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The objective of this study was to evaluate four different automated activity monitoring (AAM) systems to identify anovulatory cows in early lactation. A total of 852 lactating Holstein cows (221 primiparous and 631 multiparous cows) from four commercial dairy herds were enrolled. On each farm, cows were equipped with a respective AAM system (SB: Smartbow; HT: Heatime; DP: Delpro; and CM: CowManager). Each cow was sampled three times within the voluntary waiting period (VWP) in a two-week interval to detect the blood progesterone (P4) concentration. Cows were classified based on the concentration of P4 as follows: (1) none of the three blood P4 concentrations exceeded 1.0 ng/mL (anovulatory); (2) at least one of the three blood P4 concentrations was above 1.0 ng/mL (ovulatory). Cows were classified based on estrus alerts as follows: (1) no estrus alert was detected by an AAM system from 7 to 60 DIM (anestrus); (2) at least one estrus alert was detected by an AAM system from 7 to 60 DIM (estrus). Accuracy, sensitivity, specificity, positive predictive value, and negative predictive value were calculated for each AAM system for anovulatory cows [(SB: 77.6%; 26.8%; 89.3%; 36.7%; and 84.1%); (HT: 79.2%; 63.6%; 83.6%; 52.8%; and 88.9%); (DP: 47.2%; 78.8%; 41.5%; 19.5%; and 91.6%); (CM: 80.5%; 23.7%; 92.7%; 39.1%; and 85.3%)].

Investigations on the optimum timing of artificial insemination (AI) following automated activity monitoring (AAM) depending on different types of semen in heifers are limited and in part show controversial results. Therefore, the objective of this observational study was to determine the association between the timing of AI using different characteristics of estrus (i.e., the onset, peak, and end of estrus) and pregnancy per AI (P/AI) in Holstein heifers. Heifers were fitted with a neck-mounted AAM system and inseminated with frozen conventional and sexed semen. The pregnancy per AI ( n = 4159) from 2858 heifers from six commercial dairy farms in Germany inseminated upon the alert of an AAM system was evaluated. Estrous intensity was classified based on peak activity into low (35 to 89 index value) and high (90 to 100 index value). We detected a quadratic association between the interval from the onset of estrus to AI and P/AI ( p = 0.02). The greatest P/AI was observed for heifers inseminated from 9 to 32 h after the onset of estrus. The intervals from the peak of activity to AI and the end of estrus to AI were not associated with P/AI ( p ≥ 0.05). Heifers inseminated with frozen conventional semen (50.1%) had a greater P/AI compared with heifers inseminated with frozen sexed semen (43.3%; p = 0.03). There were no interactions between the intervals from the onset, peak, or end of estrus to AI or the type of semen and the P/AI ( p ≥ 0.05). The pregnancy per AI was not associated with estrous intensity (50.5% for low intensity vs. 53.0% for high intensity; p = 0.37). In conclusion, inseminating heifers between 9 and 32 h after the onset of estrus, as detected by the AAM, optimized the P/AI regardless of semen type.

Dry cow treatment with an intramammary antibiotic is recommended to reduce the risk of mastitis at the beginning of the next lactation. The dry period may be shortened unintentionally, affecting antibiotic residue depletion and the time when residues reach concentrations below the maximum residue limit (MRL). The objective of this study was to evaluate residue depletion in milk after dry cow treatment with cloxacillin, considering dry periods of 14 (G14d), 21 (G21d), and 28 d (G28d). Overall, fifteen cows with 60 udder quarters were included in the study. For each cow, three of the udder quarters were treated with 1000 mg cloxacillin benzathine (2:1) on d 252, d 259, and d 266 of gestation; one quarter was left untreated. Milk samples were drawn until 20 DIM and milk composition, somatic cell count and cloxacillin residues were analyzed. The HPLC-MS/MS revealed different excretion kinetics for the compounds cloxacillin and cloxacillin benzathine (1:1). All cows showed a cloxacillin and cloxacillin benzathine (1:1) concentration below the MRL of 30 µg/kg after 5 d. In the udder quarters of G21d and G28d, the cloxacillin concentration was already below the MRL at first milking after calving. The cloxacillin benzathine (1:1) concentration in the milk of G28d, G21d, and G14d fell below 30 µg/kg on the 5th, 3rd, and 5th DIM, respectively. Shortening the dry period affects residue depletion after dry cow treatment with cloxacillin. The risk of exceeding the MRL, however, seems low, even with dry periods shorter than 14 d.

The objective of this observational study was to evaluate the association between increased physical activity at first artificial insemination (AI) and subsequent pregnancy per AI (P/AI) in lactating Holstein cows following spontaneous estrus or a timed AI (TAI) protocol. We also wanted to identify factors associated with the intensity of activity increase (PA) captured by automated activity monitors (AAM) and fertility. Two experiments were conducted, in which cows either were inseminated based on the alert of the AAM system (AAM cows) or received TAI following a 7-d Ovsynch protocol (TAI cows) if not inseminated within a farm-specific period after calving. Experiment 1 included 2,698 AI services from AAM cows and 1,042 AI services from TAI cows equipped with the Smarttag Neck (Nedap Livestock Management) from a dairy farm in Slovakia (farm 1). In the second experiment, 6,517 AI services from AAM cows and 1,226 AI services from TAI cows fitted with Heatime (Heatime Pro; SCR Engineers Ltd.) from 8 dairy farms in Germany (farms 2-9) were included. Pregnancy diagnosis was performed on a weekly basis by transrectal ultrasound (farms 1, 3, 7, 8) or by transrectal palpation (farms 2, 4-6, 9). Estrous intensity was represented by the peak value of the change in activity. In experiment 1, PA was categorized into low (x-factor 0-20) and high (x-factor 21-100) PA, and in experiment 2 into low (activity change = 35-89) and high (activity change = 90-100) PA. In TAI cows from both experiments, PA was additionally categorized into cows with no AAM alert. Data were analyzed separately for AAM and TAI cows using multinomial logistic regression models for PA in TAI cows and logistic regression models for PA in AAM cows and P/AI in both groups. In experiment 1, P/AI of AAM cows was greater for AI services performed with conventional frozen semen (57.6%) compared with sexed semen (47.2%), whereas type of semen only tended to be associated with P/AI in TAI cows (54.4% conventional frozen semen vs. 48.9% sexed semen). In experiment 2, P/AI was greater for fresh semen (AAM cows: 44.4% vs. TAI cows: 44.2%) compared with conventional frozen semen (AAM cows: 37.6% vs. TAI cows: 34.6%). In both experiments, pregnancy outcomes were associated with PA. In experiment 1, AAM cows with high PA (55.1%) had greater P/AI than cows with low PA (49.8%). Within TAI cows, cows with no alert (38.8%) had reduced P/AI compared with cows with low (54.2%) or high PA (61.8%). In experiment 2, AAM cows with high PA (45.8%) had greater P/AI compared with cows with low PA (36.4%). Timed AI cows with no alert (27.4%) had decreased P/AI compared with cows with low (41.1%) or high (50.8%) PA. The greatest risk factors for high PA were parity (experiment 1) and season of AI (except for TAI cows from experiment 1). We conclude that high PA at the time of AI is associated with greater odds of pregnancy for both AAM and TAI cows. In both experiments, about 2 thirds of AAM cows (experiment 1: 69.9% and experiment 2: 70.7%) reached high PA, whereas only approximately one-third or less of TAI cows (experiment 1: 37.3% and experiment 2: 23.6%) showed high PA. Although we observed similar results using 2 different AAM systems for the most part, risk factors for high PA might differ between farms and insemination type (i.e., AAM vs. TAI)., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this observational study was to evaluate the association of transition cow health and estrous expression, detected by an automated activity monitoring system (Smarttag Neck, Nedap Livestock Management), with reproductive performance in lactating Holstein cows. A total of 3,750 lactating Holstein cows (1,563 primiparous cows and 2,187 multiparous cows) from a commercial dairy farm in Slovakia calving from January 2020 until July 2021 were enrolled on an ongoing basis. Activity data were recorded from d 7 until d 60 postpartum. Within this observational period, cows were classified into 3 categories: (1) no estrus event (Estrus0), (2) 1 estrus event (Estrus1), or (3) 2 or more estrus events (Estrus2+). Transition cow health was assessed by farm personnel within the first 30 d in milk (DIM) using standard operating procedures. Generalized linear mixed models were used to analyze continuous and categorical data. Cox proportional hazard models were used for time to event data. The overall prevalence of anestrus was 20.8%. Multiparous cows had a greater risk for anestrus compared with primiparous cows [odds ratio (OR) = 1.4]. Cows with stillbirth (OR = 1.76), retained placenta (OR = 2.19), puerperal metritis (OR = 1.48), or subclinical ketosis (OR = 1.51) had a greater risk for anestrus. In addition, cows calving in summer (OR = 0.82), autumn (OR = 0.38), or winter (OR = 0.56) had a higher incidence of anestrus than cows calving in spring. Estrous expression from d 7 until d 60 postpartum was associated with estrous duration (DU) and estrous intensity at first artificial insemination (AI). Cows in Estrus0 had the shortest DU at first postpartum AI (9.4 ± 0.18 h) compared with cows in Estrus1 (10.5 ± 0.13 h) and Estrus2+ (11.4 ± 0.12 h). Cows in Estrus2+ had a longer DU at first postpartum AI compared with cows in Estrus1. For Estrus0, Estrus1, and Estrus2+ cows, pregnancy per AI at first service was 42.5%, 50.9%, and 55.4%, respectively. Estrous expression from d 7 until d 60 postpartum was associated with time to first AI and time to pregnancy. Compared with Estrus0 cows, Estrus1 [hazard ratio (HR) = 1.43] and Estrus2+ cows (HR = 1.62) had an increased hazard of being inseminated within 100 DIM. Compared with Estrus2+, Estrus1 cows had a reduced hazard of being inseminated within 100 DIM (HR = 0.89). Compared with Estrus0 cows, Estrus1 (HR = 1.24) and Estrus2+ cows (HR = 1.46) had an increased hazard of becoming pregnant within 200 DIM. Median DIM to pregnancy were 121, 96, and 92 for Estrus0, Estrus1, and Estrus2+ cows, respectively. In conclusion, cows with transition cow disorders (i.e., stillbirth, retained placenta, puerperal metritis, or subclinical ketosis) had a greater chance for anestrus compared with healthy cows. Cows in Estrus0 had reduced estrous expression at first AI and inferior reproductive performance compared with cows that displayed estrous activity from d 7 until d 60., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Incomplete luteal regression after treatment with a single dose of PGF 2α during an Ovsynch protocol decreases fertility to timed artificial insemination (AI). An additional treatment with PGF 2α 24 h after the first dose has been recommended to increase the proportion of cows with complete luteal regression and subsequent pregnancy per AI (P/AI). This is, however, associated with additional costs of labor and product. The objective was to develop a stochastic partial budget model to estimate the economic impact of an additional PGF 2α treatment on d 8 during an Ovsynch protocol in primiparous and multiparous cows. A systematic review of the literature and a meta-analytical assessment was performed to evaluate the effects of adding a second PGF 2α treatment during the Ovsynch protocol on P/AI in lactating dairy cows. Thirteen randomized controlled experiments from 11 published manuscripts including 9,735 cows were used. We were able to retrieve information regarding parity from 9 experiments from 7 manuscripts (2,367 primiparous cows and 5,356 multiparous cows). An additional dose of PGF 2α yielded a 5.60 risk difference in P/AI [95% confidence interval (CI) = 3.69-7.52]. In primiparous cows and multiparous cows, an additional dose of PGF 2α yielded a 4.24 (95% CI = 0.31-8.17) and a 5.31 risk difference in P/AI (95% CI = 2.75-7.87). Revenue was based on the associated improvement in reproductive performance because of an increase in P/AI multiplied by the value of a pregnancy (PGVAL). Median PGVAL was €252, ranging from €42 (fifth percentile) to €623 (95th percentile). Based on parity, median PGVAL was €205 (5th percentile = €43; 95th percentile = €651) and €264 (5th percentile = €88; 95th percentile = €598) for primiparous (n = 1,252) and multiparous cows (n = 3,003), respectively. Using a stochastic simulation model with 10,000 iterations, adding a second PGF 2α dose on d 8 was more profitable (€7.76/cow; 5th percentile = €0.01; 95th percentile = €29.40) compared with a single PGF 2α administration on d 7. In primiparous cows, adding a second PGF 2α treatment was more profitable (€1.99/cow; 5th percentile = -€3.08; 95th percentile = €22.52) in 67% of all simulated iterations. In multiparous cows, adding a second PGF 2α treatment was more profitable (€7.92/cow; 5th percentile = -€0.09; 95th percentile = €28.22) in 95% of all simulated iterations. In conclusion, there was a clear benefit of an additional PGF 2α treatment during the Ovsynch protocol on P/AI (+5.6 percentage units). Despite additional costs for hormones and labor, an additional treatment with PGF 2α on d 8 was more profitable in 95% of all scenarios because of the associated increase in fertility. It turned out, however, that adding a second PGF 2α treatment was more profitable in multiparous cows compared with primiparous cows., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this observational study was to evaluate the association of estrous expression within 40 days in milk (DIM) using a neck-mounted automated activity monitor (Heatime Pro; SCR Engineers Ltd.) with reproductive performance in lactating Holstein cows. A total of 2,077 cows (614 primiparous cows and 1,463 multiparous cows) from 5 commercial dairy farms were included in the statistical analyses. Activity data from the first 7 d after calving were excluded. An estrus event was defined as an activity change index ≥35 for more than 2 h. Cows were classified according to the number of estrus events from d 7 until d 40 postpartum into 3 categories: (1) no estrus event (Estrus0); (2) one estrus event (Estrus1), and (3) 2 or more estrus events (Estrus2). Generalized linear mixed models were used to analyze continuous and categorical data. Shared frailty models were used for time to event data. Overall, 52.7% of cows had no estrus event detected by an automated activity monitor system from d 7 until d 40 postpartum. Herd level prevalence of Estrus0 ranged from 37.5 to 58.4%. Estrous expression from d 7 until d 40 postpartum affected estrous duration and estrous intensity at first artificial insemination (AI). Cows in Estrus0 had the shortest duration (13.2 ± 0.33 h) compared with cows in Estrus1 (13.8 ± 0.36 h) and Estrus2 (14.8 ± 0.41 h). Cows in Estrus2 had a longer estrous duration at first postpartum AI compared with cows in Estrus1. Among Estrus0 cows, 46.2% had an estrus event with high intensity at first postpartum AI. Among cows in Estrus1 and Estrus2, 50.8 and 53.8% had an estrus event with high intensity at first postpartum AI, respectively. There was a significant difference between Estrus2 and Estrus0 and a tendency between Estrus0 and Estrus1. There was no difference between Estrus1 and Estrus2. For Estrus0, Estrus1, and Estrus2 cows, pregnancy per AI was 29.4, 30.9, and 37.8%, respectively. There was a significant difference between Estrus0 and Estrus2 and Estrus1 and Estrus2. There was no difference between Estrus0 and Estrus1. Estrous expression from d 7 until d 40 postpartum affected time to first AI and time to pregnancy. Compared with Estrus0 cows, cows in Estrus1 [hazard risk (HR) = 1.74] and Estrus2 (HR = 1.77) had an increased hazard of being inseminated within 100 DIM. There was no difference between Estrus1 and Estrus2. Median DIM to first AI were 70, 59, and 58 for cows in Estrus0, Estrus1, and Estrus2, respectively. Compared with Estrus0 cows, cows in Estrus1 (HR = 1.28) and Estrus2 (HR = 1.33) had an increased hazard of becoming pregnant within 200 DIM. There was no difference between Estrus1 and Estrus2. Median DIM to pregnancy were 127, 112, and 103 for Estrus0 cows, Estrus1 and Estrus2, respectively. In conclusion, cows with no estrous expression from 7 to 40 DIM had reduced estrous expression at first AI and inferior reproductive performance compared with cows that displayed estrous activity., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this observational study was to identify factors associated with estrous duration (DU) and intensity measured as the peak of activity (PA) change and subsequent fertility in lactating Holstein cows using a neck-mounted automated activity monitor (Heatime Pro, SCR Engineers Ltd., Netanya, Israel). Ambient temperature and relative humidity were recorded hourly to calculate the temperature-humidity index (THI). A total of 5,933 estrus events from 3,132 cows located on 8 commercial dairy farms in Germany were used for this study. Farms participated in monthly DHIA testing. Pregnancy diagnosis was performed either by transrectal palpation [farm 1: 42 ± 3 d; farm 3: 40 ± 3 d; farms 4 and 8: 38 ± 3 d; farm 5: 43 ± 3 d after artificial insemination (AI)] or transrectal ultrasonography (farms 2, 6, and 7: 30 ± 3 d after AI). Estrous intensity was categorized based on peak activity of estrus into low (35-89 index value), and high (90-100 index value) PA. Overall, 73.5% of estrus events were of high PA. The mean (± standard error of the mean) DU was 14.94 ± 0.06 h. There was a strong correlation between DU and PA (r = 0.67). In the final statistical model, only PA was associated with pregnancy per artificial insemination (P/AI), with 1.35 greater odds of pregnancy for cows with high PA compared with cows with low PA. Increased THI 1 wk before AI was associated with shorter DU, lower PA, and decreased P/AI. A small percentage of cows (4.7%) showed short interestrus intervals (i.e., more than 1 activity peak within 7 d close to the event of estrus), resulting in reduced DU, PA, and P/AI. The change of weighted rumination was associated with DU and PA, as a lower nadir was associated with a greater risk for high PA and long DU. There was no association, however, between the nadir of change of weighted rumination and P/AI. Whereas milk yield and somatic cell count from the DHIA test date before AI were negatively associated with estrous expression, neither milk yield nor somatic cell count was associated with P/AI. Surprisingly, multiparous cows expressed estrus with longer DU (13.15 ± 0.31 h) compared with primiparous cows (12.52 ± 0.32 h), whereas PA did not differ among parities. Pregnancy per AI was greater for primiparous (29.4%) than for multiparous (22.1%) cows. An estrus event with long DU or high PA was more likely later in lactation. Milk fat, milk protein, milk urea nitrogen, and lactose from the DHIA test date closest to AI had no association with estrous expression or P/AI. In conclusion, DU and PA were highly correlated, and cows with high PA were particularly associated with greater odds for pregnancy. A negative association between estrous expression and P/AI was identified for increased THI 1 wk before AI and cows with short interestrus intervals using automated activity monitor., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to evaluate the effect of two prostaglandin F 2α (PGF) treatments 24 h apart (500 μg of cloprostenol) and treatment with a double PGF dose on d 7 (1000 μg of cloprostenol) during a 7-d Ovsynch protocol on progesterone (P4) concentration and pregnancy per artificial insemination (P/AI) in lactating Holstein cows. We hypothesized that treatment leads to a decreased P4 concentration at the second GnRH treatment (G2) and an increase in P/AI compared to the traditional 7-d Ovsynch protocol. A secondary hypothesis was that the treatment effect is influenced by the presence of a corpus luteum (CL) at the first GnRH treatment (G1). Two experiments were conducted on 8 commercial dairy farms in Germany. Once a week, cows from both experiments were assigned in a consecutive manner to receive: (1) Ovsynch (control: GnRH; 7 d, PGF; 9 d, GnRH), (2) Ovsynch with a double PGF dose (GDPG: GnRH; 7 d, 2xPGF; 9 d, GnRH), or (3) Ovsynch with a second PGF treatment 24 h later (GPPG: GnRH; 7 d, PGF; 8 d, PGF; 32 h, GnRH). All cows received timed AI (TAI) approximately 16 h after G2. Pregnancy diagnosis was performed by transrectal palpation (38 ± 3 d after TAI, experiment 1) or transrectal ultrasonography (35 ± 7 d after TAI, experiment 2). Whereas farms from experiment 1 used a Presynch-Ovsynch protocol (PGF, 14 d later PGF, 12 d later GnRH, 7 d later PGF, 2 d later GnRH, and 16-18 h later TAI) to facilitate first postpartum TAI, no presynchronization protocol was used on farms from experiment 2. In experiment 1, we enrolled 1581 lactating dairy cows (60 experimental units) from 2 dairy farms. At G2, blood samples were collected from a subsample of cows (n = 491; 16 experimental units) to determine P4 concentration at G2. In experiment 2, we enrolled 1979 lactating dairy cows (252 experimental units) from 6 dairy farms. Transrectal ultrasonography was performed to determine the presence or absence of a CL at G1. In experiment 1, treatment affected P/AI (P = 0.01) and P/AI was greater for GDPG (38.2%) and GPPG (38.9%) than for control cows (29.8%). Both, GDPG and GPPG cows had decreased P4 concentration at G2 compared with control cows (P < 0.01). Whereas both treatments increased the percentage of cows with very low P4 concentration (0.00-0.09 ng/mL) at G2, only the GPPG treatment decreased the percentage of cows with high P4 concentration (≥0.6 ng/mL) at G2 compared to the control group. In experiment 2, P/AI was greater for GPPG (37.4%) than for control cows (31.0%; P = 0.03) and tended to be greater than for GDPG cows (31.8%; P = 0.05). Cows from the GDPG group had similar (P = 0.77) P/AI compared to the control group. Pregnancy per AI did not differ between cows with a CL at G1 and cows without a CL at G1 (34.1% vs. 32.6%; P = 0.50). There was no interaction between treatment and presence of a CL at G1 on P/AI (P = 0.61). Combining data from the 2 experiments but excluding cows from experiment 1 receiving presynchronization before first TAI (n = 2573; 312 experimental units), P/AI was greater for GPPG (40.3%; P < 0.01) than for control (31.8%) and GDPG cows (33.4%). Between GDPG and control cows, P/AI did not differ (P = 0.46). We conclude that overall the addition of a second PGF treatment on d 8 during a 7-d Ovsynch protocol increased P/AI compared to the traditional 7-d Ovsynch including a single PGF dose on d 7 and to a double PGF dose on d 7. Doubling the PGF dose on d 7 in a 7-d Ovsynch protocol did not affect P/AI. Use of a presynchronization protocol, however, seems to influence the effect of a dose frequency modification of PGF treatment in an Ovsynch protocol. Presynchronized cows receiving first postpartum TAI had similarly increased P/AI treated with a double PGF dose compared with treatment with a second PGF dose. Future studies need to elucidate whether the treatment effect is modified by presynchronization of the first postpartum TAI., Competing Interests: Declaration of competing interest Neither of the authors of this paper has a financial or personal relationship with other people or organisations that could inappropriately influence or bias the content of the paper., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The objective of this observational experiment was to determine the association between the time of artificial insemination (AI) and pregnancy per AI (P/AI) in lactating Holstein cows inseminated with either fresh or frozen semen considering different characteristics of an estrus event (i.e., onset, peak, and end) using an automated activity monitoring system. A total of 3,607 AI services based on the alert of an automated activity monitoring system (Heatime; SCR Engineers Ltd., Netanya, Israel) were evaluated from 4 commercial dairy farms in Germany. Pregnancy diagnosis was performed by transrectal palpation 38 ± 3 d after AI or by transrectal ultrasonography 30 ± 3 d after AI. Estrus intensity was categorized based on peak activity of estrus (PAE) into low (35-89 index value) and high (90-100 index value) intensity. The mean (± standard deviation) duration of an estrus event was 14.3 ± 4.6 h. The mean (± standard deviation) interval from onset of estrus (OE; moment where index value was ≥35) to AI was 16.8 ± 8.0 h, from PAE to AI was 11.9 ± 8.1 h, and from end of estrus (EE; moment where index value returned to <35) to AI was 2.5 ± 8.7 h. Primiparous cows had greater P/AI than multiparous cows, whereas first AI postpartum yielded greater P/AI compared with subsequent AI services. Type of semen was not associated with P/AI. Cows with heat stress 1 wk before AI had decreased P/AI. Cows with low estrus intensity (26.0%) were less fertile compared with cows showing high estrus intensity (32.8%). Cows with intermediate 100-d milk yield had decreased P/AI compared with cows with either low or high 100-d milk yield. There was a quadratic effect of the interval from OE to AI on P/AI. At 38 d after AI, P/AI was greatest for cows inseminated from 7 to 24 h after OE, within 18 h after PAE, or from 5 h before EE to 12 h after EE. There was no interaction between the interval from OE to AI and type of semen. There tended to be an interaction between the intervals from PAE to AI and type of semen and from EE to AI and type of semen. Cows inseminated with fresh semen within 5 h before EE had greater P/AI compared with frozen semen, whereas cows inseminated with frozen semen from 13 to 18 h after EE had greater P/AI compared with fresh semen. In conclusion, inseminating cows from 7 to 24 h after OE or 1 to 18 h after PAE yielded greatest P/AI irrespective of type of semen. In addition, high estrus intensity was positively associated with P/AI., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)