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Subfertility represents one major challenge to enhancing dairy production and efficiency. Herein, we use a reproductive index (RI) expressing the predicted probability of pregnancy following artificial insemination (AI) with Illumina 778K genotypes to perform single and multi-locus genome-wide association analyses (GWAA) on 2,448 geographically diverse U.S. Holstein cows and produce genomic heritability estimates. Moreover, we use genomic best linear unbiased prediction (GBLUP) to investigate the potential utility of the RI by performing genomic predictions with cross validation. Notably, genomic heritability estimates for the U.S. Holstein RI were moderate (h2 = 0.1654 ± 0.0317-0.2550 ± 0.0348), while single and multi-locus GWAA revealed overlapping quantitative trait loci (QTL) on BTA6 and BTA29, including the known QTL for the daughter pregnancy rate (DPR) and cow conception rate (CCR). Multi-locus GWAA revealed seven additional QTL, including one on BTA7 (60 Mb) which is adjacent to a known heifer conception rate (HCR) QTL (59 Mb). Positional candidate genes for the detected QTL included male and female fertility loci (i.e. spermatogenesis and oogenesis), meiotic and mitotic regulators, and genes associated with immune response, milk yield, enhanced pregnancy rates, and the reproductive longevity pathway. Based on the proportion of the phenotypic variance explained (PVE), all detected QTL (n = 13; P ≤ 5e - 05) were estimated to have moderate (1.0% < PVE ≤ 2.0%) or small effects (PVE ≤ 1.0%) on the predicted probability of pregnancy. Genomic prediction using GBLUP with cross validation (k = 3) produced mean predictive abilities (0.1692-0.2301) and mean genomic prediction accuracies (0.4119-0.4557) that were similar to bovine health and production traits previously investigated., Competing Interests: Conflicts of interest The authors declare no competing interests., (© The Author(s) 2023. Published by Oxford University Press on behalf of The Genetics Society of America.)

The objective of this study was to characterize incidences of health disorders during early lactation in a large population of Holstein cows calving in 2 seasons across multiple US dairy herds. In addition, cumulative effects of combinations of health-related events on fertility and survival by season of calving and parity number were tested. Data were prospectively collected from a total of 11,729 cows in 16 herds located in 2 regions in the United States [north (7,820 cows in 10 herds) and south (3,909 cows in 6 herds)]. Cows were enrolled at parturition and monitored weekly for disease occurrence, reproductive events, and survival. Health-related events were grouped into reproductive disorders (REP; dystocia, twins, retained fetal membranes, metritis, and clinical endometritis) and other disorders (OTH; subclinical ketosis, mastitis, displaced abomasum, and pneumonia). Counts of health events within 50 d postpartum were added into each of the groups and categorized as 0, 1, 2, 3, and ≥4 for REP and 0, 1, 2, and ≥3 for OTH. Multivariable logistic regression was used for testing potential associations between categories of disease occurrence and outcome variables, including resumption of ovarian cyclicity, pregnancy per artificial insemination (AI), pregnancy loss, and survival up to and after 50 DIM. The incidence of disease varied with season of calving and parity, and these 2 variables were associated with the reproductive and survival outcomes. The size of the detrimental effect of disease incidence on reproduction and survival depended on disease group and varied for each specific outcome. Resumption of ovarian cyclicity decreased as incidences of disorders increased in both REP and OTH categories. Pregnancy at first AI also was smaller in greater number of REP categories, but the effect of number of OTH categories on pregnancy at first AI was not consistent. Similarly, pregnancy loss at first AI was not affected consistently by REP or OTH. Survival was reduced by REP and OTH. The magnitude of these negative effects was variable, depending on season of calving and parity, but consistently increased with the number of health events during early lactation., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

The study is part of a research effort investigating potential associations between genomic variation and fertility of Holstein cows. The objective was to compare the reproductive performance of Holstein cows in 3 categories of 2 reproductive indices (RI) that were developed for the allocation of cows in a ranking for potential fertility, based on the predicted probability of pregnancy. The associations between categories of the developed indices and multiple fertility variables in a large multistate population of Holstein cows were tested. In addition, we analyzed associations among the RI categories with milk yield and survival. Based on phenotypic information from individual cows, 2 reproductive indices (RI1 and RI2) were developed, representing a predicted probability that a cow will become pregnant at first artificial insemination postpartum, as a function of explanatory variables used in a logistic model. Data from a total of 11,733 cows calving in 16 farms located in 4 regions of the United States (Northeast, Midwest, Southeast, and Southwest) were available. Cows were enrolled at parturition and monitored weekly for reproductive events, health status, milk yield, and survival. To develop the indices, potential significant effects were initially tested by univariate analyses. Effects with P ≤ 0.05 were offered to the multivariate analysis, and the final models were determined through backward elimination, considering potentially significant interactions. The final model for RI1 included the random effect of farm and a complement of significant fixed effects as explanatory variables influencing a pregnancy outcome: (1) incidence of retained fetal membranes; (2) metritis; (3) clinical endometritis; (4) lameness at 35 days in milk (DIM); (5) resumption of postpartum ovulation by 50 DIM; (6) season of calving; and (7) parity number. The model for RI2 included (1) parity number; (2) body condition score at 40 DIM; (3) incidence of retained fetal membranes; (4) metritis; (5) resumption of postpartum ovulation by 50 DIM; (6) region; (7) subclinical ketosis; (8) mastitis; (9) clinical endometritis; and (10) milk yield at the first milk test after calving; as well as the interaction effects of postpartum resumption of ovulation by 50 DIM × region; mastitis × region; and milk yield at the first milk test after calving × parity number. Multivariate logistic regression, ANOVA, and survival analysis were used to test the correspondence between the resulting RI and individual fertility, milk yield, and survival from the population. To facilitate the analyses, the resulting RI values were categorized as low for cows in the lowest quartile, medium for cows within the interquartile range, or high for cows in the top quartile. We found consistent agreement between categories of the predicted RI and the measures of fertility and survival collected from individual cows. We conclude that the proposed RI represent a viable approach to refine the allocation of cows into potential low- and high-fertility populations., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Our objectives were to evaluate the efficacy of ampicillin trihydrate for the treatment of metritis in dairy cows compared with ceftiofur hydrochloride and the subsequent effects on pregnancy at first insemination (P/AI). Cows in the first 12 d in milk (DIM) with a uterine discharge score of 5 (watery, reddish or brownish discharge of foul smell) and rectal temperature <39.5°C were diagnosed with metritis based on the fetid discharge, and cows with metritis and rectal temperature ≥39.5°C were diagnosed as having puerperal metritis. Cows with metritis (n=528) were blocked by parity and type of metritis as fetid discharge or puerperal metritis and, within each block, assigned randomly to receive 11mg/kg of ampicillin (n=259) or 2.2mg/kg of ceftiofur (n=269) once daily for 5 d. Day of diagnosis of metritis was considered study d 1. A cohort of 268 cows without metritis was selected randomly at 12 DIM. Rectal temperature was measured in cows with metritis on study d 1 to 7, and 12, and vaginal discharge was scored on study d 5, 7, and 12. Metritis cure was characterized by vaginal discharge score of <5 or by vaginal discharge score of <5 and no fever. At 32±3 DIM, vaginal discharge was scored for diagnosis of purulent vaginal discharge. At 39±3 DIM, endometrial cytology was performed. At 53±3 and 67±3 DIM, ovaries were scanned to determine estrous cyclicity. Pregnancy was evaluated after the first AI. Cure of metritis based on vaginal discharge <5 was greater for ampicillin than ceftiofur on d 5 (37.1 vs. 25.2%) and 7 (57.2 vs. 46.3%), but not on d 12 (82.0 vs. 85.0%). Cure of metritis based on vaginal discharge <5 and no fever was greater for ampicillin than for ceftiofur only on d 7 (50.4 vs. 37.9%), but not on d 5 (23.1 vs. 17.6%) and 12 (66.1 vs. 67.4%). Cows with puerperal metritis had reduced cure compared with cows with fetid discharge on d 5 (30.5 vs. 12.8%), 7 (55.2 vs. 33.6%), and 12 (72.0 vs. 61.1%). The proportion of cows with fever on any day after therapy started did not differ between treatments. Fifty-three percent of cows with metritis based on fetid discharge developed fever after initiating antimicrobial therapy. Cows receiving ampicillin had less prevalence of purulent vaginal discharge than those treated with ceftiofur (57.7 vs. 67.8%), but they were both greater than cows without metritis (21.9%). Prevalence of cytological endometritis did not differ between ampicillin and ceftiofur (30.0 vs. 25.4%), but they were both greater than cows without metritis (14.5%). The proportion of estrous cyclic cows (75.0%) and P/AI did not differ among treatments (ampicillin=28.0% vs. ceftiofur=28.3% vs. without metritis=30.5%). Clinical cure was faster for ampicillin than for ceftiofur, but on study d 12 both treatments resulted in similar cure. Clinical cure was less for cows with puerperal metritis than for cows with fetid uterine discharge. Despite differences in uterine health, P/AI at the first insemination did not differ among treatments., (Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of the current study was to evaluate the effect of GnRH early postpartum on induction of ovulation, uterine health, and fertility in dairy cows. Holstein cows without a corpus luteum (CL) at 17 ± 3 DIM were assigned randomly to receive i.m. GnRH (n = 245) at 17 ± 3 and 20 ± 3 DIM or remain as controls (n = 245). Ovaries were scanned by ultrasonography twice weekly totaling 4 examinations. Ovulation was characterized by the appearance of a CL ≥ 20 mm at any ultrasound or CL <20mm in 2 consecutive examinations. Clinical and cytological endometritis were diagnosed at 35 DIM. Compared with control, GnRH increased ovulation up to 3.5d after the last treatment (78.7 vs. 45.0%) and did not affect the prevalence of clinical endometritis (23.9 vs. 18.6%) or cytological endometritis (30.9 vs. 32.8%). Prevalence of clinical endometritis increased in cows that had calving problems (32.6 vs. 15.9%) and metritis (40.6 vs. 15.8%). Metritis increased prevalence of cytological endometritis (50.7 vs. 23.5%). Treatment with GnRH did not affect pregnancy per artificial insemination at 32 (37.6 vs. 38.6%) or 74 d after artificial insemination (35.0 vs. 31.5%), but reduced pregnancy loss (6.8 vs. 18.1%). No overall effect of GnRH treatment on hazard of pregnancy was observed; however, an interaction between GnRH treatment and ovulation showed that GnRH-treated cows that ovulated had increased hazard of pregnancy by 300 DIM compared with GnRH-treated and control cows that did not ovulate (hazard ratio=2.0 and 1.3, respectively), but similar to control cows that ovulated (hazard ratio=1.1). Gonadotropin-releasing hormone early postpartum induced ovulation without affecting uterine health, but failed to improve pregnancy per artificial insemination or time to pregnancy, although it reduced pregnancy loss., (Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Our objectives were to determine the effects of early ovulation on fertility and uterine health of dairy cows. Four hundred and forty-five Holstein cows (185 primiparous and 260 multiparous) from five herds were used. Blood samples were collected at 21, 35 and 49 days in milk (DIM) and cows were considered to be cyclic at 21 DIM (Cyc21) if serum progesterone (P4) concentration was above 1 ng/ml, cyclic by 49 DIM (Cyc49) if P4 concentration was above 1 ng/ml at 35 or 49 DIM, or not cyclic (NotCyc) if P4 concentration was below 1 ng/ml at all sample times. Endometrial cytology for diagnosis of subclinical endometritis was examined at 49 DIM in a subset of 414 cows. Cows in the group Cyc21 had increased hazard of insemination, for the first service, compared with cows in Cyc49 [hazard ratio (HR) = 1.40; 95% CI = 1.10-1.79; p = 0.006] and NotCyc (HR = 2.07; 95% CI = 1.52-2.82; p < 0.001). Cows in the Cyc49 group also had increased hazard of insemination compared with cows in the NotCyc group (HR = 1.47; 95% CI = 1.13-1.93; p = 0.005). Median days to insemination were, respectively, 71, 76 and 96 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in Cyc21 had greater first service pregnancy per AI than Cyc49 [38.6 vs 28.1%; adjusted odds ratio (AOR) = 1.67; 95% CI = 1.01-2.75; p = 0.04] and NotCyc (38.6 vs 23.6%; AOR = 2.08; 95% CI = 1.08-4.00; p = 0.03). Pregnancy per AI was similar in Cyc49 and NotCyc cows (28.1 vs 23.6%; AOR = 1.25; 95% CI = 0.70-2.24; p = 0.45). Cows in Cyc21 had increased hazard of pregnancy up to 300 DIM compared with Cyc49 (HR = 1.52; 95% CI = 1.17-1.96; p = 0.002) and NotCyc (HR = 1.98; 95% CI = 1.41-2.78; p < 0.001). Cows in Cyc49 tended to have increased hazard of pregnancy compared with NotCyc (HR = 1.31; 95% CI = 0.96-1.77; p = 0.09). Median days to pregnancy were, respectively, 103, 147 and 173 for cows in Cyc21, Cyc49 and NotCyc groups. Cows in the Cyc21 group had decreased prevalence of subclinical endometritis compared with cows in the NotCyc group (29.9 vs 43.7%; AOR = 0.53; 95% CI = 0.29-0.97; p = 0.04); however, the prevalence did not differ from the Cyc49 group (29.9 vs 39.1%; AOR = 0.68; 95% CI = 0.41-1.14; p = 0.15). Cyc49 cows had similar prevalence of subclinical endometritis compared with NotCyc cows (AOR = 0.77; 95% CI = 0.46-1.29; p = 0.32). Early postpartum ovulation was associated with improved uterine health and fertility., (© 2009 Blackwell Verlag GmbH.)

Objectives were to evaluate effects of feeding a calcium salt rich in linoleic and trans-octadecenoic acids (LTFA) on synthesis of prostaglandin F(2alpha) based on its metabolite (PGFM), uterine involution and pregnancy rates in lactating dairy cows. Five hundred and eleven Holstein cows were blocked according to parity, body condition score and milk yield in the previous lactation. Primiparous and multiparous cows were randomly assigned to one of the two treatments consisting of calcium salt (2% diet dry matter) of either palm oil (PO) or LTFA from 25 days prepartum to 80 days of lactation. Cows were time-inseminated at 70 +/- 3 days postpartum. Feeding LTFA tended (p = 0.08) to decrease the incidence of puerperal metritis (15.1% vs 8.8%). Primiparous cows supplemented with LTFA showed larger increase in plasma PGFM concentration at day 1 postpartum (17018 vs 6897 pm). Pregnancy rate after first insemination tended (p = 0.07) to be greater at 27 days after insemination (37.9% vs 28.6%), and was greater (p = 0.05) at 41 days after insemination (35.5% vs 25.8%) for cows fed LTFA compared with PO. These results indicate that unsaturated fatty acids fed in a rumen inert form have the potential to modulate reproductive events and improve pregnancy rates in lactating dairy cows.

The objectives were to determine the effects of PGF(2alpha) treatment on the prevalence of subclinical endometritis (SCE) and fertility of dairy cows. A total of 406 Holstein cows (167 primiparous and 239 multiparous) from 5 herds were used. Uterine lavage for diagnosis of SCE, PGF(2alpha) treatment, evaluation of body condition scores (BCS), and collection of blood samples for estrous cyclicity determination were performed at 21, 35, and 49 d in milk (DIM). Polymorphonuclear cells (PMN) were quantified and thresholds for diagnosing SCE were selected by receiver operating characteristics analysis. Cows classified as having SCE at 35 DIM (>or=6.5% PMN) and 49 DIM (>or=4.0% PMN) had increased time to pregnancy; however, cows classified as having SCE at 21 DIM (>or=8.5% PMN) did not. Median days to pregnancy were delayed by 30 (151 vs. 121 d) and 40 (169 vs. 129) d for cows classified as having SCE at 35 and 49 DIM, respectively. Treatment with PGF(2alpha) did not affect the prevalence of SCE either at 35 (37.9 vs. 38.4%) or at 49 DIM (34.0 vs. 40.4%). Treatment with PGF(2alpha) did not affect time to first insemination (AI; median 76 DIM for cows treated with PGF(2alpha); 79 DIM for control. Nonetheless, PGF(2alpha) treatment increased pregnancy to first AI in all the cows (35.5 vs. 24.1%) and hazard ratio (HR) of pregnancy in cows with BCS

Objectives were to determine the effects of intrauterine (i.u.) infusion of ceftiofur hydrochloride on uterine health and fertility of dairy cows already receiving PGF(2alpha) for estrous synchronization. Holstein cows at 44 +/- 3 d in milk (DIM) were blocked by parity and diagnosis of metritis in the first 14 DIM and of other illnesses and, within each block, randomly assigned to 1 of 2 treatments: a single i.u. infusion of 125 mg of ceftiofur hydrochloride at 44 +/- 3 DIM (ceftiofur, n = 396), or no i.u. infusion (control, n = 416). All cows received 25 mg of PGF(2alpha) at 37 +/- 3 and 51 +/- 3 DIM as part of an estrous synchronization protocol. A subset of 547 cows was evaluated for clinical endometritis immediately before treatment, and 202 cows had an aseptic uterine sample collected before the injection of PGF(2alpha) at 51 +/- 3 DIM for bacteriology and diagnosis of subclinical endometritis (> or =5% neutrophils). Pregnancy on d 38 +/- 3 and 180 +/- 7 after the first artificial insemination, pregnancy loss, and interval from calving to pregnancy in the first 300 DIM were evaluated. The proportions of cows diagnosed with clinical endometritis before treatment were similar between ceftiofur and control treatments. Intrauterine infusion with ceftiofur did not influence prevalence of subclinical endometritis and positive uterine culture 7 d after treatment; however, it reduced the prevalence of positive uterine culture in cows with clinical endometritis (29.0 vs. 51.4%) and reduced the overall prevalence of Arcanobacterium pyogenes (1.0 vs. 7.6%) at 51 +/- 3 DIM. Cows with clinical endometritis had increased prevalence of A. pyogenes (10.3 vs. 1.5%), Escherichia coli (5.9 vs. 0.75%), and overall positive uterine culture (41.2 vs. 22.4%); however, cows with subclinical endometritis only had an increased prevalence of A. pyogenes (10.2 vs. 1.5%). Ceftiofur did not affect pregnancy per artificial insemination in all cows or in cows previously diagnosed with metritis or clinical endometritis. Interval to pregnancy was similar for control and ceftiofur cows. Intrauterine infusion of ceftiofur hydrochloride reduced the prevalence of uterine infection in cows with clinical endometritis, and the prevalence of A. pyogenes, but did not affect the prevalence of subclinical endometritis or fertility of dairy cows already receiving PGF(2alpha).

The objective was to determine if reducing the interval from presynchronization to the first GnRH injection (G1) of a timed artificial insemination (AI) protocol improves pregnancy per AI. One thousand two hundred fourteen Holstein cows, at 37 +/- 3 d in milk (DIM), were stratified by parity, DIM, and milk yield in the first month postpartum and randomly assigned to control (n = 412), 2 injections of PGF2alpha at 37 +/- 3 and 51 +/- 3 DIM, then enrolled in a timed AI protocol 14 d later; PShort (n = 410), 2 injections of PGF2alpha at 40 +/- 3 and 54 +/- 3 DIM, then enrolled in a timed AI protocol 11 d later; or PShortG (n = 392), same as PShort, but with an injection of GnRH 7 d before G1. All cows received the same timed AI protocol (d 65, G1; d 72, PGF2alpha; d 73, 1 mg of estradiol cypionate; d 75, AI). A subset of 1,000 cows had their ovaries examined by ultrasonography at G1 and 7 d later when PGF2alpha of the timed AI was given to determine presence of corpus luteum (CL) and ovulation to G1. Pregnancy was diagnosed on d 38 after timed AI, and pregnant cows were reevaluated for pregnancy 4 wk later. Altering the interval between presynchronization and G1 did not affect the proportion of cows with a CL at G1, but GnRH 7 d before G1 increased the proportion of cows with a CL. Ovulation to G1 was greater for 11 compared with the 14 d interval, but GnRH did not improve ovulation. The increased ovulation to G1 when the interval was reduced from 14 to 11 d was observed only in cows with a CL at G1, but treatment did not affect ovulation in cows without a CL at G1. Treatment affected the pregnancy per AI on d 38 and 66 after insemination, and they were greater for the 11 compared with 14-d interval, but addition of GnRH did not improve pregnancy per AI. Cows ovulating to G1 had greater pregnancy per AI regardless of whether or not they had a CL at G1. Reducing the interval from presynchronization to initiation of the timed AI protocol from 14 to 11 d increased ovulation to G1 and pregnancy per AI in lactating dairy cows.