Short communication: Validation of a novel milk progesterone-based tool to monitor luteolysis in dairy cows using cost-effective,on-farm measured data

The progesterone (P4) monitoring algorithm using synergistic control (PMASC) uses luteal dynamics to identify fertility events in dairy cows. This algorithm employs a combination of mathematical functions describing the increasing and decreasing P4 concentrations during the development and regression of the corpus luteum and a statistical control chart that allows identification of luteolysis. The mathematical model combines sigmoidal functions from which the cycle characteristics can be calculated. Both the moment at which luteolysis is detected and confirmed by PMASC, as well as the model features themselves, can be used to inform the farmer on the fertility status of the cows.     

A novel system for on-farm fertility monitoring based on milk progesterone

Timely identification of a cow's reproduction status is essential to minimize fertility-related losses on dairy farms. This includes optimal estrus detection, pregnancy diagnosis, and the timely recognition of early embryonic death and ovarian problems. On-farm milk progesterone (P4) analysis can indicate all of these fertility events simultaneously. However, milk P4 measurements are subject to a large variability both in terms of measurement errors and absolute values between cycles. The objective of this paper is to present a newly developed methodology for detecting luteolysis preceding estrus and give an indication of its on-farm use. The innovative monitoring system presented is based on milk P4 using the principles of synergistic control. Instead of using filtering techniques and fixed thresholds, the present system employs an individually on-line updated model to describe the P4 profile, combined with a statistical process control chart to identify the cow's fertility status. The inputs for the latter are the residuals of the on-line updated model, corrected for the concentration-dependent variability that is typical for milk P4 measurements. To show its possible use, the system was validated on the P4 profiles of 38 dairy cows. The positive predictive value for luteolysis followed by estrus was 100%, meaning that the monitoring system picked up all estrous periods identified by the experts. Pregnancy or embryonic mortality was characterized by the absence or detection of luteolysis following an insemination, respectively. For 13 cows, no luteolysis was detected by the system within the 25 to 32 d after insemination, indicating pregnancy, which was confirmed later by rectal palpation. It was also shown that the system is able to cope with deviating P4 profiles having prolonged follicular or luteal phases, which may suggest the occurrence of cysts. Future research is recommended for optimizing sampling frequency, predicting the optimal insemination window, and establishing rules to detect problems based on deviating P4 patterns.     

Evaluation and characterization of estrus alerts and behavioral parameters generated by an ear-attached accelerometer-based system for automated detection of estrus

Our objectives were to evaluate the performance of an ear-attached automated estrus detection (AED) system (Smartbow; Zoetis) that monitored physical activity and rumination time, and to characterize AED system estrus alert features (i.e., timing and duration). Lactating Holstein cows (n = 216) commenced a protocol for the synchronization of estrus at 50 ± 3 DIM or 18 ± 3 d after artificial insemination. For 7 d after induction of luteolysis with PGF_2α (d 0), we used visual observation of estrous behavior (30 min, 2 times per day) and data from an automated mounting behavior monitoring system based on a pressure-activated tail-head sensor (HeatWatch; Cowchips LLC) as a reference test (RTE) to detect behavioral estrus. Concomitantly, estrus alerts and their features were collected from the AED system. Progesterone levels confirmed luteal regression, and transrectal ultrasonography confirmed the occurrence and timing of ovulation. Performance metrics for the AED system were estimated with PROC FREQ in SAS, using the RTE or ovulation only as a reference. Performance was also estimated after the removal of cows with a discrepancy between the RTE and ovulation. Continuous outcomes with or without repeated measurements were evaluated by ANOVA using PROC MIXED in SAS. Based on the RTE, 86.6% (n = 187) of the cows presented estrus and ovulated; 1.4% (n = 3) presented estrus and did not ovulate; 6.4% (n = 14) did not present estrus but ovulated; and 5.6% (n = 12) did not present estrus or ovulation. We found no difference in the proportion of cows detected in estrus and with ovulation for the AED system (83.4%) and the RTE (86.6%). Compared with estrus events as detected by the RTE, sensitivity for the AED was 91.6% (95% CI: 87.6–95.5) and specificity was 69.2% (95% CI: 51.5–87.0). Using ovulation as reference, sensitivity was 89.6% (95% CI: 85.3–93.8) and specificity was 86.7% (95% CI 69.5–100). For all cows with agreement between the RTE and ovulation, sensitivity was 92.5% (95% CI: 88.7–96.3) and specificity was 91.7% (95% CI: 76.0–100). The mean (±SD) interval from induction of luteolysis to estrus alerts, estrus alert duration, and the onset of estrus alerts to ovulation interval were 72.2 ± 18.1, 13.5 ± 3.8, and 23.8 ± 7.1 h, respectively. We concluded that an ear-attached AED system that monitored physical activity and rumination time was effective at detecting cows in estrus and generated few false positive alerts when accounting for ovulation, cow physiological limitations, and the limitations of the RTE.     

Effects of d-cloprostenol dose and corpus luteum age on ovulation, luteal function, and morphology in nonlactating dairy cows with early corpora lutea

Luteolysis is a key event in cattle reproduction. A standard dose of exogenous PGF(2α) will induce full luteolysis in the majority of cows with a matured corpus luteum (CL). However, this will not occur in cows with a CL <5d old. To date, it is not known whether a larger dose will have a more potent luteolytic effect in cows during early diestrus. The objective of this study was to characterize the effect of 2 doses of d-cloprostenol (150 and 300 μg) on the progesterone concentration, luteal diameter, and ovulation rate in nonlactating dairy cattle 96 to 132 h postovulation. Twenty nonlactating dairy cows were included in the study. Each cow received 2 treatments of d-cloprostenol in 2 consecutive cycles: a standard dose of 150 μg and a double dose of 300 μg. The cows were allocated randomly to 1 of 4 groups (5 cows in each group) according to the age of the CL at the time of treatment: 96, 108, 120, and 132 h. The exact time of ovulation was known within 12h, because of twice per day ultrasound examination. The CL diameter and progesterone concentration were measured before treatment (d 0) and 2 and 4d after treatment. Within each CL age group, the effect of d-cloprostenol dose on luteolysis was determined. More cows treated with double dose tended to have full luteolysis compared with the standard dose (8/10 vs. 4/10, respectively). This effect was only apparent in cows with CL of 120 and 132 h but not in earlier CL. The interval from treatment to ovulation was shorter (3.3 ± 0.1d) in cows treated with a double dose than in cows treated with the standard dose (4.5 ± 0.4d).     

Cost-benefit evaluation of on-farm milk progesterone testing to monitor return to cyclicity and to classify ovarian cysts.

The effect of using on-farm milk progesterone testing to monitor return to cyclicity after parturition and to classify correctly and to treat cystic cows was evaluated using modeling and simulation. The test was evaluated assuming low and high accuracies of progesterone measurement, first breeding policy of 40 d, average estrus detection rate of 55%, and an average fertility rate of 55%. Three testing schemes (starting milk progesterone testing on d 30, 40, or 50 after parturition) were compared against a control to evaluate the effect of monitoring return to cyclicity on cows' reproductive and economic performance. For this purpose, the use of the test was an economically justifiable management intervention. Starting to test on d 30 after parturition was the most effective scheme, reducing days open by 18 d, replacement rate by 2.4%, and increasing net return per cow per year by $11. Starting to test on d 50 postpartum was not economically justifiable. Accuracy of the test, within the range used in this study, was unimportant. Testing was most profitable in herds with low fertility and low efficiency of estrus detection. The use of the test to classify follicular and luteal cysts and to select the appropriate therapy was not economically justifiable because of the low proportion of cystic cows and the high variation in response to therapy.     

Luteolysis,progesterone, and pregnancy per insemination after modifying the standard 7-day Ovsynch program in Holstein-Friesian and Holstein cows

Two experiments were conducted with Holstein-Friesian cows in the Republic of North Macedonia and with Holstein cows in Kansas. We hypothesized that 1 dose of PGF_2α administered on d 8 (Ov-8×1) instead of d 7 (Ov-7×1) in an Ovsynch program [GnRH-1 (d 0)–7 d–PGF_2α–56 h–GnRH-2–16 h–timed artificial insemination (AI)] would increase the proportion of cows with complete luteolysis compared with controls receiving a single dose on d 7. Cows were treated with Ov-7×1 or with Ov-8×1 in experiment 1 (n = 347), using only a single dose of PGF_2α. In experiment 2 (n = 452), a third treatment was added (Ov-7×2), in which a second dose of PGF_2α was administered on d 8. Progesterone was measured in blood samples collected before the first or only PGF_2α administration and 72 h later before insemination. Complete luteolysis was defined as having occurred when progesterone was ≥1 ng/mL before PGF_2α and ≤0.3 ng/mL 72 h later (time of AI). Follicles and luteal structures were mapped before GnRH-1 and PGF_2α administrations. The results of experiment 1 demonstrated a greater percentage of multiparous cows in OV-8×1 having complete luteal regression compared with multiparous Ov-7×1 cows, whereas treatments were equally effective in primiparous cows, as reflected in the concentrations of progesterone before AI. Furthermore, pregnancy per AI did not differ between treatments. Results in experiment 2 revealed that 99.3% of cows in the Ov-7×2 treatment receiving the second dose of PGF_2α had complete luteolysis, regardless of parity, compared with significantly fewer cows in the Ov-7×1 and Ov-8×1 treatments (91.2 and 90.6%, respectively). Neither concentrations of progesterone, which averaged <0.4 ng/mL at AI, nor pregnancy per AI differed among the 3 treatments. In both experiments, when status of luteal function before PGF_2α treatment was examined [cows with no corpus luteum (CL) before GnRH-1 but which had formed a new CL in response to ovulation after GnRH-1; cows with an older CL (the same CL that was detected before GnRH-1); or cows with both a new and an older CL], treatments did not differ in causing complete luteolysis. Furthermore, complete luteolysis in experiment 2 did not differ regardless of whether cows had 1, 2, or 3 or more CL before PGF_2α administration. Pregnancy per AI did not differ among treatments, indicating that any of the 3 treatments might produce similar pregnancy outcomes with the flexibility of applying either of the 7- or the 8-d treatments.     

Genetic parameters for atypical reproductive patterns in dairy cows estimated from in-line milk progesterone profiles

Our aim was to estimate genetic parameters of atypical reproductive patterns and estimate their genetic correlation with milk production and classical fertility traits for commercial dairy cows. In contrast with classical fertility traits, atypical reproductive patterns based on in-line milk progesterone profiles might have higher heritability and lower genetic correlation with milk production. We had in-line milk progesterone profiles available for 12,046 cycles in 4,170 lactations of 2,589 primiparous and multiparous cows (mainly Holstein Friesian) from 14 herds. Based on progesterone profiles, 5 types of atypical reproductive patterns in a lactation were defined: delayed ovulation types I and II, persistent corpus luteum types I and II, and late embryo mortality. These atypical patterns were detected in 14% (persistent corpus luteum type II) to 21% (persistent corpus luteum type I) of lactations. In 47% of lactations, at least 1 atypical pattern was detected. Threshold model heritabilities for atypical reproduction patterns ranged between 0.03 and 0.14 and for most traits were slightly higher compared with classical fertility traits. The genetic correlation between milk yield and calving interval was 0.56, whereas genetic correlations between milk yield and atypical reproductive patterns ranged between ?0.02 and 0.33. Although most of these correlations between milk yield and atypical reproductive patterns are still unfavorable, they are lower compared with the correlations between classical fertility traits and milk yield. Therefore selection against atypical reproductive patterns may relax some constraints in current dairy breeding programs, to enhance genetic progress in both fertility and milk yield at a steady pace. However, as long as the target trait for fertility is calving interval, atypical reproductive patterns will not add additional value to the breeding goal in the near future due to the low number of available records.     

Atypical progesterone profiles and fertility in Swedish dairy cows

The incidence of normal and atypical progesterone profiles in Swedish dairy cows was studied. Data were collected from an experimental herd over 15 yr, and included 1,049 postpartum periods from 183 Swedish Holstein and 326 Swedish Red and White dairy cows. Milk progesterone samples were taken twice weekly until initiation of cyclical ovarian activity and less frequently thereafter. Progesterone profiles were 1) normal profile: first rise in milk progesterone above the threshold value before d 56 postpartum, followed by regular cyclical ovarian activity (70.4%); 2) delayed onset of cyclical ovarian activity: low milk progesterone the first 56 d postpartum (15.6%); 3) cessation of cyclical ovarian activity: ovarian activity resumed within 56 d postpartum, but ceased for a period of 14 d or more (6.6%); and 4) prolonged luteal phase: ovarian activity resumed within 56 d postpartum, but milk progesterone remained elevated in the nonpregnant cow for a period of 20 d or more (7.3%). Swedish Holsteins had 1.5 times higher risk of atypical profile than Swedish Red and Whites. Risk of atypical profiles was 0.5 and 0.7 times lower for older cows compared with first-parity cows; 2.3 times higher for cows in tie-stalls compared with those in loose housing; 2.6 times higher for cows calving during winter compared with summer; 0.5 times lower for cows in earlier (1994-1999) calving-year groups compared with the most recent (2000-2002); 2.5 times higher for cows with planned extended calving interval compared with conventional calving interval; and 2.2 times higher for an atypical profile in previous lactation compared with a normal profile. Cows with atypical profiles had a 15-d increase in interval from calving to first artificial insemination and an 18-d increase in interval from calving to conception. Progesterone samples taken within the first 60 d postpartum were used to calculate the percentage of samples above the threshold value of luteal activity. This measure had a significantly different mean in profiles and can be used to separate delayed onset of cyclical ovarian activity profiles and prolonged luteal phase profiles from normal. Thereby, it may be a more effective tool than measurements based only on the onset of ovarian cyclical activity in genetic evaluation of early postpartum fertility in dairy cows.     

Candidate mechanisms underlying atypical progesterone profiles as deduced from parameter perturbations in a mathematical model of the bovine estrous cycle

The complex interplay of physiological factors that underlies fertility in dairy cows was investigated using a mechanistic mathematical model of the dynamics of the bovine estrous cycle. The model simulates the processes of follicle and corpus luteum development and its relations with key hormones that interact to control these processes. Several factors may perturb the regular oscillatory behavior of a normal estrous cycle, and such perturbations are likely the effect of simultaneous changes in multiple parameters. The objective of this paper was to investigate how multiple parameter perturbation changes the behavior of the estrous cycle model, so as to identify biological mechanisms that could play a role in the development of cystic ovaries. Cystic ovaries are a common reason for reproductive failure in dairy cows, but much about the causes of this disorder remains unknown. We investigated in which region of the parameter space the model predicts a normal cycle, and when a progesterone pattern occurred with delayed ovulation (indicating a cystic follicle) or delayed luteolysis (indicating a persistent corpus luteum). Perturbation of the initial values for all parameters simultaneously showed 2 specific parameter configurations leading to delayed ovulation or delayed luteolysis immediately. The most important parameter changes in these 2 configurations involve the regulation of corpus luteum functioning, luteolytic signals, and GnRH synthesis, suggesting that these mechanisms are likely involved in the development of cystic ovaries. In the multidimensional parameter space, areas exist in which the parameter configurations resulted in normal cycles. These areas may be separated by areas in which irregular cycle patterns occurred. These irregular patterns thus mark the transition from one stable (normal) situation to another. Interestingly, within a series, there were some cycles with delayed ovulation and some with delayed luteolysis in these patterns. This could represent a situation of resumption of normal cyclicity (e.g., after parturition). In conclusion, the method of parameter perturbation used in the present study is an effective tool to find parameter configurations that lead to progesterone profiles associated with delayed ovulation and delayed luteolysis. Thereby, the model helps to generate hypotheses regarding the underlying cause of the development of cystic ovaries, which could be investigated in future experiments.     

Replacing the first gonadotropin-releasing hormone treatment in an Ovsynch protocol with human chorionic gonadotropin decreased pregnancies per artificial insemination in lactating dairy cows

Our objective was to compare the effect of treatment with GnRH at the first treatment (G1) of the Breeding-Ovsynch portion of a Double-Ovsynch (DO) protocol with human chorionic gonadotropin (hCG) on pregnancies per artificial insemination (P/AI) in lactating dairy cows. In experiment 1, lactating dairy cows (n = 1,932) submitted to a DO protocol for first timed artificial insemination (TAI) on 2 commercial dairy farms were blocked by parity (primiparous vs. multiparous) and were randomly assigned to receive 100 µg of GnRH versus 2,500 IU of hCG at G1. Overall, P/AI 39 d after TAI for cows inseminated with sexed dairy semen was greater for cows treated with GnRH than for cows treated with hCG within each parity (primiparous: 42.6% vs. 38.2%; multiparous: 39.4% vs. 30.3%). Similarly, P/AI 39 d after TAI for multiparous cows inseminated with conventional beef semen tended to be greater for cows treated with GnRH than for cows treated with hCG (41.1% vs. 34.3%). In experiment 2, lactating Holstein cows (n = 43) were blocked by parity and were randomly assigned to the treatment protocols described for experiment 1. Ovaries were evaluated with transrectal ultrasonography immediately before treatment and 24, 28, 32, 36, and 40 h after treatment to assess time from treatment to ovulation, and blood samples were collected immediately before G1, at the first PGF_2α treatment, 8 and 16 h later, at the second PGF_2α treatment, 8 and 16 h later, at the second GnRH (G2) treatment, and at TAI to compare luteolysis based on serum progesterone (P4) concentrations. Although mean (± standard error of the mean) time from treatment to ovulation was approximately 2 h greater for cows treated with hCG than for cows treated with GnRH (33.7 ± 0.6 vs. 31.5 ± 0.6 h), P4 concentrations during luteolysis and the proportion of cows with complete luteolysis (P4 <0.4 ng/mL at G2) did not differ between treatments. We conclude that replacing 100 µg of GnRH with 2,500 IU of hCG at G1 of a DO protocol decreased fertility to TAI in lactating dairy cows but did not affect the rate or completeness of luteolysis despite the increased interval from treatment to ovulation.     

Assessment of an accelerometer system for detection of estrus and treatment with gonadotropin-releasing hormone at the time of insemination in lactating dairy cows

Two experiments were conducted to evaluate an accelerometer system (Heatime; SCR Engineers Ltd., Netanya, Israel) to manage reproduction in lactating dairy cows. In experiment 1, lactating Holstein cows (n = 112) were fitted with an accelerometer system and were treated with GnRH followed 7 d later by PGF_2α to synchronize estrus. A total of 89 cows that had a follicle >10 mm in diameter and a functional corpus luteum at the PGF_2α injection that regressed by 48 h after induction of luteolysis were included in the analysis. Overall, 71% of cows were detected in estrus by the accelerometer system and 95% of cows showing estrus ovulated within 7 d after induction of luteolysis. Of the cows not detected in estrus by the accelerometer system, 35% ovulated within 7 d after induction of luteolysis. Duration of estrus activity (mean ± SD) was 16.1 ± 4.7 h and was neither affected by parity nor milk production. Intervals (means ± SD) from induction of luteolysis, onset of activity, peak raw activity, and peak weighted activity to ovulation was 82.2 ± 9.5, 28.7 ± 8.1, 20.4 ± 7.8, and 16.4 ± 7.4 h, respectively, and the interval from AI to ovulation was 7.9 ± 8.7 h, but ranged from −12 to 26 h. In experiment 2, cows were assigned randomly to receive an intramuscular injection of GnRH at artificial insemination (AI) after detection of estrus by the accelerometer system or receive no treatment (control). Nine hundred seventy-nine AI services from 461 cows were analyzed. Treatment with GnRH at AI did not affect fertility at 35 or 65 d after AI, and no interaction was detected between treatment and season or treatment and AI number. Overall, two-thirds of the cows that were considered properly synchronized were inseminated based on the accelerometer system and ovulated after AI. The remaining cows either were not inseminated because they were not detected in estrus or would not have had a chance to conceive to AI because they failed to ovulate after estrus. Furthermore, mean time of AI in relation to ovulation determined by the accelerometer system was acceptable for most of the cows that displayed estrus; however, variability in the duration of estrus and timing of AI in relation to ovulation could lead to poor fertility in some cows. For lactating dairy cows detected in estrus by the accelerometer system, treatment with GnRH at the time of AI without reference to the onset of estrus did not increase fertility.     

Follicular wave of the ovulatory follicle and not cyclic status influences fertility of dairy cows

Two experiments evaluated the influence of follicular wave at artificial insemination (AI) on fertility of dairy cows. In experiment 1, data from 5,607 lactating cows enrolled in estrous and ovulation synchronization programs for AI were evaluated. Cows’ blood was analyzed for progesterone 7 to 14 d apart, with the second sample collected on the day of the first GnRH (GnRH1) of the synchronization protocol. Cows were classified as cyclic if progesterone was ≥1 ng/mL in at least 1 of the 2 samples and as anovular if both samples were <1 ng/mL. Cyclic cows were categorized as low (CLOW; < 1 ng/mL) or high (CHIGH; ≥ 1 ng/mL) progesterone on the day of GnRH1, which would result in ovulation of the dominant follicle of the first (FW) and second (SW) follicular waves, respectively, at AI. Pregnancy per AI (P/AI) was determined 30 and 53 d after AI. In experiment 2, 220 cyclic Holstein cows received 2 injections of PGF_2α administered 14 d apart. The Ovsynch protocol (d 0 GnRH, d 7 PGF_2α, d 9 GnRH, d 9.5 timed AI) was initiated either 3 or 10 d after the second PGF_2α of the presynchronization to result in insemination to the FW or SW dominant follicles. Blood was analyzed for progesterone and ovaries were scanned to determine ovulatory responses and follicle diameter. Pregnancy was determined on d 32 and 67 after timed AI. In experiment 1, P/AI on d 30 was greater for CHIGH cows than for CLOW and anovular cows (43.0, 31.3, and 29.7%, respectively), but because of pregnancy loss, P/AI on d 53 was lowest for anovular cows. Proportions of cows with short reinsemination intervals differed among groups and were 7.1, 15.7, and 11.9% for CHIGH, CLOW, and anovular cows, respectively. Pregnancy loss was greater for anovular cows than for CLOW cows (15.0 vs. 10.0%) and was intermediate for CHIGH cows (13.5%). In experiment 2, 9.8 and 97.2% of the FW and SW cows, respectively, had progesterone ≥1 ng/mL at GnRH1. Concentrations of progesterone at the GnRH1 and PGF_2α injections of the Ovsynch protocol were greater for SW cows than FW cows. Pregnancy per AI was greater for SW cows than for FW cows (41.7 vs. 30.4%) despite less ovulation to GnRH1 in SW cows than in FW cows (78.7 vs. 88.4%). Collectively, these data indicate that follicular wave of the ovulatory follicle and not cyclic status caused the greatest reduction in P/AI in dairy cows. Whether the culprit is the follicle itself or the hormonal milieu characteristic of the first follicular wave and the early stage of the estrous cycle remains to be elucidated. Synchronization programs that induced ovulation of the FW follicle at AI reduced P/AI in lactating dairy cows, and ovulation of the FW follicle, or development of the ovulatory follicle under low progesterone concentrations, or both, might be mechanisms for reduced fertility in anovular cows.     

Characterization of progesterone profiles in fall-calving Norwegian Red cows

Progesterone profiles in Norwegian Red cows were categorized, and associations between the occurrence of irregularities in the profiles and the commencement of luteal activity were investigated. The cows were managed in 3 feeding trials from 1994 to 2001 and from 2005 to 2008 at the Norwegian University of Life Sciences. The cows were followed from calving, and the milk samples collected represented 502 lactations from 302 cows. Milk samples for progesterone analysis were taken 3 times weekly from 1994 throughout 1998 and from 2005 to 2008 and 2 times weekly from 1999 to 2001. Commencement of luteal activity was defined as the first day of 2 consecutive measurements of progesterone concentration ≥3 ng/mL not earlier than 10 d after calving. Delayed ovulation type I was defined as consistently low progesterone concentration, <3 ng/mL for ≥50 d postpartum. Delayed ovulation type II was defined as prolonged interluteal interval with milk progesterone measurements <3 ng/mL for ≥12 d between 2 luteal phases. Persistent corpus luteum (PCL) type I was defined as delayed luteolysis with milk progesterone ≥3 ng/mL for ≥19 d during the first estrous cycle postpartum. Persistent corpus luteum type II was defined as delayed luteolysis with milk progesterone ≥3 ng/mL for ≥19 d during subsequent estrous cycles before first artificial insemination. Delayed ovulation type I was present in 14.7%, delayed ovulation type II in 2.8%, PCL type I in 6.7%, and PCL type II in 3.3% of the profiles. Commencement of luteal activity was related to milk yield, parity, PCL type I, and the summated occurrence of PCL type I and II. The least squares means for the interval to commencement of luteal activity were 24.2 d when PCL type I and II were present and 29.5 d when PCL type I and II were absent. The likelihood of pregnancy to first service was not affected in cows with a history of PCL when artificial insemination was carried out at progesterone concentrations <3 ng/mL (i.e., during estrus); however, cows that had experienced PCL were more likely to be inseminated during a luteal phase. The occurrence of delayed ovulation and PCL in Norwegian Red cows was less than that reported in most other dairy populations.     

Effects of supplemental progesterone after artificial insemination on expression of interferon-stimulated genes and fertility in dairy cows

The objectives of the current study were to evaluate the effects of supplemental progesterone after artificial insemination (AI) on expression of IFN-stimulated genes (ISG) in blood leukocytes and fertility in lactating dairy cows. Weekly cohorts of Holstein cows were blocked by parity (575 primiparous and 923 multiparous) and method of insemination (timed AI or AI on estrus) and allocated randomly within each block to untreated controls, a controlled internal drug release (CIDR) containing 1.38 g of progesterone from d 4 to 18 after AI (CIDR4), or a CIDR on d 4 and another on d 7 after AI and both removed on d 18 (CIDR4+7). Blood was sampled to quantify progesterone concentrations in plasma and mRNA expression in leukocytes for the ubiquitin-like IFN-stimulated gene 15-kDa protein (ISG15) and receptor transporter protein-4 (RTP4) genes. Pregnancy was diagnosed on d 34 ± 3 and 62 ± 3 after AI. Treatment increased progesterone concentrations between d 5 and 18 after AI in a dose-dependent manner (control = 3.42, CIDR4 = 4.97, and CIDR4+7 = 5.46 ng/mL). Cows supplemented with progesterone tended to have increased luteolysis by d 19 after AI (control = 17.2; CIDR4 = 29.1; CIDR4+7 = 30.2%), which resulted in a shorter AI interval for those reinseminated after study d 18. Pregnancy upregulated expression of ISG in leukocytes on d 19 of gestation, but supplementing progesterone did not increase mRNA abundance for ISG15 and RTP4 on d 16 after insemination and tended to reduce mRNA expression on d 19 after AI. For RTP4 on d 19, the negative effect of supplemental progesterone was observed only in the nonpregnant cows. No overall effect of treatment was observed on pregnancy per AI on d 62 after insemination and averaged 28.6, 32.7, and 29.5% for control, CIDR4, and CIDR4+7, respectively. Interestingly, an interaction between level of supplemental progesterone and method of AI was observed for pregnancy per AI. For cows receiving exogenous progesterone, the lower supplementation with CIDR4 increased pregnancy per AI on d 62 in cows inseminated following timed AI (CIDR4 = 39.2; CIDR4+7 = 27.5%); in those inseminated following detection of estrus, however, the use of a second insert on d 7 resulted in greater pregnancy per AI (CIDR4 = 26.9; CIDR4+7 = 31.5%). Pregnancy loss did not differ among treatments. Supplemental progesterone post-AI using a single intravaginal insert on d 4 was beneficial to pregnancy in cows inseminated following timed AI, but incremental progesterone with a second insert on d 7 did not improve fertility of dairy cows.     

Progesterone supplementation postinsemination improves fertility of cooled dairy cows during the summer

Reduced fertility of dairy cows during periods of elevated temperature, humidity, or both might be associated with low plasma progesterone concentration. Alleviation of thermal stress by efficient cooling is a prerequisite for improving fertility by hormonal treatment. We examined whether insertion of a controlled intravaginal drug-releasing (CIDR) insert containing progesterone following artificial insemination (AI) would improve summer conception rate. Control (n = 195) and treated (CIDR; n=165) cows, yielding on average 42.3 kg milk/d, were inseminated following estrus detection during the summer (July to October) in 2 commercial dairy herds in Israel. Mean maximal air temperature and relative humidity during the study were 30.2°C and 86%, respectively. All experimental cows were efficiently cooled throughout the study, as confirmed by measuring the body temperature of random cows. Treated cows received a CIDR insert on d 5 ± 1 post-AI for 13 d and pregnancy was confirmed by palpation 45 d post-AI. Plasma progesterone concentration in treated cows was elevated by approximately 1.5 ng/mL. Multiple logistic regressions were used to analyze conception rate. Treatment did not alter the overall conception rate; however, probability of conception increased in CIDR-treated cows with low body condition score (BCS) compared with their control counterparts (53 vs. 27%, respectively). A pronounced increase in probability of conception was recorded in CIDR-treated cows exhibiting both low BCS and postpartum reproductive disorders, compared with their control counterparts (58 vs. 14%, respectively). Exogenous progesterone supplementation on d 5 post-AI for 13 d improves summer fertility of subpopulations of cows exhibiting low BCS and postpartum reproductive disorders. Reproductive management based on specific hormonal treatment of designated subgroups of cows known to derive beneficial effects from it might improve treatment efficiency and reduce expenses.     

Induced lactation in nonpregnant cows: profitability and response to bovine somatotropin

Significant culling of high-producing cows with low fertility reduces profitability of dairy farms as those cows are replaced with heifers. Induced lactation of nonpregnant cows may be a management alternative to reduce culling and increase profits. The objectives of this study were to evaluate the efficacy of bovine somatotropin (bST) to increase milk production in cows induced into lactation with estrogen plus progesterone, and to determine the profitability of inducing cows into lactation vs. using replacement heifers entering the herd as first-lactation cows. Parity 1 or greater, nonpregnant, healthy Holstein cows (n = 28) were induced into lactation by administration of estradiol-17beta (0.075 mg/kg of body weight [BW] per d) and progesterone (0.25 mg/kg of BW per d) for 7 d. Milking began on d 18. Cows were randomly assigned to control or bST treatment groups on d 37 +/- 20 of milking, and milk production was compared for 70 d. After the 70-d comparison, all cows received bST for the duration of lactation. Cows receiving bST produced more milk (28.4 kg/d) than controls (24.1 kg/d), with variable yields among cows. For the economic analysis, induced cows were compared to first-lactation cows in the same herd using fair market value for costs and multiple component pricing for milk. Net present value for an induced cow (1966 dollars) was significantly greater than that for a first-lactation cow (1446 dollars). Our data suggest that bST use in induced cows is profitable. If a reliable method were developed and approved by the FDA, inducing nonpregnant cows into lactation could be used by dairy producers to increase profitability.     

Early postpartum reproductive profiles in Holstein cows with retained placenta and uterine discharges

Sixty Holstein cows were allocated to three groups. Twenty cows had retained placenta. The remaining cows were examined on d 14 postpartum and those with purulent discharges (n = 22) were assigned to one group and the remaining (n = 18) to a control group. Within each group, cows were given randomly either gonadotropin-releasing hormone (i.m., 200 micrograms) or saline on d 15 postpartum to evaluate the effect on changes in ovarian structures and plasma progesterone through 50 d postpartum and fertility. Corpora lutea were found in control cows by d 21, cows with uterine discharge by d 28, and cows with retained placenta by d 27. Maximum progesterone production during the first luteal phase was higher in control cows than in cows with purulent discharge or retained placenta (4.66 ng/ml compared with 3.23 and 3.34 ng/ml, respectively). Duration of the first corpus luteum was affected by clinical condition. Only 6.2% of cows with retained placenta had corpora lutea of normal duration (12 to 16 d), whereas 43.8 and 50.0% of cows with uterine discharge and control cows had normal postpartum luteal phases. Measures of fertility were not affected by gonadotropin-releasing hormone. Control cows had less days to conception (97) and fewer services per conception (1.6) than cows with retained placenta (134 and 2.5, respectively). Clinical group affected reproduction more than gonadotropin-releasing hormone did, possibly by altering ovarian function.     

Interactions of human chorionic gonadotropin with genotype and parity on fertility responses of lactating dairy cows

Fertility-promoting effects of treatment of lactating dairy cattle with human chorionic gonadotropin (hCG) after artificial insemination (AI) have been variable. Here, we tested whether fertility response to hCG in lactating Holstein cows interacts with genotype and parity. Primiparous (n = 538) and multiparous (n = 613) cows were treated with hCG (3,300 IU) or vehicle 5 d after AI. Pregnancy was diagnosed on d 32 and 60 after AI. A subset of cows (n = 593–701) was genotyped for 4 single nucleotide polymorphisms (SNP) previously associated with fertility. Treatment with hCG increased progesterone concentration on d 12 after AI regardless of genotype or parity. Pregnancy per AI was improved by hCG in primiparous cows but not in multiparous cows. Moreover, hCG treatment interacted with a SNP in coenzyme Q9 (COQ9) to affect fertility. Fertility of cows treated with vehicle was greatest for the AA allele, whereas fertility was lowest for the same genotype among cows treated with hCG. Pregnancy per AI was also affected by genotype for heat shock protein A1-like (HSPA1L) and progesterone receptor (PGR), but no interactions were observed with treatment. Genotype for a SNP in prostate androgen-regulated mucin-like protein 1 (PARM1) was not associated with fertility. Overall, results show that variation in response to hCG treatment on fertility depends on parity and interacts with a SNP in COQ9.     

Anogenital distance is associated with postpartum estrous activity,intensity of estrous expression,ovulation, and progesterone concentrations in lactating Holstein cows

The objectives of this retrospective observational study were to determine the associations of anogenital distance (AGD) with (a) postpartum estrous activity, (b) diameter of the preovulatory follicle, (c) intensity of estrous expression, (d) postestrus ovulation, (e) corpus luteum (CL) size, and (f) concentrations of progesterone at estrus and on d 7 after estrus. Lactating Holstein cows (n = 178; 55 primiparous, 123 multiparous) were enrolled into the study during the first postpartum week. All cows were continuously monitored by a pedometer-based automated activity monitoring (AAM) system for estrus. Postpartum estrous activity was assessed using the AAM estrus alerts, in which cows with at least one true estrus alert (i.e., a relative increase in steps from each cow's baseline detected by the AAM and the presence of at least one follicle >15 mm, a CL <20 mm, or no CL detected by ultrasound) by the first 50 d in milk (DIM) were considered to have commenced estrous activity. At the estrus alert >60 DIM, ovulation was determined by ultrasound at 24 h, 48 h, and 7 d after estrus, and blood samples were collected at estrus alert and on d 7 after estrus for progesterone analysis. The AGD was measured from the center of the anus to the base of the clitoris and classified as either short- or long-AGD using 2 cut-points of 148 mm (predictive of the probability of pregnancy to first insemination; short-AGD, n = 115; long-AGD, n = 63) and 142 mm (the median AGD; short-AGD, n = 90; long-AGD, n = 88). Regardless of the cut-point used, early postpartum estrous activity by 50 DIM (67 vs. 54%), duration of estrus (11.6 vs. 9.7 h), and preovulatory follicle diameter (20 vs. 19 mm) were greater in short-AGD than in long-AGD cows. Increased peak of activity at estrus in short-AGD cows (354 vs. 258% mean relative increase) was affected by an interaction between AGD and parity in which multiparous long-AGD cows had lesser relative increase in activity than primiparous cows (217 vs. 386%, respectively). Mean progesterone concentration at estrus was lesser in short-AGD (0.47 vs. 0.61 ng/mL) than in long-AGD cows. The ovulatory response at 24 h did not differ, but at 48 h (91 vs. 78%) and on d 7 after estrus (97 vs. 84%) it was greater in short-AGD cows. Although CL diameter on d 7 after estrus did not differ, short-AGD cows had greater progesterone concentration 7 d after estrus than long-AGD cows (4.1 vs. 3.2 ng/mL, respectively). In conclusion, greater proportions of short-AGD cows commenced estrous activity by 50 DIM, had larger preovulatory follicles, exhibited greater duration of estrus, had reduced progesterone concentration at estrus, had greater ovulation rates and progesterone concentration 7 d after estrus compared with long-AGD cows, with no difference in CL size between AGD groups. Because all the differences in physiological characteristics of short-AGD cows reported herein favor improved reproductive outcomes, we infer that these are factors contributing to improved fertility reported in short-AGD cows compared with long-AGD cows.     

Intensity of estrus following an estradiol-progesterone-based ovulation synchronization protocol influences fertility outcomes

The objective of this study was to examine the association between increased physical activity at the moment of timed artificial insemination (AI), detected by an automated activity monitor (AAM), and fertility outcomes. This paper also investigated factors affecting estrous expression in general. A total of 1,411 AI events from 1,040 lactating Holstein cows were recorded, averaging 1.3 ± 0.6 (±standard deviation) events per cow. Activity (measured as steps/h) was monitored continuously by a leg-mounted AAM located on the rear leg of the cow. Ovulation was synchronized by a timed AI protocol based on estradiol and progesterone. Ovarian ultrasonography was performed in all cows on d ?11 (AI = d 0) and in a subset of cows on d 0 (n = 588) and d 7 (n = 819) to determine the presence of a corpus luteum and follicles. The body condition score (1 to 5 scale) was assessed on d 0 and a blood sample was collected for progesterone measurement on d 7. Using the AAM, an estrus event was determined when the relative increase (RI) in physical activity of the cow exceeded 100% of the baseline activity. The physical activity was classified as strong RI (≥300% RI), moderate RI (100–300% RI), or no estrus (<100% RI). Milk production was measured daily and averaged between d ?11 and 0. Pregnancy was diagnosed at 32 and 60 d post-AI and pregnancy losses were calculated. The mean RI at estrus was 328.3 ± 132.1%. Cows with strong RI had greater pregnancy per AI than those with moderate RI and those that did not express estrus (35.1 vs. 27.3 vs. 6.2%). When including only cows that successfully ovulated after timed AI, those that displayed strong intensity RI still had greater pregnancy per AI than those with moderate intensity RI or those that did not express estrus (45.1 vs. 34.8 vs. 6.2%). Cows expressing strong RI at timed AI had greater ovulation rates compared with moderate RI and cows that did not express estrus (94.9 vs. 88.2 vs. 49.5%). Furthermore, pregnancy losses were reduced in cows with strong RI compared with cows expressing moderate RI (13.9 vs. 21.7%). Cows with a strong RI at estrus were more likely to have a corpus luteum at the beginning of the protocol and had greater concentration of progesterone 7 d post-AI. Multiparous cows expressed lower RI compared with primiparous cows. Cows with lower body condition score tended to have decreased RI at estrus. No correlation between estrous expression and pre-ovulatory follicle diameter was observed. Also, no correlation was observed between milk production at AI and RI. In conclusion, strong intensity RI of estrus events at timed AI was associated with improved ovulation rates and pregnancy per AI, and reduced pregnancy losses. These results provide further evidence that measurements of estrous expression can be used to predict fertility at the time of AI and possibly be used as a tool to assist decision making strategies of reproduction programs.