Selection for more feed efficient dairy cows is key to improving sustainability and profitability of dairy production; however, underlying mechanisms contributing to individual animal feed efficiency are not fully understood. The objective of this study was to identify circulating metabolites, and pathways associated with those metabolites, that differ between efficient and inefficient Holstein dairy cows using targeted metabolite quantification and untargeted metabolomics. The top and bottom fifteen percent of cows (n = 28/group) with the lowest and highest residual feed intake in mid-lactation feed efficiency trials were grouped retrospectively as high-efficient (HE) and low-efficient (LE). Blood samples were collected for quantification of energy metabolites, markers of hepatic function, and acylcarnitines, in addition to a broader investigation using untargeted metabolomics. Short-chain acylcarnitines, C3-acylcarnitine, and C4-acylcarntine were lower in HE cows (n = 18/group). Untargeted metabolomics and multivariate analysis identified thirty-nine differential metabolites between HE and LE (n = 8/group), of which twenty-five were lower and fourteen were higher in HE. Pathway enrichment analysis indicated differences in tryptophan metabolism. Combined results from targeted metabolite quantification and untargeted metabolomics indicate differences in fatty acid and amino acid metabolism between HE and LE cows. These differences may indicate post-absorptive nutrient use efficiency as a contributor to individual animal variation in feed efficiency.

Peripartum rumen-protected choline (RPC) supplementation is beneficial for cow health and production, yet the optimal dose is unknown. In vivo and in vitro supplementation of choline modulates hepatic lipid, glucose, and methyl donor metabolism. The objective of this experiment was to determine the effects of increasing the dose of prepartum RPC supplementation on milk production and blood biomarkers. Pregnant multiparous Holstein cows (n = 116) were randomly assigned to one of 4 prepartum choline treatments that were fed from -21 d relative to calving (DRTC) until calving. From calving until +21 DRTC, cows were fed diets targeting 0 g/d choline ion (control, CTL) or the recommended dose (15 g/d choline ion; RD) of the same RPC product that they were fed prepartum. The resulting treatments targeted: (1) 0 g/d pre- and postpartum [0.0 ± 0.000 choline ion, percent of dry matter (%DM); CTL]; (2) 15 g/d pre- and postpartum of choline ion from an established product (prepartum: 0.10 ± 0.004 choline ion, %DM; postpartum: 0.05 ± 0.004 choline ion, %DM; ReaShure, Balchem Corp.; RPC1_RD▸RD); (3) 15 g/d pre- and postpartum of choline ion from a concentrated RPC prototype (prepartum: 0.09 ± 0.004 choline ion, %DM; postpartum: 0.05 ± 0.003 choline ion, %DM; RPC2, Balchem Corp.; RPC2_RD▸RD); or (4) 22 g/d prepartum and 15 g/d postpartum from RPC2 [prepartum: 0.13 ± 0.005 choline ion, %DM; postpartum: 0.05 ± 0.003 choline ion, %DM; high prepartum dose (HD), RPC2_HD▸RD]. Treatments were mixed into a total mixed ration, and cows had ad libitum access via a roughage intake control system (Hokofarm Group). From calving to +21 DRTC, all cows were fed a common base diet and treatments were mixed into the total mixed ration (supplementation period, SP). Thereafter, all cows were fed a common diet (0 g/d choline ion) until +100 DRTC (postsupplementation period, postSP). Milk yield was recorded daily and composition analyzed weekly. Blood samples were obtained via tail vessel upon enrollment, approximately every other day from -7 to +21 DRTC, and at +56 and +100 DRTC. Feeding any RPC treatment reduced prepartum dry matter intake compared with CTL. During the SP, no evidence for a treatment effect on energy-corrected milk (ECM) yield was found, but during the postSP, RPC1_RD▸RD and RPC2_RD▸RD treatments tended to increase ECM, protein, and fat yields. During the postSP, the RPC1_RD▸RD and RPC2_RD▸RD treatments tended to increase, and RPC2_HD▸RD increased, the de novo proportion of total milk fatty acids. During the early lactation SP, RPC2_HD▸RD tended to increase plasma fatty acids and β-hydroxybutyrate concentrations, and RPC1_RD▸RD and RPC2_RD▸RD reduced blood urea nitrogen concentrations compared with CTL. The RPC2_HD▸RD treatment reduced early lactation serum lipopolysaccharide binding protein compared with CTL. Overall, peripartum RPC supplementation at the recommended dose tended to increase ECM yield postSP, but no evidence was seen of an additional benefit on milk production with an increased prepartum dose of choline ion. The effects of RPC on metabolic and inflammatory biomarkers support the potential for RPC supplementation to affect transition cow metabolism and health and may support the production gains observed.

Feeding pregnant cows rumen-protected choline (RPC) may have the potential to affect the growth and health of offspring, but little is known about the optimal dose, or the potential mechanisms of action. The objectives of this experiment were to 1) determine if increasing RPC supplementation during late gestation in multiparous Holstein cows would improve calf growth and 2) determine if maternal choline supplementation alters global DNA methylation patterns. Pregnant multiparous Holstein cows (n = 116) were randomly assigned to diets targeting 0g choline ion (0.0 ± 0.000 choline ion, %DM, control; CTL), 15g of choline ion (recommended dose; RD) from an established RPC product (0.10 ± 0.004 choline ion, %DM, RPC1_RD; ReaShure, Balchem Corp.; positive control), or 15g (0.09 ± 0.004 choline ion, %DM, RPC2_RD) or 22g (0.13 ± 0.005 choline ion, %DM, high dose; RPC2_HD) of choline ion from a concentrated RPC prototype (RPC2; Balchem Corp.). Treatments were mixed into a total mixed ration and cows had ad libitum access via a roughage intake control system (Hokofarm Group, Marknesse, Netherlands). All female Holstein (n = 49) and Holstein × Angus calves (male, n = 18; female, n = 30) were enrolled and fed colostrum from a cow within the same treatment. Holstein calves and Holstein × Angus calves were fed an accelerated and traditional milk replacer program, respectively, and offered ad libitum access to calf starter. Jugular vein blood samples were collected, and body weight was measured at 7, 14, 28, 42, and 56 d of age. Categorical treatment and continuous effects of actual prepartum maternal choline ion intake were analyzed using mixed effect models. An interaction of treatment with sex, nested within breed, resulted in any choline treatment increasing the proportion of methylated whole blood DNA in male, but not female calves. Although 37% of Holstein calves across all treatments experienced abomasal bloat, no evidence for differences in health measurements (signs of respiratory disease and fecal consistency) were observed across treatments. During the first 2 wk of life in Holstein calves, RPC2_HD tended to increase average daily gain (ADG) and feed efficiency (FE) compared with CTL and increasing maternal choline ion intake linearly increased ADG and FE. Maternal choline supplementation increased plasma glucose compared with CTL, while increasing serum insulin-like growth factor-1 and decreasing serum lipopolysaccharide binding protein at 7 d of age in Holstein calves. In Holstein × Angus calves, the effect of treatment on ADG tended to interact with sex: in males, RPC2_HD increased ADG after 2 wk of life compared with CTL, without evidence of a treatment effect in female calves. Increasing maternal choline ion intake linearly increased ADG after 2 wk of age in male Holstein × Angus calves, while quadratically increasing FE in both sexes. Altered global DNA methylation patterns in male Holstein × Angus calves, and changes in blood metabolites in Holstein calves, provide 2 potential mechanisms for observed improvements in calf growth. Continuous treatment models demonstrated that the effects of maternal choline supplementation are sensitive to the amount of maternal choline ion intake, with greater benefit to calves observed at higher maternal intakes.

Quantifying dry matter intake (DMI) in lactating dairy cows is important for determining feed efficiency; however, there are no methods for economically quantifying individual cow DMI on dairy farms where cows are group-fed. Attempts have been made to model DMI using cow factors, milk production, milk infrared spectra, and behavioral sensors with reasonable success. Other data streams are available on the farm that may contribute to DMI predictions. In this study, our objective was to model DMI with multiple linear regression using data from a single point-in-time that can easily be accessed on-farm. Candidate predictor variables included cow descriptors, milk yield and composition, milk fatty acid profile, and production and efficiency predicting transmitting abilities (PTA). Observations of DMI were obtained from 350 cows across 6 cohorts using individual feed bunks. The cow to bunk ratio was 2:1, with an overall bunk occupation rate of 32% throughout the day. The following models were developed sequentially with milk data obtained from a single morning milking and other data from the same day: model B (production, metabolic body weight, body condition score, lactation category, and week of lactation), model BC [model B + fatty acid (FA) content], model BY (model B + FA yield), model BPE (model B + production and efficiency PTA), model BYP (model BY + production PTA), model BYE (model BY + efficiency PTA), and model BYPE (model BY + production and efficiency PTA). Outcome variables predicted in these models were the DMI on the previous day or current day relative to the morning milk sample. The predictions for DMI on the previous day outperformed current day DMI in every model for which they were both determined. Addition of milk FA and PTA as candidate predictor variable types to the models resulted in enhanced predictive ability, with incremental enhancements when combined. The most robust model (BYPE) included cow descriptors, protein and FA yields, and PTA for milk and residual feed intake. Model BYPE described 21 to 32% more of the variation in DMI (based on concordance correlation coefficient) than when other common DMI models were applied to the same data set. Overall, reasonable performance of models including single point-in-time cow descriptors, milk and FA production, and production and efficiency PTA commonly available to dairy farmers through dairy herd improvement programs offer an opportunity for on-farm prediction of DMI, yet further improvement may be possible.

The objective of this study was to investigate the relationships between postpartum health disorders and mid-lactation performance, feed efficiency, and sensor-derived behavioral traits. Multiparous cows (n = 179) were monitored for health disorders for 21 days postpartum and enrolled in a 45-day trial between 50 to 200 days in milk, wherein feed intake, milk yield and components, body weight, body condition score, and activity, lying, and feeding behaviors were recorded. Feed efficiency was measured as residual feed intake and the ratio of fat- or energy-corrected milk to dry matter intake. Cows were classified as either having hyperketonemia (HYK; n = 72) or not (n = 107) and grouped by frequency of postpartum health disorders: none (HLT; n = 94), one (DIS; n = 63), or ≥2 (DIS+; n = 22). Cows that were diagnosed with HYK had higher mid-lactation yields of fat- and energy-corrected milk. No differences in feed efficiency were detected between HYK or health status groups. Highly active mid-lactation time was higher in healthy animals, and rumination time was lower in ≥4th lactation cows compared with HYK or DIS and DIS+ cows. Differences in mid-lactation behaviors between HYK and health status groups may reflect the long-term impacts of health disorders. The lack of a relationship between postpartum health and mid-lactation feed efficiency indicates that health disorders do not have long-lasting impacts on feed efficiency.

Improving dairy cow feed efficiency is critical to the sustainability and profitability of dairy production, yet the underlying mechanisms that contribute to individual cow variation in feed efficiency are not fully understood. The objectives of this study were to (1) identify genes and associated pathways that are altered in cows with high- or low-residual feed intake (RFI) using RNA sequencing, and (2) determine if rumen-protected choline supplementation during mid-lactation would influence performance or feed efficiency. Mid-lactation (134 ± 20 days in milk) multiparous Holstein cows were randomly assigned to either supplementation of 0 g/d supplementation (CTL; n = 32) or 30 g/d of a rumen-protected choline product (RPC; 13.2 g choline ion; n = 32; Balchem Corp., New Hampton, NY, USA). Residual feed intake was determined as dry matter intake regressed on milk energy output, days in milk, body weight change, metabolic body weight, and dietary treatment. The 12 cows with the highest RFI (low feed efficient; LE) and 12 cows with the lowest RFI (high feed efficient; HE), balanced by dietary treatment, were selected for blood, liver, and muscle analysis. No differences in production or feed efficiency were detected with RPC supplementation, although albumin was greater and arachidonic acid tended to be greater in RPC cows. Concentrations of β-hydroxybutyrate were greater in HE cows. Between HE and LE, 268 and 315 differentially expressed genes in liver and muscle tissue, respectively, were identified through RNA sequencing. Pathway analysis indicated differences in cell cycling, oxidative stress, and immunity in liver and differences in glucose and fatty acid pathways in muscle. The current work indicates that unique differences in liver and muscle post-absorptive nutrient metabolism contribute to sources of variation in feed efficiency and that differences in amino acid and fatty acid oxidation, cell cycling, and immune function should be further examined.