Reproductive physiological outcomes of dairy cows with different genomic merit for fertility: biomarkers, uterine health, endocrine status, estrus features, and response to ovarian synchronization.

Our overarching objective was to characterize associations between genomic merit for fertility and the reproductive function of lactating dairy cows in a prospective cohort study. In this manuscript, we present results of the association between genomic merit for fertility and indicators of metabolic status and inflammation, uterine health, endocrine status, response to synchronization, and estrous behavior in dairy cows. Lactating Holstein cows entering their first (n = 82) or second (n = 37) lactation were enrolled at parturition and fitted with an ear-attached sensor for automated detection of estrus. Ear-notch tissue samples were collected from all cows and submitted for genotyping using a commercial genomic test. Based on genomic predicted transmitting ability values for daughter pregnancy rate (gDPR) cows were classified into a high (Hi-Fert; gDPR > 0.6; n = 36), medium (Med-Fert; gDPR -1.3 to 0.6; n = 45), and low (Lo-Fert; gDPR < -1.3; n = 38) group. At 33 to 39 d in milk (DIM), cohorts of cows were enrolled in the Presynch-Ovsynch protocol for synchronization of estrus and ovulation. Body weights, body condition scores (BCS), and uterine health measurements (i.e., vaginal discharge, uterine cytology) were collected from parturition to 60 DIM and milk yield was collected through 90 DIM. Blood samples were collected weekly through 3 wk of lactation for analysis of ß-hydroxybutyrate, nonesterified fatty acids, and haptoglobin plasma concentrations. Body weight, BCS, NEFA, BHB, and Haptoglobin were not associated with fertility groups from 1 to 9 wk after parturition. The proportion of cows classified as having endometritis at 33 to 36 DIM tended to be greater for the Lo-Fert than the Hi-Fert group. The proportion of cows that resumed cyclicity did not differ at any time point evaluated and there were no significant associations between probability or duration and intensity of estrus with fertility group. Cows of superior genetic merit for fertility were more likely to ovulate, have a functional CL, have greater circulating P4, and have larger ovulatory size than cows of inferior fertility potential at key time points during synchronization of estrus and ovulation. Despite observing numerical differences with potential performance consequences for the proportion of cows that responded to synchronization of ovulation and were both cyclic and responded to the Ovsynch portion of the synchronization protocol, we did not observe significant differences between fertility groups. Although not consistent and modest in magnitude, the collective physiological and endocrine differences observed suggested that cows of superior genetic fertility potential might have improved reproductive performance, at least in part, because of modestly improved endocrine status, uterine health, and ability to ovulate.

The ovarian function and endocrine phenotypes of lactating dairy cows during the estrous cycle were associated with genomic-enhanced predictions of fertility potential.

The objectives of this prospective cohort study were to characterize associations among genomic merit for fertility with ovarian and endocrine function and the estrous behavior of dairy cows during an entire, non-hormonally manipulated estrous cycle. Lactating Holstein cows entering their first (n = 82) or second (n = 37) lactation had ear-notch tissue samples collected for genotyping using a commercial genomic test. Based on genomic predicted transmitting ability values for daughter pregnancy rate (gDPR) cows were classified into a high (Hi-Fert; gDPR > 0.6 n = 36), medium (Med-Fert; gDPR -1.3 to 0.6 n = 45), and low fertility (Lo-Fert; gDPR < -1.3 n = 38) group. At 33 to 39 DIM, cohorts of cows were enrolled in the Presynch-Ovsynch protocol for synchronization of ovulation and initiation of a new estrous cycle. Thereafter, the ovarian function and endocrine dynamics were monitored daily until the next ovulation by transrectal ultrasonography and concentrations of progesterone (P4), estradiol, and FSH. Estrous behavior was monitored with an ear-attached automated estrus detection system that recorded physical activity and rumination time. Overall, we observed an association between fertility group and the ovarian and hormonal phenotype of dairy cows during the estrous cycle. Cows in the Hi-Fert group had greater circulating concentrations of P4 than cows in the Lo-Fert group from d 4 to 13 after induction of ovulation and from day -3 to -1 before the onset of luteolysis. The frequency of atypical estrous cycles was 3-fold greater for cows in the Lo-Fert than the Hi-Fert group. We also observed other modest associations between genomic merit for fertility with the follicular dynamics and estrous behavior. There were several associations between milk yield and parity with ovarian, endocrine, and estrous behavior phenotypes as cows with greater milk yield and in the second lactation were more likely to have unfavorable phenotypes. These results demonstrate that differences in reproductive performance between cows of different genomic merit for fertility classified based on gDPR may be partially associated with circulating concentrations of P4, the incidence of atypical phenotypes during the estrous cycles, and to a lesser extent the follicular wave dynamics. The observed physiological and endocrine phenotypes might help explain part of the differences in reproductive performance between cows of superior and inferior genomic merit for fertility.

Effect of replacing a portion of inorganic chloride trace minerals with trace mineral amino acid complexes.

The objective was to determine whether replacing a portion of inorganic chloride trace minerals and cobalt carbonate in the diet with AA complexes of trace minerals and cobalt glucoheptonate will improve lactating cow performance, feed efficiency, and calf performance. In a clinical trial, 69 Holstein cows entering second lactation and greater were randomly assigned to 1 of 2 treatments, with the total dietary trace mineral concentration the same between treatments, starting 1 wk after dry off (50 to 57 d before expected parturition) until 154 d in milk (DIM): (1) an inorganic chloride trace mineral (ITM) blend consisting of Zn (75 mg/kg), Mn (65 mg/kg), and Cu (10 mg/kg) as hydroxychlorides and Co (1 mg/kg) as carbonate (n = 37) or (2) partial replacement of ITM with AA complexes of Zn (40 mg/kg), Mn (20 mg/kg), and Cu (3.5 mg/kg) and Co glucoheptonate (1 mg/kg; AATM; Availa-Dairy, Zinpro Corp.; n = 32). Dry matter intake (DMI) was recorded daily from enrollment through wk 8, and milk yields were recorded daily from calving through wk 22. Milk composition and body weights (BW) were collected weekly. Serum samples were analyzed for albumin (Alb), cholesterol (Chol), total bilirubin (Bili), aspartate aminotransferase (AST), haptoglobin, ß-hydroxybutyrate (BHB), and Ca. A liver health index (LHI) was calculated based on Bili, Chol, and Alb concentrations. A liver functionality index (LFI) was calculated to standardize changes in Alb, Chol, and Bili from 4 to 29 DIM. Greater LHI and LFI indicate better health status. Colostrum was analyzed for IgG and Brix, and calf serum was analyzed for IgG. Calf growth was monitored through 9 wk of age (AATM: n = 12, ITM: n = 10). Data were analyzed using SAS software with mixed effects models and repeated-measures analysis, when applicable. Survival analysis for pregnancy by 154 DIM was analyzed by Cox proportional and Kaplan-Meier hazards models. Disorder incidence was tested with Fisher's exact test. Prepartum DMI as a percent of BW was lower in cows fed AATM and not significant postpartum. Cows fed AATM produced more milk from wk 1 to 8 and from wk 1 to 22. Energy-corrected milk yield and colostrum measures did not significantly differ between treatments. A treatment by time interaction was seen for AST and BHB; cows fed AATM tended to have lower AST concentrations at 28 DIM and lower concentrations in BHB through 29 DIM, though not statistically significant. Cows fed AATM had greater LHI at 4 DIM. Haptoglobin, Ca, LFI, hazard of pregnancy, risk to first service, survival curves, or services per pregnancy did not significantly differ. Calf serum IgG and birth weight did not significantly differ between treatments. Calves from dams fed AATM had greater average daily gain than calves from dams fed ITM. Overall, cows fed AATM during the dry period and early lactation had improved postpartum performance and potential health improvements.

Transition cow nutrition and management strategies of dairy herds in the northeastern United States: Part III-Associations of management and dietary factors with analytes, health, milk yield, and reproduction.

The objective was to evaluate relationships between putative periparturient management and dietary factors at the pen and herd levels with metabolic- and inflammation-related analytes, health disorders, milk yield, and reproductive performance. Multiparous and primiparous cows from 72 farms in the northeastern United States were enrolled in a prospective cohort study. Farms were visited 3 times during the prepartum and postpartum periods: during the far-off dry, close-up dry, and fresh periods. Pen measurements were taken at each visit for the pens where cows sampled were housed, and particle size was determined for the total mixed ration for the pen. A survey was used to acquire data on herd-level management variables. Blood samples were collected from the same 11 to 24 cows per farm during the close-up and fresh period visits. Whole blood was analyzed for postpartum ß-hydroxybutyrate (BHB) concentrations, and plasma was analyzed for prepartum and postpartum nonesterified fatty acids (NEFA) and postpartum haptoglobin (Hp) concentrations. Health event, milk yield, and reproductive records were acquired through the records management software program used on the farm. For the pen-level analysis, primiparous and multiparous cows were analyzed separately. For the pen- and herd-level analysis, a simple linear regression was conducted on all possible explanatory variables. Variables were included in the full multivariable general linear model if P < 0.20, and a manual backward stepwise elimination process ensued until all variables had P < 0.10. Our results indicate that pen- and herd-level management factors are associated with blood biomarkers, health, milk yield, and reproductive performance. For the prepartum period, our results support increasing the proportion of particles on the 19-mm sieve of the Penn State Particle Separator, optimizing bunk space, and not overfeeding metabolizable energy (ME), to decrease the prevalence of elevated postpartum NEFA, BHB, and Hp concentrations, decrease disorder incidence, maximize milk yield, and improve pregnancy risk to first service. For the fresh period, our results generally support optimizing bunk space, avoiding commingling, increasing feeding frequency, avoiding high physically effective undigested neutral detergent fiber (NDF) after 240 h of in vitro fermentation and high total fermentable carbohydrate diets while optimizing the inclusion of forage NDF, and ensuring adequate diet ME and metabolizable protein to reduce the prevalence of elevated postpartum NEFA, BHB, and Hp concentrations, minimize disorder incidence, maximize milk yield, and improve pregnancy risk to first service. At the herd level, our results generally support not vaccinating in the calving pen, minimizing the number of prepartum and postpartum pen moves, and avoiding long stays in the calving pen after parturition to reduce the prevalence of elevated biomarker concentrations, decrease disorder incidence, increase milk yield, and improve reproductive performance.

Associations of a liver health index with health, milk yield, and reproductive performance in dairy herds in the northeastern United States.

The objective was to evaluate a liver health index (LHI) by evaluating its association with negative health events, milk yield, and risk of pregnancy within 150 d in milk (DIM). In a retrospective cohort study, an LHI was calculated based on plasma albumin, cholesterol, and bilirubin concentrations for 265 primiparous and 611 multiparous cows 3 to 12 DIM enrolled across 72 farms in the northeastern United States. Mixed effects linear regression models were used to evaluate if (1) metritis (MET), (2) displaced abomasum (DA), (3) clinical ketosis (CK), (4) one or more of the 3 disorders (MET, DA, or CK), (5) 2 or more of the 3 disorders (MET, DA, or CK), or (6) culling within 30 DIM was associated with LHI. Mixed effects linear regression models were used to evaluate if LHI was associated with 305-d mature equivalent milk at the fourth test day (ME305; mean ± standard deviation: 114 ± 13 DIM) and a Cox proportional hazards model was used to evaluate if LHI was associated with pregnancy within 150 DIM. Cows that were diagnosed with MET, DA, CK, one or more of the disorders, 2 or more of the disorders, or were culled within 30 DIM had a lower LHI than cows that were not diagnosed with a disorder or culled. A 1-unit increase in LHI was associated with a 154 ± 38 kg increase in ME305 and a 8% increased risk of pregnancy within 150 DIM [hazard ratio (95% confidence interval): 1.08 (1.03 to 1.14)] for multiparous cows; however, we did not identify a relationship between LHI and ME305 or pregnancy within 150 DIM for primiparous cows. These results suggest that the LHI is associated with health, milk yield, and pregnancy within 150 DIM for multiparous cows and health for primiparous cows; therefore, the LHI can be used as a tool to evaluate transition cow success.

Transition cow nutrition and management strategies of dairy herds in the northeastern United States: Part I-Herd description and performance characteristics.

Our objective was to describe management and herd characteristics of the transition period on freestall dairy herds in the northeastern United States using an on-farm survey and prospective cohort design. Enrolled herds (n = 72) had a median of 900 milking cows (range: 345-2,900) and a rolling herd average of 12,674 kg (standard deviation ± 1,220 kg), and 87.2% (n = 82/94) of fresh pens were milked at least 3×/d. The prevalence of herds with ≥15% of sampled cows with elevated concentrations of nonesterified fatty acids prepartum (≥0.27 mmol/L, 2-14 d before parturition) and postpartum [primiparous: ≥0.60 mmol/L, multiparous: ≥0.70 mmol/L, 3-14 d in milk (DIM)], ß-hydroxybutyrate postpartum (≥1.2 mmol/L, 3-14 DIM), and haptoglobin postpartum (≥1 g/L, 0-12 DIM) was 51%, 51%, 51%, and 57%, respectively. In most herds, cows were moved to a calving pen when showing signs of labor (73.6%; n = 53/72) instead of 0 to 3 d before expected calving (26.4%, n = 19/72). Cows remained in the calving or maternity pen for a median (range) time of 2 (0-24) h after parturition before moving to the next pen. Primiparous cows remained in the first pen moved to after parturition for a longer period than multiparous cows [median (range) days: 12 (1.5-25) vs. 6 (1.5-22)]. Approximately 20% of herds had routine vaccinations administered in the maternity or calving pen, first pen after parturition, or both. Almost all herds (n = 69/72) performed fresh cow health checks; however, only 53% (n = 38/72) locked up all fresh cows daily. More herds housed primiparous and multiparous cows in separate pens during the far-off dry (65.3%; n = 47/72) and high-lactation (81.9%; n = 59/72) periods compared with the close-up dry (31.9%; n = 23/72) and fresh periods (27.8%; n = 20/72). At least half of the pens observed during the far-off dry, close-up dry, and fresh periods had a stocking density <100%. Approximately one-third of pens observed during the far-off dry period had feed pushed up ≤4×/d compared with approximately 15 to 20% of pens observed during the close-up dry, fresh, and high-lactation periods. More than half of the total mixed ration samples acquired from the far-off and close-up dry period visits had greater than the recommended proportion of particles in the 19-mm screen of the Penn State Particle Separator. The results of this observational study illustrated the range of management practices used in freestall herds in this region and lay the groundwork for future hypothesis-driven studies using this sampled population.

Transition cow nutrition and management strategies of dairy herds in the northeastern United States: Part II-Associations of metabolic- and inflammation-related analytes with health, milk yield, and reproduction.

The objectives were as follows: (1) establish cow-level thresholds for prepartum nonesterified fatty acids (NEFA) and postpartum NEFA, ß-hydroxybutyrate (BHB), and haptoglobin (Hp) concentrations associated with negative health events; (2) evaluate cow-level associations between biomarkers and 305-d mature equivalent milk at the fourth test day (ME305) and reproductive performance; and (3) identify herd-alarm levels (proportion of cows sampled above the critical threshold) for biomarkers that are associated with herd-level changes in disorder incidence (displaced abomasum and clinical ketosis), reproductive performance, and ME305. In a prospective cohort study, 1,473 cows from 72 farms were enrolled from the northeastern United States. Blood samples were collected from the same 11 to 24 cows per herd during the late-prepartum and early-postpartum periods. Whole blood was analyzed for postpartum BHB concentrations; plasma was analyzed for prepartum and postpartum NEFA and postpartum Hp concentrations. Critical thresholds for the biomarkers associated with health events for all cows were established using a receiver operating characteristic curve analysis. Poisson, linear mixed effects, and Cox proportional hazards models investigated the association of the biomarkers with health and performance. The prepartum NEFA and Hp threshold associated with culling was ≥0.17 mmol/L and 0.45 g/L, respectively. The postpartum NEFA and BHB thresholds associated with diagnosis of metritis, displaced abomasum, or clinical ketosis were ≥0.46 mmol/L and ≥0.9 mmol/L, respectively. Multiparous cows with prepartum NEFA concentration ≥0.17 mmol/L produced 479 kg less ME305. Multiparous and primiparous cows with postpartum NEFA concentration ≥0.46 mmol/L produced 280 kg less and 446 kg more ME305, respectively. Cows with BHB concentration ≥0.9 and ≥1.1 mmol/L produced 552 kg more ME305 and had a 20% decreased risk of pregnancy within 150 d in milk, respectively; however, multiparous cows with BHB concentration ≥1.5 mmol/L produced 376 kg less ME305. Cows with Hp concentration ≥0.45 g/L produced 492 kg less ME305 and had 28% decreased risk of pregnancy within 150 DIM. Cows with Hp concentration ≥0.45 g/L had 19% decreased pregnancy risk to first service (PRFS). Herds above the herd-alarm levels for prepartum NEFA had a 6.0-percentage unit increase in disorder incidence and a 6.0-percentage unit decrease in 21-d pregnancy rate (PR) for multiparous cows, a 3.9-percentage unit increase in PR and a 5.8-percentage unit increase in the probability of pregnancy for primiparous cows. Herds above the herd-alarm levels for postpartum NEFA had a 5.8- and 4.2-percentage unit increase in disorder incidence for multiparous and primiparous cows, respectively, a 789 kg decrease in ME305 for multiparous cows, and a 6.8- and 6.3-percentage unit decrease and increase in PR and PRFS for multiparous cows, respectively. Herds above the herd-alarm levels for BHB had an 8.5-percentage unit increase in disorder incidence, a 332 and 229 kg increase in ME305 for primiparous and multiparous cows, respectively, and a 3.2-, 5.2-, and 7.0-percentage unit decrease in PR, probability of pregnancy, and PRFS, respectively. Herds above the herd-alarm levels for postpartum Hp had a 5.3-percentage unit increase in disorder incidence. At the cow level and herd level, elevated biomarker concentrations were associated with an increased disorder risk and varied performance responses.

Association of subclinical hypocalcemia dynamics with dry matter intake, milk yield, and blood minerals during the periparturient period.

Subclinical hypocalcemia (SCH) affects many high-producing dairy cows in the postpartum period. Recent work has shown that cows experiencing prolonged or delayed SCH are at increased risk for disease and produce less milk than cows experiencing a transient reduction in or normal concentrations of plasma Ca following parturition. Our objective was to determine the association between different postpartum SCH dynamics with pre- and postpartum dry matter intake (DMI), milk yield, and blood mineral concentrations. Data were retrospectively collected from multiparous Holstein cows (n = 78), and cows were classified into 1 of 4 SCH groups based on mean blood total Ca (tCa) concentrations at 1 and 4 d in milk (DIM): normocalcemic (NC; [tCa] >1.95 mmol/L at 1 DIM and >2.2 mmol/L at 4 DIM, n = 28); transient SCH (tSCH; [tCa] ≤1.95 mmol/L at 1 DIM and >2.2 mmol/L at 4 DIM, n = 27); delayed SCH (dSCH; [tCa] >1.95 mmol/L at 1 DIM and ≤2.2 mmol/L at 4 DIM, n = 6); and persistent SCH (pSCH; [tCa] ≤1.95 mmol at 1 DIM and ≤2.2 mmol/L at 4 DIM, n = 17). Linear mixed models were created to analyze the change in pre- and postpartum DMI, milk yield, and blood mineral concentrations over time as well as differences between SCH groups. Prepartum intake was similar between groups, but the NC and tSCH cows consumed more feed than the pSCH or dSCH cows during the first 3 wk of lactation. The tSCH cows produced more milk than the other 3 groups during the first 6 wk of lactation. Postpartum blood tCa and Mg were different between SCH groups and were highest in the NC cows and lowest in the pSCH cows. Our results suggest that the high level of DMI consumed by the NC and tSCH cows in the postpartum period supported an appropriate homeostatic response to the increased Ca demands of lactation, allowing for higher milk yield compared with their counterparts experiencing delayed or prolonged episodes of SCH.

Effects of feeding synthetic zeolite A during the prepartum period on serum mineral concentration, oxidant status, and performance of multiparous Holstein cows.

The objective of this study was to determine the effects of feeding synthetic zeolite A for 3 wk before expected calving on peripartal serum mineral concentrations, hypocalcemia, oxidant status, and performance. Holstein cows (n = 55) entering their second or greater lactations were assigned randomly to 1 of 2 dietary treatments starting 21 d before expected calving: control (CON: 40% corn silage, 33% wheat straw, and 27% concentrate; n = 29) or experimental [EXP: CON plus zeolite A (X-Zelit, Protekta Inc., Lucknow, ON, Canada/Vilofoss, Graasten, Denmark; n = 26) at an inclusion rate of 3.3% of dry matter, targeting 500 g/d as-fed]. Cows were fed the same postpartum diet and housed in individual tiestalls through 28 d in milk. Cows fed EXP had higher serum Ca concentrations as parturition approached and during the immediate postpartum period. Serum P concentrations were lower for the EXP-fed cows during the prepartum period and the first 2 d of lactation, whereas serum Mg concentrations were lower than those of the CON-fed cows only during the immediate periparturient period. Cows fed EXP had decreased prevalence of subclinical hypocalcemia (SCH) from d -1 through 3 relative to day of parturition, with the largest difference occurring within the first day postpartum. Prepartum dry matter intake tended to be decreased and rumination was decreased in cows fed EXP; however; postpartum dry matter intake, rumination, milk yield, milk component yield, and colostrum measurements did not differ between treatments. Cows fed EXP tended to have increased hazard of pregnancy by 150 d in milk when controlling for parity compared with CON-fed cows; potential reproductive benefits merit further study. This study demonstrated that zeolite A supplementation during the prepartum period results in markedly improved serum Ca concentrations around parturition and similar postpartum performance compared with controls and is effective at decreasing hypocalcemia in multiparous Holstein cows.