Fertility in seasonal-calving pasture-based lactating dairy cows following timed artificial insemination or timed embryo transfer with fresh or frozen in vitro-produced embryos.

The objective was to compare pregnancy per service event (P/S) in lactating dairy cows following timed artificial insemination (AI) or timed embryo transfer (ET) using either fresh or frozen in vitro-produced embryos. Oocytes were collected once per week for up to 9 wk using transvaginal ovum pick-up from elite dairy donors (ET-DAIRY; n = 40; Holstein-Friesian and Jersey) and elite beef donors (ET-ELITE-BEEF; n = 21; Angus). Both ET-DAIRY and ET-ELITE-BEEF donors consisted of heifers and cows. In addition, oocytes were collected from the ovaries of beef heifers of known pedigree following slaughter at a commercial abattoir (ET-COMM-BEEF; n = 119). Following in vitro maturation and fertilization, presumptive zygotes were cultured in vitro to the blastocyst stage. Grade 1 blastocysts were either transferred fresh or frozen for on-farm thawing and direct transfer. A total of 1,106 recipient cows (all lactating, predominantly Holstein-Friesian) located on 16 herdlets were blocked based on parity, calving date, and Economic Breeding Index, and randomly assigned to receive AI (n = 243) or ET (n = 863) after estrous synchronization with a 10-d Progesterone-synch protocol. Cows assigned to ET were further randomized to receive fresh (n = 187) or frozen (n = 178) ET-ELITE-BEEF embryos, fresh (n = 169) or frozen (n = 162) ET-DAIRY embryos, or fresh (n = 80) or frozen (n = 87) ET-COMM-BEEF embryos. Pregnancy was diagnosed using transrectal ultrasound on d 32 to 35 after synchronized ovulation and confirmed on d 62 to 65, at which time fetal sex was determined. Pregnancy per service event at d 32 was not different between AI (48.8%) and ET (48.9%) and did not differ between dairy and beef embryos (50.3% vs. 48.1%, respectively). However, P/S was less on d 32 following transfer of frozen embryos (41.6%) compared with fresh embryos (56.1%). Pregnancy loss between d 32 and 62 was greater for ET (15.1%) compared with AI (4.7%), with greater losses observed for frozen beef (18.5%), fresh beef (17.3%), and frozen dairy (19.2%) compared with fresh dairy (6.0%) embryos. Serum progesterone (P4) concentration on d 7 was associated with P/S at d 32 and 62. Cows in the quartile with the least serum P4 concentrations (quartile 1) had less probability of being pregnant on d 32 (33.4%) compared with cows in the 3 upper quartiles for serum P4 (45.7%, 55.6%, and 61.2% for quartile 2, quartile 3, and quartile 4, respectively). Sex ratio (male:female) at d 62 was skewed toward more male fetuses following ET (61.1:38.9) compared with AI (43.2:56.8) and was consistent with the sex ratio among in vitro blastocysts (61.2:38.8). In conclusion, P/S was similar for AI and ET, although pregnancy loss between d 32 and 62 was greater for ET than for AI.

Effect of delayed timing of artificial insemination with sex-sorted semen on pregnancy per artificial insemination in synchronized dairy heifers managed in a seasonal-calving pasture-based system.

The objective of this study was to evaluate the timing of artificial insemination (AI) with frozen-thawed sex-sorted semen on pregnancy per AI (P/AI) in dairy heifers. A 6-d progesterone Co-Synch protocol was used for ovulation synchronization of dairy heifers, with timed AI (TAI) coincident with (TAI-0) or 8 h (TAI-8) after the second injection of GnRH, corresponding to either 48 h or 56 h after removal of the progesterone-releasing intravaginal device. Pregnancy diagnosis was conducted by transrectal ultrasound scanning of the uterus 34 d after TAI (n = 816 records available for analysis). Generalized linear mixed models were used to examine the effects of treatment on P/AI. Treatment (n = 2), herd (n = 11), and treatment × herd were included as categorical fixed effects. Heifer body weight and Economic Breeding Index values for milk production, fertility, calving performance, beef carcass, cow maintenance, cow management, and health were included as continuous fixed effects. Heifer ID was included as a random effect. Pregnancy per AI was greater for TAI-8 heifers (59%) compared with TAI-0 heifers (50%). Pregnancy per AI ranged from 38% to 75% between herds but there was no treatment × herd interaction. The fertility subindex (positive) and the cow management subindex (negative) were the only continuous animal variables associated with P/AI. Delaying the timing of AI with frozen-thawed sex-sorted semen by 8 h in dairy heifers enrolled on a 6-d progesterone Co-Synch protocol improved P/AI.

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.

Associations between the postpartum uterine and vaginal microbiota and the subsequent development of purulent vaginal discharge vary with dairy cow breed and parity.

The objective of this study was to characterize the species composition and functional potential of the vaginal and uterine microbiota at 1 wk postpartum in dairy cows diagnosed with or without purulent vaginal discharge (PVD) at 3 wk postpartum. The hypothesis was that differences in the vaginal and uterine microbiota between cows diagnosed with (PVD+) or without (PVD-) PVD were dependent on parity and breed. Cytobrush samples of the vagina and uterus were collected at 1 wk postpartum from 36 Holstein-Friesian (7 primiparous and 29 multiparous) and 29 Jersey (10 primiparous and 19 multiparous) cows. Microbial DNA was isolated from each sample and processed for shotgun metagenomic sequencing. The odds of multiparous cows being diagnosed as PVD+ was less compared with primiparous cows (OR = 0.21). Neither the α-diversity nor ß-diversity of the uterine and vaginal microbiota were associated with PVD but the ß-diversity was different between breeds and between parities. In the vagina of primiparous cows, differences in the microbiota of PVD- and PVD+ cows were minor, but the microbiota of multiparous PVD+ cows had greater relative abundance of Fusobacterium necrophorum, Trueperella pyogenes, Porphyromonas levii, and greater functional potential for amino acid and protein synthesis, energy metabolism, and growth compared with PVD- cows. The uterus of primiparous PVD+ cows had lesser relative abundance of Bacteroides heparinolyticus compared with PVD- cows. In the uterine microbiota, differences included greater functional potential for cellulose biosynthesis and fucose catabolism in multiparous PVD+ cows compared with PVD- cows. In the uterine microbiota of primiparous PVD+ cows, the functional potential for gram-negative cell wall synthesis and for negative regulation of tumor necrosis factor signaling was lesser compared with multiparous PVD+ cows. In the vagina of Holstein-Friesian PVD+ cows, the relative abundance of Caviibacter abscessus was greater whereas in the vagina of Jersey PVD+ cows the relative abundance of Catenibacterium mitsuokai, Finegoldia magna, Klebsiella variicola, and Streptococcus anginosus was greater compared with PVD- cows. In the uterine microbiota of Holstein-Friesian cows, the functional potential for spermidine biosynthesis was reduced compared with PVD- cows. In summary, differences in the species composition and functional potential of the vaginal and uterine microbiota between PVD- and PVD+ cows were dependent on parity and breed. The findings suggest that alternative strategies may be required to treat PVD for different parities and breeds of dairy cow.

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.

Economic impact of different strategies to use sex-sorted sperm for reproductive management in seasonal-calving, pasture-based dairy herds.

To maximize efficiency, profitability, and societal acceptance of modern dairy production, it is important to minimize the production of male dairy calves with poor beef merit. One solution involves using sex-sorted sperm (SS) to generate dairy replacements and breeding all other cows to an easy-calving, short-gestation bull with good beef merit. We used the Pasture Based Herd Dynamic Milk Model to investigate the effect of herd fertility and use of SS on farm net profit in a herd of 100 cows. This was completed by simulating herds with differing fertility performance (good, average, poor), and differing farm reproductive management [conventional semen (CONV) or SS with varying pregnancy per artificial insemination (P/AI) relative to CONV (i.e., relative P/AI 100%, 85%, and 70%)]. As an additional consideration, the method of allocating SS to cows was also examined. The first option used SS on random heifers and cows (S). The second option used SS on heifers and targeted high-fertility cows (SSel). The final option was similar to SSel, but used a fixed-time artificial insemination (AI) protocol to facilitate AI on the farm mating start date (SSync). For CONV, dairy breed semen was used for AI until 50 animals were pregnant (50% chance of a female calf), whereas for S, SSel, or SSync the target number of animals successfully conceiving with SS was set at 28 (based on assumed 90% chance of a female calf from pregnancies derived from SS). Beef breed semen was used on all other dams. The results indicated that the biggest effect on farm net profit was not based on whether or not SS was used, but instead was most affected by the overall fertility performance of the herd. Total farm profit decreased by 10% between the good and average fertility herds, and decreased by a further 12% between the average and poor fertility herds. In almost all situations, when the relative P/AI with SS was ≥85%, use of SS led to an overall increase of the farm net profit. There was an economic benefit of using either SSel or SSync compared with S for the average and poor fertility herds but not for the good fertility herd, highlighting an interaction between SS P/AI and overall herd fertility as well as management practices. If the relative P/AI with SS was <70%, the use of SS led to a decrease in profitability in all simulations except for SSync, highlighting the importance of a good management strategy for use of SS. The findings in this study indicated that SS has significant potential to help facilitate greater integration between the dairy and beef production sectors, as well as increase farm profitability when used appropriately.

Monitoring estrous activity in pasture-based dairy cows.

Correctly identifying cows in estrus and inseminating them at the optimal time are critical components of reproductive management. Technologies for detecting cows in estrus have developed from tail paint in the 1970's to automated activity monitors and mount detectors in recent decades. The objectives of this study were to identify animal characteristics associated with estrous behaviour, measured using a mount detector (FlashMate; Farmshed Labs) and an accelerometer-based activity monitor (Moomonitor, Dairymaster), to examine the relationship between estrous behaviour measured by both devices, and to examine the characteristics associated with pregnancy per AI. Four hundred and sixty eight lactating dairy cows managed on three research farms were enrolled and data were available from 465 cows and 369 cows with Moomonitor and FlashMate data, respectively. Of 234 cows that provided both Moomonitor and FlashMate data, the mean (±SEM) onset of device activation occurred 1.1 (±0.4) h earlier with the Moomonitor compared with the FlashMate. The mean (±SD) duration of Moomonitor activity and FlashMate activity was 17.2 ± 6.1 h and 8.7 ± 5.8 h, respectively. The duration of Moomonitor activity and FlashMate activity was negatively associated with total milk yield during the first five weeks of lactation. The duration of FlashMate activity, but not the duration of Moomonitor activity was positively associated with days in milk. Pregnancy per AI was positively associated with BCS, days in milk, the duration of FlashMate activity and the interval from the onset of Moomonitor and FlashMate activity to AI. Inseminating cows ≤2 h after FlashMate activation or ≤4 h after Moomonitor activation was associated with reduced odds of pregnancy compared with later timing of AI. Overall, 55% of cows received mounts for ≤8 h, highlighting the need for ≥3 periods of estrous observation daily or the use of estrous detection aids that continuously monitor cows. Finally, the study reiterated the importance of maximising body condition score and days in milk at breeding to increase behavioural expression of estrus and pregnancy per AI.

Evaluation of delayed timing of artificial insemination with sex-sorted sperm on pregnancy per artificial insemination in seasonal-calving, pasture-based lactating dairy cows.

The objective was to use ovulation synchronization with timed artificial insemination (TAI) to evaluate the effect of timing of artificial insemination (AI) with frozen sex-sorted sperm on fertility performance in pasture-based compact calving herds. Ejaculates from 3 Holstein-Friesian bulls were split and processed to provide frozen sex-sorted sperm (SS) at 4 × 106 sperm per straw, and frozen conventional sperm at 15 × 106 sperm per straw (CONV). A modified Progesterone-Ovsynch protocol was used for estrous synchronization, with TAI occurring 16 h after the second GnRH injection for cows assigned to CONV, and either 16 h (SS-16) or 22 h (SS-22) for cows assigned to SS. Pregnancy diagnosis was conducted by transrectal ultrasound scanning of the uterus 35 to 40 d after TAI (n = 2,175 records available for analysis). Generalized linear mixed models were used to examine the effects of treatment on pregnancy per artificial insemination (P/AI). Fixed effects included treatment (n = 3), bull (n = 3), treatment by bull interaction, parity (n = 4), days-in-milk category (n = 3), and treatment by days-in-milk category, with herd (n = 24) included as a random effect. Pregnancy per AI was greater for CONV compared with both SS-16 and SS-22 (61.1%, 49.0%, and 51.3%, respectively), and the SS treatments did not differ from each other (relative P/AI for SS-16 and SS-22 vs. CONV were 80.2% and 84.0%, respectively). There were significant bull and treatment by bull interaction effects. Additional analysis was undertaken using a model that included herd as a fixed effect. This analysis identified marked herd-to-herd variation (within-herd relative P/AI for the combined SS treatments vs. CONV ranged from 48-121%). The tertile of herds with the best performance achieved a mean relative P/AI of 100% (range = 91-121%), indicating that P/AI equivalent to CONV is achievable with SS. Conversely, the tertile of herds with the poorest performance achieved a mean relative P/AI of 67% (range = 48-77%). We found that SS resulted in poorer overall P/AI compared with CONV sperm regardless of timing of AI. Marked variation existed between herds; however, one-third of herds achieved P/AI results equal to CONV. Identification of factors responsible for the large herd-to-herd variation in P/AI with SS, and development of strategies to reduce this variation, warrant further research.

Characteristics of offspring derived from conventional and X-sorted bovine sperm.

The objective of this retrospective study was to compare survival during the first year of life and adult performance of offspring derived from artificial insemination (AI) with X-sorted or conventional sperm processed from the same ejaculates. We analyzed a data set that included AI of dairy heifers and lactating cows with fresh conventional sperm (3 × 106 sperm per straw), fresh X-sorted sperm (1 or 2 × 106 sperm per straw), or frozen X-sorted sperm (2 × 106 sperm per straw). The data set contained records of 5,179 offspring born on 396 farms. Offspring were classified as born from conventional sperm (CONV) if they were the product of an insemination with fresh conventional sperm, or born from X-sorted sperm (SS) if they were product of any of the 3 X-sorted sperm treatments. Generalized linear mixed models were used to evaluate the effect of sperm treatment on (1) survival during the first year of life; (2) reproductive performance, lactation performance, and survival of female offspring; and (3) slaughter characteristics of male offspring. Stillbirth rates and mortality rates during the first 2 mo of life were greater for male calves (2.8 and 5.0%, respectively) than for female calves (1.6 and 2.0%, respectively). No differences between offspring derived from SS and CONV were detected for incidences of stillbirth or mortality during the first 12 mo of life within sex of calf. Reproductive performance, milk volume, milk fat, milk protein yields during first; second; and third lactations, and survival to third lactation did not differ between female offspring derived from CONV and SS. Across all age groups, CONV steers had heavier carcasses than SS steers (325.3 vs. 318.3 kg), but there were no differences in weight between CONV and SS steers within any of the age groups (≤24, 25-27, 28-30, and >30 mo of age). The distribution of slaughter age did not differ between CONV and SS steers when the analysis was restricted to herds that reared steers derived from both types of sperm. Carcass conformation and fat scores of steers were not affected by sperm treatment. There was no difference in carcass weight between young bulls (≤2 yr) derived from CONV or SS. In conclusion, the results provide no evidence of differences in survival during the first year of life between offspring derived from CONV or SS, or for any of the reproductive and lactation performance characteristics studied between female offspring derived from CONV or SS. Modest differences in carcass weight between CONV and SS steers were detected, but this may reflect differences in management and husbandry in the rearing herds rather than the sex-sorting process. A controlled study using steers derived from conventional or X-sorted sperm from split ejaculates and reared under the same husbandry conditions is needed to clarify whether there is a true difference in body weight gain due to the sex-sorting process.

Fertility of frozen sex-sorted sperm at 4 × 106 sperm per dose in lactating dairy cows in seasonal-calving pasture-based herds.

The objective was to evaluate the reproductive performance of frozen sex-sorted sperm at 4 × 106 sperm per dose (SexedULTRA 4M, Sexing Technologies, Navasota, TX) relative to frozen conventional sperm in seasonal-calving pasture-based dairy cows. Semen from Holstein-Friesian (n = 8) and Jersey (n = 2) bulls was used. Four of the Holstein bulls used were resident at or near a sex-sorting laboratory (Cogent, UK, or ST Benelux, the Netherlands). The remaining 6 bulls were located at studs in Ireland. For these 6 bulls, ejaculates were collected, diluted with transport medium, and couriered to Cogent in parcel shippers. Transit time from ejaculation to arrival at the sorting laboratory was 6 to 7 h. For all bulls, ejaculates were split and processed to provide frozen conventional sperm (CONV) at 15 × 106 sperm per straw and frozen sex-sorted (SS) sperm at 4 × 106 sperm per straw and used to inseminate lactating dairy cows after spontaneous estrus. Pregnancy diagnosis was performed by ultrasound scanning (n = 7,246 records available for analysis). Generalized linear mixed models were used to examine effects on pregnancy per AI (P/AI) at first artificial insemination, with sperm treatment (CONV vs. SS), bull (n = 10), and treatment × bull interaction as the fixed effects, and herd (n = 142) as a random effect. Overall, P/AI was greater for cows inseminated with CONV than for those inseminated with SS (59.9% vs. 45.5%; 76.0% relative to CONV). This study was not designed to compare resident bulls vs. shipped ejaculates, but the magnitude of the difference between P/AI achieved by CONV and SS was apparently less for resident bulls (60.3% vs. 50.2%) than for shipped ejaculates (58.6% vs. 40.7%). We discovered a treatment × bull interaction for shipped ejaculates (P/AI ranged from 45 to 86% relative to CONV) but not for the resident bulls (P/AI ranged from 81 to 87% relative to CONV). Relative P/AI of SS compared with CONV was greater in cows with high or average fertility potential (76.1% and 78.3%, respectively) than in cows with low fertility potential (58.1%). In 33.1% of the enrolled herds, the P/AI achieved with SS was 90% or more of the P/AI achieved with CONV; this was mainly explained by herds in which SS performed exceptionally well but CONV performed poorly. In conclusion, SS had lower overall P/AI compared with CONV; however, P/AI achieved with SS was dependent on the bull, fertility potential of the cow, and herd. Strategies to improve the P/AI with SS in seasonal-calving pasture-based lactating dairy cows require further research.

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.

Reproductive efficiency and survival of Holstein-Friesian cows of divergent Economic Breeding Index, evaluated under seasonal calving pasture-based management.

The objective of the current study was to examine phenotypic fertility performance and survival, and to gain insight into underlying factors that may contribute to greater fertility performance in 2 divergent genetic groups (GG) of Holstein-Friesian, selected using the Irish Economic Breeding Index (EBI). The GG were evaluated across 3 spring calving pasture-based feeding treatments (FT) over 4 yr. The 2 divergent GG were (1) high EBI; representative of the top 5% nationally (elite), and (2) EBI representative of the national average (NA). In each year, 90 elite and 45 NA cows were randomly allocated to 1 of 3 FT: control, lower grass allowance, and high concentrate. No interaction between GG and FT was observed for any of the measures of fertility investigated. The elite cows achieved significantly greater pregnancy rate to first service (+14.9 percentage points), and significantly greater pregnancy rates after 21, 42, and 84 d of breeding (+17.3, +15.2, and +9.6 percentage points, respectively) compared with NA. The number of services per cow was fewer for elite (1.57) compared with NA (1.80). The interval from mating start date to pregnancy was significantly shorter for elite cows compared with NA. The elite cows maintained greater mean body condition score than NA throughout the study (2.91 vs. 2.72), and had greater body condition score at calving, artificial insemination, and drying off compared with NA. The elite cows had greater mean circulating concentrations of insulin-like growth factor-1 compared with NA. No significant effect was observed of GG on commencement of luteal activity, or progesterone profile variables. Greater survival to the start of fifth lactation was observed for elite cows. The elite cows were 43% less likely to be culled than NA by the beginning of the fifth lactation. The results highlight the success of the Economic Breeding Index to deliver reproductive performance and longevity consistent with industry targets across a range of seasonal pasture-based FT. The results also clearly demonstrate the potential of appropriate genetic selection to reverse negative fertility trends incurred during previous decades of selection for milk production alone.

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.

Genetic merit for fertility traits in Holstein cows: VI. Oocyte developmental competence and embryo development.

The hypothesis of this study was that cows with good genetic merit for fertility traits (Fert+) would produce oocytes and embryos of greater quality than cows with poor genetic merit for fertility traits (Fert-) and that mRNA expression of candidate genes would reflect the observed differences in quality. The aim of the study, therefore, was to determine the effect of genetic merit for fertility traits on morphological classification and mRNA abundance of key genes in immature oocytes and cumulus cells following ovum pick-up and in embryos following superovulation, artificial insemination (AI), and uterine flushing. In experiment 1, 17 Fert+ and 11 Fert- cows, ranging from 54 to 84 d in milk, were submitted to ovum pick-up on 4 occasions during a 2-wk period. Recovered cumulus-oocyte complexes (COC) were morphologically graded. Oocytes and cumulus cells were separated, and mRNA abundance of genes associated with oocyte developmental competence was measured. There was no effect of genotype on the distribution of COC grades or on the mRNA abundance of the candidate genes in grade 1 COC. In experiment 2, 20 Fert+ and 19 Fert- cows, ranging from 71 to 189 d in milk, were submitted to superovulation and AI. The uteri of cows that responded to the superovulation protocol (17 Fert+ and 16 Fert- cows) were nonsurgically flushed 7 d postovulation. Recovered embryos were morphologically graded, and mRNA abundance of genes associated with embryo development was measured in grade 1 blastocysts. The response to the superovulation protocol was assessed by counting the number of codominant follicles on the day of AI, which was similar for both genotypes (22.0 ± 9.7 and 19.8 ± 8.2 for Fert+ and Fert- cows, respectively). There was no effect of genotype on the proportion of transferable embryos recovered or on the mRNA abundance of the candidate genes tested in the grade 1 blastocysts. Of the total embryos classified as blastocysts, however, the Fert+ cows tended to have a greater proportion of grade 1 blastocysts compared with Fert- cows (90% vs. 64%, respectively). In conclusion, genetic merit for fertility traits had a no effect on mRNA abundance of the candidate genes that were examined in immature oocytes and cumulus cells and in embryos recovered after superovulation. The observed differences in morphological blastocyst quality following superovulation would suggest that the superior reproductive performance of Fert+ cows could arise during the later stages of embryo development from d 7 until maternal recognition of pregnancy.

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.