in mid-lactation were divided into 4 blocks and used in 4 incomplete 4 × 2 Latin squares with 2 periods of 21 d. The cows received 1 of 4 treatments in each period, resulting in 8 cows per treatment, as follows: grass-clover silage supplemented with 6 kg/d concentrate pellets (SILc), fresh grass harvested at late maturity stage supplemented with 6 kg/d concentrate pellets (LATc), fresh grass harvested at late maturity stage (LAT), and fresh grass harvested at early maturity stage (ERL). The cows were housed in tiestalls and milked twice daily. The cows had ad libitum access to the forage, and concentrate pellets were divided into equal amounts and fed separately in the morning and afternoon. Fecal samples were collected to determine apparent total-tract digestibility, and samples of rumen fluid were collected for determination of short chain fatty acid composition. Halters were used for measuring eating and rumination time. Gas exchange was measured in open-circuit respiration chambers. Total DMI was higher in LATc and ERL (16.9 ± 0.45 and 15.5 ± 0.39 kg/d, respectively) compared with LAT (14.1 ± 0.42 kg/d). Relative to SILc, cows fed fresh grass experienced a convex pattern in DMI during the experiment. The changes in DMI were related to changes in leaf to stem ratio, fiber concentration, and organic matter digestibility determined in vitro in samples of the fresh grass harvested throughout the experiment. The apparent total-tract digestibility of organic matter was higher in SILc and LAT compared with LATc. Methane yield was lower for LATc compared with LAT (19.5 ± 0.61 vs. 22.6 ± 0.55 g of CH4/kg of DMI), and was not different between LAT and ERL. Compared with LAT, milk yield was higher for ERL (21.1 ± 1.14 vs. 23.4 ± 1.11 kg/d) and energy-corrected milk (ECM) yield was higher for LATc (21.5 ± 0.99 vs. 25.3 ± 1.03 kg/d). We detected no differences in milk or ECM yield between SILc and LATc. Milk protein yield was higher and milk fat concentration was lower in LATc compared with LAT. The fatty acid percentages of ∑C4-C14:1 and ∑C16 in milk were higher for SILc compared with LATc, signifying pronounced de novo synthesis. The n-6:n-3 ratio in milk fatty acids was lower for SILc and LAT compared with LATc, indicating improved nutritional quality for SILc and LAT. However, retinol concentration in milk was lower in SILc compared with all other treatments. The study implies that feeding silage instead of fresh grass has no effect on DMI, ECM yield, or CH4 yield, and that concentrate supplementation can increase milk production, affects milk quality, and reduces the effect on climate, whereas feeding less mature grass increases DMI and milk yield, but has no effect on CH4 yield.

Enteric methane emission and digestion in dairy cows fed wheat or molasses.

The aim of this experiment was to measure enteric methane (CH4) emission and its relation with rumen digestion in dairy cows fed diets rich in 1 of the 2 carbohydrate sources, starch or sugar. The rations were based on late first-cut grass-clover silage supplemented with wheat (Wh), NaOH-treated wheat (Wh+NaOH), sugar beet molasses (Mo), or sugar beet molasses with addition of sodium bicarbonate (Mo+Bic). Wheat and molasses made up 35% of dry matter in the 2 diets with molasses and wheat, respectively. Four cows fitted with ruminal, duodenal, and ileal canulae were used in a 4 × 4 Latin square design. Nutrient digestibility was measured using chromium oxide and titanium oxide as flow markers, and emissions of CH4 and hydrogen were measured via open-circuit indirect calorimetry on 4 consecutive days. Data were analyzed using PROC MIXED of SAS (version 9.4; SAS Institute Inc., Cary, NC) with treatment and period as fixed effects and cow as random effect. Furthermore, orthogonal contrasts were calculated. The cows produced 32.5, 33.6, 36.2, and 35.1 L of CH4/kg of dry matter intake (DMI) on diets Wh, Wh+NaOH, Mo, and Mo+Bic, respectively. The emission of CH4 per day, per kilogram of DMI, and per kilogram of energy-corrected milk as well as daily hydrogen emission were higher on the Mo diet compared with the Wh diet. With the present inclusion of wheat and molasses in the diet, no effects of NaOH treatment of wheat or of sodium bicarbonate supplementation to the Mo diet could be demonstrated on CH4 emission expressed per kilogram of DMI or per kilogram of energy-corrected milk. The duodenal flow of starch was higher when wheat was treated with NaOH. Under the conditions in the present experiment, ruminal NDF digestibility was not affected by carbohydrate source, NaOH treatment of wheat, or bicarbonate supplementation. Total volatile fatty acid concentration in the rumen and the proportions of acetate and propionate were not affected by carbohydrate source, NaOH treatment of wheat, or bicarbonate supplementation. Likewise, we could not show any influence of diet on microbial protein synthesis or efficiency of microbial protein synthesis expressed as grams of microbial protein synthesis per kilogram of true rumen-digested organic matter. We concluded that CH4 emission was increased when wheat was replaced by molasses, whereas no effect of manipulating rumen fermentation by NaOH treatment of wheat or addition of bicarbonate to molasses could be found with a level of approximately 25% of dry matter from starch and sugar, respectively.

Short communication: Variation in feed efficiency hampers use of carbon dioxide as a tracer gas in measuring methane emissions in on-farm conditions.

Breeding cows for low CH4 emissions requires that the trait is variable and that it can be recorded with low cost from an adequate number of individuals and with high precision, but not necessarily with high accuracy if the trait is measured with high repeatability. The CH4:CO2 ratio in expired breath is a trait often used as a tracer with the production of CO2 predicted from body weight (BW), energy-corrected milk yield, and days of pregnancy. This approach assumes that efficiency of energy utilization for maintenance and production is constant. Data (307 cow-period observations) from 2 locations using the same setup for measuring CH4 and CO2 in respiration chambers were compiled, and observed production of CH4 and CO2 was compared with the equivalent predicted production using 2 different approaches. Carbon dioxide production was predicted using a previously reported model based on metabolic BW and energy-corrected milk production and a currently developed model based on energy requirements and the relationship between observed CO2 and heat production (models 1 and 2, respectively). Animals used were categorized (low, medium, and high efficiency) according to (1) residual feed intake and (2) residual milk production. Model 1 underestimated CH4 production by 15%, whereas model 2 overestimated CH4 by 1.4% for the whole database. Model 1 underestimated CO2 production by 2.8 and 0.9 kg/d for low- and high-efficiency cows, respectively, whereas model 2 underestimated CO2 production by 0.9 kg/d for low-efficient animals but overestimated it by 1.2 kg/d for high-efficiency cows. Efficient cows produce less heat, and consequently CO2, per unit of metabolic body weight and energy-corrected milk than inefficient cows, challenging the use of CO2 as a tracer gas. Because of biased estimates of CO2 production, the models overestimated CH4 production of high-efficiency cows by, on average, 17% relative to low-efficiency cows, respectively. Selecting low CH4-emitting cows using a CO2 tracer method can therefore favor inefficient cows over efficient cows.

Between-cow variation in the components of feed efficiency.

A meta-analysis based on an individual-cow data set was conducted to investigate between-cow variations in the components and measurements of feed efficiency (FE) and to explore the associations among these components. Data were taken from 31 chamber studies, consisting of a total of 841 cow/period observations. The experimental diets were based on grass or corn silages, fresh grass, or a mixture of fresh grass and straw, with cereal grains or by-products as energy supplements, and soybean or canola meal as protein supplements. The average forage-to-concentrate ratio across all diets on a dry matter basis was 56:44. Variance component and repeatability estimates of FE measurements and components were determined using diet, period, and cow within experiment as random effects in mixed procedures of SAS (SAS Institute Inc., Cary, NC). The between-cow coefficient of variation (CV) in gross energy intake (GE; CV = 0.10) and milk energy (El) output as a proportion of GE (El/GE; CV = 0.084) were the largest among all component traits. Similarly, the highest repeatability estimates (≥0.50) were observed for these 2 components. However, the between-cow CV in digestibility (DE/GE), metabolizability [metabolizable energy (ME)/GE], methane yield (CH4E/GE), proportional urinary energy output (UE/GE), and heat production (HP/GE), as well as the efficiency of ME use for lactation (kl), were rather small. The least repeatable component of FE was UE/GE. For FE measurements, the between-cow CV in residual energy-corrected milk (RECM) was larger than for residual feed intake (RFI), suggesting a greater possibility for genetic gain in RECM than in RFI. A high DE/GE was associated with increased CH4E/GE (r = 0.24), HP/GE (r = 0.12), ME/GE (r = 0. 91), energy balance as a proportion of GE (EB/GE; r = 0.35), and kl (r = 0.10). However, no correlation between DE/GE and GE intake or UE/GE was observed. Increased proportional milk energy adjusted to zero energy balance (El(0)/GE) was associated with increases in DE/GE, ME/GE, EB/GE, and kl but decreases in UE/GE, CH4E/GE, and HP/GE, with no effect on GE intake. In conclusion, several mechanisms are involved in the observed differences in FE among dairy cows, and reducing CH4E yield (CH4E/GE) may inadvertently result in reduced GE digestibility. However, the selection of dairy cows with improved energy utilization efficiencies offers an effective approach to lower enteric CH4 emissions.

Effect of dried oregano (Origanum vulgare L.) plant material in feed on methane production, rumen fermentation, nutrient digestibility, and milk fatty acid composition in dairy cows.

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.

Methane production, rumen fermentation, and diet digestibility of Holstein and Jersey dairy cows being divergent in residual feed intake and fed at 2 forage-to-concentrate ratios.

Improving feed efficiency of dairy cows through breeding is expected to reduce enteric methane production per unit of milk produced. This study examined the effect of 2 forage-to-concentrate ratios on methane production, rumen fermentation, and nutrient digestibility in Holstein and Jersey dairy cows divergent in residual feed intake (RFI). Before experimental onset, RFI was estimated using a random regression model on phenotypic herd data. Ten lactating Holstein and 10 lactating Jersey cows were extracted from the herd and allocated to a high or low pre-experimental RFI group of 5 animals each within breed. Cows were fed ad libitum with total mixed rations either low (LC) or high (HC) in concentrates during 3 periods in a crossover design with a back-cross and staggered approach. Forage-to-concentrate ratio was 68:32 for LC and 39:61 for HC. Cows adapted to the diets in 12 to 24 d and feces were subsequently collected on 2 d. Afterward, gas exchange was measured in respiration chambers and rumen liquid was collected once after cows exited the chambers. Pre-experimental RFI was included in the statistical analysis as a class (low and high RFI) or continuous variable. Methane per kilogram of dry matter intake (DMI) was lower for Holsteins than Jerseys and the response to increased concentrate level was more pronounced for Holsteins than Jerseys (27.2 vs.13.8%); a similar pattern was found for the acetate:propionate ratio. However, methane production per kilogram of energy-corrected milk (ECM) was unaffected by breed. Further, total-tract digestibility of neutral detergent fiber was higher for Jerseys than Holsteins. For RFI as a class variable, DMI, methane production regardless of the expression, and digestibility were unaffected by RFI. For RFI as a continuous variable, DMI was lower and methane per kilogram of DMI was higher for cows with negative (efficient) than positive (inefficient) RFI values, and neutral detergent fiber digestibility was higher for Holsteins with negative than positive RFI values, but not for Jerseys. Daily methane production and methane per kilogram of ECM were unaffected by RFI. In conclusion, methane per kilogram of DMI of Jerseys was lowered to a smaller extent in response to the HC diet than of Holsteins. When pre-experimental RFI was used as a continuous variable, higher methane per kilogram of DMI was found for cows with negative RFI than positive RFI values, but not for methane per kilogram of ECM. These findings call for validation in larger studies.

Symposium review: Uncertainties in enteric methane inventories, measurement techniques, and prediction models.

Ruminant production systems are important contributors to anthropogenic methane (CH4) emissions, but there are large uncertainties in national and global livestock CH4 inventories. Sources of uncertainty in enteric CH4 emissions include animal inventories, feed dry matter intake (DMI), ingredient and chemical composition of the diets, and CH4 emission factors. There is also significant uncertainty associated with enteric CH4 measurements. The most widely used techniques are respiration chambers, the sulfur hexafluoride (SF6) tracer technique, and the automated head-chamber system (GreenFeed; C-Lock Inc., Rapid City, SD). All 3 methods have been successfully used in a large number of experiments with dairy or beef cattle in various environmental conditions, although studies that compare techniques have reported inconsistent results. Although different types of models have been developed to predict enteric CH4 emissions, relatively simple empirical (statistical) models have been commonly used for inventory purposes because of their broad applicability and ease of use compared with more detailed empirical and process-based mechanistic models. However, extant empirical models used to predict enteric CH4 emissions suffer from narrow spatial focus, limited observations, and limitations of the statistical technique used. Therefore, prediction models must be developed from robust data sets that can only be generated through collaboration of scientists across the world. To achieve high prediction accuracy, these data sets should encompass a wide range of diets and production systems within regions and globally. Overall, enteric CH4 prediction models are based on various animal or feed characteristic inputs but are dominated by DMI in one form or another. As a result, accurate prediction of DMI is essential for accurate prediction of livestock CH4 emissions. Analysis of a large data set of individual dairy cattle data showed that simplified enteric CH4 prediction models based on DMI alone or DMI and limited feed- or animal-related inputs can predict average CH4 emission with a similar accuracy to more complex empirical models. These simplified models can be reliably used for emission inventory purposes.

Effect of dietary nitrate level on enteric methane production, hydrogen emission, rumen fermentation, and nutrient digestibility in dairy cows.

Nitrate may lower methane production in ruminants by competing with methanogenesis for available hydrogen in the rumen. This study evaluated the effect of 4 levels of dietary nitrate addition on enteric methane production, hydrogen emission, feed intake, rumen fermentation, nutrient digestibility, microbial protein synthesis, and blood methemoglobin. In a 4×4 Latin square design 4 lactating Danish Holstein dairy cows fitted with rumen, duodenal, and ileal cannulas were assigned to 4 calcium ammonium nitrate addition levels: control, low, medium, and high [0, 5.3, 13.6, and 21.1g of nitrate/kg of dry matter (DM), respectively]. Diets were made isonitrogenous by replacing urea. Cows were fed ad libitum and, after a 6-d period of gradual introduction of nitrate, adapted to the corn-silage-based total mixed ration (forage:concentrate ratio 50:50 on DM basis) for 16d before sampling. Digesta content from duodenum, ileum, and feces, and rumen liquid were collected, after which methane production and hydrogen emissions were measured in respiration chambers. Methane production [L/kg of dry matter intake (DMI)] linearly decreased with increasing nitrate concentrations compared with the control, corresponding to a reduction of 6, 13, and 23% for the low, medium, and high diets, respectively. Methane production was lowered with apparent efficiencies (measured methane reduction relative to potential methane reduction) of 82.3, 71.9, and 79.4% for the low, medium, and high diets, respectively. Addition of nitrate increased hydrogen emissions (L/kg of DMI) quadratically by a factor of 2.5, 3.4, and 3.0 (as L/kg of DMI) for the low, medium, and high diets, respectively, compared with the control. Blood methemoglobin levels and nitrate concentrations in milk and urine increased with increasing nitrate intake, but did not constitute a threat for animal health and human food safety. Microbial crude protein synthesis and efficiency were unaffected. Total volatile fatty acid concentration and molar proportions of acetate, butyrate, and propionate were unaffected, whereas molar proportions of formate increased. Milk yield, milk composition, DMI and digestibility of DM, organic matter, crude protein, and neutral detergent fiber in rumen, small intestine, hindgut, and total tract were unaffected by addition of nitrate. In conclusion, nitrate lowered methane production linearly with minor effects on rumen fermentation and no effects on nutrient digestibility.

Dietary Nitrate for Methane Mitigation Leads to Nitrous Oxide Emissions from Dairy Cows.

Nitrate supplements to cattle diets can reduce enteric CH emissions. However, if NO metabolism stimulates NO emissions, the effectiveness of dietary NO for CH mitigation will be reduced. We quantified NO emissions as part of a dairy cow feeding experiment in which urea was substituted in nearly iso-N diets with 0, 5, 14 or 21 g NO kg dry matter (DM). The feeding experiment was a Latin square with repetition of Period 1. Each period lasted 4 wk, with CH emission measurements in Week 4 using respiration chambers. During Period 3, NO concentrations in chamber outlet air were monitored semicontinuously during 48 h. High, but fluctuating, NO concentrations were seen at the two highest NO levels (up to between 2 and 5 µL L), and dynamics were linked with recent feed intake. In Periods 4 and 5, NO concentrations and feed intake were determined from all four respiration chambers during two 7-h periods. Emissions of NO coincided with feed intake, again with NO concentrations in the microliter per liter range at the two highest NO intake levels. Neither feed nor excretion of NO via urine were significant sources of NO, indicating that emissions came from the animals. Leakages due to rumen fistulation could also not account for NO emissions. The possibility that NO is produced in the oral cavity is discussed. Nitrous oxide emission factors ranged between 0.7 and 1.0% except in one case at 21 g NO kg DM, where it was 3.4%. When accounting for NO emissions at the highest NO intake level, the overall GHG mitigation effect in two different animal-diet combinations changed from -47 to -40%, and from -19 to -17%, respectively, due to NO emissions.

Methane production and digestion of different physical forms of rapeseed as fat supplements in dairy cows.

The purpose of this experiment was to study the effect of the physical form of rapeseed fat on methane (CH4) mitigation properties, feed digestion, and rumen fermentation. Four lactating ruminal-, duodenal-, and ileal-cannulated Danish Holstein dairy cows (143 d in milk, milk yield of 34.3 kg) were submitted to a 4×4 Latin square design with 4 rations: 1 control with rapeseed meal (low-fat, CON) and 3 fat-supplemented rations with either rapeseed cake (RSC), whole cracked rapeseed (WCR), or rapeseed oil (RSO). Dietary fat concentrations were 3.5 in CON, 5.5 in RSC, 6.2 in WCR, and 6.5% in RSO. The amount of fat-free rapeseed was kept constant for all rations. The forage consisted of corn silage and grass silage and the forage to concentrate ratio was 50:50 on a dry matter basis. Diurnal samples of duodenal and ileal digesta and feces were compiled. The methane production was measured for 4 d in open-circuit respiration chambers. Additional fat reduced the CH4 production per kilogram of dry matter intake and as a proportion of the gross energy intake by 11 and 14%, respectively. Neither the total tract nor the rumen digestibility of organic matter (OM) or neutral detergent fiber were significantly affected by the treatment. Relating the CH4 production to the total-tract digested OM showed a tendency to decrease CH4 per kilogram of digested OM for fat-supplemented rations versus CON. The acetate to propionate ratio was not affected for RSC and WCR but was increased for RSO compared with CON. The rumen ammonia concentration was not affected by the ration. The milk and energy-corrected milk yields were unaffected by the fat supplementation. In conclusion, rapeseed is an appropriate fat source to reduce the enteric CH4 production without affecting neutral detergent fiber digestion or milk production. The physical form of fat did not influence the CH4-reducing effect of rapeseed fat. However, differences in the volatile fatty acid pattern indicate that different mechanisms may be involved.

Technical note: test of a low-cost and animal-friendly system for measuring methane emissions from dairy cows.

Methane is a greenhouse gas with a significant anthropogenic contribution from cattle production. A demand exists for techniques that facilitate evaluation of mitigation strategies for dairy cows. Therefore, a low-cost system facilitating the highest possible animal welfare was constructed and validated. The system uses the same principles as systems for open-circuit indirect calorimetry, but to lower the costs, the chamber construction and air-conditioning system were simpler than described for other open-circuit systems. To secure the highest possible animal welfare, the system is located in the cow's daily environment. The system consists of 4 transparent polycarbonate chambers placed in a square so that the cows are facing each other. The chamber dimensions are 183 (width), 382 (length), and 245 cm (height) with a volume of 17 m(3). Flow and concentrations of O(2), CO(2), CH(4), and H(2) are measured continuously in the outlet. Flow is measured with a mass flow meter, O(2) with a paramagnetic sensor, CO(2) and CH(4) with infrared sensors, and H(2) with an electrochemical sensor. Chamber inlet is placed in the barn and background concentrations may differ between chambers, but delta values between background and outlet concentrations for all chambers were within instrument tolerance. Average recovery rates of CO(2) and CH(4) were (mean ± SD) 101 ± 4 and 99 ± 7%, respectively. This is within the expected tolerance of the whole system (gas sensors and flow meters). Feed dry matter intakes were not affected by confining the animals in chambers, as dry matter intake before and during chamber stay were similar. It was concluded that the system delivers reliable values, and the transparent construction in combination with the location in the barn environment prevent negative impact on animal welfare and, thereby, data quality.

Methane production and diurnal variation measured in dairy cows and predicted from fermentation pattern and nutrient or carbon flow.

Many feeding trials have been conducted to quantify enteric methane (CH(4)) production in ruminants. Although a relationship between diet composition, rumen fermentation and CH(4) production is generally accepted, the efforts to quantify this relationship within the same experiment remain scarce. In the present study, a data set was compiled from the results of three intensive respiration chamber trials with lactating rumen and intestinal fistulated Holstein cows, including measurements of rumen and intestinal digestion, rumen fermentation parameters and CH(4) production. Two approaches were used to calculate CH(4) from observations: (1) a rumen organic matter (OM) balance was derived from OM intake and duodenal organic matter flow (DOM) distinguishing various nutrients and (2) a rumen carbon balance was derived from carbon intake and duodenal carbon flow (DCARB). Duodenal flow was corrected for endogenous matter, and contribution of fermentation in the large intestine was accounted for. Hydrogen (H(2)) arising from fermentation was calculated using the fermentation pattern measured in rumen fluid. CH(4) was calculated from H(2) production corrected for H(2) use with biohydrogenation of fatty acids. The DOM model overestimated CH(4)/kg dry matter intake (DMI) by 6.1% (R(2)=0.36) and the DCARB model underestimated CH(4)/kg DMI by 0.4% (R(2)=0.43). A stepwise regression of the difference between measured and calculated daily CH(4) production was conducted to examine explanations for the deviance. Dietary carbohydrate composition and rumen carbohydrate digestion were the main sources of inaccuracies for both models. Furthermore, differences were related to rumen ammonia concentration with the DOM model and to rumen pH and dietary fat with the DCARB model. Adding these parameters to the models and performing a multiple regression against observed daily CH(4) production resulted in R 2 of 0.66 and 0.72 for DOM and DCARB models, respectively. The diurnal pattern of CH(4) production followed that of rumen volatile fatty acid (VFA) concentration and the CH(4) to CO(2) production ratio, but was inverse to rumen pH and the rumen hydrogen balance calculated from 4×(acetate+butyrate)/2×(propionate+valerate). In conclusion, the amount of feed fermented was the most important factor determining variations in CH(4) production between animals, diets and during the day. Interactions between feed components, VFA absorption rates and variation between animals seemed to be factors that were complicating the accurate prediction of CH(4). Using a ruminal carbon balance appeared to predict CH(4) production just as well as calculations based on rumen digestion of individual nutrients.

Milk production is unaffected by replacing barley or sodium hydroxide wheat with maize cob silage in rations for dairy cows.

Starch is an important energy-providing nutrient for dairy cows that is most commonly provided from cereal grains. However, ruminal fermentation of large amounts of easily degradable starch leads to excessive production and accumulation of volatile fatty acids (VFA). VFA not only play a vital role in the energy metabolism of dairy cows but are also the main cause of ruminal acidosis and depressed feed intake. The aim of the present study was to compare maize cob silage (MCS) as an energy supplement in rations for dairy cows with highly rumen-digestible rolled barley and with sodium hydroxide wheat (SHW), which has a higher proportion of by-pass starch than barley. Two studies were carried out: (1) a production study on 45 Danish Holstein cows and (2) an intensive study to determine digestibilities, rumen fermentation patterns and methane emission using three rumen-cannulated Danish Holstein cows. Both studies were organised as a 3×3 Latin square with three experimental periods and three different mixed rations. The rations consisted of grass-clover silage and maize silage (~60% of dry matter (DM)), rapeseed cake, soybean meal, sugar beet pulp and one of three different cereals as a major energy supplement: MCS, SHW or rolled barley (~25% of DM). When MCS replaced barley or SHW as an energy supplement in the mixed rations, it resulted in a lower dry matter intake; however, the apparent total tract digestibilities of DM, organic matter, NDF, starch and protein were not different between treatments. The energy-corrected milk yield was unaffected by treatment. The fat content of the milk on the MCS ration was not different from the SHW ration, whereas it was higher on the barley ration. The protein content of the milk decreased when MCS was used in the ration compared with barley and SHW. From ruminal VFA patterns and pH measures, it appeared that MCS possessed roughage qualities with respect to rumen environment, while at the same time being sufficiently energy rich to replace barley and SHW as a major energy supplement for milk production. The environmental impact, expressed as methane emissions, was not different when comparing MCS, SHW and barley.