Reproductive tract disease in Irish grazing dairy cows: Retrospective observational study examining its association with reproductive performance and accuracy of 2 diagnostic tests.

The detection of reproductive tract disease (RTD) 3 wk postpartum is important because of its effect on subsequent reproductive outcomes. Numerous methods for the diagnosis of RTD are described, some of which are more practical and instantaneous in terms of diagnosis. Two of these methods involve identification of purulent vaginal discharge (PVD) and evidence of ultrasonographic uterine changes indicative of endometritis (UE). The objectives of our retrospective observational study were (1) to assess the association of PVD or UE score at the prebreeding examination (PBE) with the hazard of pregnancy within the subsequent breeding season; (2) to determine the test sensitivity (Se) and specificity (Sp) at the point of sampling of both tests using a Bayesian latent class model; and (3) to determine the effect of varying positivity thresholds on test accuracy. To achieve these objectives, we analyzed an initial data set of 5,049 PBE from 2,460 spring-calved cows in 8 herds between 2014 and 2018. Each PBE was conducted once between 25 and 86 d in milk. At each PBE, vaginal discharge was obtained with a Metricheck device (Simcro) whereas uterine contents were assessed using transrectal ultrasonography. Purulent vaginal discharge was scored on a scale of 0 to 3 depending on discharge character, and UE was scored on a scale of 0 to 4 depending on the presence and consistency of intraluminal fluid. Cows with scores of ≥2 in either test had received treatment. Fertility data were available from 4,756 PBE after data exclusion. The association between PVD or UE score at the PBE and subsequent hazard of pregnancy was analyzed using a Cox proportional hazards model. Cows with a PVD score of 2 or 3 were less likely to conceive than cows with a PVD score 0 [score 2 hazard ratio (HR) = 0.74; 95% confidence interval (CI): 0.59-0.94; score 3 HR = 0.65; 95% CI: 0.51-0.84]. Cows with a UE score of 1, 2, 3, or 4 were less likely to conceive than cows with a UE score of 0 (score 1 HR = 0.82; 95% CI: 0.73-0.93; score 2 HR = 0.79; 95% CI: 0.62-1.00; score 3 HR = 0.43; 95% CI: 0.43-0.90; score 4 HR = 0.39; 95% CI: 0.26-0.58). To determine the Se and Sp of PVD or UE score for diagnosis of RTD at the time of PBE, a Bayesian latent class model was fitted on 2,460 individual cow PBE. Flat priors were used for the Se and Sp of UE, whereas informative priors were used for PVD Se (mode = 65%, 5th percentile = 45%) and Sp (mode = 90%, 5th percentile = 80%) and RTD prevalence (mode = 20%, 5th percentile = 10%). Posterior estimates (median and 95% Bayesian probability intervals; BPI) were obtained using 'rjags' (R Studio). The optimal test thresholds (PVD and UE score ≥1) were selected by assessing the effect of different thresholds on test estimates and using a misclassification cost analysis. Based on these, median (95% BPI) Se for PVD and UE score ≥1 were 44% (29-60%) and 67% (33-100%), respectively. Median Sp for PVD and UE score ≥1 were 90% (86-93%) and 91% (86-93%), respectively. Higher scores in both tests were associated with impaired fertility, and UE scoring with a threshold of ≥1 had the highest test Se and Sp estimates although test Se was conditional on days in milk when the PBE occurred.

Cow-level risk factors for reproductive tract disease diagnosed by 2 methods in pasture-grazed dairy cattle in Ireland.

The cow-side diagnosis of reproductive tract disease (RTD) involves identifying the presence of purulent vaginal discharge (PVD) and ultrasonographic endometritis (UE). The objectives of our study were to obtain prevalence estimates for RTD diagnosed by 2 methods (PVD and UE scoring) and to investigate the risk factors for increased probability of RTD if these methods are used in isolation or in combination. Our retrospective observational cohort study tested the hypothesis that RTD assessed by 2 methods would have similar risk factors, and that those would be mainly cow- and calving-related factors. We analyzed data from 5,049 pre-breeding examinations (PBE) from 2,460 spring-calved cows on 8 farms between 2014 and 2018. Cow-related details assessed were days in milk at PBE, breed, lactation number, dry period length, body condition score at calving and PBE, 305-d milk yield, predicted transmitting ability for production and fertility, the presence of a corpus luteum at PBE, and positive diagnosis the previous year. Calving details assessed were type of sire, calf sex, twinning, stillbirth, calving difficulty score, and retained fetal membranes. We conducted statistical analyses using 4 multivariable logistic regression models to identify the risk of RTD diagnosed by (1) PVD in isolation, (2) UE in isolation, (3) the presence of either PVD or UE; and (4) the presence of both PVD and UE. We accounted for herd, cow, and year as random effects in all 4 models. The overall prevalence of RTD in models 1, 2, 3, and 4 were 7.5, 6.7, 11.6, and 2.6%, respectively. Days in milk at PBE, the interaction between days in milk and retained fetal membranes, twinning, and the predicted transmitting ability for calving interval were consistently significant risk factors for positive scores in all 4 models. Considerable calving difficulty [adjusted odds ratio (AOR) = 13.64], Holstein Friesian dam breed (AOR = 2.58), first lactation (AOR = 2.39), and body condition score at PBE (AOR = 1.64) were risk factors for a positive PVD score but not for a positive UE score. Fifth lactation (AOR = 1.69), a beef-sired calf (AOR = 1.46), and the absence of a corpus luteum at PBE (AOR = 1.57) were risk factors for a positive UE score but not for a positive PVD score. These results support the hypothesis that most of the risk factors for PVD and UE are the same but some are distinctly different, implying that in some instances the 2 methods diagnose separate components of the RTD complex.

Altered endometrial immune gene expression in beef heifers with retarded embryos.

The aim of the present study was to compare endometrial gene expression profiles in a group of beef heifers yielding viable or retarded embryos on Day 7 after oestrus as a means of potentially explaining differences in embryo survival rates. Heifers were classified as either: (1) viable, when the embryo collected on Day 7 after oestrus was at the correct developmental stage (i.e. morula/early blastocyst); or (2) retarded, when the embryo was arrested at the 2-16-cell stage. The focus of the present study was on genes that were associated with either the pro- or anti-inflammatory immune response. Endometrial gene expression was determined using quantitative real-time polymerase chain reaction analysis. Expression of the ß-defensin (DEFB1), interferon (IFN)-α (IFNA), IFN-γ (IFNG), interleukin (IL)-6 (IL6), IL-10 (IL10), forkhead box P3 (FOXP3) and natural cytotoxicity triggering receptor 1 (NCR1) genes was lower in endometria from viable than retarded heifers. Expression of the nuclear factor of kappa light polypeptide gene enhancer in B cells 1 (NKFB1), transforming growth factor (TGF)-ß (TGFB), IFN-γ-inducible protein 16 (IFI16) and IL-21 (IL21) genes was higher in viable than retarded heifers. We propose that small disturbances in the expression of immune genes in the endometrium on Day 7 after oestrus can have detrimental effects on embryo survival.

The effect of natural and induced calving of beef heifers on stress-related gene expression and maternal health and immunity.

The peri-partum processes can exert stress on a cow on many levels. There is little evidence about acute stress around the calving event and subsequent potential effects for the cows' immunological status or subsequent reproductive health. To investigate this, 55 crossbred recipient beef heifers carrying purebred Simmental embryos were assigned to one of three groups on day 285 of gestation: (i) control (no parturition induction treatment; n = 19); (ii) induction of parturition with corticosteroid (n = 20) and (iii) induction of parturition with corticosteroid plus prostaglandin (n = 16). Interval from induction of parturition to calving and calving ease was recorded. Reproductive tract examinations were conducted on Day 21 (D21) and Day 42, and a sample was obtained for the determination of uterine cytology on D21. Blood samples were taken from the dams two weeks before parturition, one day after parturition (D1) and two weeks after parturition (D14) for gene expression and cortisol and calcium concentration determination. Calves were weighed at birth and subsequently every week until they were 10 weeks of age. A colostrum sample was taken immediately after calving and stored for subsequent Immunoglobin G (IgG) concentration analysis. Data were analysed using ANOVA with posthoc Tukey, Spearman correlation and stepwise backwards linear regression using SAS. Quantitative reverse transcription PCR was performed on the following immune genes: Interleukins IL1a and b, IL2, IL4, IL8, Tumour Necrosis Factor Alpha, Interferon-gamma, Lymphotoxin, Toll-Like Receptor, Nuclear factor of kappa light polypeptide gene enhancer in B cells 1 and 2, glucocorticoid receptor alpha, as well as the neutrophil genes that regulate inflammation: Fas, L-selectin, MMP-9 and BPI. The results show that compared with non-induced contemporaries, induction has no negative effect on dystocia or subsequent calf weight gain but can have a positive effect on colostral IgG concentration. Blood calcium concentrations on both D1 and D14 postcalving are associated with subsequent uterine health. Parturition events were reflected in temporal changes in the expression of the cytokines IFNγ, TNFα, IL1b, IL4, IL8 and Haptoglobin in the dams' blood, all of which are associated with the immune competence of the cow during this period. The conclusion is that induction of calving can have a positive effect on colostral IgG concentration. Calcium concentrations postcalving are associated with subsequent reproductive tract health. Events associated with the peri- and postpartum period are all reflected in temporal changes in immune function-related cytokines.

Changes in the endometrial transcriptome during the bovine estrous cycle: effect of low circulating progesterone and consequences for conceptus elongation.

In cattle, elevated concentrations of circulating progesterone (P4) in the immediate postconception period are associated with advanced conceptus development, while low P4 is implicated as a causative factor in low pregnancy rates observed in dairy cows. This study aimed to: 1) describe the transcriptional changes that occur in the bovine endometrium during the estrous cycle, 2) determine how elevated P4 affects these changes, 3) identify if low P4 alters the expression of these genes, and 4) assess the impact that low P4 has on conceptus development. Relatively few differences occurred in endometrial gene expression during the early luteal phase of the estrous cycle (Day 5 vs. 7), but comparison of endometria from more distant stages of the luteal phase (Day 7 vs. 13) revealed large transcriptional changes, which were significantly altered by exogenous supplementation of P4. Induction of low circulating P4 altered the normal temporal changes in gene expression, and these changes were coordinate with a delay in the down-regulation of the PGR from the LE and GE. Altered endometrial gene expression induced by low P4 was associated with a reduced capacity of the uterus to support conceptus development after embryo transfer on Day 7. In conclusion, the present study provides clear evidence that the temporal changes in the transcriptome of the endometrium of cyclic heifers are sensitive to circulating P4 concentrations in the first few days after estrus. Under low P4 conditions, a suboptimal uterine environment with reduced ability to support conceptus elongation is observed.

Estimation of the true prevalence of inaccurate artificial inseminations in Irish milk recording dairy cows using a Bayesian latent class analysis.

Inaccurate artificial insemination (IAI) refers to an artificial insemination (AI) that is performed when a cow is not in oestrus. IAIs have economic impacts on the dairy industry through of semen wastage or iatrogenic pregnancy loss. However, few studies have quantified the prevalence of IAIs in a population. The primary objective of this prospective study was to estimate the cow-level true prevalence of IAIs in Irish milk recording dairy herds using a latent class model with a Bayesian framework. Milk samples were collected at a milk recording laboratory from 576 dairy cows in 125 herds who had received an AI on the same day they were sampled for routine milk constituent analysis. Milk progesterone (MP4) analysis was conducted on these samples using radioimmunoassay to determine the progesterone concentration. Fertility data (i.e., subsequent calving date) was retrospectively obtained from the Irish National Cattle Breeding Federation for milk sampled cows and an apparent conception (AC) to the sample AI was determined based on an estimated gestational range of 270-290 days. Both tests (MP4 and AC) were used in a latent class model to estimate the true prevalence of IAI. For the MP4 test, a concentration of ≥ 5 ng/mL in whole milk was deemed to be test positive while for the AC test, a cow that did not conceive to the sampled AI was deemed test positive. Prior information for prevalence of IAI was obtained from a literature review while MP4 sensitivity (Se) and specificity (Sp) were obtained from expert opinion. Non-informative priors were used for the Se and Sp of the AC test. Posterior inferences (median and 95 % Bayesian probability intervals; BPI) were obtained using the 'rjags' package in the R statistical software. In the final model, median cow-level true prevalence of IAI was 4.4 % (BPI; 1.7-9.0 %). Median Se and Sp estimates for MP4, were 83.0 % (BPI; 65.0-96.2 %) and were 97.4 % (BPI; 94.6-99.6 %), respectively. Median Se and Sp estimates for AC, were 64.8 % (BPI; 44.5-88.6 %) and 49.8 % (BPI; 45.3-54.1 %), respectively. The present study estimates that the overall cow-level true prevalence of IAI in Irish dairy cows is relatively low. This is the first study to report the cow-level true prevalence of IAI using a Bayesian latent class model.

Characterisation of endometrial gene expression and metabolic parameters in beef heifers yielding viable or non-viable embryos on Day 7 after insemination.

The aim of the present study was to compare the hormonal and metabolic characteristics and endometrial gene expression profiles in beef heifers yielding either a viable or degenerate embryo on Day 7 after insemination as a means to explain differences in embryo survival. Oestrus was synchronised in cross-bred beef heifers (n = 145) using a controlled internal drug release (CIDR)-prostaglandin protocol. Heifers (n = 102) detected in standing oestrus (within 24-48 h after CIDR removal) were inseminated 12-18 h after detection of oestrus (Day 0) with frozen-thawed semen from a single ejaculate of a bull with proven fertility. Blood samples were collected from Day 4 to Day 7 after oestrus to measure progesterone (on Days 4, 5 and 7), insulin and insulin-like growth factor (IGF)-I (on Days 4 and 6) and urea (on Day 7) concentrations. All animals were killed on Day 7. Uterine pH was determined at the time of death. Animals from which an embryo was recovered were classified as either having a viable embryo (morula/blastocyst stage; n = 32) or a retarded embryo (arrested at the two- to 16-cell stage; n = 19). In addition, 14 single-celled unfertilised oocytes were recovered, giving an overall recovery rate of 64%. There was no significant difference in the blood parameters determined or uterine pH at the time of death between heifers with either a viable or retarded embryo. The relative abundance of nine transcripts (i.e. MOGAT1, PFKB2, LYZ2, SVS8, UHRF1, PTGES, AGPAT4, DGKA and HGPD) of 53 tested in the endometrial tissue differed between heifers with a viable or retarded embryo. Both LYZ2 and UHRF1 are associated with regulation of the immune system; PFKFB2 is a mediator in glycolysis; MOGAT, AGPAT4 and DGKA belong to the triglyceride synthesis pathway; and PTGES and HGPD belong to the prostaglandin pathway. Both these metabolic pathways are important for early embryonic development. In conclusion, retarded embryo development in the present study was not related to serum progesterone, IGF-I, insulin or urea concentrations, nor to uterine pH at the time of death. However, altered expression of genes involved in the prostaglandin and triglyceride pathways, as well as two genes that are closely associated with the regulation of immunity, in the endometrium may indicate a uterine component in the retardation of embryo development in these beef heifers.

Effect of progesterone supplementation in the first week post conception on embryo survival in beef heifers.

Progesterone is essential for establishment and maintenance of pregnancy in mammals. The objective of this study was to examine the effect of elevating progesterone during the different physiological stages of early embryo development on embryo survival. Estrus was synchronized in cross-bred beef heifers (n=197, approximately 2-years old) and they were inseminated 12-18h after estrus onset (=Day 0). Inseminated heifers were randomly assigned to 1 of 3 treatments: (1) Control, n=69; (2) progesterone supplementation using a Controlled Internal Drug Release Device (CIDR) from Day 3 to 6.5, n=64; or (3) progesterone supplementation using a CIDR from Day 4.5 to 8, n=64. Body condition (BCS) and locomotion scores (scale of 1-5) were recorded for all animals. Animals with a locomotion score >/=4 (very lame) were excluded. Embryo survival rate was determined at slaughter on Day 25. Conceptus length and weight were recorded and the corpus luteum (CL) of all pregnant animals was dissected and weighed. Supplementation with exogenous progesterone increased (P<0.05) peripheral progesterone concentrations, but did not affect embryo survival rate compared with controls. Mean CL weight, conceptus length and conceptus weight were not different between treatments. There was a positive relationship (P<0.04) between the increase in progesterone concentrations from Days 3 to 6.5 and embryo survival rate in treated heifers and a similar trend existed between the increase from Days 4.5 to 8 (P<0.06). There was also a positive relationship (P<0.05) between the progesterone concentration on Day 6.5 and the embryo survival rate in treated heifers. A direct correlation was seen between locomotion score and embryo survival rate, with higher (P<0.05) early embryo survival rates in heifers with a lower locomotion score. In conclusion, supplementation with progesterone at different stages of early embryo development increased peripheral progesterone concentration and resulted in a positive association between changes in progesterone concentration during the early luteal phase and embryo survival rate. Supplementation with progesterone had no effect on either CL weight or conceptus size in pregnant animals. Lameness had a significant negative effect on early embryo survival.

Hormonal composition of follicular fluid from abnormal follicular structures in mares.

The objective was to characterise the hormonal composition of follicular fluid from mares with distinct anovulatory-cystic follicles. Follicular fluid was aspirated from six mares that presented with cystic follicles and from pre-ovulatory follicles of five normal mares (controls). Differences in progesterone, oestradiol, testosterone, IGF-I and IGF binding were analysed using Fisher's exact test. There were greater (P < 0.03) follicular fluid oestradiol concentrations in normal follicles and the testosterone concentration of the cystic fluid was greater (P < 0.05) than that of the normal fluid. There also was a greater (P < 0.03) percentage of IGF-I binding and lower (P < 0.02) IGF-I concentrations in the fluid collected from the cystic structures compared with the fluid from normal follicles. Despite the limited number of animals, the fact that fluid aspirated from cystic follicles had higher testosterone and lower oestradiol concentrations could be of diagnostic value when a practitioner wants to distinguish between a cystic and non-cystic persistent follicle. The research reported here also indicates a likely role for the IGF system in the pathogenesis of the development and maintenance of anovulatory follicular structures in mare ovaries.

Global proteomic characterization of uterine histotroph recovered from beef heifers yielding good quality and degenerate day 7 embryos.

The objective was to analyze the proteomic composition of uterine flushes collected from beef heifers on day 7 after insemination. Estrus was synchronized in crossbred beef heifers by using a protocol with a controlled intravaginal drug releasing device. Heifers detected in standing estrus (within 24-48 h after removal of controlled intravaginal drug releasing device) were inseminated (estrus = day 0) with frozen-thawed semen from a single ejaculate of a bull with proven fertility. Heifers from which an embryo was recovered (after slaughter on day 7) were classified as either having a viable embryo (morula/blastocyst stage) or a degenerate embryo (arrested at the 2- to 16-cell stage). The overall recovery rate (viable and degenerate combined) was 64%. Global liquid chromatography coupled to tandem mass spectrometry proteomic analysis of the histotroph collected identified 40 high-confidence proteins present on day 7; 26 proteins in the viable group, 10 in the degenerate group, and 4 shared between both groups. Five proteins (platelet-activating factor acetylhydrolase IB subunit γ [PAFAH1B3], tubulin α-1D chain, tubulin ß-4A chain, cytochrome C, and dihydropyrimidinase-related protein-2) were unique or more abundant in the histotroph collected from animals with a viable embryo, and 1 protein (S100-A4) was more abundant in the histotroph collected from animals with a degenerate embryo. Of interest, PAFAH1B3, detected only in histotroph from the group yielding viable embryos, belongs to the group of platelet-activating factors that are known to be important for the development of the pre-implantation embryo in other species. To our knowledge this is the first report of PAFAH1B3 in relation to bovine early embryonic development.

Interovulatory intervals in mares receiving deslorelin implants in Ireland (2009 to 2010).

Deslorelin acetate implants, recently licensed in Ireland and the UK for ovulation induction in mares, have been associated with prolonged interovulatory intervals in USA studies, leading to the practice of removing implants postovulation. Trial data in Australia indicate a less pronounced effect on interovulatory intervals, suggesting possible geographical variation. Objectives of the current study were to assess the effect of deslorelin implants, with and without removal on oestrous cycle length in Irish- and UK-based Thoroughbred broodmares. Data were collected retrospectively from 88 oestrous cycles. A statistically significant difference (P=0.02) was found between interovulatory intervals in mares in which the deslorelin implant was not removed, compared with administration and removal of the implant or the use of human chorionic gonadotrophin. The results suggest that implant removal when possible is advisable. The delay in subsequent ovulations was less marked than that reported in some studies from the USA. This information is useful in deciding when to schedule subsequent breeding for mares which received a deslorelin implant during the previous oestrous period and provides evidence to counter-concerns that mares treated with deslorelin implants may experience a long delay in return to oestrus if the implant is not removed.

Oestrous cycles in Bos taurus cattle.

The oestrous cycle in cattle lasts for 18-24 days. It consists of a luteal phase (14-18 days) and a follicular phase (4-6 days). During the cycle there are generally two (dairy cows) or three (heifers and beef cows) waves of ovarian follicle growth. Each wave of follicle growth consists of a period of emergence of a cohort of follicles, selection of a dominant follicle and either atresia or ovulation of the dominant follicle. These waves of follicle growth, initially established during the early pre-pubertal period of development occur throughout the entire cycle, with only the dominant follicle (DF) of the final wave coinciding with the follicular phase that undergoes final maturation and ovulation. Ovarian functions (follicle growth, ovulation, luteinisation and luteolysis) are regulated by the endocrine hormones of the hypothalamus (gonadotrophin-releasing hormone), anterior pituitary (follicle-stimulating hormone and luteinising hormone), ovaries (progesterone, oestradiol and inhibins) and the uterus (prostaglandin F2α). In postpartum cows resumption of regular oestrous cycles (in addition to uterine involution) is fundamental for re-establishment of pregnancy.

The effect of body condition at calving and supplementation with Saccharomyces cerevisiae on energy status and some reproductive parameters in early lactation dairy cows.

Improving the energy status of dairy cows during the early post-partum (PP) period by adding a safe dietary supplement such as live yeast culture (YS) may have a positive effect on reproductive function. The objective was to examine potential benefits of YS supplementation on PP energy status and fertility indices of dairy cows managed to have low or high body condition score (BCS, 1-5 scale) at calving. Forty (10 primiparous and 30 multiparous) Holstein/Friesian dairy cows were blocked by yield, parity, BCS, and predicted calving date. Within each block, cows were randomly allocated to a 2 x 2 factorial arrangement of treatments which were: BCS at calving (low < or =3.5 or high > or =3.75; n=20) and YS supplementation (2.5g/cow/day for pre-calving and 10g/cow/day for post-calving x 10(8) CFU of Saccharomyces cerevisiae/g) (supplemented or control; n=20). Daily milk yield was recorded and weekly milk composition, BCS and BW were assessed from calving to week 10 PP. Forage (100% grass silage pre-calving; 50% grass silage, 50% maize silage post-calving; ad libitum) intake was recorded individually. Concentrate (2kg of pre-calver nuts+/-YS for pre-calving and 8 kg of lactating nuts+/-YS for post-calving) feeding was controlled individually. Estimated energy balance PP was calculated on a weekly basis individually as the difference between the net energy (NE) intake and the sum of NE for maintenance and milk production. Insulin and IGF-I concentrations were determined on days 14 and 7 pre-calving and 1, 5, 15, 25 and 35 post-calving. Daily ovarian ultrasonography was performed from day 10 PP to monitor the size and development of the first dominant follicle (>10mm in diameter with absence of other large growing follicles), first ovulatory follicle and days to first ovulation PP. Pre-ovulatory peak of serum oestradiol concentration was determined during the 2 days before ovulation day. Cows with high BCS (over-conditioned) at calving ingested less NE, produced more milk NE output, and consequently had a significantly (P<0.05) exacerbated negative energy balance in comparison with low BCS cows (moderately conditioned) during early lactation. Higher (P<0.05) insulin concentrations and a tendency for higher (P=0.06) pre-ovulatory peak oestradiol concentrations in low BCS group were detected in the early PP period. Supplementing the diet with YS had no effect (P>0.10) on NE intake, NE milk output or energy balance. On the other hand it increased (P<0.01) insulin concentration and tended to increase (P=0.07) pre-ovulatory peak oestradiol concentrations and the size of first ovulatory follicle (P=0.09) early PP. Feeding YS had no effect on energy status of lactating dairy cows with high or low BCS at calving, whilst it improved serum insulin concentration, pre-ovulatory peak of oestradiol and the size of first ovulatory follicle in the early PP period. These observed effects of YS supplementation require to be substantiated with further research.

Evaluation of models to induce low progesterone during the early luteal phase in cattle.

Two experiments were designed to evaluate models for generation of low circulating progesterone concentrations during early pregnancy in cattle. In Experiment 1, 17 crossbred heifers (Bos taurus) were assigned to either prostaglandin F(2alpha) (PGF(2alpha)) administration on Days 3, 3.5, and 4 (PG3; n=9) or to control (n=8). Blood samples were collected from heifers from Days 1 to 9 for progesterone assay. Progesterone concentrations were decreased (P<0.03) between 18 and 48h after first PGF(2alpha) treatment in heifers assigned to PG3 compared with that of controls. In Experiment 2, 39 crossbred heifers detected in estrus were inseminated (Day 0) and assigned to either (1) PGF(2alpha) administration on Days 3, 3.5, and 4 (PG3; n=10), (2) PGF(2alpha) administration on Days 3, 3.5, 4, and 4.5 (PG4; n=10), (3) Progesterone Releasing Intravaginal Device (PRID) insertion on Day 4.5 with PGF(2alpha) administration on Days 5 and 6 (PRID+PGF(2alpha); n=10), or (4) control (n=9). Blood samples were collected daily until Day 15, and conceptus survival rate was determined at slaughter on Day 16. Progesterone concentrations during the sampling period in the PG3 and PG4 groups did not differ but were less than that of controls (P<0.01). After an initial peak, progesterone concentrations in the PRID+PGF(2alpha) group were similar to that of controls. More heifers in the PG4 group (6 of 10) had complete luteal regression than did those in the PG3 group (3 of 10). Conceptus survival rate on Day 16 did not differ between groups. There was a significant correlation between progesterone concentration on Days 5 and 6 and conceptus size on Day 16. In summary, treatment with PGF(2alpha) on Days 3, 3.5, and 4 postestrus appeared to provide the best model to induce reduced circulating progesterone concentrations during the early luteal phase in cattle.