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.

Fertility of fresh and frozen sex-sorted semen in dairy cows and heifers in seasonal-calving pasture-based herds.

Our objective in this study was to evaluate the reproductive performance of dairy heifers and cows inseminated with fresh or frozen sex-sorted semen (SS) in seasonal-calving pasture-based dairy herds. Ejaculates of 10 Holstein-Friesian bulls were split and processed to provide (1) fresh conventional semen at 3 × 106 sperm per straw (CONV); (2) fresh SS at 1 × 106 sperm per straw (SS-1M); (3) fresh SS semen at 2 × 106 sperm per straw (SS-2M); and (4) frozen SS at 2 × 106 sperm per straw (SS-FRZ). Generalized linear mixed models were used to evaluate the effect of semen treatment and other explanatory variables on pregnancy per artificial insemination (P/AI) in heifers (n = 3,214) and lactating cows (n = 5,457). In heifers, P/AI was greater for inseminations with CONV (60.9%) than with SS-FRZ (52.8%) but did not differ from SS-1M (54.2%) or SS-2M (53.5%). Cows inseminated with CONV had greater P/AI (48.0%) than cows inseminated with SS, irrespective of treatment (SS-1M, SS-2M, and S-FROZEN; 37.6, 38.9, and 40.6%, respectively). None of the SS treatments differed from each other with regard to P/AI in either heifers or cows. The relative performance of SS compared with CONV was also examined [i.e., relative P/AI = (SS P/AI)/(CONV P/AI) × 100]. Frozen SS achieved relative P/AI >84%. Bull affected P/AI in both heifers and cows, but no bull by semen treatment interaction was observed. In heifers, P/AI increased with increasing predicted transmitting ability for milk protein percentage. In cows, P/AI increased with increasing Economic Breeding Index (EBI) and with days in milk (DIM) at AI but decreased with increasing EBI milk subindex, parity and with DIM2. Cows in parity ≥5 had the lowest P/AI and differed from cows in parities 1, 2, or 3. Dispatch-to-AI interval of fresh semen did not affect P/AI in lactating cows, but a dispatch-to-AI interval by bull interaction was detected whereby P/AI was constant for most bulls but increased with greater dispatch-to-AI intervals for 2 bulls. In conclusion, frozen SS achieved greater P/AI relative to conventional semen than was previously reported in lactating cows. Fresh SS did not achieve greater P/AI than frozen SS, regardless of whether the sperm dose per straw was 1 × 106 or 2 × 106. A bull effect for all semen treatments, as well as a dispatch-to-AI interval by bull interaction for fresh semen, highlights the importance of using a large team of bulls for breeding management.

Evaluation of alternative strategies to treat anoestrous dairy cows and implications for reproductive performance in pasture-based seasonal calving herds: A pilot study.

The objective of the present study was to assess the effects on ovulation and reproductive performance of a single injection of either GnRH or hCG applied 9 days before the start of the seasonal breeding period in anovulatory anoestrus cows compared with a 7-day progesterone-Ovsynch protocol. The study was conducted on four grass-based seasonal calving dairy herds in Ireland. The total number of cows in the herds was 2112, of which 488 were diagnosed as anoestrus based on absence of behavioural oestrus during a 30 day period. Ovarian structures and the uterus were examined by transrectal ultrasound on all 488 presumptive anestrus cows 9 days before mating start date (MSD). The number of corpora lutea (CL), number of large follicles (≥10 mm) and uterine reproductive tract score were recorded. Only cows that had no CL, ultrasound reproductive tract score ≤2 and were ≥30 days in milk (DIM) were enrolled in the study (n = 214). Cows were blocked by parity, DIM and body condition score and randomly assigned to one of four treatments: i.m. injection of gonadotropin releasing hormone analogue [GnRH; (n = 57)], i.m. injection of human chorionic gonadotropin [hCG; (n = 48)], 7-day Progesterone-Ovsynch protocol [P4OV; (n = 60)] and Control (no hormonal intervention, n = 49). A second ultrasound examination was performed 7 days after treatment to determine ovulatory response. There was a treatment effect on ovulation rate (P < 0.0001), whereby Control cows had a lesser ovulation rate compared with GnRH-, hCG- and P4OV-treated cows. Submission rate during the first 21 days of the breeding period [SR21; (P = 0.74)], pregnant to first service [P/AI1; (P = 0.24)], pregnant within 42 days after the onset of breeding [P42; (P = 0.73)], and pregnant within 84 days after the onset of breeding were not affected by treatment. A tendency was observed (P = 0.07) for greater likelihood of pregnancy within 21 days after the onset of breeding (P21) for P4OV and Control cows compared with GnRH- and hCG-treated cows. GnRH- and hCG-treated cows tended (P = 0.10) to have greater P/AI1 when first service events occurred after day 21 of the breeding period compared with Control cows. P4OV cows had shorter MSD to first service interval (P = 0.0001) and shorter MSD to conception interval (P = 0.02) compared with Control, GnRH- and hCG-treated cows. In conclusion, treatment of anestrous cows with GnRH or hCG resulted in an increase in ovulation rate compared with untreated Control cows, but did not improve reproductive performance during the first 21 days of the breeding season. The best reproductive performance results were obtained with the P4OV treatment, but this treatment has the greatest cost, and has the greatest number of interventions. The observation of good P/AI1 in hCG- and GnRH- treated cows when the first insemination occurred later than day 21 after MSD warrants further investigation, and suggests that these interventions should be applied earlier than 9 days before the farm MSD.

The relationship between anogenital distance and fertility, and genome-wide associations for anogenital distance in Irish Holstein-Friesian cows.

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; R2 = 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.

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.