Postpartum vaginal discharge score is associated with genetic traits, postpartum fertility phenotypes, metabolic status, and overall reproductive performance in seasonal-calving pasture-based dairy cows.

The aims of this study were (1) to evaluate potential associations between genetic traits, postpartum phenotypes, cow factors, and postpartum vaginal discharge score (VDS); and (2) to investigate possible associations between postpartum VDS, plasma progesterone (P4) after first service, and reproductive performance. First- and second-parity (n = 2,842) spring-calving lactating dairy cows from 35 dairy herds were enrolled. Farm visits were performed every 2 wk during the postpartum period and weekly during the breeding period. Cows that were at wk 3 and wk 7 postpartum and between 7 and 13 d after first artificial insemination (AI) were examined. Body condition score (BCS) was measured on all farm visits using a 1-to-5 scale (low ≤2.5; target ≥2.75). Transrectal ultrasound examinations were conducted to determine the presence or absence of a corpus luteum (CL). Vaginal discharge score was determined at wk 3 and wk 7 using a Metricheck device (Simcro) and a 1-to-4 scale (1 = clear mucus; 4 = mucopurulent with >50% purulent material ± odor). At wk 3, cows having a VDS ≤2 were considered to have normal reproductive tract health status (RTHS). At wk 7, cows having VDS = 1 were considered to have normal RTHS. Blood samples were collected at each visit, and plasma concentrations of glucose, ß-hydroxybutyrate, fatty acids, and progesterone (only during breeding visit) were analyzed. Animals with target BCS at wk 3 and wk 7 had greater odds of having normal RTHS at wk 3 and wk 7, respectively, than cows with low BCS. Cows with a CL at wk 3 and wk 7 had greater prevalence of normal RTHS at wk 3 and wk 7, respectively, compared with cows without a CL. Cows with normal RTHS had a lesser plasma concentration of ß-hydroxybutyrate at wk 3 and wk 7 and greater plasma concentration of glucose at wk 3 compared with animals with abnormal RTHS. More cows in the greatest quartiles for the fertility subindex of the Economic Breeding Index and genetic merit for milk production traits (milk kg and milk protein [%]) had normal RTHS at wk 3 and wk 7 compared with the other quartiles. Cows with VDS = 4 at wk 7 postpartum had lesser plasma P4 concentration after first AI (-1.2, -1.1, and -1.0 ng/mL compared with cows with VDS = 1, 2, and 3, respectively). Similarly, cows with VDS = 4 at both wk 3 and wk 7 had lesser pregnancy at first service, lesser cumulative pregnancy rates at wk 3, 6, and 12 during the breeding period, and longer interval from mating start date to conception (+3 d if VDS = 4 at wk 3; +5 d if VDS = 4 at wk 7), compared with cows having other VDS. In conclusion, cows with superior genetic merit for fertility traits and milk production traits, and favorable fertility phenotypes at wk 3 and wk 7, were all associated with greater likelihood of having normal RTHS. In turn, abnormal postpartum RTHS and greater postpartum VDS (score = 4) were associated with lesser odds of successful pregnancy establishment.

Plasma progesterone concentration after first service is associated with individual genetic traits, postpartum phenotypes, and likelihood of conception in seasonal-calving pasture-based dairy cows.

The aims of this study were to (1) evaluate postpartum phenotypes, cow factors, and genetic traits associated with plasma progesterone (P4) concentrations after first artificial insemination (AI); (2) determine variation in daily plasma P4 concentrations between d 7 and 13 after first AI; and (3) evaluate associations between plasma P4 concentrations and pregnancy success after first AI. First and second parity (n = 2,797) spring-calving lactating dairy cows from 35 dairy herds were enrolled. Farm visits were performed every 2 wk during the postpartum period as follows: cows that were at wk 3 (range: 14-27 d in milk) and wk 7 (range: 42-55 d in milk) postpartum were examined. Farm visits were performed weekly during the breeding season, and cows that were between 7 and 13 d after the first AI were examined. Body condition score (BCS) was measured at each visit using a 1 to 5 scale [low (≤2.75), target (≥3.0)]. Transrectal ultrasound examinations were conducted at wk 3 and wk 7 postpartum visits to determine presence or absence of a corpus luteum (CL) and uterine tract score [scale of G1 (best)-G4 (worst)]. Blood samples were collected at each visit, and plasma concentrations of glucose, ß-hydroxybutyrate, and fatty acids were analyzed. On the day of the weekly farm visit during the breeding season, blood samples for P4 determination were collected from all cows that were between 7 and 13 d after first AI during the breeding period. Cows that had a CL present and a G1 uterine score at wk 7 postpartum had greater plasma P4 concentration after first AI (+0.67 ng/mL and +0.4 ng/mL, respectively) compared with cows with no CL present and with a uterine score ≥G3. Cows with low BCS at wk 7 postpartum had lesser plasma P4 concentration after first AI than cows with target BCS. Each unit increase in plasma fatty acids and ß-hydroxybutyrate concentration at AI was associated with 0.45 ± 0.33 ng/mL (estimate ± standard error) and 0.07 ± 0.04 ng/mL greater plasma P4 concentration after first AI, respectively. Regarding genetic merit traits, each unit increase in fertility subindex was associated with 0.005 ± 0.003 ng/mL greater P4 concentration. In addition, for every 1 ng/mL increase in plasma P4 concentration, the odds of estimated probability of pregnancy per AI increased by 3% (odds ratio = 1.03; 95% confidence interval = 1.00, 1.05). In conclusion, cows with superior genetic merit for fertility traits and milk production traits, favorable fertility phenotypes at wk 7 postpartum, (e.g., presence of a CL, a G1 uterine score, and target BCS), and blood parameters indicative of better metabolic status at AI were all associated with greater plasma P4 concentration after AI. In turn, greater plasma P4 concentrations were associated with greater odds of successful pregnancy establishment. This study underlines the important associations between early postpartum fertility phenotypes (CL presence, uterine health status) and subsequent plasma P4 concentrations after first AI, and hence provides additional evidence of the mechanisms through which selection for fertility traits improves phenotypic fertility performance.

Associations between postpartum fertility phenotypes and genetic traits in seasonal-calving, pasture-based lactating dairy cows.

The objective of this study was to evaluate the associations between corpus luteum (CL) status, uterine health, body condition score (BCS), metabolic status, and parity at wk 3 and 7 postpartum in seasonal-calving, pasture-based, lactating dairy cows. The associations between those phenotypes and individual genetic traits were also evaluated. First- and second-parity spring-calving lactating dairy cows (n = 2,600) from 35 dairy farms in Ireland were enrolled. Farms were visited every 2 weeks; cows that were at wk 3 (range 14 to 27 DIM) and wk 7 (range 42 to 55 DIM) postpartum were examined. Body condition score was measured using a scale of 1 to 5 with 0.25 increments. Transrectal ultrasound examination was performed at wk 3 and 7 postpartum to determine presence or absence of CL and ultrasound reproductive tract score. Blood samples were collected at each visit and the concentrations of glucose, ß-hydroxybutyrate (BHB), and fatty acids (FA) were analyzed by using enzymatic colorimetry. Cows were grouped into 3 BCS categories [low (≤2.5), target (≥2.75 and ≤3.25), and high (≥3.5)]; 2 CL status categories: (present or absent); 2 uterine health status (UHS) categories (normal and abnormal); and 3 metabolic status categories [good (high glucose, low fatty acids and BHB), poor (low glucose, high fatty acids and BHB), and moderate (all other combinations)]. Fisher's exact test was used to test associations between variables and was supplemented by logistic regression. We found associations between UHS (wk 3 and 7), BCS (wk 3 and 7), parity (wk 3 and 7) metabolic status (wk 3), and predicted transmitting ability for calving interval (PTA for CIV; wk 3) and CL status. Cows that had abnormal UHS, low BCS, primiparity, and poor metabolic status, and were in the quartile with the greatest PTA for CIV were less likely to have had CL present at wk 3 and 7 postpartum. We also found associations between CL status (wk 3 and 7), BCS (wk 3 and 7), parity (wk 3 and 7), and PTA for CIV (wk 3) and UHS. Cows that did not have a CL present had low BCS, primiparity, and that were in the quartile with greatest PTA for CIV, had a greater risk of abnormal UHS at wk 3 and 7 postpartum. We observed strong associations between CL status, UHS, BCS, metabolic status, parity, and individual genetic traits at wk 3 and 7 postpartum in seasonal-calving, pasture-based lactating dairy cows. Achieving target BCS and good metabolic status, and selecting cows based on PTA for CIV, are all expected to increase the likelihood of hastening the resumption of estrous cyclicity and enhancing uterine health during the postpartum period.

Associations between postpartum phenotypes, cow factors, genetic traits, and reproductive performance in seasonal-calving, pasture-based lactating dairy cows.

The objective of this study was to evaluate associations between corpus luteum (CL) status, uterine health, body condition score (BCS), metabolic status, parity, genetic merit for fertility traits, and reproductive performance in pasture-based dairy cows managed for seasonal reproduction. First- and second-lactation (n = 2,600) spring-calving dairy cows from 35 dairy farms located in Ireland were enrolled in the study. Farms were visited every 2 wk, and animals that were at wk 3 (range: 14-27 d in milk) and wk 7 (range: 42-55 d in milk) postpartum were examined. Body condition score was measured using a 1-to-5 scale in 0.25-point increments. Transrectal ultrasound examination was performed at wk 3 and 7 postpartum to determine presence or absence of CL and ultrasound reproductive tract score (scale of G1-G4). Blood samples were collected at each visit, and the concentrations of glucose, ß-hydroxybutyrate (BHB), and fatty acids (FA) were analyzed using enzymatic colorimetry. Animals were grouped into 3 BCS categories [low (≤2.5), target (2.75-3.25), and high (≥3.5)], 2 CL categories (present or absent), 2 uterine health status categories (normal or abnormal), and 3 metabolic status categories [good (high glucose, low FA and BHB), poor (low glucose, high FA and BHB), and moderate (all other combinations)]. Fisher's exact test was used to test for associations between variables and was supplemented by logistic regression. More cows with a CL at wk 7 were served during the first 21 d of the breeding period compared with cows without a CL. Cows classified as having a uterine score of G3 or G4 at wk 3 and 7 had lower odds of pregnancy establishment during the breeding period compared with animals with a uterine score of G1 or G2. Animals with low BCS at wk 7 had lower odds of pregnancy establishment than cows with a target BCS. Cows classified as having good metabolic status at both wk 3 and wk 7 had greater odds of pregnancy establishment during the first 21 d of the breeding season than those classified as having poor metabolic status. Overall, primiparous cows had greater reproductive performance than second-parity cows. Animals in the quartiles with the best predicted transmitting ability for survival and calving interval had better reproductive performance compared with animals in the other quartiles. Cows that had better genetic merit for fertility traits and good metabolic status, achieved target BCS, and had a favorable ultrasound reproductive tract score and a CL present at wk 7 postpartum had superior reproductive performance.

Anti-Müllerian hormone in grazing dairy cows: Identification of factors affecting plasma concentration, relationship with phenotypic fertility, and genome-wide associations.

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.

The effect of exogenous glucose infusion on early embryonic development in lactating dairy cows.

The objective of this study was to examine the effect of intravenous infusion of glucose on early embryonic development in lactating dairy cows. Nonpregnant, lactating dairy cows (n = 12) were enrolled in the study (276 ± 17 d in milk). On d 7 after a synchronized estrus, cows were randomly assigned to receive an intravenous infusion of either 750 g/d of exogenous glucose (GLUC; 78 mL/h of 40% glucose wt/vol) or saline (CTRL; 78 mL/h of 0.9% saline solution). The infusion period lasted 7 d and cows were confined to metabolism stalls for the duration of the study. Coincident with the commencement of the infusion on d 7 after estrus, 15 in vitro-produced grade 1 blastocysts were transferred into the uterine horn ipsilateral to the corpus luteum. All animals were slaughtered on d 14 to recover conceptuses, uterine fluid, and endometrial tissue. Glucose infusion increased circulating glucose concentrations (4.70 ± 0.12 vs. 4.15 ± 0.12 mmol/L) but did not affect milk production or dry matter intake. Circulating ß-hydroxybutyrate concentrations were decreased (0.51 ± 0.01 vs. 0.70 ± 0.01 mmol/L for GLUC vs. CTRL, respectively) but plasma fatty acids, progesterone, and insulin concentrations were unaffected by treatment. Treatment did not affect either uterine lumen fluid glucose concentration or the mRNA abundance of specific glucose transporters in the endometrium. Mean conceptus length, width, and area on d 14 were reduced in the GLUC treatment compared with the CTRL treatment. A greater proportion of embryos in the CTRL group had elongated to all length cut-off measurements between 11 and 20 mm (measured in 1-mm increments) compared with the GLUC treatment. In conclusion, infusion of glucose into lactating dairy cows from d 7 to d 14 post-estrus during the critical period of conceptus elongation had an adverse impact on early embryonic development.

Effect of manipulating progesterone before timed artificial insemination on reproductive and endocrine parameters in seasonal-calving, pasture-based Holstein-Friesian cows.

Fertility to timed AI (TAI) is profoundly affected by progesterone (P4) levels during hormonal synchronization protocols. Holstein-Friesian dairy cows managed in a seasonal-calving, pasture-based production system were randomly assigned to 2 treatments to manipulate P4 before TAI during growth of the preovulatory follicle. Cows in the first treatment (High P4; n=30) were submitted to a Double-Ovsynch protocol {Pre-Ovsynch [GnRH; 7 d, PGF2α; 3 d, GnRH] followed 7 d later by Breeding-Ovsynch [GnRH (G1); 7 d PGF2α; 24 h, PGF2α; 32 h, GnRH (G2); 16 h, TAI]}. Cows in the second treatment (n=30; Low P4) received the same Double-Ovsynch protocol but with an additional PGF2α treatment 24 h after G1. Overall, synchronization rate did not differ between treatments and was 92% (55/60). Unexpectedly, 37% of Low P4 cows were detected in estrus ~24 h before scheduled TAI and were inseminated ~16 h before scheduled TAI. Overall, P4 did not differ between treatments at G1, whereas High P4 cows had greater P4 concentrations at PGF2α and G2 than Low P4 cows. High P4 cows had the smallest mean follicle diameter at G2, whereas Low P4 cows with no estrus before TAI had intermediate mean follicle diameter at G2, and Low P4 cows with estrus before TAI had the largest mean follicle diameter. Low P4 cows with estrus before TAI had larger corpora lutea 15 d after TAI than Low P4 cows without estrus before TAI or High P4 cows. In accordance with corpus luteum size on d 15, High P4 cows and Low P4 cows without estrus before TAI had lower P4 from 4 to 46 d after TAI than Low P4 cows with estrus before TAI. Relative mRNA levels of the interferon-stimulated genes ISG15, MX1, MX2, and OAS1 were greater for Low P4 than for High P4 cows, whereas relative mRNA levels of RTP4 were greater for High P4 than for Low P4 cows 18 d after TAI. Treatment did not affect plasma pregnancy-associated glycoprotein concentrations after TAI; however, pregnancy-associated glycoprotein concentrations were affected by pregnancy status and parity. Treatment did not affect pregnancy per artificial insemination at 29, 39, or 60 d after TAI, and no pregnancy losses were observed from 39 to 60 d after TAI. We concluded that (1) Low P4 cows were more likely to express estrus than High P4 cows; (2) the subpopulation of Low P4 cows that expressed estrus had larger preovulatory follicles and greater P4 concentrations after TAI; and (3) regardless of estrus before TAI, all Low P4 cows had greater mRNA expression for 5 of 6 interferon-stimulated genes than High P4 cows 18 d after TAI.

Relationships between fertility and postpartum changes in body condition and body weight in lactating dairy cows.

The relationship between energy status and fertility in dairy cattle was retrospectively analyzed by comparing fertility with body condition score (BCS) near artificial insemination (AI; experiment 1), early postpartum changes in BCS (experiment 2), and postpartum changes in body weight (BW; experiment 3). To reduce the effect of cyclicity status, all cows were synchronized with Double-Ovsynch protocol before timed AI. In experiment 1, BCS of lactating dairy cows (n = 1,103) was evaluated near AI. Most cows (93%) were cycling at initiation of the breeding Ovsynch protocol (first GnRH injection). A lower percentage pregnant to AI (P/AI) was found in cows with lower (≤ 2.50) versus higher (≥ 2.75) BCS (40.4 vs. 49.2%). In experiment 2, lactating dairy cows on 2 commercial dairies (n = 1,887) were divided by BCS change from calving until the third week postpartum. Overall, P/AI at 70-d pregnancy diagnosis differed dramatically by BCS change and was least for cows that lost BCS, intermediate for cows that maintained BCS, and greatest for cows that gained BCS [22.8% (180/789), 36.0% (243/675), and 78.3% (331/423), respectively]. Surprisingly, a difference existed between farms with BCS change dramatically affecting P/AI on one farm and no effect on the other farm. In experiment 3, lactating dairy cows (n = 71) had BW measured weekly from the first to ninth week postpartum and then had superovulation induced using a modified Double-Ovsynch protocol. Cows were divided into quartiles (Q) by percentage of BW change (Q1 = least change; Q4 = most change) from calving until the third week postpartum. No effect was detected of quartile on number of ovulations, total embryos collected, or percentage of oocytes that were fertilized; however, the percentage of fertilized oocytes that were transferable embryos was greater for cows in Q1, Q2, and Q3 than Q4 (83.8, 75.2, 82.6, and 53.2%, respectively). In addition, percentage of degenerated embryos was least for cows in Q1, Q2, and Q3 and greatest for Q4 (9.6, 14.5, 12.6, and 35.2% respectively). In conclusion, for cows synchronized with a Double-Ovsynch protocol, an effect of low BCS (≤ 2.50) near AI on fertility was detected, but change in BCS during the first 3 wk postpartum had a more profound effect on P/AI to first timed AI. This effect could be partially explained by the reduction in embryo quality and increase in degenerate embryos byd 7 after AI in cows that lost more BW from the first to third week postpartum.

Effect of timing of initiation of resynchronization and presynchronization with gonadotropin-releasing hormone on fertility of resynchronized inseminations in lactating dairy cows.

Lactating Holstein cows (n=1,456) were randomized in a 2×2 factorial design to compare the main effects of day of initiation of resynchronization after artificial insemination (AI; 32 vs. 39 d) and presynchronization with GnRH 7d before initiation of resynchronization on fertility to timed AI (TAI). This design resulted in the following 4 resynchronization treatments: (1) resynchronization (GnRH treatment, PGF2α treatment 7d later, GnRH treatment 56 h later, and TAI 16 h later), initiated 32 ± 3 d after AI; (2) presynchronization with 100 µg of GnRH 25 ± 3 d after AI and resynchronization initiated 32 ± 3 d after AI at nonpregnancy diagnosis; (3) resynchronization initiated 39 ± 3 d after AI (GPG39); and (4) presynchronization with 100 µg of GnRH 32 ± 3 d after AI at nonpregnancy diagnosis and resynchronization initiated 39 ± 3 d after AI. Overall, 344 cows were inseminated at estrus between enrollment (25 ± 3 d after AI) and TAI of the resynchronization treatments, and 1,112 cows received TAI. Progesterone (P4) was analyzed in blood samples collected from all cows at the first GnRH injection of the resynchronization protocols (G1), and ovarian structures were evaluated and blood samples were collected at G1, at the PGF2α injection, and at the TAI of the resynchronization protocols in a subgroup of cows (n=417). When analyzed as main effects, cows presynchronized with GnRH had more pregnancies per AI (P/AI) than nonpresynchronized cows (38.9 vs. 33.8%), whereas timing of initiation of resynchronization did not affect P/AI. Although cows with high P4 at G1 had greater P/AI than cows with low P4 (38.7 vs. 31.8%), presynchronization with GnRH did not increase the proportion of cows with high P4 (>1.0 ng/mL) at G1 but moved cows from a low-P4 environment to an intermediate-P4 level. Presynchronization with GnRH also decreased the percentage of cows with low P4 at the PGF2α injection, thereby increasing synchrony to the protocol. Cows with high P4 at G1 had a decreased ovulatory response to G1 compared with cows with low P4 (40.9 vs. 69.1%), and cows that ovulated to G1 had decreased luteal regression after PGF2α compared with cows that did not ovulate (78.5 vs. 87.3%). We conclude that presynchronization with GnRH 7d before initiation of resynchronization increased fertility in dairy cows, whereas timing of initiation of resynchronization did not.

Factors associated with fertility outcomes in cows treated with protocols to synchronize estrus and ovulation in seasonal-calving, pasture-based dairy production systems.

Logistic regression was used to identify factors associated with fertility outcomes in cows treated with protocols to synchronize estrus and ovulation. Lactating dairy cows (n=1,538) were enrolled in a completely randomized block design study to evaluate synchronization treatments. Within each herd (n=8), cows were divided into 3 calving groups: early [≥ 42 d in milk (DIM) at mating start date (MSD); n=1,244], mid (21 to 41 DIM at MSD; n=179), and late (0 to 20 DIM at MSD; n=115), based on DIM at MSD. Cows in the early-, mid-, and late-calving groups were synchronized to facilitate estrus or timed artificial insemination (TAI) at MSD (planned breeding 1; PB1), 21 d (PB2), and 42 d (PB3) after MSD, respectively. For each PB, cows in the relevant calving group were stratified by parity and calving date and randomly assigned to (1) d -10 GnRH (10 µg i.m. of buserelin) and CIDR [controlled internal drug release insert, 1.38 g of progesterone (P4)]; d -3 PGF(2α) (25mg i.m. of dinoprost); d -2 CIDR out and AI at observed estrus (CIDR_OBS); (2) same as CIDR_OBS, but GnRH 36 h after CIDR out and TAI 18 h later (CIDR_TAI); (3) same as CIDR_TAI, but no CIDR (i.e., Ovsynch); or (4) untreated controls (CTRL). Use of a CIDR-based ovulation synchronization protocol (i.e., CIDR_TAI) increased synchronization rates in anovular cows. Both CIDR_OBS and CIDR_TAI animals without a corpus luteum (CL) had increased likelihood of conception at first service compared with Ovsynch animals without a CL. Animals with low body condition score (BCS) treated with CIDR_OBS had an increased likelihood of conceiving at first service compared with low-BCS animals treated with CIDR_TAI, Ovsynch, or CTRL. Animals <60 d in milk (DIM) treated with CIDR_OBS and CIDR_TAI had increased likelihood of conceiving at first service compared with animals treated with Ovsynch. Treatment with CIDR_TAI increased synchronization rate in cows categorized as low BCS, anovulatory, and <60 DIM compared with both CIDR_OBS and Ovsynch, and increased submission rate compared with CIDR_OBS. Conception rate in cows within these categories, however, was greatest for CIDR_OBS, resulting in minimal differences in actual pregnancy rates between CIDR_OBS and CIDR_TAI treatments, both of which were superior to Ovsynch. Treatment differences in the response variables investigated were minimal in cows categorized as medium or high BCS, ovulatory, and >60 DIM, indicating that CIDR-based protocols could be targeted at particular cows, and all other cows could be synchronized using Ovsynch.

Presynchronization with Double-Ovsynch improves fertility at first postpartum artificial insemination in lactating dairy cows.

The objective of this study was to compare circulating progesterone (P4) profiles and pregnancies per AI (P/AI) in lactating dairy cows bred by timed artificial insemination (TAI) following Ovsynch-56 after 2 different presynchronization protocols: Double-Ovsynch (DO) or Presynch-Ovsynch (PS). Our main hypothesis was that DO would increase fertility in primiparous cows, but not in multiparous cows. Within each herd (n=3), lactating dairy cows (n=1,687; 778 primiparous, 909 multiparous) were randomly assigned to DO [n=837; GnRH-7d-PGF(2α)-3d-GnRH-7d-Ovsynch-56 (GnRH-7d-PGF(2α)-56h-GnRH-16hTAI)] or PS (n=850; PGF(2α)-14d-PGF(2α)-12d-Ovsynch-56). In 1 herd, concentrations of P4 were determined at the first GnRH (GnRH1) of Ovsynch-56 and at d 11 after TAI (n=739). In all herds, pregnancy was diagnosed by palpation per rectum at 39 d. In 1 herd, the incidence of late embryo loss was determined at 74d, and data were available on P/AI at the subsequent second service. Presynchronization with DO reduced the percentage of animals with low P4 concentrations (<0.50 ng/mL) at GnRH1 of Ovsynch-56 (5.4 vs. 25.3%, DO vs. PS). A lesser percentage of both primiparous and multiparous cows treated with DO had low P4 concentrations at GnRH1 of Ovsynch-56 (3.3 vs. 19.7%, DO vs. PS primiparous; and 8.8 vs. 31.9%, DO vs. PS multiparous). Presynchronization with DO improved P/AI at the first postpartum service (46.3 vs. 38.2%, DO vs. PS). Statistically, a fertility improvement could be detected for primiparous cows treated with DO (52.5 vs. 42.3%, DO vs. PS, primiparous), but only a tendency could be detected in multiparous cows (40.3 vs. 34.3%, DO vs. PS, multiparous), consistent with our original hypothesis. Presynchronization treatment had no effect on the incidence of late embryo loss after first service (8.5 vs. 5.5%, DO vs. PS). A lower body condition score increased the percentage of cows with low P4 at GnRH1 of Ovsynch-56 and reduced fertility to the TAI. In addition, P4 concentration at d 11 after TAI was reduced by DO. The method of presynchronization at first service had no effect on P/AI at the subsequent second service (34.7 vs. 36.5%, DO vs. PS). Thus, presynchronization with DO induced cyclicity in most anovular cows and improved fertility compared with PS, suggesting that DO could be a useful reproductive management protocol for synchronizing first service in commercial dairy herds.

Effect of presynchronization with human chorionic gonadotropin or gonadotropin-releasing hormone 7 days before resynchronization of ovulation on fertility in lactating dairy cows.

Our objectives were to (1) compare the effect on pregnancies per artificial insemination (P/AI) of presynchronization of the estrous cycle with human chorionic gonadotropin (hCG) 7d before resynchronization of ovulation (Resynch) initiated 25 d after timed artificial insemination (TAI) and compare the presynchronization treatment with the Double-Ovsynch (DO) protocol, and (2) evaluate whether hCG for presynchronization could be replaced with GnRH. In experiment 1, lactating Holstein cows were blocked by parity and were randomly assigned to receive (1) Resynch-25 (D25), the Resynch protocol (GnRH-7 d-PGF(2 α)-56 h-GnRH-16 h-TAI) initiated 25 d after TAI (n=418); (2) HGPG, presynchronization with hCG (2,000 IU of Chorulon) 7d before D25 (n=450); and (3) DO (Pre-Resynch, GnRH-7 d-PGF(2 α)-72 h-GnRH; Breeding-Resynch, GnRH-7 d-PGF(2 α)-56 h-GnRH-16 h-TAI) initiated 22d after TAI (n=405). At 29 d after TAI, cows in the HGPG (37.3%) and DO (35.8%) groups had more P/AI than did cows in the D25 group (28.0%), and cows in the HGPG and DO groups continued to have more P/AI than did cows in the D25 group at 53 d after TAI. Presynchronization with hCG induced ovulation in 76% of the cows, which increased the percentage of HGPG cows with a corpus luteum at the initiation of Resynch compared with cows in the D25 group. In experiment 2, the D25 (n=368) and HGPG (n=338) treatments described in experiment 1 were compared in addition to a third treatment (GGPG; n=351), in which the hCG injection 18 d after TAI was replaced with a GnRH injection (200 µg of gonadorelin). At 32 d after TAI, cows in the HGPG group had more P/AI than did cows in the D25 group (33.7 vs. 25.5%), whereas cows in the GGPG group had intermediate P/AI (31.6%). At 53 d after TAI, P/AI tended to be greater for cows in the HGPG group than for those in the D25 group, whereas P/AI for cows in the GGPG group did not differ from that for cows in the D25 group. Treatment with hCG and GnRH 18d after TAI induced ovulation in 58.8 and 48.2% of cows, respectively, but did not increase the percentage of cows with a corpus luteum at the initiation of Resynch. More cows in the HGPG and GGPG groups had their estrous cycles synchronized after the resynchronization protocols compared with cows in the D25 group. We conclude that presynchronization with hCG increased fertility by increasing synchronization to the Resynch protocol, whereas presynchronization with GnRH improved synchronization to the Resynch protocol but did not improve fertility when compared with no presynchronization or presynchronization with hCG.

Effect of progesterone on magnitude of the luteinizing hormone surge induced by two different doses of gonadotropin-releasing hormone in lactating dairy cows.

Ovulation to the first GnRH injection of Ovsynch-type protocols is lower in cows with high progesterone (P4) concentrations compared with cows with low P4 concentrations, suggesting that P4 may suppress the release of LH from the anterior pituitary after GnRH treatment. The objectives of this study were to determine the effect of 1) circulating P4 concentrations at the time of GnRH treatment on GnRH-induced LH secretion in lactating dairy cows and 2) increasing the dose of GnRH from 100 to 200 µg on LH secretion in a high- and low-P4 environment. A Double-Ovsynch (Pre-Ovsynch: GnRH, PGF(2α) 7d later, GnRH 3d later, and Breeding-Ovsynch 7d later: GnRH, PGF(2α) 7d later, and GnRH 48 h later) synchronization protocol was used to create the high- and low-P4 environments. At the first GnRH injection of Breeding-Ovsynch (high P4), all cows with a corpus luteum ≥ 20 mm were randomly assigned to receive 100 or 200 µg of GnRH. At the second GnRH injection of Breeding-Ovsynch (low P4) cows were again randomized to receive 100 or 200 µg of GnRH. Blood samples were collected every 15 min from -15 to 180 min after GnRH treatment, and then hourly until 6h after GnRH treatment. As expected, mean P4 concentrations were greater for cows in the high- than the low-P4 environment. For cows receiving 100 µg of GnRH, the LH peak and area under the curve (AUC) were greater in the low- than in the high-P4 environment. Similarly, for cows receiving 200 µg of GnRH, the LH peak and AUC were greater in the low- than the high-P4 environment. Cows receiving 100 or 200 µg of GnRH had greater mean LH concentration in the low- than the high-P4 environment from 1 to 6h after GnRH treatment. On the other hand, when comparing the effect of the 2 GnRH doses in the high- and low-P4 environments, cows receiving 200 µg of GnRH had a greater LH peak and AUC than cows treated with 100 µg of GnRH both in the high- and low-P4 environments. For the high-P4 environment, mean LH was greater from 1.5 to 5h after GnRH treatment for cows receiving 200 µg of GnRH than for those receiving 100 µg of GnRH. In the low-P4 environment, mean LH was greater for cows receiving 200 µg of GnRH than for those receiving 100 µg of GnRH from 1 to 2.5h after GnRH treatment. We conclude that the P4 environment at GnRH treatment dramatically affects GnRH-induced LH secretion, and that a 200-µg dose of GnRH can increase LH secretion in either a high- or a low-P4 environment.

Effects of synchronization treatments on ovarian follicular dynamics, corpus luteum growth, and circulating steroid hormone concentrations in lactating dairy cows.

Lactating dairy cows (n=57) ≥45 d postpartum at first service were enrolled in a randomized complete block design study to evaluate treatments to synchronize estrus and ovulation. At 10 d before artificial insemination (AI), animals were randomly assigned to 1 of 3 treatments: (1) d -10 GnRH (GnRH1; 10 µg of buserelin, i.m.) and controlled internal drug release insert [CIDR, 1.38 g of progesterone (P4)]; d -3 PGF(2α) (PGF; 25 mg of dinoprost, i.m.); d -2 CIDR out; and AI at observed estrus (CIDR_OBS); (2) same as CIDR_OBS, but GnRH (GnRH2) 36 h after CIDR out and timed AI (TAI) 18 h later (CIDR_TAI); or (3) same as CIDR_TAI, but no CIDR (Ovsynch). Transrectal ultrasound was used to assess follicle size before ovulation and on d 4, 8, and 15 after the presumptive day of estrus (d 0) to measure the corpus luteum (CL). Blood samples were collected to determine concentrations of estradiol (E2; d -10, -9, -3, -2, -1, and 0) and P4 (d -10, -9, -2, -1, 0, 1, 4, 6, 8, 11, and 15). No treatment differences were observed in either circulating concentrations of P4 or the ovulatory response to GnRH1 at the onset of synchronization treatments. Circulating concentrations of P4 were greater for CIDR_OBS and CIDR_TAI compared with Ovsynch at 24 h after CIDR insertion (5.34 and 4.98 vs. 1.75 ng/mL) and immediately before CIDR removal (1.65 and 1.48 vs. 0.40 ng/mL). Peak circulating concentrations of E2 were greater for CIDR_OBS compared with Ovsynch (3.85 vs. 2.39 pg/mL), but CIDR_TAI (2.82 pg/mL) did not differ from either CIDR_OBS or Ovsynch. The interval from PGF injection to peak circulating E2 did not differ between CIDR_TAI and Ovsynch (52.1 vs. 49.8 h). Both CIDR_TAI and Ovsynch, however, had shorter intervals from PGF injection to peak circulating E2 concentrations compared with CIDR_OBS (67.8 h). The diameter of the dominant follicle before ovulation was greater for CIDR_OBS compared with Ovsynch (18.5 vs. 16.0 mm) but CIDR_TAI (17.1 mm) did not differ from either of the other treatments. The mean interval from PGF to ovulation was longer for CIDR_OBS (100.0 h) compared with CIDR_TAI and Ovsynch (84.4 and 83.2 h, respectively). Use of CIDR_OBS resulted in increased preovulatory follicle size and greater circulating concentrations of E2 due to a longer period of preovulatory follicle growth. Progesterone supplementation during synchronization and GnRH on the day before TAI affected ovulatory follicle size, and periovulatory circulating concentrations of P4 and E2. No differences, however, in postovulatory P4 or luteal volume profiles were observed.

Managing the dominant follicle in lactating dairy cows.

Reproductive efficiency is not optimal in high-producing dairy cows. Although many aspects of ovarian follicular growth in cows are similar to those observed in heifers, there are numerous specific differences in follicular development that may be linked with changes in reproductive physiology in high-producing lactating dairy cows. These include: 1) reduced circulating estradiol (E2) concentrations near estrus, 2) ovulation of follicles that are larger than the optimal size, 3) increased double ovulation and twinning, and 4) increased incidence of anovulation with a distinctive pattern of follicle growth in anovular dairy cows. The first three changes become more dramatic as milk production increases, although anovulation has not generally been associated with level of milk production. To overcome reproductive inefficiencies in dairy cows, reproductive management programs have been developed to synchronize ovulation and enable the use of timed AI in lactating dairy cows. Effective regulation of the CL, follicles, and hormonal environment during each part of the protocol is critical for optimizing these programs. This review discusses the distinct aspects of follicular development in lactating dairy cows and the methodologies that have been utilized in the past two decades in order to manage the dominant follicle during synchronization of ovulation and timed AI programs.

Evaluation of protocols to synchronize estrus and ovulation in seasonal calving pasture-based dairy production systems.

Lactating dairy cows (n=1,538) were enrolled in a randomized complete block design study to evaluate protocols to synchronize estrus and ovulation. Within each herd (n=8), cows were divided into 3 calving groups: early, mid, and late, based on days in milk (DIM) at mating start date (MSD). Early calving cows (n=1,244) were ≥42 DIM at MSD, mid-calving cows (n=179) were 21 to 41 DIM at MSD, and late-calving cows (n=115) were 0 to 20 DIM at MSD. Cows in the early, mid-, and late-calving groups were synchronized to facilitate estrus or timed AI (TAI) at MSD (planned breeding 1; PB1), 21 d (PB2), and 42 d (PB3) after MSD, respectively. For each PB, cows in the relevant calving group were stratified by parity and calving date and randomly assigned to 1 of 4 experimental groups: (1) d -10 GnRH (10 µg of i.m. buserelin) and controlled internal drug release insert (CIDR; 1.38 g of progesterone); d -3 PGF(2α) (25 mg of i.m. dinoprost); and d -2 CIDR out and AI at observed estrus (CIDR_OBS); (2) same as CIDR_OBS, but GnRH 36 h after CIDR out and TAI 18 h later (CIDR_TAI); (3) same as CIDR_TAI, but no CIDR (Ovsynch); or (4) untreated controls (CTRL). The CIDR_OBS, CIDR_TAI, and Ovsynch had shorter mean intervals from calving to first service compared with the CTRL (69.2, 63.4, and 63.7 vs. 73.7 d, respectively). Both CIDR_OBS (predicted probability; PP of pregnancy=0.59) and CIDR_TAI (PP of pregnancy=0.54) had increased odds of conceiving at first service compared with Ovsynch [PP of pregnancy=0.45; odds ratio (OR)=1.81 and OR=1.46, respectively], and Ovsynch had decreased likelihood of conceiving at first service (OR=0.70) compared with CTRL (PP of pregnancy=0.53). Both CIDR_TAI hazard ratio; HR [95% confidence interval=1.21 (1.04, 1.41)] and Ovsynch [HR (95% confidence interval)=1.23 (1.05, 1.44)] were associated with an increased likelihood of earlier conception compared with the CTRL. A greater proportion of cows on the CIDR_TAI treatment successfully established pregnancy in the first 42 d of the breeding season compared with the CTRL (0.75 vs. 0.67 PP of 42-d pregnancy, respectively). Protocols to synchronize estrus and ovulation were effective at achieving earlier first service and conception in pasture-based seasonal calving dairy herds. However, animals that conceived following insemination at observed estrus had a decreased likelihood of embryo loss to first service compared with animals bred with TAI (PP of embryo loss after first service=0.05 vs. 0.09; OR=0.52).