Identification of genomic regions and genes associated with subclinical ketosis in periparturient dairy cows.

Subclinical ketosis (SCK) is a prevalent metabolic disorder that occurs during the transition to lactation period. It is defined as a high blood concentration of ketone bodies (beta-hydroxybutyric acid f ≥ 1.2 mmol/L) within the first few weeks of lactation, and often presents without clinical signs. SCK is mainly caused by negative energy balance (NEB). The objective of this study is to identify single nucleotide polymorphisms (SNPs) associated with SCK using genome-wide association studies (GWAS), and to predict the biological functions of proximal genes using gene-set enrichment analysis (GSEA). Blood samples were collected from 112 Holstein cows between 5 and 18 days postpartum to determine the incidence of SCK. Genomic DNA extracted from both SCK and healthy cows was examined using the Illumina Bovine SNP50K BeadChip for genotyping. GWAS revealed 194 putative SNPs and 163 genes associated with those SNPs. Additionally, GSEA showed that the genes retrieved by Database for Annotation, Visualization, and Integrated Discovery (DAVID) belonged to calcium signaling, starch and sucrose, immune network, and metabolic pathways. Furthermore, the proximal genes were found to be related to germ cell and early embryo development. In summary, this study proposes several feasible SNPs and genes associated with SCK through GWAS and GSEA. These candidates can be utilized in selective breeding programs to reduce the genetic risk for SCK and subfertility in high-performance dairy cows.

Genetic relationships between behavioral traits and feed efficiency traits in lactating Holstein cows.

The evaluation of dairy cow feed efficiency using residual feed intake accounts for known energy sinks. However, behavioral traits may also contribute to the variation in feed efficiency. Our objective was to estimate the heritability and repeatability of behavioral traits and their genetic correlations with feed efficiency and its components in lactating Holstein cows. The first data set consisted of 36,075 daily rumination and lying time records collected using a SMARTBOW ear tag accelerometer (Zoetis, Parsippany, NJ) and 6,371 weekly feed efficiency records of 728 cows from the University of Wisconsin-Madison. The second data set consisted of 59,155 daily activity records, measured as number of steps, recorded by pedometers (AfiAct; S.A.E. Afikim, Kibbutz Afikim, Israel), and 8,626 weekly feed efficiency records of 635 cows from the University of Florida. Feed efficiency and its components included dry matter intake, change in body weight, metabolic body weight, secreted milk energy, and residual feed intake. The statistical models included the fixed effect of cohort, lactation number, and days in milk, and the random effects of animal and permanent environment. Heritability estimates for behavioral traits using daily records were 0.19 ± 0.06 for rumination and activity, and 0.37 ± 0.07 for lying time. Repeatability estimates for behavioral traits using daily data ranged from 0.56 ± 0.02 for activity to 0.62 ± 0.01 for lying time. Both heritability and repeatability estimates were larger when weekly records instead of daily records were used. Rumination and activity had positive genetic correlations with residual feed intake (0.40 ± 0.19 and 0.31 ± 0.22, respectively) while lying time had a negative genetic correlation with this residual feed intake (-0.27 ± 0.11). These results indicate that more efficient cows tend to spend more time lying and less time active. Additionally, less efficient cows tend to eat more and therefore also tend to ruminate longer. Overall, sensor-based behavioral traits are heritable and genetically correlated with feed efficiency and its components and, therefore, they could be used as indicators to identify feed efficient cows within the herd.

Dry matter intake in US Holstein cows: exploring the genomic and phenotypic impact of milk components and body weight composite.

Large data sets allow estimating feed required for individual milk components or body maintenance. Phenotypic regressions are useful for nutrition management, but genetic regressions are more useful in breeding programs. Dry matter intake (DMI) records from 8,513 lactations of 6,621 Holstein cows were predicted from phenotypes or genomic evaluations for milk components and body size traits. The mixed models also included days in milk, age-parity subclass, trial date, management group, and body weight change during 28- and 42-d feeding trials in mid-lactation. Phenotypic regressions of DMI on milk (0.014 ± 0.006), fat (3.06 ± 0.01), and protein (4.79 ± 0.25) were much less than corresponding genomic regressions (0.08 ± 0.03, 11.30 ± 0.47, and 9.35 ± 0.87) or sire genomic regressions multiplied by 2 (0.048 ± 0.04, 6.73 ± 0.94, and 4.98 ± 1.75). Thus, marginal feed costs as fractions of marginal milk revenue were higher from genetic than phenotypic regressions. According to the energy-corrected milk formula, fat production requires 69% more DMI than protein production. In the phenotypic regression, it was estimated that protein production requires 56% more DMI than fat. However, the genomic regression for the animal showed a difference of only 21% more DMI for protein compared with fat, while the sire genomic regressions indicated approximately 35% more DMI for fat than protein. Estimates of annual maintenance in kg DMI / kg body weight/lactation were similar from phenotypic regression (5.9 ± 0.14), genomic regression (5.8 ± 0.31), and sire genomic regression multiplied by 2 (5.3 ± 0.55) and are larger than those estimated by NASEM (2021) based on NEL equations. Multiple regressions on genomic evaluations for the 5 type traits in body weight composite (BWC) showed that strength was the type trait most associated with body weight and DMI, agreeing with the current BWC formula, whereas other traits were less useful predictors, especially for DMI. The Net Merit formula used to weight different genetic traits to achieve an economically optimal overall selection response was revised in 2021 to better account for these estimated regressions. To improve profitability, breeding programs should select smaller cows with negative residual feed intake that produce more milk, fat, and protein.

Behavioral consistency of competitive behaviors and feeding patterns in lactating dairy cows across stocking densities at the feed bunk.

Introduction: High feed bunk stocking densities can differentially impact individual dairy cows' competitive behaviors, feeding patterns, and feed efficiency. Our objective was to manipulate feed bunk stocking densities to evaluate intra-individual behavioral consistency across stocking densities and quantify associations with feed efficiency and production. Methods: Thirty-two primiparous (130.7 ± 29.0 days in milk, DIM) and 32 multiparous (111.3 ± 38.3 DIM) lactating Holstein cows were housed with 32 roughage intake control (RIC) bins. Each cow was assigned to share 8 bins with others of the same parity and similar body weight (16 cows/block; 2:1 feed bunk stocking density except during tests). Competition and feeding patterns were evaluated via video and RIC data, respectively, at 3 stocking densities (1:1, 2:1, 4:1 cows/bin) during 1-h tests (2 tests/stocking density; 6 tests/cow) following 2 h feed deprivation. Residual feed intake (RFI) was calculated across the 45-d study as the difference between observed and predicted dry matter intake (DMI) after accounting for known energy sinks. Linear mixed models were used to evaluate the overall impact of test stocking density on competition and feeding patterns. To evaluate intra-individual consistency between stocking densities, individual stability statistic (ISS) scores were computed. Correlational relationships were determined between RFI and ISS scores. Results and dicsussion: Cows displayed the most competitive behaviors at 2:1 stocking density (p < 0.0001) but experienced the highest rate of contacts per minute of eating time at 4:1 (1:1 vs. 2:1 vs. 4:1: 0.09 vs. 0.95 vs. 1.60 contacts/min; p < 0.0001). Feeding patterns were modulated as stocking density increased; eating rate increased (0.16 vs. 0.18 vs. 0.22 kg/min; p < 0.001) as eating time (40.3 vs. 28.2 vs. 14.6 min; p < 0.001) and DMI decreased (6.3 vs. 5.1 vs. 3.0 kg; p < 0.001). As stocking density doubled, individuals remained consistent (p = 0.018) in time spent near others actively eating and tended to remain consistent in competition behavior and feeding patterns (0.053 ≤ p ≤ 0.094). Between 2:1 and 4:1, cows with higher DMI and milk production were more consistent in first-visit DMI and duration. Feed efficiency was not associated with behavioral consistency across the tests (p ≥ 0.14). Nonetheless, feed bunk stocking density has behavioral implications which should be considered in nutritional management decisions.

Near-infrared spectroscopy analysis of blood plasma for predicting nonesterified fatty acid concentrations in dairy cows.

During the transition period, dairy cows are often exposed to negative energy balance (NEB), leading to lipid mobilization from adipose tissue into nonesterified fatty acids (NEFA), a common indicator of heightened illness risk. This study aimed to use blood near-infrared (NIR) spectra data to classify NEB into high or low categories, based on early-lactation cow NEFA thresholds. We collected a total of 186 plasma samples from 100 Holstein cows. The samples were categorized into critical thresholds, based on previous literature, of ≥0.60 and ≥0.70 mEq/L for identifying high NEB. Spectral data were preprocessed before the development of the predictive modes, which included the implementation of multiplicative scatter correction, standard normal variate (SNV), and first and second derivatives. The classification was performed using partial least square discriminant analyses (PLS-DA), and predictive performance was assessed using leave-one-out cross-validation. Predictive quality for each class was evaluated through specificity, precision, sensitivity, and F1 score. The study showed promising results, with the SNV technique achieving higher F1 scores. The model found 72.7% specificity, 78.9% precision, 80.8% sensitivity, and 79.8% F1 score to classify animals with NEFA levels of ≥0.60 mEq/L, and 82.1% specificity, 78.7% precision, 80.8% sensitivity, and 79.7% F1 score to classify animals with NEFA levels ≥0.70 mEq/L. These results indicate that NIR spectroscopy could serve as a tool for detecting cows under severe NEB, also showing potential for broader application across the entire transition period, as the spectral signal carried relevant information regarding cow metabolism. Furthermore, the combination of predictors derived from plasma spectra and other cow-level information can lead to more accurate disease alerts, given their relationship with the NEB.

Validation of an on-farm portable blood analyzer for quantifying blood analytes in dairy cows.

The periparturient period for dairy cows is a metabolically dynamic time period where the cow is adjusting from gestation to the onset of lactation. Metabolic disorders such as ketosis, hypocalcemia, and fatty liver occur during this time; however, tools to diagnose these diseases on-farm is limited. The need for compact metabolite quantification devices that can quantify metabolites on farm from whole blood samples is warranted. The purpose of this study was to validate a portable blood analyzer (PBA) by analyzing metabolites on privately owned dairy farms in southcentral Wisconsin. Additional tests were completed to determine if plasma metabolite quantification was similar to whole-blood quantification. Two phases were conducted on two separate farms to complete these analyses and data were analyzed by Bland-Altman plot and correlations. Metabolites quantified from whole blood samples included albumin, alanine and aspartate aminotransferases, ß-hydroxybutyrate, blood urea nitrogen, total calcium, cholesterol, creatinine kinase, γ-glutamyl transferase, glucose, magnesium, nonesterified fatty acids, phosphorous, and total protein and were analyzed in the lab after plasma separation to determine gold-standard laboratory concentrations. Across Phase 1 and 2, whole-blood PBA metabolite concentrations resulted in similar results compared to the laboratory assays. For plasma analyzed on the PBA, overall results were positively correlated, but robustness was dependent upon initial validation results indicating some metabolites are suitable for plasma quantification on the device. These results indicate that the PBA is a viable on-farm metabolite quantification tool that will be valuable for on-farm diagnosis of metabolic stress and dysfunction in transition dairy cows.

Preference for feed bins shared with cows of the same or different parity and relationships with feeding behavior and feed efficiency.

Our objectives were to (1) evaluate cows' preferences for visiting feed bins limited to either same- versus mixed-parity social interactions, depending on their parity; (2) examine the effect of parity and bin social dynamic type on competition behavior and feeding patterns, and (3) investigate cow-level relationships between feed bunk competition behavior, feeding patterns, and feed efficiency. Twenty-eight primiparous and 28 multiparous (2.4 ± 0.6 lactations) lactating Holstein cows (127.8 ± 30.1 and 145.3. ± 10.4 DIM, respectively) were housed in a freestall pen with 28 roughage intake control bins (2:1 stocking density). Each cow was assigned to 2 bins, including 1 shared with 3 other cows of the same parity (SM) and 1 with 3 cows of mixed parities (MX, 50% primiparous and 50% multiparous). Feed bunk competition was recorded via video in the first hour after morning feed delivery for 2 d, and feeding patterns were recorded from 24-h roughage intake control data. Residual feed intake was calculated as the difference between predicted and observed dry matter intake after accounting for known energy sinks. Based on the first visit to the feed bunk after fresh feed delivery, multiparous cows tended to prefer the MX bin compared with the SM one; cows showed no other overall preference for bin type based on number of visits. Over time, multiparous cows remained consistent in their magnitude of preference for visiting each bin type, but involvement in competition was not consistent over time. Primiparous cows tended to be involved in more total competitive contacts and ate faster at the SM bin compared with the MX one. Those primiparous cows who visited the SM bin more often within the first hour after morning feed delivery tended to be less feed efficient. Multiparous cows initiated more successful replacements after a displacement at the MX versus SM bin, with no difference in feeding patterns between bin types. Regardless of parity or bin type, visiting the bunk sooner after feed delivery was correlated with involvement in more competitive interactions and more time eating within the first 30 min. Consuming more feed during a longer first visit to the bunk after fresh feed delivery was correlated with being less feed efficient. Overall, when given the choice of feeding from bins shared with cows of the same or mixed parities at a 2:1 stocking density, primiparous cows showed differences in behavior between those bin types, with implications for feed efficiency; these effects are perhaps an unintended consequence of compensatory strategies to avoid direct competition with multiparous cows.

Consistency of dry matter intake in Holstein cows: Heritability estimates and associations with feed efficiency.

Resilience can be defined as the capacity to maintain performance or bounce back to normal functioning after a perturbation, and studying fluctuations in daily feed intake may be an effective way to identify resilient dairy cows. Our goal was to develop new phenotypes based on daily dry matter intake (DMI) consistency in Holstein cows, estimate genetic parameters and genetic correlations with feed efficiency and milk yield consistency, and evaluate their relationships with production, longevity, health, and reproduction traits. Data consisted of 397,334 daily DMI records of 6,238 lactating Holstein cows collected from 2007 to 2022 at 6 research stations across the United States. Consistency phenotypes were calculated based on the deviations from expected daily DMI for individual cows during their respective feeding trials, which ranged from 27 to 151 d in duration. Expected values were derived from different models, including simple average, quadratic and cubic quantile regression with a 0.5 quantile, and locally estimated scatterplot smoothing (LOESS) regression with span parameters 0.5 and 0.7. We then calculated the log of variance (log-Var-DMI) of daily deviations for each model as the consistency phenotype. Consistency of milk yield was also calculated, as a reference, using the same methods (log-Var-Milk). Genetic parameters were estimated using an animal model, including lactation, days in milk and cohort as fixed effects, and animal as random effect. Relationships between log-Var-DMI and traits currently considered in the US national genetic evaluation were evaluated using Spearman's rank correlations between sires' breeding values. Heritability estimates for log-Var-DMI ranged from 0.11 ± 0.02 to 0.14 ± 0.02 across models. Different methods (simple average, quantile regressions, and LOESS regressions) used to calculate log-Var-DMI yielded very similar results, with genetic correlations ranging from 0.94 to 0.99. Estimated genetic correlations between log-Var-DMI and log-Var-Milk ranged from 0.51 to 0.62. Estimated genetic correlations between log-Var-DMI and feed efficiency ranged from 0.55 to 0.60 with secreted milk energy, from 0.59 to 0.63 with metabolic body weight, and from 0.26 to 0.31 with residual feed intake (RFI). Relationships between log-Var-DMI and the traits in the national genetic evaluation were moderate and positive correlations with milk yield (0.20 to 0.21), moderate and negative correlations with female fertility (-0.07 to -0.20), no significant correlations with health and longevity, and favorable correlations with feed efficiency (-0.23 to -0.25 with feed saved and 0.21 to 0.26 with RFI). We concluded that DMI consistency is heritable and may be an indicator of resilience. Cows with lower variation in the difference between actual and expected daily DMI (more consistency) may be more effective in maintaining performance in the face of challenges or perturbations, whereas cows with greater variation in observed versus expected daily DMI (less consistency) are less feed efficient and may be less resilient.

Liver and Muscle Transcriptomes Differ in Mid-Lactation Cows Divergent in Feed Efficiency in the Presence or Absence of Supplemental Rumen-Protected Choline.

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 ß-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.

Social interactions, feeding patterns, and feed efficiency of same- and mixed-parity groups of lactating cows.

Social dynamics in group-housed animals can have important effects on their welfare, feed efficiency, and production potential. Our objectives were to: (1) evaluate the effects of parity and social grouping on competition behavior, feeding patterns, and feed efficiency, and (2) investigate cow-level relationships between competition and feeding behavior, production, and feed efficiency. Fifty-nine Holstein cows (144.5 ± 21.8 starting days in milk, mean ± SD) were housed in a freestall pen with 30 Roughage Intake Control (RIC) bins. We evaluated the effects of parity (primiparous [PR, n = 29] vs. multiparous [MU, n = 30]) and group composition at the feed bunk (same-parity [SM, n = 39] vs. mixed-parity [MX, n = 20, 50% of each parity]) with a 2 × 2 factorial design (SM-MU: n = 20; SM-PR: n = 19; MX-MU: n = 10; MX-PR: n = 10) on competition behavior, feeding patterns, and feed efficiency. Within the pen, groups of 9 to 10 cows were considered subgroups and assigned to treatments defined by sets of 5 assigned bins (2:1 stocking density). Feed bunk competition and feeding patterns were recorded via continuous video in the first hour after morning feed delivery and 24-h RIC data, respectively. Residual feed intake (RFI) was calculated as the difference between predicted and observed dry matter intake (DMI) after accounting for known energy sinks. Linear models were used to evaluate the effects and interactions of parity and group composition on competition, feeding behavior, and feed efficiency. Within-cow correlations were performed between competition, feeding behavior, and RFI. Cows in MX, compared with SM, were involved in more competitive interactions [mean (95% CI): competitive contacts: 11.5 (8.1, 16.3) vs. 7.2 (5.5, 9.3) events; displacements: 4.0 (3.0, 5.3) vs. 2.1 (1.7, 2.7) events, and replacements: 3.5 (2.6, 4.7) vs. 1.9 (1.5, 2.5) events]. Cows in MX vs. those in SM had more bunk visits/meal ( 4.3 [3.9, 4.8] vs. 3.7 [3.4, 3.9] visits/meal) and longer meals (31.2 vs. 27.4 ± 0.9 min/meal) and tended to have higher RFI (0.41 ± 0.3 vs. -0.21 ± 0.2) and were therefore less feed efficient. Multiparous versus PR cows had greater DMI per day (29.3 ± 0.6 vs. 25.5 ± 0.4 kg/d) and per meal (4.2 [4.0, 4.4] vs. 3.4 [3.2, 3.6] kg/meal), faster eating rates (0.14 [0.13, 0.15] vs. 0.12 [0.11, 0.13] kg/min), and fewer bunk visits/d (26.6 [24.0, 29.4] vs. 32.8 [29.7, 35.9]). Regardless of grouping or parity, cows with shorter latencies to first visit the bunk after feed delivery were involved in more competition and tended to be less feed efficient. Overall, individual cow- and group-level relationships among competition, feeding behavior, and feed efficiency play an important role in feed bunk social dynamics. At a competitive 2:1 stocking density, mixed-parity groups for lactating cows may have potentially negative animal welfare and feed efficiency implications that should be considered when selecting grouping strategies on the farm.

In utero choline exposure alters growth, metabolism, feed efficiency, and carcass characteristics of Holstein × Angus cattle from weaning to slaughter.

Feeding rumen-protected choline (RPC) to late gestation dairy cows has potential to affect growth in offspring. The objective of this study was to evaluate the effects of in utero choline exposure on the growth, feed efficiency (FE), metabolism, and carcass quality of Angus × Holstein cattle. Multiparous Holstein cows pregnant with male (N = 17) or female (N = 30) Angus-sired calves were enrolled 21 d prepartum and randomly assigned to one of four dietary treatments varying in quantity and formulation of RPC. The treatments included a control with 0 g/d supplemental RPC (CTL), supplemental RPC fed at the recommended dose (RD) of 15 g/d from either an established RPC product (RPC1RD; ReaShure; Balchem Corp.) or choline ion from a concentrated RPC prototype (RPC2RD; Balchem Corp.), or a high dose (HD) of RPC2 fed at 22 g/d (RPC2HD). From 2 to 6 mo of age, calves were group housed and offered 2.3 kg grain/hd/d (42% CP) with ad libitum grass hay, and stepped up to a complete finishing diet by 7 mo (12.0% CP; 1.34 Mcal/kg NEg). Weight and height were measured monthly. Animal FE was measured in individual pens for 35 d at 8 mo. Feed intake was measured daily, and blood was obtained on day 18 during the FE period. Afterwards, cattle were group housed and offered a free-choice finishing diet until slaughter, where carcass yield and quality characteristics were measured. Mixed models were used in PROC MIXED (SAS, 9.4) with the fixed effects of treatment, sex, time, their interactions, and the random effect of calf. Month was the repeated measure, and preplanned contrasts were used. Blood and FE data were analyzed with the fixed effect of dam choline treatment, calf sex, and the interaction. Increasing dose of RPC tended to increase weight over the entire study period. Feeding any RPC increased hip and wither height compared with CTL, and increasing RPC dose linearly increased hip and wither height. Treatment and sex interacted on DMI whereby increasing RPC intake linearly increased DMI for males but not females. Compared with control, feeding any RPC decreased plasma insulin, glucose, and an insulin sensitivity index (RQUICKI). In utero choline exposure increased kidney-pelvic-heart fat and marbling score. Mechanisms of action for intrauterine choline exposure on offspring growth, metabolism, and carcass characteristics should be explored as they have direct implications for profitability for cattle growers and feeders.

Common nutritional and management programs implemented during gestation in dairy cattle also have positive outcomes for offspring growth, health, and well-being. Recent work has demonstrated that supplementing rumen-protected choline (RPC) to dairy cows for several weeks before calving increases growth and feed efficiency (FE) of their calves. Considering the recent industry trends of breeding dairy cows with beef semen, any potential growth and FE advantages imparted by prenatal RPC supplementation of the dams could help increase value of the resulting beef × dairy calves. The objective of this study was to evaluate growth, FE, and carcass characteristics of beef × dairy calves from dairy cows supplemented with RPC before calving. Feeding RPC to dairy cows before calving increased offspring weight and height through 9 mo of age. In utero exposure to choline also improved markers of insulin sensitivity of the offspring while being fed with a high-energy diet. Dam dietary RPC supplementation increased offspring kidney, pelvic, and heart fat at slaughter, and also increased marbling score. Considering the importance of marbling in carcass quality, the potential of RPC to positively influence offspring performance could be beneficial for further enhancing value of beef × dairy cattle at slaughter.

Impact of parity differences on residual feed intake estimation in Holstein cows.

Residual feed intake (RFI) has been used as a measure of feed efficiency in farm animals. In lactating dairy cattle, RFI is typically obtained as the difference between dry matter intake observations and predictions from regression on known energy sinks, and effects of parity, days in milk, and cohort. The impact of parity (lactation number) on the estimation of RFI is not well understood, so the objectives of this study were to (1) evaluate alternative RFI models in which the energy sinks (metabolic body weight, body weight change, and secreted milk energy) were nested or not nested within parity, and (2) estimate variance components and genetic correlations for RFI across parities. Data consisted of 72,474 weekly RFI records of 5,813 lactating Holstein cows collected from 2007 to 2022 in 5 research stations across the United States. Estimates of heritability, repeatability, and genetic correlations between weekly RFI for parities 1, 2, and 3 were obtained using bivariate repeatability animal models. The nested RFI model showed better goodness of fit than the nonnested model, and some partial regression coefficients of dry matter intake on energy sinks were heterogeneous between parities. However, the Spearman's rank correlation between RFI values calculated from nested and nonnested models was equal to 0.99. Similarly, Spearman's rank correlation between the RFI breeding values from these 2 models was equal to 0.98. Heritability estimates for RFI were equal to 0.16 for parity 1, 0.19 for parity 2, and 0.22 for parity 3. Repeatability estimates for RFI across weeks within parities were high, ranging from 0.51 to 0.57. Spearman's rank correlations of sires' breeding values were 0.99 between parities 1 and 2, 0.91 between parities 1 and 3, and 0.92 between parities 2 and 3. We conclude that nesting energy sinks within parity when computing RFI improves model goodness of fit, but the impact on the estimated breading values appears to be minimal.

Dam Body Condition Score Alters Offspring Circulating Cortisol and Energy Metabolites in Holstein Calves but Did Not Affect Neonatal Leptin Surge.

The neonatal leptin surge is important for hypothalamic development, feed intake regulation, and long-term metabolic control. In sheep, the leptin surge is eliminated with maternal overnutrition and an elevated dam body condition score (BCS), but this has not been assessed in dairy cattle. The aim of this study was to characterize the neonatal profile of leptin, cortisol and other key metabolites in calves born to Holstein cows with a range of BCS. Dam BCS was determined 21 d before expected parturition. Blood was collected from calves within 4 h of birth (d 0), and on days 1, 3, 5, and 7. Serum was analyzed for concentrations of leptin, cortisol, blood urea nitrogen, ß-hydroxybutyrate (BHB), free fatty acids (FFA), triglycerides, and total protein (TP). Statistical analysis was performed separately for calves sired by Holstein (HOL) or Angus (HOL-ANG) bulls. Leptin tended to decrease after birth in HOL calves, but there was no evidence of an association between leptin and BCS. For HOL calves, the cortisol level increased with an increasing dam BCS on day 0 only. Dam BCS was variably associated with the calf BHB and TP levels, depending on the sire breed and day of age. Further investigation is required to elucidate the impacts of maternal dietary and energy status during gestation on offspring metabolism and performance, in addition to the potential impact of the absence of a leptin surge on long-term feed intake regulation in dairy cattle.

Impact of epistasis effects on the accuracy of predicting phenotypic values of residual feed intake in U. S Holstein cows.

The impact of genomic epistasis effects on the accuracy of predicting the phenotypic values of residual feed intake (RFI) in U.S. Holstein cows was evaluated using 6215 Holstein cows and 78,964 SNPs. Two SNP models and seven epistasis models were initially evaluated. Heritability estimates and the accuracy of predicting the RFI phenotypic values from 10-fold cross-validation studies identified the model with SNP additive effects and additive × additive (A×A) epistasis effects (A + A×A model) to be the best prediction model. Under the A + A×A model, additive heritability was 0.141, and A×A heritability was 0.263 that consisted of 0.260 inter-chromosome A×A heritability and 0.003 intra-chromosome A×A heritability, showing that inter-chromosome A×A effects were responsible for the accuracy increases due to A×A. Under the SNP additive model (A-only model), the additive heritability was 0.171. In the 10 validation populations, the average accuracy for predicting the RFI phenotypic values was 0.246 (with range 0.197-0.333) under A + A×A model and was 0.231 (with range of 0.188-0.319) under the A-only model. The average increase in the accuracy of predicting the RFI phenotypic values by the A + A×A model over the A-only model was 6.49% (with range of 3.02-14.29%). Results in this study showed A×A epistasis effects had a positive impact on the accuracy of predicting the RFI phenotypic values when combined with additive effects in the prediction model.

Prediction of Liver Triglyceride Content in Early Lactation Multiparous Holstein Cows Using Blood Metabolite, Mineral, and Protein Biomarker Concentrations.

Bovine fatty liver syndrome (bFLS) is difficult to diagnose because a liver tissue biopsy is required to assess liver triglyceride (TG) content. We hypothesized that a blood biomarker panel could be a convenient alternative method of liver TG content assessment and bFLS diagnosis. Our objectives were to predict liver TG using blood biomarker concentrations across days in milk (DIM; longitudinal, LT) or at a single timepoint (ST; 3, 7, or 14 DIM), as well as different biomarker combination based on their perceived accessibility. Data from two separate experiments (n = 65 cows) was used for model training and validation. Response variables were based on the maximum liver TG observed in 1 and 14 DIM liver biopsies: Max TG (continuous), Low TG (TG > 13.3% dry matter; DM), Median TG (TG > 17.1% DM), and High TG (TG > 22.0% DM). Model performance varied but High TG was well predicted by sparse partial least squares­discriminate analysis models using LT and ST data, achieving balanced error rates ≤ 15.4% for several model variations during cross-validation. In conclusion, blood biomarker panels using 7 DIM, 14 DIM, or LT data may be a useful diagnostic tool for bFLS in research and field settings.

Estimates of genetic parameters for feeding behavior traits and their associations with feed efficiency in Holstein cows.

Residual feed intake (RFI) is commonly used to measure feed efficiency but individual intake recording systems are needed. Feeding behavior may be used as an indicator trait for feed efficiency using less expensive precision livestock farming technologies. Our goal was to estimate genetic parameters for feeding behavior and the genetic correlations with feed efficiency in Holstein cows. Data consisted of 75,877 daily feeding behavior records of 1,328 mid-lactation Holstein cows in 31 experiments conducted from 2009 to 2020 with an automated intake recording system. Feeding behavior traits included number of feeder visits per day, number of meals per day, duration of each feeder visit, duration of each meal, total duration of feeder visits, intake per visit, intake per meal [kg of dry matter (DM)], feeding rate per visit, and feeding rate per meal (kg of DM per min). The meal criterion was estimated as 26.4 min, which means that any pair of feeder visits separated by less than 26.4 min were considered part of the same meal. The statistical model included lactation and days in milk as fixed effects, and experiment-treatment, animal, and permanent environment as random effects. Genetic parameters for feeding behavior traits were estimated using daily records and weekly averages. Estimates of heritability for daily feeding behavior traits ranged from 0.09 ± 0.02 (number of meals; mean ± standard error) to 0.23 ± 0.03 (feeding rate per meal), with repeatability estimates ranging from 0.23 ± 0.01 (number of meals) to 0.52 ± 0.02 (number of feeder visits). Estimates of heritability for weekly averages of feeding behavior traits ranged from 0.19 ± 0.04 (number of meals) to 0.32 ± 0.04 (feeding rate per visit), with repeatability estimates ranging from 0.46 ± 0.02 (duration of each meal) to 0.62 ± 0.02 (feeding rate per visit and per meal). Most of the feeding behavior measures were strongly genetically correlated, showing that with more visits or meals per day, cows spend less time in each feeder visit or meal with lower intake per visit or meal. Weekly averages for feeding behavior traits were analyzed jointly with RFI and its components. Number of meals was genetically correlated with milk energy (0.48), metabolic body weight (-0.27), and RFI (0.19). Duration of each feeder visit and meal were genetically correlated with milk energy (0.43 and 0.44, respectively). Total duration of feeder visits per day was genetically correlated with DM intake (0.29), milk energy (0.62), metabolic body weight (-0.37), and RFI (0.20). Intake per visit and meal were genetically correlated with DM intake (0.63 and 0.87), milk energy (0.47 and 0.69), metabolic body weight (0.47 and 0.68), and RFI (0.31 and 0.65). Feeding rate was genetically correlated with DM intake (0.69), metabolic body weight (0.67), RFI (0.47), and milk energy (0.21). We conclude that measures of feeding behavior could be useful indicators of dairy cow feed efficiency, and individual cows that eat at a slower rate may be more feed efficient.

Responsiveness of PNPLA3 and lipid-related transcription factors is dependent upon fatty acid profile in primary bovine hepatocytes.

Knockdown of patatin-like phospholipase domain-containing protein 3 (PNPLA3) increased triglycerides (TG) in primary bovine hepatocytes, suggesting that PNPLA3 plays a causal role in hepatic TG clearing. In vivo, PNPLA3 abundance across the periparturient period is inversely related to hepatic TG accumulation and circulating fatty acid (FA) concentrations. The purpose of this research was to determine if PNPLA3, as well as other lipases, transcription factors, or FA-mediated genes, are regulated by FA mimicking liver lipid accumulation (ACCUM) and liver lipid clearing (RECOV) or singular FA physiologically found in dairy cows at 0.5 mM of circulating RECOV (iRECOV). Abundance of PNPLA3 tended to decrease with ACCUM and increased quadratically with RECOV (P ≤ 0.10), differing from PNPLA3 expression, but consistent with previous in vivo research. Adipose TG lipase abundance, but not other lipase abundances, was quadratically responsive to both ACCUM and RECOV (P ≤ 0.005). Abundance of PNPLA3 and SREBP1c and expression of LXRA responded similarly to iRECOV, with C18:0 tending to decrease abundance (P ≤ 0.07). Results indicate that bovine PNPLA3 is translationally regulated by FA and although a LXRA-SREBP1c pathway mediation is possible, the mechanism warrants further investigation.

Circulating Metabolites Indicate Differences in High and Low Residual Feed Intake Holstein Dairy Cows.

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.

Novel Facets of the Liver Transcriptome Are Associated with the Susceptibility and Resistance to Lipid-Related Metabolic Disorders in Periparturient Holstein Cows.

Lipid-related metabolic disorders (LRMD) are prevalent in early lactation dairy cows, and have detrimental effects on productivity and health. Our objectives were to identify cows resistant or susceptible to LRMD using a ketosis induction protocol (KIP) to discover differentially expressed liver genes and metabolic pathways associated with disposition. Clustering cows based on postpartum lipid metabolite concentrations within dietary treatments identified cows more or less susceptible (MS vs. LS) to LRMD within the control treatment, and more or less resistant (MR vs. LR) within the KIP treatment. Whole-transcriptome RNA sequencing was performed on liver samples (-28, +1, and +14 days relative to calving) to assess differential gene and pathway expression (LS vs. MS; MR vs. LR; n = 3 cows per cluster). Cows within the MS and LR clusters had evidence of greater blood serum ß-hydroxybutyrate concentration and liver triglyceride content than the LS and MR clusters, respectively. The inferred metabolism of differentially expressed genes suggested a role of immune response (i.e., interferon-inducible proteins and major histocompatibility complex molecules). Additionally, unique roles for glutathione metabolism and eicosanoid metabolism in modulating susceptibility and resistance, respectively, were implicated. Overall, this research provides novel insight into the role of immunometabolism in LRMD pathology, and suggests the potential for unique control points for LRMD progression and severity.

Assessment of the Relationship between Postpartum Health and Mid-Lactation Performance, Behavior, and Feed Efficiency in Holstein Dairy Cows.

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.