It is now widely accepted that dairy cow performance is influenced by both the host genome and rumen microbiome composition. The contributions of the genome and the microbiome to the phenotypes of interest are quantified by heritability (h²) and microbiability (m²), respectively. However, if the genome and microbiome are included in the model, then the h² reflects only the contribution of the direct genetic effects quantified as direct heritability (h_d²), and the holobiont effect reflects the joint action of the genome and the microbiome, quantified as the holobiability (ho²). The objectives of this study were to estimate h², h_d²,m², and ho² for dry matter intake, milk energy, and residual feed intake; and to evaluate the predictive ability of different models, including genome, microbiome, and their interaction. Data consisted of feed efficiency records, SNP genotype data, and 16S rRNA rumen microbial abundances from 448 mid-lactation Holstein cows from 2 research farms. Three kernel models were fit to each trait: one with only the genomic effect (model G), one with the genomic and microbiome effects (model GM), and one with the genomic, microbiome, and interaction effects (model GMO). The model GMO, or holobiont model, showed the best goodness-of-fit. The h_d² estimates were always 10% to 15% lower than h² estimates for all traits, suggesting a mediated genetic effect through the rumen microbiome, and m² estimates were moderate for all traits, and up to 26% for milk energy. The ho² was greater than the sum of h_d² and m², suggesting that the genome-by-microbiome interaction had a sizable effect on feed efficiency. Kernel models fitting the rumen microbiome (i.e., models GM and GMO) showed larger predictive correlations and smaller prediction bias than the model G. These findings reveal a moderate contribution of the rumen microbiome to feed efficiency traits in lactating Holstein cows and strongly suggest that the rumen microbiome mediates part of the host genetic effect.

Objectives were to assess reproductive and productive outcomes associated with failure of clinical cure in dairy cows diagnosed with metritis following antimicrobial therapy. This retrospective cohort study included data from 3 experiments performed in 5 dairies. Metritis was characterized by the presence of watery, fetid, reddish-brownish vaginal discharge within 21 DIM (study d 0). Cows not diagnosed with metritis (i.e., cows may have had other diseases postpartum; NoMT; n = 1,194) were paired based on lactation number and calving date. All cows with metritis received antimicrobial therapy (ampicillin or ceftiofur). Clinical cure was evaluated on d 10 based on vaginal discharge score, and cows were categorized as cured (MTC; n = 1,111) or not cured (MTnoC; n = 299). Purulent vaginal discharge (28 ± 3 or 32 ± 3 DIM), cytological endometritis (35 ± 3 or 39 ± 3 DIM), and estrous cyclicity (50 ± 3 and 64 ± 3, 36 ± 3 and 50 ± 3, or 37 ± 5 and 51 ± 5 DIM) were evaluated in subgroups of cows. Proportions of cows with purulent vaginal discharge and cytological endometritis were greatest for MTnoC (91.7 and 91.4%), intermediate for MTC (74.0 and 73.3%), and smallest for NoMT (38.1 and 36.4%). Proportion of cyclic cows was smaller for MTnoC compared with MTC and NoMT (62.0, 71.0, and 71.0%). Pregnancy per artificial insemination following first service was smaller for cows with metritis compared with their counterparts with no metritis (NoMT = 28.1, MTC = 26.1, MTnoC = 22.0%). Pregnancy loss tended to be greater for MTnoC compared with MTC (NoMT = 11.5, MTC = 11.1, MTnoC = 18.4%). Hazard of pregnancy by 300 DIM was smallest for MTnoC, intermediate for MTC, and greatest for NoMT. Death by 60 DIM (3.9, 1.1, and 0.6%) and removal from herd by 300 DIM (26.3, 17.4, and 15.4%) were greatest for MTnoC compared with MTC and NoMT, respectively. Milk production among multiparous cows was smaller for MTnoC compared with MTC and NoMT in the first 10 mo postpartum, whereas MTC produced less milk compared with NoMT only during the first 2 mo postpartum (NoMT = 42.0 ± 0.22, MTC = 40.6 ± 0.28, MTnoC = 37.7 ± 0.54 kg/d). Failure of clinical cure was not associated with milk yield in primiparous cows (NoMT = 35.2 ± 0.31, MTC = 33.9 ± 0.31, MTnoC = 35.0 ± 0.52 kg/d). Cows diagnosed with metritis that do not undergo clinical cure by 10 d of onset of antimicrobial therapy have impaired reproductive performance, reduced milk production, and increased risk of leaving the herd.

Our objectives were to compare the effects of ceftiofur crystalline free acid (CCFA) and ampicillin trihydrate (AMP) treatments of cows diagnosed with metritis on uterine health, behavior, reproductive, and productive responses. A controlled randomized clinical trial was designed. Metritis was defined as vaginal discharge (VD) = 5 (fetid, watery, red/brown) within 21 d in milk (DIM) and rectal temperature (RT) <39.5°C, whereas VD = 5 and RT ≥39.5°C was defined as puerperal metritis. On the day of diagnosis (d 0), cows were paired by parity and severity of metritis (metritis vs. puerperal metritis) and assigned randomly to the AMP and CCFA treatments. Cows enrolled in the AMP (n = 308) treatment were moved to a nonsalable-milk pen, where they were treated once daily for 5 d, and were moved back to their original pen 72 h after the last treatment (d 7). Cows enrolled in the CCFA (n = 310) treatment remained in their original pen and received 2 treatments of CCFA, 72 h apart. Rectal temperature was measured daily from d 0 to 6 and on d 11. Vaginal discharge was evaluated on d 4, 6, and 11 to assess cure. Cure was defined as the absence of treatment with additional antimicrobial before experiment d 11, VD <5, and RT <39.5°C. Cows were examined at 28 ± 3 DIM for purulent VD (PVD) and at 35 ± 3 DIM for cytological endometritis. Pregnancy was diagnosed at 40 ± 3 and 60 ± 7 d after first and second artificial inseminations. Cure of metritis did not differ between treatments on d 11 (AMP = 64.6 ± 3.1, CCFA = 63.5 ± 3.1%). Cows treated with AMP had greater RT from experiment d 1 to 6 compared with cows treated with CCFA (AMP = 39.1 ± 0.02, CCFA = 39.0 ± 0.02°C). Cows in the AMP treatment had greater prevalence of PVD at 28 ± 3 DIM (AMP = 82.6 ± 2.3, CCFA = 74.4 ± 2.7%) and tended to have greater prevalence of cytological endometritis at 35 ± 3 DIM (AMP = 77.8 ± 6.2 vs. CCFA = 61.7 ± 7.5%) than CCFA-treated cows. Treatment did not affect the hazard of pregnancy among multiparous cows; however, among primiparous cows, CCFA treatment reduced the hazard of pregnancy and increased the median days to pregnancy (AMP = 145 vs. CCFA = 169 d). Finally, average daily milk yield up to 14 wk postpartum was not affected by treatment (AMP = 38.0 ± 0.4, CCFA = 37.5 ± 0.4 kg). We conclude from the current experiment that CCFA was more effective in reducing RT and improving uterine health of metritic cows; however, the improved hazard of pregnancy of primiparous cows treated with AMP is important and warrants further investigation.

Objectives were to evaluate differences in the uterine and serum metabolomes associated with metritis in dairy cows. Vaginal discharge was evaluated using a Metricheck device (Simcro) at 5, 7, and 11 d in milk (DIM; herd 1) or 4, 6, 8, 10, and 12 DIM (herd 2). Cows with reddish or brownish, watery, and fetid discharge were diagnosed with metritis (n = 24). Cows with metritis were paired with herdmates without metritis (i.e., clear mucous vaginal discharge or clear lochia with ≤50% of pus) based on DIM and parity (n = 24). Day of metritis diagnosis was considered study d 0. All cows diagnosed with metritis received antimicrobial therapy. The metabolome of uterine lavage collected on d 0 and 5, and serum samples collected on d 0 were evaluated using untargeted gas chromatography time-of-flight mass spectrometry. Normalized data were subjected to multivariate canonical analysis of population using the MultBiplotR and MixOmics packages in R Studio. Univariate analyses including t-test, principal component analyses, partial least squares discriminant analyses, and pathway analyses were conducted using Metaboanalyst. The uterine metabolome differed between cows with and without metritis on d 0. Differences in the uterine metabolome associated with metritis on d 0 were related to the metabolism of butanoate, amino acids (i.e., glycine, serine, threonine, alanine, aspartate, and glutamate), glycolysis and gluconeogenesis, and the tricarboxylic acid cycle. No differences in the serum metabolome were observed between cows diagnosed with metritis and counterparts without metritis on d 0. Similarly, no differences in uterine metabolome were observed between cows with metritis and counterparts not diagnosed with metritis on d 5. These results indicate that the establishment of metritis in dairy cows is associated with local disturbances in amino acid, lipid, and carbohydrate metabolism in the uterus. The lack of differences in the uterine metabolome on d 5 indicates that processes implicated with the disease are reestablished by d 5 after diagnosis and treatment.

Objectives of the current experiment were to evaluate the effects on reproductive performance of adding an automated estrous detection (AED) device to the reproductive management of Holstein cows. In addition, we aimed to determine whether the AED device would equally influence the reproductive performance of cows receiving artificial insemination and those receiving embryo transfer. Cows (n = 1,244) were observed daily for estrous detection by trained farm personnel with the aid of a tail head mounting device. Cows (n = 641) enrolled in the AED treatment were fitted with a neck-mounted automated estrus detection device, and cows enrolled in the control (CTRL, n = 603) treatment were not fitted with the AED device. Cows not detected in estrus by 54 ± 3 DIM received 1 injection of PGF2α, and those not detected in estrus by 68 ± 3 DIM were enrolled in an ovulation synchronization protocol. The hazard of first service tended to be affected by treatment [CTRL: referent, AED: adjusted hazard ratio (AHR) = 1.11, 95% confidence interval (CI) = 0.98, 1.25], and we determined a tendency for the AED treatment to reduce the proportion of cows receiving the first service at fixed time (39.7 ± 5.7 vs. 45.5 ± 5.8%). After the first service, high-producing cows enrolled in the AED treatment were more likely to be pregnant (19.1 ± 2.4 vs. 31.8 ± 3.0%), but no differences between the AED and CTRL treatments were observed among low-producing cows (24.9 ± 2.9 vs. 24.7 ± 2.9%). The hazard of second service was affected by treatment (CTRL: referent, AED: AHR = 1.22, 95% CI = 1.01, 1.47), and the AED treatment reduced the proportion of cows receiving the second service at fixed time compared with the CTRL treatment (15.7 ± 2.2 vs. 21.9 ± 2.5%). After the second service, the AED treatment tended to increase the proportion of cows pregnant (31.0 ± 3.0 vs. 24.9 ± 2.6%) and tended to reduce the proportion of cows that lost pregnancy between the first and second pregnancy exams (9.7 ± 3.3 vs. 16.7 ± 4.6%). The interaction between treatment and type of service did not affect percentage of cows pregnant and pregnancy loss to the first and second services. The hazard of pregnancy was affected by the interaction between treatment and milk yield, in that cows with above the median milk yield in the AED treatment became pregnant at a faster rate than CTRL cows, but no difference between treatments was observed among cows with milk yield below the median. Cows with above the median milk yield had shorter duration of estrus and were less likely to have activity peak at estrus ≥89 (0 = minimum, 100 = maximum). The current experiment suggests that AED devices may improve reproductive performance not only by increasing service rates but also by improving accuracy of estrous detection. According to the current experiment, high-producing cows may benefit most from the addition of AED devices to reproductive management.

The objective was to unravel the peripartum immune and metabolic changes associated with metritis in Holstein cows. Holstein cows (n = 128) had blood collected at -14, 0, 3, and 7 d relative to parturition (DRP). Flow cytometry was used to evaluate blood leukocyte counts, proportions, and activation. Total cells, live cells (LiveDead-), single cells, monocytes (CD172α⁺/CD14⁺), polymorphonuclears (PMN; CD172α⁺/CD14^-/SSC^high), B-cells (CD21⁺/MHCII⁺), CD4⁺ T-cells (CD4⁺), CD8⁺ T-cells (CD8⁺), and γδ T-cells (γδTCR⁺) were evaluated. CD62L and CD11b were used as markers of cell activation. Major histocompatibility complex class II was used as a marker of antigen presentation in monocytes. A MILLIPLEX Bovine Cytokine/Chemokine 08-plex kit was used to evaluate plasma concentrations of interferon-γ, Interleukin (IL)-1α, IL-1β, IL-4, IL-6, IL-8, IL-10, and tumor necrosis factor-α. The body weight (BW) change prepartum was calculated as the difference between calving BW and prepartum BW divided by the number of days between measurements. Plasma fatty acids (FA) were measured at -14 and 0 DRP using untargeted gas chromatography with time-of-flight mass spectrometry. Data were analyzed by ANOVA for repeated measures. Cows that developed metritis (n = 57) had greater prepartum BW, prepartum BW loss, and greater FA concentrations at calving. Plasma FA at calving was positively correlated with IL-1β. Cows that developed metritis had persistent systemic inflammation, which was demonstrated by greater B-cell activation, greater pro-inflammatory cytokine concentrations, and greater cell damage pre- and postpartum. Postpartum, we observed greater polymorphonuclear cell activation and extravasation but lesser monocytes and CD4⁺ T-cells activation and extravasation, which suggests postpartum immune tolerance. Greater prepartum adiposity in cows that developed metritis may lead to systemic inflammation pre- and postpartum and immune tolerance postpartum, which may lead to failure to prevent bacterial infection, and development of puerperal metritis.