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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, β-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 β-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.

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

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.

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.

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.

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.

peer-reviewed 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.

Long-day photoperiod is known to positively affect milk production in confinement dairy systems, and it has been hypothesized that pineal melatonin (MT) secretion plays a substantial role in this process. Specialized mammalian photoreceptors that regulate MT secretion are optimally stimulated by short wavelength blue light. We investigated the blue light intensity administered to one eye required to suppress MT secretion in nonlactating dairy cows, and subsequently examined effects on milk production in grazing dairy cows. Following a 14-d light-dark 8:16 h environmental conditioning period, 5 nonlactating Holstein-Friesian cows were exposed to treatments of1, 70, 125, 175, and 225 lx for 8 additional hours using a 5 × 5 Latin square design. Light was administered via headpieces fitted with light-emitting diodes emitting blue light (465 nm) to the right eye. All cows were then exposed to a light-dark 16:8 h cycle for one night via the indoor lighting system (200 lx white light). Plasma samples collected at regular intervals were assayed for MT. A dose-dependent effect of light treatment on mean circulating MT concentrations (and 95% CI) was observed [9.4 (7.2, 12.3), 5.0 (3.8, 6.6), 4.4 (3.3, 5.7), 3.3 (2.5, 4.3) and 1.7 (1.3, 2.3) pg/mL for treatments of 0, 70, 125, 175, and 225 lx, respectively. Only the 225 lx treatment acutely suppressed plasma melatonin concentration to levels similar to the light-dark 16:8 h treatment [1.9 (1.4, 2.5) pg/mL]. Forty spring-calving cows were blocked on parity, calving date and Economic Breeding Index for milk production and assigned to the control treatment or blue light to a single eye (LT) treatment from calving through 32 wk of lactation. The cows assigned to LT treatment were fitted with headpieces providing 225 lx of blue light to the right eye from 1700 until 0000 h. Mean milk production (and 95% CI) during 32 wk of lactation was not affected by treatment [20.3 (19.3, 21.3) vs. 20.9 (19.8, 22.0) kg/d, control and LT, respectively]. Within multiparous cows, a treatment by week interaction was detected, whereby LT treatment increased milk production during the first 12 wk of lactation [25.8 (24.3, 27.3) vs. 28.0 (26.5, 29.5) kg/d; +8.5%], but had no effect thereafter. Treatment did not affect plasma insulin-like growth factor 1. We identified the blue light intensity to one eye required to acutely suppress MT concentrations. Transient favorable effects on milk production were observed in multiparous cows. It remains unclear how single-eye blue light treatment affects galactopoiesis in grazing dairy cows, and further research is needed to explore whether this modality of light delivery represents a useful means to aid productivity in pasture-based dairy systems.

The contribution of the calf enterprise to the profit of the dairy farm is generally considered small, with beef bull selection on dairy farms often not considered a high priority. However, this is likely to change in the future as the rapid rate of expansion of the dairy herd in some countries is set to plateau and improvements in dairy herd fertility combine to reduce the proportion of dairy breed calves required on dairy farms. This presents the opportunity to increase the proportion of beef breed calves born, increasing both the value of calf sales and the marketability of the calves. Beef embryos could become a new breeding tool for dairies as producers need to reassess their breeding policy as a consequence of welfare concerns and poor calf prices. Assisted reproductive technologies can contribute to accelerated genetic gain by allowing an increased number of offspring to be produced from genetically elite dams. There are the following 3 general classes of donor females of interest to an integrated dairy-beef system: (1) elite dairy dams, from which oocytes are recovered from live females using ovum pick-up and fertilized in vitro with semen from elite dairy bulls; (2) elite beef dams, where the oocytes are recovered from live females using ovum pick-up and fertilized with semen from elite beef bulls; and (3) commercial beef dams (≥50% beef genetics), where ovaries are collected from the abattoir postslaughter, and oocytes are fertilized with semen from elite beef bulls that are suitable for use on dairy cows (resulting embryo with ≥75% beef genetics). The expected benefits of these collective developments include accelerated genetic gain for milk and beef production in addition to transformation of the dairy herd calf crop to a combination of good genetic merit dairy female calves and premium-quality beef calves. The aim of this review is to describe how these technologies can be harnessed to intensively select for genetic improvement in both dairy breed and beef breed bulls suitable for use in the dairy herd.

peer-reviewed 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 β-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.

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

The objective of this study was to assess the effect of increasing dietary starch for approximately 30 d postpartum on reproduction outcomes in pasture-grazed, seasonal-calving dairy cows. Cows (n = 948) from 3 commercial herds were blocked by age (2, 3, and >3 yr), breed, and expected calving date and randomly assigned to 1 of 2 postpartum treatment groups: high starch (34.7 ± 1.9% nonstructural carbohydrate; mean ± SD) or low starch (22.5 ± 0.4% nonstructural carbohydrate). The high-starch group in all 3 farms received 4.0 to 4.5 kg/d of a 75:25 cracked corn:barley grain mixture in the dairy parlor, split evenly between the morning and afternoon milkings. The low-starch cows received 5.0 to 5.5 kg/d of a 50:50 mixture of palm kernel meal:soy hulls (herds 1 and 3) fed in the parlor; low-starch cows in the remaining herd (herd 2) did not receive a concentrate feed. Cows were cograzed on ryegrass–white clover dominant pastures and were offered corn silage (herds 1 and 3) and canola, corn distillers grain, and palm kernel meal (herd 1) throughout the study. At 1 mo before the start of the seasonal breeding period, the high-starch supplement was removed, and within each herd treatment groups were managed similarly through breeding. Presence of purulent vaginal discharge was assessed at 28 DIM, and tail paint was assessed weekly from 2 to 6 wk postpartum for signs of estrus. The interval to first observed estrus was unaffected by treatment (32.7 vs. 33.5 ± 2 d for high and low starch, respectively), but there were tendencies for a herd × treatment interaction for proportion of cows pregnant to first service and for pregnancy within 6 wk. This interaction was significant for the proportion of cows finally pregnant; a lower proportion of high-starch cows were pregnant to first service, pregnant by 6 wk, and pregnant by the end of the seasonal breeding period in herd 1, but diet did not affect these outcomes in the other herds. Our results do not support a positive effect on reproduction from increasing dietary starch in seasonally bred grazing dairy cows. However, the interactions indicate variability in the herd response to dietary starch early postpartum and imply that pregnancy rate could potentially be compromised through the provision of starch to grazing dairy cows in early lactation (i.e., prebreeding). The experiment was not designed to define the reasons for these interactions, but differences should be considered in future research on the subject.

peer-reviewed The aim of this study was to build and compare predictive models of calving difficulty in dairy heifers and cows for the purpose of decision support and simulation modeling. Models to predict 3 levels of calving difficulty (unassisted, slight assistance, and considerable or veterinary assistance) were created using 4 machine learning techniques: multinomial regression, decision trees, random forests, and neural networks. The data used were sourced from 2,076 calving records in 10 Irish dairy herds. In total, 19.9 and 5.9% of calving events required slight assistance and considerable or veterinary assistance, respectively. Variables related to parity, genetics, BCS, breed, previous calving, and reproductive events and the calf were included in the analysis. Based on a stepwise regression modeling process, the variables included in the models were the dam's direct and maternal calving difficulty predicted transmitting abilities (PTA), BCS at calving, parity; calving assistance or difficulty at the previous calving; proportion of Holstein breed; sire breed; sire direct calving difficulty PTA; twinning; and 2-way interactions between calving BCS and previous calving difficulty and the direct calving difficulty PTA of dam and sire. The models were built using bootstrapping procedures on 70% of the data set. The held-back 30% of the data was used to evaluate the predictive performance of the models in terms of discrimination and calibration. The decision tree and random forest models omitted the effect of twinning and included only subsets of sire breeds. Only multinomial regression and neural networks explicitly included the modeled interactions. Calving BCS, calving difficulty PTA, and previous calving assistance ranked as highly important variables for all 4 models. The area under the receiver operating characteristic curve (ranging from 0.64 to 0.79) indicates that all of the models had good overall discriminatory power. The neural network and multinomial regression models performed best, correctly classifying 75% of calving cases and showing superior calibration, with an average error in predicted probability of 3.7 and 4.5%, respectively. The neural network and multinomial regression models developed are both suitable for use in decision-support and simulation modeling.

From 1917 to 2017, dairy grazing systems have evolved from uncontrolled grazing of unimproved pastures by dual-purpose dairy-beef breeds to an intensive system with a high output per unit of land from a fit-for-purpose cow. The end of World War I signaled significant government investments in agricultural research institutes around the world, which coincided with technological breakthroughs in milk harvesting and a recognition that important traits in both plants and animals could be improved upon relatively rapidly through genetic selection. Uptake of milk recording and herd testing increased rapidly through the 1920s, as did the recognition that pastures that were rested in between grazing events yielded more in a year than those continuously grazed. This, and the invention and refinement of the electric fence, led to the development of "controlled" rotational grazing. This, in itself, facilitated greater stocking rates and a 5 to 10% increase in milk output per hectare but, perhaps more importantly, it allowed a more efficient use of nitrogen fertilizer, further increasing milk output/land area by 20%. Farmer inventions led to the development of the herringbone and rotary milking parlors, which, along with the "unshortable" electric fence and technological breakthroughs in sperm dilution rates, allowed further dairy farm expansion. Simple but effective technological breakthroughs in reproduction ensured that cows were identified in estrus early (a key factor in maintaining the seasonality of milk production) and enabled researchers to quantify the anestrus problem in grazing herds. Genetic improvement of pasture species has lagged its bovine counterpart, but recent developments in multi-trait indices as well as investment in genetic technologies should significantly increase potential milk production per hectare. Decades of research on the use of feeds other than pasture (i.e., supplementary feeds) have provided consistent milk production responses when the reduction in pasture intake associated with the provision of supplementary feed (i.e., substitution rate) is accounted for. A unique feature of grazing systems research over the last 70 yr has been the use of multi-year farm systems experimentation. These studies have allowed the evaluation of strategic changes to a component of the system on all the interacting features of the system. This technique has allowed excellent component research to be "systemized" and is an essential part of the development of the intensive grazing production system that exists today. Future challenges include the provision of skilled labor or specifically designed automation to optimize farm management and both environmental sustainability and animal welfare concerns, particularly relating to the concentration of nitrogen in each urine patch and the associated risk of nitrate leaching, as well as concerns regarding exposure of animals to harsh climatic conditions. These combined challenges could affect farmers' "social license" to farm in the future.

peer-reviewed Anogenital distance (AGD) serves as a marker for prenatal androgenization, reproductive development, and fertility in humans and rodents. The primary objectives of this observational study in lactating dairy cows were to (1) characterize the distribution and variability of AGD, (2) determine the relationship among AGD and potential postnatal AGD determinants of age and height, and (3) evaluate the associations between AGD and pregnancy to first artificial insemination (P/AI) and cumulative pregnancy by 250 d in milk (DIM) within parity groups (first, second, and third+ parities). The secondary objective was to evaluate the association between AGD and testosterone concentrations. The AGD (mm), age (yr), and height at hip (cm) at the time of AGD determination, and aforesaid reproductive outcomes were determined in 921 Holstein cows (first, second, and third+ parity; n = 360, 256, and 305, respectively). Plasma concentrations of testosterone were determined in a subset of 93 cows. Overall, AGD had a normal distribution and high variability [mean (±standard deviation); 131.0 ± 12.2 mm], was weakly associated with cow age and height (coefficient of determination = 0.09 and 0.04, respectively), and had an inverse relationship with P/AI in first- and second-parity cows, but not in third+ parity cows. For every 1 mm increase in AGD, the odds of P/AI decreased by 3.4 and 2.4% for first- and second-parity cows, respectively. The optimal AGD threshold to predict probability of P/AI was 127.1 mm for both first- (sensitivity: 66.4; specificity: 56.6%) and second-parity cows (sensitivity: 46.0; specificity: 70.4%). Accordingly, first- and second-parity cows were categorized into either short or long AGD (≤ or >127.1 mm), and associations with reproductive outcomes were evaluated. First-parity cows with long AGD had lower P/AI (30.9 vs. 53.6%) and decreased likelihood (hazard ratio: 0.68) of pregnancy by 250 DIM than those with short AGD. Similarly, second-parity cows with long AGD had reduced P/AI (28.3 vs. 44.4%) and a tendency for decreased likelihood (hazard ratio: 0.76) of pregnancy by 250 DIM than in cows with short AGD. The association between AGD and testosterone was weak and nonsignificant. In summary, AGD in Holstein cows was normally distributed, highly variable, and weakly associated with age and height. Besides, AGD had an inverse relationship with P/AI and cumulative pregnancy by 250 DIM in first- and second-parity cows; however, such a relationship was not evident in older (third+ parity) cows. This project was financially supported in part by Growing Forward 2, a federal-provincial-territorial initiative (Research Opportunities and Innovation–Internal Initiatives Project # DA7642064). M. Gobikrushanth is a recipient of the Teagasc-University of Alberta Walsh Fellowship sponsored by Alberta Innovates Biosolutions.

peer-reviewed Previous studies have documented that ovarian antral follicle count (AFC) is positively correlated with number of healthy follicles and oocytes in ovaries (ovarian reserve), as well as ovarian function and fertility in cattle. However, environmental factors (e.g., nutrition, steroids) during pregnancy in cattle and sheep can reduce AFC in offspring. The role that genetic and environmental factors play in influencing the variability in AFC and, correspondingly, the size of the ovarian reserve, ovarian function, and fertility, are, however, poorly understood. The present study tests the hypothesis that variability in AFC in offspring is influenced not only by genetic merit but also by the dam age and lactation status (lactating cows vs. nonlactating heifers) and milk production during pregnancy. Antral follicle count was assessed by ultrasonography in 445 Irish Holstein-Friesian dairy cows and 522 US Holstein-Friesian dairy heifers. Heritability estimates for AFC (± standard error) were 0.31 ± 0.14 and 0.25 ± 0.13 in dairy cows and heifers, respectively. Association analysis between both genotypic sire data and phenotypic dam data with AFC in their daughters was performed using regression and generalized linear models. Antral follicle count was negatively associated with genetic merit for milk fat concentration. Also, AFC was greater in offspring of dams that were lactating (n = 255) compared with nonlactating dams (n = 89) during pregnancy and was positively associated with dam milk fat concentration and milk fat-to-protein ratio. In conclusion, AFC in dairy cattle is a moderately heritable genetic trait affected by age or lactation status and milk quality but not by level of dam’s milk production during pregnancy.

A model was developed to examine the effects of sexed semen use in virgin heifers and lactating cows on replacement heifer numbers and rate of herd expansion in a seasonal dairy production system. Five separate herds were established according to the type of semen used: conventional frozen-thawed (Conv), sexed fresh semen used in lactating cows for the first 3 wk of the breeding season (SFre1), sexed frozen-thawed semen used in lactating cows for the first 3 wk of the breeding season (SFro1), sexed fresh semen used in lactating cows for the first 6 wk of the breeding season (SFre2), or sexed frozen-thawed semen used in lactating cows for the first 6 wk of the breeding season (SFro2). In the SFro1, SFre1, SFro2, and SFre2 herds, sexed semen was used for the first and second artificial insemination in virgin heifers. Pregnancy rates achieved with sexed fresh and sexed frozen-thawed semen were assumed to be 94 and 75% of those achieved with conventional frozen-thawed semen, respectively. Initial herd size was 100 cows, which was maintained for the first 2 yr of the 15-yr simulation, after which all available replacement heifers were retained to facilitate herd expansion. Two different scenarios of land availability were examined for each of the 5 herds: land available allowed expansion to a maximum herd size of 150 cows (S1), or land available allowed expansion to a maximum herd size of 300 cows (S2). Once maximum herd size was reached, sexed semen use was discontinued and all excess heifer calves were sold at 1 mo old. All capital expenditure associated with expansion was financed with a 15-yr loan. Each of the 10 different options was evaluated in terms of annual farm profit, annual cash flow, and total discounted net profit. The use of fresh sexed semen generated more replacement heifers, leading to faster herd expansion compared with frozen-thawed sexed semen and conventional frozen-thawed semen. Maximum herd size under S1 was reached in yr 5, 5, 4, 5, and 7 for the SFre1, SFro1, SFre2, SFro2, and Conv herds, respectively. Under S2, maximum herd size was reached in yr 8, 10, 7, 9, and 14 for the SFre1, SFro1, SFre2, SFro2, and Conv herds, respectively. Total discounted net profit under S1 for the SFre2 herd was €4,130, €38,869, €47,231, and €52,089 greater compared with the SFre1, SFro1, Conv, and SFro2 herds, respectively. Under S2, discounted net profit for the SFre2 herd was €44,204, €255,524, €280,373, and €325,815 greater compared with the SFre1, SFro1, SFro2, and Conv herds, respectively. The negative effects of the sexed frozen-thawed semen on herd fertility reduced farm profitability and will prevent its widespread use in lactating cows.

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. Animals60 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, and60 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, and60 DIM, indicating that CIDR-based protocols could be targeted at particular cows, and all other cows could be synchronized using Ovsynch.

A model was developed to examine the effects of sexed semen use on replacement heifer numbers and rate of herd expansion in a seasonal dairy production system. Three separate herds were established according to the type of semen used on virgin heifers: conventional frozen-thawed (Conv), sexed fresh (SFre), or sexed frozen-thawed (SFro). In the model, sexed semen was used for the first and second inseminations in heifers only. Pregnancy rates achieved with sexed fresh and sexed frozen-thawed semen were assumed to be 94% and 75% of those achieved with conventional frozen-thawed semen, respectively. Initial herd size was 100 cows, which was maintained for the first 2 yr of the 15-yr simulation, after which all available replacement heifers were retained to facilitate herd expansion. Two different scenarios of land availability (S1 and S2) were examined for each of the 3 herds using different semen types: land available allowed expansion to a maximum herd size of 150 cows (S1) or 300 cows (S2). Once maximum herd size was reached, sexed semen use was discontinued and all excess heifer calves were sold at 1 mo of age. All capital expenditure associated with expansion was financed with a 15-yr loan. Each of the different options was evaluated in terms of annual farm profit, annual cash flow, and total discounted net profit. The analysis was completed at a milk price of € 0.27/L, and sensitivity around milk price was carried out at € 0.22/L and € 0.32/L. The use of SFre generated more replacement heifers and thus faster herd expansion compared with SFro and Conv semen. Maximum herd size was reached in yr 5, 6, and 7 under S1, and in yr 10, 12, and 14 under S2 for SFre, SFro, and Conv herds, respectively. Total discounted net profit under S1 for the SFre herd was € 19,929 greater than that of the SFro herd and € 41,852 greater than that of the Conv herd. Under S2, discounted net profit for the SFre herd was € 138,587 greater than that of the SFro herd and € 239,987 greater than that of the Conv herd. All 3 herds suffered negative cash flows for extended periods under both S1 and S2 at the lower milk price of € 0.22/L, although cash flows were most negative in the SFre herd. The use of sexed semen, in particular fresh sexed semen, in dairy heifers facilitates faster and more profitable expansion compared with the use of conventional frozen-thawed semen. Financial pressures caused by low milk price were greatest when the rate of expansion was highest.

The primary objective was to determine the variability and repeatability of GnRH-induced LH responses. The secondary objective was to evaluate the associations among plasma LH, FSH, estradiol (E2), and progesterone (P4) concentrations. One hundred lactating Holstein cows (35 primiparous, 65 multiparous) were initially subjected to a presynchronization protocol (d 0, PGF2α; d 3, GnRH) followed 7 d later by Ovsynch (d 10, GnRH; d 17, PGF2α; 56 h later, GnRH) and timed artificial insemination 16 h after the last GnRH. Blood samples were collected immediately before the GnRH injection of presynchronization and the second GnRH of Ovsynch to determine plasma concentrations of LH, FSH, and P4. A second blood sample was collected 2 h after each of the above GnRH injections to determine GnRH-induced LH and FSH concentrations. Plasma concentrations of E2 were also determined in samples collected immediately before the second GnRH of Ovsynch. Cows that (1) had higher LH concentrations at 0 h than at 2 h after GnRH, (2) showed an ongoing spontaneous LH surge, (3) did not respond to GnRH, and (4) had P4 ≥ 0.5 ng/mL at GnRH of presynchronization and the second GnRH of Ovsynch were excluded from the analysis. The variability (coefficient of variation) and repeatability [between animal variance/(within animal variance + between animal variance)] of GnRH-induced LH response were determined from samples collected 2 h after the GnRH of presynchronization and the second GnRH of Ovsynch. The associations among plasma LH, FSH, E2, and P4 were determined at the second GnRH of Ovsynch. Mean (±SEM) LH concentrations before GnRH were 0.5 ± 0.04 and 0.6 ± 0.03 ng/mL, whereas mean LH concentrations 2 h after GnRH were 9.8 ± 1.0 and 12.1 ± 0.8 ng/mL at GnRH of presynchronization and the second GnRH of Ovsynch, respectively. The variability of GnRH-induced LH was 76.1 and 52.1% at GnRH of presynchronization and the second GnRH of Ovsynch, respectively. The repeatability estimate for GnRH-induced LH concentration between GnRH of presynchronization and Ovsynch assessments was 0.10. Plasma concentrations of LH were positively associated with FSH and E2 (r = 0.61 and 0.30, respectively) and negatively associated with P4 (r = -0.46) at the second GnRH of Ovsynch. In summary, GnRH-induced LH responses were highly variable and unrepeatable, and LH concentrations were positively associated with FSH and E2 and negatively associated with P4.

Peer-reviewed This is the author’s version of a work that was accepted for publication in Journal of Dairy Science. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Dairy Science, Volume 95, Issue 7, July 2012, Pages 3711–3721: DOI 10.3168/jds.2011-4977 The objective of this study was to characterize the circulating concentrations of insulin-like growth factor-I (IGF-I) and the hepatic expression of key genes regulating the somatotropic axis in cows divergent in genetic merit for fertility traits but with similar genetic merit for milk production traits. A total of 11 cows with good genetic merit for fertility (Fert+) and 12 cows with poor genetic merit for fertility (Fert−) underwent liver biopsy by percutaneous punch technique on d 20 (± 6.7 d) prepartum and on d 2 (± 1.5 d), d 58 (± 3.7 d), d 145 (± 13 d), and d 245 (± 17.1 d) postpartum. Total RNA was isolated and the mRNA expression of growth hormone receptor (GHR 1A and GHRtot), IGF-I, janus tyrosine kinase 2 (JAK2), signal transducer and activator of transcription 5B (STAT5B), suppressor of cytokine signaling 3 (SOCS-3), acid-labile subunit (ALS), and IGF-binding proteins (IGFBP1 to IGFBP6) were measured by real-time quantitative PCR. During lactation, the circulating concentrations of IGF-I were 34% greater in Fert+ cows. The Fert+ cows had increased mean expression of IGF-I mRNA during the study; however, the difference in IGF-I mRNA abundance between Fert+ and Fert− cows was most pronounced at d 145 and 245. The expression of IGFBP3 and ALS transcript was similar in Fert+ and Fert− cows for the duration of the study. The Fert− cows, however, had greater expression of IGFBP2, IGFBP4, IGFBP5, and IGFBP6. Genotype had no effect on mRNA abundance of GHR 1A, STAT5B, JAK2, or SOCS-3. Genetic merit for fertility traits affects hepatic expression of key genes of the somatotropic axis regulating the synthesis, bioavailability, and stability of circulating IGF-I.

peer-reviewed The total number of ovarian follicles ≥3 mm in diameter (antral follicle count, AFC) during follicular waves varies among cattle of similar age, but AFC is highly repeatable within individuals. We hypothesized that lower AFC could be associated with reduced fertility in cattle. The AFC was assessed by ultrasonography for 2 d consecutively during the first wave of follicular growth of the estrous cycle, 4.6 ± 1.43 d (mean ± SD) after estrus, in 306 Holstein-Friesian dairy cows approximately 70 d postpartum. Cows were classified into 3 groups based on AFC: low (AFC ≤15), intermediate (AFC = 16 to 24), and high (AFC ≥25). During the cycle in which AFC was assessed and in subsequent cycles, cows were artificially inseminated (AI) following detection of estrus, and pregnancy status was assessed using ultrasonography. Cows with high AFC had 3.34 times greater odds of being pregnant at the end of the breeding season compared with cows with low AFC; the odds of a successful pregnancy at first service were 1.75 times greater in the intermediate compared with the low group. The predicted probability of a successful pregnancy by the end of the breeding period (length of breeding season was 86 ± 16.3 d) was 94, 88, and 84% for the high, intermediate, and low AFC groups, respectively. No difference was evident among groups in 21-d submission rate (proportion of all cows detected in estrus and submitted for AI in the first 21 d of the breeding season), but the interval from calving to conception was shorter in the high (109.5 ± 5.1 d) versus low (117.1 ± 4 d) group, and animals with intermediate AFC received fewer services during the breeding season (2.3 ± 0.1) compared with animals with low AFC (2.7 ± 0.1). Lactating cows with ≤15 ovarian follicles have lower reproductive performance compared with cows with higher numbers of follicles, but the existence of a positive association between high numbers of ovarian follicles and fertility is yet to be established.

Peer-reviewed This is the author’s version of a work that was accepted for publication in Journal of Dairy Science. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Dairy Science, Volume 95, Issue 3, July 2012, Pages 1310-1322. DOI 10.3168/jds.2011-4742. The objective of the present study was to characterize the phenotypic performance of cows with similar proportions of Holstein genetics, similar genetic merit for milk production traits, but with good (Fert+) or poor (Fert−) genetic merit for fertility traits. Specifically, we tested the hypothesis that cows with a negative estimated breeding value for calving interval would have superior fertility performance and would have detectable differences in body reserve mobilization and circulating concentrations of metabolic hormones and metabolites compared with cows that had a positive estimated breeding value for calving interval. For the duration of the study, cows were managed identically as a single herd in a typical grass-based, spring-calving production system. A total of 80 lactation records were available from 26 Fert+ and 26 Fert− cows over 2 consecutive years (2008 and 2009). During yr 1, cows were monitored during a 20-wk breeding season to evaluate reproductive performance. Milk production, body condition score (scale 1 to 5), body weight, grass dry matter intake, energy balance, and metabolic hormone and metabolite data were collected during both years. The Fert+ cows had greater daily milk yield (19.5 vs. 18.7 kg/d), shorter interval from calving to conception (85.6 vs. 113.8 d), and fewer services per cow (1.78 vs. 2.83). No difference between groups in grass dry matter intake, energy balance, or body weight was observed. The Fert+ cows maintained greater BCS during mid (2.84 vs. 2.74 units) and late lactation (2.82 vs. 2.73 units). Circulating concentrations of insulin-like growth factor-I were greater throughout the gestation-lactation cycle in Fert+ cows (148.3 vs. 128.2 ng/mL). The Fert+ cows also had greater circulating concentrations of insulin during the first 4 wk of lactation (1.71 vs. 1.24 μIU/mL). Analysis of records from national herd data verified the association between genetic merit for fertility traits and phenotypic reproductive performance; Fert+ cows (n = 2,436) required 11.1 d less to recalve than did Fert− cows (n = 1,388), and the percentage of cows that successfully calved for the second time within 365 and 400 d of the first calving was 8 and 13% greater for Fert+ compared with Fert− cows, respectively. These results demonstrate that genetic merit for fertility traits had a pronounced effect on reproductive efficiency, BCS profiles, and circulating concentrations of insulin-like growth factor-I.

Peer-reviewed Differences in genetic selection criteria for dairy cows internationally have led to divergence in the Holstein-Friesian breed. The objective of this study was to compare hepatic expression of genes of the somatotropic axis in the North American Holstein-Friesian and the New Zealand Holstein-Friesian strains of dairy cow at early and mid lactation. Mature cows of both the North American Holstein-Friesian (n = 10) and New Zealand Holstein-Friesian (n = 10) strains were selected. Liver tissue was collected by percutaneous punch biopsy from all cows at 35 and 140 d postpartum, representing early and mid lactation, respectively. Total RNA was extracted and the hepatic expression of genes involved in the control of the somatotropic axis was examined. Abundance of insulin-like growth factor (IGF)-1 mRNA was greater in the New Zealand strain, concomitant with a tendency for increased expression of acid-labile subunit mRNA. Across strains, mRNA abundance of IGF-binding protein-1, IGF-binding protein-2, and growth hormone receptor 1A decreased from d 35 to 140 postpartum, whereas expression of IGF-1 and acid-labile subunit tended to increase. Abundance of suppressor of cytokine signaling-3 mRNA was increased at d 140 postpartum. Both the strain of Holstein-Friesian cow and the stage of lactation influenced expression of genes controlling the somatotropic axis in hepatic tissue.

Diet-induced milk fat depression in dairy cows has been known for many years and several theories have been proposed. One that continues to receive support is the glucogenic-insulin theory. Previous studies testing this theory using a hyperinsulinemic–euglycemic clamp have had variable results attributable to variability in the use of body fat reserves as a source of milk fatty acids. Our objective was to test the glucogenic-insulin theory using cows immediately postpartum, a period when the use of body fat for milk fat synthesis is greatest. During wk 2 postpartum, 5 cows were given a 2-d baseline period and then clamped for 4 d. Insulin was increased more than 2-fold during the clamp while the blood glucose concentration was maintained. Milk yield was not altered by administration of the clamp (38.7 vs. 39.0±1.4kg/d); however, the milk fat percentage and yield were reduced by 27% and plasma nonesterified fatty acids were reduced by 68%. Analysis of the milk fatty acid composition revealed that the decrease in milk fat yield during use of the clamp was almost exclusively due to reductions in preformed fatty acids; this is the exact opposite of what is observed with diet-induced milk fat depression. Therefore, our results do not support the glucogenic-insulin theory of diet-induced milk fat depression. The results further indicated that reductions in milk fat observed previously with hyperinsulinemic–euglycemic clamps or with glucose or propionate infusions were most likely consequences of the ability of insulin to inhibit lipolysis, thereby limiting the mammary availability of preformed fatty acids mobilized from body reserves.

This study investigated the factors affecting circulating progesterone (P4) concentrations in cows with similar genetic merit for milk production traits, but with extremes of good (Fert+) or poor (Fert-) genetic merit for fertility traits. Study 1: 28 cows were enrolled in an ovulation synchronization protocol at 61±13 (±standard deviation) days postpartum, and data are presented for 13 Fert+ and 9 Fert- cows that remained in the study. Progesterone concentrations were determined from d 0 to 9 (d 0=estrus) and on d 7, corpus luteum (CL) volume and blood flow area (BFA) were measured by B-mode and Doppler ultrasonography, respectively. Cows were administered PGF2α on d 7 in the p.m. and d 8 in the a.m. to regress the CL, and 2 controlled internal drug release devices were inserted per vaginum on d 8 in the a.m. Liver biopsies were collected on d 9 and hepatic mRNA abundance of genes involved in P4 catabolism was determined. On d 10, the controlled internal drug release inserts were removed and frequent blood samples were collected to measure the rate of decline in circulating P4. The Fert+ cows tended to have greater dry matter intake compared with Fert- cows (+0.79kg of dry matter/d), but similar milk production (29.82kg/d). After synchronized ovulation, the rate of increase in circulating P4 concentrations was greater in Fert+ cows compared with Fert- cows. No effect of genotype on CL volume was detected, but BFA was 42% greater in Fert+ cows compared with Fert- cows. The Fert- cows had greater mRNA abundance of cytochrome P450, family 3, subfamily A (CYP3A) compared with Fert+ cows, but the mRNA abundance of aldo-keto reductase family 1, member C1 (AKR1C1), AKR1C3, AKR1C4, and cytochrome P450, family 2, subfamily C (CYP2C) were similar. The half-life and metabolic clearance rate of P4 were similar in Fert+ cows and Fert- cows. Study 2: 23 cows were enrolled in an ovulation synchronization protocol at 55±7 (±standard deviation) d postpartum, and data are presented for 13 Fert+ and 8 Fert- cows that remained in the study. On d 4, 7, 10, and 13 (d 0=estrus), CL volume and BFA were measured as in study 1. Progesterone concentrations were measured from d 1 to 13. Corpus luteum volume was 41% greater in Fert+ cows compared with Fert- cows but no effect of genotype on BFA was detected. Mean circulating P4 concentrations were 79% greater in Fert+ cows compared with Fert- cows. Milk yield was similar in both genotypes. The results indicate that greater circulating P4 concentrations were primarily due to greater CL P4 synthetic capacity rather than differences in P4 clearance in this lactating cow genetic model of fertility.

The aim was to assess the ability of corpus luteum (CL) and uterine ultrasound characteristics on d 18 to 21 to predict pregnancy status in lactating dairy cows. Ultrasound examinations were carried out on cows (n = 164) on d 18 to 21 following artificial insemination (AI). Images of the uterus and CL were captured using a Voluson i ultrasound device (General Electric Healthcare Systems, Vienna, Austria) equipped with a 12-MHz, multi frequency, linear array probe. Serum concentrations of progesterone were determined from blood samples collected at each ultrasound examination. Images of the CL were captured and stored for calculation of CL tissue area and echotexture. Images of the CL and associated blood flow area were captured and stored for analysis of luteal blood flow ratio. Longitudinal B-mode images of the uterine horns were stored for analysis of echotexture. Diagnosis of pregnancy was made at each ultrasound examination based on CL blood flow, CL size, and uterine echotexture. Pregnancy was confirmed by ultrasonography on d 30 after AI. The relationship between ultrasound measures and pregnancy outcome, as well as the accuracy of the pregnancy diagnosis made at each ultrasound examination was assessed. Progesterone concentrations and CL tissue area were greater in pregnant compared with nonpregnant cows on all days. The CL blood flow ratio was higher in pregnant compared with nonpregnant cows on d 20 and 21 after AI. Echotexture measures of the CL and uterus were not different between pregnant and nonpregnant cows on any day of examination. The best logistic regression model to predict pregnancy included scores for CL blood flow, CL size, and uterine echotexture on d 21 following AI. Accuracy of pregnancy diagnosis was highest on d 21, with sensitivity and specificity being 97.6 and 97.5%, respectively. Uterine echotexture scores were similar for pregnant and nonpregnant cows from d 18 to 20. On d 21, pregnant cows had higher uterine echotexture scores compared with nonpregnant cows. The logistic regression equation most likely to provide a correct pregnancy diagnosis in lactating dairy cows included the visual score for CL blood flow, CL size, and uterine echotexture on d 21 after AI. In support of this finding, the diagnostic accuracy for visual scores of CL blood flow, CL size, and uterine echotexture were also highest on d 21.

The objective of this study was to monitor the dry matter intake (DMI), metabolic status, uterine health, and resumption of cyclicity in cows with similar genetic merit for milk production traits but with either good (Fert+) or poor genetic merit (Fert-) for fertility traits. Twenty-six cows were enrolled in the study and data are reported for 15 Fert+ and 10 Fert- cows that completed the study. All cows received a total mixed ration diet during early lactation and were turned out to pasture in late spring. Dry matter intake was recorded daily from wk -2 to 5 relative to parturition. Blood metabolites and metabolic hormones were measured from wk -2 to 8 relative to parturition. Milk production, body condition score, and body weight until wk 35 of lactation are reported. To monitor uterine health, vaginal mucus was scored weekly on a scale of 0 (no pus) to 3 (≥ 50% pus) from parturition to wk 8 and uterine polymorphonuclear neutrophil count was measured at wk 3 and 6 postpartum. Prepartum DMI was similar between genotypes, but Fert+ cows had significantly greater DMI than Fert- cows (19.7 vs. 16.8 kg of dry matter/d) during the postpartum period. Energy balance at wk 1 was significantly greater in Fert+ cows than in Fert- cows [2.3 vs. -1.12 unite fourragere lait (UFL)/d]. The Fert+ cows had significantly greater daily milk solids production (1.89 vs. 1.74 kg/d) and tended to have greater daily milk yield (24.2 vs. 22.3 kg/d). The Fert+ cows had significantly greater mean circulating insulin-like growth factor-I (102.62 vs. 56.85 ng/mL) and tended to have greater mean circulating insulin (3.25 vs. 2.62 μIU/mL) compared with Fert- cows from wk -2 to 8 relative to parturition. Mean circulating glucose (3.40 vs. 3.01 mmol/L) concentrations were significantly greater in Fert+ cows compared with Fert- cows from wk -2 to 3 relative to parturition. The Fert+ cows maintained significantly greater mean body condition score throughout lactation compared with Fert- cows (2.98 vs. 2.74 units). Moreover, Fert+ cows had better uterine health compared with Fert- cows, as evidenced by lower weekly vaginal mucus scores from wk 2 to 6 postpartum and, based on uterine cytology, smaller proportions were classified as having endometritis at wk 3 (0.42 vs. 0.78) and 6 (0.25 vs. 0.75). Also, a significantly greater proportion of Fert+ cows had resumed cyclicity by wk 6 postpartum (0.86 vs. 0.20) compared with Fert- cows. Hence, we report for the first time that genetic merit for fertility traits is associated with postpartum uterine health status. Superior uterine health and earlier resumption of cyclicity may be mediated through differences in DMI, energy balance, insulin, insulin-like growth factor-I, and body condition score profiles. Importantly, phenotypic improvement in fertility traits was achieved without antagonizing milk production.

The objective of this study was to compare circulating progesterone (P4) profiles and pregnancies per AT (P/AI) in lactating dairy cows bred by timed artificial insemination (TAT) 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(2 alpha)-3d-GnRH-7d-Ovsynch-56 (GnRH-7d-PGF(2 alpha)-56h-GnRH-16hTAI)] or PS (n = 850; PGF(2 alpha)-14d-PGF(2 alpha)-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 74 d, and data were available on P/AI at the subsequent second service. Presynchronization with DO reduced the percentage of animals with low P4 concentrations (

Peer-reviewed This is the author’s version of a work that was accepted for publication in Journal of Dairy Science. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Dairy Science, volume 95 (7), July 2012 The objective of this study was to characterize the estrous cycle of cows with similar proportions of Holstein genetics, similar genetic merit for milk production traits, but with good (Fert+) or poor (Fert-) genetic merit for fertility traits. A total of 37 lactating cows were enrolled on an 8-d CIDR-based protocol to synchronise estrus. 19 Fert+ and 12 Fert- cows that successfully ovulated a dominant follicle and established a corpus luteum underwent daily transrectal ultrasonography. Blood sampling was carried at 8 h intervals from d 0 to d 6 and from d 15 to ovulation, and once daily from d 7 to d 15. Blood samples were analysed for progesterone, estradiol, follicle stimulating hormone and luteinising hormone. Estrus behaviour was recorded using neck activity collars and mounting pads. Fert+ cows tended to have fewer (P = 0.07) follicular waves (2.2 vs. 2.7 waves) and had a shorter (P < 0.05) estrous cycle (21.0 vs. 25.1 d) than Fert- cows. There was no effect of genotype on day of first wave emergence or day of first wave dominant follicle peak diameter (all P > 0.05) but the peak diameter of the first wave dominant follicle tended to be larger (P = 0.08) in Fert- cows. During the first 13 d of the cycle, Fert+ cows developed a corpus luteum that was 16% larger (P = 0.08) than Fert- cows. Circulating progesterone concentrations were 34% greater (P < 0.001) in Fert+ than Fert- cows (5.15 vs. 3.84 ng/ml, respectively) from d 5 to d 13. During the final follicular wave, the interval from preovulatory follicle emergence to ovulation and the interval from preovulatory follicle dominance to ovulation were similar (P >0.05) in both genotypes. Maximum preovulatory follicle diameter was larger (P < 0.05) in Fert+ than Fert- cows (17.9 vs. 16.8 mm, respectively); however, circulating concentrations of oestradiol were not different (all P > 0.05) between genotypes. A greater proportion (P < 0.05) of Fert- cows ovulated to a silent heat than Fert+ cows (22% vs. 2%, respectively). Of cows that showed behavioural estrus Fert+ cows had 41% greater (P < 0.01) mean activity count; however, no difference (P > 0.05) was seen in mounting behaviour between genotypes. These results demonstrate for the first time that genetic merit for fertility has pronounced effects on corpus luteum development, progesterone concentration, preovulatory follicle diameter and behavioural estrus.

Peer-reviewed This is the author’s version of a work that was accepted for publication in Journal of Dairy Science. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Dairy Science, Volume 95, Issue 5, May 2012, Pages 2442-2451: DOI 10.3168/jds.2011-4632 The objective was to determine the effects of a protected (lipid-encapsulated) conjugated linoleic acid (LE-CLA) supplement on milk production, estrous cycle characteristics, and reproductive performance in lactating dairy cows on a pasture-based diet. Spring calving dairy cows (n = 409) on a single pasture-based commercial dairy farm were used in a completely randomized block design. Cows were assigned to 1 of 2 dietary supplements [LE-CLA (n = 203) or no supplement (control, n = 206)]. The LE-CLA cows received 51 g/d of a lipid supplement containing 5 g of both trans-10,cis-12 and cis-9,trans-11 CLA from 0 to 60 d in milk. Milk samples were collected 3 times weekly, and each sample was analyzed for progesterone to determine the interval to first ovulation and estrous cycle characteristics. Milk yield and concentrations of fat, protein, and lactose were measured every 2 wk. Cows were inseminated following visual observation of estrus. The breeding season commenced on April 8, 2009 and continued for 16 wk. Transrectal ultrasonography was carried out at 30 to 36 d and 60 to 66 d post-AI to diagnose pregnancy. The LE-CLA treatment resulted in a decrease in milk fat concentration (36.9 ± 0.06 g/kg vs. 30.7 ± 0.06 g/kg for control and LE-CLA, respectively) and yield (0.91 ± 0.02 kg/d vs. 0.84 ± 0.02 kg/d for control and LE-CLA, respectively); however, milk yield was increased by LE-CLA supplementation (24.7 ± 0.7 kg/d vs. 27.2 ± 0.7 kg/d for control and LE-CLA, respectively), resulting in no overall difference in milk energy output. No effect of LE-CLA was observed on any estrous cycle characteristics or measures of reproductive performance. These results support that in pasture-based systems of dairy production, where energy intake limits milk production, energy spared by CLA-induced milk fat depression is partitioned toward increasing milk yield rather than toward body reserves.

This study was carried out to examine the effects of decreasing the dry period (DP) duration and altering the energy density of the diet during early lactation on the rheological characteristics of milk. Forty mature Holstein-Friesian cows were used in a completely randomized design with a 2 x 2 factorial arrangement of treatments. Cows were randomly assigned to 1 of 2 dry period treatments and 1 of 2 nutritional treatments. The DP treatments were continuous milking (CM) or an 8-wk standard dry period (SDP), and the nutritional treatments were a standard energy diet (SE) or a high energy diet (HE). Actual DP lengths were 6.3+/-1.7 d and 62.1+/-1.9 d for cows on the CM and SDP treatments, respectively. Milk samples were collected at 2, 6, and 10 wk postpartum. The concentrations of fat, protein, and lactose were determined in each sample. The rennet gelation properties were measured at 31 degrees C using dynamic low-amplitude strain oscillatory rheometry. The following parameters were obtained from the resultant elastic shear modulus (G'): gelation time, maximum curd firming rate, and gel strength. Reducing the DP duration from 62 to 6 d resulted in increases in milk protein concentration (31.8 vs. 34.7 g/kg), maximum curd firming rate (2.58 vs. 3.60 Pa/min), and gel strength (69.4 vs. 90.5Pa). Increasing the dietary energy density decreased percentage milk fat (43.1 vs. 37.7 g/kg) but otherwise had no effect. Gel strength was correlated with maximum curd firming rate (r=0.99), and both variables were correlated with milk protein concentration (r=0.71 and r=0.73, respectively). The results indicate that decreasing the duration of the DP increased milk protein concentration and improved the rennet gelation properties of milk, but that dietary energy density had little effect.

High proportions of embryonic and early fetal losses in dairy cattle are associated with low peripheral concentrations of progesterone, which could result from increased catabolism, decreased production, or both. Progesterone catabolism occurs primarily in the liver via the cytochrome P450 2C (CYP2C) and cytochrome P450 3A (CYP3A) subfamilies (EC 1.14.14.1; unspecific monooxygenases). Recent observations from our laboratory have shown that the fractional rate constant of progesterone decay can be dramatically reduced by insulin because of a decrease in hepatic CYP2C and CYP3A activity. Little information exists on the regulation of progesterone catabolic enzymes in dairy cows. We hypothesized that elevated insulin concentrations would down-regulate hepatic CYP2C and CYP3A mRNA; therefore, our objectives were to determine the relative abundance of hepatic CYP2C and CYP3A mRNA in dairy cows in response to elevated concentrations of insulin. In the first experiment, 17 mature Holstein cows were drenched daily with 500 mL of water (n = 10) or propylene glycol (a gluconeogenic substrate; n = 7) from 10 d before their expected calving date until d 25 postpartum. Cows drenched with propylene glycol had a 30% increase in peripheral concentrations of insulin. Liver biopsies were collected on d 25 postpartum to determine the relative abundance of CYP2C and CYP3A mRNA. In the second experiment, 19 mature, lactating Holstein cows were randomly assigned to a hyperinsulinemic-euglycemic clamp (0.3 or 1.0 microg of insulin/kg of BW per h; n = 6 each) or remained as controls (saline infused; n = 7) for 96 h beginning on d 10 postpartum. Insulin infusion resulted in a 2.6- or 8- fold increase in peripheral concentrations of insulin, respectively. On d 14 postpartum, a liver biopsy was collected to determine CYP2C and CYP3A mRNA abundance. In experiment 1, the relative abundance of CYP2C mRNA in cows treated with propylene glycol did not differ from controls; however, the relative abundance of CYP3A mRNA in the propylene glycol group was 63% that of controls. For experiment 2, there was a dose-dependent decrease in the relative abundance of both CYP2C and CYP3A mRNA with increasing dosage of insulin. In conclusion, this study demonstrated that, in the cow, either providing a gluconeogenic feed-stuff or treatment with insulin decreased the abundance of mRNA for enzymes responsible for hepatic progesterone catabolism.

There is a paucity of studies on the effect of intrauterine conditions on subsequent progeny performance in dairy cows. Using a large national data set on Irish Holstein-Friesian dairy cows, the objective of this study was to determine if intrauterine conditions, quantified by a maternal genetic variance component, significantly affected milk production, age at first calving, calving interval, somatic cell score (natural logarithm of somatic cell count) and survival in first-, second-, and third-parity female offspring. Maternal genetic variance for each trait in each parity was estimated in a linear mixed model which included, other than fixed effects, direct additive genetic, maternal genetic, cytoplasmic and permanent environmental effect of the dam, and residual component. A covariance was also estimated between the direct additive and maternal genetic components where possible. Because calves in Irish dairy herds are separated from dams at birth, a significant maternal genetic variance (with all other random effects in the model) indicates a prepartum maternal effect. A significant maternal genetic variance was estimated for 305-d milk yield in first and third lactation, somatic cell score in first lactation, and survival to second lactation from 188,144 lactations on 80,881 animals. Where estimated, a negative correlation existed between the direct additive and maternal genetic components. Regression of maternal mixed model solutions on dam milk production at different stages relative to conception revealed that greater milk yield preconception and during gestation was associated with reduced survival and milk yield and greater somatic cell count in the progeny. This study suggests that offspring survival and performance are affected by prepartum conditions that offspring experience as an oocyte, embryo, or fetus, one of which is mediated through milk production (or factors related to milk production) of the dam.

The present study was carried out to 1) evaluate the viability of in vitro fertilized zygotes after microinjection of DNA, 2) assess the influence of oocyte quality upon the development rate of embryos when injected with DNA, and 3) determine the integration frequency of green fluorescent protein DNA into microinjected embryos. Oocytes were aspirated from ovaries of nine nonlactating Holsteins and were categorized into grades A, B, C, and D. At 16 h after in vitro fertilization, approximately half of the pronuclear stage presumptive zygotes were classified as having 1 pronucleus or 2 pronuclei, and they were microinjected with DNA constructs. A potential predictor of DNA integration frequency at d 10 was assessment of the incidence of green fluorescing embryos. The proportion of cleaved embryos that developed to morulae or blastocysts was not different between groups with 1 pronucleus injected (45%), 1 pronucleus uninjected (64%), or 2 pronuclei injected (49%). However, the development of morulae or blastocysts was higher in the group with 2 pronuclei uninjected (69%). The overall developmental score of green fluorescent protein-positive embryos was higher for grade A oocytes (1.3 +/- 0.1) than for grade B (0.8 +/- 0.1), C (0.6 +/- 0.1), or D (0.3 +/- 0.1) oocytes. The results show that production of transgenic bovine blastocysts can occur from the microinjection of a presumptive zygote having only one visible pronucleus. Initial oocyte quality is an important factor in selection of oocytes suitable for microinjection of DNA and for preimplantation development to produce bovine transgenic embryos.