B vitamins, including niacin (vitamin B3), are synthesized by rumen microbes, but supplementation may provide additional benefits for ruminant health and productivity. Supplementing rumen-protected niacin (RPN) during the transition period can reduce lipolysis after calving and, consequently, may improve health and fertility of dairy cattle later in lactation. Our objective was to determine if supplementing RPN during the first 21 DIM would improve the health of dairy cows on a commercial dairy farm. We hypothesized that RPN would reduce mastitis, improve fertility, and reduce risk of leaving the herd during lactation. Holstein cows were blocked by parity and projected calving date, with treatments randomly assigned to cows within each block. Cows received RPN (n = 481; 26 g/d) through a supplement dispenser in the automated milk systems (AMS) in addition to their robot pellet, or the robot pellet only (CON; n = 593). Treatments were applied for the first 21 DIM, and cows were followed for the rest of their lactation. Milk yield, milk components (wk 1-3 of lactation), pre- and postpartum BCS, health records, and reproductive records were collected. Blood was collected from a random subset of 99 cows at 3 DIM and 97 cows at 10 DIM to assess plasma concentrations of niacin, metabolic biomarkers, and biomarkers of inflammatory status. Culling, proportion of cows pregnant, and mastitis risk were analyzed using Cox proportional hazard models. Mastitis incidence was analyzed with a linear mixed model and conception risk was analyzed using a chi 2 test. Supplementing RPN increased plasma nicotinamide concentration by 1,740 nM +/- 410.0 nM (SE; 50% increase), but it did not affect plasma nicotinic acid concentrations. Supplementing RPN reduced plasma insulin concentrations at 3 and 10 DIM across all parities. Circulating BHB and free fatty acid concentrations were greater for cows receiving RPN; the effect was greatest in cows in third or greater parity. Plasma haptoglobin was not affected by treatment. Rumen-protected niacin increased milk yield for first- and second-parity cows by wk 9 and 13 of lactation, respectively, and increased milk yield in those groups was sustained for the rest of lactation, resulting in 658 +/- 259.4 kg and 675 +/- 308.9 kg more milk for RPN-supplemented first- and second-parity cows. The risk of leaving the herd, mastitis incidence, and probability of pregnancy were not affected by RPN. Inquiries into the effects of RPN supplementation and its timing on metabolism are necessary to understand optimal supplementation strategies for RPN in dairy cattle.

The objective of this study was to determine the effect of the timing of presumptive conceptus attachment (pCA) and its relationship with subsequent pregnancy loss (PL) in lactating dairy cows following timed artificial insemination with conventional (TAI-C) or X-sorted (TAI-S) semen or timed embryo transfer (TET) with frozen-thawed in vitro-produced (IVP) embryos. Lactating cows (n = 417, predominantly Holstein-Friesian) located in 3 herds were blocked based on parity, calving date, and economic breeding index and randomly assigned to receive TAI-C (n = 136), TAI-S (n = 136) or TET with a single, grade-1 frozen-thawed IVP blastocyst (n = 145). Cows were synchronized with a 10-d Progesterone-Ovsynch protocol, and were either inseminated (TAI-C, TAI-S) 16 h after the second GnRH or received TET on d 7 after presumptive estrus. Serum progesterone was measured in all cows on d 7 after expected ovulation. For all cows that had not returned to estrus, serum pregnancy-specific protein B (PSPB) was measured on d 7, 17, and daily from d 19 through 28 after expected ovulation to characterize the timing of pCA. The day of pCA was defined as the first day of an increase in PSPB of ≥12.5% from baseline (d 17) followed by 2 more consecutive days of ≥12.5% increase from the previous day. Pregnancy was diagnosed in cows that had not returned to estrus via ultrasound examination on d 32, 62, and 120 postovulation, and calving data were recorded. Day of pCA (mean; 95% CI) was earlier for TAI-C (20.0; 19.7, 20.3) compared with TET (20.6; 20.3, 20.9), and TAI-S (20.3; 19.9, 20.6) was not different from the other 2 treatments. Calving/service event (C/SE) was greater (83.2% vs. 54.4%) and PL during the interval from pCA to expected calving date was less (16.8% vs. 45.6%) for cows with early pCA (≤d 20; 23/137) compared with cows that had late pCA (≥d 21; 36/79). The incidence of PL was greater for cows assigned to TAI-S (30.7%) and TET (33.8%) than TAI-C (16.4%). Consistent with previous studies, measuring time to pCA provides new insights into understanding early pregnancy establishment and PL in lactating dairy cows. The timing of pCA was later for TET compared with TAI-C and may be associated with increased risk of PL. Further research into the regulatory mechanisms underlying conceptus attachment is warranted.

Prediction of pregnancy survival in lactating dairy cows can be determined by the conceptus attachment timeframe via daily pregnancy-specific protein B (PSPB) monitoring. All factors contributing to reduced fertility in dairy cows receiving AI following estrus detection remain unclear. This study aimed to determine differences in time to conceptus attachment in lactating cows treated with the fertility program Double-Ovsynch compared to cows that were detected in estrus. Additionally, we investigated various pre- and post-conception factors potentially influencing fertility outcomes. We hypothesized that AI following a natural estrus detected with automated activity monitors would lead to an extended time to conceptus attachment and lower PSPB concentrations post-attachment compared to Double-Ovsynch. There were no differences in the average time to conceptus attachments between treatments. However, cows inseminated post-estrus that experienced pregnancy loss between conceptus attachment and 60-66 days post-AI exhibited diminished PSPB concentrations on Days 2 and 3 following conceptus attachment. Steroid hormone interactions were assessed with radioimmunoassay to determine the ratios of estrogen to progesterone concentrations on the day of the luteinizing hormone (LH) surge. Notably, estrogen to progesterone ratio proved to predict conceptus attachment in cows subjected to Double-Ovsynch but not in those inseminated post-estrus detection surge. In conclusion, the estrogen to progesterone ratio measured around the time of the pre-ovulatory LH surge emerges as a potentially effective tool for estimating the fertility potential of lactating dairy cows undergoing timed AI, particularly in the context of the Double-Ovsynch program.

Cloning cattle using somatic cell nuclear transfer (SCNT) is inefficient. Although the rate of development of SCNT embryos in vitro is similar to that of fertilized embryos, most fail to develop into healthy calves. In this study, we aimed to identify developmentally competent embryos according to blastocyst cell composition and perform transcriptome analysis of single embryos. Transgenic SCNT embryos expressing nuclear-localized HcRed gene at day 7 of development were imaged by confocal microscopy for cell counting and individually transferred to recipient heifers. Pregnancy rates were determined by ultrasonography. Embryos capable of establishing pregnancy by day 35 had an average of 117 & PLUSMN; 6 total cells, whereas embryos with an average of 128 & PLUSMN; 5 cells did not establish pregnancy (P < 0.05). A lesser average number of 41 & PLUSMN; 3 cells in the inner cell mass (ICM) also resulted in pregnancies (<0.05) than a greater number of 48 & PLUSMN; 2 cells in the ICM. Single embryos were then subjected to RNA sequencing for transcriptome analysis. Using weighted gene coexpression network analysis, we identified clusters of genes in which gene expression correlated with the number of total cells or ICM cells. Gene ontology analysis of these clusters revealed enriched biological processes in coenzyme metabolic process, intracellular signaling cascade, and glucose catabolic process, among others. We concluded that SCNT embryos with fewer total and ICM cell numbers resulted in greater pregnancy establishment rates and that these differences are reflected in the transcriptome of such embryos.

The development of fertility programs and their adoption by the dairy industry over the past decade is a major driving factor underlying the dramatic increase in reproductive performance in lactating dairy cows during the past 20 years. Another major driving factor underlying this increase in reproductive performance is what we describe in this minireview as the "high fertility cycle." We now know that reproductive performance and the incidence of certain periparturient health events are interrelated. The high fertility cycle describes the relationship between body condition score (BCS) change during the periparturient period and postpartum health events and subsequent reproductive performance in which lactating dairy cows that establish pregnancy by 130 d in milk have shorter calving intervals and thereby gain less BCS during the current lactation and dry off and calve at a lower BCS (2.75 to 3.0) than cows with a longer lactation. After calving, these cows undergo less BCS loss, experience fewer health issues, have greater fertility at first insemination, and have reduced early pregnancy losses after establishment of pregnancy and thereby become pregnant before 130 d in milk. This minireview overviews these relationships and highlights the key concepts underlying the high fertility cycle. Future randomized, controlled experiments are needed to causally link these relationships between BCS change and fertility in lactating dairy cows.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Increasing progesterone (P4) during early conceptus development may be crucial for establishment of pregnancy in dairy cattle. The objective of this study was to determine if human chorionic gonadotropin (hCG) at various times after ovulation will increase serum P4 during elongation and increase the chances for, and reduce variability to, initial increase in pregnancy-specific protein B (PSPB) following artificial insemination (AI). Time to PSPB increase was defined as the first day of increase in concentrations of PSPB between d 18 and 28 after ovulation in cows with ≥12.5% increases for 3 consecutive days compared with baseline. Lactating cows (n = 368) synchronized to Double-Ovsynch (first service) or Ovsynch (second or greater service) received one of 4 treatments: no hCG (control), or 3,000 IU of hCG on d 2 (D2), 2 and 5 (D2+5), or 5 (D5) after ovulation. All cows were examined via ultrasound on d 5 and 10 postovulation to determine percentage of cows with hCG-induced accessory CL (aCL) and to quantify and measure all luteal structures. Samples for serum P4 were collected on d 0, 5, 19, and 20 postovulation. The P4 was increased in D2, D2+5, and D5 groups compared with control. The D2+5 and D5 treatments increased aCL and P4 compared with D2 and control. The D2 treatment increased P4 on d 5 after ovulation compared with control. Serum PSPB samples were collected daily from all cows on d 18 through 28 after ovulation for determination of d of PSPB increase. Pregnancy diagnoses were performed via ultrasound examination on d 35, 63, and 100 after ovulation and AI. The D5 treatment reduced percentage of cows with, and increased the time to, PSPB increase. Primiparous cows with ipsilateral aCL had reduced pregnancy loss before d 100 postovulation compared with cows with contralateral aCL. Cows that had PSPB increase >21 d postovulation had 4× greater chances of pregnancy loss compared with cows that had PSPB increase on d 20 or 21. The highest quartile of P4 on d 5, but not on d 19 and 20, was associated with reduced time to PSPB increase. Time to PSPB increase appears to be an important measurement to understand reasons for pregnancy loss in lactating dairy cows. Increasing P4 utilizing hCG after ovulation did not enhance early pregnancy or reduce pregnancy losses in lactating dairy cows.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

Lactating dairy cows have reduced chances for pregnancy survival following 1st parturition. It appears that timing of conceptus attachment, defined as an initial daily rise in pregnancy-specific protein B (PSPB) postartificial insemination (AI), is key to the survival of the embryo. PSPB is a protein produced from binucleate cells of the trophectoderm of the conceptus. Once the conceptus has attached to the uterine epithelial cells, this large protein is detectable in maternal circulation. Daily sampling of serum allows for detection of the initial increase of PSPB. A percent increase threshold for PSPB was determined from results from each study using a conservative three-day minimum increase. Thus far, a 10 or 12.5% daily increase for three consecutive days had the greatest sensitivities and specificities for studies reported in this review. Nulliparous heifers receiving AI following estrus appear to have a reduced time to conceptus attachment compared to lactating cows that received either Ovsynch or AI following estrus. Increasing progesterone post-AI did not reduce time to conceptus attachment in lactating cows. Multiparous cows treated post-AI with human chorionic gonadotropin had a reduced percent with conceptus attachment in addition to greater time to conceptus attachment. Increased time to conceptus attachment increased the chances of multiparous cows having embryonic death before 35 d post-AI. Concentrations of PSPB are consistently reduced from conceptus attachment until d 28 post-AI in cows with pregnancy loss compared to cows that maintain pregnancy on d 34 post-AI. This novel model to determine timing of conceptus attachment provides a new perspective on percent of cows pregnant around d 20-23 post-AI and the potential for a greater understanding of subsequent pregnancy loss.

The onset of productive life in dairy cattle, concomitant to parturition, is accompanied by a substantial decrease in fertility in comparison with non-lactating, nulliparous heifers. Follicular growth patterns differ between parous and nulliparous dairy cattle. Nulliparous heifers ovulate follicles with reduced antral age (RAA). This study aimed to exogenously reduce ovulatory follicle age in lactating dairy cows from 7 to 5 days old. Cows (n = 80) had their estrous cycles synchronized with the Double-Ovsynch program. At the final portion of this program, luteolysis was induced at either 5 (RAA) or 7 (Control) days following follicular wave emergence. RAA outcomes were estimated in comparison with Controls. RAA resulted in smaller follicles 2 days post-treatment. Despite lower serum concentrations of 17β-estradiol before treatment compared with Controls, the rate of increase in this hormone was greater for the RAA treatment. There was no difference in luteolysis rates between treatments. Proestrus (luteolysis onset to estrus onset) was prolonged in RAA cows. Cows with RAA had more intense estruses. Collectively, these results indicate that decreasing the age of the ovulatory follicle may improve the steroidogenic capacity of the dominant follicle and estrus expression intensity in lactating dairy cows.

Reproduction, Fertility and Development is an international journal publishing original research , review and comment in the fields of reproduction and developmental biology in humans, domestic animals and wildlife

This study was designed to use within-individual daily increases in circulating pregnancy-specific protein B (PSPB) to determine time to increase in serum PSPB in nulliparous heifers and multiparous dairy cows following artificial insemination (AI). Weekly cohorts of lactating Holstein cows (n = 56) averaging 122 ± 7 d in milk at AI and nulliparous Holstein heifers (n = 23) averaging 16 ± 0.24 mo old at AI were used in this study. Lactating cows received AI to either Ovsynch (n = 38) or at ~12 h following observed standing estrus (n = 18). All heifers were inseminated ~12 h after observed standing estrus. Blood samples for measurement of PSPB were collected daily from d 15 through d 35 postovulation. Nulliparous heifers had increases in serum PSPB earlier compared with both primiparous and multiparous cows. Day of increase in serum PSPB was defined as the day serum PSPB optical density levels initially increased ≥10% from baseline and continued to increase from baseline of ≥10% the following 2 d. Average PSPB were greater in pregnant heifers compared pregnant cows from d 23 through 29. Early lactation nonpregnant cows maintained greater average optical density of serum PSPB from 15 to 35 d postovulation compared with nonpregnant heifers (n = 38). In summary, fertility differences in heifers versus lactating cows may be due to the differences in timing of increases in serum PSPB. This appears to be one of the first publications that used daily PSPB sampling to investigate possible differences in fertility in heifers versus lactating cows.

The objective of this study was to determine the effect of early lactation body condition (BC) loss in multiparous dairy cows on serum lipids and the effect of these changes on oocyte and cumulus cell transcriptomes. Body condition loss in dairy cattle after parturition is associated with reduced fertility and increased pregnancy loss. The complex interplay between BC, nutrition, dry matter intake, milk production, and time of calving has presented a barrier to understanding mechanisms leading to reduced fertility. We identified cows that lost BC (L group; n = 10) or maintained or gained BC (M/G group; n = 8) during the first 27 to 33 d in milk and investigated changes in serum fatty acids and oocyte and cumulus cell transcriptomes at 75 to 81 d in milk. The L group had increased serum levels of nonesterified fatty acids and mead acid, and reduced serum levels of petroselaidic acid and behenic acid. Transcriptome analyses revealed 38 differentially expressed genes (DEG) in oocytes and 71 DEG in cumulus cells of L (n = 3) compared with M/G group (n = 3). Network analysis connected serum fatty acid changes to downstream effects including reduced inflammatory response and mitochondrial membrane depolarization, increased production of reactive oxygen species, and functions related to fatty acid metabolism and cytoplasmic organization in oocytes. These effects were associated with predicted effects on signaling in oocytes through calcium, insulin, O-GlcNAcase (OGA), fibroblast growth factor receptor 4 (FGF4R), peroxisome proliferator activated receptor gamma coactivator 1 α (PPARGC1A), and phospholipase D2 (PLD2) pathways, with a connection to the cumulus cell via calcium signaling. These results connect BC loss following parturition to changes in serum lipid levels, and changes potentially affecting oocyte quality; thus, these results provide new insight into mechanism of reduced fertility.

Establishment of a vascular system within the corpus luteum (CL) is critical for progesterone (P4) secretion. Measurement of luteal blood flow (LBF) may be a feasible way to determine luteolysis induced with cloprostenol sodium (CLO). Our overall objective was to establish timelines to assess luteolysis via Doppler ultrasonography. Estrous cycles were synchronized in 11- to 12-mo-old Holstein heifers (n = 37). Heifers were injected CLO at a random stage of the estrous cycle 12 d from treatment. Gonadotropin-releasing hormone (GnRH) was administered 2 (d −10) and 8 d (d −4) after the initial CLO. This satisfied the study objective of inducing simultaneous presence of CL at d 4 of development (D4 CL) and CL at d 10 of development (D10 CL) on the day of treatment with different CLO doses (hereafter referred to as d 0). Heifers were randomly assigned to 1 of 5 treatments on d 0: negative control (NC) consisting of no treatment with CLO (n = 8); a quarter dose of CLO (0.125 mg; n = 8); half dose of CLO (0.25 mg; n = 8); full dose of CLO (0.5 mg; n = 8); or positive control (PC) consisting of 4 doses of 0.5 mg of CLO at 24-h intervals starting at d 0 (n = 5). Data collection was performed at d 0 (before and 1 h after treatment) and 2, 4, 6, and 8 d following treatment, to determine luteal volume (LV), LBF, and circulating concentrations of P4. Both NC and PC were efficient in mimicking physiological scenarios that occur during normal luteal development and luteolysis. Heifers that received PC had complete LBF disappearance of both D4 and D10 CL between d 2 and 4 after the first of 4 CLO treatments given 24 h apart (average 4.0 ± 0.0 and 3.2 ± 0.7 d, respectively). Complete LBF disappearance was used as a luteolysis marker. Treatment with different doses of CLO did not impair luteal development of the D4 CL. However, concurrent complete LBF disappearance for D10 CL in heifers treated with half (5/8 heifers) and full doses of CLO (8/8 heifers) resulted in less LBF in the half dose, and less LV and LBF in the full-dose treatment, in D4 CL at d 8 post-treatment, compared with NC. Treatment with various doses of CLO induced an acute increase in LBF 1 h after treatment, regardless of dose in D10 but not in D4 CL. We found a lack of dose response in LV reduction of D4 and D10 CL. Interestingly, LV of the D10 CL decreased in untreated NC between d 0 and 8 after treatment (d 10–18 of luteal development). Assessment with color Doppler ultrasound was sensitive enough to identify dose-response patterns in Holstein heifers (absence, partial, or complete luteolysis) following various doses of CLO. Variability in time to complete LBF disappearance of mature D10 CL following a full dose of CLO limits the use of Doppler ultrasonography to detect luteolysis at a single time point following treatment.