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Anogenital distance (AGD), defined as the distance from the center of the anus to the base of the clitoris, in lactating dairy cows of first and second parity, has been reported to be inversely related to fertility and moderately heritable. Thus, AGD may be a useful reproductive phenotype for future genetic selection to improve fertility. The objectives of this study were to (1) characterize AGD in nulliparous dairy heifers; and (2) determine if the inverse relationship between AGD and fertility, found in lactating dairy cows, is also evident in nulliparous heifers. We measured AGD in 1,692 Holstein heifers from 16 herds in Western Canada (Alberta and British Columbia) and one herd in the United States (Washington State). Data were analyzed using MEANS, UNIVARIATE, LOGISTIC, ROC, GLIMMIX, and LIFETEST procedures of SAS (SAS Institute Inc.). Mean (±standard deviation) age at AGD measurement was 13.9 ± 1.5 mo, and AGD was normally distributed with a mean of 107.3 ± 10.5 mm, ranging from 69 to 142 mm. With every 1-mm increase in AGD, the predicted probability of pregnancy was reduced by 1.9%. Receiver operating characteristic curve analysis was used to determine the optimum threshold AGD that predicted the probability of pregnancy. Based on the optimum threshold AGD, data from heifers were categorized into short (≤110 mm) and long (>110 mm) AGD groups, and associations between AGD groups and fertility measures were determined. Heifers with short AGD required fewer services per conception (1.5 vs. 1.7) than heifers with long AGD. Consequently, heifers with short AGD conceived earlier (448.4 vs. 454.3 d) and had greater pregnancy to first AI than those with long AGD (58.3 vs. 49.6%). Moreover, heifers with long AGD had reduced hazard (hazard ratio of 0.59) for pregnancy up to 450 d of life compared with those with short AGD. In summary, AGD was normally distributed and highly variable in the population. In addition, an inverse relationship between AGD and fertility measures in nulliparous heifers was evident, confirming an earlier report of a similar relationship in lactating dairy cows. These findings strengthen the potential for AGD to be used as a fertility trait and management tool in future selection programs., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objectives of this study were to (1) characterize the distribution and variability of plasma anti-Müllerian hormone (AMH) concentration; (2) evaluate factors associated with phenotypic variation in plasma AMH; (3) examine the associations between categories of plasma AMH and reproductive outcomes [pregnancy to first artificial insemination (P/AI), and pregnancy rates within 21, 42, and 84 d after the mating start date (MSD)]; (4) estimate pedigree and genomic heritability for plasma AMH; and (5) identify and validate SNP associated with phenotypic variation in plasma AMH. Plasma AMH concentration (pg/mL) was determined from a blood sample collected (mean ± standard deviation) 10 ± 2 d after first insemination at detected estrus (IDE) in 2,628 first- and second-parity Irish dairy cows. Overall, plasma AMH had a positively skewed distribution with mean (± standard deviation), median, minimum, and maximum concentrations of 326 ± 231, 268, 15, and 2,863 pg/mL, respectively. Plasma AMH was greatest for Jersey, followed by Holstein × Jersey, Holstein × Norwegian Red, and Holstein cows (410, 332, 284, and 257 pg/mL, respectively). Second-parity cows had greater plasma AMH than first-parity cows (333 vs. 301 pg/mL, respectively). Samples collected at 7 and 8 d after first IDE had lesser plasma AMH than those collected on d 9, 10, 11, 12, and 13 after first IDE (291 and 297 vs. 317, 319, 331, 337, and 320 pg/mL). Plasma AMH was not associated with either body condition score at first IDE or the interval from calving to MSD. Cows were categorized into low (≤150 pg/mL; n = 526; lowest 20%), intermediate (>150 to ≤461 pg/mL; n = 1,576; intermediate 60%), and high AMH (>461 pg/mL; n = 526; highest 20%) groups based on plasma AMH, and associations with reproductive outcomes were tested. Cows with high and intermediate plasma AMH had 1.42- and 1.51-times-greater odds of becoming pregnant within 84 d after the MSD than those with low plasma AMH (90.3 and 90.8 vs. 86.8%, respectively); however, P/AI and pregnancy rate within 21 and 42 d after the MSD did not differ among AMH categories. Plasma AMH was moderately heritable (pedigree heritability of 0.40 ± 0.06 and genomic heritability of 0.45 ± 0.05), and 68 SNP across Bos taurus autosomes 7 and 11 were associated with phenotypic variation in plasma AMH. Out of 68 SNP, 42 were located in a single quantitative trait locus on Bos taurus autosome 11 that harbored 6 previously identified candidate genes (NR5A1, HSPA5, CRB2, DENND1A, NDUFA8, and PTGS) linked to fertility-related phenotypes in dairy cows., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The evaluation of anogenital distance (AGD), the distance from the center of the anus to base of the clitoris, as a potential fertility trait for genetic selection in dairy cows has generated recent interest. The objectives of this cross-sectional observational study were to (1) characterize the distribution and variability of AGD, (2) determine factors associated with AGD, (3) estimate heritability for AGD, (4) identify single nucleotide polymorphisms (SNP) associated with phenotypic variation of AGD, and (5) validate the relationship between categories of AGD and fertility in Irish Holstein-Friesian cows. Anogenital distance was measured using digital calipers in 1,180 Holstein cows (mean ± standard deviation: 225 ± 79 d in milk) from 10 dairy herds located in Munster, Ireland. In addition, age (yr), weight (kg), height at hip (cm), and body condition score (BCS) at the time of AGD measurement were determined in a subset of 281 cows. Genotype information available from 908 cows was subsequently imputed to the Illumina Bovine High Density BeadChip (Illumina Inc., San Diego, CA) for genome-wide association analysis of phenotypic variation in AGD. Overall, AGD had a normal distribution and high variability (mean ± standard deviation; 119.2 ± 11.6 mm). Anogenital distance was weakly but positively associated with cow age, hip height, and body weight, and negatively associated with BCS; the phenotypic variation in AGD that was explainable by these variables was small (coefficient of determination; R 2 = 0.09, 0.06, 0.10, and 0.02, respectively). The estimated heritability for AGD was 0.37 (standard error of mean ± 0.08). Six SNP of suggestive significance were identified on Bos taurus autosomes 6, 15, 20, and 26; however, none of these SNP was related to previously identified candidate genes for fertility. Cows were categorized into quartiles (Q1; 86 to 111 mm; n = 311, Q2; 112 to 120 mm; n = 330; Q3; 121 to 127 mm; n = 265, and Q4; 128 to 160 mm; n = 274) based on AGD and the association with reproductive outcomes examined (21-d submission rate, pregnancy to first AI, pregnancy rate within 21, 42 and 84-d after the farm mating start date, and number of times bred). None of the reproductive variables differed significantly between AGD categories. In summary, despite identification of high variability and moderate heritability for AGD in Irish Holstein-Friesian cows, reproductive outcomes did not differ between categories of AGD. This latter result differs from our previous finding of an inverse relationship between AGD and pregnancy outcomes in first- and second-parity Canadian Holstein cows, emphasizing the need to test and validate this new phenotype in diverse cow populations., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Our objectives were to characterize parameters of luteal activity based on milk progesterone concentration (P4c) data from before and after artificial insemination (AI) and to evaluate their potential association with fertility in Holstein cows. Records of AI events (n = 4,353) and of milk P4c (n = 158,961) obtained through an in-line milk analysis system (Herd Navigator, DeLaval International, Tumba, Sweden) from 1,891 lactations of 1,423 Holstein cows were evaluated. Milk P4c (ng/mL) were measured every 2.2 ± 1.9 d (mean ± standard deviation) between 23.6 ± 7.3 and 185.3 ± 56.7 d in milk. Variations in milk P4c of consecutive records were used to determine onset of luteal phase (increase in P4c from <5.0 to ≥5.0 ng/mL), luteal phase length (period, in days, of P4c ≥5.0 ng/mL), cessation of luteal phase (decline from ≥5.0 to <5.0 ng/mL, designated as P4c-decline), and pregnancy (AI followed by a luteal phase that remained uninterrupted until 50 d post-AI). The length of the luteal phase preceding AI, the highest P4c (P4c peak) during the luteal phase preceding AI, the lowest P4c preceding AI (P4c pre-AI) that followed a P4c-decline, and the interval between P4c-decline and AI were evaluated, as well as the interval between AI and onset of luteal phase, and P4c at early diestrus (4.5 ± 0.6 d post-AI), mid diestrus (10.0 ± 0.6 d post-AI), and late diestrus (14.1 ± 0.6 d post-AI). Data were analyzed using logistic regressions, and comparisons made based on quartiles and cut-points established by receiver operating characteristic curve analysis. Overall probability of pregnancy was 32.0%. Parameters associated with reduced probability of pregnancy (represented as percentage points decrease in the probability of pregnancy) were (1) luteal phase length >14.4 d (7.6% decrease), (2) P4c peak ≤24.7 ng/mL (4.5% decrease), (3) P4c pre-AI >0.5 ng/mL (5.5% decrease), (4) interval between P4c-decline and AI of >1.6 d (4.0% decrease), (5) interval between AI and onset of luteal phase of <7 or >11 d (9.3 and 12.1% decrease, respectively), and (6) P4c at early diestrus ≤0.7 or >3.5 ng/mL (15.2 and 6.7% decrease, respectively), (7) P4c at mid diestrus ≤12.4 ng/mL (12.5% decrease), and (8) P4c at late diestrus ≤22.7 ng/mL (9.7% decrease). The parameters of luteal activity associated with reduced probability of pregnancy established here could be used as benchmarks while developing recommendations to improve reproductive performance in herds using in-line milk progesterone monitoring., (The Authors. Published by FASS Inc. and Elsevier 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 objectives of this study were to determine (1) factors associated with serum concentration of insulin-like growth factor-1 (IGF-1); (2) the relationship between serum IGF-1 concentration during the first week postpartum and ovarian cyclicity status by 35 d postpartum (DPP); (3) an optimum serum IGF-1 concentration threshold predictive of pregnancy to first artificial insemination (P/AI), including its diagnostic values; (4) the associations among categories of serum IGF-1 concentration and reproductive outcomes (P/AI and pregnancy risk up to 150 and 250 DPP); and (5) single nucleotide polymorphisms (SNP) associated with phenotypic variation in serum IGF-1 concentration in dairy cows. Serum IGF-1 concentration was determined at 7 (±2.4; ±standard error of the mean) DPP in 647 lactating Holstein cows (213 primiparous, 434 multiparous) from 7 herds in Alberta, Canada. The overall mean, median, minimum, and maximum serum IGF-1 concentrations during the first week postpartum were 37.8 (±1.23), 31.0, 20.0, and 225.0 ng/mL, respectively. Herd, age, parity, precalving body condition score, and season of blood sampling were all identified as factors associated with serum IGF-1 concentrations. Although serum IGF-1 concentration during the first week postpartum had no association with ovarian cyclicity status by 35 DPP in primiparous cows, it was greater in cyclic than in acyclic multiparous cows (32.2 vs. 27.4 ng/mL, respectively). The optimum serum IGF-1 thresholds predictive of P/AI were 85.0 ng/mL (sensitivity = 31.9%; specificity = 89.1%) and 31.0 ng/mL (sensitivity = 45.5%; specificity = 66.9%) for primiparous and multiparous cows, respectively. When cows were grouped into either high or low IGF-1 categories (greater or less than or equal to 85.0 ng/mL for primiparous cows and greater or less than or equal to 31.0 ng/mL for multiparous cows, respectively), primiparous cows with high IGF-1 had 4.43 times greater odds of P/AI and a tendency for higher pregnancy risk up to 150 DPP than those with low IGF-1, but not up to 250 DPP. Likewise, multiparous cows with high IGF-1 had 1.61 times greater odds of P/AI than those with low IGF-1. Pregnancy risk up to 150 and 250 DPP, however, did not differ between IGF-1 categories in multiparous cows. Moreover, 37 SNP across 10 Bos taurus autosomes were associated with variation in serum IGF-1 concentration, and 4 previously identified candidate genes related to fertility that were in linkage disequilibrium with some of these SNP were also identified., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objectives of this study were to (1) evaluate factors associated with variation in circulating anti-Müllerian hormone (AMH) concentrations, (2) establish an optimum AMH threshold predictive of pregnancy to first artificial insemination (P/AI), (3) examine the relationship between AMH and fertility (P/AI, pregnancy loss between 30 and 60 d after artificial insemination, and pregnancy risk up to 250 d postpartum), and (4) identify quantitative trait loci associated with phenotypic variation of AMH concentrations in dairy cows. Serum AMH concentrations (pg/mL) were determined at 7 ± 2.4 d postpartum in 647 lactating Holstein cows (213 primiparous, 434 multiparous) from 1 research and 6 commercial dairy herds in Alberta, Canada. Of these, 589 cows were genotyped on the 26K Bovine BeadChip (Neogen Inc., Lincoln, NE) and subsequently imputed to the Illumina Bovine High Density BeadChip (Illumina, San Diego, CA) for genome-wide association analysis for variation in serum AMH concentrations. Factors associated with variation in serum AMH concentrations and the relationship between categories of AMH and aforementioned fertility outcomes were evaluated only in a subset of 460 cows that had a complete data set available. The overall mean (±standard error of the mean), median, minimum, and maximum AMH concentrations were 191.1 ± 6.3, 151.7, 13.9, and 1,879.0 pg/mL, respectively. The AMH concentrations were not associated with herd, precalving body condition score, postpartum week, and season of sampling; the lactation number, however, had a quadratic relationship with serum AMH concentrations (116.2, 204.9 204.5, and 157.9 pg/mL for first, second, third, and ≥fourth lactation, respectively). The optimum AMH threshold predictive of P/AI could not be established because the receiver operating characteristic curve analysis model was nonsignificant. Categories of AMH [low (<83.0 pg/mL; n = 92), intermediate (≥83.0 to ≤285.0 pg/mL; n = 276), and high (>285.0 pg/mL; n = 92) based on lowest 20%, intermediate 60%, and highest 20% serum AMH) had no associations with P/AI (34, 43, and 40%), pregnancy loss between 30 and 60 d after artificial insemination (20, 12, and 8%), or pregnancy risk up to 250 d postpartum. One candidate gene associated with AMH production [AMH gene on Bos taurus autosome (BTA) 7] and 4 candidate genes related to embryo development (SCAI and PPP6C genes on BTA11 and FGF18 and EEF2K genes on BTA20 and BTA25, respectively) were in linkage disequilibrium with single nucleotide polymorphisms associated with phenotypic variation in serum AMH in dairy cows., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Anogenital distance (AGD) serves as a marker for prenatal androgenization, reproductive development, and fertility in humans and rodents. The primary objectives of this observational study in lactating dairy cows were to (1) characterize the distribution and variability of AGD, (2) determine the relationship among AGD and potential postnatal AGD determinants of age and height, and (3) evaluate the associations between AGD and pregnancy to first artificial insemination (P/AI) and cumulative pregnancy by 250 d in milk (DIM) within parity groups (first, second, and third+ parities). The secondary objective was to evaluate the association between AGD and testosterone concentrations. The AGD (mm), age (yr), and height at hip (cm) at the time of AGD determination, and aforesaid reproductive outcomes were determined in 921 Holstein cows (first, second, and third+ parity; n = 360, 256, and 305, respectively). Plasma concentrations of testosterone were determined in a subset of 93 cows. Overall, AGD had a normal distribution and high variability [mean (±standard deviation); 131.0 ± 12.2 mm], was weakly associated with cow age and height (coefficient of determination = 0.09 and 0.04, respectively), and had an inverse relationship with P/AI in first- and second-parity cows, but not in third+ parity cows. For every 1 mm increase in AGD, the odds of P/AI decreased by 3.4 and 2.4% for first- and second-parity cows, respectively. The optimal AGD threshold to predict probability of P/AI was 127.1 mm for both first- (sensitivity: 66.4; specificity: 56.6%) and second-parity cows (sensitivity: 46.0; specificity: 70.4%). Accordingly, first- and second-parity cows were categorized into either short or long AGD (≤ or >127.1 mm), and associations with reproductive outcomes were evaluated. First-parity cows with long AGD had lower P/AI (30.9 vs. 53.6%) and decreased likelihood (hazard ratio: 0.68) of pregnancy by 250 DIM than those with short AGD. Similarly, second-parity cows with long AGD had reduced P/AI (28.3 vs. 44.4%) and a tendency for decreased likelihood (hazard ratio: 0.76) of pregnancy by 250 DIM than in cows with short AGD. The association between AGD and testosterone was weak and nonsignificant. In summary, AGD in Holstein cows was normally distributed, highly variable, and weakly associated with age and height. Besides, AGD had an inverse relationship with P/AI and cumulative pregnancy by 250 DIM in first- and second-parity cows; however, such a relationship was not evident in older (third+ parity) cows., (The Authors. Published by the Federation of Animal Science Societies and Elsevier 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/3.0/).)

The primary objective was to determine the variability and repeatability of GnRH-induced LH responses. The secondary objective was to evaluate the associations among plasma LH, FSH, estradiol (E2), and progesterone (P4) concentrations. One hundred lactating Holstein cows (35 primiparous, 65 multiparous) were initially subjected to a presynchronization protocol (d 0, PGF 2α ; d 3, GnRH) followed 7 d later by Ovsynch (d 10, GnRH; d 17, PGF 2α ; 56 h later, GnRH) and timed artificial insemination 16 h after the last GnRH. Blood samples were collected immediately before the GnRH injection of presynchronization and the second GnRH of Ovsynch to determine plasma concentrations of LH, FSH, and P4. A second blood sample was collected 2 h after each of the above GnRH injections to determine GnRH-induced LH and FSH concentrations. Plasma concentrations of E2 were also determined in samples collected immediately before the second GnRH of Ovsynch. Cows that (1) had higher LH concentrations at 0 h than at 2 h after GnRH, (2) showed an ongoing spontaneous LH surge, (3) did not respond to GnRH, and (4) had P4 ≥ 0.5 ng/mL at GnRH of presynchronization and the second GnRH of Ovsynch were excluded from the analysis. The variability (coefficient of variation) and repeatability [between animal variance/(within animal variance + between animal variance)] of GnRH-induced LH response were determined from samples collected 2 h after the GnRH of presynchronization and the second GnRH of Ovsynch. The associations among plasma LH, FSH, E2, and P4 were determined at the second GnRH of Ovsynch. Mean (±SEM) LH concentrations before GnRH were 0.5 ± 0.04 and 0.6 ± 0.03 ng/mL, whereas mean LH concentrations 2 h after GnRH were 9.8 ± 1.0 and 12.1 ± 0.8 ng/mL at GnRH of presynchronization and the second GnRH of Ovsynch, respectively. The variability of GnRH-induced LH was 76.1 and 52.1% at GnRH of presynchronization and the second GnRH of Ovsynch, respectively. The repeatability estimate for GnRH-induced LH concentration between GnRH of presynchronization and Ovsynch assessments was 0.10. Plasma concentrations of LH were positively associated with FSH and E2 (r = 0.61 and 0.30, respectively) and negatively associated with P4 (r = -0.46) at the second GnRH of Ovsynch. In summary, GnRH-induced LH responses were highly variable and unrepeatable, and LH concentrations were positively associated with FSH and E2 and negatively associated with P4., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Pregnancy per AI (P/AI) following the use of 1 of 2 timed AI (TAI) protocols and 2 different intervals between TAI and resynchronization were compared in heifers that were inseminated with either conventional or sex-sorted semen. Holstein heifers (n = 317; 527 inseminations) were submitted to a 5-d Cosynch protocol with (+) or without (-) GnRH at the time of controlled internal drug release (CIDR) insertion on d 0, CIDR removal and a single PGF 2α treatment on d 5, and TAI plus GnRH on d 8 (72 h later). Visual estrus detection (ED) was conducted on d 6 in the afternoon and d 7 in the morning and heifers observed in estrus were artificially inseminated on d 7 in the afternoon. Heifers were alternately assigned conventional or sex-sorted semen. Pregnancy was diagnosed by ultrasound 27 and 42 d after AI, and heifers diagnosed as nonpregnant were resynchronized, up to 3 times, starting on d 27 or 34 to provide an interbreeding interval of 35 or 42 d. Overall, TAI protocol had no effect on P/AI at 27 or 42 d after artificial insemination or on pregnancy loss, but P/AI following the first service tended to be higher in the -GnRH TAI group (66.3 vs. 56.8%). Pregnancy per AI at 27 d (61.9 vs. 55.5%) tended to differ between conventional and sex-sorted semen. Heifers artificially inseminated based on ED tended to have a greater P/AI (67.6 vs. 58.2%) and had decreased pregnancy loss (0.0 vs. 4.1%) than those submitted to TAI. A greater number of heifers in the -GnRH TAI protocol were artificially inseminated on ED than the +GnRH TAI protocol (21.5 vs. 13.7%). No difference in P/AI was observed between the 35- and 42-d interbreeding intervals; however, more heifers in the 42-d group were artificially inseminated based on ED than in the 35-d group (22.7 vs. 7.8%). A 5-d Cosynch+CIDR TAI protocol without the initial GnRH and with a single PGF 2α at CIDR removal is an acceptable alternative to achieve high P/AI when either conventional or sex-sorted semen is used in Holstein heifers. Breeding heifers based on detected estrus increases labor, but has the potential to increase fertility., (Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objectives were to determine the effects of supplemental fat (no oilseed vs. oilseed) during late gestation and the source of fat (canola vs. sunflower seed), on dry matter intake (DMI), plasma metabolite concentrations, milk production and composition, calf birth weight, postpartum health disorders, ovarian function and reproductive performance in dairy cows. Pregnant Holstein cows, blocked by body condition and parity, were assigned to 1 of 3 diets containing rolled canola seed (high in oleic acid; n=43) or sunflower (high in linoleic acid; n=45) at 8% of dry matter, or no oilseed (control; n=43), for the last 35±2 d of pregnancy. After calving, all cows received a common lactation diet. Blood samples were collected at wk -3 (i.e., 2 wk after initiation of prepartum diets) and at wk +1, +2, +3, +4 and +5 postpartum to determine the concentration of fatty acids (mEq/dL), β-hydroxybutyrate (mg/dL), and glucose (mg/dL). Ovarian ultrasonography was performed twice weekly to determine the first appearance of dominant (10mm) and preovulatory-size (≥16mm) follicles, and ovulation. Uterine inflammatory status based on the proportion of polymorphonuclear leukocytes (PMN; subclinical endometritis: >8% PMN) was assessed at d 25±1 postpartum. Significant parity by treatment interactions were observed for DMI and milk yield. Prepartum oilseed supplementation, more specifically sunflower seed supplementation, increased postpartum DMI in primiparous cows without affecting prepartum DMI or milk yield. Contrarily, in multiparous cows, prepartum oilseed supplementation decreased both prepartum and postpartum DMI and milk yield during the first 2 wk. Regardless of parity, prepartum feeding of canola reduced postpartum DMI compared with those fed sunflower. Mean fatty acids concentrations at wk -3 were greater in cows given supplemental oilseed than those fed no oilseeds. Gestation length and calf birth weight were increased in cows given supplemental oilseed prepartum compared with cows fed no oilseeds, and a disproportionate increase in the birth weight of female calves was evident in cows fed oilseed. Total reproductive disorders tended to be greater in cows fed supplemental oilseed than those fed no oilseed (42 vs. 23%). Furthermore, cows fed sunflower seed had greater incidences of dystocia (35 vs. 18%) and total health disorders (52 vs. 32%) than those fed canola seed. Added oilseed and type of oilseed did not affect uterine inflammation at 25±1 d postpartum. Oilseed supplementation did not alter the intervals from calving to establishment of the first dominant follicle, preovulatory-size follicle, and ovulation, nor did it affect fertility (conception rate to first artificial insemination and proportion of pregnant cows by 150 d after calving). In summary, prepartum oilseed supplementation (6.2 to 7.4% ether extract, % of dietary dry matter) decreased DMI during the entire experimental period (pre- and postpartum), decreased milk yield during early lactation in multiparous cows, and increased calf birth weight with no significant improvement in ovarian function and reproductive performance., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective was to examine risk factors for the interval to resumption of ovarian cyclicity (ROC), multiple ovulations (MCL), ovarian follicular cysts (OC), and presence of intrauterine fluid (IUF) at 22 to 28 [visit (V) 1] and 36 to 42 (V2) days in milk (DIM) in dairy cows. The study was conducted retrospectively by evaluating records from 1,155 Holstein cows from 3 herds. Ovaries and uteri were examined at V1 and V2 by transrectal ultrasonography to determine ovarian structures and IUF. Based on the odds ratio, multiparous cows were more likely to have ROC at V1 by a factor of 1.79 compared with primiparous cows. The likelihood of ROC at V1 was lower in cows with higher milk production, in cows with retained fetal membranes (RFM) or cows with IUF at V1 by factors of 0.98 (for each kg of milk increased), 0.52, and 0.61, respectively. Based on the odds ratio, cows diagnosed with IUF at V2 were 2.85 times more likely to have attained ROC at V2. Multiparous cows and cows that delivered twins were 2.73 and 2.16 times, respectively, more likely to have MCL at V1, whereas cows with RFM were 0.38 times less likely to have MCL at V1. The likelihood of MCL at V2 was higher in cows with MCL and OC at V1 by factors of 2.67 and 1.91, respectively. Multiparous cows were 8.51 times more likely to have OC at V1 than primiparous cows. Higher producing cows were more likely to have OC at V2 by a factor of 1.04 compared with lower producing cows. Parity, stillbirth, RFM, and ROC at V1 were all identified as risk factors for IUF at V1. Cows with RFM and delivering twins were more likely to be diagnosed with IUF at V2 by a factor of 3.43 and 4.07, respectively. In summary, parity, twinning, RFM, metritis, IUF, and milk production were all associated with altered ovarian structures, and the presence of IUF was related to parity, twinning, RFM, and ROC in postpartum dairy cows., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Our objective was to evaluate the effects of dietary starch content, altered by partial substitution of dietary grain with wheat dried distillers grain with solubles (DDGS), on the interval from calving to first ovulation, concentrations of hormones and metabolites in plasma and follicular fluid, and granulosa cell gene expression in preovulatory follicles. Sixty lactating dairy cows were assigned to 1 of 2 diets from calving until 84d postpartum. Diets were formulated to contain either 17.3% rolled barley grain (29.2% starch) or 17.2% wheat DDGS (19.1% starch), with 43.0% barley silage and 21.6% rolled corn grain as the other major ingredients (dry matter basis). Transrectal ultrasonography was performed twice weekly to monitor ovarian dynamics from 7 ± 2d postpartum until ovulation or until 56d in milk, whichever occurred earlier. Plasma concentrations of insulin and insulin-like growth factor-1 (IGF-1) were determined in all 60 cows, and that of glucose, fatty acids, and urea in a subset of 24 cows, representing those in which the first ovulation occurred spontaneously within 5 wk postpartum. Estradiol (proestrus) and progesterone (12d postovulation) in plasma were also measured. Concentrations of insulin, IGF-1, glucose, fatty acids, and urea were determined in follicular fluid (wk 9), and the expression of LH receptor, estrogen receptor β, cytochrome P450 aromatase, and plasma type glutathione peroxidase genes measured in granulosa cells obtained from the preovulatory follicles at wk 9 postpartum in the subset of 24 cows. Diets did not alter the interval from calving to first ovulation (32.3 ± 2.5d), but a significantly lower proportion of cows on the DDGS diet (20%) ovulated multiple (≥ 2) follicles at the first ovulation than those on the barley grain diet (40%). The incidence of multiple ovulations tended to be lower at first insemination (10 vs. 21% for cows fed DDGS and barley grain diets, respectively). Mean plasma concentration of insulin was higher in cows fed the barley grain diet (2.5 vs 1.6 IU/mL), and a diet by time interaction was noted, with cows on the barley grain ration having higher insulin from wk 6 to 12 postpartum; however, mean plasma IGF-1 concentration did not differ between dietary groups. In the subsets, mean plasma concentrations of metabolites or estradiol and progesterone were not affected by diet, parity, or diet by parity interactions. Cows on the DDGS diet had lower concentrations of IGF-I (69 vs. 108 ng/mL) and higher fatty acids (222 vs. 149 mEq/L) in the follicular fluid obtained from preovulatory follicles. Diet, parity, and diet by parity interactions did not affect the concentrations of insulin, glucose, urea, estradiol, and progesterone in follicular fluid. Diets did not alter the expression profiles of LHr, estrogen receptor β, CYP19, and GPx3 genes in granulosa cells. In summary, diets did not affect the interval from calving to first ovulation or granulosa cell gene expression. However, reducing dietary starch content by a partial replacement of dietary grain with wheat DDGS increased fatty acids in follicular fluid and reduced the concentrations of insulin in plasma, IGF-1 in follicular fluid, and the incidence of multiple ovulations., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objectives of this study were to determine the effect of PGF2α treatment 14 d before the initiation of a G6G or G7G (PGF2α, 2 d, GnRH, 6 or 7 d, Ovsynch) protocol on ovarian response, synchronization protocol on ovarian response, progesterone (P4) concentration, pregnancy per AI (P/AI), and pregnancy loss in multiparous Holstein cows. Cows (3.6±1.1 lactations and yielding >30kg/d of milk) were randomly assigned to 1 of 4 timed artificial insemination (TAI) protocols: 1) G6G (n=240), one injection of PGF2α, GnRH 2 d later and a 7-d Ovsynch protocol (GnRH, 7 d, PGF2α, 56 h, GnRH 16 h TAI) was initiated 6 d later; 2) PG6G (n=250), PGF2α 14 d before the initiation of the G6G protocol; 3) G7G (n=200), one injection of PGF2α, GnRH 2 d later, and a 7-d Ovsynch protocol initiated 7 d later; and (4) PG7G (n=200), a PGF2α injection 14 d before the initiation of the G7G protocol. Blood samples from a subset of 269 cows were collected at the times of first and second GnRH, and PGF2α of the Ovsynch protocol to measure P4. Ultrasound examinations were performed to evaluate ovarian response to GnRH and PGF2α of Ovsynch, and to determine pregnancy status at 32 and 60 d after TAI. The proportion of cows with high (≥1ng/mL) P4 at first GnRH of Ovsynch was greater for PG6G and PG7G compared with G6G and G7G groups (combined 79.7 vs. 59.3%). In addition, mean (±SEM) plasma P4 concentration (ng/mL) at PGF2α of Ovsynch was also greater in PG6G (6.5±0.2) and PG7G (6.7±0.3) compared with G6G (5.1±0.2) and G7G (5.0±0.2). Cows given PGF2α 14 d before initiating a G6G or a G7G TAI (PG6G and PG7G) tended to have a greater P/AI at 32 d compared with those cows not receiving PGF2α (G6G and G7G). However, P/AI at 60 d was greater in cows subjected to PG6G and PG7G protocols (31.1 vs. 39.2%), with a lower pregnancy loss between 32 and 60 d (11.65 vs. 19.7%). In summary, administration of PGF2α 14 d before initiating a G6G or a G7G TAI protocol increased P4 concentrations before artificial insemination and late embryonic/early fetal survival in multiparous Holstein cows., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

A protocol for presynchronization of ovarian status with 2 injections of PGF2α given 14 d apart, with the last PGF2α injection given 12 or 14 d before Ovsynch increases pregnancy per artificial insemination (P/AI) in dairy cows. We determined the efficacy of reducing the interval from the last PGF2α injection (500 μg of cloprostenol) of presynchronization to initiation of Ovsynch on response to treatment and P/AI. Lactating dairy cows were assigned to an Ovsynch protocol, with the initial injection of GnRH given either 9 (PRE-9; n=135) or 12d (PRE-12; n=135) after the second PGF2α injection of presynchronization. The Ovsynch protocol consisted of 2 injections of 100 μg of GnRH given 9 d apart and 1 injection of PGF2α given 7 d after the initial GnRH injection, and cows were subjected to timed artificial insemination (TAI; 70±3.5 DIM) approximately 16 h after the second GnRH injection. Body condition score (1-5 scale) was recorded at TAI. Blood samples were taken for progesterone determination at the PGF2α injection of Ovsynch, at TAI, and at 11 d after TAI. Ultrasonographic examinations were done in all cows at the second PGF2α injection of presynchronization, initial GnRH injection, PGF2α injection of Ovsynch, at TAI, and 24 h after TAI for cyclicity status and ovarian responses to treatments, and at 32 and 60 d after TAI for confirmation of pregnancy. Overall, 29 cows (10.7%) were determined acyclic or cystic and excluded from the study. The percentage of cows responding to initial GnRH injection (62.2 vs. 61.5%) did not differ between PRE-9 and PRE-12 but more cows in the PRE-9 group failed to respond to PGF2α treatment of Ovsynch compared with PRE-12 (22.7 vs. 10.7%). Body condition score at TAI (2.9±0.02) and mean ovulatory follicle diameter (16.4±0.2 mm) were not different between treatments. Overall P/AI at 32 d was reduced in PRE-9 (33.6%) compared with PRE-12 (44.3%) but pregnancy losses (5.0 vs. 3.7%) did not differ between treatments. Primiparous cows in the PRE-12 group had higher mean progesterone concentration 11 d after TAI and greater P/AI 32 after TAI than primiparous cows in the PRE-9 group (6.4±0.5 vs. 4.6±0.5 ng/mL and 55.8 vs. 30.0%, respectively). In conclusion, reducing the interval from the last PGF2α injection of the presynchronization treatment to initiation of Ovsynch (from 12 to 9 d) did not affect ovulatory response to initial GnRH injection but reduced response to PGF2α injection of Ovsynch and P/AI at 32 and 60 d after TAI. The reduction in P/AI was particularly evident in primiparous cows of the PRE-9 group., (Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to examine the effects of dietary starch source and content in the immediate postpartum period on plasma metabolites and hormones and ovarian follicular development. One of 3 diets was fed in a randomized block design to 40 cows from calving until 70 d in milk. The diets contained 45% alfalfa silage (AS), 45% barley silage (BS), or 41% barley silage and 4% supplemental starch (SS) on a dry matter basis. All diets contained 45% barley-based concentrate and 10% alfalfa hay. Resulting starch levels were 25.2, 23.3, and 26.7% for AS, BS, and SS, respectively. Body condition was scored every other week and dry matter intake and milk yield were recorded daily. Milk samples were obtained weekly and blood samples were taken at calving and then every other week to determine concentrations of glucose, insulin, insulin-like growth factor-1, β-hydroxybutyrate, and nonesterified fatty acids. Transrectal ultrasonography was performed twice per week from 7 d after calving until first ovulation or 62 d in milk in all cows. For a subset of 7 AS-, 8 BS-, and 9 SS-fed cows, a complete estrous cycle was monitored for ovarian dynamics, and blood samples were collected every second day for progesterone and estradiol. Luteinizing hormone pulsatility was also determined (5 cows/treatment) approximately 15 d postcalving. Treatment had no effect on body condition score, dry matter intake, blood metabolites, milk yield, or milk fat and protein contents, but BS-fed cows had significantly higher levels of milk urea nitrogen compared with SS cows. Cows fed SS (31 d) tended to have a shorter interval from calving to first ovulation than cows fed AS (43 d) or BS (38 d). The incidence of double first ovulations was higher in cows fed SS (46%) compared with those fed BS (0%). Treatment had no effect on LH pulse frequency or amplitude, ovarian dynamics, or progesterone and estradiol concentrations during the observed estrous cycle. Energy balance did not differ among cows fed the 3 diets. Overall, dietary starch source and concentration had little effect on productivity or metabolic status of postpartum cows., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to determine the effects of feed restriction and source of dietary fatty acids during the close-up dry period on postcalving reproductive performance of dairy cattle. Thirty-four days before expected calving, pregnant Holstein cows (n = 72; parity 1 to 5) were randomly assigned to 1 of 6 treatments. Treatments were ad libitum (AL) or 24% feed restriction (FR) in combination with 1 of 3 oilseed supplements at 8% of diet dry matter: canola, linola, or flax to enrich the rations with oleic, linoleic, or linolenic fatty acids, respectively. After calving, cows were fed a common lactation diet that contained no oilseeds. Measurements of uterus, corpus luteum, and follicles were obtained by ultrasonography twice weekly from 7 +/- 1 d after calving until the first ovulation. Cows (n = 66) were subjected to timed artificial insemination (TAI), and pregnancy was determined 32 d later. Feed-restricted cows had lower dry matter intake and lost more body weight prepartum. Energy balance (Mcal/d) was negative in FR cows prepartum but they had a less severe negative energy balance postpartum. The dietary source of fatty acid did not affect energy balance. Cows fed AL had a higher incidence of uterine infections (10/37 vs. 2/35) but tended to have fewer ovarian cysts (2/37 vs. 7/35) than FR cows. Mean (+/-SE) interval from calving to uterine involution did not differ among dietary treatments (26.8 +/- 1.8 d). Interval from calving to first ovulation was longer in cows fed canola than in those fed either linola or flax (34.7 +/- 3.1 vs. 23.7 +/- 3.2 and 21.0 +/- 3.1 d, respectively). A greater percentage of cows fed AL conceived to the first TAI (47.1 vs. 18.8) and tended to have fewer mean days open (157 +/- 10.8 vs. 191 +/- 10.1) than cows fed FR. In summary, FR cows had a lower incidence of uterine infections, but they were less fertile as reflected by a lower percent pregnancy to first TAI and increased days open. Cows fed diets enriched in linoleic or linolenic fatty acids had a lesser incidence of ovarian cysts and ovulated sooner with no effect on energy balance or fertility.

The objective was to compare pregnancy rates in nulliparous Holstein heifers given PGF(2alpha) 24 h before, or concurrent with, removal of an intravaginal progesterone-releasing (CIDR) insert in 3 timed artificial insemination (TAI) protocols. Heifers (from 2 herds) were assigned randomly, over 11 mo, to 1 of 3 modified Ovsynch protocols. On d 0 (without reference to the stage of the estrous cycle), all heifers were given 100 microg of GnRH i.m. and a CIDR insert (containing 1.9 g of progesterone). In the PG-7/P4-8 protocol (n = 99), PGF(2alpha) was given on d 7, and CIDR inserts were removed on d 8. In the PG-7/P4-7 (n = 98) and PG-8/P4-8 (n = 102) protocols, PGF(2alpha) administration and CIDR removal occurred concurrently, on d 7 or 8, respectively. In all 3 protocols, a second GnRH treatment (100 microg) was given 48 h after PGF(2alpha) with TAI 16 to 20 h later. Blood samples were collected (subset of 124 heifers) on d 0, 7, 10 or 11 (i.e., at TAI), and 17. Pregnancy rates (32 d after TAI) for protocols PG-7/P4-8, PG-7/P4-7, and PG-8/P4-8 were 61.8, 55.6, and 54.1%, respectively. Pregnancy rate was higher when synchronization was initiated during diestrus than when initiated at other stages (57.0 versus 34.8%). Although pregnancy rates were not affected by season, there was an interaction between protocol and season; pregnancy rates were significantly lower in summer in heifers subjected to PG-7/P4-7 and PG-8/P4-8, but season did not affect pregnancy rates in heifers subjected to PG-7/P4-8. In summary, giving PGF(2alpha) 24 h before CIDR removal, followed by TAI (PG-7/P4-8 protocol), resulted in consistent pregnancy rates, regardless of season, relative to protocols involving PGF(2alpha) treatment concurrent with CIDR removal.