Genome-wide association and genomic prediction for a reproductive index summarizing fertility outcomes in U.S. Holsteins.

Subfertility represents one major challenge to enhancing dairy production and efficiency. Herein, we use a reproductive index (RI) expressing the predicted probability of pregnancy following artificial insemination (AI) with Illumina 778K genotypes to perform single and multi-locus genome-wide association analyses (GWAA) on 2,448 geographically diverse U.S. Holstein cows and produce genomic heritability estimates. Moreover, we use genomic best linear unbiased prediction (GBLUP) to investigate the potential utility of the RI by performing genomic predictions with cross validation. Notably, genomic heritability estimates for the U.S. Holstein RI were moderate (h2 = 0.1654 ± 0.0317-0.2550 ± 0.0348), while single and multi-locus GWAA revealed overlapping quantitative trait loci (QTL) on BTA6 and BTA29, including the known QTL for the daughter pregnancy rate (DPR) and cow conception rate (CCR). Multi-locus GWAA revealed seven additional QTL, including one on BTA7 (60 Mb) which is adjacent to a known heifer conception rate (HCR) QTL (59 Mb). Positional candidate genes for the detected QTL included male and female fertility loci (i.e. spermatogenesis and oogenesis), meiotic and mitotic regulators, and genes associated with immune response, milk yield, enhanced pregnancy rates, and the reproductive longevity pathway. Based on the proportion of the phenotypic variance explained (PVE), all detected QTL (n = 13; P ≤ 5e - 05) were estimated to have moderate (1.0% < PVE ≤ 2.0%) or small effects (PVE ≤ 1.0%) on the predicted probability of pregnancy. Genomic prediction using GBLUP with cross validation (k = 3) produced mean predictive abilities (0.1692-0.2301) and mean genomic prediction accuracies (0.4119-0.4557) that were similar to bovine health and production traits previously investigated.

A 100-Year Review: Historical development of female reproductive physiology in dairy cattle.

The objective of this historical review in female reproductive physiology is to encapsulate major advancements since the founding of the American Dairy Science Association in 1906. The emphasis is on landmark publications in the Journal of Dairy Science since its first volume in 1917. A historical perspective and inferences are made in forecasting evolution of female reproduction and links between physiology of reproduction and the mammary gland. Subsequent sections are focused into main physiological categories and the temporal advancements within these physiological windows. Time points of understanding vary considerably within categories due to various advancements in technology, biological techniques, experimental design, data collection, statistical analyses, and computational forecasting. The physiological windows examined are hypothalamic and pituitary control of the ovary related to estrous behavior and cycle; differential control of the corpus luteum and uterus as influenced by cycling and pregnancy statuses; peripartum and postpartum programing of reproduction; and scientific foundation for the next century. The importance of interdisciplinary programs and integration of reproduction, nutrition/metabolism, genetics, health, and management are emphasized. The modern dairy cow of 2017 exemplifies excellence in both reproductive and lactational performance and is the foundation biological model for the next century.

Chronicling the discovery of interferon tau.

It has been 38 years since a protein, now known as interferon tau (IFNT), was discovered in ovine conceptus-conditioned culture medium. After 1979, purification and testing of native IFNT revealed its unique antiluteolyic activity to prevent the regression of corpora lutea on ovaries of nonpregnant ewes. Antiviral, antiproliferative and immunomodulatory properties of native and recombinant IFNT were demonstrated later. In addition, progesterone and IFNT were found to act cooperatively to silence expression of classical interferon stimulated genes in a cell-specific manner in ovine uterine luminal and superficial glandular epithelia. But, IFNT signaling through a STAT1/STAT2-independent pathway stimulates expression of genes, such as those for transport of glucose and amino acids, which are required for growth and development of the conceptus. Further, undefined mechanisms of action of IFNT are key to a servomechanism that allows ovine placental lactogen and placental growth hormone to affect the development of uterine glands and their expression of genes throughout gestation. IFNT also acts systemically to induce the expression of interferon stimulated genes that influence secretion of progesterone by the corpus luteum. Finally, IFNT has great potential as a therapeutic agent due to its low cytotoxicity, anti-inflammatory properties and effects to mitigate diabetes, obesity-associated syndromes and various autoimmune diseases.

Effects of supplementation with docosahexaenoic acid on reproduction of dairy cows.

The objectives were to determine the effects of supplementing docosahexaenoic acid (DHA)-rich algae on reproduction of dairy cows. Holstein cows were assigned randomly to either a control (n = 373) or the same diet supplemented daily with 100 g/cow of an algae product containing 10% DHA (algae, n = 366) from 27 to 147 days postpartum. Measurements included yields of milk and milk components, fatty acids (FA) profiles in milk fat and plasma phospholipids, resumption of ovulation by 57 days postpartum, pregnancy per artificial insemination (AI) and expression of interferon-stimulated genes in leukocytes. Feeding algae increased resumption of estrous cyclicity (77.6 vs 65.9%) and pregnancy at first AI (47.6 vs 32.8%) in primiparous cows. Algae increased pregnancy per AI in all AI in both primiparous and multiparous cows (41.6 vs 30.7%), which reduced days to pregnancy by 22 days (102 vs 124 days) compared with control cows. Pregnant cows fed algae had greater expression of RTP4 in blood leukocytes compared with those in pregnant control cows. Feeding algae increased the incorporation of DHA, eicosapentaenoic acid, conjugated linoleic acid isomers cis-9 trans-11, trans-10 cis-12 and total n-3 FA in phospholipids in plasma and milk fat. Yields of milk and true protein increased by 1.1 kg/day and 30 g/day respectively, whereas fat yield decreased 40 g/day in algae compared with that in control. Supplementing DHA-rich algae altered the FA composition of lipid fractions and improved reproduction in dairy cows. The benefits on reproduction might be mediated by enhanced embryo development based on changes in interferon-stimulated gene expression.

Role of lipids on elongation of the preimplantation conceptus in ruminants.

Elongation of the preimplantation conceptus is a prerequisite for successful pregnancy in ruminants and depends on histotroph secretion by the endometrium. Lipids are an essential component of the histotroph, and recent studies indicate that lipids have important roles in the elongation phase of conceptus development. The onset of elongation is marked by dynamic changes in the transcriptome of trophectoderm cells, which are associated with lipid metabolism. During elongation, the trophectoderm increases transcript expression of genes related to uptake, metabolism and de novo biosynthesis of fatty acids and prostaglandins. Expression of the gene PPARG increases substantially, and activation of the transcription factor PPARG by binding of lipid ligands appears to be crucial for the coordination of cell biology during elongation. Lipids accumulated in the epithelial cells of the endometrium during diestrus are likely the most important source of fatty acids for utilization by the conceptus and become available in the uterine lumen through exporting of exosomes, microvesicles, carrier proteins and lipoproteins. Targeting of uterine lipid metabolism and PPARG activity during preimplantation conceptus development through nutraceutical diets may be a good strategy to improve pregnancy survival and reproductive efficiency in ruminants.

Biology of Preimplantation Conceptus at the Onset of Elongation in Dairy Cows.

The objectives of this study were to characterize changes in transcriptome of preimplantation conceptuses at the onset of elongation and associated changes in uterine histotroph composition and endometrial physiology. Lactating dairy cows (n = 160) had their ovulation synchronized by artificial insemination (study Day 0). On Day 15, uteri were flushed and endometrium tissue collected. Recovered conceptuses were classified based on morphology/length as ovoid (1-4 mm), tubular (5-19 mm), and filamentous (20-60 mm). Total RNA (n = 48) was subjected to transcriptome analysis. The uterine fluid (n = 30) was evaluated by mass spectrophotometry. Transcriptome analyses revealed drastic changes in the transition from ovoid to tubular and from tubular to filamentous. Differentially expressed genes were associated with cellular movement, cell-to-cell signaling, cellular assembly and organization, lipid metabolism, small molecule biochemistry, and molecular transport. Specific changes included reorganization of cytoskeleton and cell migration, arginine metabolism, growth factors signaling, and lipid metabolism. Functional analysis revealed fatty acids and peroxisome proliferator activated receptor gamma as upstream regulators of transcriptome changes. Expression of PPARG increased 17-fold during the onset of elongation and was highly correlated with genes involved in lipid metabolism. The histotroph was rich in amino acids, lipids, saccharides, and other intermediate metabolites, and important changes in composition occurred in the presence of a conceptus. Pregnancy had a major impact on the concentrations of important lipids in the uterine fluid and expression of genes in the endometrium. Collectively, conceptus elongation involves remarkable changes in transcriptome, composition of the histotroph, and endometrial physiology, which help elucidate important events in uterine and conceptus biology at the onset of elongation.

Invited review: Recommendations for reporting intervention studies on reproductive performance in dairy cattle: Improving design, analysis, and interpretation of research on reproduction.

Abundant evidence from the medical, veterinary, and animal science literature demonstrates that there is substantial room for improvement of the clarity, completeness, and accuracy of reporting of intervention studies. More rigorous reporting guidelines are needed to improve the quality of data available for use in comparisons of outcomes (or meta-analyses) of multiple studies. Because of the diversity of factors that affect reproduction and the complexity of interactions between these, a systematic approach is required to design, conduct, and analyze basic and applied studies of dairy cattle reproduction. Greater consistency, clarity, completeness, and correctness of design and reporting will improve the value of each report and allow for greater depth of evaluation in meta-analyses. Each of these benefits will improve understanding and application of current knowledge and better identify questions that require additional modeling or primary research. The proposed guidelines and checklist will aid in the design, conduct, analysis, and reporting of intervention studies. We propose an adaptation of the REFLECT (Reporting Guidelines for Randomized Controlled Trials for Livestock and Food Safety) statement to provide guidelines and a checklist specific to reporting intervention studies in dairy cattle reproduction. Furthermore, we provide recommendations that will assist investigators to produce studies with greater internal and external validity that can more often be included in systematic reviews and global meta-analyses. Such studies will also assist the development of models to describe the physiology of reproduction.

Meta-analysis of the effects of sometribove zinc suspension on the production and health of lactating dairy cows.

OBJECTIVE: To provide an updated evaluation of the efficacy and safety of sometribove zinc suspension (rbST-Zn), a form of recombinant bovine somatotropin, in lactating dairy cows. DESIGN: Meta-analysis. SAMPLE: 26 studies published in peer-reviewed journals or reviewed by a regulatory agency. PROCEDURES: To be included, a study had to involve the use of the rbST-Zn formulation available to US producers in accordance with the label instructions for treatment initiation (57 to 70 days postpartum), dose (500 mg, q 14 d), and route (SC). RESULTS: For cows treated with rbST-Zn, mean milk, 3.5% fat-corrected milk, fat, and protein yields were increased by 4.00, 4.04, 0.144, and 0.137 kg/d (8.8, 8.89, 0.32, and 0.30 lb/d), respectively; however, the concentration of milk components did not change. Pregnancy proportion for the first 2 breeding cycles was increased by 5.4%, and pregnancy proportion for the duration of the trial was reduced by 5.5% for rbST-Zn-treated cows, compared with proportions for untreated cows. Mean body condition score (1 to 5 scale) was reduced by 0.06 points during the period of rbST-Zn use for treated cows. Administration of rbST-Zn had no effect on milk somatic cell count, the number of days to pregnancy, or inseminations per pregnancy; rates of fetal loss, twins, cystic ovaries, clinical lameness, lameness lesions, or traumatic lesions of the integumentary system; and odds of clinical mastitis or culling. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that rbST-Zn administration to dairy cows effectively increases milk production with no adverse effects on cow health and well-being.

Low doses of bovine somatotropin enhance conceptus development and fertility in lactating dairy cows.

Objectives were to evaluate the effects of administering either one or two low doses of slow-release recombinant bovine somatotropin (bST) on hormone concentrations, conceptus development, and fertility in dairy cows. Cows from two farms were detected in estrus on or after 50 days postpartum (n = 1483), inseminated, and enrolled in the study (Day 0). Within farm, cows were blocked by parity and assigned randomly to receive a single placebo injection at insemination (control), a single injection with 325 mg of bST at insemination (S-bST), or two injections with 325 mg of bST administered on Days 0 and 14 (T-bST). From a subset of cows, blood was collected twice weekly from Day 0 to 42 for determination of hormone concentrations and on Day 19 for isolation of leucocytes and analysis of transcript abundance of selected interferon-stimulated genes. Pregnancy was diagnosed on Days 31 and 66, and ultrasonographic morphometry of the conceptus was performed on Days 34 and 48 in a subset of cows. Cows that received T-bST had increased plasma concentrations of GH and IGF1 for 4 wk, increased mRNA expression of ISG15 and RTP4 in leukocytes, earlier rise in the pregnancy-specific protein B in plasma of pregnant cows, increased conceptus size, and enhanced fertility. Cows that received S-bST had increased concentrations of GH and IGF1 for only 2 wk and it was insufficient to alter conceptus development and fertility. In conclusion, supplementation with low doses of bST during the pre- and peri-implantation periods enhanced conceptus development, reduced embryonic losses, and improved fertility in dairy cows.

Roles of conceptus secretory proteins in establishment and maintenance of pregnancy in ruminants.

Reproduction in ruminant species is a highly complex biological process requiring a dialogue between the developing conceptus (embryo-fetus and associated placental membranes) and maternal uterus which must be established during the peri-implantation period for pregnancy recognition signaling and regulation of gene expression by uterine epithelial and stromal cells. The uterus provide a microenvironment in which molecules secreted by uterine epithelia and transported into the uterine lumen represent histotroph, also known as the secretome, that are required for growth and development of the conceptus and receptivity of the uterus to implantation by the elongating conceptus. Pregnancy recognition signaling as related to sustaining the functional lifespan of the corpora lutea, is required to sustain the functional life-span of corpora lutea for production of progesterone which is essential for uterine functions supportive of implantation and placentation required for successful outcomes of pregnancy. It is within the peri-implantation period that most embryonic deaths occur in ruminants due to deficiencies attributed to uterine functions or failure of the conceptus to develop appropriately, signal pregnancy recognition and/or undergo implantation and placentation. The endocrine status of the pregnant ruminant and her nutritional status are critical for successful establishment and maintenance of pregnancy. The challenge is to understand the complexity of key mechanisms that are characteristic of successful reproduction in humans and animals and to use that knowledge to enhance fertility and reproductive health of ruminant species in livestock enterprises.

Response to GnRH on day 6 of the estrous cycle is diminished as the percentage of Bos indicus breeding increases in Angus, Brangus, and Brahman x Angus heifers.

Angus (n=6), Brangus (5/8 Angus x 3/8 Brahman, n=6), and Brahman x Angus (3/8 Angus x 5/8 Brahman, n=6) heifers exhibiting estrous cycles at regular intervals were used to determine if the percentage of Bos indicus breeding influenced the secretory patterns of LH in response to a GnRH treatment on Day 6 of the estrous cycle. Heifers were pre-synchronized with a two-injection PGF(2 alpha) protocol (25 mg i.m. Day -14 and 12.5 mg i.m. Day -3 and -2 of experiment). Heifers received 100 microg GnRH i.m. on Day 6 of the subsequent estrous cycle. Blood samples were collected at -60, -30, and -1 min before GnRH and 15, 30, 60, 90, 120, 150, 180, 240, 300, 360, 420, and 480 min after GnRH to determine concentrations of serum LH. Estradiol concentrations were determined at -60, -30, and -1 min before GnRH. On Day 6 and 8, ovaries were examined by ultrasonography to determine if ovulation occurred. On Day 13, heifers received 25 mg PGF(2 alpha) i.m. and blood samples were collected daily until either the expression of estrus or Day 20 for heifers not exhibiting estrus to determine progesterone concentrations. There was no effect (P>0.10) of breed on ovulation rate to GnRH as well as size of the largest follicle, mean estradiol, and mean corpus luteum volume at GnRH. Mean LH was greater (P<0.05) for Angus (7.0+/-0.8 ng/mL) compared to Brangus (4.6+/-0.8 ng/mL) and Brahman x Angus (2.9+/-0.8 ng/mL), which were similar (P>0.10). Mean LH peak-height was similar (P>0.10) for Brangus (13.9+/-3.4 ng/mL) compared to Angus (21.9+/-3.4 ng/mL) and Brahman x Angus (8.0+/-3.4 ng/mL), but was greater (P<0.05) for Angus compared to Brahman x Angus. Interval from GnRH to LH peak was similar (P>0.10) between breeds. As the percentage of Bos indicus breeding increased the amount of LH released in response to GnRH on Day 6 of the estrous cycle decreased.

Evaluation of the efficacy of prophylactic hoof health examination and trimming during midlactation in reducing the incidence of lameness during late lactation in dairy cows.

OBJECTIVE: To assess the efficacy of prophylactic hoof health examination and trimming during midlactation at reducing the incidence of lameness during late lactation in dairy cows. DESIGN: Randomized field trial. ANIMALS: 333 Holstein cows. PROCEDURES: Cows without apparent lameness were randomly allocated into 1 of 2 groups approximately 204 days after calving. Cows allocated to the treatment group (n = 161) were examined on a tilt table for diagnosis and underwent hoof-trimming procedures, if needed, for treatment of hoof disorders or lesions. Cows in the control group (n = 172) were not examined. Cows were assigned a locomotion score weekly for 28 weeks after allocation to a group. The number of cows classified as lame during late lactation (approx 205 to 400 days after calving) was compared between groups to assess the efficacy of prophylactic examination and trimming. RESULTS: Incidence of lameness during late lactation was 24% in cows in the control group and 18% in cows in the treatment group. CONCLUSIONS AND CLINICAL RELEVANCE: The 25% decrease in number of new cases of lameness in cows undergoing prophylactic hoof health examination and trimming during midlactation may be relevant for the well-being of dairy cows and should not represent a major economic burden to producers.

Peroxisome proliferator-activated receptor (PPAR) expression in cultured bovine endometrial cells and response to omega-3 fatty acid, growth hormone and agonist stimulation in relation to series 2 prostaglandin production.

The peroxisome proliferator-activated receptors (PPARs) are a family of nuclear transcription factors thought to act as receptors for polyunsaturated fatty acids and to reduce production of series 2 prostaglandins (PG). The objectives of the current study were to characterize PPAR expression and the prostaglandin synthetic activity of cultured bovine endometrial cells in response to known PPAR ligands, as well as to key stimulators and inhibitors of series 2 prostaglandin secretion. PPARalpha and PPARdelta, but not PPARgamma, mRNAs are expressed in the BEND cell line regardless of treatment. Under resting conditions, PPARalpha mRNA levels increase in response to growth hormone (P < 0.05). In cells stimulated with PdBu, growth hormone depresses PPARalpha mRNA levels, regardless of whether cells also are treated with IFNtau. In contrast, PPARdelta mRNA levels are increased by exposure to PdBu, eicosapentanoic acid and IFNtau, and these effects are additive. PPAR mRNA levels are not predictive of prostaglandin accumulation. Agonist activation of PPARalpha, PPARdelta or PPARgamma augments PdBu-induced increases in prostaglandin H synthase-2 mRNA and media accumulation of prostaglandins F2alpha and E2. Treatment with the PPARalpha/delta agonist carbaprostacyclin, but not the PPARalpha agonist Wy14643 or PPARgamma agonist ciglitazone, completely reverses the IFNtau suppression of prostaglandin synthesis. In conclusion, PPARalpha and PPARdelta function in the response of bovine endometrium to growth hormone and long chain omega-3 polyunsaturated fatty acids.

Comparison of the calving-to-conception interval in dairy cows with different degrees of lameness during the prebreeding postpartum period.

OBJECTIVE: To compare calving-to-conception intervals among cows classified as nonlame, moderately lame, or lame during the prebreeding postpartum period and to examine the relationship between severity of lameness and time to conception in cows that were classified as lame. DESIGN: Longitudinal study. ANIMALS: 499 Holstein cows. PROCEDURE: Cows in the prebreeding postpartum period were classified as nonlame, moderately lame, or lame by use of a 6-point locomotion scoring system. Time to conception (days) was compared among cows. A low, medium, or high cumulative locomotion score was assigned to lame cows, and time to conception among those cows was compared. Cows classified as lame were examined on a tilt table for diagnosis and treatment of lameness. RESULTS: 154 (31%), 214 (43%), and 131 (26%) cows were classified as nonlame, moderately lame, and lame, respectively. Most cows classified as lame had laminitis (54%) or disorders of the claw (33%). Median time to conception was 36 to 50 days longer in lame cows than in nonlame cows. Among lame cows, the median time to conception was 66 days longer in cows with high cumulative locomotion scores than in cows with low scores. CONCLUSION AND CLINICAL RELEVANCE: Nonlame cows became pregnant more quickly than lame cows. Lame cows with low cumulative locomotion scores during the prebreeding postpartum period became pregnant sooner than lame cows with high scores. Early diagnosis and intervention may mitigate the effects of lameness and improve reproductive performance in lame dairy cows.

Comparison of milk yield in dairy cows with different degrees of lameness.

OBJECTIVE: To compare milk yield among cows classified as nonlame, moderately lame, and lame and to examine the relationship between severity of lameness and milk yield in cows classified as lame during the first 100 days after parturition. DESIGN: Longitudinal study. ANIMALS: 465 Holstein cows. PROCEDURE: Cows were examined weekly during the first 100 days after parturition and assigned a lameness score by use of a 6-point locomotion scoring system (ie, 0 to 5). Milk yield was compared among cows classified as nonlame, moderately lame, and lame. Among cows classified as lame (locomotion score > or = 4), milk yield was compared for cows with low, medium, and high cumulative locomotion scores. Cows classified as lame were further examined on a tilt table for diagnosis and treatment of lameness. RESULTS: 84 (18%), 212 (46%), and 169 (36%) cows were classified as nonlame, moderately lame, and lame, respectively. Among cows in their second or later lactations, milk yield in lame cows was significantly lower than that in moderately lame and nonlame cows. In addition, among cows classified as lame, milk yield was significantly lower in cows with high locomotion scores during the first 100 days after parturition, compared with cows with low scores. Most (58%) cows classified as lame had laminitis. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate a linear relationship between increasing degree of lameness and decreasing milk yield among cows in their second or later lactations. The locomotion scoring system used in this study may be a useful management tool that veterinarians and dairy farmers could adopt for early detection of lameness in dairy cows.

Efficacy of either a single or split treatment of PGF2alpha after a 14 day melengestrol acetate treatment to synchronize estrus and induce luteolysis in Bos indicus x Bos taurus heifers.

Two experiments evaluated a modified delivery of prostaglandin F2alpha (PGF2alpha) after a melengestrol acetate (MGA) treatment in Angus and Bos indicus x Bos taurus (BI) heifers. Experiment 1 was replicated three times with yearling BI heifers (n = 695). Heifers received MGA (0.5 mg head(-1) day(-1)) for 14 days. In Replications 1 and 2, heifers received either 25 mg of PGF2alpha im 19 days after MGA (single) or 12.5 mg of PGF2alpha im 19 and 20 days after MGA (split). In Replication 3, heifers received the same treatments, with PGF2alpha initiated either 18 or 19 days after MGA. Estrus was detected for 72 h after PGF2alpha, with AI commencing 8-12 h after a detected estrus. Heifers not observed in estrus by 72 h were timed-AI concomitant with GnRH (100 microg im). Heifers from Replication 2 (n = 146) had blood samples collected at the initial PGF2alpha and at timed-AI to determine corpus luteum (CL) regression by evaluating plasma progesterone concentrations. The interval from MGA withdrawal to PGF2alpha did not have a significant effect on any variable in Replication 3 and there were no treatment by replication effects for any variables, therefore data were pooled. Modifying the PGF2alpha treatment from a single treatment to two treatments on consecutive days increased (P < 0.05) 72 h estrous response (43.2% versus 50.1%), timed-AI (23.9% versus 33.5%) and total-AI pregnancy rates (34.5% versus 42.5%), and CL regression (79.1% versus 92.5%), respectively. In Experiment 2, yearling Angus (n = 66) and 2-year-old BI (n = 68) heifers were synchronized as per Experiment 1 (with the initial PGF2alpha 19 days after MGA). Neither breed nor PGF2alpha treatment effected (P > 0.05) 72 h estrous response, total-AI pregnancy rate, or CL regression rate. In conclusion, treating yearling BI heifers with split treatments of PGF2alpha (given on two consecutive days) improved estrous response and pregnancy rates by increasing PGF2alpha-induced luteolysis.

Ovarian and endocrine responses associated with the treatment of cystic ovarian follicles in dairy cows with gonadotropin releasing hormone and prostaglandin F2alpha, with or without exogenous progesterone.

The objectives of this observational study were to document ovarian and endocrine responses associated with the treatment of cystic ovarian follicles (COFs) in dairy cows, using gonadotropin releasing hormone (GnRH) and prostaglandin F2alpha (PGF) with or without exogenous progesterone. A secondary objective was to determine pregnancy establishment following synchronization of ovulation and timed insemination in cows diagnosed with COFs. In trial I, 18 Holstein cows diagnosed with COFs received 2 injections of 100 microg GnRH, 9 d apart, with 25 mg PGF given 7 d after the 1st GnRH. A new follicle developed in all 18 cows after the 1st GnRH, and 83% of cows ovulated following the 2nd GnRH. Cows were inseminated 16 h after the 2nd GnRH. Of the 17 cows available for pregnancy diagnosis, 7 were confirmed pregnant. In trial II, 8 cows with COFs received GnRH and an intravaginal progesterone device (CIDR) concurrently, then PGF 7 d later. The CIDR was removed 2 d after PGF administration. Plasma estradiol concentrations declined following CIDR insertion. In all cows, a new follicle developed following GnRH treatment; estradiol-surge and estrus occurred spontaneously after CIDR-removal. Seven of 8 cows ovulated the new follicle. In dairy cows diagnosed with COFs, treatment with GnRH followed by PGF 7 d later, with or without exogenous progesterone, resulted in the recruitment of a healthy new follicle; synchronization of ovulation and timed insemination resulted in a 41% pregnancy rate.

Interferon-tau induces degradation of prostaglandin H synthase-2 messenger RNA in bovine endometrial cells through a transcription-dependent mechanism.

A series of experiments were undertaken to examine the effects of interferon (IFN)-tau on regulation of prostaglandin H synthase (PGHS)-2 mRNA in bovine endometrial (BEND) cells as a means to elucidate the actions of IFN-tau to maintain pregnancy. The objective was to determine if IFN-tau mediates posttranscriptional regulation of PGHS-2 mRNA. Cells were treated with phorbol 12,13-dibutyrate (PdBu) for 3 h to induce PGHS-2 mRNA expression. Actinomycin D (0 or 1 microg/ml) or the p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580 (1 microM), were added at 3 h, followed by addition of IFN-tau (0 or 50 ng/ml) at 3.5 h and extraction of RNA at 4.5 h. The concentrations of PGHS-2 mRNA were stable between 3 and 4.5 h regardless of actinomycin D. Simultaneous treatment of PdBu-treated cells with actinomycin D and SB203580 (1 microM) decreased PGHS-2 mRNA. Addition of IFN-tau (50 ng/ml) reduced PGHS-2 mRNA, which was not observed when actinomycin D was present. Concurrent treatments of cells with SB203580 and IFN-tau (5 ng/ml) decreased concentrations of PGHS-2 mRNA in an additive manner. Although IFN-tau reduced PGHS-2 mRNA concentrations, phosphorylation of p38 MAPK was induced by IFN-tau, PdBu, and PdBu combined with IFN-tau after 10 min of treatment. Both the p38 MAPK inhibitor and IFN-tau decreased prostaglandin F(2alpha) secretion, and decreases were additive when the two were given together. In summary, activation of p38 MAPK by PdBu is required for continued presence of PGHS-2 mRNA and secretion of prostaglandin F(2alpha) in BEND cells. Interferon-tau mediates a transcription-dependent mechanism, which induces degradation of PGHS-2 mRNA. However, the consequences of an IFN-tau-induced activation of p38 MAPK warrant further investigation, because inhibition of p38 MAPK caused a degradation of PGHS-2 mRNA.

Effect of resynchronization with GnRH on day 21 after artificial insemination on pregnancy rate and pregnancy loss in lactating dairy cows.

The objectives of the present study were to determine the effects of resynchronization with GnRH on Day 21 after artificial insemination (AI) on pregnancy rate and losses of pregnancy in lactating dairy cows. Holstein cows (n=585) on two dairy farms were assigned to one of two treatments in a randomized complete block design. On Day 21 after a pre-enrollment AI, animals assigned to the resynchronization (RES) group received 100 microg of GnRH i.m., whereas animals in the control (CON) group received no treatment. All animals were examined ultrasonographically on Days 21 and 28 after AI, and blood samples were taken for progesterone measurement on Day 21. Pregnancy was diagnosed on Day 28 and reconfirmed 14 days later. Nonpregnant cows on Day 28 were inseminated using timed AI after the completion of the Ovsynch protocol 10 and 17 days after enrollment in the study for RES and CON groups, respectively. Progesterone concentration > or =2.35 ng/ml was used as an indicator of pregnancy on Day 21. For RES and CON cows, pregnancy rate at Days 21 (70.9% versus 73.0%, P<0.56), 28 (33.1% versus 33.6%; P<0.80) and 42 (27.0% versus 26.8%; P<0.98) after the pre-enrollment AI did not differ. Administration of GnRH on Day 21 after AI had no effect on pregnancy loss in RES and CON groups from days 21 to 28 (53.2% versus 53.5%; P<0.94) and days 28 to 42 (17.9%; P<0.74) after AI. Pregnancy rate after the resynchronization period was similar for both treatment groups. Resynchronization with GnRH given on Day 21 after AI for initiation of a timed AI protocol prior to pregnancy diagnosis does not affect pregnancy rate and pregnancy loss in lactating dairy cows.