Effects of feeding whole-cracked rapeseeds, nitrate, and 3-nitrooxypropanol on composition and functional properties of the milk fat fraction from Danish Holstein cows.

The aim of this study was to determine the individual and combined effects of supplementing fat (FAT), nitrate (NITRATE) and 3-nitrooxypropanol (3-NOP) on compositional and functional properties of milk fat. An 8 × 8 incomplete Latin square design was conducted with 48 lactating Danish Holstein cows over 6 periods of 21 d each. Eight diets were 2 × 2 × 2 factorially arranged: FAT (30 or 63 g crude fat/kg DM), NITRATE (0 or 10 g nitrate/kg DM), and 3-NOP (0 or 80 mg 3-NOP/kg DM) and cows were fed ad libitum. Milk samples were analyzed for general composition, fatty acids (FA) and thermal properties of milk fat. Milk fat content was decreased by FAT and NITRATE and increased by 3-NOP. The changes in FA composition were mainly driven by the FAT × 3-NOP interaction. FAT shifted milk FA composition toward lower content of saturated FA (SFA) and greater contents of mono- and poly-unsaturated FA (MUFA and PUFA), whereas these effects of FAT were smaller in combination with 3-NOP. However, 3-NOP had no effects on SFA, MUFA and PUFA in low fat diets. FAT lowered solid fat content in milk fat because of decreased SFA content. The onset crystallization temperature of milk fat was decreased by 3-NOP when supplemented in low fat diets. According to the FAT × 3-NOP interaction, supplementation of fat without 3-NOP shifted peak temperature of low melting fraction of milk fat toward low temperature as a result of decreased proportion of C16:0, and increased proportions of C18:1 cis-9, C18:1 trans-11, C18:2 cis-9, and CLA cis-9,trans-11. In conclusion, no additive effects were observed among FAT, NITRATE and 3-NOP on chemical and thermal properties of milk fat and fat supplementation largely changed milk FA composition and in turn affected the thermal properties of milk fat.

Effects of feeding whole-cracked rapeseeds, nitrate, and 3-nitrooxypropanol on protein composition, minerals, and vitamin B in milk from Danish Holstein cows.

The present study was conducted to assess the individual or combined effects of feeding dietary fat (whole-cracked rapeseed), nitrate, and 3-nitrooxypropanol (3-NOP) on protein profile, mineral composition, B vitamins, and nitrate residues in milk from dairy cows. Forty-eight Danish Holstein cows used in an 8 × 8 incomplete Latin square design were fed 8 factorially arranged diets ((30 or 63 g crude fat/kg DM) × (0 or 10 g nitrate/kg DM) × (0 or 80 mg 3-NOP/kg DM)) over 6 periods of 21 d each. In each period, milk samples were collected from individual cows during the third week by pooling milk obtained from 4 consecutive milkings, and analyzed for protein profile including protein modifications, mineral composition, riboflavin, cobalamin, and presence of nitrate residues. Fat supplementation led to an increase in the phosphorylation degree of αS1-CN by 8.5% due to a decreased relative proportion of αS1-CN 8P and an increased relative proportion of αS1-CN 9P and further to a decrease in the relative proportion of αS2-CN by 2.4%. Additionally, fat supplementation decreased the relative proportions of glycosylated and unglycosylated forms of κ-CN, consequently leading to a 3.6% decrease in total κ-CN. In skim milk, K, Ca, P, and Mg concentrations were altered by individual use of fat, nitrate, and 3-NOP. Feeding nitrate resulted in a 5.4% increase in riboflavin concentration in milk while supplementing 3-NOP increased cobalamin concentration in milk by 21.1%. The nitrate concentration in milk was increased upon feeding nitrate however, this increased concentration was well below the maximum permissible limit of nitrate in milk (<50 mg/L). In conclusion, no major changes were observed in milk protein, and mineral compositions by feeding fat, nitrate, and 3-NOP to dairy cows while the increased riboflavin and cobalamin by nitrate and 3-NOP, respectively, could be of beneficial nutritional value for milk consumers.

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 develop different models to be used for different scenarios, depending on available data. An international data set 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 data set. 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 data set: 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 data sets either with 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 over-prediction and under-prediction of CO2 production when the models were evaluated on the GF and the RC test data set, respectively. To address this bias, the 3 models were evaluated on a modified test data set, 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 data sets. By 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.

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.

Nitrogen isotopic discrimination as a biomarker of between-cow variation in the efficiency of nitrogen utilization for milk production: A meta-analysis.

Estimating the efficiency of N utilization for milk production (MNE) of individual cows at a large scale is difficult, particularly because of the cost of measuring feed intake. Nitrogen isotopic discrimination (Δ15N) between the animal (milk, plasma, or tissues) and its diet has been proposed as a biomarker of the efficiency of N utilization in a range of production systems and ruminant species. The aim of this study was to assess the ability of Δ15N to predict the between-animal variability in MNE in dairy cows using an extensive database. For this, 20 independent experiments conducted as either changeover (n = 14) or continuous (n = 6) trials were available and comprised an initial data set of 1,300 observations. Between-animal variability was defined as the variation observed among cows sharing the same contemporary group (CG; individuals from the same experimental site, sampling period, and dietary treatment). Milk N efficiency was calculated as the ratio between mean milk N (grams of N in milk per day) and mean N intake (grams of N intake per day) obtained from each sampling period, which lasted 9.0 ± 9.9 d (mean ± SD). Samples of milk (n = 604) or plasma (n = 696) and feeds (74 dietary treatments) were analyzed for natural 15N abundance (δ15N), and then the N isotopic discrimination between the animal and the dietary treatment was calculated (Δ15n = δ15Nanimal - δ15Ndiet). Data were analyzed through mixed-effect regression models considering the experiment, sampling period, and dietary treatment as random effects. In addition, repeatability estimates were calculated for each experiment to test the hypothesis of improved predictions when MNE and Δ15N measurements errors were lower. The considerable protein mobilization in early lactation artificially increased both MNE and Δ15N, leading to a positive rather than negative relationship, and this limited the implementation of this biomarker in early lactating cows. When the experimental errors of Δ15N and MNE decreased in a particular experiment (i.e., higher repeatability values), we observed a greater ability of Δ15N to predict MNE at the individual level. The predominant negative and significant correlation between Δ15N and MNE in mid- and late lactation demonstrated that on average Δ15N reflects MNE variations both across dietary treatments and between animals. The root mean squared prediction error as a percentage of average observed value was 6.8%, indicating that the model only allowed differentiation between 2 cows in terms of MNE within a CG if they differed by at least 0.112 g/g of MNE (95% confidence level), and this could represent a limitation in predicting MNE at the individual level. However, the one-way ANOVA performed to test the ability of Δ15N to differentiate within-CG the top 25% from the lowest 25% individuals in terms of MNE was significant, indicating that it is possible to distinguish extreme animals in terms of MNE from their N isotopic signature, which could be useful to group animals for precision feeding.

Prediction of nitrogen excretion from data on dairy cows fed a wide range of diets compiled in an intercontinental database: A meta-analysis.

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.

Effect of protein concentrate mixtures and dietary addition of exogenous phytase on major milk minerals and proteins, including casein phosphorylation.

Variations in major milk minerals, proteins, and their posttranslational modifications are largely under genetic influence, whereas the effect of nongenetic factors is less studied. Through a controlled feeding experiment (incomplete balanced Latin square design), the effect of concentrate mixtures, based on fava beans, rapeseed meal, or soybean meal as main P and protein sources, on milk composition was examined under typical Danish management conditions. Concentrations of P, Ca, and Mg, together with proteomics for relative quantification of major milk proteins and their isoforms, were analyzed in milk samples from 24 cows sampled in 4 periods. Each cow was fed 1 of the 3 diets in each period with or without addition of exogenous phytase. Cows were blocked by lactation stage into early and mid-lactation (23.3 ± 6.7 and 176 ± 15 d in milk, respectively, at the beginning of the experiment, mean ± standard deviation). Significant effects of feed concentrate mixture were observed for milk protein concentration, milk urea nitrogen, citrate, and the percentage of mixed and preformed fatty acids as well as mineral composition, and their distributions within micellar or serum phases. Furthermore, relative contents of αS1-casein (CN) 9P form and unglycosylated κ-CN and thereby phosphorylation degree of αS1-CN (PD) and the glycosylation degree of κ-CN were found to be significantly affected by these diets. To our knowledge, we are the first to document that feed concentrate mixture can affect the relative concentrations of αS1-CN phosphorylation isoforms in milk, and the results suggested an effect on αS1-CN 9P and PD, but not on αS1-CN 8P. Furthermore, although only significant for αS1-CN 8P, we found a lower relative concentration of αS1-CN 8P and higher αS1-CN 9P (and thus higher PD) in milk from cows in mid compared with early lactation. Also, protein concentration and concentration of Mg in skim milk and serum as well as relative concentration of α-lactalbumin were found to be significantly affected by lactation stage. Addition of dietary exogenous phytase only had a minor effect on milk composition or functionality with significant effect detected for α-lactalbumin and micellar Mg concentration.

Fava beans can substitute soybean meal and rapeseed meal as protein source in diets for lactating dairy cows.

The effect of replacing mixtures of wheat and soybean meal and wheat and rapeseed meal by toasted fava beans, and the effect of toasting fava beans on feed intake, milk yield, and composition of milk and feces were investigated using 40 Holstein cows in each of two 4 × 4 Latin square design trials conducted simultaneously. In trial 1, the 4 treatment concentrates were untreated fava beans, toasted fava beans, 42% soybean meal + 58% rolled wheat, and a 21, 29, and 50% mix of soybean meal, rolled wheat, and toasted fava beans, respectively [on dry matter (DM) basis]. In trial 2, the 4 experimental treatments were untreated fava beans, toasted fava beans, 64% rapeseed meal + 36% rolled wheat, and a 32, 18, and 50% mix of rapeseed meal, rolled wheat, and toasted fava beans, respectively (on DM basis). In each trial, 16 primiparous and 24 multiparous cows were fed the treatment concentrates as part of a partial mixed ration, of which the forage consisted of 50% corn silage and 50% grass-clover silage. Substitution of soybean meal and wheat or rapeseed meal and wheat with toasted fava beans did not affect total DM intake, and no linear effects were observed on milk yield or energy-corrected milk (ECM) yield. However, in trial 2, a quadratic effect was observed on milk yield when substituting rapeseed meal and wheat with toasted fava beans. In both trials, substitution of soybean meal and wheat or rapeseed meal and wheat with toasted fava beans increased milk lactose concentration and decreased milk protein yield and concentration of protein in milk. In both trials, fecal concentration of starch increased linearly when substituting soybean meal and wheat or rapeseed meal and wheat with toasted fava beans. In trial 2, fecal concentration of P decreased when substituting rapeseed meal and wheat with toasted fava beans. In situ investigations showed increased rumen undegradable protein concentration and thereby increased estimated metabolizable protein supply when toasting fava beans. However, in both trials, milk protein yield and concentration decreased when cows were fed toasted compared with untreated fava beans. Furthermore, when cows were fed toasted compared with untreated fava beans in trial 1, milk yield, ECM yield, and nitrogen efficiency decreased. We conclude that toasted fava beans could substitute soybean meal and wheat or rapeseed meal and wheat with regard to ECM yield. However, milk protein yield decreased when substituting soybean meal and wheat or rapeseed meal and wheat with toasted fava beans. Compared with untreated fava beans, toasting had no positive effect on milk production and nitrogen efficiency.

The effects of energy metabolism variables on feed efficiency in respiration chamber studies with lactating dairy cows.

The objective of the present study was to investigate factors related to variation in feed efficiency (FE) among cows. Data included 841 cow/period observations from 31 energy metabolism studies assembled across 3 research stations. The cows were categorized into low-, medium-, and high-FE groups according to residual feed intake (RFI), residual energy-corrected milk (RECM), and feed conversion efficiency (FCE). Mixed model regression was conducted to identify differences among the efficiency groups in animal and energy metabolism traits. Partial regression coefficients of both RFI and RECM agreed with published energy requirements more closely than cofficients derived from production experiments. Within RFI groups, efficient (Low-RFI) cows ate less, had a higher digestibility, produced less methane (CH4) and heat, and had a higher efficiency of metabolizable energy (ME) utilization for milk production. High-RECM (most efficient) cows produced 6.0 kg/d more of energy-corrected milk (ECM) than their Low-RECM (least efficient) contemporaries at the same feed intake. They had a higher digestibility, produced less CH4 and heat, and had a higher efficiency of ME utilization for milk production. The contributions of improved digestibility, reduced CH4, and reduced urinary energy losses to increased ME intake at the same feed intake were 84, 12, and 4%, respectively. For both RFI and RECM analysis, increased metabolizability contributed to approximately 35% improved FE, with the remaining 65% attributed to the greater efficiency of utilization of ME. The analysis within RECM groups suggested that the difference in ME utilization was mainly due to the higher maintenance requirement of Low-RECM cows compared with Medium- and High-RECM cows, whereas the difference between Medium- and High-RECM cows resulted mainly from the higher efficiency of ME utilization for milk production in High-RECM cows. The main difference within FCE (ECM/DMI) categories was a greater (8.2 kg/d) ECM yield at the expense of mobilization in High-FCE cows compared with Low-FCE cows. Methane intensity (CH4/ECM) was lower for efficient cows than for inefficient cows. The results indicated that RFI and RECM are different traits. We concluded that there is considerable variation in FE among cows that is not related to dilution of maintenance requirement or nutrient partitioning. Improving FE is a sustainable approach to reduce CH4 production per unit of product, and at the same time improve the economics of milk production.

Phenotypic modeling of residual feed intake using physical activity and methane production as energy sinks.

Residual feed intake (RFI) is a measure of feed efficiency in dairy cattle. This study modeled phenotypic RFI of first- and second-parity Holstein and Jersey dairy cows within 9 lactation segments (consecutive segments of 4 wk each) covering the first 36 lactation weeks. We aimed to evaluate physical activity and daily methane production as additional energy sinks in the estimation of RFI, to examine the correlations of RFI among the first 36 wk of lactation (WOL), and to evaluate whether parities and breeds show similar results. Records for first-parity Holstein (n = 449), second-parity Holstein (n = 298), first-parity Jersey (n = 195), and second-parity Jersey cows (n = 146) were used. Model 1 included the following energy sinks: energy-corrected milk yield, metabolic body weight (BW), body condition score (BCS), daily changes in BW (ΔBW) and BCS (ΔBCS), and physical activity. Model 2 was based on a subset of the data and only for Holstein cows, and included the same energy sinks as Model 1, plus daily methane production. The trajectories of segment-specific partial regression coefficients (PRC) of DMI on activity were similar across parities but differed slightly between breeds. For daily methane production, the trajectory in PRC decreased over lactation segments for first- and second-parity Holstein cows. The trajectories in PRC of DMI on energy-corrected milk yield, metabolic BW, BCS, and ΔBW were generally similar across parities, except for ΔBCS. Activity accounted for on average 7.3, 6.8, 7.2, and 6.4% of DMI for first-parity Holsteins, second-parity Holsteins, first-parity Jerseys, and second-parity Jerseys, respectively. Methane losses accounted for 8.7% and 8.5% of DMI for first- and second-parity Holstein cows, respectively. Repeatability estimates for RFI over 36 WOL for Model 1 were 0.63 for first-parity Holsteins, 0.65 for second-parity Holsteins, 0.76 for first-parity Jerseys, and 0.80 for second-parity Jerseys. For Model 2, the estimates were 0.59 and 0.61 for first- and second-parity Holstein cows, respectively. Correlations of RFI between WOL varied in strength, with weak correlations for the first 2 to 3 WOL with other WOL. In conclusion, physical activity and daily methane production accounted for part of DMI, and RFI of dairy cattle is not the same trait throughout lactation.

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.

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.

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.

Examining trends in the diversity of the U.S. National Institutes of Health participating and funded workforce.

Here, we use recent U.S. National Institutes of Health (NIH) data to document trends in the NIH-funded workforce over time. Consistent with previous studies that were initiated by NIH, we find that the number of scientists funded on competing R01-equivalent (R01 Eq.) and research project grants (RPGs) increased 2-5% per year between 2009 and 2016. Primary beneficiaries of this growth were experienced investigators (Exp), whereas the share of funding awarded to early-stage investigators (ESIs) and new investigators (NIs) declined. The decline occurred even after NIH instituted the New and Early-Stage Investigator policy in 2009. When we evaluate the investigator pool, we find that women and racial and ethnic minorities represent a higher percentage of NIs and ESIs relative to Exp. Thus, trends of diminishing support for NIs and ESIs may negatively impact the diversity of the current and future biomedical research workforce. We find some recent gains among women and Hispanics as part of the applicant and awardee pool for both R01 Eq. and RPGs, but significant, large gaps persist among nationally underrepresented racial minorities. Our findings suggest a need to prioritize investments and support of ESIs and NIs, groups in which women and racial and ethnic minorities represent a larger proportion of the applicant pool, to enhance diversity in the NIH-funded workforce.-Nikaj, S., Roychowdhury, D., Lund, P. K., Matthews, M., Pearson, K. Examining trends in the diversity of the U.S. National Institutes of Health participating and funded workforce.

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.

Individual milk fatty acids are potential predictors of enteric methane emissions from dairy cows fed a wide range of diets: Approach by meta-analysis.

There is a need to quantify methane (CH4) emissions with alternative methods. For the past decade, milk fatty acids (MFA) could be used as proxies to predict CH4 emissions from dairy cows because of potential common rumen biochemical pathways. However, equations have been developed based on a narrow range of diets and with limited data. The objectives of this study were to (1) construct a set of empirical models based on individual data of CH4 emissions and MFA from a large number of lactating dairy cows fed a wide range of diets; (2) further increase the models' level of complexity (from farm to research level) with additional independent variables such as dietary chemical composition (organic matter, neutral detergent fiber, crude protein, starch, and ether extract), dairy performance (milk yield and composition), and animal characteristics (days in milk or body weight); and (3) evaluate the performance of the developed models on independent data sets including measurements from individual animals or average measurements of groups of animals. Prediction equations based only on MFA [C10:0, iso C17:0 + trans-9 C16:1,cis-11 C18:1, and trans-11,cis-15 C18:2 for CH4 production (g/d); iso C16:0, cis-11 C18:1, trans-10 C18:1, and cis-9,cis-12 C18:2 for CH4 yield (g/kg of dry matter intake, DMI); and iso C16:0, cis-15 C18:1, and trans-10 + trans-11 C18:1 for CH4 intensity (g/kg of milk)] had a root mean squared error of 65.1 g/d, 2.8 g/kg of DMI, and 2.9 g/kg of milk, respectively, whereas complex equations that additionally used DMI, dietary neutral detergent fiber, ether extract, days in milk, and body weight had a lower root mean squared error of 46.6 g/d, 2.6 g/kg of DMI, and 2.7 g/kg of milk, respectively). External evaluation with individual or mean data not used for equation development led to variable results. When evaluations were performed using individual cow data from an external data set, accurate predictions of CH4 production (g/d) were obtained using simple equations based on MFA. Better performance was observed on external evaluation with individual data for the simple equation of CH4 production (g/d, based on MFA), whereas better performance was observed on external evaluation mean data for the simple equation of CH4 yield (g/kg of DMI). The performance of evaluation of the models is dependent on the domain of validity of the evaluation data sets used (individual or mean).

Functional Transcriptomics in Diverse Intestinal Epithelial Cell Types Reveals Robust MicroRNA Sensitivity in Intestinal Stem Cells to Microbial Status.

Gut microbiota play an important role in regulating the development of the host immune system, metabolic rate, and at times, disease pathogenesis. The factors and mechanisms that mediate interactions between microbiota and the intestinal epithelium are not fully understood. We provide novel evidence that microbiota may control intestinal epithelial stem cell (IESC) proliferation in part through microRNAs (miRNAs). We demonstrate that miRNA profiles differ dramatically across functionally distinct cell types of the mouse jejunal intestinal epithelium and that miRNAs respond to microbiota in a highly cell type-specific manner. Importantly, we also show that miRNAs in IESCs are more prominently regulated by microbiota compared with miRNAs in any other intestinal epithelial cell subtype. We identify miR-375 as one miRNA that is significantly suppressed by the presence of microbiota in IESCs. Using a novel method to knockdown gene and miRNA expression ex vivo enteroids, we demonstrate that we can knock down gene expression in Lgr5+ IESCs. Furthermore, when we knock down miR-375 in IESCs, we observe significantly increased proliferative capacity. Understanding the mechanisms by which microbiota regulate miRNA expression in IESCs and other intestinal epithelial cell subtypes will elucidate a critical molecular network that controls intestinal homeostasis and, given the heightened interest in miRNA-based therapies, may offer novel therapeutic strategies in the treatment of gastrointestinal diseases associated with altered IESC function.